identifier
stringlengths 11
32
| pdf_url
stringlengths 17
4.62k
⌀ | lang
stringclasses 120
values | error
stringclasses 1
value | title
stringlengths 2
500
⌀ | source_name
stringlengths 1
435
⌀ | publication_year
float64 1.9k
2.02k
| license
stringclasses 3
values | word_count
int64 0
1.64M
| text
stringlengths 1
9.75M
|
|---|---|---|---|---|---|---|---|---|---|
https://openalex.org/W2983014931
|
https://www.mdpi.com/2077-0383/8/11/1899/pdf?version=1573115060
|
English
| null |
Urinary Biomarkers α-GST and π-GST for Evaluation and Monitoring in Living and Deceased Donor Kidney Grafts
|
Journal of clinical medicine
| 2,019
|
cc-by
| 8,165
|
Received: 22 October 2019; Accepted: 5 November 2019; Published: 7 November 2019 Abstract: The aim of this study was to analyze the value of urine α- and π-GST in monitoring
and predicting kidney graft function following transplantation. In addition, urine samples from
corresponding organ donors was analyzed and compared with graft function after organ donation from
brain-dead and living donors. Urine samples from brain-dead (n = 30) and living related (n = 50) donors
and their corresponding recipients were analyzed before and after kidney transplantation. Urine α-
and π-GST values were measured. Kidney recipients were grouped into patients with acute graft
rejection (AGR), calcineurin inhibitor toxicity (CNI), and delayed graft function (DGF), and compared
to those with unimpaired graft function. Urinary π-GST revealed significant differences in deceased
kidney donor recipients with episodes of AGR or DGF at day one after transplantation (p = 0.0023 and
p = 0.036, respectively). High π-GST values at postoperative day 1 (cutoff: >21.4 ng/mg urine creatinine
(uCrea) or >18.3 ng/mg uCrea for AGR or DGF, respectively) distinguished between rejection and
no rejection (sensitivity, 100%; specificity, 66.6%) as well as between DGF and normal-functioning
grafts (sensitivity, 100%; specificity, 62.6%). In living donor recipients, urine levels of α- and π-GST
were about 10 times lower than in deceased donor recipients. In deceased donors with impaired
graft function in corresponding recipients, urinary α- and π-GST were elevated. α-GST values
>33.97 ng/mg uCrea were indicative of AGR with a sensitivity and specificity of 77.7% and 100%,
respectively. In deceased donor kidney transplantation, evaluation of urinary α- and π-GST seems to
predict different events that deteriorate graft function. To elucidate the potential advantages of such
biomarkers, further analysis is warranted. Keywords: kidney transplantation; urinary biomarkers; α-GST; π-GST; acute rejection; delayed graft
function; nephrotoxicity Journal of
Clinical Medicine Journal of
Clinical Medicine Journal of
Clinical Medicine Journal of
Clinical Medicine Urinary Biomarkers α-GST and π-GST for Evalu
and Monitoring in Living and Deceased Donor
Kidney Grafts Shadi Katou 1,*
, Brigitta Globke 2, M. Haluk Morgul 1, Thomas Vogel 1
,
Benjamin Struecker 1
, Natalie Maureen Otto 3, Anja Reutzel-Selke 2, Marion Marksteiner 2,
Jens G. Brockmann 1, Andreas Pascher 1 and Volker Schmitz 1 1
Department of General, Visceral and Transplantation Surgery, Universitätsklinikum Münster,
48189 Münter, Germany; haluk.morguel@ukmuenster.de (M.H.M.); thomas.vogel@ukmuenster.de (T.V.);
benjamin.struecker@ukmuenster.de (B.S.); jens.brockmann@ukmuenster.de (J.G.B.);
andreas.pascher@ukmuesnter.de (A.P.); schmitz.volker@icloud.com (V.S.) 1
Department of General, Visceral and Transplantation Surgery, Universitätsklinikum Münster,
48189 Münter, Germany; haluk.morguel@ukmuenster.de (M.H.M.); thomas.vogel@ukmuenster.de (T.V.);
benjamin.struecker@ukmuenster.de (B.S.); jens.brockmann@ukmuenster.de (J.G.B.);
andreas.pascher@ukmuesnter.de (A.P.); schmitz.volker@icloud.com (V.S.) p
(
)
(
)
2
Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität
zu Berlin, and Berlin Institute of Health, Department of Surgery CCM|CVK, 10117 Berlin, Germany;
brigitta.globke@charite.de (B.G.); anja.selke@charite.de (A.R.-S.); marion.marksteiner@charite.de (M.M.) p
2
Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität
zu Berlin, and Berlin Institute of Health, Department of Surgery CCM|CVK, 10117 Berlin, Germany;
brigitta.globke@charite.de (B.G.); anja.selke@charite.de (A.R.-S.); marion.marksteiner@charite.de (M.M.)
3
Department of Nephrology and Internal Intensive Care Medicine, Charité Universitätsmedizin Berlin, p
2
Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität
zu Berlin, and Berlin Institute of Health, Department of Surgery CCM|CVK, 10117 Berlin, Germany;
brigitta.globke@charite.de (B.G.); anja.selke@charite.de (A.R.-S.); marion.marksteiner@charite.de (M.M.)
3
Department of Nephrology and Internal Intensive Care Medicine, Charité Universitätsmedizin Berlin,
Campus Virchow Klinikum, 10117 Berlin, Germany; natalie.otto@charite.de
*
Correspondence: shadi.katou@ukmuenster.de; Tel.: +49-251-8351759 brigitta.globke@charite.de (B.G.); anja.selke@charite.de (A.R.-S.); marion.marksteiner@charite.de (M.M.)
3
Department of Nephrology and Internal Intensive Care Medicine, Charité Universitätsmedizin Berlin,
Campus Virchow Klinikum, 10117 Berlin, Germany; natalie.otto@charite.de
*
Correspondence: shadi.katou@ukmuenster.de; Tel.: +49-251-8351759 g
g
j
3
Department of Nephrology and Internal Intensive Care Medicine, Charité Universitätsmedizin Berlin,
Campus Virchow Klinikum, 10117 Berlin, Germany; natalie.otto@charite.de Campus Virchow Klinikum, 10117 Berlin, Germany; natalie.otto@charite.de
*
Correspondence: shadi.katou@ukmuenster.de; Tel.: +49-251-8351759 *
Correspondence: shadi.katou@ukmuenster.de; Tel.: +49-251-8351759 1. Introduction Kidney transplantation is by far the best therapeutic option for patients with end-stage renal
disease (ESRD). After transplantation, the main challenges, besides surgical complications, are acute J. Clin. Med. 2019, 8, 1899; doi:10.3390/jcm8111899 2 of 11 J. Clin. Med. 2019, 8, 1899 graft rejection, delayed graft function, and adverse effects of immunosuppressants [1]. Acute graft
rejection still occurs in up to 25% of recipients and is a significant prognostic factor for long-term
graft survival [2]. The improvements of immunosuppressive drugs have turned transplantation into a
safe and widely predictable therapy; however, many of the agents used today still contribute to graft
failure due to their nephrotoxic potential [3]. Delayed graft function, defined by the need for dialysis
within the first week after transplantation and mainly caused by acute tubular necrosis, is mostly
due to long ischemia times, advanced donor age, and comorbidities [4,5]. Recognizing the cause of
graft dysfunction may be challenging, yet immediate diagnosis and therapy are essential for optimal
graft survival. g
The signs of graft dysfunction are decreased diuresis and impaired creatinine blood levels. Monitoring immunosuppressive drugs and their toxicity through serum levels is of limited value,
since the difference between therapeutic and toxic levels is not fixed [6]. In order to define the
pathomechanism of graft dysfunction, a graft biopsy is required in most cases. However, this is an
invasive procedure and risks associated complications endangering the transplanted kidney [7]. α- and π-GST, which are specifically present in the kidney tubules, are two isotypes of the
glutathione-S-transferases. Beyond their biochemical differences, they are also located in different parts
of the tubule system [8]. α-GST is found in cells of the proximal tubules, which are predominantly
affected by ischemia time and nephrotoxic substances. π-GST is located in distal tubules, which are
damaged during acute graft rejection [9]. Their release into the urine as a result of cell damage
gives an accurate prediction of the impaired part of the tubules system and therefore the underlying
cause of graft dysfunction [10,11]. Analyses of α- and π-GST have been reported to be promising for
discriminating between the different causes of graft dysfunction [11–13]. The aim of our study was to determine the value of measuring α- and π-GST concentrations in
urine as biomarkers for monitoring graft function and predicting postoperative events in the first week
after transplantation in living and deceased donor kidney transplantation. 2.1. Samples and Data Collection This study was approved by the Ethics Committee of Berlin’s Charité University Hospital
(EA2/137/10). We prospectively analyzed blood and urine samples as well as demographic data from
160 patients: 30 brain-dead donors and their 30 corresponding recipients; and 50 living kidney donors
as well as their 50 corresponding living donor kidney recipients. All surgeries were carried out at the
Department of General, Visceral and Transplantation surgery of Charité University Hospital, Berlin. Machine perfusion was not performed for any kidney allograft in this study. Except for brain-dead
donors, all patients were followed during the first week after surgery. Blood and urine samples were
collected at the following time points: day 0, day 1, day 3, day 5, and day 7. Samples from brain-dead
organ donors were obtained on day 0, the day of the organ donation surgery. Urine samples were
collected from recipients after transplantation through an externalized uretero-vesico-cutaneous stent
and therefore exclusively reflected the α- and π-GST content of the transplanted grafts. 2 3 GST Analysis and Statistics
2.3. GST Analysis and Statistics 2 3 GST Analysis and Statistics
2.3. GST Analysis and Statistics y
Urinary α- and π-GST values were measured using a commercially available ELISA test kit
provided by Argutus Medical Ltd. (Dublin, Ireland). To consider the physiological differences in
urine concentrations, α- and π-GST values were standardized to urine creatinine. The resulting unit
for GST was ng/mg uCrea. Reference ranges of urinary α- and π-GST were determined as
recommended from the measured GST values in the healthy population of this study group. For
this purpose, the α- and π-GST values in the urine of healthy living donors were considered before
donor nephrectomy. The reference interval for α-GST is 2.7–7.6 ng/mg uCrea and for π-GST 4.1–13
ng/mg uCrea in this study. For statistical analysis, GraphPad Prism 6 (GraphPad Software, San Diego, CA, USA) was
used. Quantitative data are given as the mean and standard deviation. To compare normally
distributed variables, t-tests such as Mann‒Whitney and Wilcoxon were performed. For the
comparison of multiple variables, we used two-way ANOVA; here we applied Tukey and Holm‒
Sidak tests for post hoc analysis of the subgroups. The area under the curve was calculated in ROC
Urinary α- and π-GST values were measured using a commercially available ELISA test kit
provided by Argutus Medical Ltd. (Dublin, Ireland). To consider the physiological differences in urine
concentrations, α- and π-GST values were standardized to urine creatinine. The resulting unit for
GST was ng/mg uCrea. Reference ranges of urinary α- and π-GST were determined as recommended
from the measured GST values in the healthy population of this study group. For this purpose, the α-
and π-GST values in the urine of healthy living donors were considered before donor nephrectomy. The reference interval for α-GST is 2.7–7.6 ng/mg uCrea and for π-GST 4.1–13 ng/mg uCrea in this study. For statistical analysis, GraphPad Prism 6 (GraphPad Software, San Diego, CA, USA) was used. Quantitative data are given as the mean and standard deviation. To compare normally distributed
variables, t-tests such as Mann-Whitney and Wilcoxon were performed. For the comparison of multiple
variables, we used two-way ANOVA; here we applied Tukey and Holm-Sidak tests for post hoc analysis
of the subgroups. The area under the curve was calculated in ROC analysis and a log-rank (Mantel-Cox)
test was performed in survival analysis. A p-value less than 0.05 was defined as significant. 2 3 GST Analysis and Statistics
2.3. GST Analysis and Statistics y
Urinary α- and π-GST values were measured using a commercially available ELISA test kit
provided by Argutus Medical Ltd. (Dublin, Ireland). To consider the physiological differences in
urine concentrations, α- and π-GST values were standardized to urine creatinine. The resulting unit
for GST was ng/mg uCrea. Reference ranges of urinary α- and π-GST were determined as
recommended from the measured GST values in the healthy population of this study group. For
this purpose, the α- and π-GST values in the urine of healthy living donors were considered before
Urinary α- and π-GST values were measured using a commercially available ELISA test kit
provided by Argutus Medical Ltd. (Dublin, Ireland). To consider the physiological differences in urine
concentrations, α- and π-GST values were standardized to urine creatinine. The resulting unit for
GST was ng/mg uCrea. Reference ranges of urinary α- and π-GST were determined as recommended
from the measured GST values in the healthy population of this study group. For this purpose, the α-
and π-GST values in the urine of healthy living donors were considered before donor nephrectomy. Th
f
i t
l f
GST i 2 7 7 6
/
C
d f
GST 4 1 13
/
C
i thi
t d donor nephrectomy. The reference interval for α-GST is 2.7–7.6 ng/mg uCrea and for π-GST 4.1–13
ng/mg uCrea in this study. For statistical analysis, GraphPad Prism 6 (GraphPad Software, San Diego, CA, USA) was
used. Quantitative data are given as the mean and standard deviation. To compare normally
distributed variables, t-tests such as Mann‒Whitney and Wilcoxon were performed. For the
comparison of multiple variables, we used two-way ANOVA; here we applied Tukey and Holm‒
Sidak tests for post hoc analysis of the subgroups. The area under the curve was calculated in ROC
For statistical analysis, GraphPad Prism 6 (GraphPad Software, San Diego, CA, USA) was used. Quantitative data are given as the mean and standard deviation. To compare normally distributed
variables, t-tests such as Mann-Whitney and Wilcoxon were performed. For the comparison of multiple
variables, we used two-way ANOVA; here we applied Tukey and Holm-Sidak tests for post hoc analysis
of the subgroups. The area under the curve was calculated in ROC analysis and a log-rank (Mantel-Cox)
test was performed in survival analysis. A p-value less than 0.05 was defined as significant. 2.2. Immunosuppression Events and Subgroups All recipients received a triple immunosuppressant consisting of prednisolone, mycophenolate
mofetil (MMF), and a calcineurin inhibitor. All deceased donor kidney recipients received tacrolimus,
whereas living donor kidney recipients were treated with either tacrolimus or cyclosporine. Recipients
were divided into subgroups according to the events in the first postoperative week: acute graft
rejection (AGR, G1), calcineurin-induced nephrotoxicity (CNI, G2), both acute kidney rejection and
calcineurin-induced nephrotoxicity (AGR + CNI, G3), delayed graft function (DGF, G4), and event-free
(healthy, G5) subgroup (Figure 1). An acute graft rejection was confirmed by graft biopsy and classified
according to the BANFF criteria. Calcineurin-induced nephrotoxicity was defined by serum levels of 3 of 11
d graft J. Clin. Med. 2019, 8, 1899
both acute kidney rej agents (tacrolimus >15 ng/mL, cyclosporin >250 ng/mL). It is worth noting that delayed graft function,
characterized by the need for dialysis in the first week after transplantation, was not recorded in any of
the living donor recipients. In case dialysis was required due to a known graft-damaging event such
as AGR or CNI, those patients were enrolled in subgroups of primary cause and not defined as DGF. Recipients with no signs for any of the above events were considered healthy and were used as our
control group. confirmed by graft biopsy and classified according to the BANFF criteria. Calcineurin-induced
nephrotoxicity was defined by serum levels of agents (tacrolimus >15 ng/mL, cyclosporin >250
ng/mL). It is worth noting that delayed graft function, characterized by the need for dialysis in the
first week after transplantation, was not recorded in any of the living donor recipients. In case
dialysis was required due to a known graft-damaging event such as AGR or CNI, those patients
were enrolled in subgroups of primary cause and not defined as DGF. Recipients with no signs for
any of the above events were considered healthy and were used as our control group Figure 1. Patients, groups, and subgroups. Figure 1. Patients, groups, and subgroups. Figure 1. Patients, groups, and subgroups. Figure 1. Patients, groups, and subgroups. g
3 R
lt
3.1. Demographic Data 3. Results
3.1. Demographic Data
There were no differences regarding gender and BMI between the patient groups. The mean
eGFR before graft recovery or transplantation (d0) did not differ between the groups. Recipients of
deceased donor kidneys were significantly older than those of living donors (58 ± 13 and 48 ± 15
respectively, p = 0.006), whereas the donor’s age was not different. Cold as well as warm ischemic
time were significantly higher in the deceased donor group (both p < 0 0001) The demographic data
There were no differences regarding gender and BMI between the patient groups. The mean
eGFR before graft recovery or transplantation (d0) did not differ between the groups. Recipients of
deceased donor kidneys were significantly older than those of living donors (58 ± 13 and 48 ± 15
respectively, p = 0.006), whereas the donor’s age was not different. Cold as well as warm ischemic time
were significantly higher in the deceased donor group (both p < 0.0001). The demographic data of
donors and recipients are given in Table 1. The increase in the eGFR of recipients after transplantation
was, as expected, more noticeable in living donation scenarios (Figure 2). 4 of 11
p J. Clin. Med. 2019, 8, 1899 Table 1. Demographic data of donors and recipients. 58 ± 15
11/19 Deceased Donor Grafts
Living Donor Grafts
p Value
Donor
30
50
Age (years)
58 ± 15
53 ± 10
n.s. Sex (male/female)
11/19
22/28
n.s. BMI
27.3 ± 6.5
25.3 ± 3.2
n.s. eGFR, d0 (mL/min)
96 ± 39
97 ± 18
n.s. Diuresis in last hour (mL)
152 ± 81
-
Recipient
30
50
Age (years)
58 ± 12
47 ± 15
<0.05
Sex (male/female)
22/8
35/15
n.s. BMI
26.4 ± 4
25.8 ± 4.9
n.s. eGFR, d0 (mL/min)
9 ± 3
8 ± 4
n.s. Primary disease
Glomerulonephritis
9
24
n.s. Hypertensive nephrosclerosis
7
9
n.s. Polycystic kidney disease
6
2
n.s. Autoimmune disease
2
5
n.s. Diabetes
2
3
n.s. Urologic disease
2
2
n.s. Calcineurin-induced nephrotoxicity
0
2
n.s. Others
2
3
n.s. Dialysis
Hemodialysis
29
28
n.s. Peritoneal dialysis
1
3
n.s. No dialysis
0
19
<0.05
Cold ischemia time (minutes)
613 ± 269
193 ± 62
<0.05
Warm ischemia time (minutes)
34 ± 11
23 ± 7
<0.05
BMI, body mass index; eGFR, estimated glomerular filtration rate; n.s., not significant. g
3 R
lt
3.1. Demographic Data Sex (male/female)
11/19
22/28
BMI
27.3 ± 6.5
25.3 ± 3.2
eGFR, d0 (mL/min)
96 ± 39
97 ± 18
Diuresis in last hour (mL)
152 ± 81
-
Recipient
30
50
Age (years)
58 ± 12
47 ± 15
Sex (male/female)
22/8
35/15
BMI
26.4 ± 4
25.8 ± 4.9
eGFR, d0 (mL/min)
9 ± 3
8 ± 4
Primary disease
Glomerulonephritis
9
24
pertensive nephrosclerosis
7
9
olycystic kidney disease
6
2
Autoimmune disease
2
5
Diabetes
2
3
Urologic disease
2
2
eurin-induced nephrotoxicity
0
2
Others
2
3
Dialysis
Hemodialysis
29
28
Peritoneal dialysis
1
3
No dialysis
0
19
d ischemia time (minutes)
613 ± 269
193 ± 62
rm ischemia time (minutes)
34 ± 11
23 ± 7
BMI, body mass index; eGFR, estimated glomerular filtration rate; n.s., not significant. Deceased Donor Grafts
Living Donor Grafts
p Value
11/19
22/28
27.3 ± 6.5
25.3 ± 3.2 BMI, body mass index; eGFR, estimated glomerular filtration rate; n.s., not significant. y mass index; eGFR, estimated glomerular filtration rate; n.s., not s e 2. eGFR comparison between deceased and living donor recipients before and durin
Figure 2. eGFR comparison between deceased and living donor recipients before and during the first
week after transplantation. 2. eGFR comparison between deceased and living donor recipients before and duri
Figure 2. eGFR comparison between deceased and living donor recipients before and during the first
week after transplantation. t week after transplantation.
3.2. α- and π-GST in Recipients nd π-GST in Recipients
ither α- nor π-GST correlated with age, BMI, and cold or warm ischemic time in any
er, both GST isoenzymes correlated with renal function in living donors and in the h
ts subgroup. In deceased donor recipients and living donor recipients we observed
n in four (13.4%) and eight patients (16%), calcineurin-induced nephrotoxicity in five (
24%) patients, and both simultaneously in three (10%) and three (6%) patients, respec
graft function occurred only with deceased donors (nine patients, 30%). Patients w
ful postoperative course in the deceased donor recipient group numbered nine (30%
Neither α- nor π-GST correlated with age, BMI, and cold or warm ischemic time in any group. However, both GST isoenzymes correlated with renal function in living donors and in the healthy
recipients subgroup. In deceased donor recipients and living donor recipients we observed acute
rejection in four (13.4%) and eight patients (16%), calcineurin-induced nephrotoxicity in five (16.7%)
and 12 (24%) patients, and both simultaneously in three (10%) and three (6%) patients, respectively. Delayed graft function occurred only with deceased donors (nine patients, 30%). Patients with an
uneventful postoperative course in the deceased donor recipient group numbered nine (30%) and,
in the living donor recipient group, 27 (54%). Both α- and π-GST were significantly elevated at
1st postoperative day (POD) in deceased donor recipients, with acute rejection when compared
with the corresponding healthy subgroup (α-GST: Mean 473.5 ± 818 vs. 15.6 ± 21.2 ng/mg uCrea, 5 of 115 o J. Clin. Med. 2019, 8, 1899
n. Med. 2019, 8, x FOR PE p = 0.0094; π-GST: mean 477.8 ± 804 vs. 8 ± 6.4 ng/mg uCrea, p = 0.0023). In living donor recipients,
only π-GST showed an increase, reaching a peak at day 5 (mean 21.5 ± 28.2 ng/mg uCrea); however,
there was no significant difference between this group and the healthy subgroup. In patients with
CNI toxicity, α-GST performed better in both recipient groups; in deceased donor recipients, the mean
was 316 ± 704.5 ng/mg uCrea at 1 POD and was easily discriminated from the uneventful subgroup
(p = 0.06). Also, in the living donor group, a rise of α-GST to 68.9 ± 219 ng/mg uCrea was noted at day 5,
when CNI toxicity occurred. t week after transplantation.
3.2. α- and π-GST in Recipients When both acute rejection and CNI toxicity were recorded, neither α-
nor π-GST was able to distinguish those patients; however, it is worth mentioning that the number of
subjects in this subgroup was very low in our study. In the case of delayed graft function, present only
in deceased donor recipients in our study, α- as well as π-GST were elevated in the urine, with means
at 1st POD of 81.5 ± 201.3 and 151.6 ± 270.6 ng/mg uCrea, respectively. Only π-GST levels proved
significant when compared to the control subgroup (p = 0.036) (Figure 3). g
p
y
g
g
re was no significant difference between this group and the healthy subgroup. In patients w
I toxicity, α-GST performed better in both recipient groups; in deceased donor recipients,
an was 316 ± 704.5 ng/mg uCrea at 1 POD and was easily discriminated from the uneven
group (p = 0.06). Also, in the living donor group, a rise of α-GST to 68.9 ± 219 ng/mg uCrea w
ed at day 5, when CNI toxicity occurred. When both acute rejection and CNI toxicity w
orded, neither α- nor π-GST was able to distinguish those patients; however, it is wo
ntioning that the number of subjects in this subgroup was very low in our study. In the case
ayed graft function, present only in deceased donor recipients in our study, α- as well as π-G
re elevated in the urine, with means at 1st POD of 81.5 ± 201.3 and 151.6 ± 270.6 ng/mg uCr
pectively. Only π-GST levels proved significant when compared to the control subgroup (
36) (Figure 3). Figure 3. Urinary α- and π-GST levels in subgroups of deceased and living donor recipients during
he first week after transplantation; G1: Acute graft rejection (AGR), G2: Calcineurin-induced
nephrotoxicity (CNI), G3: Simultaneous acute graft rejection and calcineurin-induced nephrotoxicity
Figure 3. Urinary α- and π-GST levels in subgroups of deceased and living donor recipients during the
first week after transplantation; G1: Acute graft rejection (AGR), G2: Calcineurin-induced nephrotoxicity
(CNI), G3: Simultaneous acute graft rejection and calcineurin-induced nephrotoxicity (AGR + CNI),
G4: Delayed graft function (DGF), G5: Event-free (control). ure 3. Urinary α- and π-GST levels in subgroups of deceased and living donor recipients duri
first week after transplantation; G1: Acute graft rejection (AGR), G2: Calcineurin-induc
phrotoxicity (CNI), G3: Simultaneous acute graft rejection and calcineurin-induced nephrotoxic
Figure 3. t week after transplantation.
3.2. α- and π-GST in Recipients Urinary α- and π-GST levels in subgroups of deceased and living donor recipients during the
first week after transplantation; G1: Acute graft rejection (AGR), G2: Calcineurin-induced nephrotoxicity
(CNI), G3: Simultaneous acute graft rejection and calcineurin-induced nephrotoxicity (AGR + CNI),
G4: Delayed graft function (DGF), G5: Event-free (control). (AGR + CNI), G4: Delayed graft function (DGF), G5: Event-free (control). Furthermore, we performed ROC curve and survival analysis on the most outstand
markers at a specific point in the study. π-GST showed the most promising results in decea
nor recipients with acute graft rejection and delayed graft function at day 1 after transplantati
ients who developed a biopsy confirmed acute graft rejection within the first week a
nsplantation had significantly higher levels of urinary π-GST at POD 1. With an estimated cu
21.4 ng/mg uCrea, π-GST was able to distinguish the occurrence of AGR from a rejection-f
rse with 100% and 66.6% sensitivity and specificity, respectively (Figure 4). Similar reliability
Furthermore, we performed ROC curve and survival analysis on the most outstanding biomarkers
at a specific point in the study. π-GST showed the most promising results in deceased donor recipients
with acute graft rejection and delayed graft function at day 1 after transplantation. Patients who
developed a biopsy confirmed acute graft rejection within the first week after transplantation had
significantly higher levels of urinary π-GST at POD 1. With an estimated cutoffof 21.4 ng/mg uCrea,
π-GST was able to distinguish the occurrence of AGR from a rejection-free course with 100% and
66.6% sensitivity and specificity, respectively (Figure 4). Similar reliability for π-GST was observed in
the DGF subgroup; however, the estimated cutoffat POD 1 was slightly lower at 18.3 ng/mg uCrea
(sensitivity, 100%; specificity, 62.6%). Higher urinary π-GST levels could be seen in recipients with 6 of 11 J. Clin. Med. 2019, 8, 1899 delayed graft function; this was observed at POD1, POD 3 as well as POD 7. π-GST was not able to
differentiate between the causes of graft dysfunction in the early postoperative period (Figure 5). 2019, 8, x FOR PEER REVIEW
Clin. Med. 2019, 8, x FOR PEER REVIEW
6 of 1 4. Correlation between urinary π-GST levels in recipients of brain-dead donors’ gr
and probability of graft survival without acute graft rejection (AGR) during the firs
Figure 4. t week after transplantation.
3.2. α- and π-GST in Recipients Correlation between urinary π-GST levels in recipients of brain-dead donors’ grafts on
POD 1, and probability of graft survival without acute graft rejection (AGR) during the first week
after transplantation. Figure 4. Correlation between urinary π-GST levels in recipients of brain-dead donors’ grafts on
POD 1, and probability of graft survival without acute graft rejection (AGR) during the first week
after transplantation. Figure 4. Correlation between urinary π-GST levels in recipients of brain-dead donors’ grafts on levels in recipients of brain-dead donors’ grafts on Correlation between urinary π-GST levels in recipients of brain-dead donors’
nd probability of graft survival without acute graft rejection (AGR) during the f
Figure 4. Correlation between urinary π-GST levels in recipients of brain-dead donors’ grafts on
POD 1, and probability of graft survival without acute graft rejection (AGR) during the first week
after transplantation. OD 1, and probability of graft survival without acute graft rejection (AGR) during the first wee
ter transplantation. ,
p
y
g
g
j
(
)
g
ansplantation. Figure 5. Correlation between urinary π-GST levels in recipients of brain-dead donors’ grafts on
POD 1, 3, and 7; and probability of graft survival without delayed graft function (DGF) during the
Figure 5. Correlation between urinary π-GST levels in recipients of brain-dead donors’ grafts on POD
1, 3, and 7; and probability of graft survival without delayed graft function (DGF) during the first week
after transplantation. (
)
g ansplantation. on. igure 5. Correlation between urinary π-GST levels in recipients of brain-dead donors’ grafts o
OD 1, 3, and 7; and probability of graft survival without delayed graft function (DGF) during th
Figure 5. Correlation between urinary π-GST levels in recipients of brain-dead donors’ grafts on POD
1, 3, and 7; and probability of graft survival without delayed graft function (DGF) during the first week
after transplantation. first week after transplantat
3.3. α- and π-GST in Donors survival curve analysis (p 0.0109) (Figure 7). On the other hand, urinary GST in living donors showed no differences between subgro
the corresponding control group, and therefore failed to predict future events in recipients. Figure 6. Urinary α- and π-GST levels in deceased donors before transplantation; G1: Acute graf
rejection (AGR), G2: Calcineurin-induced nephrotoxicity (CNI), G3: Simultaneous acute graf
rejection and calcineurin-induced nephrotoxicity (AGR+CNI), G4: Delayed graft function (DGF), G5
Figure 6. Urinary α- and π-GST levels in deceased donors before transplantation; G1: Acute
graft rejection (AGR), G2: Calcineurin-induced nephrotoxicity (CNI), G3: Simultaneous acute graft
rejection and calcineurin-induced nephrotoxicity (AGR+CNI), G4: Delayed graft function (DGF), G5:
Event-free (control). ure 6. Urinary α- and π-GST levels in deceased donors before transplantation; G1: Acute
ction (AGR), G2: Calcineurin-induced nephrotoxicity (CNI), G3: Simultaneous acute
ction and calcineurin-induced nephrotoxicity (AGR+CNI), G4: Delayed graft function (DGF)
nt-free (control). d donors before transplantation; G1: Acute g
oto i ity (CNI)
G3
Si
ulta eou
a ute ure 6. Urinary α- and π-GST levels in deceas
ction (AGR)
G2: Calcineurin-induced neph ure 6. Urinary α- and π-GST levels in deceased donors before transplantation; G1: Acute gr
ction (AGR), G2: Calcineurin-induced nephrotoxicity (CNI), G3: Simultaneous acute gr
ction and calcineurin-induced nephrotoxicity (AGR+CNI), G4: Delayed graft function (DGF), G
Figure 6. Urinary α- and π-GST levels in deceased donors before transplantation; G1: Acute
graft rejection (AGR), G2: Calcineurin-induced nephrotoxicity (CNI), G3: Simultaneous acute graft
rejection and calcineurin-induced nephrotoxicity (AGR+CNI), G4: Delayed graft function (DGF), G5:
Event-free (control). n (AGR), G2: Calcineurin induced nephrotoxicity (CNI), G3: Simultaneous acute
n and calcineurin-induced nephrotoxicity (AGR+CNI), G4: Delayed graft function (DG
ree (control). Event-free (control). Figure 7. Correlation between urinary α-GST levels in brain-dead donors before transplantation an
probability of graft survival without acute graft rejection in corresponding recipients during the fir
re 7. Correlation between urinary α-GST levels in brain-dead donors before transplantation
ability of graft survival without acute graft rejection in corresponding recipients during the
Figure 7. Correlation between urinary α-GST levels in brain-dead donors before transplantation and
probability of graft survival without acute graft rejection in corresponding recipients during the first
week after transplantation. vent-free (control). ure 7. Correlation between urinary α-GST levels in brain-dead donors before transplantation
7. Correlation between urinary α-GST levels in brain-dead donors before transplantat
lity of graft survival without acute graft rejection in corresponding recipients during
Figure 7. first week after transplantat
3.3. α- and π-GST in Donors Correlation between urinary α-GST levels in brain-dead donors before transplantation and
probability of graft survival without acute graft rejection in corresponding recipients during the first
week after transplantation. probability of graft survival without a
week after transplantation.
ek after transplantation.
3.4. Six- and 12-Months Graft Survival p
Six- and 12-Months Graft Survival
We followed up recipients of the subgroups G1 (AGR) and G4 (DGF) at six and 12 mon
r transplantation, as correlations of α- and π-GST in those cohorts showed the most promis
l
b
l
f f
l
h
f d
and 12-Months Graft Survival
e followed up recipients of the subgroups G1 (AGR) and G4 (DGF) at six and 12 m
ansplantation, as correlations of α- and π-GST in those cohorts showed the most pro
However in subgroup G1 only one patient out of four lost the graft due to rec
We followed up recipients of the subgroups G1 (AGR) and G4 (DGF) at six and 12 months
after transplantation, as correlations of α- and π-GST in those cohorts showed the most promising
results. However, in subgroup G1 only one patient out of four lost the graft due to recruiting nephritis;
in subgroup G4 all three grafts were lost due to death not associated with graft function (cancer or
cardiac arrest). first week after transplantat
3.3. α- and π-GST in Donors re 5. Correlation between urinary π-GST levels in recipients of brain-dead donors’ graft
1 3 and 7; and probability of graft survival without delayed graft function (DGF) during
3. α- and π-GST in Donors
α- and π-GST showed interesting results when measured in deceased donor urine befor
rgan harvesting as they were elevated in those with poorer graft function after transplantation
nd seemed to predict a foreseeable event such as acute rejection or delayed graft function, which
as 3- to 8-fold more likely to occur than in those recipients with an uneventful course of treatment
α- and π-GST showed interesting results when measured in deceased donor urine before organ
harvesting as they were elevated in those with poorer graft function after transplantation, and seemed
to predict a foreseeable event such as acute rejection or delayed graft function, which was 3- to 8-fold
more likely to occur than in those recipients with an uneventful course of treatment. α-GST stood
out in the subgroup with both acute rejection and CNI toxicity (p = 0.02), while α- and π-GST were 7 of 11 J. Clin. Med. 2019, 8, 1899 remarkably higher (but not significantly so) in the DGF subgroup compared to the control group
(Figure 6). As for the predictive value of GST when measured in donor urine, α-GST stood out with
significant results in the AGR subgroup when ROC curve analysis was performed, and a cutoffvalue
of >33.97 ng/mg uCrea was calculated (AUC, 0.86; sensitivity, 77.7%; specificity, 100%). Based on the
donor’s urinary α-GST alone, all four renal grafts from deceased donors that showed acute rejection in
recipients were distinguished from those who had an AGR-free course in the survival curve analysis
(p = 0.0109) (Figure 7). in. Med. 2019, 8, x FOR PEER REVIEW
7 o
wed acute rejection in recipients were distinguished from those who had an AGR-free course
survival curve analysis (p = 0.0109) (Figure 7). d acute rejection in recipients were distinguished from those who had an AGR-free co
vival curve analysis (p = 0.0109) (Figure 7). n the other hand, urinary GST in living donors showed no differences between sub
corresponding control group, and therefore failed to predict future events in recipien (p
) (
g
)
On the other hand, urinary GST in living donors showed no differences between subgroups and
the corresponding control group, and therefore failed to predict future events in recipients. ,
hritis; in subg
is; in subgrou
4. Discussion ncer or cardiac arrest). Discussion
or cardiac arrest). ussion
The results of our prospective study, evaluating urinary α- and π-GST in deceased as well as
living kidney donors and their corresponding recipients as biomarkers for graft quality and function, 8 of 11 J. Clin. Med. 2019, 8, 1899 suggest the potential value of these enzymes. Previous studies showed the ability of urinary α- and
π-GST to predict acute renal damage in kidney graft recipients and demonstrated a release of these
enzymes in malfunctioning grafts [10–15]. However, none of these groups has compared the course of α-
and π-GST in donors as well as in corresponding recipients. In addition, we investigated the differences
in the markers in two settings, brain-dead/deceased and living organ donation. Our analyses reveal
that the determination of urinary π-GST concentration in deceased donor recipients, especially on the
first day after graft transplantation, could be valuable and indicative of kidney allograft function and
survival without AGR or DGF. Secondly, we found higher concentrations of α- and π-GST in the urine
of deceased kidney donors, whose grafts performed poorly in the corresponding recipients; α-GST
was able to predict AGR before transplantation. A determination between the causes of impaired
allograft function could not be reached by assessing urinary α- and π-GST alone, though it is unlikely
that in the complex setting of transplantation a single biomarker will reliably distinguish between the
pathogenesis of multifactorial elements; therefore, the proposed markers should be seen as an useful
tool in addition to established methods. Research studies investigating α- and π-GST in living donation are extremely limited: in our
review of the literature we only found one publication on this issue, and this concerned liver rather
than kidney transplantation [16]. Our findings showed lower concentrations of α- and π-GST in the
urine of living donor kidney recipients than in that of deceased donor kidney recipients. This was
observed in almost all subgroups, and especially on POD 1. Except for a notable, yet insignificant,
rise of π-GST in living donor recipients with AGR when compared to the control group, our results
find no further significances of urinary α- and π-GST in living donor recipients when harmful events
occurred. ,
hritis; in subg
is; in subgrou
4. Discussion Considering the superior organ quality and logistics in living donation transplantation,
as demonstrated by over half of living donor grafts surviving the first week after transplantation
event-free compared to 30% of deceased donor grafts, lower urinary concentrations of α- nor π-GST in
living donor transplantation are to be expected. Daemen et al. found a correlation between α-GST and
warm ischemic time in grafts from donation after cardiac death [17]. In our study neither α- nor π-GST
had a proven correlation with ischemic time, although it should be noted that a different type of graft
was investigated in the work mentioned. The toxic effect of CNI agents on renal grafts and its association with the excretion of α-GST has
been described in the past [10,18]. Our results failed to indicate such a correlation in deceased or living
donor recipients. This might be due to our definition of the toxic range of CNI serum levels. Therapeutic
and toxic serum levels of several drugs and especially immunosuppressants have been known to be
inconsistent and even overlapping [6,19]. In the 1990s, serum levels of tacrolimus in the early period
after transplantation were suggested to be below 20 ng/mL in order to avoid side effects, whereas later
on levels above 15 ng/mg were proven to be associated with a higher risk of toxicity [20,21]. On the
other hand, the risk of acute rejection is significantly higher when there are low concentrations of
the agent [22]. A helpful step would be to find the toxic serum level of immunosuppressants that is
agreed upon by transplant communities; currently, despite all efforts, this varies significantly between
transplant centers. Another concerning factor is the design of the study, which did not include daily
and therefore more precise surveillance on that matter. In order to investigate the correlation between
toxic exposure to immunosuppressants and the excretion of GST into the urine, a closer observation
with more frequent sample collection is required. DGF was observed only in deceased donor recipients in our study. It occurred with an incidence
of 30%, which is similar to the findings of a recent work by Willicombe et al. [23]. Risk factors
and characteristics of donors and recipients associated with DGF such as cold ischemic time and
donor age have been established in previous publications [4,5,24]. However, taking these factors into
consideration, it is to be expected that DGF is less common in living donation. ,
hritis; in subg
is; in subgrou
4. Discussion It has been demonstrated
that α-GST excretion would be increased in the case of DGF due to its location in the renal tubular
system and the association between DGF and proximal tubular necrosis [8,25]. On the other hand,
π-GST has been shown to be of predictive value in terms of the need for dialysis in a publication by 9 of 11 J. Clin. Med. 2019, 8, 1899 Seabra et al. including 245 patients with acute kidney injury [26]. Hall et al. investigated α- and
π-GST in a perfusate solution during machine perfusion of kidney allografts from deceased donors,
and suggested an independent association between π-GST and DGF [27]. Our findings demonstrated
the consistent significance of urinary π-GST in differentiating between DGF and normally functioning
grafts when measured in deceased donor recipients. This was observed at several time points of the
study and had a strong power of sensitivity and specificity. Little is known about the behavior of the
proteins under dialysis, so it is unclear whether α-GST is more dialyzable than π-GST or the other
way round. The sample collection from patients undergoing dialysis in our study did not occur with
respect to dialysis time, which should be seen as another possible disturbance factor. Further limitations of this study are the small number of patients in certain subgroups and the
overall high standard deviations. We distinguished well between the causes of impaired graft function
and took into consideration simultaneous events. The time frame of our observation was limited in
that it focused only on the first week after transplantation. We believe that multiple serial samples and
an extended study design would be beneficial in future projects. 5. Conclusions In summary, the elevation of urinary π-GST in deceased donor kidney recipients at day 1 after
kidney transplantation could be a helpful monitoring parameter, in addition to urinary output and
serum creatinine, to determine graft function in recipients. It might be indicative of acute rejection or
a need for dialysis. The measuring of urinary α- and π-GST should also be considered in deceased
donors as this seems to be of predictive value in terms of graft outcome and might help with assessing
allograft quality. Our findings reveal an association between urinary α-GST in deceased donors and
AGR in corresponding recipients. Thus, further investigation of α- and π-GST in a larger population
and daily sample collection should be considered. Although urinary α- and π-GST in living kidney
donation showed no relevant correlation with harmful events in our analyses, this is, to the best
of our knowledge, the first study demonstrating differences in biomarkers between deceased and
living kidney donation, so subsequent investigations will be needed in order to confirm or contradict
our findings. Author Contributions: Conceptualization, S.K. and V.S.; methodology, S.K., V.S. and B.G.; software, S.K., A.R.-S. and B.S.; validation, B.G., M.M. and B.S.; formal analysis, S.K. and A.R.-S.; investigation, S.K., B.G. and V.S.;
resources, A.P., V.S. and N.M.O.; data curation, S.K., B.G., M.M. and N.M.O.; writing—original draft preparation,
S.K.; writing—review and editing, J.G.B., M.H.M. and T.V.; visualization, S.K., M.H.M. and T.V.; supervision,
V.S., A.P. and J.G.B.; project administration, S.K. and V.S. All authors have read and approved the final version of
the manuscript. Acknowledgments: We acknowledge support from the Open Access Publication Fund of the University of
Münster. The authors also extend their gratitude to Argutus Medical Ltd. (Dublin, Ireland) for providing the kits. Conflicts of Interest: The authors declare no conflict of interest. Conflicts of Interest: The authors declare no conflict of interest. Author Contributions: Conceptualization, S.K. and V.S.; methodology, S.K., V.S. and B.G.; software, S.K., A.R.-S.
and B.S.; validation, B.G., M.M. and B.S.; formal analysis, S.K. and A.R.-S.; investigation, S.K., B.G. and V.S.;
resources, A.P., V.S. and N.M.O.; data curation, S.K., B.G., M.M. and N.M.O.; writing—original draft preparation,
S.K.; writing—review and editing, J.G.B., M.H.M. and T.V.; visualization, S.K., M.H.M. and T.V.; supervision,
V.S., A.P. and J.G.B.; project administration, S.K. and V.S. All authors have read and approved the final version of
the manuscript. References 1. Wu, W.K.; Famure, O.; Li, Y.; Kim, S.J. Delayed graft function and the risk of acute rejection in the modern
era of kidney transplantation. Kidney Int. 2015, 88, 851–858. [CrossRef] 1. Wu, W.K.; Famure, O.; Li, Y.; Kim, S.J. Delayed graft function and the risk of acute rejection in the modern
era of kidney transplantation. Kidney Int. 2015, 88, 851–858. [CrossRef] 2. Marcen, R.; Fernandez-Rodriguez, A.; Rodriguez-Mendiola, N.; Ponte, B.; Galeano, C.; Villafruela, J.J.;
Teruel, J.L.; Burgos, F.J.; Ortuno, J. Evolution of rejection rates and kidney graft survival: A historical analysis. Transplant Proc. 2009, 41, 2357–2359. [CrossRef] 2. Marcen, R.; Fernandez-Rodriguez, A.; Rodriguez-Mendiola, N.; Ponte, B.; Galeano, C.; Villafruela, J.J.;
Teruel, J.L.; Burgos, F.J.; Ortuno, J. Evolution of rejection rates and kidney graft survival: A historical analysis. Transplant Proc. 2009, 41, 2357–2359. [CrossRef] 3. Martins, L.; Ventura, A.; Branco, A.; Carvalho, M.J.; Henriques, A.C.; Dias, L.; Sarmento, A.M.; Amil, M. Cyclosporine versus tacrolimus in kidney transplantation: Are there differences in nephrotoxicity? Transplant Proc. 2004, 36, 877–879. [CrossRef] 3. Martins, L.; Ventura, A.; Branco, A.; Carvalho, M.J.; Henriques, A.C.; Dias, L.; Sarmento, A.M.; Amil, M. Cyclosporine versus tacrolimus in kidney transplantation: Are there differences in nephrotoxicity? Transplant Proc. 2004, 36, 877–879. [CrossRef] 4. Ojo, A.O.; Wolfe, R.A.; Held, P.J.; Port, F.K.; Schmouder, R.L. Delayed graft function: Risk factors and
implications for renal allograft survival. Transplantation 1997, 63, 968–974. [CrossRef] 4. Ojo, A.O.; Wolfe, R.A.; Held, P.J.; Port, F.K.; Schmouder, R.L. Delayed graft function: Risk factors and
implications for renal allograft survival. Transplantation 1997, 63, 968–974. [CrossRef] 5. Siedlecki, A.; Irish, W.; Brennan, D.C. Delayed graft function in the kidney transplant. Am. J. Transplant. 2011, 11, 2279–2296. [CrossRef] 5. Siedlecki, A.; Irish, W.; Brennan, D.C. Delayed graft function in the kidney transplant. Am. J. Transplant. 2011, 11, 2279–2296. [CrossRef] 10 of 11 J. Clin. Med. 2019, 8, 1899 6. Klintmalm, G.; Sawe, J.; Ringden, O.; von Bahr, C.; Magnusson, A. Cyclosporine plasma levels in renal
transplant patients. Association with renal toxicity and allograft rejection. Transplantation 1985, 39, 132–137. [CrossRef] 7. Wilczek, H.E. Percutaneous needle biopsy of the renal allograft. A clinical safety evaluation of 1129 b
Transplantation 1990, 50, 790–797. [CrossRef] 8. Branten, A.J.; Mulder, T.P.; Peters, W.H.; Assmann, K.J.; Wetzels, J.F. Urinary excretion of glutathione S
transferases alpha and pi in patients with proteinuria: Reflection of the site of tubular injury. Nephron 2000,
85, 120–126. [CrossRef] 9. References Harrison, D.J.; Kharbanda, R.; Cunningham, D.S.; McLellan, L.I.; Hayes, J.D. Distribution of glutathione
S-transferase isoenzymes in human kidney: Basis for possible markers of renal injury. J. Clin. Pathol. 1989,
42, 624–628. [CrossRef] 10. Backman, L.; Appelkvist, E.L.; Ringden, O.; Dallner, G. Glutathione transferase in the urine: A marker for
post-transplant tubular lesions. Kidney Int. 1988, 33, 571–577. [CrossRef] 11. Kuzniar, J.; Marchewka, Z.; Krasnowski, R.; Boratynska, M.; Dlugosz, A.; Klinger, M. Enzymuria and low
molecular weight protein excretion as the differentiating marker of complications in the early post kidney
transplantation period. Int. Urol. Nephrol. 2006, 38, 753–758. [CrossRef] 12. Polak, W.P.; Kosieradzki, M.; Kwiatkowski, A.; Danielewicz, R.; Lisik, W.; Michalak, G.; Paczek, L.; Lao, M.;
Walaszewski, J.; Rowinski, W.A. Activity of glutathione S-transferases in the urine of kidney transplant
recipients during the first week after transplantation. Ann. Transplant. 1999, 4, 42–45. 13. Sundberg, A.G.; Appelkvist, E.L.; Backman, L.; Dallner, G. Urinary Pi-Class Glutathione Transferase as an
Indicator of Tubular Damage in the Human Kidney. Nephron 1994, 67, 308–316. [CrossRef] 14. Backman, L.; Appelkvist, E.L.; Ringden, O.; Dallner, G. Appearance of basic glutathione transferase in the
urine during tubular complications in renal transplant recipients. Transplant. Proc. 1988, 20, 427–430. 15. Westhuyzen, J.; Endre, Z.H.; Reece, G.; Reith, D.M.; Saltissi, D.; Morgan, T.J. Measurement of tubular
enzymuria facilitates early detection of acute renal impairment in the intensive care unit. Nephrol. Dial. Transplant. 2003, 18, 543–551. [CrossRef] 16. Jochum, C.; Beste, M.; Sowa, J.P.; Farahani, M.S.; Penndorf, V.; Nadalin, S.; Saner, F.; Canbay, A.; Gerken, G. Glutathione-S-transferase subtypes alpha and pi as a tool to predict and monitor graft failure or regeneration
in a pilot study of living donor liver transplantation. Eur. J. Med. Res. 2011, 16, 34–40. [CrossRef] 17. Daemen, J.W.; Oomen, A.P.; Janssen, M.A.; van de Schoot, L.; van Kreel, B.K.; Heineman, E.; Kootstra, G. Glutathione S-transferase as predictor of functional outcome in transplantation of machine-preserved
non-heart-beating donor kidneys. Transplantation 1997, 63, 89–93. [CrossRef] 18. Sundberg, A.; Appelkvist, E.L.; Dallner, G.; Nilsson, R. Glutathione transferases in the urine—Sensitive
methods for detection of kidney damage-induced by nephrotoxic agents in humans. Environ. Health Perspect. 1994, 102, 293–296. 19. Min, D.I.; Perry, P.J.; Chen, H.Y.; Hunsicker, L.G. Cyclosporine trough concentrations in predicting allograft
rejection and renal toxicity up to 12 months after renal transplantation. Pharmacotherapy 1998, 18, 282–287. 20. Bottiger, Y.; Brattstrom, C.; Tyden, G.; Sawe, J.; Groth, C.G. © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/). References Tacrolimus whole blood concentrations correlate
closely to side-effects in renal transplant recipients. Br. J. Clin. Pharmacol. 1999, 48, 445–448. [CrossRef] 20. Bottiger, Y.; Brattstrom, C.; Tyden, G.; Sawe, J.; Groth, C.G. Tacrolimus whole blood concentrations correlate
closely to side-effects in renal transplant recipients. Br. J. Clin. Pharmacol. 1999, 48, 445–448. [CrossRef]
21. Schiff, J.; Cole, E.; Cantarovich, M. Therapeutic monitoring of calcineurin inhibitors for the nephrologist. closely to side-effects in renal transplant recipients. Br. J. Clin. Pharmacol. 1999, 48, 445–448. [CrossRef]
21. Schiff, J.; Cole, E.; Cantarovich, M. Therapeutic monitoring of calcineurin inhibitors for the nephrologist. Clin. J. Am. Soc. Nephrol. 2007, 2, 374–384. [CrossRef] 21. Schiff, J.; Cole, E.; Cantarovich, M. Therapeutic monitoring of calcineurin inhibitors for the nephrologist. Clin. J. Am. Soc. Nephrol. 2007, 2, 374–384. [CrossRef] 22. Undre, N.A.; van Hooff, J.; Christiaans, M.; Vanrenterghem, Y.; Donck, J.; Heeman, U.; Kohnle, M.; Zanker, B.;
Land, W.; Morales, J.M.; et al. Stevenson, P. Low systemic exposure to tacrolimus correlates with acute
rejection. Transplant. Proc. 1999, 31, 296–298. [CrossRef] 23. Willicombe, M.; Rizzello, A.; Goodall, D.; Papalois, V.; McLean, A.G.; Taube, D. Risk factors and outcomes of
delayed graft function in renal transplant recipients receiving a steroid sparing immunosuppression protocol. World J. Transplant. 2017, 7, 34–42. [CrossRef] [PubMed] 24. Irish, W.D.; Ilsley, J.N.; Schnitzler, M.A.; Feng, S.; Brennan, D.C. A risk prediction model for delayed graft
function in the current era of deceased donor renal transplantation. Am. J. Transplant. 2010, 10, 2279–2286. [CrossRef] 25. Sundberg, A.G.; Nilsson, R.; Appelkvist, E.L.; Dallner, G. Immunohistochemical Localization of Alpha-Class
and Pi-Class Glutathione Transferases in Normal Human Tissues. Pharmacol. Toxicol. 1993, 72, 321–331. [CrossRef] 11 of 11 J. Clin. Med. 2019, 8, 1899 26. Seabra, V.F.; Perianayagam, M.C.; Tighiouart, H.; Liangos, O.; dos Santos, O.F.; Jaber, B.L. Urinary alpha-GST
and pi-GST for prediction of dialysis requirement or in-hospital death in established acute kidney injury. Biomarkers 2011, 16, 709–717. [CrossRef] [PubMed] 27. Hall, I.E.; Bhangoo, R.S.; Reese, P.P.; Doshi, M.D.; Weng, F.L.; Hong, K.; Lin, H.; Han, G.; Hasz, R.D.;
Goldstein, M.J.; et al. Glutathione S-transferase iso-enzymes in perfusate from pumped kidneys are
associated with delayed graft function. Am. J. Transplant. 2014, 14, 886–896. [CrossRef] © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).
|
https://openalex.org/W4220880388
|
https://www.frontiersin.org/articles/10.3389/fcvm.2022.857019/pdf
|
English
| null |
Beat-Level Interpretation of Intra-Patient Paradigm Based on Object Detection
|
Frontiers in cardiovascular medicine
| 2,022
|
cc-by
| 8,725
|
ORIGINAL RESEARCH
published: 18 March 2022
doi: 10.3389/fcvm.2022.857019 Beat-Level Interpretation of
Intra-Patient Paradigm Based on
Object Detection
Man Kang 1,2, Xue-Feng Wang 1,2, Jing Xiao 3, He Tian 1,2* and Tian-Ling Ren 1,2*
1 The School of Integrated Circuits, Tsinghua University, Beijing, China, 2 The Beijing National Research Center for Information
Science and Technology, Tsinghua University, Beijing, China, 3 Ping An AI Research Center, Ping an Technology (Shenzhen)
Co. Ltd., Shenzhen, China 1 The School of Integrated Circuits, Tsinghua University, Beijing, China, 2 The Beijing National Research Center for Information
Science and Technology, Tsinghua University, Beijing, China, 3 Ping An AI Research Center, Ping an Technology (Shenzhen)
Co. Ltd., Shenzhen, China Electrocardiogram (ECG), as a product that can most directly reflect the electrical
activity of the heart, has become the most common clinical technique used for the
analysis of cardiac abnormalities. However, it is a heavy and tedious burden for doctors
to analyze a large amount of ECG data from the long-term monitoring system. The
realization of automatic ECG analysis is of great significance. This work proposes a
beat-level interpretation method based on the automatic annotation algorithm and object
detector, which abandons the previous mode of separate R peak detection and heartbeat
classification. The ground truth of the QRS complex is automatically annotated and also
regarded as the object the model can learn like category information. The object detector
unifies the localization and classification tasks, achieving an end-to-end optimization
as well as decoupling the high dependence on the R peak. Compared with most
advanced methods, this work shows superior performance. For the interpretation of 12
heartbeat types in the MIT-BIH dataset, the average accuracy is 99.60%, the average
sensitivity is 97.56%, and the average specificity is 99.78%. This method can be used
as a clinical auxiliary tool to help doctors diagnose arrhythmia after receiving large-scale
database training. Edited by:
Chunsheng Jiang,
China Academy of Engineering
Physics, China Physics, China
Reviewed by:
Ziyu Liu,
Fudan University, China
Huajing Fang,
Xi’an Jiaotong University, China
*Correspondence:
Tian-Ling Ren
tianhe88@tsinghua.edu.cn
He Tian
rentl@tsinghua.edu.cn Reviewed by:
Ziyu Liu,
Fudan University, China
Huajing Fang,
Xi’an Jiaotong University, China *Correspondence:
Tian-Ling Ren
tianhe88@tsinghua.edu.cn
He Tian
rentl@tsinghua.edu.cn tianhe88@tsinghua.edu.cn
He Tian Keywords: object detection, ECG, beat-level classification, deep learning, automatic annotation Specialty section:
This article was submitted to
Cardiac Rhythmology,
a section of the journal
Frontiers in Cardiovascular Medicine
Received: 18 January 2022
Accepted: 04 February 2022
Published: 18 March 2022 Specialty section:
This article was submitted to
Cardiac Rhythmology,
a section of the journal
Frontiers in Cardiovascular Medicine Specialty section:
This article was submitted to
Cardiac Rhythmology,
a section of the journal
Frontiers in Cardiovascular Medicine INTRODUCTION The WHO report regards cardiovascular disease as one of the leading causes of death worldwide,
and it will continue to exist in the near future. The burden of cardiovascular disease is so heavy
that research on heart health has to be taken seriously. Electrocardiogram (ECG), as a product that
can most directly reflect the electrical activity of the heart, has become the most common clinical
technique used for the analysis of cardiac abnormalities. However, it is a heavy and tedious burden
for doctors to analyze a large amount of ECG data from the long-term monitoring system. It is
necessary to realize automatic ECG analysis. Received: 18 January 2022
Accepted: 04 February 2022
Published: 18 March 2022 Keywords: object detection, ECG, beat-level classification, deep learning, automatic annotation Heartbeat Classification Heartbeat Classification
According to different feature extraction methods, heartbeat
classification can be divided into artificial methods and automatic
methods. Common artificial features include morphological
features (6–8) such as RR interval, and ECG transform
coefficients (9–13) such as wavelet transform. These features are
sent to a traditional classifier for interpretation. For example,
Hu (14) extracted features based on multiple discriminant and
principal component analysis, and used support vector machine
(SVM) for classification. Song et al. used linear discriminant
analysis combined with SVM to analyze six types of arrhythmias
(15). Melgani and Bazi proposed an SVM classifier based on
particle swarm optimization (16). Martis et al. used a four-layer
feedforward neural network and a least squares support vector
machine (LS-SVM) to divide heart beats into five categories
(17). Ganeshkumar and Kumaraswamy introduced a random
forest tree (RFT) as a classifier (18). Park et al. proposed a
K-nearest neighbor (K-NN) classifier (19). Jun et al. proposed
a parallel K-NN classifier for high-speed arrhythmia detection
(20). The accuracy of the above methods highly depends on
the effectiveness of feature extraction, which requires strong
scientific theories and doctors’ personal experience as support,
as well as the computational complexity is high. With the
development and application of deep learning technology, it has
also become a research hotspot in the field of ECG classification. In some previous studies, the simple 1D-CNN was used to
classify a time series of ECG signals (21–23) or one-dimensional
heartbeats (24). Acharya et al. proposed a nine-layer deep CNN
(25), which can identify five different heartbeats. Chauhan and
Vig used the deep long short-term memory (LSTMs) network
to classify abnormal and normal signals (26). Eltrass et al. (27)
proposed the CQ-NSGT algorithm, a new method for converting
one-dimensional signals into time-frequency maps, and used
AlexNet for time-frequency maps classification. Warrick and
Homosi proposed a new method to automatically classify
arrhythmias in ECG, using a combination of CNN and LSTM
(28). Shu et al. also proposed a system with combination of
CNN and LSTM (29) to identify five heart beats. Wong et al. (30) proposed a FPGA implementation of ECG classifier based
on bCNN, the core of which is to reduce the computational
complexity of CNN. Yao et al. (31) proposed an integrated CNN PROPOSED METHOD Figure 1B shows the overall framework of the ECG analysis
method in this work. There are two key designs: Automatic
heartbeats annotation and Object detector. Citation: Kang M, Wang X-F, Xiao J, Tian H and
Ren T-L (2022) Beat-Level
Interpretation of Intra-Patient
Paradigm Based on Object Detection. Front. Cardiovasc. Med. 9:857019. doi: 10.3389/fcvm.2022.857019 Kang M, Wang X-F, Xiao J, Tian H and
Ren T-L (2022) Beat-Level
Interpretation of Intra-Patient
Paradigm Based on Object Detection. Front. Cardiovasc. Med. 9:857019. doi: 10.3389/fcvm.2022.857019 In the past few decades, the open source ECG databases have promoted the development of
automatic ECG analysis. Most methods are based on one-dimensional ECG signals, as shown
in Figure 1A, which mainly involve four steps: signal preprocessing, R-peak detection, feature
extraction, and classifier construction. March 2022 | Volume 9 | Article 857019 Frontiers in Cardiovascular Medicine | www.frontiersin.org Beat-Level Interpretation of Intra-Patient Paradigm Based on Object Detection Kang et al. Signal Preprocessing The ECG signal has some interference such as baseline drift and
high-frequency noise. In order to obtain a better detection effect
in the follow-up, it is necessary to preprocess the signal in the
early stage. R-Peak Detection and GRU classifier to classify a time series of ECG signals. Sarin
et al. (32) compared the classification accuracy of MLP, CNN, and
LSTM on a subset of the MIT-BIH dataset. Over the years, a large number of studies have made efforts
in QRS complex detection. Pan and Tompkins (1) realized the
automatic detection of the R-peak through the first derivative,
non-linear transformation and amplitude/noise threshold. Using
time-domain features, Yeh and Wang (2) proposed a differential
operation method (DOM) algorithm. Li et al. (3) introduced
wavelet transform to the automatic detection of R-peak for the
first time. Martinez et al. (4) remove the singularity analysis,
and consider any possible QRS complex wave shape, search for
positive and negative zero crossing points. Manikandan and
Soman (5) introduced a new method using Shannon energy
estimation and Hilbert transform. These methods are time-
consuming, which require complex mathematical calculations, as
well as the accuracy is not always ideal. Most previous works just classify a signal over a long
period of time to get simple information about the existence of
abnormality. There are many types of heartbeats in a section of
ECG signal such as bigeminy and trigeminy, so this work focuses
on the interpretation of beat-level to get detailed information
about each beat for further analysis. Beat-level classification
in the past required the R-peak detection before classification
especially the inference stage also cost the same as the training
mode. More importantly, the classification accuracy was highly
dependent on R-peak detection quality and the independence
of R-peak detection made end-to-end optimization impossible. In this work, the position of QRS complex is also taken as the
object to be learned by the model-like category information,
which unifies the positioning and classification tasks based on
the object detector, achieving an end-to-end overall optimization
without independent time-consuming R-peak detection process. Besides, we expect the model to classify by learning features
from the morphology of heartbeats, much like doctor’s eyes,
as shown in Figure 1B, this work analyzes the original two-
dimensional images of heartbeats, not performing a series of
complex mathematical calculations for one-dimensional signals
or time-frequency diagram. Power Frequency Interference According to the standards published by the American College
of Cardiology (ACC), for the normal ECG signal, the amplitude
range is between 0.05 and 5 mV, the frequency range is
within the range of 0.05–100 Hz, and the spectral energy is
generally concentrated between 0.25 and 35 Hz. This work uses a
Butterworth filter with a cutofffrequency of 45 Hz and order of 10
to remove power frequency interference. In order to evaluate the
effect of noise reduction, we add a 50 Hz noise signal to simulate
power frequency interference. Figure 2 shows the comparison of
the effect before and after the noise signal processing. Frontiers in Cardiovascular Medicine | www.frontiersin.org Baseline Drift Baseline
drift
is
generally
caused
by
human
breathing
and electrode movement, which belongs to low-frequency
interference, usually below 0.5 Hz. The ST-segment frequency
band is in the range of 0.7–2.0 Hz, which partially overlaps with
the baseline drift frequency band, 0.05–1.5 Hz. It is necessary
to avoid obvious deformation of low frequency parts such Frontiers in Cardiovascular Medicine | www.frontiersin.org Frontiers in Cardiovascular Medicine | www.frontiersin.org March 2022 | Volume 9 | Article 857019 2 Beat-Level Interpretation of Intra-Patient Paradigm Based on Object Detection Kang et al. FIGURE 1 | The overview of the framework for analysis of ECG abnormality. (A) The framework of traditional method for ECG analysis. (B) The framework of
proposed method for ECG analysis. FIGURE 1 | The overview of the framework for analysis of ECG abnormality. (A) The framework of traditional method for ECG analysis. (B) The framework of
proposed method for ECG analysis. the framework for analysis of ECG abnormality. (A) The framework of traditional method for ECG analysis. (B) The framework of
nalysis FIGURE 1 | The overview of the framework for analysis of ECG abnormality. (A) The framework of traditional method for ECG analysis. (B) The framework of
proposed method for ECG analysis. FIGURE 2 | Power frequency interference reduction. (Left) Time domain. (Right) Frequency domain. FIGURE 2 | Power frequency interference reduction. (Left) Time domain. (Right) Frequency domain. Frontiers in Cardiovascular Medicine | www.frontiersin.org March 2022 | Volume 9 | Article 857019 3 Beat-Level Interpretation of Intra-Patient Paradigm Based on Object Detection Kang et al. FIGURE 3 | Baseline drift removing. (Left) Time domain. (Right) Frequency domain. FIGURE 3 | Baseline drift removing. (Left) Time domain. (Right) Frequency domain. FIGURE 3 | Baseline drift removing. (Left) Time domain. (Right) Frequency domain. FIGURE 3 | Baseline drift removing. (Left) Time domain. (Right) Frequency domain. FIGURE 4 | Automatic heartbeats annotation. FIGURE 4 | Automatic heartbeats annotation. FIGURE 4 | Automatic heartbeats annotation. FIGURE 4 | Automatic heartbeats annotation. Frontiers in Cardiovascular Medicine | www.frontiersin.org March 2022 | Volume 9 | Article 857019 4 Beat-Level Interpretation of Intra-Patient Paradigm Based on Object Detection Kang et al. FIGURE 5 | The framework of object detector. FIGURE 5 | The framework of object detector. We select the wavelet coefficients at j = 1 scale (34), and the
officially marked R peaks are used as the reference to locate Q-
wave and S-wave. Baseline Drift The R peak corresponds to the zero-crossing
point of the modulus maximum–minimum pair, Q-wave and S-
wave are located at the modulus minimum and maximum within
a certain range before and after the R peak, respectively. We need
to give certain delay compensation during the actual operation
because each waveform is not completely symmetric. Figure 4
shows the detailed process. as ST segment, or which will lead to detection and analysis
distortion. Therefore, the median filter, which has a good effect
on suppressing larger drift as well as protecting smaller P-wave
and T-wave, is adopted. The effect of drift removal mainly depends on the filter
window width. If the width is too small, the fitted baseline
has more high-frequency components, while too large width
causes too much computation and affects speed of the algorithm. Considering both speed and effect, as well as for convenience of
value, the width is set to half of the sampling rate. We use a rectangular box to annotate the position of QRS
complex, with Q-wave position as the left boundary, S-wave
position as the right boundary, R-peak position as the upper
boundary, and the smaller ordinate in Q-wave and S-wave as the
lower boundary. The upper left corner (xmin, ymin) and lower
right corner (xmax, ymax) are saved as location information in
.xml file format. Besides, in order to avoid edge effects, the signal is expanded
before filtering. Specifically, the endpoint value instead of 0 is
added to both ends of the signal to avoid larger errors. Figure 3 shows the effects before and after the processing of
a signal with severe baseline drift in the MIT-BIH database (see
section MIT-BIH Database). It can be seen that the drift is well-
suppressed, and only the very low-frequency part is attenuated
while the high-frequency part is barely affected. Automatic Heartbeats Annotation After the signal preprocessing described in section Signal
Preprocessing, the heartbeat images are saved in.jpg format. In
order to take the position information as the object that can be
learned by the subsequent model, it is necessary to annotate the
location of the heartbeats. Frontiers in Cardiovascular Medicine | www.frontiersin.org Object Detector We use Cascade RCNN (35). as a basic model, and Figure 5 shows
the overall framework of the model. Region Proposal Network Region proposal network takes an image as input and outputs
in a set of rectangular boxes called proposals. Each proposal has
a score, which measures the confidence belongs to foreground. As shown in Figure 6, in order to generate a region proposal,
we slide a small window of n∗n (n = 3 in this work) on the
convolutional feature map. Each sliding window is mapped to
a lower-dimensional feature that is fed into two fully connected
layer branches: a regression branch (reg) and a classification
branch (cls). Since it operates in a sliding window mode, the fully
connected layers are shared in all spatial locations. The transient degree of signal is often described by singularity. Wavelet transform is an effective method to analyze signal
singularity, which has promising features for characterizing
the local regularity of signals by decomposing the signal into
elementary building blocks that are well-localized both in
time and frequency (33). Each pair of positive and negative
modulus maximum of the wavelet coefficient corresponds to
a signal singularity, and the zero-crossing point between the
pair is the singularity position. The relationship between wavelet
decomposition and singularity varies in different scales. The
small scale reflects the high-frequency component of the signal,
while the large scale corresponds to the opposite. At each sliding window position, we predict multiple region
proposals at the same time, where the maximum number of
proposals at each position is denoted as k (k = 3). Therefore,
the reg branch has 4k outputs to encode the coordinates of
the k rectangular boxes, and the cls branch outputs 2k scores
to estimate the foreground or background probability of each
proposal. The k proposals are parameterized with respect to k
reference boxes called anchors. The anchors center on the sliding
window and are associated with scale and aspect ratio. In order Different bands could be located by wavelet decomposition at
different scales due to an ECG signal varies in frequency. Q-wave
and S-wave are typical high-frequency waves with low amplitude,
whose energy is concentrated in small scale. March 2022 | Volume 9 | Article 857019 5 Beat-Level Interpretation of Intra-Patient Paradigm Based on Object Detection Kang et al. FIGURE 6 | Region proposal network (RPN). FIGURE 6 | Region proposal network (RPN). to reduce redundancy, Non-maximum suppression (NMS) is
used for proposals according to their scores. Then the proposals
with top scores are selected for follow-up detection. Region Proposal Network The NMS is
explained in detail later. previous stage, and the convolution feature map of a certain layer
or several layers. As shown in Figure 7, the RoI pooling layer maps the
coordinates to the corresponding position in the feature map. Each RoI is a rectangular area in the convolutional feature
map, which is defined by a quadruple (x, y, h, w), respectively,
corresponding to its center point coordinates (x, y), height h,
and width w. RoI pooling divides the RoI of h∗w into H∗W
grids consisted of sub-windows of approximate size h/H∗w/W,
and performs maximum pooling operation on each sub-window. Pooling is applied to each feature map channel independently, Region of Interest Pooling Region of interest pooling uses maximum pooling to transform
the features in RoI into a small feature map of fixed size H∗W
(7∗7), where H and W are hyperparameters independent of any
specific RoI. The inputs of RoI pooling are coordinate information
of the proposals from RPN or that of the predicted boxes from March 2022 | Volume 9 | Article 857019 Frontiers in Cardiovascular Medicine | www.frontiersin.org 6 Beat-Level Interpretation of Intra-Patient Paradigm Based on Object Detection Kang et al. FIGURE 7 | Region of interest (RoI) pooling. TABLE 1 | Categories and numbers of beats in the MIT-BIH database. AAMI classes
MIT-BIH
Annotations
Description
Numbers
Normal(N)
N
Normal beat
75,052
L
Left bundle branch
block beat
8,075
R
Right bundle branch
block beat
7,259
Supraventricular
ectopic beat
(SVEB)
E
Atrial escape beat
16
J
Nodal (junctional)
escape beat
229
A
Atrial premature beat
2,546
A
Aberrated atrial
premature beat
150
J
Nodal (junctional)
premature beat
83
S
Supraventricular
premature or ectopic
beat (atrial or nodal)
2
Ventricular ectopic
beat (VEB)
V
Premature ventricular
contraction
7,130
E
Ventricular escape beat
106
Fusion (F)
F
A fusion of ventricular
and normal beat
803
Unknown beat (Q)
/
Paced beat
7,028
F
A fusion of paced and
normal beat
982
Q
Unclassifiable beat
33
just like standard maximum pooling, thus we get fixed size
feature maps from RoIs of varying sizes. TABLE 2 | Categories and numbers of beats. Classes
Training set
Testing set
N
4,013
987
L
3,992
1,008
R
3,975
1,025
e
3,994
1,006
j
3,971
1,029
A
4,053
947
a
4,031
969
J
3,985
1,015
S
3,957
1,043
V
4,002
998
E
4,014
986
F
4,013
987
Total
48,000
12,000
TABLE 3 | The results of each category on testing set. Classes
Acc (%)
Sen (%)
Spe (%)
N
99.19
93.52
99.70
L
99.94
99.60
99.97
R
99.47
96.20
99.77
e
99.92
99.90
99.92
j
99.47
96.99
99.70
A
98.87
93.14
99.36
a
99.87
99.07
99.94
J
99.68
98.23
99.81
S
99.81
99.81
99.81
V
99.49
96.59
99.75
E
99.98
99.90
99.98
F
99.5
97.77
99.66 FIGURE 7 | Region of interest (RoI) pooling. FIGURE 7 | Region of interest (RoI) pooling. FIGURE 7 | Region of interest (RoI) pooling. TABLE 1 | Categories and numbers of beats in the MIT-BIH database. Region of Interest Pooling AAMI classes
MIT-BIH
Annotations
Description
Numbers
Normal(N)
N
Normal beat
75,052
L
Left bundle branch
block beat
8,075
R
Right bundle branch
block beat
7,259
Supraventricular
ectopic beat
(SVEB)
E
Atrial escape beat
16
J
Nodal (junctional)
escape beat
229
A
Atrial premature beat
2,546
A
Aberrated atrial
premature beat
150
J
Nodal (junctional)
premature beat
83
S
Supraventricular
premature or ectopic
beat (atrial or nodal)
2
Ventricular ectopic
beat (VEB)
V
Premature ventricular
contraction
7,130
E
Ventricular escape beat
106
Fusion (F)
F
A fusion of ventricular
and normal beat
803
Unknown beat (Q)
/
Paced beat
7,028
F
A fusion of paced and
normal beat
982
Q
Unclassifiable beat
33
just like standard maximum pooling, thus we get fixed size
feature maps from RoIs of varying sizes. Head
The Head layer is responsible for further processing the fixed
size feature maps from RoI pooling to output the final detection
results. Each feature map will go through a series of fully
connected layers, and finally branch into two output layers,
classification and regression. For each RoI, the classification layer
TABLE 2 | Categories and numbers of beats. Classes
Training set
Testing set
N
4,013
987
L
3,992
1,008
R
3,975
1,025
e
3,994
1,006
j
3,971
1,029
A
4,053
947
a
4,031
969
J
3,985
1,015
S
3,957
1,043
V
4,002
998
E
4,014
986
F
4,013
987
Total
48,000
12,000
TABLE 3 | The results of each category on testing set. Classes
Acc (%)
Sen (%)
Spe (%)
N
99.19
93.52
99.70
L
99.94
99.60
99.97
R
99.47
96.20
99.77
e
99.92
99.90
99.92
j
99.47
96.99
99.70
A
98.87
93.14
99.36
a
99.87
99.07
99.94
J
99.68
98.23
99.81
S
99.81
99.81
99.81
V
99.49
96.59
99.75
E
99.98
99.90
99.98
F
99.5
97.77
99.66
Average
99.60
97.56
99.78
outputs the softmax probabilities of C foreground classes and
one background class, while the regression layer outputs four real
numbers encoding predicted boxes position. Frontiers in Cardiovascular Medicine | www.frontiersin.org
7
March 2022 | Volume 9 | Article 857019 TABLE 1 | Categories and numbers of beats in the MIT-BIH database. TABLE 3 | The results of each category on testing set. outputs the softmax probabilities of C foreground classes and
one background class, while the regression layer outputs four real
numbers encoding predicted boxes position. Non-Maximum Suppression Non-maximum suppression, which is usually used in the RPN
and inference stage, aims to extract predicted boxes with
high confidence and suppresses the predicted boxes with low
confidence, thus could remove redundant boxes detecting the
same object. g
A bounding box is encoded as (45), g
A bounding box is encoded as (45), Before describing NMS, it is necessary to explain the IoU,
which measures the overlap between two boxes. As shown in
Figure 8, the IoU of box A and box B is tx = (x −xa)/wa,
ty = (y −ya)/ha,
tw = log(w/wa),
th = log(h/ha),
t∗
x = (x∗−xa)/wa,
t∗
y = (y∗−ya)/ha,
t∗
w = log(w∗/wa),
t∗
h = log(h∗/ha)
(1) Figure 8, the IoU of box A and box B is (1) IoU = A T B
A S B
(7) (7) where x, y, w, h are the center point’s coordinates, width, and
height of a rectangular box, x, xa, x∗, respectively, correspond to
the predicted box, anchor and ground truth, y, w, h are similar. Obviously, the higher the IoU value, the higher the degree of
overlap between two boxes. The location information we annotate are upper left corner
(xmin, ymin) and lower right corner (xmax, ymax) of a
rectangular box. So before calculating the regression loss, we
convert the coordinates to a quadruple (x, y, h w), where For an image, the final classification branch and regression
branch will output confidence scores and coordinate information
of a series of boxes. The specific process of the NMS is as follows. B represents the box set. First, selecting the predicted box M
with the maximum confidence score and adding it to the set D
representing final results. Second, removing the box M and other
category boxes whose IoU with M exceed preset threshold from
B. Next, repeating the two steps until B is empty. In this work,
only the boxes with the highest confidence score can be regarded
as the final predicted results in NMS because there is no overlap
between different heartbeat objects. x=(xmin+xmax)/2,
y=(ymin+ymax)/2,
w=xmax-
xmin, h=ymax-ymin, x=(xmin+xmax)/2,
y=(ymin+ymax)/2,
w=xmax-
xmin, h=ymax-ymin, Then the four values are encoded by Equation (1), which are
used as ground truth labels. The regression loss of a predicted box is defined as Lreg(t, t∗) =
X
i∈{x,y,w,h}
smoothL1(ti −t∗
i )
(2) (2) Loss Function In order to calculate the loss of each predicted box, we need to
classify them as foreground or background according to their
Intersection over Union (IoU) with ground truths. IoU will be
described in detail later. The total loss function is the sum of the three stages L = L1 + L2 + L3
(6) (6) Given an IoU threshold IoU_thres, the predicted boxes which
have maximum IoU with ground truths and the predicted boxes
whose IoU with any ground truth greater than IoU_thres are
regarded as foregrounds, while the predicted boxes whose IoU
with all ground truths less than IoU_thres are backgrounds. For
Stage1, Stage2, and Stage3 in Figure 5, the value of IoU_thres
is increasing. Non-Maximum Suppression Head The Head layer is responsible for further processing the fixed
size feature maps from RoI pooling to output the final detection
results. Each feature map will go through a series of fully
connected layers, and finally branch into two output layers,
classification and regression. For each RoI, the classification layer outputs the softmax probabilities of C foreground classes and
one background class, while the regression layer outputs four real
numbers encoding predicted boxes position. March 2022 | Volume 9 | Article 857019 Frontiers in Cardiovascular Medicine | www.frontiersin.org 7 Beat-Level Interpretation of Intra-Patient Paradigm Based on Object Detection Kang et al. TABLE 4 | The performance of our proposed method compared with previous
work. Work
Classes_n
Acc (%)
Sen (%)
Spe (%)
Zhou et al. (36)
4
98.51
94.41
98.45
Hou et al. (37)
5
99.45
98.63
99.66
Wan et al. (38)
5
99.1
–
–
Ullah et al. (39)
8
99.11
97.91
99.61
Wang (40)
2
97.4
97.9
97.1
Chen et al. (41)
6
99.32
97.75
99.51
Niu et al. (42)
3
96.4
–
–
Houssein et al. (43)
4
98.26
97.43
–
Naz et al. (44)
4
97.6
–
–
This work
12
99.60
97.56
99.78 TABLE 4 | The performance of our proposed method compared with previous
work. The classification loss is calculated by cross entropy, which is
defined as Lcls(p, c) = −log pc
(4) (4) where p is the softmax probability vector from the classification
branch, c is the true category label, taking 0, 1, 2...C. C is
the number of foreground classes, and 0 corresponds to the
background class. The loss function of each stage is defined as (35). Ls =
X
n
Lcls(ps,n, p∗
s,n) + λ
X
n
p∗
s,nLreg(ts,n, t∗
s,n)
(5) (5) where. balances the weight of classification loss and regression
loss, only the boxes belong to foreground class p∗
s,n = 1 need to
be calculated for regression losses. EXPERIMENTS AND RESULTS where where MIT-BIH Database MIT-BIH arrhythmia database, one of the four internationally
recognized standard ECG databases, is used for experimental
evaluation in this work. The database contains a total of 48 smoothL1(x) =
0.5x2, if |x| < 1
|x| −0.5, otherwise (3) Frontiers in Cardiovascular Medicine | www.frontiersin.org March 2022 | Volume 9 | Article 857019 Beat-Level Interpretation of Intra-Patient Paradigm Based on Object Detection Kang et al. TABLE 5 | Comparison of detection performance between Faster RCNN and Cascade RCNN. Faster RCNN
Cascade RCNN
Class
gts
dets
recall
AP
dets
recall
AP
N
943
948
91.7
90.5
930
92.5
91.4
SVEB
934
931
92.9
91.6
937
95.0
94.1
VEB
882
950
97.7
97.0
895
96.9
96.3
F
238
217
83.6
83.1
235
88.7
87.8
mAP@0.5
90.5
92.4
gts, the number of ground truths; dets, the number of objects the model detects; recall, the ratio of true positive objects detected to all positive objects; AP, average precision of single
category, calculated by PR curve; mAP@0.5, mean Average Precision (IoU = 0.5). TABLE 5 | Comparison of detection performance between Faster RCNN and Cascade RCNN. gts, the number of ground truths; dets, the number of objects the model detects; recall, the ratio of true positive objects detected to all positive objects; AP, average precision of single
category, calculated by PR curve; mAP@0.5, mean Average Precision (IoU = 0.5). FIGURE 8 | Intersection over union (IoU). samples, and perform data augmentation by translating and
resizing for various heartbeat images. A total of 60,000 heartbeat
images are used in the final experiment, which are divided into
training sets and testing sets in a ratio of 8:2. Table 2 shows the
numbers for each category in detail. Evaluation Metrics Consistent with previous research, classification accuracy(Acc),
sensitivity(Sen), and specificity(Spe) are used as evaluation
metrics in this work. Acc =
TP + TN
TP + FN + FP + TN
(8) (8) Sen =
TP
TP + FN
(9)
Spe =
TN
TN + FP
(10) Sen =
TP
TP + FN
(9) Sen =
TP
TP + FN
(9) (9) FIGURE 8 | Intersection over union (IoU). (10) records from 47 different patients, and the ECG signals are
sampled at 360 Hz with 11-bit resolution. The above metrics are applicable to each heartbeat category,
where TP is the number of heartbeats correctly classified as
positive samples, TN is the number of heartbeats correctly
classified as negative samples, FP is the number of heartbeats
incorrectly classified as positive samples, and FN is the number
of heartbeats incorrectly classified as negative samples. Each record corresponds to about 30 min of ECG data,
including signals from two leads. For all records, the first lead
is the modified lead II (MLII) and the second lead is V1, V2, V4,
or V5. Twenty-three records are randomly selected from 4,000
continuous 24-h dynamic ECG signals of patients at Beth Israel
Hospital (BIH), while the remaining data are selected from a few
rare ECG data samples and have important clinical significance. The notes are authoritatively certified by multiple cardiologists. Frontiers in Cardiovascular Medicine | www.frontiersin.org Implementation Details We use the Cascade RCNN equipped with Feature Pyramid
Network (FPN) (47) as our default framework, the ImageNet pre-
trained ResNet-50 (48) is adopted as the backbone, and the RoI
Align (49) is used to replace the RoI pooling. Our implementation
and hyperparameters are based on MMDetection (50). Anchors
with 1 scale and 3 aspect ratios are used. NMS with a threshold
of 0.7 is used to generate 2,000 and 1,000 proposals for training
and inference. In each training step, 512 proposals are sampled
from 2,000 proposals for training, and the ratio of foreground to
background proposals is 1:3. The
Association
for
the
Advancement
of
Medical
Instrumentation (AAMI) (46) divides the heartbeats in the
MIT-BIH database into five categories. Table 1 shows the specific
categories and numbers. Given MLII is the only lead representing all records in this
database, as well as the most commonly used lead for experts to
analyze, this work only extracts the ECG signals of MLII lead for
experiments. According to AAMI’s recommendation, the rhythm
heartbeat records, i.e., 102, 104, 107, and 217, are not used, and
the Q category which does not actually exist is also ignored. In order to balance the number of different heartbeat
categories, we only sample a part of Normal heartbeats as The model is trained for 24 epochs on 4 GPUs with 4 images
per GPU. With SGD optimizer, the learning rate is initialized to Frontiers in Cardiovascular Medicine | www.frontiersin.org March 2022 | Volume 9 | Article 857019 9 Beat-Level Interpretation of Intra-Patient Paradigm Based on Object Detection Kang et al. FIGURE 9 | Detection results of the proposed model on various heartbeat images. (A) RBBB beat. (B) Normal beat. (C) LBBB beat. (D) Nodal escape beat. (E) Nodal
premature beat. (F) Fusion of ventricular and normal beat. (G) Atrial escape beat. (H) Ventricular escape beat. (I) Atrial premature beat. (J) Aberrated atrial premature
beat. (K) Premature ventricular contraction. (L) Premature or ectopic supraventricular beat. FIGURE 9 | Detection results of the proposed model on various heartbeat images. (A) RBBB beat. (B) Normal beat. (C) LBBB beat. (D) Nodal escape beat. (E) Nodal
premature beat. (F) Fusion of ventricular and normal beat. (G) Atrial escape beat. (H) Ventricular escape beat. (I) Atrial premature beat. (J) Aberrated atrial premature
beat. (K) Premature ventricular contraction. (L) Premature or ectopic supraventricular beat. DISCUSSION Traditional ECG analysis methods require independent QRS
complex detection that causes that the effectiveness of feature
extraction and classification are highly dependent on the earlier
detection accuracy. Error detection and missing detection can
have a negative impact on feature extraction and eventually cause
wrong classification results. The Importance of Object Detector The human visual system is fast and accurate. At a glance, you
can immediately know what the objects in the image are, where
they are, and how they interact. We apply computer vision
knowledge to ECG abnormality analysis, and use the object
detection framework to directly predict the bounding boxes and
class probabilities from the heartbeat images. During training
and testing, the model can see the entire heartbeat image, so it Implementation Details FIGURE 9 | Detection results of the proposed model on various heartbeat images. (A) RBBB beat. (B) Normal beat. (C) LBBB beat. (D) Nodal escape beat. (E) Nodal
premature beat. (F) Fusion of ventricular and normal beat. (G) Atrial escape beat. (H) Ventricular escape beat. (I) Atrial premature beat. (J) Aberrated atrial premature
beat. (K) Premature ventricular contraction. (L) Premature or ectopic supraventricular beat. March 2022 | Volume 9 | Article 857019 Frontiers in Cardiovascular Medicine | www.frontiersin.org 10 Beat-Level Interpretation of Intra-Patient Paradigm Based on Object Detection Kang et al. 0.02 and divided by 10 at the 16th and 22nd epoch. The weight
decay and momentum are set to 0.0001 and 0.9, respectively. implicitly encodes contextual information about the class and
appearance to detect foreground objects more accurately, as well
as directly optimizes the detection performance end-to-end. The optimal model with the highest mAP@0.5 is obtained by
using 5-fold cross-validation during training. Table 3 shows its
prediction accuracy of each category on the testing set. y p
p
This work constructs an object detection framework for
heartbeat images based on Cascade RCNN, which consists of a
series of cascading detection networks. Each detection network
is trained on positive and negative samples based on different
IoU thresholds. The output of the former network serves as the
input to the latter, which is a stage-by-stage training method. The detector of each stage focuses on detecting the proposals
whose IoU are in a certain range. The detection effect gets better
and better since the output IoU is generally larger than the
input IoU. We take a part of the heartbeat images as samples;
Table 5 shows the detection accuracy of Faster RCNN (51) and
Cascade RCNN on the four categories recommended by AAMI. Obviously, Cascade RCNN is more effective, whose mAP@0.5
value is about 2 percent higher than Faster RCNN. It is worth noting that the method in this work is not limited
to the Cascade RCNN object detector, but applicable to other
two-stage detectors or one-stage detectors. We could choose the
appropriate detector as the basic model according to your needs. The Effectiveness of Automatic Location
Annotation On the one hand, the morphology of ECG varies from person
to person, even the same individual at different times, and the
signals are easily disturbed by noises, which resulting in the
difficulty of QRS complex detection. On the other hand, the QRS
complex detection process is separate, so it is hard to achieve an
end-to-end overall optimization through model training. It takes
much time and effort, which is not beneficial for real-time testing. As we all known that the annotations play an important role in
an object detection task, but only about 200–300 images can be
processed per hour if we manually annotate images one by one
like the similar work (51, 52). Obviously, the high human cost
will discourage users and hardly expand to larger datasets, which
deeply reduce the value of this kind of method. g
This work avoids independent QRS complexes detection,
which decouples the high dependence of feature extraction and
heartbeats classification on detection accuracy. A core strategy in this work is to automatically annotate
the QRS complex position of the heartbeat images, which
avoids the extremely time-consuming manual labeling process. Thousands of images can be annotated in just a few minutes, so
instead of fixed lengths like one beat length (51) or 10 s (52),
we can easily get the signal sequences of different lengths, as
shown in Figure 4, which is beneficial for testing samples of
unknown length. Overall Classification Performance In order to take the position information as an object learned
by the model, this work converts one-dimensional signals
into two-dimensional images whose QRS complex position is
annotated by automatic annotation algorithm. The positioning
and classification tasks are unified in this work. Except for the improved classification accuracy as described
in section Overall Classification Performance, the annotation is
also essential for reducing the complexity during the inference
stage. In the testing mode of traditional methods, as shown in
Figure 1A, the preprocessed signals still need to perform the
same process including QRS detection and feature extraction
as in the training mode. The training stage does not facilitate
the inference stage. The proposed method spends some labeling
costs during the training stage, while in the testing mode, as
shown in Figure 1B, the preprocessed signals need only be fed
into the pretrained model as images. Figure 9 visualizes the
prediction results for different categories of heartbeats; the model
could output both the position of the QRS complex in the
form of a rectangular box and the heartbeat category with a
confidence score. This work focuses on the interpretation of beat-level. Especially for an ECG signal containing multiple types of
heartbeats such as bigeminy and trigeminy, we can get a detailed
category of each beat for further analysis, rather than only the
existence of abnormality by just classifying a signal over a long
period of time. In general, this work shows good performance in the beat-
level classification of intra-patient paradigm. Table 4 shows a
comparison with previous work. It can be seen that our method
achieves the best results in both average classification accuracy
and specificity. The sensitivity is slightly lower than some work
which has fewer categories of interpretation. Obviously, our
method is more advantageous in the number of categories. CONCLUSION AND FUTURE
PERSPECTIVE In this work, we propose a beat-level interpretation method based
on object detection. We use a convolutional neural network as a
feature extractor for two-dimensional heartbeat images, without Frontiers in Cardiovascular Medicine | www.frontiersin.org March 2022 | Volume 9 | Article 857019 11 Kang et al. Beat-Level Interpretation of Intra-Patient Paradigm Based on Object Detection FUNDING This work was supported by the National Natural Science
Foundation of China (Grant 62022047, Grant 61874065, Grant
U20A20168, and Grant 51861145202), Beijing Natural Science
Foundation under Grant M22020, the National Key R&D
Program under Grant 2016YFA0200400, in part by Fok Ying-
Tong Education Foundation under Grant 171051, in part by
Beijing National Research Center for Information Science and
Technology Youth Innovation Fund (BNR2021RC01007), in part
by State Key Laboratory of New Ceramic and Fine Processing
Tsinghua University (No. KF202109), in part by the Research
Fund from Beijing Innovation Center for Future Chip and the
Independent Research Program of Tsinghua University under
Grant 20193080047, and Tsinghua-Foshan Innovation Special
Fund (2021THFS0217). DATA AVAILABILITY STATEMENT Publicly available datasets were analyzed in this study. This data
can be found here: physionet.org/content/mitdb/1.0.0/. AUTHOR CONTRIBUTIONS complex manual design to extract features. Most importantly,
this work abandons the previous mode of separate QRS complex
detection and heartbeats classification, the ground truth of QRS
complex is marked by automatic annotation algorithm, which
is also regarded as the object, the model can learn like category
information. The classification and regression branches of the
object detector unify the localization and classification tasks,
achieving an end-to-end optimization as well as decoupling
the high dependence on the R-peak detection. We evaluate the
performance on the MIT-BIH database, our method is superior
to most advanced research even if the number of categories is
as many as 12. The average accuracy is 99.60%, the average
sensitivity is 97.56%, and the average specificity is 99.78%. In addition, since the independent and time-consuming QRS
complex detection process is abandoned during the inference
stage, our method is expected to be adopted in real-time
monitoring systems to bring convenience to patients with cardiac
abnormalities in the future. Of course, the MIT-BIH database
contains too few patients to support the classification of inter-
patient paradigm (53); the method in this work can be extended
to the inter-patient paradigm when the beat-level annotations of
more patients are obtained in the future. MK and X-FW contributed to conception and design of the
study. MK wrote the codes and ran experiments on the relevant
dataset and wrote the first draft of the manuscript. X-FW
performed the statistical analysis. JX help to optimize the
detection network. HT and T-LR supervised the whole work. X-FW, JX, HT, and T-LR refined the manuscript. All authors
contributed to manuscript revision, read, and approved the
submitted version. REFERENCES 10. Karpagachelvi
S,
Arthanari
M,
Sivakumar
M. Classification
of
electrocardiogram
signals
with
support
vector
machines
and
extreme
learning
machine. Neural
Comput
Appl. (2012)
21:1331–9. doi: 10.1007/s00521-011-0572-z 1. Pan J, Tompkins WJ. A real-time QRS detection algorithm. IEEE
Trans
Biomed
Eng. (2007)
32:230–6. doi:
10.1109/TBME.1985. 325532 1. Pan J, Tompkins WJ. A real-time QRS detection algorithm. IEEE
Trans
Biomed
Eng. (2007)
32:230–6. doi:
10.1109/TBME.1985. 325532 11. Manu T, Manab KD, Samit A. Automatic ECG arrhythmia classification using
dual tree complex wavelet based features. IEEE Int J Electron Commun. (2015)
69:715–21. doi: 10.1016/j.aeue.2014.12.013 2. Yeh YC, Wang WJ. QRS complexes detection for ECG signal: the
difference operation method. Comp Methods Progr Biomed. (2008) 91:245–
54. doi: 10.1016/j.cmpb.2008.04.006 2. Yeh YC, Wang WJ. QRS complexes detection for ECG signal: the
difference operation method. Comp Methods Progr Biomed. (2008) 91:245–
54. doi: 10.1016/j.cmpb.2008.04.006 12. Raj
S,
Ray
KC. ECG
signal
analysis
using
DCT-based
DOST
and
PSO
optimized
SVM. IEEE
Trans
Inst
Meas. (2017)
66:470–8. doi: 10.1109/TIM.2016.2642758 3. Li CW, Zheng CX, Tai CF. Detection of ECG characteristic points
using wavelet transforms. IEEE Trans Biomed Eng. (1995) 42:21–28. doi: 10.1109/10.362922 3. Li CW, Zheng CX, Tai CF. Detection of ECG characteristic points
using wavelet transforms. IEEE Trans Biomed Eng. (1995) 42:21–28. doi: 10.1109/10.362922 13. Li HQ, Liang H, Miao CJ, Cao L, Feng XL, Tang CX, et al. Novel ECG signal
classification based on KICA nonlinear feature extraction. IEEE Circuits Syst
Signal Process. (2016) 35:1187–97. doi: 10.1007/s00034-015-0108-3 4. Martínez JP, Almeida R, Olmos S, Rocha AP, Laguna P. A wavelet-based ECG
delineator: evaluation on standard databases. IEEE Trans Biomed Eng. (2004)
51:570. doi: 10.1109/TBME.2003.821031 4. Martínez JP, Almeida R, Olmos S, Rocha AP, Laguna P. A wavelet-based ECG
delineator: evaluation on standard databases. IEEE Trans Biomed Eng. (2004)
51:570. doi: 10.1109/TBME.2003.821031 14. Hu JI. Research on Key Technologies for Automatic Analysis of ECG Signals. Changsha: National University of Defense Technology (2006). 5. Manikandan MS, Soman KP. A novel method for detecting R-peaks in
electrocardiogram (ECG) signal. Biomed Signal Process Control. (2012) 7:118–
28. doi: 10.1016/j.bspc.2011.03.004 15. Song MH, Lee J, Cho SP, Lee KJ, Yoo SK. Support vector machine
based arrhythmia classification using reduced features. In: International
Journal
of
Control
Automation
and
Systems. (2005)
p. 571-9. doi: 10.1007/s00170-004-2187-3 6. Sukanta
S,
Santanu
S,
Bhupen
K,
Suresh
B. Multiresolution
wavelet
transform
based
feature
extraction
and
ECG
classification
to
detect
cardiac
abnormalities. IEEE
Meas. (2017)
108:55–
66. REFERENCES One-dimensional
CNN
approach
for
ECG
arrhythmia
analysis
in
fog-cloud
environments. IEEE
Access. (2021)
9:103513–
23. doi: 10.1109/ACCESS.2021.3097751 42. Niu
JH,
Tang
YQ,
Sun
ZY,
Zhang
WS. Inter-patient
ECG
classification
with
symbolic
representations
and
multi-perspective
convolutional neural networks. IEEE J Biomed Health Informat. (2019)
24:1321–32. doi: 10.1109/JBHI.2019.2942938 23. Maweu B, Dakshit S, Shamsuddin R, Prabhakaran B. CEFEs: a CNN
explainable
framework
for
ECG
signals. Artif
Intell
Med. (2021)
115:102059. doi: 10.1016/j.artmed.2021.102059 43. Houssein
EH,
Ibrahim
E,
Neggaz
N,
Hassaballah
M,
Wazery
Y. An
efficient
ECG
arrhythmia
classification
method
based
on
Manta
ray
foraging
optimization. Expert
Syst
Appl. (2021)
181:115131. doi: 10.1016/j.eswa.2021.115131 24. Li XL, Panicker RC, CardiffB, John D. Multistage pruning of CNN based
ECG classifiers for edge devices. arXiv [Preprint]. (2021) arXiv: 2109.00516. doi: 10.48550/arXiv.2109.00516 44. Naz M, Shah JH, Khan MA, Sharif M, Raza M, Damasevicius R. From ECG
signals to images: a transformation based approach for deep learning. PeerJ
Comp Sci. (2021) 7:e386. doi: 10.7717/peerj-cs.386 25. Acharya UR, Oh SL, Hagiwara Y, Tan HJ, Adam M, Gertych A, et al. A deep
convolutional neural network model to classify heartbeats. Comput Biol Med. (2017) 89:389–96. doi: 10.1016/j.compbiomed.2017.08.022 45. Ren SQ, He KM, Girshick R, Sun J. Faster R-CNN: towards real-time object
detection with region proposal networks. IEEE Trans Pattern Anal Mach
Intell. (2017) 39:1137–49. doi: 10.1109/TPAMI.2016.2577031 26. Chauhan S, Vig L. Anomaly detection in ECG time signals via deep long short-
term memory networks. In: IEEE International Conference on Data Science &
Advanced Analytics. Paris: IEEE (2015). 46. American National Standard for Testing and Reporting Performance Results
of Cardiac Rhythm and ST Segment Measurement Algorithms. (1998)
AAMI/ANSI Standard EC57:1998. 27. Eltrass AS, Tayel MB, Ammar AI. A new automated CNN deep learning
approach for identification of ECG congestive heart failure and arrhythmia
using constant-Q non-stationary Gabor transform. Biomed Signal Process
Control. (2021) 65:102326. doi: 10.1016/j.bspc.2020.102326 47. Lin TY, Dollár P, Girshick R, He KM, Hariharan B, Belongie S. Feature
pyramid networks for object detection. In: Proceedings of the IEEE Conference
on Computer Vision and Pattern Recognition. IEEE (2017) p. 2117–25. doi: 10.1109/CVPR.2017.106 28. Warrick
P,
Homsi
MN. Cardiac
arrhythmia
detection
from
ECG
combining convolutional and long short-term memory networks. In:
2017 Computing in Cardiology (CinC). Rennes: IEEE (2017) p. 1–4. doi: 10.22489/CinC.2017.161-460 48. He KM, Zhang XY, Ren SQ, Sun J. Deep residual learning for image
recognition. Comp Vis Pattern Recogn. (2016) 2:6.doi: 10.1109/CVPR.2016.90 29. REFERENCES doi: 10.1016/j.measurement.2017.05.022 6. Sukanta
S,
Santanu
S,
Bhupen
K,
Suresh
B. Multiresolution
wavelet
transform
based
feature
extraction
and
ECG
classification
to
detect
cardiac
abnormalities. IEEE
Meas. (2017)
108:55–
66. doi: 10.1016/j.measurement.2017.05.022 16. Melgani F, Bazi Y. Classification of electrocardiogram signals with support
vector machines and particle swarm optimization. In: IEEE Transactions
on Information Technology in Biomedicine, Vol. 12. IEEE (2008) p. 667–77. doi: 10.1109/TITB.2008.923147 7. Patro
KK,
Kumar
PR. Effective
feature
extraction
of
ECG
for
biometric
application. IEEE
Procedia
Comput
Sci. (2017)
115:296–306. doi: 10.1016/j.procs.2017.09.138 17. Martis RJ, Acharya UR, Mandana KM, Ray AK, Chakraborty C. Cardiac
decision making using higher order spectra. Biomed Signal Process Control. (2013) 8:193–203. doi: 10.1016/j.bspc.2012.08.004 8. Elhaj FA, Salim N, Harris AR, Swee TT, Ahmed T. Arrhythmia recognition
and
classification
using
combined
linear
and
nonlinear
features
of
ECG signals. IEEE Comput Methods Progr Biomed. (2016) 127:52–
63. doi: 10.1016/j.cmpb.2015.12.024 18. Ganeshkumar R, Dr. Kumaraswamy YS. Investigating cardiac arrhythmia
in ECG using random forest classification. Int J Comp Appl. (2012) 37:31–
4. doi: 10.5120/4599-6557 9. Liu CY, Zhang XY, Zhao LN, Liu FF, Chen XW, Yao YJ, et al. Signal quality assessment and lightweight QRS detection for wearable
ECG SmartVest system. IEEE Internet of Things J. (2019) 6:1363–
74. doi: 10.1109/JIOT.2018.2844090 19. Park J, Lee K, Kang K. Arrhythmia detection from heartbeat using k-nearest
neighbor classifier. In: IEEE International Conference on Bioinformatics and
Biomedicine. Shanghai: IEEE Computer Society (2013). March 2022 | Volume 9 | Article 857019 Frontiers in Cardiovascular Medicine | www.frontiersin.org 12 Beat-Level Interpretation of Intra-Patient Paradigm Based on Object Detection Kang et al. 40. Wang JB. A deep learning approach for atrial fibrillation signals classification
based on convolutional and modified Elman neural network. Future Gen
Comp Syst. (2020) 102:670–9. doi: 10.1016/j.future.2019.09.012 20. Jun TJ, Park HJ, Yoo H, Kim YH, Kim D. GPU based cloud system for
high-performance arrhythmia detection with parallel k-NN algorithm[C]. In:
Engineering in Medicine and Biology Society. Miami, FL: IEEE (2016). 41. Chen C, Hua ZC, Zhang RQ, Liu GY, Wen WH. Automated arrhythmia
classification based on a combination network of CNN and LSTM. Biomed Signal Process Control. (2020) 57:101819. doi: 10.1016/j.bspc.2019. 101819 21. Kiranyaz S, Ince T, Gabbouj M. Real-time patient-specific ECG classification
by 1-D convolutional neural networks. IEEE Trans Biomed Eng. (2016)
63:664–75. doi: 10.1109/TBME.2015.2468589 22. Cheikhrouhou
O,
Mahmud
R,
Zouari
R,
Ibrahim
M,
Zaguia
A,
Gia
TN. REFERENCES Shu
LO,
Eddie
YK,
Tan
RS,
Acharya
UR. Automated
diagnosis
of
arrhythmia
using
combination
of
CNN
and
LSTM
techniques
with
variable
length
heart
beats. Comput. Biol. Med. (2018)
102:278–87. doi: 10.1016/j.compbiomed.2018.06.002 49. He K, Gkioxari G, Dollár P, Girshick R. Mask R-CNN. In: Proceedings of
the IEEE international conference on computer vision. Venice, Italy, (2017)
2961–2969. doi: 10.1109/ICCV.2017.322 50. Chen K, Wang JQ, Pang JM, Cao YH, Xiong Y, Li XX, et al. Mmdetection:
Open mmlab detection toolbox and benchmark. arXiv [Preprint]. (2019)
arXiv: 1906.07155. doi: 10.48550/arXiv.1906.07155 30. Wong D, Li YF, John D, Ho WK, Heng CH. Resource and energy efficient
implementation of ECG classifier using binarized CNN for edge AI devices. In:
2021 IEEE International Symposium on Circuits and Systems (ISCAS). Daegu:
IEEE (2021). 51. Ji YS, Zhang S, Xiao WD. Electrocardiogram classification based on
faster
regions
with
convolutional
neural
network. Sensors. (2019)
19:2558. doi: 10.3390/s19112558 31. Yao GL, Mao XB, Li N, Xu HX, Xu XY, Jiao Y, et al. Interpretation of
electrocardiogram heartbeat by CNN and GRU. Comput Math Methods Med. (2021) 2021:6534942. doi: 10.1155/2021/6534942 52. Hwang WH, Jeong CH, Hwang DH, Jo YC. Automatic detection of
arrhythmias using a YOLO-based network with long-duration ECG signals. Eng Proc. (2020) 2:84. doi: 10.3390/ecsa-7-08229 32. Sarin E, Taneja S, Vividha, Nagrath P. Analyzing and Comparing MLP, CNN,
and LSTM for Classification of Heart Arrhythmia Using ECG Scans. In:
Robotic Technologies in Biomedical and Healthcare Engineering. CRC Press
(2021) p. 87–115. doi: 10.1201/9781003112273-5 53. Mondéjar-Guerra V, Novo J, Rouco J, Gonzalez M, Ortega M. Heartbeat
classification fusing temporal and morphological information of ECGs
via ensemble of classifiers. Biomed Signal Process Control. (2018) 47:41–
8. doi: 10.1016/j.bspc.2018.08.007 33. Saxena
SC,
Kumar
V,
Hamde
ST. QRS
detection
using
new
wavelets. J
Med
Eng
Technol. (2002)
26:7. doi:
10.1080/03091900
110096038 Conflict of Interest: JX was employed by Ping An Technology (Shenzhen) Co. Ltd. 34. Mallat
S,
Hwang
WL. Singularity
detection
and
processing
with
wavelets. IEEE Trans Inf Theory. (1992) 38:P.617–43. doi: 10.1109/18. 119727 The remaining authors declare that the research was conducted in the absence of
any commercial or financial relationships that could be construed as a potential
conflict of interest. 35. Cai Z, Vasconcelos N. Cascade r-cnn: delving into high quality object
detection[C]. In: Proceedings of the IEEE Conference on Computer Vision and
Pattern Recognition. Salt Lake City, UT: IEEE (2018) p. 6154–62. Publisher’s Note: All claims expressed in this article are solely those of the authors
and do not necessarily represent those of their affiliated organizations, or those of
the publisher, the editors and the reviewers. Any product that may be evaluated in
this article, or claim that may be made by its manufacturer, is not guaranteed or
endorsed by the publisher. 36. Zhou
S,
Tan
B. Electrocardiogram
soft
computing
using
hybrid
deep
learning
CNN-ELM. Appl
Soft
Comput. (2019)
86:105778. doi: 10.1016/j.asoc.2019.105778 37. Hou B, Yang JY, Wang P, Yan RQ. LSTM-based auto-encoder model for
ECG arrhythmias classification. IEEE Trans Instrum Meas. (2019) 69:1232–
40. doi: 10.1109/TIM.2019.2910342 Copyright © 2022 Kang, Wang, Xiao, Tian and Ren. This is an open-access article
distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the
original author(s) and the copyright owner(s) are credited and that the original
publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these
terms. 38. Wan XK, Jin ZY, Wu HB, Liu JJ, Zhu BR, Xie HG. Heartbeat
classification algorithm based on one-dimensional convolution neural
network. J Mech Med Biol. (2020) 20:2050046. doi: 10.1142/S0219519420
500463 39. Ullah A, Anwar S, Bilal M, Mehmood RM. Classification of arrhythmia by
using deep learning with 2-D ECG spectral image representation. Remote Sens. (2020) 12:1685. doi: 10.3390/rs12101685 March 2022 | Volume 9 | Article 857019 Frontiers in Cardiovascular Medicine | www.frontiersin.org 13
|
https://openalex.org/W4323075395
|
https://journal.formosapublisher.org/index.php/jpmf/article/download/2106/1864
|
Indonesian
| null |
Prevention of the Spread of the COVID-19 Virus with an Online to Offline Shopping System with an Independent Self-Service Concept using a Miniature Solar-Powered Mobile Market
|
Jurnal Pengabdian Masyarakat Formosa
| 2,023
|
cc-by
| 2,537
|
Jurnal Pengabdian Masyarakat Formosa (JPMF)
Vol. 2, No. 1, 2023 21- 28 Prevention of the Spread of the COVID-19 Virus with an Online
to Offline Shopping System with an Independent Self-Service
Concept using a Miniature Solar-Powered Mobile Market Anantha Zakharia1, Irvan Hermala2, Putie Maharani3*
1, 3Universitas Bina Sarana Informatika , 2Universitas Mercu Buana
Corresponding Author: Putie Maharani. putie.pmb@bsi.ac.id A R T I C L E I N F O A R T I C L E I N F O
Keywords: Covid-19, Corona
Virus, Economic Recovery A R T I C L E I N F O
Keywords: Covid-19, Corona
Virus, Economic Recovery A B S T R A C T The participation of universities is highly
expected in assisting economic recovery, so that
economic turnover activities can continue
without increasing the number of Covid-19
sufferers. In this case, universities provide
solutions in the field of renewable energy as a
source of electricity to support the efficiency of
the prototypes produced. This activity was
carried out by students and lecturers as a form
of community service. Where the concept
offered is online to offline to maintain shopping
loyalty. Consumers can make transactions by
selecting product quality, tasting the product,
and tasting the product first and they do it in
front of their house in the form of a solar-
powered mobile market. The method used in
the data collection process: the Preparation
Stage, the Group Discussion Stage, the Capacity
Building Implementation Stage: the Research
Stage, the Data Analysis Stage. The conclusion
from this study is that the community is
enthusiastic, especially those living in housing
because of the online to offline system (online
ordering system, goods come offline, consumers
can choose). For vehicles, it can save fuel
because it uses a solar power engine. Received : 02 December
Revised : 05 January
Accepted: 10 February ©2023
Zakharia,
Hermalam,
Maharani This is an open-access
article distributed under the terms of
the Creative Commons Atribusi 4.0
Internasional. (
DOI: https://doi.org/10.55927/jpmf.v2i1.2106
ISSN-E: 2829-6486
https://journal.formosapublisher.org/index.php/jpmf 21 Zakharia, Hermalam, Maharani Pencegahan Penyebaran Virus COVID-19 dengan Sistem Belanja
Online to Offline Berkonsep Swalayan Mandiri menggunakan
Miniatur Pasar Keliling Bertenaga Matahari Anantha Zakharia1, Irvan Hermala2, Putie Maharani3*
1, 3Universitas Bina Sarana Informatika , 2Universitas Mercu Buana
Corresponding Author: Putie Maharani. putie.pmb@bsi.ac.id A R T I C L E I N F O
Kata Kunci: Covid-19, Virus
Corona, Pemulihan Ekonomi A R T I C L E I N F O
Kata Kunci: Covid-19, Virus
Corona, Pemulihan Ekonomi A B S T R A K
Peran serta dari perguruan tinggi sangat
diharapkan
dalam
membantu
pemulihan
ekonomi,
sehingga
kegiatan
perputaran
ekonomi
tetap
berjalan
tanpa
adanya
penambahan jumlah penderita Covid-19. Dalam
hal ini perguruan tinggi memberikan solusi di
bidang energi terbarukan sebagai sumber listrik
untuk mendukung efisiensi dari purwarupa
yang dihasilkan. Kegiatan ini dilakukan oleh
mahasiswa
dan
dosen
sebagai
bentuk
pengabdian masyarakat. Dimana konsep yang
ditawarkan adalah online to offline untuk
menjaga loyalitas belanja. Konsumen bisa
melakukan transaksi dengan memilih kualitas
produk, merasakan produk, dan meraba produk
terlebih dahulu dan mereka melakukannya
depan rumah mereka dalam bentuk pasar
keliling
bertenaga
surya. Metode
yang
digunakan dalam proses pengumpulan data:
Tahap
Persiapan,
Tahap
Penyelenggaraan
Diskusi
Kelompok,
Tahap
Pelaksanaan
Peningkatan Capacity Building: Tahap Penelitian,
Tahap
Analisis
Data. Kesimpulan
dari
penelitian ini adalah masyarakat antusias,
terutama yang bermukim di perumahan karena
dengan sistem online to offline (sistem pesan
online, barang datang offline, konsumen bisa
memilih). Untuk kendaraan dapat mengirit
bahan bakar karena menggunakan mesin tenaga
surya. Received : 02 Desember
Revised : 05 Januari
Accepted: 10 Februari Received : 02 Desember
Revised : 05 Januari
Accepted: 10 Februari ©2023
Zakharia,
Hermalam,
Maharani This is an open-access
article distributed under the terms of
the Creative Commons Atribusi 4.0
Internasional. 22 22 Jurnal Pengabdian Masyarakat Formosa (JPMF)
Vol. 2 No 1, 2023: 21 - 28 PENDAHULUAN PENDAHULUAN Pandemi COVID-19 telah menghancurkan perekonomian di Indonesia
sejak bulan Maret 2020, dimana semua daerah menerapkan Pembatasan Sosial
Berskala Besar (PSBB) dan Pembatasan Sosial Berskala Mikro (PSBM). Hal ini
menyebabkan masyarakat mengurangi kegiatan berbelanja dan pembatasan
aktivitas bisnis oleh pemerintah setempat. Perguruan tinggi dalam perannya sebagai lembaga pendidikan formal di
harapkan dapat memberikan solusi dalam permasalahan tersebut. Dalam
pengabdian masyarakat yang di lakukan oleh Universitas Bina Sarana
Informatika berkolaborasi dengan Universitas Mercu Buana mengusung tema
online to offline untuk mencegah penyebaran virus COVID-19 dengan
menggunakan kendaraan tenaga surya sebagai energi terbarukan dengan
sistem door to door berdasarkan pesanan. Ide ini muncul ketika adanya sistem pembelanjaan online dimana
konsumen tidak dapat memilih kualitas produk, merasakan produk, dan
meraba produk sebelum transaksi dilakukan. Sehingga terjadi kesenjangan
antara harapan tentang produk yang diinginkan oleh konsumen dengan
realisasi produk ketika produk tersebut sampai di tangan konsumen. Dengan
adanya ide ini, selain sebagai alternatif solusi, juga diharapkan adanya
pergerakan ekonomi yang signifikan pada dunia usaha. Seperti munculnya
startup-startup
baru
di
dunia
industri. Sehingga
dapat
mengurangi
dampak/efek dari pandemi itu sendiri. Dalam hal ini pengurangan tenaga kerja
(PHK). Pelaksanaan dan Metode Pelaksanaan dan Metode
Gambar 1 Alur Roadmap dan Desain Program Terhadap Peluang Cipta Gambar 1 Alur Roadmap dan Desain Program Terhadap Peluang Cipta 23 Zakharia, Hermalam, Maharani y p
p
2. Metode Penentuan Peralatan Penelitian Peralatan yang akan digunakan adalah dua unit kendaraan dari PT
Nadelsa Kreasi Indonesia yang akan dimodifikasi menjadi miniatur pasar
keliling. Jadi, teknologi solar cell dari Universitas Mercu Buana akan
dikombinasikan pada kendaraan tersebut. 3. Metode Penentuan Lokasi Penelitian Menurut informasi yang didapatkan dari Satgas Covid-19 Kota Bogor
bahwa penduduk yang terinfeksi sering terjadi di daerah komplek perumahan. Oleh karena itu, lokasi aktivitas penelitian akan difokuskan pada komplek
perumahan yang ada di wilayah Kota Bogor. Sebagai pelaksana pemilihan
lokasi ini adalah pihak dari Universitas Bina Sarana Informatika yang mana
lokasi kampus juga berada di wilayah Kota Bogor dan PT Nadelsa Kreasi
Indonesia yang memiliki aset berlokasi di tengah Kota Bogor untuk bisa
dimanfaatkan sebagai mini power plant miniatur pasar keliling. . 1. Metode Penentuan Bahan Penelitian 1. Metode Penentuan Bahan Penelitian Pemilihan bahan-bahan yang akan digunakan selama kegiatan sebagai
penyedia consumer good yang akan dikonsumsi oleh konsumen sehingga
tercipta loyalitas konsumen. Dimana produk akan dipajang pada miniatur
pasar keliling seperti sayuran, buah-buahan, sembako, bumbu masakan,
makanan olahan, berbagai jenis daging, frozen food, pembersih tubuh, keperluan
bayi, pembersih rumah, bahan pembuat kue, minuman dll. B. Tahap Penyelenggaraan Diskusi Kelompok
1. Metode Pemilihan Peserta Peserta yang dipilih adalah warga komplek perumahan yang ada di
Kota Bogor dengan target jumlah peserta adalah 2.000 orang yang tersebar di
beberapa perumahan Kota Bogor. Maksud dari pemilihan peserta karena
adanya korelasi pada penelitian sebelumnya. Acuan pemilihan peserta yang
nantinya digunakan sebagai objek penelitian adalah: Hasil penelitian Dwi Martiyanti Fakultas Ekonomi dan Bisnis
Universitas Mulawarman tentang Perbedaan Gender Mengenai Perceived
Risk pada Pembelian Online sebagai acuan untuk mengetahui tentang
resiko yang dirasakan setiap gender ketika belanja. Data Badan Pusat Statistik Kota Bogor tahun 2021 untuk mendapatkan
info tentang populasi penduduk berjenis kelamin wanita di Kota Bogor. Buku terbitan Rineka Cipta karangan Suharsimi Arikunto tahun 2002
yang berjudul Prosedur Penelitian Suatu Pendekatan Praktek sebagai
acuan pengambilan jumlah sampel minimum. Buku terbitan Rineka Cipta karangan Suharsimi Arikunto tahun 2002
yang berjudul Prosedur Penelitian Suatu Pendekatan Praktek sebagai
acuan pengambilan jumlah sampel minimum. p
g
j
p
2. Metode Penyampaian Diskusi dan Materi Diskusi p
g
j
p
2. Metode Penyampaian Diskusi dan Materi Diskusi Diskusi yang akan dilakukan nantinya tetap memperhatikan protokol
kesehatan. Oleh karena itu, diskusi akan dilakukan secara bertahap terhadap
2.000. Peserta dibatasi masing-masing tahap untuk mempermudah penerapan
social distancing. Mereka akan dilengkapi alat pelindung diri seperti masker dan
hand sanitizer. Pemberian materi pada diskusi akan melibatkan narasumber untuk
memberikan edukasi seputar bahaya belanja offline selama pandemi. Selain itu, 24 Jurnal Pengabdian Masyarakat Formosa (JPMF)
Vol. 2 No 1, 2023: 21 - 28 Jurnal Pengabdian Masyarakat Formosa (JPMF)
Vol. 2 No 1, 2023: 21 - 28 akan dibuka sesi tanya jawab untuk menggali permasalah peserta secara
deskriptif tentang kegiatan belanja yang sudah dilakukan selama pandemi. Melalui kegiatan ini juga akan disosialisasikan konsep belanja swalayan
mandiri online to offline dan prototype miniatur pasar keliling kepada para
peserta. Sebagai pelaksana pada tahap ini adalah pihak dari Universitas Bina
Sarana Informatika dan pihak dari PT Nadelsa Kreasi Indonesia. C. Tahap Pelaksanaan Peningkatan Capacity Building Capacity building dalam hal ini dilakukan agar mendapat arahan untuk
pengabdian/penelitian ke depan agar dapat dikomersialkan. Oleh karena itu,
dibutuhkan mentor yang mempunyai wawasan dalam hal pengetahuan,
pengalaman, dan juga kemampuan untuk membantu memvalidasi ide pada
pengabdian ini agar mudah diterima oleh pasar. 1. Metode Penguatan Promosi Penguatan promosi ditujukan sebagai kegiatan branding selama masa
pengabdian supaya objek peneliti mau menggunakan sistem dan purwarupa
yang telah disosialisasikan. Oleh karena itu, pada aktivitas ini akan melibatkan
tenaga ahli berpengalaman di bidang branding. Sebagai pemantau pada metode
ini nantinya akan dilakukan oleh tim dari PT Nadelsa Kreasi Indonesia. 2
d S
i y
2. Metode Survei y
2. Metode Survei Survei dilakukan kepada 2.000 responden peserta bersamaan dengan
aktivitas Focus Group Discussion (FGD). Dalam pengabdian diterapkan
penggunaan sistem IoT untuk mempermudah pelaksanaanya. Dengan
demikian, akan melibatkan tenaga ahli di bidang teknologi IoT untuk
mendukung pelaksanaan jalannya pengabdian tersebut. Yang akan dilakukan
oleh pihak Universitas Bina Sarana Informatika. 1. Metode Pengolahan Data Pengolahan data tentang loyalitas konsumen pada sistem swalayan
mandiri online to offline akan menggunakan uji validitas, uji reliabilitas, uji
normalitas, uji multikolinearitas, dan uji heteroskedastisitas. Selanjutnya, untuk
transformasi data dilakukan dengan mengkonversi skala kuesioner yang
semula berskala ordinal menjadi interval. Sedangkan untuk pengolahan data
hasil pengamatan prototype miniatur pasar keliling bertenaga surya akan
menggunakan uji independent T-test. Pada kasus ini akan membandingkan
biaya operasional pada kedua perlakuan purwarupa pasar keliling tersebut. 2. Metode Penyajian Data Penyajian data penelitian akan dilakukan dalam tiga macam dengan
rincian sebagai berikut: Penyajian data penelitian akan dilakukan dalam tiga macam denga
rincian sebagai berikut: Narasi : penyajian data hasil penelitian akan dijabarkan dalam bentuk
kalimat. Tabel : penyajian data hasil penelitian berupa kumpulan angka-angka
yang disusun menurut kategori tertentu. Grafik : penyajian data hasil penelitian dalam bentuk infografis. Gambar : penyajian data hasil penelitian dalam bentuk foto. Gambar : penyajian data hasil penelitian dalam bentuk foto. Diatas adalah metode/cara yang dilakukan dalam pengabdian ini. iatas adalah metode/cara yang dilakukan dalam pengabdian ini. p
3. Metode Pengamatan Pelaksanaan dilakukan oleh pihak dari Universitas Bina sarana
Informatika dalam hal pengamatan loyalitas objek penelitian menggunakan
sistem swalayan mandiri online to offline. Beberapa hal yang akan diamati
adalah sebagai berikut: Kelengkapan produk terhadap loyalitas pelanggan Waktu tempuh pengiriman terhadap loyalitas pelanggan Harga produk terhadap loyalitas pelanggan Kualitas produk terhadap loyalitas pelanggan Recording data terhadap metode pengamatan di atas akan menggunakan
sistem
aplikasi
berbasis
internet. Pengamatan
juga
dilakukan
pada
purwarupa/prototype miniatur pasar keliling yang dilaksanakan oleh pihak dari
Universitas Mercu Buana. Perlakuan yang diberikan adalah mobil yang
dimodifikasi menjadi miniatur pasar keliling yang disematkan baterai panel
surya. Parameter yang diamati adalah biaya operasional yang dikeluarkan oleh
miniatur pasar keliling baik ketika beroperasi maupun ketika me-recharge
kembali energi surya tersebut. 25 Zakharia, Hermalam, Maharani HASIL DAN PEMBAHASAN Hasil dari pengabdian ini dituangkan dalam bentuk narasi deskriptif,
karena pada pelaksanaannya baru sampai tahap B yaitu Penyelenggaraan
Diskusi Kelompok. Dari hasil diskusi di peroleh kesimpilan bahwa masyarakat
antusias dengan Program ini, dimana penerima manfaat adalah warga komplek
perumahan yang ada di Kota Bogor. Mereka bisa melakukan kegiatan belanja
seperti di tempat perbelanjaan umum dengan minim potensi terinfeksi Virus
Covid-19 karena menggunakan sistem swalayan mandiri online to offline di
tempat tinggal masing-masing. p
gg
g
g
Kelompok penerima manfaat kedua adalah sektor Usaha Kecil dan
Menengah (UMKM) yang terdampak selama pandemi berlangsung. Angkatan
kerja baru yang merupakan lulusan dari universitas juga bisa menerima
manfaat dari pengabdian ini untuk berwirausaha dengan jumlah 139,81 juta
(Data BPS, 2021). Perlu diketahui, bahwa pelaku UMKM dan para angkatan
kerja baru bisa memanfaatkan purwarupa pasar keliling untuk menjalankan
kegiatan usahanya di tengah pandemi dengan cara menjemput bola. Selain itu,
sejumlah
mahasiswa
di
bidang
ekonomi
sebanyak
1,7
juta
(Data
Kemenristekdikti, 2019) bisa belajar tentang tatanan seputar sharing economic
model
terbaru
dilapangan. Selain
mahasiswa
ekonomi
mahasiswa
teknik&Elektro dapat mengembangkan sumber energi listrik tenaga Surya. Sedangkan penerima manfaat terakhir adalah para stakeholder pengabdian
seperti mitra PT Nadelsa Kreasi Indonesia, dosen Universitas Bina Sarana
Informatika, dosen Universitas Mercu Buana, mahasiswa Universitas Bina
Sarana Informatika, dan mahasiswa Universitas Mercu Buana. 26 Jurnal Pengabdian Masyarakat Formosa (JPMF)
Vol. 2 No 1, 2023: 21 - 28 Dapat dituliskan secara spesifik hasil dari pengabdian ini yang akan
dilakukan bersama dengan dunia industri adalah sebagai berikut: 1. Menciptakan peluang usaha yang memiliki prospek di tengah pandemi
bagi lulusan baru dengan sistem kemitraan bersama dunia industri. 1. Menciptakan peluang usaha yang memiliki prospek di tengah pandemi
bagi lulusan baru dengan sistem kemitraan bersama dunia industri. g
g
2. Tetap terbukanya lapangan pekerjaan bagi fresh graduate karena dunia
usaha masih tetap beroperasi di masa pandemi. 2. Tetap terbukanya lapangan pekerjaan bagi fresh graduate karena dunia
usaha masih tetap beroperasi di masa pandemi. 3. Terwujudnya tempat belajar secara langsung di masyarakat bagi
mahasiswa untuk mempelajari tentang strategi pemecahan kasus
terhadap perekonomian yang tidak pasti di tengah pandemi. 4. Pengabdian kepada masyarakat bagi para dosen dalam rangka
memperkenalkan sistem belanja baru di masa pandemi bagi para
konsumen dengan memberikan edukasi tentang cara belanja yang aman. 5. Kerjasama antar kampus bisa terjadi karena adanya konsorsium
penelitian sebagai bentuk tridharma di kampus lain. 6. HASIL DAN PEMBAHASAN Transfer teknologi dari perguruan tinggi kepada dunia industri dalam
bentuk purwarupa yang efektif dan efisien agar tetap bertahan
melakukan aktivitas usaha di tengah pandemi. 7. Terciptanya program studi baru untuk membantu mendukung
melakukan studi terhadap perkembangan inovasi seputar penanganan
dampak pandemi yang diakui oleh pihak terkait. 7. Terciptanya program studi baru untuk membantu mendukung
melakukan studi terhadap perkembangan inovasi seputar penanganan
dampak pandemi yang diakui oleh pihak terkait. KESIMPULAN DAN REKOMENDASI Dari hasil diskusi dapat diketahui bahwa masyarakat antusias, terutama
yang bermukim di perumahan karena dengan sistem online to offline (sistem
pesan online, barang datang offline, konsumen bisa memilih). Untuk kendaraan
dapat mengirit bahan bakar karena menggunakan mesin tenaga surya. Apabila
program ini bisa berlanjut maka dapat dipastikan Indonesia dapat mengurangi
polusi udara dari asap kendaraan. Dimana Indonesia dengan iklim nya yang
panas dapat menghasilkan energi surya secara terus-menerus. Oleh karena itu
program ini di rekomendasikan untuk di laksanakan ke depannya. Beberapa
program yang mungkin bisa diterapkan. UCAPAN TERIMA KASIH Pertama-tama saya sangat berterima kasih kepada rekan saya, bpk
Anantha Zakharia yang dari awal pengabdian telah menyusun proposal ini. Bpk Irvan Hermala atas ide terbarunya untuk menggunakan kendaraan tenaga
surya, serta adik-adik mahasiswa untuk pelaksanaan di lapangan. Dimana
dalam pelaksanaannya adik-adik mahasiswa ini ikut serta dalam kegiatan
kampus merdeka dalam hal penelitian energi tenaga surya. Jurnal ini tidak mungkin dapat saya selesaikan sendiri tanpa adanya
bantuan dari rekan-rekan yang saya sebutkan di atas. Walaupun dalam
pelaksanaannya tidak sesuai yang di targetkan karena besarnya dana yang di
perlukan, ke depan semoga bisa dijadikan acuan untuk pengabdian berikutnya. 27 Zakharia, Hermalam, Maharani DAFTAR PUSTAKA Arikunto, Suharsimi. (2002) Prosedur Penelitian Suatu Pendekatan Praktek
sebagai acuan pengambilan jumlah sampel minimum. Rineka Cipta Badan Pusat Statistik. (2021). Angkatan Kerja. https://www.bps.go.id Hermala, Irvan. (2020). Pembangkit Listrik Tenaga Surya. Jakarta. Univ.Mercu
Buana Program Pasca Sarjana Kemenristekdikti. (2019). Data
Mahasiswa
jurusan
Ekonomi. https://dikti.kemdikbud.go.id\ Zakharia, Anantha. (2012). Analisis Kualitas Produk dan Kualitas Pelayanan
dan Kepuasan Pelanggan, Pengaruhnya terhadap Loyalitas Pelanggan
KIKI Catering. Jakarta. Univ.KRISNADWIPAYANA Pogram Pasca
Sarjana 28
|
https://openalex.org/W4385071094
|
https://www.researchsquare.com/article/rs-2338043/latest.pdf
|
English
| null |
Rigid–flexible–thermal coupling dynamics of a hub and multiplate system considering frictional contact
|
Multibody system dynamics
| 2,023
|
cc-by
| 19,581
|
Rigid-§exible-thermal coupling dynamics of hub and
multi-plate system considering frictional contact Tingting Yuan
Shanghai Jiao Tong University
Bo Lei
Shanghai Jiao Tong University
Jinyang Liu ( liujy@sjtu.edu.cn )
Shanghai Jiao Tong University
Yunli Wu
Beijing Institute of Control Engineerin Abstract A geometric nonlinear modeling approach for strong rigid-flexible-thermal coupling dynamics of the hub and
multi-plate system considering frictional contact is proposed. Based on the absolute nodal coordinate formulation
(ANCF), a thermal integrated ANCF thin plate element is developed, where the temperature field is expressed with
Taylor polynomials to yield two-dimensional heat conduction equations. Different from the traditional coupling
formulations, the influences of the attitude motion and structural deformation on the intensity of the solar radiation,
the geometric nonlinearity of the plate as well as the frictional contact are taken into account. The normal contact is
formulated by the penalty method, and the tangential friction considers the stick–slip transition. To solve the strong
rigid-flexible-thermal coupling equations, a novel numerical method combining the modified central difference
approach and the generalized- method is proposed. Two validations are performed to verify the proposed
thermal-structural coupling model, which proves the importance of the geometric nonlinearity and can capture the
thermally induced vibration. Then the thermal-dynamic coupling analysis for the satellite and solar array multibody
system in thermal environment is carried out. The dynamic characteristics of the thermally induced vibration can be
successfully revealed by the rigid-flexible-thermal coupling model. Furthermore, it is indicated that the influence of
contact and thermal load on the nonlinear behavior of the solar array deployment is essential, which demonstrates
the feasibility of the proposed approach. Keywords Geometric nonlinear formulation · Rigid-flexible-thermal coupling dynamics · Thermally induced
vibration · Frictional contact Rigid-flexible-thermal coupling dynamics of hub and multi-plate system
considering frictional contact Tingting Yuan1 · Bo Lei1 · Jinyang Liu1 · Yunli Wu2 Tingting Yuan1 · Bo Lei1 · Jinyang Liu1 · Yunli Wu2 Research Article License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Additional Declarations: No competing interests reported. Version of Record: A version of this preprint was published at Multibody System Dynamics on July 21st,
2023. See the published version at https://doi.org/10.1007/s11044-023-09925-5. ,
g
g,
g
g
y,
g
,
2
Beijing Institute of Control Engineering & Key Laboratory of Space Intelligent Control Technology, Beijing 1 jy
j
1
Department of Engineering Mechanics, Key Laboratory of Hydrodynamics (Ministry of Education), School of
Architecture Ocean and Civil Engineering Shanghai Jiao Tong University Shanghai 200240 China j
artment of Engineering Mechanics, Key Laboratory of Hydrodynamics (Ministry of Education), School of Naval
hi
O
d Ci il E
i
i
Sh
h i Ji
T
U i
i
Sh
h i 200240 Chi 1
Department of Engineering Mechanics, Key Laboratory of Hydrodynamics (Ministry of Education liujy@sjtu.edu.cn
1
Department of Engineering Mechanics, Key Laboratory of Hydrodynamics (Ministry of Education), School of Naval J. Liu
liujy@sjtu.edu.cn J. Liu
liujy@sjtu.edu.cn
1
Department of Engineering Mechanics, Key Laboratory of Hydrodynamics (Ministry of Education), School of Naval
Architecture, Ocean and Civil Engineering, Shanghai Jiao Tong University, Shanghai 200240, China
2
Beijing Institute of Control Engineering & Key Laboratory of Space Intelligent Control Technology, Beijing 100190, China 1 Introduction The Absolute Nodal Coordinate Formulation (ANCF) proposed by Shabana [14] has been widely used for
simulation of plate with large deformation. Since the transverse shear deformable of thin plate can be neglected,
based on Kirchhoff assumptions, Dmitrochenko et al. [15,16], Dufva [17] and Ren [18] parameterized the plate
elements using slopes in the element mid-surface direction only. Schwab et al. [19] and Sereshk and Mahmoud [20]
compared the thin plate elements against the plate element based on the conventional finite element approach. Hyldahl et al. [21] also compared the convergence of rectangular thin plate elements in different mesh
configurations and load conditions. In ANCF, the gradient vectors are used to describe the nodal rotations instead
of the rotation parameters, avoiding the singularity problem and the interpolation problem of the finite rotation
variables [22-24]. Hence, ANCF is suitable for the geometric nonlinear dynamics modeling of rigid-flexible
coupled or rigid-flexible-thermal coupled multibody systems. Based on ANCF shear deformable plate element,
Shen et al. [25] proposed a coupled thermal-structural model of a laminated composite plate. Cui and Yu proposed
a novel method of the thermo-mechanical coupled analysis based on the unified description [26], and further
developed a thermal integrated ANCF thin plate, which depicts the displacement and the temperature field
integratedly [27]. Since the contact may occur during the deployment of a hub and multi-plate system, it is necessary to consider
the effect of frictional contact in the rigid-flexible-thermal coupling modeling. The key to modeling the contact
dynamics of multibody systems lies in the treatment of contact interface nonlinearities, which involves contact
detection, contact discretization, enforcement of contact conditions and friction force model [28-31]. In recent
years, some researchers have already developed several different formulations for modeling the frictional contact
problem in multibody systems. Konyukhov and Schweizerhof [32,33] developed a geometrically exact theory for
various 3D solid contact elements, including the normal, the tangential and the rotational interactions. Yu et al. [34]
established the frictional contact formulation between flexible and rigid bodies through the Hertz contact model
and the velocity-based friction model. Shi et al. [35] presented a rotation-free shell formulation and an extended
contact discretization for the dynamics of multibody systems with large deformations and frictionless contacts. Sun
et al. [36,37] developed a new 2D segment-to-segment algorithm of contact dynamics based on ANCF and mortar
method, including both frictionless and friction cases. Gay Neto et al. 1 Introduction Thermally induced vibrations of flexible spacecraft appendages may be initiated through the rapid changes of
thermal loading during the orbital eclipse transitions. The thermal gradient in the thickness direction is caused by
the difference of the heat flux applied on the upper and lower surfaces of the solar arrays, which may induce the
thermal deformation, thermal vibration and even influence the attitude motion of the satellite. Additionally, the
thermal-dynamic modeling of the spacecraft in many cases involves contact interaction between flexible
appendages undergoing large deformations and large overall motions. Therefore, an accurate rigid-flexible-thermal
coupling model is essential to guide the practical engineering applications, and this study remains a worthy and
challenging area. Concerning the investigation of the thermal-structural coupling effect, Thornton and Kim [1] developed an
analytical approach for determining the dynamic response of a flexible rolled-up solar array due to a sudden
increase in external heating, in which the rotational motion of the spacecraft was not taken into account. Johnston
and Thornton [2] presented an analytical model to investigate the effects of thermally induced vibrations of the
flexible appendages on the rotational motion of the spacecraft, and then further carried out the experiment
investigation on the structural-thermal coupling dynamics [3]. Fan and Liu [4] developed a 2D
thermal-rigid-flexible coupling model to investigate the thermally induced vibration of hub-beam system. Shen et al. analyzed the structural-thermal coupling performance for a space thin-walled beam [5] and a spinning spacecraft
with axial boom [6], and further conducted a stability analysis for the thermally induced fluttering phenomena [7]. Čepon et al. [8] proposed a new model for a coupled thermal-structural analysis of the bimetallic strip based on the
ANCF. Liu et al. [9-11] carried out a thermal-structural analysis for a flexible spacecraft with a single or double
solar panels, in which the coupling effect among attitude motion, structural deformation and thermal loading are
taken into account. Li et al. [12] investigated rigid-flexible-thermal coupling dynamics for a hub-beam with
clearance joint considering torsional spring, latch mechanism, and attitude controller. Since the solar arrays are thin plates, it is necessary to extend the thermal–structural coupling investigation to
plate structures. Liu et al. [13] investigated dynamic performance of a hub-plate system. The influence of the
rotational motion on the intensity of the heat flux was included in the dynamic model. However, the geometric
nonlinearity as well as the contact between the flexible bodies were not considered. 1 Introduction [38] proposed a formulation to handle
pointwise frictional contact interaction of beam-shell using the degeneration of the local contact problem. Tang and
Liu [39] modeled the frictional contact in sliding joints with clearances, and generalized the regularized Coulomb
friction law into ANCF beam and rigid hole considering the stick–slip transition. Lei et al. [40] presented a strong
coupling modelling method for the rigid bodies and the large deformed beams in contact interaction with the
granular matter to simulate the flexible brush sampling process. To avoid mutual penetration of plate element with
large deformation, a mixed method combining the node-to-surface, edge-to-surface and surface-to-surface contact
elements based on ANCF was proposed by the authors [41,42]. Nevertheless, few studies have considered the
frictional contact in the rigid-flexible-thermal coupling modeling. In this paper, a strong rigid-flexible-thermal coupling dynamic model for hub and multi-plate system is
proposed considering the geometric nonlinearity and frictional contact. The remaining part of the paper is
organized as follows. In Section 2, the finite element discretized dynamic variational equations of the rectangular
plate with thermoelasticity are presented based on ANCF. Then, the contact interactions between rigid bodies and
plates are addressed by the surface-to-surface and surface-to-rigid surface frictional contact elements. By leading
into the constraint equations of the multibody system, the rigid-flexible coupling dynamic equations are derived. In
Section 3, the rigid-flexible-thermal coupling equations are derived, which are composed of the heat conduction
equations and the Lagrange dynamic equations of the first kind. A new numerical method combining modified
central difference method and generalized- method is proposed in Section 4, which can be used to solve the heat
conduction equations and the dynamic equations simultaneously. Two validations are carried out to validate the
correctness of the proposed thermal-structural coupling model in Section 5. In order to consider the influence of
attitude motion and structural deformation on the intensity of the solar radiation, the relation between the heat flux
and the absolute coordinates is derived in Section 6. Then the thermal-dynamic coupling analysis for the satellite
and solar array multibody system in thermal environment is conducted. Summary and conclusions are presented in
Section 7. 2 Equations of motion of rigid body and plate system In this section, the absolute nodal coordinate formulation of a thin plate element with thermoelasticity is introduced,
and then equations of motion of hub and multi-plate system with frictional contact are derived. 2.1 Variational equations of motion of the plate based on the thermal integrated ANCF thin plate Based on Kirchhoff assumption the virtual work done by the elastic force of the element i can be written as j
j,
j
y
j,
j
y
j,
chhoff assumption, the virtual work done by the elastic force of the element i can be written as Based on Kirchhoff assumption, the virtual work done by the elastic force of n, the virtual work done by the elastic force of the element i can be written as
e
e
a
b
h
h
T
ei
x
y
z
T
U
0
0
2
/
2
/
T
d
d
d
α
ε
D
ε
,
(3)
e
e
a
b
h
h
T
ei
x
y
z
T
U
0
0
2
/
2
/
T
d
d
d
α
ε
D
ε
,
(3) (3) where D represents the matrix of modulus, and
T]
0
[
y
x
T
α
represents the matrix of the thermal
expansion coefficient, and represents the Green-Lagrange strain vector of an arbitrary point of the plate, which is
given by where D represents the matrix of modulus, and
T]
0
[
y
x
T
α
represents the matrix of the thermal
expansion coefficient, and represents the Green-Lagrange strain vector of an arbitrary point of the plate, which is
given by κ
ε
ε
z
0
,
(4) (4) where and represent the in-plane Green-Lagrange strain vector and the curvature vector, respectively, which
can be given as
n
r
n
r
n
r
κ
r
r
r
r
r
r
ε
T
T
T
T
T
2
1
T
2
1
0
2
1
,
1
1
xy
yy
xx
y
x
y
y
x
x
n
,
(5) (5) where n is a vector normal to the mid-surface of the plate. T
is the temperature variation, which can be
expressed with Taylor polynomials as where n is a vector normal to the mid-surface of the plate. 2.1 Variational equations of motion of the plate based on the thermal integrated ANCF thin plate T
is the temperature variation, which can be
expressed with Taylor polynomials as
t
y
x
T
z
t
y
x
zT
t
y
x
T
t
z
y
x
T
m
m
,
,
,
,
,
,
)
,
,
,
(
1
0
,
(6)
. ,
,1
,
,
,
,
,
)
,0,
,
(
,
,
0
0
m
k
z
t
z
y
x
T
t
y
x
T
t
y
x
T
t
y
x
T
z
k
k
k
，
(7) (6) (7) Following Eq. (6), the original three-dimensional temperature field can be reduced to a two-dimensional one. Considering that the thickness of the thin plate is small and the higher order terms of Eq. (6) can be neglected, thus
T
is reduced as
t
y
x
zT
t
y
x
T
t
z
y
x
T
,
,
,
,
)
,
,
,
(
1
0
,
(8) (8) Substituting Eqs. (4) and (6) into Eq. (3), the virtual work done by the elastic force can be rewritten as
d
d
0
0
T
3
12
1
0
T
0
e
e
a
b
ei
x
y
h
h
U
Dκ
κ
Dε
ε
(9 s. (4) and (6) into Eq. (3), the virtual work done by the elastic force can be rewritten as
Substituting Eqs. (4) and (6) into Eq. 2.1 Variational equations of motion of the plate based on the thermal integrated ANCF thin plate 4,
,1
,
] T
T
,
T
,
T
,
j
xy
ij
y
ij
x
ij
r
r
r
, and
x
ij
x
ij
/
,
r
r
,
y
j
i
y
ij
/
,
r
r
,
y
x
ij
xy
ij
/
2
,
r
r
. where
4,
,1
,
]
[
T
T
,
T
,
T
,
T
j
xy
ij
y
ij
x
ij
ij
ij
r
r
r
r
q
, and
x
ij
x
ij
/
,
r
r
,
y
j
i
y
ij
/
,
r
r
,
y
x
ij
xy
ij
/
2
,
r
r
. Based on Kirchhoff assumption, the virtual work done by the elastic force of the element i can be written as where
4,
,1
,
]
[
T
T
,
T
,
T
,
T
j
xy
ij
y
ij
x
ij
ij
ij
r
r
r
r
q
, and
x
ij
x
ij
/
,
r
r
,
y
j
i
y
ij
/
,
r
r
,
y
x
ij
xy
ij
/
2
,
r
r
. 2.1 Variational equations of motion of the plate based on the thermal integrated ANCF thin plate (3), the virtual work done by the elastic force can be rewritten as
d
d
0
0
T
3
12
1
0
T
0
e
e
a
b
ei
x
y
h
h
U
Dκ
κ
Dε
ε
d
d
0
0
T
3
12
1
0
T
0
e
e
a
b
ei
x
y
h
h
U
Dκ
κ
Dε
ε
(9) (9) ,
d
d
d
d
0
0
1
T
3
12
1
0
0
0
T
0
e
e
e
e
a
b
T
a
b
T
x
y
T
h
x
y
T
h
α
D
κ
α
D
ε
(9) Variation of and lead to Variation of and lead to Variation of and lead to of and lead to i
i
q
G
κ
q
H
ε
,
0
,
(10) (10)
x
y
y
x
y
y
x
x
S
r
S
r
S
r
S
r
H
T
T
T
T
,
(11)
Y
n
κ
S
n
S
n
S
n
Y
r
Y
r
Y
r
T
2
T
T
T
T
T
1
2
1
2
1
n
n
n
G
xy
yy
xx
xy
T
yy
xx
,
(12)
y
xr
r
n
~
,
x
y
y
x
S
r
S
r
Y
~
~
,
(13)
x
y
y
x
y
y
x
x
S
r
S
r
S
r
S
r
H
T
T
T
T
,
(11) (11) Y
n
κ
S
n
S
n
S
n
Y
r
Y
r
Y
r
T
2
T
T
T
T
T
1
2
1
2
1
n
n
n
G
xy
yy
xx
xy
T
yy
xx
,
(12) (12) y
xr
r
n
~
,
x
y
y
x
S
r
S
r
Y
~
~
,
(13) (13) in which
xr~ and
yr~ are the 3×3 skew-symmetric matrices corresponding to rx and ry, respectively. 2.1 Variational equations of motion of the plate based on the thermal integrated ANCF thin plate A thin rectangular plate element with length ae, width be and height h, is shown in Fig. 1. As can be seen in the
figure, O-XYZ represents the inertial coordinate system, and Oe-XeYeZe represents the element coordinate system. )
(t
r
k
)
( 0t
r
0k
ea
eb
h
Reference Configuration
Current Configuration
O
X
Y
Z
Oe
Xe
Ye
Ze
Fig. 1 A thin rectangular plate element of ANCF )
(t
r
k
)
( 0t
r
0k
ea
eb
h
Reference Configuration
Current Configuration
O
X
Y
Z
Oe
Xe
Ye
Ze
Fig. 1 A thin rectangular plate element of ANCF )
(t
r
k
)
( 0t
r
0k
ea
eb
h
Reference Configuration
Current Configuration
O
X
Y
Z
Oe
Xe
Ye
Ze
Fig. 1 A thin rectangular plate element of ANCF Current Configuration Current Configuration X Fig. 1 A thin rectangular plate element of ANCF The global position vector of an arbitrary point of element i in the mid-surface can be defined using the linear
combination of the element shape function matrix S and the global nodal coordinate vector q, which is given by
q
B
q
q
S
r
i
i
i
y
x
,
)
,
(
,
(1) q
B
q
q
S
r
i
i
i
y
x
,
)
,
(
,
(1) (1) where qi represents the element nodal coordinate vector, and x and y are the spatial coordinates defined in the
element coordinate system, respectively. Bi is a Boolean matrix. In the absolute nodal coordinate formulation, qi
contains the global position vectors and slopes which are the spatial derivatives of the position vectors. The element
nodal coordinate vector can be written as
T
T
4
T
3
T
2
T
1
i
i
i
i
i
q
q
q
q
q
,
(2) (2) where
4,
,1
,
]
[
T
T
,
T
,
T
,
T
j
xy
ij
y
ij
x
ij
ij
ij
r
r
r
r
q
, and
x
ij
x
ij
/
,
r
r
,
y
j
i
y
ij
/
,
r
r
,
y
x
ij
xy
ij
/
2
,
r
r
. 2.2 Variational equations of motion of the rigid body attached to the plate Consider a rigid body with mass mc, the 3×3 symmetric inertia matrix
c
J, which is defined in its centroidal frame
Oc-XcYcZc, as well as the position vector of the centroid rc defined in the inertial frame, and the angular velocity
c
ω, which is defined in Oc-XcYcZc. The variational equations of motion take the form
0
~
T
T
c
c
c
c
c
c
c
c
c
c
c m
L
J
J
F
r
r
,
(17) (17) where
c
F and
c
L are the vectors of the external force and torque applied on the rigid body defined in the inertial
frame and its centroidal frame, respectively. Defining
T
T
T
]
[
c
c
c
θ
r
q
as the generalized coordinate vector of the rigid body, where c is the Cardan
orientation angle vector of Oc-XcYcZc with respect to the inertial frame, the relation among
c
ω,
c
ω,
c
θ,
c
θ
are given by [43,44] c
c
c
θ
K
ω
,
c
c
c
c
c
θ
K
θ
K
ω
,
(18) (18) 2.1 Variational equations of motion of the plate based on the thermal integrated ANCF thin plate Substituting Eq. (10) into Eq. (9), the elastic force of the element is given by
d
d
d
d
d
d
/
1
T
3
1
0
T
0
0
T
3
12
1
0
T
T
e
e
e
e
e
e
a
b
T
a
b
T
a
b
i
ei
ei
x
y
T
h
x
y
T
x
y
h
h
U
α
D
G
α
D
H
Dκ
G
Dε
H
q
Q
(14)
d
d
0
0
T
3
12
1
0
T
e
e
a
b
x
y
h
h
Dκ
G
Dε
H
(14) (14)
. d
d
d
d
d
d
0
0
1
T
3
12
1
0
0
0
T
0
0
12
1
0
e
e
e
e
e
e
a
b
T
a
b
T
x
y
T
h
x
y
T
x
y
h
h
α
D
G
α
D
H
Dκ
G
Dε
H
(14) . d
d
d
d
0
0
1
T
3
12
1
0
0
0
T
e
e
e
e
a
b
T
a
b
T
x
y
T
h
x
y
T
α
D
G
α
D
H . d
d
d
d
0
0
1
T
3
12
1
0
0
0
T
e
e
e
e
a
b
T
a
b
T
x
y
T
h
x
y
T
α
D
G
α
D
H Accordingly, the elastic force can be written as
T can be written as Accordingly, the elastic force can be written as Accordingly, the elastic force can be written as
i
ei
i
e
Q
B
Q
T
. where Kc is the function of c. where Kc is the function of c. Thus, Eq. (17) can be rewritten as Thus, Eq. (17) can be rewritten as Thus, Eq. (17) can be rewritten as
0
c
c
c
T
c
Q
q
M
q
,
(19)
0
c
c
c
T
c
Q
q
M
q
,
(19) (19) where
c
c
T
c
c
c
m
K
J
K
0
0
I
M
3
3
3
3
3
3
,
(20)
c
c
c
c
c
c
c
T
c
c
c
K
J
J
L
K
F
Q
~
. (21) (20) (21) .3 Frictional Contact formulations of rigid body and plate 2.1 Variational equations of motion of the plate based on the thermal integrated ANCF thin plate (15) (15) Defining as the mass density, and g as gravitational acceleration, and as the structural damping coefficient,
the variational equations of motion read
0
T
g
e
Q
Q
q
C
q
M
q
,
(16) (16) where
i
a
b
i
e
e
x
y
h
B
S
S
B
M
)
d
d
( 0
0
T
T
is the constant generalized mass matrix, and
M
C
is the damping
matrix, and
)
d
d
( 0
0
T
T
e
e
a
b
i
g
x
y
h
g
S
B
Q
is the generalized gravitational force vector. 2.3 Frictional Contact formulations of rigid body and plate In this study, two forms of contact are taken into account, i.e., plate-to-plate contact and plate-to-rigid body contact. The normal contact is formulated using the penalty method [41,45], and the tangential contact is modeled by the
Coulomb friction law with a penalty regularization considering the stick-slip transition [42]. The virtual work of the
contact force is
otherwise
0
if
0
,
d
T
N
T
N
cf
cf
g
W
f
f
u
,
(22) (22) where gN is the normal penetration, and is the contact area. ucf is the relative displacement vector of a contact
pair, and fN, fT are the normal and tangential force vectors of a contact pair, respectively. Contact between the
contact pair occurs only if
0
N
g
. where gN is the normal penetration, and is the contact area. ucf is the relative displacement vector of a contact
pair, and fN, fT are the normal and tangential force vectors of a contact pair, respectively. Contact between the
contact pair occurs only if
0
N
g
. 2.3.1 Contact between plates 2.3.1 Contact between plates S(Sa)
P
Pa
Slave
Master
x S
h S
h M
x M
n
Integration
points
O
X
Y
Z
Fig. 2 Geometry and kinematics of the STS contact element Integration
points Fig. 2 Geometry and kinematics of the STS contact element As shown in Fig. 2, the contact is checked between each integration points of the slave surface and the master
surface, which is named as STS contact element. For the quadrilateral elements, the nodal vector for the STS
contact element can be given by 1
48
1
48
T
T
T
,
,
s
ec
m
ec
s
ec
m
ec
ec
q
q
q
q
q
. (23) (23) The normal penetration can be defined as The normal penetration can be defined as
2
T
S
M
P
S
N
h
h
g
n
r
r
,
(24) (24) where hM and hS are the thicknesses of the master and slave surfaces, respectively. n is the unit normal vector of the
STS contact element, which can be obtained by Eq. (13). 2.3 Frictional Contact formulations of rigid body and plate )
,
(
)
,
(
m
eca
M
Pa
M
Pa
ma
m
ec
M
P
M
P
m
P
q
S
q
S
r
h
x
h
x
(29) Accordingly, the incremental relative displacement vector on the current tangent plane is al relative displacement vector on the current tangent plane is
cf
T
u
nn
I
g
T
3
.
cf
T
u
nn
I
g
T
3
. (30) (30) The return-mapping scheme [32] for the Coulomb friction law with a penalty regularization is applied to
measure the friction force, it introduces the trial tangential force vector as
T
T
Ta
tr
T
g
f
nn
I
f
T
3
,
(31) (31) where fTa is the real tangential force vector in the previous step, and T is the tangential penalty parameter. According to the yield function of Coulomb friction law, the real tangential force of the contact pair SP can be
determined as
sliding
,0
if
,
sticking
,0
if
,
N
s
tr
T
tr
T
tr
T
N
d
N
s
tr
T
tr
T
T
f
f
f
f
f
f
f
f
f
,
(32) (32) where s and d are the static and dynamic frictional coefficient. where s and d are the static and dynamic frictional coefficient. Then, the element residual vector of the STS contact element can be summarized as
d
T
T
N
STS
ec
f
f
S
Q
. (33) (33) 2.3.2 Contact between plate and rigid body 2.3.2 Contact between plate and rigid body C
Xc
Yc
Zc
S(Sa)
Slave
x S
h S
Integration
points
Master
O
X
Y
Z
n
P
Pa
n'
Fig. 3 Geometry and kinematics of STRS contact element Integration
points Fig. 3 Geometry and kinematics of STRS contact element As shown in Fig. 3, the contact is checked between each integration points of the slave plate surface and the master
rigid surface, which is named as STRS contact element. 2.3 Frictional Contact formulations of rigid body and plate Thus, the variation of the relative displacement vector of
the contact pair SP is written as where hM and hS are the thicknesses of the master and slave surfaces, respectively. n is the unit normal vector of the
STS contact element, which can be obtained by Eq. (13). Thus, the variation of the relative displacement vector of
the contact pair SP is written as ec
cf
P
S
cf
q
S
r
r
u
,
(25) (25) here Scf is the shape function matrix of the contact element, which can be defined as
)
,
(
)
,
(
S
S
s
M
M
m
cf
h
x
h
x
S
S
S
,
(26) (26) where Sm and Ss are the shape functions of the master and slave surfaces, respectively. x and h are the convective
coordinates defined in the local surface element coordinate system. Based on the penalty method, the normal contact force of the contact pair SP is formulated as
n
f
N
N
N
g
,
(27) Based on the penalty method, the normal contact force of the contact pair SP is formulated as
n
f
N
N
N
g
,
(27) (27) In order to describe the stick-slip transition, it requires the history variables from the previous step to describe
the incremental nodal displacement [39]. Here, the points related to the previous step are represented by a subscript
(·)a. As shown in Fig. 2, the contact point of the slave node Sa on the master surface is Pa in the previous step. Then,
the incremental relative displacement vector of the contact pair SP is P
S
cf
r
r
u
,
(28) P
S
cf
r
r
u
,
(28) (28) P
S
cf
r
r
u
,
(28) where where (29) . )
,
(
)
,
(
m
eca
M
Pa
M
Pa
ma
m
ec
M
P
M
P
m
P
S
q
S
q
S
r
h
x
h
x
(29) . 2.4 Rigid-flexible coupling equations of the rigid body and plate system The kinematics constraints between the rigid body and the plate include nj fixed joints and nd driving constraints. The constraint equations are given by
0
q
q
Φ
)
,
,
(
t
c
, the mixed differential-algebraic equations [46] can be given
by 0
Φ
Q
λ
Φ
q
C
q
M
q
d
d
d
d
d
d
T
,
(39) (39) where the system generalized coordinate vector is given by
T
T
T
]
[
q
q
q
c
d
, and the generalized mass, damping
and force matrices take the form where the system generalized coordinate vector is given by
T
T
T
]
[
q
q
q
c
d
, and the generalized mass, damping
and force matrices take the form
M
0
0
M
M
6
6
q
q
n
n
c
d
,
(40)
C
0
0
0
C
6
6
6
6
q
q
n
n
d
,
(41)
cf
e
g
c
d
Q
Q
Q
Q
Q
,
(42)
M
0
0
M
M
6
6
q
q
n
n
c
d
,
(40)
C
0
0
0
C
6
6
6
6
q
q
n
n
d
,
(41)
cf
e
g
c
d
Q
Q
Q
Q
Q
,
(42) (40) (41) cf
e
g
c
d
Q
Q
Q
Q
Q
,
(42) (42) and nq is the number of the generalized coordinates of the plate, and is the vector of Lagrange multiplier
associated with the kinematics and driving constraints. and nq is the number of the generalized coordinates of the plate, and is the vector of Lagrange multiplier
associated with the kinematics and driving constraints. 2.3 Frictional Contact formulations of rigid body and plate Thus, the nodal vector for the STRS contact element can
be given by 1
48
1
6
T
T
T
,
,
s
ec
c
s
ec
c
ec
q
q
q
q
q
. (34) (34) The normal penetration can be obtained by
2
T
M
P
S
N
h
g
n
r
r
,
(35) (35) where
n
A
n
c
is the unit normal vector of the STRS contact element, and Ac is the direction cosine of the rigid
body, and n is the unit normal vector of the rigid surface, which is defined in the body-fixed frame Oc-XcYcZc. Then, the shape function matrix of the STRS contact element is where
n
A
n
c
is the unit normal vector of the STRS contact element, and Ac is the direction cosine of the rigid
body, and n is the unit normal vector of the rigid surface, which is defined in the body-fixed frame Oc-XcYcZc. Then, the shape function matrix of the STRS contact element is
s
M
M
m
cf
S
S
S
)
,
(
h
x
,
(36) (36) where
]
~
[ 3
c
P
c
s
K
ρ
A
I
S
is the shape functions of the slave rigid surface, and
P
ρ is the position vector of the
point P, which is defined in Oc-XcYcZc. Similar to the STS contact element, the element residual vector of the
STRS contact element can be obtained as where
]
~
[ 3
c
P
c
s
K
ρ
A
I
S
is the shape functions of the slave rigid surface, and
P
ρ is the position vector of the
point P, which is defined in Oc-XcYcZc. Similar to the STS contact element, the element residual vector of the
STRS contact element can be obtained as
d
T
T
N
STRS
ec
f
f
S
Q
. (37) (37) Therefore, the generalized contact force vector relate to Eq. 2.3 Frictional Contact formulations of rigid body and plate (22) can be assembled as
2
1
T
T
c
c
n
STRS
ec
STRS
e
n
STS
ec
STS
e
cf
Q
B
Q
B
Q
,
(38) (38) where
STS
e
B
and
STRS
e
B
are the connectivity matrices of the STS and STRS contact elements, respectively. nc1
and nc2 are the numbers corresponding to STS and STRS contact elements, respectively. where
STS
e
B
and
STRS
e
B
are the connectivity matrices of the STS and STRS contact elements, respectively. nc1
and nc2 are the numbers corresponding to STS and STRS contact elements, respectively. where Tr is the reference temperature. The variational heat conductive equations can be written as The variational heat conductive equations can be written as
0
T
T
T
T
F
p
K
p
M
p
,
(45) (45) where ,
,
1
0
1
0
1
0
1
0
F
F
F
K
0
0
K
K
M
0
0
M
M
p
p
p
T
T
T
，
，
(46) (46)
0,1
,
d
d
T
0
0
k
x
y
c
b
e
e
a
b
k
k
N
N
M
,
(47) (47)
0,1
,
d
d
d
d
T
0
0
T
T
0
0
k
x
y
k
kh
x
y
k
b
e
e
e
e
a
b
y
y
x
x
a
b
k
k
N
N
N
N
N
N
K
,
(48) (48)
0,1
,
d
d
5.0
d
d
5.0
d
d
T
0
0
T
0
0
T
0
0
k
x
y
q
h
x
y
q
h
x
y
Q
b
e
e
e
e
e
e
a
b
k
a
b
k
a
b
T
k
k
N
N
N
F
,
(49)
0,1
,
d
d
5.0
d
d
5.0
d
d
T
0
0
T
0
0
T
0
0
k
x
y
q
h
x
y
q
h
x
y
Q
b
e
e
e
e
e
e
a
b
k
a
b
k
a
b
T
k
k
N
N
N
F
,
(49)
0,1
,
d
2
/
2
/
k
z
z
b
h
h
k
k
. (50) (50) In case that the temperature variable vector p is independent, the heat conductive equations are given by 0
F
p
K
p
M
T
T
T
. (51) (51) 3 Variational heat conductive equations As can be seen in the Fig. 4, the thermal load applied on the thin plate includes the heat flux acting on the upper
and lower surfaces. The variational heat conductive equations are given by
,
0
d
d
d
S
S
V
T
z
z
y
y
x
x
S
Tq
S
Tq
V
Q
T
T
k
T
T
k
T
T
k
T
T
c
(43) (43) ,
0
d
d
S
S
S
Tq
S
Tq
where c is the specific heat, and k is the conductivity coefficient, and QT is the intensity of heat source, and
q ,
q represent the heat flux of the upper and lower surfaces, respectively. where c is the specific heat, and k is the conductivity coefficient, and QT is the intensity of heat source, and
q ,
q represent the heat flux of the upper and lower surfaces, respectively. n
O
X
Y
Z
q
q
T
Q
Upper surface S +
Lower surface S −
Xe
Ye
Oe
Fig. 4 A thin plate applied with thermal load Upper surface S + Lower surface S − Fig. 4 A thin plate applied with thermal load The temperature variables are discretized as
1
,0
k
T
ik
k
Np
, where
1
,0
k
ik
p
represent the vectors of
temperature variables of element i. Defining
1
,0
k
k
p
as the global vectors of temperature variables, the
relation between the element vector and the global vector is given by
1
,0
k
k
ik
ik
p
B
p
, the absolute
temperature can be expressed as 1
0
zT
T
T
T
T
T
r
r
. (44) 1
0
zT
T
T
T
T
T
r
r
. (44) where Tr is the reference temperature. 4.1 Prediction of the initial value and the numerical parameters
]1
,0
[
,
. and the numerical parameters
]1
,0
[
,
. 4 Solution of the rigid-flexible-thermal coupling equations Combining Eqs. (39) and (51), the mixed algebraic equations are given by Combining Eqs. (39) and (51), the mixed algebraic equations are given by 0
F
p
K
p
M
Φ
Q
λ
Φ
q
C
q
M
Ψ
q
T
T
T
d
d
d
d
d
d
T
. (52) (52) A new numerical method combining the modified central difference approach and the generalized- method is
proposed to solve the strong rigid-flexible-thermal coupling equations. The heat conduction equations are solved by
modified central difference method, and the dynamic equations are solved by generalized- method [47], and
finally, Newton-Raphson algorithm is used to solve the combined nonlinear algebraic equations in Eq. (52). The
solution procedure is introduced as follows. 4.2 Transformation into nonlinear algebraic equations The dynamic equations in Eq. (39) can be transformed into nonlinear algebraic equations using generalized-
method
0
q
Φ
p
q
Q
λ
q
Φ
q
C
q
M
q
)
,
(
)
,
(
T
t
t
t
t
d
t
t
t
t
d
d
t
t
t
t
d
t
t
d
d
t
t
d
d
d
,
(60)
0
q
Φ
p
q
Q
λ
q
Φ
q
C
q
M
q
)
,
(
)
,
(
T
t
t
t
t
d
t
t
t
t
d
d
t
t
t
t
d
t
t
d
d
t
t
d
d
d
,
(60)
where
f
m
t
t
d
t
t
d
t 1
1
2
q
q
,
(61)
t
t
t
d
t
t
d
q
q
,
(62) (60) where
f
m
t
t
d
t
t
d
t 1
1
2
q
q
,
(61) (61)
t
t
t
d
t
t
d
q
q
,
(62) (62) The heat conduction equations in Eq. (51) are transformed into nonlinear algebraic equations by modified
central difference method The heat conduction equations in Eq. 4.2 Transformation into nonlinear algebraic equations (51) are transformed into nonlinear algebraic equations by modified
central difference method 0
p
q
Q
p
K
)
,
(
t
t
t
t
d
t
t
,
(63) (63) where T
T
t
K
M
K
,
(64) (64)
t
t
T
t
T
t
T
T
t
F
F
p
K
M
Q
1
1
. (65)
t
t
T
t
T
t
T
T
t
F
F
p
K
M
Q
1
1
. (65) (65) 4.3 Solution of the combined nonlinear algebraic equations 4.3 Solution of the combined nonlinear algebraic equations 4.3 Solution of the combined nonlinear algebraic equations 4.3 Solution of the combined nonlinear algebraic equations 4.1 Prediction of the initial value Assuming that
0
q
t
t
d
, we combine Taylor’s formula, generalized- method and modified central difference
method to predict
t
t
d
a
,
t
t
d
q
,
t
t
d
q
,
t
t
λ
,
t
t
p
, as follows: t
d
m
m
t
d
m
f
t
t
d
a
q
a
1
1
,
(53)
t
t
d
t
d
t
d
t
d
t
t
d
t
t
t
a
a
q
q
q
2
2
1
2
,
(54) t
d
m
m
t
d
m
f
t
t
d
a
q
a
1
1
,
(53)
t
t
d
t
d
t
d
t
d
t
t
d
t
t
t
a
a
q
q
q
2
2
1
2
,
(54)
t
t
d
t
d
t
d
t
t
d
t
t
a
a
q
q
1
,
(55) t
d
m
m
t
d
m
f
t
t
d
a
q
a
1
1
,
(53) (53)
t
t
d
t
d
t
d
t
d
t
t
d
t
t
t
a
a
q
q
q
2
2
1
2
,
(54) (54)
t
t
d
t
d
t
d
t
t
d
t
t
a
a
q
q
1
,
t
t
d
t
d
t
d
t
t
d
t
t
a
a
q
q
1
,
(55) (55) t
t
t
λ
λ
,
(56)
t
t
t
t
t
p
p
1
,
(57)
1
2 t
t
t
λ
λ
,
(56)
t
t
t
t
t
p
p
p
1
,
(57) t
t
t
λ
λ
,
(56) t
t
t
λ
λ
,
t
t
t
t
t
p
p
p
1
,
(57)
where
1
1
2
m
,
1
f
,
(58)
m
f
2
1
,
2
2
1
4
1
,
(59)
t
t
t
t
t
p
p
p
1
,
(57) where 1
1
2
m
,
1
f
,
(58) (58) m
f
2
1
,
2
2
1
4
1
,
(59) (59) and the numerical parameters
]1
,0
[
,
. 4.3 Solution of the combined nonlinear algebraic equations (70) (69)
0
0
q
Φ
λ
Ψ
q
t
t
k
d
t
t
d
)
(
T
. (70) (70) An improved estimate can be obtained as An improved estimate can be obtained as ,... 1
,0
,
)
(
)
(
)1
(
k
t
t
k
t
t
k
t
t
k
q
q
q
,
(71) (71) until the precision condition
t
t
k)
(
q
is satisfied, in which is the precision error. on condition
t
t
k)
(
q
is satisfied, in which is the precision error. Fig. 5 shows the computational flowchart of the new numerical method proposed in this paper. Compared with
the traditional uncoupled method, this method improves the accuracy of real-time simulation by solving the heat
conduction equations and the dynamic equations simultaneously. Additionally, considering the geometric
nonlinearities and contact issues, a variable-step-size control scheme [48] and a preconditioning strategy are
employed to improve efficiency. 5 Validations In this section, in order to verify the correctness of the proposed thermal-structural coupling model, simulation
experiments of a plate clamped at two sides and a cantilevered plate applied with thermal loads are carried out,
while the results from the commercial finite element software ABAQUS are taken as benchmarks. 4.3 Solution of the combined nonlinear algebraic equations At a iteration step k, the following
equation is solved for a correction
t
t
k
)
(
q
: Ψ
λ
Ψ
q
Ψ
p
Ψ
λ
q
p
q
1
)
(
)
(
)
(
)
(
t
t
t
t
d
t
t
t
t
k
t
t
k
d
t
t
k
t
t
k
,
(67) (67) where where
t
t
t
t
k
t
t
k
d
t
t
t
t
k
t
t
k
d
d
t
t
p
p
q
Q
K
p
p
q
Q
p
Ψ
)
,
(
)
,
(
)
(
)
(
)
(
)
(
,
(68) where
t
t
t
t
k
t
t
k
d
t
t
t
t
k
t
t
k
d
d
t
t
p
p
q
Q
K
p
p
q
Q
p
Ψ
)
,
(
)
,
(
)
(
)
(
)
(
)
(
,
(68)
t
t
t
t
k
t
t
k
d
t
t
t
t
k
t
t
k
d
d
t
t
p
p
q
Q
K
p
p
q
Q
p
Ψ
)
,
(
)
,
(
)
(
)
(
)
(
)
(
,
(68) (68)
t
t
d
t
t
k
t
t
k
d
t
t
k
d
t
t
d
t
t
k
t
t
k
d
d
t
t
d
t
t
k
t
t
k
d
d
d
t
t
d
d
d
q
p
q
Q
q
Φ
q
p
q
Q
q
λ
q
Φ
C
M
q
Ψ
q
q
)
,
(
)
,
(
)
(
)
(
)
(
)
(
)
(
)
(
)
(
T
,
(69)
0
0
q
Φ
λ
Ψ
q
t
t
k
d
t
t
d
)
(
T
. 4.3 Solution of the combined nonlinear algebraic equations According to Eqs. (60) and (63), the combined nonlinear algebraic equations are rewritten as
0. p
q
Q
p
K
q
Φ
p
q
Q
λ
q
Φ
q
C
q
M
λ
p
q
Ψ
q
)
,
(
)
,
(
)
,
(
,
,
,
T
t
t
t
t
d
t
t
t
t
d
t
t
t
t
d
d
t
t
t
t
d
t
t
d
d
t
t
d
d
t
t
t
t
t
t
d
t
t
t
t
d
(66) (66) The Newton-Raphson algorithm is used to find a solution of Eq. (53). At a iteration step k, the following
equation is solved for a correction
t
t
k
)
(
q
: The Newton-Raphson algorithm is used to find a solution of Eq. (53). 5.1 Simulation of a plate clamped at two sides (60)
t
t
t
t
p
p
,
t
t
d
t
t
d
t
t
d
q
q
q
,
,
Heat conduction equations in Eq. (66)
Combined nonlinear algebraic equations in Eq.(63)
Calculate data in Parallel:
Tangential stiffness matrices and force vectors: elastic
forces, contact forces / Heat-related matrices
Calculate in Eq.(67)
t
t
k
t
t
k
d
t
t
k
)
(
)
(
)
(
,
,
λ
q
p
Update data:
Kinematic and temperature variable vectors
Post-process:
Read the output data to calculate additional data
Predict the initial value: t = t + △t,
Obtain , , , , , in Eq.(53)~(57)
t
t
d
a
t
t
d
q
t
t
d
q
t
t
λ
t
t
p
0
q
t
t
d
t
t
k)
(
q
Calculate temperature variable
Generalized-method
Modified central difference method
end
t
t
Yes
Yes
No
No
Time integration
Newton-Raphson algorithm
Step control:
Calculate the estimation error, next step size
Reject? Yes
Update △t
No
Fig. 5
Computational flowchart for the proposed numerical method Calculate data in Parallel:
Tangential stiffness matrices and force vectors: elastic
forces, contact forces / Heat-related matrices
Calculate in Eq.(67)
t
t
k
t
t
k
d
t
t
k
)
(
)
(
)
(
,
,
λ
q
p
Update data:
Kinematic and temperature variable vectors
Post-process:
Read the output data to calculate additional data
t
t
k)
(
q
end
t
t
Yes
Yes
No
No
Step control:
Calculate the estimation error, next step size
Reject? Yes
No Post-process: p
Read the output data to calculate additional data Fig. 5 Computational flowchart for the proposed numerical method X
Y
Z
O
l/2
b
0
qT
P
l/2
Fig. 6 A plate clamped at two sides in thermal environment Fig. 6 A plate clamped at two sides in thermal environment As shown in Fig. 5.1 Simulation of a plate clamped at two sides As shown in Fig. 6, two sides of a plate are fixed on the inertial reference frame O – XYZ, and a constant heat flux
qT is applied to the upper surface of the plate. The initial temperature of the whole plate is set as 0 K. The geometric
property and material data of the plate are: length l = 4 m, width b = 2 m, thickness h = 0.01 m, mass density = 36.8 kg/m3, elastic modulus E = 1.93×109 Pa, Poisson ratio = 0.3, thermal expansion coefficient = 2.3×10-5
(1/K), conductivity coefficient k =1.5 W/(m.K), and specific heat c = 921 J/(Kg·K). In this simulation, the heat
flux of the upper and lower surfaces are q+ = qT cos and q− = 0, respectively, where qT = 1067 W/m2 and Input data:
Body meshed, Load, Constraints, Initial conditions
Integration Pre-process:
Constant matrices (mass matrix, damping matrix)
Calculate kinematic variable
Dynamic equations in Eq. (60)
t
t
t
t
p
p
,
t
t
d
t
t
d
t
t
d
q
q
q
,
,
Heat conduction equations in Eq. (66)
Combined nonlinear algebraic equations in Eq.(63)
Calculate data in Parallel:
Tangential stiffness matrices and force vectors: elastic
forces, contact forces / Heat-related matrices
Calculate in Eq.(67)
t
t
k
t
t
k
d
t
t
k
)
(
)
(
)
(
,
,
λ
q
p
Update data:
Kinematic and temperature variable vectors
Post-process:
Read the output data to calculate additional data
Predict the initial value: t = t + △t,
Obtain , , , , , in Eq.(53)~(57)
t
t
d
a
t
t
d
q
t
t
d
q
t
t
λ
t
t
p
0
q
t
t
d
t
t
k)
(
q
Calculate temperature variable
Generalized-method
Modified central difference method
end
t
t
Yes
Yes
No
No
Time integration
Newton-Raphson algorithm
Step control:
Calculate the estimation error, next step size
Reject? Yes
Update △t
No Input data:
Body meshed, Load, Constraints, Initial conditions
Integration Pre-process:
Constant matrices (mass matrix, damping matrix)
Calculate kinematic variable
Dynamic equations in Eq. 5.1 Simulation of a plate clamped at two sides 7, the time histories of the mid-surface temperature of point P and the temperature gradient
T1 =
z
T
/
of point P are given, and the results are in good agreement with the comparative results provided by
ABAQUS, which shows that the temperature field could be calculated precisely by the proposed method. To further investigate the geometric nonlinear effects, the deflections of point P in z direction using linear and
nonlinear models are compared in Fig. 8. Since the two edges of the plate are fixed, the thermal load leads to
compressive thermal stress. It is found that for the nonlinear model, the thermal force causes the softening of the plate, which induces large transverse deformations. However, the results obtained by the linear model are quite
different. Due to the neglect of the nonlinear stiffness matrices, the softening effect is not shown and the transverse
deformation is small. Furthermore, the results of this paper coincide with the results achieved by ABAQUS, which
validates the effectiveness of the proposed thermal integrated ANCF thin plate element in terms of geometric
nonlinearity. dates t e e ect ve ess o t e p oposed t e
a
teg ated
NC
t
p ate e e e t
te
s o geo et c
linearity. 0
1
2
3
4
5
0
4
8
12
16
T [K]
Time [s]
This paper
Abaqus
0
1
2
3
4
5
0
80
160
240
320
400
dT/dz [K/m]
Time [s]
This paper
Abaqus
(a)
(b)
Fig. 7 Time histories of (a) mid-surface temperature of point P; (b) temperature gradient of point P in z direction
0
1
2
3
4
5
-0.05
-0.04
-0.03
-0.02
-0.01
0.00
w [m]
Time [s]
This paper
Abaqus (nonlinear)
Abaqus (linear)
2
3
4
-1.8
-1.6
-1.4
x10-3
Fig. 8 Time history of the deflection of point P in z direction;
Simulation of a cantilevered plate
X
Y
Z
O
l
b
0
qT
P 0
1
2
3
4
5
0
4
8
12
16
T [K]
Time [s]
This paper
Abaqus
0
1
2
3
4
5
0
80
160
240
320
400
dT/dz [K/m]
Time [s]
This paper
Abaqus
(a)
(b)
Fig. 5.1 Simulation of a plate clamped at two sides 10(b),
we can obtain that the results of the linear and nonlinear models coincide well, it can be concluded that for
cantilevered plate without tip constraint, the use of linear model is computationally accurate and efficient. As shown in Fig. 9, the dynamic simulation of a cantilevered plate applied with thermal loads is performed. The
geometric property and material data of the plate are the same as the numerical example 5.1. The time history of the
temperature gradient of point P in z direction is given in Fig. 10(a), which shows that the results are in accordance
with the obtained results by ABAQUS. From the results of deflections of point P in z direction shown in Fig. 10(b),
we can obtain that the results of the linear and nonlinear models coincide well, it can be concluded that for
cantilevered plate without tip constraint, the use of linear model is computationally accurate and efficient. w [m]
0
1
2
3
4
5
0
80
160
240
320
400
dT/dz [K/m]
Time [s]
This paper
Abaqus
(a) 0
1
2
3
4
5
-0.14
-0.12
-0.10
-0.08
-0.06
-0.04
-0.02
0.00
w [m]
Time [s]
This paper
Abaqus (nonlinear)
Abaqus (linear)
0
1
2
3
4
5
0
80
160
240
320
400
dT/dz [K/m]
Time [s]
This paper
Abaqus
(a)
(b)
Fig. 10 Time histories of (a) temperature gradient; (b) deflection of point P in z direction; 0
1
2
3
4
5
-0.14
-0.12
-0.10
-0.08
-0.06
-0.04
-0.02
0.00
[
]
Time [s]
This paper
Abaqus (nonlinear)
Abaqus (linear)
(b) Fig. 10 Time histories of (a) temperature gradient; (b) deflection of point P in z direction; The thermal-structural analysis for the cantilevered plate applied with thermal loads is further conducted
through a comparative study of the thermal-structural coupling model and the constant heat flux model, where qT =
2525 W/m2 and , which ensures that q+ = 1067 W/m2. The time histories of the temperature gradient and
the deflection of point P in z direction are shown in Fig. 11, respectively. As can be seen in the figures, due to the
neglect of the elastic deformation of the constant heat flux model, such model cannot reveal the thermally induced
vibration effect, therefore the vibration amplitude of the temperature gradient and the deflection is constant. 5.1 Simulation of a plate clamped at two sides 7 Time histories of (a) mid-surface temperature of point P; (b) temperature gradient of point P in z direction 0
1
2
3
4
5
0
80
160
240
320
400
dT/dz [K/m]
Time [s]
This paper
Abaqus
(b) 0
1
2
3
4
5
0
4
8
12
16
T [K]
Time [s]
This paper
Abaqus
dT/dz [K/m]
(a) [
(b) Fig. 7 Time histories of (a) mid-surface temperature of point P; (b) temperature gradient of point P in z direction 0
1
2
3
4
5
-0.05
-0.04
-0.03
-0.02
-0.01
0.00
w [m]
Time [s]
This paper
Abaqus (nonlinear)
Abaqus (linear)
2
3
4
-1.8
-1.6
-1.4
x10-3
Fig. 8 Time history of the deflection of point P in z direction;
5.2 Simulation of a cantilevered plate
X
Y
Z
O
l
b
0
qT
P
Fig. 9 A cantilevered plate in thermal environment 0
1
2
3
4
5
-0.05
-0.04
-0.03
-0.02
-0.01
0.00
w [m]
Time [s]
This paper
Abaqus (nonlinear)
Abaqus (linear)
2
3
4
-1.8
-1.6
-1.4
x10-3
Fig. 8 Time history of the deflection of point P in z direction;
5.2 Simulation of a cantilevered plate
0
qT 0
1
2
3
4
5
-0.05
-0.04
-0.03
-0.02
-0.01
0.00
w [m]
Time [s]
This paper
Abaqus (nonlinear)
Abaqus (linear)
2
3
4
-1.8
-1.6
-1.4
x10-3 Fig. 8 Time history of the deflection of point P in z direction; Fig. 8 Time history of the deflection of point P in z direction; 5.2 Simulation of a cantilevered plate 5.2 Simulation of a cantilevered plate
X
Y
Z
O
l
b
0
qT
P
Fig. 9 A cantilevered plate in thermal environment 5.2 Simulation of a cantilevered plate Fig. 9 A cantilevered plate in thermal environment As shown in Fig. 9, the dynamic simulation of a cantilevered plate applied with thermal loads is performed. The
geometric property and material data of the plate are the same as the numerical example 5.1. The time history of the
temperature gradient of point P in z direction is given in Fig. 10(a), which shows that the results are in accordance
with the obtained results by ABAQUS. From the results of deflections of point P in z direction shown in Fig. 5.1 Simulation of a plate clamped at two sides On the
contrary, since the heat flux varies with the deformation of the plate, the vibration amplitudes of the temperature
gradient and the deflection obtained by the thermal-structural coupling model keep growing. In summary, the
proposed model takes into account the coupling effect of the heat flux and the elastic deformation, so that it is able
to capture the phenomenon of thermally induced vibration, which is hardly achieved in the commercial finite
element software ABAQUS. w [m]
0
4
8
12
16
20
0
80
160
240
320
400
dT/dz [K/m]
Time [s]
Thermal-structural coupling
Constant heat flux
(a) 0
4
8
12
16
20
-0.20
-0.16
-0.12
-0.08
-0.04
0.00
w [m]
Time [s]
Thermal-structural coupling
Constant heat flux
0
4
8
12
16
20
0
80
160
240
320
400
dT/dz [K/m]
Time [s]
Thermal-structural coupling
Constant heat flux
(a)
(b)
Fig. 11 Time histories of (a) temperature gradient; (b) deflection of point P in z direction; 0
4
8
12
16
20
-0.20
-0.16
-0.12
-0.08
-0.04
0.00
w [m]
Time [s]
Thermal-structural coupling
Constant heat flux
0
(b) Fig. 11 Time histories of (a) temperature gradient; (b) deflection of point P in z direction; 6.1 Numerical simulation of a satellite and solar array system in thermal environment
0
n
s
P
Sunlight
C
Solar array
Satellite
n
O
X
Y
Z
Xc
Yc
Zc
Fig. 12 A satellite and solar array system Satellite Yc Sunlight Fig. 12 A satellite and solar array system A satellite and solar array system applied with solar radiation fluxes is shown in Fig. 12. Since the distribution of
solar radiation input is affected by various factors, including its position, posture and configuration, the absorption
of incident solar rays needs to further evaluated by heat flux. In Eq. (43),
q and
q can be given by
. ,
4
0
4
0
T
S
c
q
T
S
c
q
m
s
m
s
x
x
(72) (72) where S0 is the solar radiation heat flux, and cs is the solar coefficient related to the shadow region, and
m
,
m
represent the effective solar absorptivity of the upper and lower surfaces, respectively, and
x
,
x represent the
albedo view factors of the upper and lower surfaces, respectively, and represents the Stefan-Boltzmann constant,
and
,
represent the emissivity of the upper and lower surfaces, respectively, and
T
T
T
r
,
T
T
T
r
represent the absolute temperature of the upper and lower surfaces, respectively. In case that the
plate is in a shadow region,
0
sc
. 6 Numerical simulations of satellite and solar array multibody system in thermal environment Thermal-dynamic analysis of the solar array is of vital importance to the safe operation of spacecraft. In this section,
we present two numerical examples to demonstrate the effectiveness of the proposed rigid-flexible-thermal
coupling model for the satellite and solar array multibody system in thermal environment. The first numerical
example focuses on comparing the different coupling models to reveal dynamic characteristics of thermally
induced vibration. The other numerical example is employed to analyze the deployment dynamic performance of
the solar array considering contact and thermal loading. 6.1 Numerical simulation of a satellite and solar array system in thermal environment 6.1 Numerical simulation of a satellite and solar array system in thermal environment Considering the influence of rigid body motion and elastic deformation on the albedo view factors of the
upper and lower surfaces, the albedo view factors are given by ,
2
/
0
2
/
0
cos
,
2
/
0
2
/
0
cos
x
x
(73) (73) where
and
are given by g
y
,
1
arccos
,
1
arccos
T
T
s
n
s
n
n
n
(74) (74) where the normal vectors are where the normal vectors are where the normal vectors are
y
x r
r
n
~
,
y
x r
r
n
~
. (75) (75) Since the
n and
n are closely related to the attitude motion of the rigid body as well as the elastic
deformation of the plate, the present formulation considers the influence of the attitude motion and the structural
deformation on the heat flux
q and
q , which is a complete rigid-flexible-thermal coupling model. However, in the conventional formulation,
and
are assumed to be constant due to the neglect of the
effects of rigid body motion and elastic deformation on the heat flux, and such model is not a complete
rigid-flexible-thermal coupling model. However, in the conventional formulation,
and
are assumed to be constant due to the neglect of the
effects of rigid body motion and elastic deformation on the heat flux, and such model is not a complete
rigid-flexible-thermal coupling model. As shown in Fig. 12, the flexible solar array is fully deployed, and its left edge is fixed to the rigid satellite. In
this simulation example, the rigid satellite is a cube, and the solar array is an isotropic rectangular flexible plate
with large deformation. 6.1 Numerical simulation of a satellite and solar array system in thermal environment The inertial reference frame O−XYZ is established at the centroid C of the rigid satellite,
and the geometric and material properties are given in Table 1. Defining the initial angel of incidence 0 as the solar angle, the coordinate vector of the solar sunlight is given
by
T
0
0
cos
0
sin
s
. Initially, the satellite is in static state, and the body fixed frame of the satellite is
parallel to the global coordinate system. The reference temperature is 273 K. The characteristic parameters of the
numerical method introduced in Section 4 are given by
8.0
, = 0.5. Table 1 Geometric and material properties. Table 1 Geometric and material properties. Satellite
Size of the rectangular satellite
2m×2m×2m
Mass
2000 Kg
Inertia matrix in body-fixed frame
Jx = Jy = Jz =200 Kg·m2
Solar array
Length
8 m
Width
1 m
Thickness
0.01 m
Elastic modulus
E = 1.93×109 Pa
Poisson’s ratio
= 0.3
Mass density
= 36.8 Kg/m3
Damping coefficient
= 0.0001 (1/s)
Thermal expansion coefficient
= 2.3×10-5 (1/K)
Specific heat of the plate
c = 921 J/(Kg·K)
Initial angle of incidence
Conductivity coefficient
k = 1.5 W/(m·K)
Solar heat flux
S0 = 1350 W/m2
Solar absorptivity of the plate
m = 0.79
Intensity of heat source
QT = 0
Stefan–Boltzmann constant
= 5.67×10-8 W/(m2·K4)
Emissivity of the upper surface
Emissivity of the lower surface
Gravitational acceleration
g = 0 6.1.1 Dynamic performance of on-orbit satellite and solar array system To reveal the advantages of rigid-flexible-thermal coupling model proposed in this paper for the satellite and solar
array system, two dynamic models are compared: the complete rigid-flexible-thermal coupling model (RFTC) and
the rigid-flexible coupling model considering thermal effect (RFCT). In the RFTC formulation,
and
are
closely related to
t
t
d
q
, therefore, the dynamic equations and the heat conductive equations are solved
simultaneously. However, in the RFCT formulation,
and
are assumed to be constant, which are equal to
0 and − 0, respectively. Since
t
t
T
F
is independent of
t
t
d
q
, the temperature results are not influenced by the
rigid body motion and the elastic deformation. 6.1 Numerical simulation of a satellite and solar array system in thermal environment For a more intuitive description of the deformation, the deflection is introduced. The deflection of an arbitrary
point on the plate can be written as
0
T
T
r
r
A
u
p
w
v
u
,
(76) (76) where r0 is the origin of the body-fixed frame of the plate, and Ap is the transformation matrix of the body-fixed
frame with respect to the inertial frame. where r0 is the origin of the body-fixed frame of the plate, and Ap is the transformation matrix of the body-fixed
frame with respect to the inertial frame. Time histories of the deflection of the corner point P in zc direction and the temperature gradient T1 =
z
T
/
of the corner point P are shown in Fig. 13(a) and (b), respectively. As can be seen in the figures, due to ignoring the
coupling effect of the solar heat flux, the rigid body motion and the elastic deformation, the RFCT model can not
reveal the thermally induced vibration. Accordingly, the vibration amplitude of the deflection is constant. On the
contrary, the vibration amplitudes of the deflection and the temperature gradient obtained by the RFTC model keep
growing, which explains the phenomenon of the thermally induced vibration effect. 0
50
100
150
200
-0.3
-0.2
-0.1
0.0
0.1
w [m]
Time [s]
Complete rigid-flexible-thermal coupling
Rigid-flexible coupling
0
50
100
150
200
0
30
60
90
120
150
180
dT/dz [K/m]
Time [s]
Complete rigid-flexible-thermal coupling
Rigid-flexible coupling
(a)
(b)
RFTC
RFCT
RFTC
RFCT
RFTC
RFCT
Fig. 13 Time histories of (a) deflection of point P in zc direction; (b) temperature gradient of point P in z direction 0
50
100
150
200
0
30
60
90
120
150
180
Time [s]
Complete rigid-flexible-thermal coupling
Rigid-flexible coupling
(b)
RFTC
RFCT
RFTC
RFCT 0
50
100
150
200
-0.3
-0.2
-0.1
0.0
0.1
w [m]
Time [s]
Complete rigid-flexible-thermal coupling
Rigid-flexible coupling
dT/dz [K/m]
(a)
RFTC
RFCT (a) (b) Fig. 13 Time histories of (a) deflection of point P in zc direction; (b) temperature gradient of point P in z direction In order to further analyze the effect of thermal load on the motion of the satellite, Fig. 14 gives the time
histories of the centroidal velocity of the satellite in z direction and the angular velocity of the satellite in yc
direction. 6.1 Numerical simulation of a satellite and solar array system in thermal environment Due to the coupling of the translational and the rotational motion of the satellite and the elastic
deformation of the solar array, the vibration amplitudes of the angular velocity and the centroidal velocity of the
satellite obtained by the RFTC model also keep growing. In a word, it reveals that the thermally induced vibration
phenomenon is obvious in practical engineering and needs to be paid attention to. -0.8
-0.4
0
0.4
0.8
1.2
x 10-4
0
50
100
150
200
vCz [m/s]
Time [s]
Complete rigid-flexible-thermal coupling
Rigid-flexible coupling
0
50
100
150
200
y [rad/s]
Time [s]
Complete rigid-flexible-thermal coupling
Rigid-flexible coupling
-0.6
-0.4
-0.2
0
0.2
0.4
0.6
0.8
x 10-3
(a)
(b)
RFTC
RFCT
RFTC
RFCT
Fig. 14 Time histories of (a) centroidal velocity of the satellite in z direction; (b) angular velocity of the satellite in yc direction 0
50
100
150
200
y
Time [s]
Complete rigid-flexible-thermal coupling
Rigid-flexible coupling
-0.6
-0.4
-0.2
0
0.2
0.4
0.6
0.8
x 10-3
(b)
RFTC
RFCT -0.8
-0.4
0
0.4
0.8
1.2
x 10
0
50
100
150
200
vCz [m/s]
Time [s]
Complete rigid-flexible-thermal coupling
Rigid-flexible coupling
y [rad/s]
(a)
RFTC
RFCT (b) (a) Fig. 14 Time histories of (a) centroidal velocity of the satellite in z direction; (b) angular velocity of the satellite in yc direction Fig. 14 Time histories of (a) centroidal velocity of the satellite in z direction; (b) angular velocity of the satellite in yc direction 6.1.2 Parameter analysis of thermally induced vibration In this section, based on the proposed rigid-flexible-thermal coupling model, some characteristic parameters are
investigated: the solar angle 0, specific heat c of the plate and the damping coefficient of plate on the thermally
induced vibration effect. According to the change of the solar angle, the time history of the angular velocity of the satellite in yc
direction is shown in Fig. 15(a). It is indicated that with the decrease of the solar angle, the average of the
deflection increases because of the increase of the intensity of the solar heat flux. However, the variation of the solar heat flux caused by the angular deformation of the plate and the rotation of the satellite becomes less
significant, therefore, the thermally induced vibration due to the rigid-flexible-thermal coupling effect also
decreases. Fig. 6.1 Numerical simulation of a satellite and solar array system in thermal environment 15(b) depicts the time history of the angular velocity of the satellite in yc direction at different plate
specific heats. As can be seen in the figure, due to the decrease of c and the increase of the thermal coefficient
)
/( c
k
, the thermally induced vibration also weakens. The time history of the angular velocity of the satellite in yc direction under different plate damping
coefficients is compared, as shown in Fig. 15(c). It can be obtained that with the increase of the structural damping
coefficient, the vibration amplitude of the deflection attenuates applied with the damping force, which indicates
that the increase of the structural damping coefficient contributes to weakening the thermally induced vibration. 0
50
100
150
200
-0.008
-0.004
0.000
0.004
0.008
y [rad/s]
Time [s]
0 = 65°
0 = 30°
0
50
100
150
200
-0.004
-0.002
0.000
0.002
0.004
y [rad/s]
Time [s]
c = 927
c = 100
0
50
100
150
200
-0.004
-0.002
0.000
0.002
0.004
y [rad/s]
Time [s]
= 0.0001
= 0.01
(a)
(b)
(c)
Fig. 15 Time histories of the angular velocity of the satellite in yc direction: under different (a) solar angles 0;
(b) plate specific heats c; (c) plate damping coefficients 0
50
100
150
200
-0.004
-0.002
0.000
0.002
0.004
y [rad/s]
Time [s]
= 0.0001
= 0.01 0
50
100
150
200
-0.004
-0.002
0.000
0.002
0.004
y [rad/s]
Time [s]
c = 927
c = 100
(b) 0
50
100
150
200
-0.008
-0.004
0.000
0.004
0.008
y [rad/s]
Time [s]
0 = 65°
0 = 30° (b) (b) (c) Fig. 15 Time histories of the angular velocity of the satellite in yc direction: under different (a) solar angles 0;
(b) plate specific heats c; (c) plate damping coefficients 6.1.3 Dynamic performance of manoeuvring satellite and solar array system In this section, the nonlinear dynamic performance of the manoeuvring satellite and solar array system applied with
thermal load is investigated. 6.1 Numerical simulation of a satellite and solar array system in thermal environment Additionally, the solar heat flux applied on the plate, and the satellite is applied with
translational constraint, which is given by xc = 0, yc = 0, zc = 0, and rotational constraint, which is given by x = 0,
z = 0, and z = 0, and
b
b
s
b
s
b
s
b
s
s
s
b
s
s
s
s
s
s
s
y
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
t
0
2
sin
2
0
2
sin
2
,
(77) (77) where tb = 200 s, ts = tb/8, s = /50, which is shown in Fig. 16. where tb = 200 s, ts = tb/8, s = /50, which is shown in Fig. 16. 0
50
100
150
200
250
300
-0.02
0.00
0.02
0.04
0.06
0.08
x [rad/s]
Time [s]
t = ts
t = tb
y
Fig. 16 Time history of y 0
50
100
150
200
250
300
-0.02
0.00
0.02
0.04
0.06
0.08
x [rad/s]
Time [s]
t = ts
t = tb
y Fig. 16 Time history of y On the basis of the RFTC model proposed in this paper, the deformed configurations of the satellite and solar
array multibody system as well as the distribution of the von Mises stress of the solar array at some time steps are
depicted in Fig. 17. Initially, the satellite rotates with angular velocity around the yc axis, which in turn drives the
motion of the solar array. In this situation, the solar array undergoes overall motion and large deformation. It is
shown that the stress concentration mainly occurs at the roots of the solar array, and the stress wave gradually
spreads toward the free edge of the solar array. Fig. 17 Motion of the satellite and solar array system at some time steps Fig. 6.1 Numerical simulation of a satellite and solar array system in thermal environment 17 Motion of the satellite and solar array system at some time steps For the purpose of highlighting the advantages of rigid-flexible-thermal coupling model for dynamic analysis
of the manoeuvring satellite and solar array system with overall motions and large deformation, three dynamic
models are compared: the RFTC model, the RFCT model and the rigid-flexible coupling model without
considering thermal effect (RFC). In the RFC formulation, only rigid-flexible coupling effects are considered,
while thermal effects are ignored. (b)
0
50
100
150
200
250
300
-400
-200
0
200
400
[
]
Time [s]
RFTC
RFCT
RFC
200
300
-334
-326 (a)
(b)
0
50
100
150
200
250
300
-0.4
-0.2
0.0
0.2
0.4
w [m]
Time [s]
RFTC
RFCT
RFC
0
50
100
150
200
250
300
-400
-200
0
200
400
dT/dz [K/m]
Time [s]
RFTC
RFCT
RFC
200
300
-334
-326
Fig. 18 Time histories of (a) deflection of point P in zc direction; (b) temperature gradient of point P in z direction (b) Fig. 18 Time histories of (a) deflection of point P in zc direction; (b) temperature gradient of point P in z direction Fig. 18 gives the time histories of the deflection w of the corner point P in zc direction and the temperature
gradient T1 =
z
T
/
of the corner point P, respectively. As can be seen in the figure, In the RFC model, except
for the acceleration and deceleration phases of y, the deflection of point P in zc direction maintains a small
amplitude vibration due to the rigid-flexible coupling effect. For t < 200s, the satellite undergoes periodic rotational motion in y direction, which leads to the periodic deflection in zc direction and the periodic variation of the
temperature gradient in thickness direction due to the complete rigid-flexible-thermal coupling effect, while the
deflection and the temperature gradient obtained by the RFCT model shows neither periodic variation nor
amplitude growing. Furthermore, it is shown that for t > 200 s, with the release of the applied angular velocity y,
the amplitude of vibration obtained by the RFTC model keeps growing slowly, which reveals the phenomenon of
the thermally induced vibration. To further analyze the thermal-dynamic performance of the manoeuvring satellite and solar array system, Fig. 19 gives the phase diagrams of point P in different models. 6.1 Numerical simulation of a satellite and solar array system in thermal environment From the phase diagram in the RFC model, it can be
seen that the trajectory of phase diagram exhibits a periodic motion without vibration under the action of y. In the
RFCT model, the trajectory of phase diagram can be viewed as the thermal vibrations superimposed on the basis of
the RFC model. Since the thermal loads are assumed to be constant in the RFCT model, the thermal vibrations
appear as low frequency oscillations. However, in the RFTC model, it shows the low frequency periodic motion as
well as the high frequency thermally induced vibration, due to the consideration of the influences of rigid body
motion and elastic deformation on the intensity of solar radiation. Therefore, it is necessary to adopt the more
precise rigid-flexible thermal coupling model for dynamic analysis of maneuvering satellite and solar array systems,
which can capture richer nonlinear characteristics. -10
-5
0
5
10
-0.6
-0.3
0.0
0.3
0.6
dx [m/s]
x [m]
-10
-5
0
5
10
-0.6
-0.3
0.0
0.3
0.6
dx [m/s]
x [m]
-10
-5
0
5
10
-0.6
-0.3
0.0
0.3
0.6
dx [m/s]
x [m]
RFTC
RFCT
RFC
(a)
(b)
(c)
START
ENT
START
ENT
START
ENT
Fig. 19 Phase diagrams point P in (a) RFTC model; (b) RFCT model; (c) RFC model
6.2 Deployment dynamics of a solar array in thermal environment with frictional contact
)
(t
d
Q
C
Zc
Yc
Xc
Solar array
Satellite
Sunlight
Guide rope
D1
D
)
(t
d
Rotational hinge
B
B1
Sunlight
P
O
X
Y
Z
D
B
P
Q
)
(t
d
①
②③
④
O
X
Z
(a)
(b)
Fig. 20 A simplified model of the deployment satellite and solar array system in thermal environment:
(a) partially deployed configuration; (b) stowed configuration -10
-5
0
5
10
-0.6
-0.3
0.0
0.3
0.6
dx [m/s]
x [m]
-10
-5
0
5
10
-0.6
-0.3
0.0
0.3
0.6
dx [m/s]
x [m]
-10
-5
0
5
10
-0.6
-0.3
0.0
0.3
0.6
dx [m/s]
x [m]
RFTC
RFCT
RFC
(a)
(b)
(c)
START
ENT
START
ENT
START
ENT
Fig. 6.1 Numerical simulation of a satellite and solar array system in thermal environment 19 Phase diagrams point P in (a) RFTC model; (b) RFCT model; (c) RFC model -10
-5
0
5
10
-0.6
-0.3
0.0
0.3
0.6
dx [m/s]
x [m]
RFC
(c)
START
ENT -10
-5
0
5
10
-0.6
-0.3
0.0
0.3
0.6
dx [m/s]
x [m]
dx [m/s]
RFCT
(b)
START
ENT -10
-5
0
5
10
-0.6
-0.3
0.0
0.3
0.6
dx [m/s]
x [m]
dx [m/s]
RFTC
(a)
START
ENT (b) Fig. 19 Phase diagrams point P in (a) RFTC model; (b) RFCT model; (c) RFC model 6.2 Deployment dynamics of a solar array in thermal environment with frictional cont eployment dynamics of a solar array in thermal environment with frictional contact )
(t
d
Q
C
Zc
Yc
Xc
Solar array
Satellite
Sunlight
Guide rope
D1
D
)
(t
d
Rotational hinge
B
B1
Sunlight
P
O
X
Y
Z
D
B
P
Q
)
(t
d
①
②③
④
O
X
Z
(a)
(b)
Fig. 20 A simplified model of the deployment satellite and solar array system in thermal environment:
(a) partially deployed configuration; (b) stowed configuration )
(t
d
Q
C
Zc
Yc
Xc
Solar array
Satellite
Sunlight
Guide rope
D1
D
)
(t
d
Rotational hinge
B
B1
Sunlight
P
O
X
Y
Z
(a) Sunlight
D
B
P
Q
)
(t
d
①
②③
④
O
X
Z
(b) Satellite Sunlight Rotational hinge Rotational hinge (a) (b) Fig. 20 A simplified model of the deployment satellite and solar array system in thermal environment:
(a) partially deployed configuration; (b) stowed configuration The second example deals with the deployment of the solar array in thermal environment considering frictional
contact. The simplified model of the satellite and solar array system in thermal environment is illustrated by Fig. 20,
which consists of four guide wires, eight solar panels and eight rotational hinges. It is worth mentioning that the
translational and attitude motion of the satellite is controlled during the deployment of the solar array, and it can be
regarded as points B, D, B1 and D1 are restricted to move on the guide wires ignoring the deformation of the guide wires. 6.1 Numerical simulation of a satellite and solar array system in thermal environment Since the direction of sunlight is along the negative z axis, and the solar array are symmetrically distributed
about the central satellite, it is sufficient to analyze the deployment of the solar panels on one side. O−XYZ is the
inertial reference frame, which is consistent with the body-fixed frame of the satellite C−XcYcZc. The length of one side solar array is LP = 5 m, and the solar angle 0 = 0, and the other geometric and material
properties refer to Table 1. The initial angle between the adjacent solar panels is 0 = 2o, as shown in Fig. 20(b). Considering the contact of solar panels and the contact between solar panel and satellite, it requires the STS and
STRS contact elements. The normal and tangential penalty parameters for the STS and STRS contact elements are
N = T = 105 N/m2, and the static and dynamic frictional coefficients are s = d = 0.3. In addtion, the points D and
D1 are subjected to a prescribed translational displacement vector
T]
0
0
)
(
[
)
(
t
d
t
d
m. The piecewise function is
e
P
d
P
d
d
d
t
t
t
L
t
t
t
t
t
a
L
t
t
t
t
t
t
a
t
a
t
t
t
a
t
d
3
3
2
2
3
2
1
1
1
2
1
1
2
)
(
5.0
)
(
5.0
0
5.0
)
(
,
(78) (78) where t1 = 12 s, t2 = 108 s, t3 = 120 s and te = 200 s. ad = LP/t1t2 ensures that the deployment of the solar array ends
with a fully deployed configuration, and the max velocity of the drive is defined as vm = vc = ad t1. Based on the complete rigid-flexible-thermal coupling model proposed in this paper, the changing of
configurations at some time steps are shown in Fig. 21, and the color maps represent the distribution of the von
Mises stress. The initial stress of the folded solar panels is set to zero, and then the solar panels are gradually
deployed applied with the driving constraints. 6.1 Numerical simulation of a satellite and solar array system in thermal environment 22 Time history of temperature gradients of points P, B and Q (see Fig. 20) in z direction 6.2.1 Influence of contact 6.2.1 Influence of contact 6.2.1 Influence of contact In order to illustrate the necessity of contact, two models are employed to verify. The first one considers the
integration of the STS and STRS contact elements, and the other does not consider contact. For the sake of
intuitiveness, the configuration of the satellite and solar array system at t = 30 s is selected for analysis, as shown in
Fig. 23. It can be seen in Fig. 23(a) that the solar array can deploy properly without over-penetration in the
consideration of contact. On the contrary, it is clearly observed that the large penetration appears not only between
the solar panels, but also between the satellite and the solar panels in case that the contact is not taken into account. In conclusion, for the deployment of the satellite and solar array system, it requires a combination of STS and
STRS contact elements to avoid mutual penetration, thereby ensuring computation accuracy. Fig. 23 Configurations of the (a) contact model; (b) no contact model at t = 30 s Fig. 23 Configurations of the (a) contact model; (b) no contact model at t = 30 s With the view of further analyzing the effect of contact, Fig. 24 depicts the time history of coordinate x of
points B and P obtained by the contact and no contact models. It can be found that the values of the coordinate x of
points B and P are less than 1, that is, the solar panels penetrate into the satellite, which violates the real situation. After the contact no longer occurs, the curves of the two contact models gradually tend to be consistent, and the
solar array is fully deployed at t = 120 s. Therefore, we can conclude that the contact affects not only the motions of
the system, but also the shadow regions, which is related to the intensity of the solar radiation. 6.1 Numerical simulation of a satellite and solar array system in thermal environment Because of contact, it can be found that the stress of panels 1 and 2 is
greater than that of panels 3 and 4 at t = 30 s and t = 60 s. As the thermal loads continue, the panels 3 and 4
experience greater thermal stress, which can be seen in the configurations at t = 90 s and t = 120 s. After the solar
array is fully deployed, we can observe that the stress distribution of the four solar panels is relatively balanced,
accompanied by obvious thermal deflection. Fig. 21 The configurations and the von Mises stress of the satellite and solar array system at some time steps Fig. 21 The configurations and the von Mises stress of the satellite and solar array system at some time steps Fig. 22 presents the time history of temperature gradients of the points P, B and Q during the deployment of
the solar array, and these test points can comprehensively reflect the heating conditions of the solar array. It can be
seen that for t < 50 s, the temperature gradient at point P has a zero value, which is due to the fact that point P is in
the shadow region at the time of contact and cannot receive solar radiation. Since one of the plates connecting the
point B is in the shadow region at the initial stage, the temperature gradient rate at point B is lower than that at point
Q. It is the reason that the panels 3 and 4 experience greater thermal stress than panels 3 and 4 during the
deployment of the solar array. Finally, the solar panels are fully deployed, and the temperature gradients of the
three points are the same. Therefore, the influence of contact and thermal loads on the deployment process of the
solar array is essential, which needs to be paid more attention to in engineering. 0
50
100
150
200
0
50
100
150
200
250
300
350
400
dT/dz [K/m]
Time [s]
P
B
Q
Shadow region
0
50
25
0
50
25
350 K/m
Fig. 22 Time history of temperature gradients of points P, B and Q (see Fig. 20) in z direction 0
50
100
150
200
0
50
100
150
200
250
300
350
400
dT/dz [K/m]
Time [s]
P
B
Q
Shadow region
0
50
25
0
50
25
350 K/m Fig. 6.2.2 Influence of thermal load Since the thermal loads have a significant effect on the deployment of the solar array, two different thermal models
are compared and analyzed in this section. Fig. 25(a) presents the time history of coordinates z of point Q obtained
by the thermal and no thermal models, respectively. By observing the coordinates zQ of the two models, it can be
seen that when the solar panels 3 and 4 are fully deployed, the thermal model excites high frequency oscillations with gradually decreasing amplitudes. Combined with the configuration at t = 55 s in Fig. 25(b), it can be seen that
since point B of the no thermal model moves later than the thermal model, point Q has a greater vertical velocity as
it passes the horizontal position, resulting in a greater offset magnitude. In addition, the final configurations of the
thermal and no thermal models at t = 200 s are also presented in Fig. 25(b), in which we can observe that obvious
deflections of the thermal model occur at points P and Q due to the effects of the boundary constraints and the
thermal loads, which cannot be obtained by the no thermal model. Fig. 24 Time history of coordinates x of points B and P in different contact models Fig. 24 Time history of coordinates x of points B and P in different contact models 0
50
100
150
200
-1.5
-1.2
-0.9
-0.6
-0.3
0.0
0.3
Coordinate z [m]
Time [s]
zQ (Thermal)
zQ (No thermal)
z = 0
t = 55 s
t = 200 s
(a)
(b)
Fig. 25 (a) Time history of coordinates z of point Q in different thermal models;
(b) Configurations of different thermal models at t = 55 s and t = 200 s (b) (b) Fig. 25 (a) Time history of coordinates z of point Q in different thermal models;
(b) Configurations of different thermal models at t = 55 s and t = 200 s To further analyze the thermal expansion and deflection, the time history of deflection of point P in different
models is given in Fig. 26. Here, u, v and w denote the deflection of point P in the x, y and z directions, respectively. The values of u and v remain zero in the no thermal model, but eventually exhibit positive expansion in the thermal
model. 6.2.2 Influence of thermal load By comparing the curve of w in the two models, it can be observed that when the solar array is fully
deployed at t = 120 s, the vibration amplitude of the thermal model is greater than that of the no thermal model, and
the decay time is longer. From the results in the figure, it is necessary to pay attention to the phenomenon of
thermal deflection during the deployment of solar array. 6.2.3 Comparison of different drive speeds 26 Time history of deflection of point P in different
thermal models (b)
0
50
100
150
200
-0.03
-0.02
-0.01
0.00
0.01
0.02
0.03
0.04
vBx [m/s]
Time [s]
vm = vc
vm = 5vc /6
vm = 3vc /4 Fig. 28 Time histories of (a) displacement; (b) velocity of point B in the x direction under different cases Fig. 28 plots the dynamic response of point B at three different drive speeds. It can be seen in Fig. 28(a) that
the deployment time of the solar array of the three cases are significantly different, and the changes of xB are
relatively stable. From the results in Fig. 28(b), we can observe that there are two obvious high frequency
vibrations during the deployment of the solar array. The first occurs when the solar panels 3 and 4 are fully
deployed, the other occurs when the solar panels 1 and 2 are fully deployed. Since the thermal effect is positively
related to action time of the thermal load, the thermal stress increases with decreasing drive speed, resulting in a
greater amplitude of oscillation, especially when the plates are fully deployed. 6.2.3 Comparison of different drive speeds In engineering, the impact of drive speed on the satellite and solar array system is another concerned issue. In this
section, we focus on comparing the results of three different drive speeds acting on the solar array in a thermal
environment. As illustrated in Fig. 20, the points D and D1 are subjected to a prescribed translational displacement along the x direction. As shown in Fig. 27, the drive speed curves in different cases have the same displacement at t
= te, in which the black curve represents the basic model, and its expression refers to Eq. (78). -0.001
0.000
0.001
-0.01
0.00
0.01
0
50
100
150
200
-0.006
-0.004
-0.002
0.000
u [m]
Heat
No heat
v [m]
w [m]
Time [s]
110
160
-5.8
-5.0
x10-3
110
180
-2.5
2.5
x10-4
Thermal
No thermal
Drive speed v [m/s]
Time [s]
vm = vc
vm = 5vc /6
vm = 3vc /4
t1
t2 t3
te
vc
5vc/6
t2
5
6
t2
3
4
t1
6
5
t1
4
3
3vc/4
Fig. 26 Time history of deflection of point P in different
thermal models
Fig. 27 Drive speed curves in three cases
(a)
(b)
0
50
100
150
200
-0.03
-0.02
-0.01
0.00
0.01
0.02
0.03
0.04
vBx [m/s]
Time [s]
vm = vc
vm = 5vc /6
vm = 3vc /4
0
50
100
150
200
1.0
1.2
1.4
1.6
1.8
2.0
2.2
2.4
xB [m]
Time [s]
vm = vc
vm = 5vc /6
vm = 3vc /4
Fig. 28 Time histories of (a) displacement; (b) velocity of point B in the x direction under different cases -0.001
0.000
0.001
-0.01
0.00
0.01
0
50
100
150
200
-0.006
-0.004
-0.002
0.000
u [m]
Heat
No heat
v [m]
w [m]
Time [s]
110
160
-5.8
-5.0
x10-3
110
180
-2.5
2.5
x10-4
Thermal
No thermal Drive speed v [m/s]
Time [s]
vm = vc
vm = 5vc /6
vm = 3vc /4
t1
t2 t3
te
vc
5vc/6
t2
5
6
t2
3
4
t1
6
5
t1
4
3
3vc/4
Fig. 27 Drive speed curves in three cases Drive speed v [m/s]
5
3 Fig. 27 Drive speed curves in three cases Fig. 27 Drive speed curves in three cases Fig. 7 Conclusions A strong rigid-flexible-thermal coupling dynamic model for hub and multi-plate multibody system with large
deformation and frictional contact is proposed. The temperature field is expressed with Taylor polynomials to gain
two-dimensional heat conduction equations, and a thermal integrated ANCF thin plate element is developed. Accounting for the influences of the attitude motion and structural deformation on the intensity of the solar
radiation, the relation between the heat flux and the absolute coordinates is derived. To address the contact problem, STS and STRS frictional contact elements are developed using the penalty method and the Coulomb friction law
with a penalty regularization, which can effectively avoid the mutual over penetration and capture the stick-slip
behavior. Subsequently, the rigid-flexible-thermal coupling equations, which are combined of the heat conduction
equations and the Lagrange dynamic equations of the first kind, are derived. Furthermore, a new numerical method
combining the modified central difference method and the generalized- method is proposed to solve the strong
rigid-flexible-thermal coupling equations. By setting comparative tests in ABAQUS, the correctness of the proposed thermal-structural coupling model
is validated, and the necessary of the geometric nonlinearity is proved. Moreover, the advantages of the proposed
model in capturing thermally induced vibration is revealed. In order to further demonstrate the effectiveness of the
proposed rigid-flexible-thermal coupling method in hub and multi-plate system, two numerical examples of the
satellite and solar array multibody system are conducted. The first example compares the results of three dynamic
models: RFTC model, RFCT model and RFC model. Due to the consideration of the elastic deformation as well as
the rigid body motion on the intensity of heat flux, RFTC model can reveal the thermally induced vibration effect,
which cannot be achieved by the other two models. Parameter analysis illustrates that the thermally induced
vibration effect can be reduced through the decrease of the solar angle and the specific heat or the increase of the
damping coefficient. Then, the nonlinear coupling behavior of the manoeuvring satellite and solar array system can
be revealed by the proposed RFTC model. The second example is employed to analyze the deployment dynamic
performance of the solar array in thermal environment with frictional contact. It is concluded that the contact
affects not only the motions of the system, but also the shadow regions, which is related to the intensity of the solar
radiation. 7 Conclusions Due to the effects of the boundary constraints and the thermal loads, obvious thermal deflection and high
frequency vibration can be induced, especially when the plates are fully deployed. Since the thermal effect is
positively related to action time of the thermal load, the amplitude of thermal vibration increases with the
decreasing of the drive speed. Acknowledgements This research was supported by the General Program (No.12272221) of the National Natural
Science Foundation of China, the Key Program (No.11932001) of the National Natural Science Foundation of China, for
which the authors are grateful. This research was also supported by the Key Laboratory of Hydrodynamics (Ministry of
Education). Declaration on conflict of interest The authors declare that they have no conflicts of interest to this work. Declaration on conflict of interest The authors declare that they have no conflicts of interest to this work. References 1. Thornton, E.A., Kim, Y.A.: Thermally induced bending vibrations of a flexible rolled-up solar array. J. Spacecr. Rockets. 30(4), 438-448 (1993) 1. Thornton, E.A., Kim, Y.A.: Thermally induced bending vibrations of a flexible rolled-up solar array. J. Spacecr. Rockets. 30(4), 438-448 (1993) 1. Thornton, E.A., Kim, Y.A.: Thermally induced bending vibrations of a flexible rolled up solar array. J. Spacecr. Rockets. 30(4), 438-448 (1993)
2. Johnston, J.D., Thornton, E.A.: Thermally induced attitude dynamics of a spacecraft with a flexible appendage. J. Guid. Control Dynam. 21(4), 581-587 (1998) 2. Johnston, J.D., Thornton, E.A.: Thermally induced attitude dynamics of a spacecraft with a flexible appendage. J. Guid. Control Dynam. 21(4), 581-587 (1998) 3. Johnston, J.D., Thornton, E.A.: Thermally induced dynamics of satellite solar panels. J. Spacecr. Rockets. 37(5),
604-613 (2000) 4. Fan, W., Liu, J.: Geometric nonlinear formulation for thermal-rigid-flexible coupling system. Acta Mech. Sin. 29(5),
728-737 (2013) 5. Shen, Z., Tian, Q., Liu, X., Hu, G.: Thermally induced vibrations of flexible beams using Absolute Nodal Coordinate
Formulation. Aerosp. Sci. Technol. 29(1), 386-393 (2013) 6. Shen, Z., Hu, G.: Thermally Induced Dynamics of a Spinning Spacecraft with an Axial Flexible Boom. J. Spacecr. Rockets. 52, 1503-1508 (2015) Hu, G.: Thermoelastic–structural analysis of space thin-walled beam under solar flux. AIAA J. 57
5 (2019) 8. Čepon, G., Starc, B., Zupančič, B., Boltežar, M.: Coupled thermo-structural analysis of a bimetallic strip using the
absolute nodal coordinate formulation. Multibody Syst. Dyn. 41(4), 391-402 (2017) 9. Liu, L., Sun, S., Cao, D., Liu, X.: Thermal-structural analysis for flexible spacecraft with single or double solar
panels: A comparison study. Acta Astronaut. 154, 33-43 (2019) 10. Liu, L., Cao, D., Huang, H., Shao, C., Xu, Y.: Thermal-structural analysis for an attitude maneuvering flexible
spacecraft under solar radiation. Int. J. Mech. Sci. 126, 161-170 (2017) 11. Liu, L., Wang, X., Sun, S., Cao, D., Liu, X.: Dynamic characteristics of flexible spacecraft with double solar panels
subjected to solar radiation. Int. J. Mech. Sci. 151, 22-32 (2019) 12. Li, Y., Wang, C., Huang, W.: Rigid-flexible-thermal analysis of planar composite solar array with clearance joint
considering torsional spring, latch mechanism and attitude controller. Nonlinear Dyn. 96(3), 2031-2053 (2019) 13. Liu, J., Pan, K.: Rigid-flexible-thermal coupling dynamic formulation for satellite and plate m
Aerosp. Sci. Technol. 52, 102-114 (2016) 14. Shabana, A.A.: Flexible multibody dynamics: review of past and recent developments. Multibody Syst. Dyn. 1(2),
189-222 (1997) 15. References Dmitrochenko, O.N., Pogorelov, D.Y.: Generalization of plate finite elements for absolute nodal coordinate
formulation. Multibody Syst. Dyn. 10(1), 17-43 (2003) 16. Dmitrochenko, O., Mikkola, A.: Two simple triangular plate elements based on the absolute nodal coordinate
formulation. J. Comput. Nonlin. Dyn. 3(4), 041012 (2008) 17. Dufva, K., Shabana, A.A.: Analysis of thin plate structures using the absolute nodal coordinate formulation. P. I. Mech. Eng. K-J Mul. 219(4), 345-355 (2005) 18. Ren, H.: Fast and robust full-quadrature triangular elements for thin plates/shells with large deformations and large
rotations. J. Comput. Nonlin. Dyn. 10(5), 051018 (2015) 19. Schwab, A.L., Gerstmayr, J., Meijaard, J.P.: Comparison of three-dimensional flexible thin plate elements for
multibody dynamic analysis: finite element formulation and absolute nodal coordinate formulation. In: ASME 2007
International Design Engineering Technical Conferences and Computers and Information in Engineering Conference,
Las Vegas, Nevada, USA (2007) 20. Vaziri Sereshk, M., Salimi, M.: Comparison of finite element method based on nodal displacement and absolute
nodal coordinate formulation (ANCF) in thin shell analysis. Int. J. Numer. Meth. Bio. 27(8), 1185-1198 (2011) 21. Hyldahl, P., Mikkola, A.M., Balling, O., Sopanen, J.T.: Behavior of thin rectangular ANCF shell elements in various
mesh configurations. Nonlinear Dyn. 78(2), 1277-1291 (2014) 22. Liu, C., Tian, Q., Hu, H.: New spatial curved beam and cylindrical shell elements of gradient-deficient Absolute
Nodal Coordinate Formulation. Nonlinear Dyn. 70(3), 1903-1918 (2012) 23. Gerstmayr, J., Sugiyama, H., Mikkola, A.: Review on the Absolute Nodal Coordinate Formulation for Large
Deformation Analysis of Multibody Systems. J. Comput. Nonlin. Dyn. 8(3), 031016 (2013) 24. Liu, C., Tian, Q., Yan, D., Hu, H.: Dynamic analysis of membrane systems undergoing overall motions, large
deformations and wrinkles via thin shell elements of ANCF. Comput. Meth. Appl. Mech. Eng. 258, 81-95 (2013) 24. Liu, C., Tian, Q., Yan, D., Hu, H.: Dynamic analysis of membrane systems undergoing overall motions, large
deformations and wrinkles via thin shell elements of ANCF. Comput. Meth. Appl. Mech. Eng. 258, 81-95 (2013)
25. Shen, Z., Hu, G.: Thermally induced vibrations of solar panel and their coupling with satellite. Int. J. Appl. Mech. 05, 25. Shen, Z., Hu, G.: Thermally induced vibrations of solar panel and their coupling with satellite. Int. J. Appl. Mech. 05,
1350031 (2013) Z., Lan, P.: A novel method of thermo-mechanical coupled analysis based on the unified description. . Theory. 134, 376-392 (2019) 26. References Cui, Y., Yu, Z., Lan, P.: A novel method of thermo-mechanical coupled analysis based on the un
Mech. Mach. Theory. 134, 376-392 (2019) 27. Cui, Y., Lan, P., Zhou, H., Yu, Z.: The Rigid-Flexible-Thermal Coupled Analysis for Spacecraft Carrying
Large-Aperture Paraboloid Antenna. J. Comput. Nonlin. Dyn. 15(3) (2020) 28. Wriggers, P.: Computational Contact Mechanics. Springer, Berlin,Wiley (2006) 28. Wriggers, P.: Computational Contact Mechanics. Springer, Berlin,Wiley (2006) 29. Konyukhov, A., Izi, R.: Introduction to computational contact mechanics: a geometrical approach. W
(2015) Lankarani, H.M.: Contact Force Models for Multibody Dynamics, In: Solid Mechanics and Its
. Springer, Berlin (2016) 30. Flores, P., Lankarani, H.M.: Contact Force Models for Multibody Dynamics, In: Solid Me
Applications. Springer, Berlin (2016) 31. Flores, P.: Contact mechanics for dynamical systems: a comprehensive review. Multibody Syst. Dyn. 54(2), 127-177
(2022) 32. Konyukhov, A., Schweizerhof, K.: Computational Contact Mechanics: Geometrically Exact Theory for Arbitrary
Shaped Bodies. Springer, Berlin (2013) 33. Schweizerhof, K., Konyukhov, A.: Covariant description for frictional contact problems. Comput. Mech. 35, 190-213
(2005) 34. Yu, L., Zhao, Z., Tang, J., Ren, G.: Integration of absolute nodal elements into multibody system. Nonlinear Dyn. 62(4), 931-943 (2010) 35. Shi, J., Liu, Z., Hong, J.: Dynamic contact model of shell for multibody system applications. Multibody Syst. Dyn. 44(4), 335-366 (2018) 36. Sun, D., Liu, C., Hu, H.: Dynamic computation of 2D segment-to-segment frictionless conta
multibody system subject to large deformation. Mech. Mach. Theory. 140, 350-376 (2019) 36. Sun, D., Liu, C., Hu, H.: Dynamic computation of 2D segment-to-segment frictionless contact for a flexible
multibody system subject to large deformation. Mech. Mach. Theory. 140, 350-376 (2019)
37 Sun D Liu C Hu H : Dynamic computation of 2D segment to segment frictional contact for a flexible multibody 37. Sun, D., Liu, C., Hu, H.: Dynamic computation of 2D segment-to-segment frictional contact for a fl
system subject to large deformations. Mech. Mach. Theory. 158, 104197 (2021) 38. Gay Neto, A., Wriggers, P.: Master-master frictional contact and applications for beam-shell interaction. Comput. Mech. 66(6), 1213-1235 (2020) 39. Tang, L., Liu, J.: Frictional contact analysis of sliding joints with clearances between flexible beams and rigid holes
in flexible multibody systems. Multibody Syst. Dyn. 49(2), 155-179 (2019) 40. Lei, B., Ma, Z., Liu, J., Liu, C.: Dynamic modelling and analysis for a flexible brush sampling mechanism. Multibody
Syst. Dyn. 56(4), 335-365 (2022) 41. References Yuan, T., Liu, Z., Zhou, Y., Liu, J.: Dynamic modeling for foldable origami space membrane structure with
contact-impact during deployment. Multibody Syst. Dyn. 50(1), 1-24 (2020) 42. Yuan, T., Tang, L., Liu, Z., Liu, J.: Nonlinear dynamic formulation for flexible origami-based deployable structures
considering self-contact and friction. Nonlinear Dyn. 106(3), 1789-1822 (2021) y
Computer Aided Kinematics and Dynamics of Mechanical Systems. Allyn and Bacon, Boston (1989) 43. Haug, E.J.: Computer Aided Kinematics and Dynamics of Mechanical Systems. Allyn and Bacon, B 44. Hong, J.: Computational dynamics of multibody systems. Higher Education Press, Beijing (1999) 45. Tang, L., Liu, J.: Modeling and analysis of sliding joints with clearances in flexible multibody systems. Nonlinear
Dyn. 94(4), 2423-2440 (2018) 46. Liu, Z., Liu, J.: Experimental validation of rigid-flexible coupling dynamic formulation for hub–beam system. Multibody Syst. Dyn. 40(3), 303-326 (2017) 47.Arnold, M., Brüls, O.: Convergence of the generalized-α scheme for constrained mechanical systems. Multibody Syst. Dyn. 18(2), 185-202 (2007) 48. Noels, L., Stainier, L., Ponthot, J.P.: Self-adapting time integration management in crash-worthiness and sheet metal
forming computations. Int. J. Vehicle Des. 30(1-2), 67-114 (2002) 48. Noels, L., Stainier, L., Ponthot, J.P.: Self-adapting time integration management in crash-worthiness and sheet metal
forming computations. Int. J. Vehicle Des. 30(1-2), 67-114 (2002)
|
https://openalex.org/W2096714740
|
https://europepmc.org/articles/pmc6269888?pdf=render
|
English
| null |
Thymoquinone Induces Mitochondria-Mediated Apoptosis in Acute Lymphoblastic Leukaemia in Vitro
|
Molecules/Molecules online/Molecules annual
| 2,013
|
cc-by
| 8,619
|
Molecules 2013, 18, 11219-11240; doi:10.3390/molecules180911219
molecules
ISSN 1420-3049
www.mdpi.com/journal/molecules
Article
Thymoquinone Induces Mitochondria-Mediated Apoptosis in
Acute Lymphoblastic Leukaemia in Vitro
Landa Zeenelabdin Ali Salim 1, Syam Mohan 1,*, Rozana Othman 1, Siddig Ibrahim Abdelwahab 2,
Behnam Kamalidehghan 1, Bassem Y. Sheikh 3 and Mohamed Yousif Ibrahim 1
1 Department of Pharmacy, Faculty of Medicine, University of Malaya, KualaLumpur 50603, Malaysia
2 Medical Research Centre, Jazan University, P.O. Box 114 Jazan, Jazan 45142, Saudi Arabia
3 College of Medicine, Taibah University, Al Madinah Al Monawarah 30001, Saudi Arabia
* Author to whom correspondence should be addressed; E-Mail: syammohanm@yahoo.com;
Tel.: +603-7967-7520; Fax: +603-7967-4964.
Received: 18 July 2013; in revised form: 25 August 2013 / Accepted: 30 August 2013 /
Published: 12 September 2013
OPEN ACCESS Molecules 2013, 18, 11219-11240; doi:10.3390/molecules180911219 Keywords: anticancer; apoptosis; mitochondria; thymoquinone; CEMss 1. Introduction Nigella sativa, also known as black cumin (black seed), is an annual herbaceous plant belonging to
the Ranunculacea family which grows in countries bordering the Mediterranean Sea and India [1]. It
has been used as traditional herbal medicine for more than 2,000 years [2]. Apart from this
complimentary medicinal use, it has been used in Asia, Middle East and Africa as a medicinal food to
support health and fight many diseases [3]. N. sativa is one of the most extensively studied plants [4]. It used as a food additive and flavour worldwide [2,5]. They are also used as a natural remedy for
asthma, hypertension, diabetes, inflammation, cough, bronchitis, headache, eczema, fever, dizziness
and influenza [4,6]. The seeds are known to be carminative, stimulant, diuretic, emenagouge and
galactogogue, and have been used in treating fever [7]. The biological activity of N. sativa seeds is
attributed to their essential oil components [8]. The main compounds contained are thymoquinone
(30%–48%), p-cymene (7%–15%), carvacrol (6%–12%), 4-terpineol (2%–7%), t-anethole (1%–4%)
and the sesquiterpene longifolene (1%–8%) [9]. In Malaysia, N. sativa is popularly known as “el Habbatus el Sauda”. Currently, the black seed oil
has been widely commercialized and due to its amazing healing power, numerous research projects are
being carried out from time to time to improve its quality [10,11]. Thymoquinone (TQ, 2-isopropyl-5-
methyl-1,4-benzoquinone, Figure 1b) is the major component and the most bioactive constituent of the
volatile oil of this seed, which has been shown to possess anti-inflammatory, antioxidant and
anti-carcinoma effects [12]. In recent years, TQ and its effects on different cancer cell lines have been
widely studied; these effects include inhibition of cancer cell viability. In almost all pancreatic cancer
cell lines tested, the inhibition was up to 70% [13]. Anti-proliferative and pro-apoptotic activities of
TQ in both NSCLC and SCLC cell line [14]. Breast cancer cell lines [15] and liver cell lines [16] have
also been studied, including in some animal models [17]. The problem of cancer burden in Malaysia is growing, as in other countries around the World, since
it is one of the major health problems, which leads to death [18]. Leukaemia is one of the most
common childhood cancers worldwide and also in Malaysia [19]. Childhood leukaemia represents 4.7%
of childhood cancers in the Malaysian population [20]. This is treated by multidisciplinary efforts
including chemotherapy. Landa Zeenelabdin Ali Salim 1, Syam Mohan 1,*, Rozana Othman 1, Siddig Ibrahim Abdelwahab 2,
Behnam Kamalidehghan 1, Bassem Y. Sheikh 3 and Mohamed Yousif Ibrahim 1 1 Department of Pharmacy, Faculty of Medicine, University of Malaya, KualaLumpur 50603, Malaysia
2 Medical Research Centre, Jazan University, P.O. Box 114 Jazan, Jazan 45142, Saudi Arabia
3 College of Medicine, Taibah University, Al Madinah Al Monawarah 30001, Saudi Arabia Department of Pharmacy, Faculty of Medicine, University of Malaya, KualaLumpur 50603, Malaysia
2 Medical Research Centre, Jazan University, P.O. Box 114 Jazan, Jazan 45142, Saudi Arabia
3 College of Medicine, Taibah University, Al Madinah Al Monawarah 30001, Saudi Arabia ,
y,
,
,
3 College of Medicine, Taibah University, Al Madinah Al Monawarah 30001, Saudi Arabia * Author to whom correspondence should be addressed; E-Mail: syammohanm@yahoo.com;
Tel.: +603-7967-7520; Fax: +603-7967-4964. * Author to whom correspondence should be addressed; E-Mail: syammohanm@yahoo.com;
Tel.: +603-7967-7520; Fax: +603-7967-4964. Received: 18 July 2013; in revised form: 25 August 2013 / Accepted: 30 August 2013 /
Published: 12 September 2013 Received: 18 July 2013; in revised form: 25 August 2013 / Accepted: 30 August 2013 /
Published: 12 September 2013 Abstract: There has been a growing interest in naturally occurring compounds from
traditional medicine with anti-cancer potential. Nigella sativa (black seed) is one of the
most widely studied plants. This annual herb grows in countries bordering the
Mediterranean Sea and India. Thymoquinone (TQ) is an active ingredient isolated from
Nigella sativa. The anti-cancer effect of TQ, via the induction of apoptosis resulting from
mitochondrial dysfunction, was assessed in an acute lymphocyte leukemic cell line
(CEMss) with an IC50 of 1.5 µg/mL. A significant increase in chromatin condensation in
the cell nucleus was observed using fluorescence analysis. The apoptosis was then
confirmed by Annexin V and an increased number of cellular DNA breaks in treated cells
were observed as a DNA ladder. Treatment of CEMss cells with TQ encouraged apoptosis
with cell death-transducing signals by a down-regulation of Bcl-2 and up-regulation of
Bax. Moreover, the significant generation of cellular ROS, HSP70 and activation of
caspases 3 and 8 were also observed in the treated cells. The mitochondrial apoptosis was
clearly associated with the S phase cell cycle arrest. In conclusion, the results from the
current study indicated that TQ could be a promising agent for the treatment of leukemia. Molecules 2013, 18 Molecules 2013, 18 11220 1. Introduction One of the main problems of the present chemotherapy in treating tumour
patients is the toxicity of the drugs used. Most of the existing anticancer drugs, unfortunately, also
attack normal proliferating cells [21]. Therefore, being generally considered as safe, consumption of
natural compounds is currently a major interest in health care. In the current study, we evaluated the
potential of TQ on T acute lymphoblastic leukaemia using CEMss cells as an in vitro model. 2.1. Results 2.1.1. Cell Culture and Viability Assay 2. Results and Discussion 2.1. Results 2.1.1. Cell Culture and Viability Assay The effects of TQ on the viability of CEMss cells were measured using the MTT assay. Cellular
proliferation following 24 h of exposure to TQ showed significant inhibition in TQ-treated cells compared Molecules 2013, 18 11221 to non-treated cells (controls). As shown in Figure 1, the IC50 of TQ was 1.5 ± 0.04 μg/mL following 24 h
of treatment. The proliferation of TQ-treated cells decreased as the TQ concentration increased. Figure 1. Effects of Thymoquinone on cell viability in CEMss. (a) The cell viability of
cells after 24 h of treatment. Each point is the mean ± S.D. of three independent
experiments. (b) The chemical structure of thymoquinone. Figure 1. Effects of Thymoquinone on cell viability in CEMss. (a) The cell viability of
cells after 24 h of treatment. Each point is the mean ± S.D. of three independent
experiments. (b) The chemical structure of thymoquinone. 2.1.2. Quantification of Apoptosis Using Propidium Iodide and Acridine Orange Double-Staining
0
10
20
30
40
50
60
70
80
90
100
110
0
50
100
150
% Inhibition
Concentration (microgram/mL)
a
b
2-isopropyl-5-methyl-1, 4
benzoquinone 0
10
20
30
40
50
60
70
80
90
100
110
0
50
100
150
% Inhibition
Concentration (microgram/mL)
a
b
2-isopropyl-5-methyl-1, 4
benzoquinone b b a 2-isopropyl-5-methyl-1, 4
benzoquinone 2.1.2. Quantification of Apoptosis Using Propidium Iodide and Acridine Orange Double-Staining Apoptotic, necrotic and viable CEMss cells were scored under the fluorescence microscope. These
also included the control cells (untreated); 200 cells were randomly and differentially counted. The
study revealed that TQ triggered morphological features that relate to apoptosis in a time-dependent
manner (Figure 2). Early apoptosis was obvious by intercalated AO within the fragmented DNA. In
several such cases, the fluorescent bright-green colour could only be seen in treated CEMss cells. In
contrast, untreated cells were observed to have a green intact nuclear structure. At 24 h treatment with
TQ, blebbing and nuclear margination were noticed (moderate apoptosis). In addition, late stages of
apoptosis (i.e., presence of orange colour due to the binding of AO to denatured DNA) were observed
after a 48 and 72 h treatment with TQ (Figure 2). Differential scoring of treated CEMss cells (200 cell
population) as exhibited in Figure 3 showed that there was a statistically significant (p < 0.05)
difference in apoptotic positive cells, which indicated clearly that TQ has a time-dependent
apoptogenic effect. 2.1.1. Cell Culture and Viability Assay On the other hand, there was no statistically significant (p > 0.05) difference in
necrotic counts at different times during treatment (24, 48, and 72 h), as displayed in Figure 2. 11222 Molecules 2013, 18 Figure 2. Fluorescent micrographs of acridine orange and propidium iodide double- stained
CEMss cells. Cells were treated at the IC50 of thymoquinone in a time-dependent manner. Cells were cultured in RPMI 1640 media maintained at 37 °C and 5% CO2. (A,B) Untreated cells after 72 h showed normal structure without prominent apoptosis and
necrosis. (C,D) Early apoptosis features were seen after 24 h representing intercalated
acridine orange (bright green) amongst the fragmented DNA, (E, F) Blebbing and orange
color representing the hallmark of late apoptosis were noticed in 48 h treatment,
(G,H) bright red colored secondary necrosis were visible after 72 h. VI: viable cells;
BL: blebbing of the cell membrane; LA: late apoptosis; SN: secondary necrosis. Images
are representative of one of three similar experiments. Molecules 2013, 18 11223 Figure 2. Cont. Figure 2. Cont. Figure 3. Percentages of viable, early apoptotic, late apoptosis and secondary necrotic
cells after thymoquinone treatment. CEMss cell death via apoptosis increased significantly
(* p < 0.05) in a time-dependent manner. However, no significant (p > 0.05) difference
was observed in the cell count of necrosis. VI: viable Cell, EA: Early apoptosis, LA: late
apoptosis, SN: secondary necrosis. Figure 3. Percentages of viable, early apoptotic, late apoptosis and secondary necrotic
cells after thymoquinone treatment. CEMss cell death via apoptosis increased significantly
(* p < 0.05) in a time-dependent manner. However, no significant (p > 0.05) difference
was observed in the cell count of necrosis. VI: viable Cell, EA: Early apoptosis, LA: late
apoptosis, SN: secondary necrosis. 0
10
20
30
40
50
60
70
CONTROL
24H
48H
72H
Percentage of cells (%)
VI
EA
LA
SN
*
*
*
*
*
*
*
*
* Molecules 2013, 18 Molecules 2013, 18 11224 2.1.3. Annexin V The induced apoptotic effect of TQ was further confirmed by the determination of the percentage of
apoptotic cells using flow cytometric analysis with the AV/PI double staining. The AV+/PI_ staining
represents the early apoptotic cells due to the strong affinity of AV-FITC with phosphatidylserine,
which transports from the inner leaflet of the plasma membrane to the outer surface of the membrane
during early apoptosis. On the other hand, AV_/PI+ staining represents the necrotic cells, since PI,
which could not cross through an intact cell membrane, penetrates the compromised membrane of
dead cells or late apoptotic cells and binds to nucleic acid. Meanwhile, viable cells can be marked by
AV_/PI_, and AV+/PI+ staining is indicative of late apoptotic cells. The representative dot plots of the
flow cytometric analysis of apoptosis showed that based on a comparison between untreated cells
(control) and treated cells (24 h and 48 h), the percentages in early apoptosis and late apoptosis
respectively increased (Figure 4). In addition to that, TQ treatment clearly produced a slight decrease
in viable cells at 24 and 48 h. The results further suggest that the antiproliferative effect of TQ against
CEMss cells is caused by inducing cell apoptosis. Figure 4. The effect of thymoquinone on early apoptosis of CEMss cells. CEMss cells
were exposed to1.5 µg/mL and incubated at 37 °C in a CO2 incubator. After staining with
FITC-conjugated Annexin V and PI, cells were analyzed by flow cytometry. Control cells
received no drug treatments. The early apoptotic events (Annexin+/PI-) are shown in lower
right quadrant (Q4-1) of each panel. Quadrant (Q2-1) represents Annexin+/PI+ late stage
of apoptosis/dead cells. (A) The CEMss control (n = 2). (A–C) The effects of 0, 24 and 48 h
exposure (respectively) of CEMss cells to Thymoquinone. 4.1 %
3.4 %
78.8 %
4.7 %
A 4.1 %
3.4 %
78.8 %
4.7 %
A 4.1 %
3.4 %
78.8 %
4.7 %
A 11225 Molecules 2013, 18 Figure 4. Cont. Figure 4. Cont. 2.1.4. DNA Fragmentation Assay
14.9 %
17.9 %
24.6 %
40.9 %
3.2 %
19.7 %
53.3 %
23.8 %
B
C 2.1.4. DNA Fragmentation Assay The DNA ladder pattern of internucleosomal fragmentation (Figure 5), another characteristic of
apoptosis, was distinctly seen in TQ-treated cells. Formation of the DNA ladder pattern also increased
in a time-dependent manner. Molecules 2013, 18 Molecules 2013, 18 11226 2.1.3. Annexin V Positive control in the figure shows the ladder of the DNA whilst the
negative control shows the untreated cells which did not undergo apoptosis. The ladder visibly
increased in a time-dependent manner. Figure 5. Effects of Thymoquinone on DNA fragmentation in CEMss cells. (Lanes A–D
were treated with 1.5 µg/mL. Lane E: positive control and lane F: marker). Figure shows
data representative of three independent experiments. A B
C D E
F 2.1.5. Caspases 3, 8 and 9 Analyses 2.1.5. Caspases 3, 8 and 9 Analyses The apoptotic effect of TQ was also examined by measuring the activities of caspases 3, 8 and 9
enzymes. The enzyme activities were determined in relation to the different concentrations of protein
content (50–250 μg/sample) for cells treated with IC₅₀ of TQ. The results obtained showed an increase
of the enzyme activities in a time-dependent manner [22], as shown in Figure 6. Figure 6. Relative expression of Caspase-3 (A), -8 (B) and -9 (C) in the CEMss cells
treated with different concentrations of Thymoquinone. Triplicates of each treatment group
were used ineach independent experiment. The statistical significance is expressed as *,
p < 0.05. A: Caspase 3, B: Caspase 8, C: Caspase 9. . 0
0.2
0.4
0.6
0.8
1
1.2
1.4
control
24h
48h
72h
Absorbansce 405 nm
Time
*
*
*
A
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
control
24h
48h
72h
Absorbance 405 nm
Time
*
*
*
B . 0
0.2
0.4
0.6
0.8
1
1.2
1.4
control
24h
48h
72h
Absorbansce 405 nm
Time
*
*
*
A 0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
control
24h
48h
72h
Absorbance 405 nm
Time
*
*
*
B 11227 Molecules 2013, 18 Figure 6. Cont. 2.1.6. Cell Cycle Analysis
Both treated and untreated cells were analyzed in terms of cell cycle distribution by means of flow
cytometry. The cell cycle analysis confirmed that TQ induced a depletion of cells in the G1 phase. However, apoptosis increased significantly (Figure 7A) at 3, 6, 12, 24, 48 and 72 h of treatment, which
indicated that these cells had undergone apoptosis (Figure 7). These results suggest that TQ induces S
phase cell cycle arrest followed by apoptosis in CEMss cells [23]. 0
0.5
1
1.5
2
2.5
control
24h
48h
72h
Absorbance 405nm
Time
c
*
*
* 2.1.6. Cell Cycle Analysis
0
0.5
1
1.5
2
2.5
control
24h
48h
72h
Absorbance 405nm
Time
c
*
*
* 2.1.6. Cell Cycle Analysis Both treated and untreated cells were analyzed in terms of cell cycle distribution by means of flow
cytometry. The cell cycle analysis confirmed that TQ induced a depletion of cells in the G1 phase. However, apoptosis increased significantly (Figure 7A) at 3, 6, 12, 24, 48 and 72 h of treatment, which
indicated that these cells had undergone apoptosis (Figure 7). 2.1.5. Caspases 3, 8 and 9 Analyses These results suggest that TQ induces S
phase cell cycle arrest followed by apoptosis in CEMss cells [23]. 2.1.7. Western Blots Bcl-2 and Bax are members of the Bcl-2 family that play key roles in the regulation of apoptosis;
these proteins are believed to be membrane-bound and their ability to undergo both homodimerization
and heterodimerization has been proposed to regulate apoptosis [24]. Although the caspase proteolytic
cascade is a central point in apoptotic response, its activity is tightly regulated by a variety of factors. Among these, Bcl-2 family proteins, including anti-apoptotic members (such as Bcl-2) and
pro-apoptotic members (such as Bax and Hsp70), play a pivotal role. To further analyze the possible
mechanism underlying the TQ-induced apoptosis, we tested the expression of Bcl-2, Bax and Hsp70 in
CEMss cells after the TQ treatment. After being normalized to β-actin, the expression of Bcl-2 and
Hsp70 decreased significantly while Bax protein level increased remarkably in a dose-dependent
manner (Figure 8). The ratio of Bax: Bcl-2 Hsp70 also dramatically increased in a time-dependent
manner [25]. 11228 Molecules 2013, 18 Figure 7. Histograms for cell cycle from analysis of CEMss cells treated with
thymoquinone (1.5 µg/mL) (A). Results are representative of one of three independent
experiments. Induction of S phase arrest in the cell cycle progression of CEMss cells
thymoquinone (B). “*” Indicates a significant difference p < 0.05. Channels (PI-A)
0
20
40
60
80
100
120
Number
0
70
140
210
280
Channels (PI-A)
0
30
60
90
120
Number
0
80
160
240
320
Channels (PI-A)
0
30
60
90
120
Number
0
60
120
180
240
Channels (PI-A)
0
20
40
60
80
100
120
Number
0
40
80
120
160
Control
3 h
6 h
12 h
A Channels (PI-A)
0
20
40
60
80
100
120
Number
0
70
140
210
280
Channels (PI-A)
0
30
60
90
120
Number
0
80
160
240
320
A A Channels (PI-A)
60 Channels (PI A)
Channels (PI A)
Channels (PI-A)
0
30
60
90
120
Number
0
60
120
180
240
Channels (PI-A)
0
20
40
60
80
100
120
Number
0
40
80
120
160
Control
3 h
6 h
12 h 3 h 0
Number
0
60
120
180
240 Number
120
180
240 6 h 12 h 11229 Molecules 2013, 18 Figure 7. Cont. 2.1.7. Western Blots Channels (PI-A)
0
40
80
120
Number
0
10
20
30
40
50
Channels (PI-A)
0
40
80
120
Number
0
10
20
30
40
Channels (PI-A)
0
40
80
120
Number
0
10
20
30
50.00%
60.00%
70.00%
80.00%
90.00%
of cells (%)
G1
G2
S phase
Apoptosis
*
*
*
*
*
*
B
24 h
48 h
72 h Figure 7. Cont. Figure 7. Cont. Channels (PI-A)
0
40
80
120
Number
0
10
20
30
40
50
Channels (PI-A)
0
40
80
120
Number
0
10
20
30
40
24 h
48 h Figure 7. Cont. 0
Number
0
10
20
30
40
50 Channels (PI-A)
0
40
80
120
Number
0
10
20
30
40
50
Channels (PI-A)
0
40
80
120
Number
0
10
20
30
40
Channels (PI-A)
0
40
80
120
Number
0
10
20
30
0.00%
10.00%
20.00%
30.00%
40.00%
50.00%
60.00%
70.00%
80.00%
90.00%
C
3H
6H
12H
24H
48H
72H
Percentage of cells (%)
Time
G1
G2
S phase
Apoptosis
*
*
*
*
*
*
B
24 h
48 h
72 h 24 h Channels (PI-A)
0
40
80
120
Number
0
10
20
30
24 h
48 h
72 h 0.00%
10.00%
20.00%
30.00%
40.00%
50.00%
60.00%
70.00%
80.00%
90.00%
C
3H
6H
12H
24H
48H
72H
Percentage of cells (%)
Time
G1
G2
S phase
Apoptosis
*
*
*
*
*
*
B B B 11230 Molecules 2013, 18 Figure 8. Effect of TQ on the levels of apoptosis regulatory proteins at 3, 6, 12 and 24 h
with β- actin as a loading control. ‘*’ indicates statistically significant at p < 0.05. Figure 8. Effect of TQ on the levels of apoptosis regulatory proteins at 3, 6, 12 and 24 h
with β- actin as a loading control. ‘*’ indicates statistically significant at p < 0.05. A
Bax
Bcl2
HSP70
B-Actin
0 3 6 12 24 h
B
*
*
*
*
*
*
*
*
* A
Bax
Bcl2
HSP70
B-Actin
0 3 6 12 24 h B
*
*
*
*
*
*
*
*
* Bax 2.1.8. Reactive Oxygen Species (ROS) 2.2. Discussion Apoptosis is one form of physiological or active cell death, or a key pathway for regulating
homeostasis and morphogenesis of mammalian cells and is connected with several diseases, especially
cancer [27]. It describes the orchestrated collapse of a cell characterized by membrane blabbing, cell
shrinkage, condensation of chromatin, and fragmentation of DNA followed by a rapid engulfment of
the corpse by neighbouring cells [28]. Apoptosis is becoming widely acknowledged as being an innate
tissue protection against carcinogens by inhibiting survival and controlling the growth of precancerous
cell populations and tumours at different stages of carcinogenesis. Reflecting on this knowledge, the
mechanism of action for many currently used anticancer agents have been specifically targeted to
regulate the apoptotic pathway, further stressing the role of programmed cell death in maintaining
normal homeostasis. A major concern of cancer chemotherapy is the side effects caused by the
non-specific targeting of both normal and cancerous cells by therapeutic drugs. Much emphasis has
been placed on discovering new compounds that target tumour cells more efficiently and selectively
with minimal toxic effects on normal cells [29]. Therefore, several researchers nowadays have
performed anti-cancer studies on herbal extracts as well as natural compounds based on their
biochemical properties of apoptosis [30,31]. Thymoquinone is the major bioactive constituent present in black seed oil (Nigella sativa) and has
been tested for its efficacy against cancer [16,32,33]. Many researchers have studied TQ’s mechanism
of action and its aptitude to induce apoptosis and inhibit tumour growth [34]. To date, the
chemotherapeutic potential of TQ in the clinic has not been tested [35]. However, numerous studies
have shown its promising anti-cancer effects in animal models [36]. In the present research we have
shown that the cyctotoxic activities of TQ towards CEMss cells are selective (Figure 1). Since the
cytotoxicity was found to be selective, we extended the study to determine the mechanism of
cytotoxicity. Acridine orange (AO) and propidium iodide (PI) are among the most used fluorescent
dyes to analyse cell culture viability and morphological charecteretics. We have observed various
stages of apoptosis, starting from chromatin condensation till apoptotic body formation with TQ
treatment. Even though the morphological features were clearly observed, in an attempt to quantify the
cells of apoptotic population the annexin V assay was performed. Molecules 2013, 18 Molecules 2013, 18 Molecules 2013, 18 11231 2.1.8. Reactive Oxygen Species (ROS) Reactive oxygen species (ROS) are a variety of molecules and free radicals derived from molecular
oxygen which are constantly generated and eliminated in the biological system and have important
roles in cell signalling and homeostasis. Excessive amounts of ROS can cause oxidative damage to
lipids, proteins and DNA, leading to tumour genesis or cell death. Although the use of antioxidants in
humans for cancer prevention remains controversial, increasing evidence have supported that the
increase of ROS generation contributes to the treatment of cancer cells. As shown in Figure 9, we
found that TQ exerted a facilitator role in the production of ROS in a concentration-dependent manner,
which increased the ROS level on CEMss cell lines [26]. Figure 9. Effect of thymoquinone treatment on ROS generation. ROS concentration was
determined by treating CEMss cells with 0, 5, 10 and 20 μg/mL TQ, the cells were loaded
with DCFH-DA, and the fluorescence was measured by a fluorescence micro plate reader. 0
1000
2000
3000
4000
5000
6000
7000
8000
9000
0 μg/mL
5 μg/mL
10 μg/mL
20 μg/mL
Fluorescence 485 nm- 520 nm
Thymoquinone Molecules 2013, 18 Caspase-3 is required for typical hallmarks of apoptosis, and is indispensable for apoptotic chromatin
condensation and DNA fragmentation in all cell types examined. Thus, caspase-3 is essential for
certain processes associated with the dismantling of the cell and the formation of apoptotic bodies, but
it may also function before or at the stage when commitment to loss of cell viability is made [43]. Caspase-8 is a family member of executioner caspases associated with tumour necrosis factor (TNF)
family death receptors-mediated apoptotic signalling cascade [44]. In this study, the results of caspases
3, 8 and 9 activities significantly showed that TQ induces apoptosis in CEMss in a time-dependent
manner which is caspase dependent. Phytochemicals that function as cell-cycle modulators are gaining widespread attention due to the
evidence of concomitant involvement of apoptosis and cell cycle inhibition. The cell cycle has four
sequential phases. Arguably, the most important phases are the S phase, when DNA replication occurs,
and the M phase, when the cell divides into two daughter cells. During the past two decades, cancer
genetics have shown that hyperactivating mutations in growth signalling networks, coupled with loss
of function of tumour suppressor proteins, drive oncogenic proliferation. The cell cycle machinery,
which acts as an integration point for information transduced through upstream signalling networks,
represents an alternative target for diagnostic and therapeutic interventions. Analysis of the DNA
replication initiation machinery and mitotic engine proteins in human tissues is now leading to the
identification of novel biomarkers for cancer detection and prognostication, and is providing target
validation for cell cycle-directed therapies [45]. Thus flow cytometry analysis was performed to
analyze the various cell cycle check points at TQ treatment. Our results indicate that cell cycle
distribution is altered in CEMss cells after TQ treatment. This growth inhibition was mediated through
blocking progression of the leukemia cells through the S phase of the cell cycle, as previously
described [46]. Reactive oxygen species (ROS) are constantly generated and eliminated in the biological system,
and play important roles in a variety of normal biochemical functions and abnormal pathological
processes. Consequently, humans have evolved antioxidant defence systems that limit their production. Generation of oxidative stress in response to various external stimuli has been implicated in the
activation of transcription factors and with the triggering of apoptosis [47]. 2.2. Discussion Annexin V was shown to interact
strongly and specifically with PS and could be used to detect apoptosis by targeting the loss of plasma
membrane asymmetry [37]. The present study exhibits that treatment with TQ is able to induce cell death
via apoptosis in human CEMss cells. The results showed that there was a significant time-dependent
increase in the early stage of apoptosis, as shown clearly in Figure 4. In addition to annexin V analysis,
the late stage of apoptosis has been confirmed by using a DNA laddering assay, which will identify the
cleavage of chromosomal DNA into oligonucleosomal-sized fragments is an integral part of apoptosis [38]. Moreover this assay was usded before many time to detect the apoptosis in leukemia cell lines[39,40]. Apoptosis is orchestrated by a family of cysteine proteases known as caspases. The main effectors
of apoptosis encompass proteases from the caspase family, which reside as latent precursors in most
nucleated animal cells [41]. Fourteen mammalian caspases have been identified, three of which
(caspase-3, -6, and -7) are thought to coordinate the execution phase of apoptosis by cleaving multiple
structural and repair proteins [42]. Pathways to caspase-3 activation have been identified, which are
either dependent on or independent of mitochondrial cytochrome c release and caspase-9 function. 11232 Molecules 2013, 18 Growing evidence suggests
that cancer cells exhibit increased intrinsic ROS stress, due in part to oncogenic stimulation, increased
metabolic activity and mitochondrial malfunction. Previously TQ-induced apoptosis of chondrocytes
has been reported to be involved in the generation of ROS [48]. In agreement with this we found that
TQ is a potent inducer of apoptosis in CEMss cells via release of ROS. The generation of ROS is
cloely associated with mitochondrial proteins such as Bcl-2 and Bax. These proteins are believed to be
membrane-bound and their ability to undergo both homodimerization and heterodimerization has been
proposed to regulate apoptosis [49]. The Bcl-2 family proteins are key regulators of apoptosis, which
include both anti- and pro-apoptotic proteins and a slight change in the dynamic balance of these
proteins, may result in either inhibition or promotion of cell death [50]. Meanwhile, there are many
prtoiens inside the cells which will prevent the ROS mediated cell dammage. Amongst them, HSP70 is
widely studies for its protection fuction. It is a chaperone that accumulates in the cells after many
different stresses promoting cell survival in response to the adverse conditions. In contrast to normal
cells, most cancer cells richly express HSP70 at the basal level to resist various insults at different stages 11233 Molecules 2013, 18 of tumourigenesis and during anti-cancer treatment [51]. We have found that TQ caused a significant
down-regulation of Bcl2 and HSP70 and an up-regulation of BAX in a time-dependent manner. 3.1. Chemicals and Reagents Thymoquinone (>99% pure) was purchased from Sigma (St. Louis, MO, USA). CEMss cell lines
used in this study were obtained from the NIH AIDS Research and Reference Reagent Program,
Division of AIDS, NIAID, NIH: Bethesda, MD, USA. RPMI 1460, fetal bovine serum (FBS) and
penicillin-streptomycin were obtained from Bioscience Ltd. (Wokingham, UK). Phosphate buffered
saline (PBS), ethanol (95%), DNA laddering kit, 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium
bromide (MTT) and DCFH-DA (2'-7'-dichlorodihydrofluorescein diacetate were from Santa Cruz
Biotechnology (Santa Cruz, CA USA), propidium iodide and acridine orange was purcahed from
Nacalai Tesque (Japan), Annexin V kit was from ClonTech (Palo Alto, CA, USA). 3.2. Cell Culture and Viability Assay Cell cultures were maintained in a humidified atmosphere with 5% CO2 at 37 °C. To study
proliferation and determine the survival of cancer cells after treatment, the MTT assay was used. Briefly, the cells were plated on a 96-well plate at 2 × 105 cell/mL in 100 μL culture medium. They
were plated in triplicate. Different concentrations of TQ (50, 25, 12.5, 6, 3 and 1.5 µg/mL) were
prepared by serial dilution. All serial dilutions were transferred to the cells in the 96-well plate; the
plate included untreated cells as control and was incubated for 24 h. After incubation, the viability of
the cells was assessed using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT,
5 mg/mL); 20 μL were added to the cells in a dark place, and the cells were then covered with
aluminium foil and incubated for 4 h. After incubation, all of the media were removed and 100 μL of
DMSO were added to the cells in order to solubilise the formazan crystals. Subsequently, the
absorbance was read at a wavelength of 570 nm using a micro plate reader. The potency of cell growth
inhibition for the test agent was expressed as an IC50 value. 3.3. Quantification of Apoptosis Using Propidium Iodide and Acridine Orange Double-Staining TQ-induced cell death in CEMss leukemia cells was quantified using propidium iodide (PI) and
acridine orange (AO) double-staining according to standard procedures and examined under a
fluorescence microscope (Lieca attached with Q-Floro Software, Solms, Germany). Briefly, treatment
was carried out in a 25-mL culture flask (Nunc, Roskilde, Denmark). CEMss cells were plated at a
concentration of 2 × 105 cell/mL and treated with TQ at the IC50 concentration. Flasks were incubated
in an atmosphere of 5% CO2 at 37 °C for 24 and 48 h. The cells were then spun down at 1,800 rpm for
10-min. Supernatant was discarded and the cells were washed twice using cold phosphate buffered
saline (PBS) after being centrifuged at 1,800 rpm for 10-min to remove the remaining media. Five
microliters of fluorescent dye containing AO (10 μg/mL) and PI (10 μg/mL) were added into the
cellular pellet at equal volumes. Freshly stained cell suspension was dropped into a glass slide and
covered by a cover slip. Slides were then observed under the fluorescence microscope within 30-min 11234 Molecules 2013, 18 before the fluorescent colour started to fade. The percentages of viable, early apoptotic, late apoptosis
and secondary necrotic cells were determined in >200 cells. AO and PI are intercalating nucleic
acid-specific fluorochromes which emit green and orange fluorescence, respectively, when they are
bound to DNA. Of the two, only AO is a membrane-permeable, cationic dye that binds to nucleic acids
of viable cells and at low concentrations causes a green fluorescence. PI is impermeable to intact
membranes but readily penetrates the membranes of nonviable cells and binds to DNA or RNA,
resulting in orange fluorescence. When AO and PI are used simultaneously, viable cells fluoresce
green and nonviable cells fluoresce orange under the fluorescence microscopy. The criteria for
identification are as follows: (i) viable cells appear to have green nucleus with intact structure;(ii) early
apoptosis exhibits a bright-green nucleus showing condensation of chromatin in the nucleus; (iii) dense
orange areas of chromatin condensation showing late apoptosis; and (iv) orange intact nucleus
depicting secondary necrosis [52]. 3.4. Annexin V Assay CEMss cells (2 × 105 cells/mL) were exposed to the IC50 concentration for 24 and 48 h and the
Annexin V assay was performed using the BD Pharmingen TM Annexin V-FITC Apoptosis Detection
Kit (APO Alert Annexin V, ClonTech). Briefly, treated cells were centrifuged for 10 min at 200 × g to
remove the media. Later, the cells were rinsed with 1 × binding buffer supplied by the manufacturer. The rinsed cells were resuspended in binding buffer (200 μL) and annexin V (5 μL) and propidium
iodide (10 μL) were added and the cells were then incubated at room temperature in the dark for
15 min. Flow cytometric analysis was carried out using a BD FACSCanto™ II instrument The binding
buffer supplied by the manufacturer was used to bring the reaction volume to at least 500 μL for the
flow cytometry analysis. DMSO-treated (0.1%, v/v) CEMss cells were used as control. 3.6. Caspase 3, 8 and 9 Activities In vitro determination of the proteolytic activity of the enzymes in lysates of CEMss cells was
performed. Induced apoptosis in cells by TQ at different times (12, 24 and 48 h) was detected using the
R&D Systems Kit. Briefly, cells were collected, washed with cold PBS (1,800 rpm in 10 min) and
subsequently resuspended in protein lysis buffer and incubated for 10 min on ice. Then, centrifugation
was done at 10,000 g for 1 min, and the supernatant (protein) was collected. Fifty microliters of protein
were transferred to 96-well plates in triplicates, then 50 μL of reaction buffer and 5 μL of caspase were
added to each well and they were then incubated for 1–2 h at 37 °C. The plates were read using the
microplate reader at a wavelength of 405 nm [45]. 3.8. Protein Detection by Western Blotting 3.8. Protein Detection by Western Blotting Molecules 2013, 18 Molecules 2013, 18 11235 3.6. Caspase 3, 8 and 9 Activities 3.7. Cell Cycle Analysis CEMss cells at a concentration of 2 × 105 cell/mL were cultured in RPMI 1640 medium containing
10% FBS and 1% penicillin/streptomycin seeded into a 25-mL culture flask (TPP Brand) and treated
with TQ at IC50 concentration for 3, 6, 12, 24, 48 and 72 h. Following incubation, the cells were spun
down at 1,800 rpm for 5 min. The supernatant was discarded and the pellet was washed with PBS
(phosphate buffer saline) twice to remove any remaining media. To restore the integrity, a fixation of
cell population for flow cytometry analysis was performed. Briefly, cell pellets were fixed by mixing
700 μL of 90% cold ethanol and keeping them at 4 °C overnight. The cells were then spun down at 200 g
for 5 min and the ethanol was decanted. After being washed once with PBS, cells were resuspended in
PBS (600 μL) and RNase (25 μL, 10 mg/mL) and propidium iodide (PI, 50 μL, 1 mg/mL) were added
to the fixed cells and they were kept for 1 h at 37 °C. PI has the ability to bind to RNA molecules and
hence, RNase enzyme was added in order to allow PI to bind directly to the DNA. The DNA content
of the cells was then analyzed using the flow cytometer. The fluorescence intensity of sub-G1 cell
fraction represented the apoptotic cell population. 3.5. DNA Fragmentation Assay Apoptosis was confirmed through detection of the fragmentation of chromosomal DNA with the
DNA ladder method. The Apoptotic DNA Ladder Detection Kit (Chemicon International Inc., Palo
Alto, CA, USA) was used to extract DNA from the cells. The cells were treated time dependently
using IC50 concentration for 24, 12 and 6 h. Briefly, 2 × 105 cell/mL were washed twice using cool
phosphate buffered saline (PBS) and the cells were collected by centrifuging them at 300 × g for
10-min. Following removal of the supernatant, the cells were lysed by adding 40 μL of TE (Tris and
EDTA) lysis buffer, followed by 5 μL of Enzyme A (RNase A) and incubated at 37 °C for 10 min. Five microliters of Enzyme B (Proteinase K) were added and the lysate was further incubated at 50 °C
for 30 min. Consequently, five microliters of ammonium acetate solution and 50 μL of isopropanol
were added to the lysate, mixed well and kept at −20 °C for 10 min. The samples were then
centrifuged for 10 min at 16,000 × g to precipitate the DNA. After washing, 70% ice cold ethanol was
added to the DNA pellet. The pellet was then air dried and later dissolved in 30 μL of DNA suspension
buffer. The extracted DNA samples were run on a 1.5% agarose gel in Tris-acetic acid-EDTA buffer. After electrophoresis, the gel was stained with ethidium bromide (Gibco BRL, Dakkopatts, Scotland)
and the band obtained was visualized using a UV light transilluminator. 4. Conclusions The anticancer activity of TQ in acute lymphoblastic leukaemia was studied by various methods
including cell viability assay, AO/PI, DNA laddering, flow-cytometric analysis, caspase-3 activity and
western blotting analysis. Results from all the experiments suggested that TQ could significantly
inhibit proliferation and induce apoptosis in the leukemic cell line CEMss. 3.10. Statistical Analysis Results were reported as means ± SEM of at least three analyses for each sample. Normality and
homogeneity of variance assumptions were checked. Statistical analysis was performed according to
the SPSS-16.0 package. Acknowledgements The authors would like to express their utmost gratitude to the late A Hamid Bin A. Hadi for his
invaluable support. Gratitude also extends to University of Malaya (PG141-2012B) and Ministry of
Higher Education (HIR grant F00009-21001) for providing the grant to conduct this study. 3.8.2. Western Blotting Analysis Forty micrograms of protein extract was separated by 10% SDS-PAGE, transferred to a
polyvinylidenedifluoride (PVDF) membrane (Bio-Rad, Hercules, CA, USA) and blocked with 5%
non-fat milk in TBS-Tween buffer 7 (0.12 M Tris-base, 1.5 M NaCl, 0.1% Tween20) for 1 h at room
temperature. Afterwards, it was incubated overnight with the appropriate antibody at 4 °C and then
incubated with horseradish peroxidase-conjugated secondary antibody for 30 min at room temperature. The bound antibody was detected using peroxidase-conjugated anti-rabbit antibody (1:10000) or
anti-mouse antibody (1:10000) followed by chemiluminescence and exposed by autoradiography. The
primary antibodies β-actin (1:10,000), Bcl2 (1:1000), Bax (1:1000) and HSP70 (1:1000) were
purchased from Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA. 3.8.1. Extraction of Whole Protein from the Cells CEMss cells at a concentration of 2 × 105 cells/mL were cultured in RPMI 1640 (PAA, Coelbe,
Germany) medium containing 10% FBS, seeded into a 75-mm culture flask (TPP Brand) and then
treated with IC50 concentration for 3, 6, 12 and 24 h. After incubation, the cells were spun down at
1,000 rpm for 10 min. The supernatant was discarded and the pellet was washed twice with phosphate
buffered saline (PBS) to remove any remaining media. Estimation of the packed cell pellet volume was
done and 20 volumes of mammalian cell lysis reagent (Proteo JET, Fermentas Life Sciences, Burlington,
ON, Canada) were added to 1 volume of packed cells. The cells were then incubated for 10 min at room
temperature on a shaker (900–1,200 rpm) and centrifugation was done at 16,000–20,000 × g for 15 min
to clarify the lysate. The resultant lysate was then transferred to a new tube and stored at −70 °C until
analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Molecules 2013, 18 Molecules 2013, 18 11236 3.9. Determination of Reactive Oxygen Species (ROS) The production of intracellular ROS was measured using 2', 7'-dichlorofluorescin diacetate
(DCFH-DA). Briefly, 10 mM DCFH-DA stock solution (in methanol) was diluted 500-fold in Hank's
Balanced Salt Solution (HBSS) without serum or other additives to yield a 20 μM working solution. After 24 h of exposure to TQ, the cells in the 96-well black plate were washed twice with HBSS and
then incubated in 100 μL working solution of DCFH-DA at 37 °C for 30 min. Fluorescence was then
determined at 485-nm excitation and 520-nm emission using a fluorescence microplate reader
(Tecan Infinite M 200 PRO, Männedorf, Switzerland) [43]. References 1. Yaman, İ.; Balikci, E. Protective effects of Nigella sativa against gentamicin-induced
nephrotoxicity in rats. Exp. Toxicol. Pathol. 2010, 62, 183–190. 1. Yaman, İ.; Balikci, E. Protective effects of Nigella sativa against gentamicin-induced
nephrotoxicity in rats. Exp. Toxicol. Pathol. 2010, 62, 183–190. 2. Hawsawi, Z.A.; Ali, B.A.; Bamosa, A.O. Effect of Nigella sativa (black seed) and thymoquinone
on blood glucose in albino rats. Ann. Saudi. Med. 2001, 21, 242–244. 2. Hawsawi, Z.A.; Ali, B.A.; Bamosa, A.O. Effect of Nigella sativa (black seed) and thymoquinone
on blood glucose in albino rats. Ann. Saudi. Med. 2001, 21, 242–244. 3. Ahmed, W.A.; Hassan, S.A.; Galeb, F.M.; El-Taweel, M.A.; Abu-Bedair, F.A. The in vitro
promising therapeutic activity of thymoquinone on hepatocellular carcinoma (HepG2) cell line. Global Vet. 2008, 2, 233–241. 3. Ahmed, W.A.; Hassan, S.A.; Galeb, F.M.; El-Taweel, M.A.; Abu-Bedair, F.A. The in vitro
promising therapeutic activity of thymoquinone on hepatocellular carcinoma (HepG2) cell line. Global Vet. 2008, 2, 233–241. 4. Abu-Irmaileh, B. E.; Afifi, F. U. Herbal medicine in Jordan with special emphasis on commonly
used herbs. J. Ethnopharmacol. 2003, 89, 193–197. 4. Abu-Irmaileh, B. E.; Afifi, F. U. Herbal medicine in Jordan with special emphasis on commonly
used herbs. J. Ethnopharmacol. 2003, 89, 193–197. 5. Ramadan, M.F. Nutritional value, Functional properties and nutraceutical applications of
black cumin (Nigella sativa L.): An Overview. Int. J. Food Sci. Technol. 2007, 42,
1208–1218. 6. Said, O.; Khalil, K.; Fulder, S.; Azaizeh, H. Ethnopharmacological survey of medicinal herbs in
Israel, the Golan Heights and the West Bank region. J. Ethnopharmacol. 2002, 83, 251–265. 7. Shah, S.; Kasturi, S.R. Study on antioxidant and antimicrobial properties of black cumin
(Nigella sativa Linn). J. Food Sci. Technol. 2003, 40, 70–73. 8. Ghannadi, A.; Hajhashemi, V.; Jafarabadi, H. An investigation of the analgesic and
anti-inflammatory effects of Nigella sativa seed polyphenols. J. Med. Food. 2005, 8, 488–493. 9. Singh, G.; Marimuthu, P.; de Heluani, C.S.; Catalan, C. Chemical constituents and antimicrobial
and antioxidant potentials of essential oil and acetone extract of Nigella sativa seeds. J. Sci. Food Agric. 2005, 85, 2297–2306. 10. Sayed-Ahmed, M.M.; Aleisa, A.M.; Al-Rejaie, S.S.; Al-Yahya, A.A.; Al-Shabanah, O.A.;
Hafez, M.M.; Nagi, M.N. Thymoquinone attenuates diethylnitrosamine induction of
hepatic carcinogenesis through antioxidant signaling. Oxid. Med. Cell. Longev. 2010, 3,
254–261. 11. Nasaruddin, N. Extraction of pharmacologically active thymoquinone in Nigella sativa L. Master’s Thesis. Universiti Malaysia Pahang: Pahang, Malaysia, 2006. 12. Arafa, E.-S. Molecules 2013, 18 Molecules 2013, 18 11237 Conflicts of Interest The authors declare no conflict of interest. Molecules 2013, 18 References A.; Zhu, Q.; Shah, Z. I.; Wani, G.; Barakat, B. M.; Racoma, I.; El-Mahdy, M. A.;
Wani, A.A. Thymoquinone up-regulates PTEN expression and induces apoptosis in
doxorubicin-resistant human breast cancer cells. Mutat. Res. 2011, 706, 128–135. 13. Banerjee, S.; Azmi, A.S.; Padhye, S.; Singh, M.W.; Baruah, J.B.; Philip, P.A.; Sarkar, F.H.;
Mohammad, R.M. Structure-activity studies on therapeutic potential of Thymoquinone analogs in
pancreatic cancer. Pharmaceut. Res. 2010, 27, 1146–1158. 14. Jafri, S.H.; Glass, J.; Shi, R.; Zhang, S.; Prince, M.; Kleiner-Hancock, H. Thymoquinone and
cisplatin as a therapeutic combination in lung cancer: in vitro and in vivo. J. Exp. Clin. Cancer Res. 2010, 29, 2285. 15. Woo, C.C.; Loo, S.Y.; Gee, V.; Yap, C.W.; Sethi, G.; Kumar, A.P.; Benny Tan, K.H. Anticancer
activity of thymoquinone in breast cancer cells: possible involvement of PPAR-γ pathway. Biochem. Pharmacol. 2011, 82, 464–475. Molecules 2013, 18 11238 16. Gali-Muhtasib, H.; Kuester, D.; Mawrin, C.; Bajbouj, K.; Diestel, A.; Ocker, M.; Habold, C.;
Foltzer-Jourdainne, C.; Schoenfeld, P.; Peters, B. Thymoquinone triggers inactivation of the stress
response pathway sensor CHEK1 and contributes to apoptosis in colorectal cancer cells. Cancer Res. 2008, 68, 5609–5618. 17. Amin, A.; Gali-Muhtasib, H.; Ocker, M.; Schneider-Stock, R. Overview of major classes of
plant-derived anticancer drugs. Int. J. Biomed. Sci. 2009, 5, 1–11. 18. Lim, G.C.C. Overview of cancer in Malaysia. Jpn. J. Clin. Oncol. 2002, 32, S37–S42. 19. Trigg, M.E.; Sather, H.N.; Reaman, G.H.; Tubergen, D.G.; Steinherz, P.G.; Gaynon, P.S.;
Uckun, F.M.; Hammond, G.D. Ten-year survival of children with acute lymphoblastic leukemia:
A Report from the Children’s Oncology Group. Leuk. Lymphoma. 2008, 49, 1142–1154. 20. Wagiman, S.N.; Husin, N.H.; Latiff, Z.A.; Alias, H.; Jamal, R.; Zakaria, S.Z.S. Folate-related
genes polymorphisms associated with the risk of childhood leukaemia in the Malaysian
population. Asia. Pac. J. Mole. Med. 2011, 1, 1–10. 21. Zhu, J. Y.; Lavrik, I. N.; Mahlknecht, U.; Giaisi, M.; Proksch, P.; Krammer, P. H.; Li-Weber, M. The traditional Chinese herbal compound rocaglamide preferentially induces apoptosis in
leukemia cells by modulation of mitogen-activated protein kinase activities. Int. J. Cancer 2007,
121, 1839–1846. 22. Sosne, G.; Siddiqi, A.; Kurpakus-Wheater, M. Thymosin-β4 inhibits corneal epithelial cell
apoptosis after ethanol exposure in vitro. Invest. Ophthalmol. Visual. Sci. 2004, 45, 1095–1100. 23. Chen, H.W.; Huang, H.C. Effect of curcumin on cell cycle progression and apoptosis in vascular
smooth muscle cells. Brit. J. Pharmacol. 1998, 124, 1029–1040. 24. Hsu, Y.-T.; Wolter, K.G.; Youle, R.J. References Cytosol-to-membrane redistribution of Bax and Bcl-XL
during apoptosis. Proc. Nat. Acad. Sci. Usa 1997, 94, 3668–3672. 25. Ye, M.; Liu, J.-K.; Lu, Z.-X.; Zhao, Y.; Liu, S.-F.; Li, L.-L.; Tan, M.; Weng, X.-X.; Li, W.; Cao, Y. Grifolin, a potential antitumor natural product from the mushroom Albatrellus confluens, inhibits
tumor cell growth by inducing apoptosis in vitro. Febs. Lett. 2005, 579, 3437–3443. 26. Jia, N.; Xiong, Y.L.; Kong, B.; Liu, Q.; Xia, X. Radical scavenging activity of black currant
(Ribes nigrum L.) extract and its inhibitory effect on gastric cancer cell proliferation via induction
of apoptosis. J. Funct. Foods 2012, 4, 382–390. 27. Goldsworthy, T.L.; Conolly, R.B.; Fransson-Steen, R. Apoptosis and cancer risk assessment. Mutat. Res-Rev. Genet. Toxicol. 1996, 365, 71–90. 28. Taraphdar, A.K.; Roy, M.; Bhattacharya, R. Natural products as inducers of apoptosis:
Implication for cancer therapy and prevention. Curr. Sci. 2001, 80, 1387–1396. 29. Edris, A.E. Anti-cancer properties of nigella spp. essential oils and their major constituents,
Thymoquinone and-Elemene. Curr. Clin. Pharmacol. 2009, 4, 43–46. 30. Kaseb, A.O.; Chinnakannu, K.; Chen, D.; Sivanandam, A.; Tejwani, S.; Menon, M.; Dou, Q.P.;
Reddy, G.P.-V. Androgen Receptor–and E2F-1–Targeted Thymoquinone Therapy for Hormone-
Refractory Prostate Cancer. Cancer Res. 2007, 67, 7782–7788. 31. Banerjee, S.; Padhye, S.; Azmi, A.; Wang, Z.; Philip, P.A.; Kucuk, O.; Sarkar, F.H.; Mohammad,
R.M. Review on molecular and therapeutic potential of thymoquinone in cancer. Nutr. Cancer
2010, 62, 938–946. Molecules 2013, 18 11239 32. Yi, T.; Cho, S.-G.; Yi, Z.; Pang, X.; Rodriguez, M.; Wang, Y.; Sethi, G.; Aggarwal, B.B.; Liu, M. Thymoquinone inhibits tumor angiogenesis and tumor growth through suppressing AKT and
extracellular signal-regulated kinase signaling pathways. Mol. Cancer Ther. 2008, 7, 1789–1796. 32. Yi, T.; Cho, S.-G.; Yi, Z.; Pang, X.; Rodriguez, M.; Wang, Y.; Sethi, G.; Aggarwal, B.B.; Liu, M. Thymoquinone inhibits tumor angiogenesis and tumor growth through suppressing AKT and
extracellular signal-regulated kinase signaling pathways. Mol. Cancer Ther. 2008, 7, 1789–1796. 33. Al-Johar, D.; Shinwari, N.; Arif, J.; Al-Sanea, N.; Jabbar, A.A.; El-Sayed, R.A.; Mashhour, A.;
Billedo, G.; El-Doush, I.; Al-Saleh, I. Role of Nigella sativa and a number of its antioxidant
constituents towards azoxymethane-induced genotoxic effects and colon cancer in rats. Phytother. Res. 2008, 22, 1311–1323. 34. Ivankovic, S.; Stojkovic, R.; Jukic, M.; Milos, M.; Milos, M.; Jurin, M. The antitumor activity of
thymoquinone and thymohydroquinone in vitro and in vivo. Exp. Oncol. 2006, 28, 220–224. 35. Rooney, S.; Ryan, M. References Modes of action of alpha-hederin and thymoquinone, active constituents of
Nigella sativa, against HEp-2 cancer cells. Anticancer Res. 2005, 25, 4255–4259. 36. Van Engeland, M.; Nieland, L.J.; Ramaekers, F.C.; Schutte, B.; Reutelingsperger, C.P. Annexin
V-affinity assay: A Review on an apoptosis detection system based on phosphatidylserine
exposure. Cytometry 1998, 31, 1–9. 37. Nagata, S. Apoptotic DNA fragmentation. Exp. Cell. Res. 2000, 256, 12–18. 38. Faheina-Martins, G.V.; Silveira, A.L.D.; Cavalcanti, B.C.; Ramos, M.V.; Moraes, M.O.; Pessoa, C.;
M Araújo, D.A. Antiproliferative effects of lectins from Canavalia ensiformis and Canavalia
brasiliensis in human leukemia cell lines. Toxicol. Vitro. 2012, 26, 1161–1169. 39. Sari, N.K. DNA Fragmentation on HCT-116 Colon Cancer Cell in the Presence of Short Chain
Fatty Acid (SCFA) Extract. Available online: http://repository.ipb.ac.id/handle/123456789/55761
(accessed on 18 January 2013). 40. Kizhakkayil, J.; Thayyullathil, F.; Chathoth, S.; Hago, A.; Patel, M.; Galadari, S. Modulation of
curcumin-induced Akt phosphorylation and apoptosis by PI3K inhibitor in MCF-7 cells. Biochem. Biophys. Res. Commun. 2010, 394, 476–481. 41. Holmgren, A. Biochemistry: SNO removal. Science 2008, 320, 1019–1020. 42. Williams, G.H.; Stoeber, K. The cell cycle and cancer. J. Pathol. 2012, 226, 352–36 43. Hsuuw, Y.D.; Chang, C.K.; Chan, W.H.; Yu, J.S. Curcumin prevents methylglyoxal-induced
oxidative stress and apoptosis in mouse embryonic stem cells and blastocysts. J. Cell. Physiol. 2005, 205, 379–386. 44. Kruidering, M.; Evan, G.I. Caspase–8 in Apoptosis: The Beginning of “The End”? Iubmb Life
2000, 50, 85–90. 45. Lin, S.-S.; Huang, H.-P.; Yang, J.-S.; Wu, J.-Y.; Hsai, T.-C.; Lin, C.-C.; Lin, C.-W.; Kuo, C.-L.;
Gibson Wood, W.; Chung, J.-G. DNA damage and endoplasmic reticulum stress mediated
curcumin-induced cell cycle arrest and apoptosis in human lung carcinoma A-549 cells through
the activation caspases cascade-and mitochondrial-dependent pathway. Cancer Lett. 2008, 272,
77–90. 46. Motaghed, M.; Al-Hassan, F.; Hamid, S. Cellular responses with thymoquinone treatment in
human breast cancer cell line MCF-7. Pharmacog. Res. 2013, 5, 200–206. 47. Goloudina, A.R.; Demidov, O.N.; Garrido, C. Inhibition of HSP70: A challenging anti-cancer
strategy. Cancer Lett. 2012, 325, 117–124 48. Yu, S.-M.; Kim, S.-J. Thymoquinone-induced reactive oxygen species causes apoptosis of
chondrocytes via PI3K/Akt and p38kinase pathway. Exp. Biol. Med. 2013, in press. Molecules 2013, 18 11240 49. Salvesen, G.S.; Riedl, S.J. Caspase mechanisms. In Programmed cell death in cancer progression
and therapy, Springer: La Jolla, CA, USA, 2007; pp. 13–23. 49. Salvesen, G.S.; Riedl, S.J. Caspase mechanisms. © 2013 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article
distributed under the terms and conditions of the Creative Commons Attribution license
(http://creativecommons.org/licenses/by/3.0/). References In Programmed cell death in cancer progression
and therapy, Springer: La Jolla, CA, USA, 2007; pp. 13–23. 50. Slee, E.A.; Adrain, C.; Martin, S.J. Executioner caspase-3,-6, and-7 perform distinct, non-
redundant roles during the demolition phase of apoptosis. J. Biol. Chem. 2001, 276, 7320–7326. 51. Porter, A.G.; Jänicke, R.U. Emerging roles of caspase-3 in apoptosis. Cell Death Differentiation. 1999, 6, 99–104. 52. Mohan, S.; Abdul, A.B.; Abdelwahab, S.I.; Al-Zubairi, A.S.; Sukari, M.A.; Abdullah, R.;
Elhassan Taha, M.M.; Ibrahim, M.Y.; Syam, S. Typhonium flagelliforme induces apoptosis in
CEMss cells via activation of caspase-9, PARP cleavage and cytochrome c release: Its activation
coupled with G0/G1 phase cell cycle arrest. J. Ethnopharmacol. 2010, 131, 592–600. Sample Availability: Samples of the compounds Thymoquinone are available from the authors. © 2013 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article
distributed under the terms and conditions of the Creative Commons Attribution license
(http://creativecommons.org/licenses/by/3.0/). © 2013 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article
distributed under the terms and conditions of the Creative Commons Attribution license
(http://creativecommons.org/licenses/by/3.0/). © 2013 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article
distributed under the terms and conditions of the Creative Commons Attribution license
(http://creativecommons.org/licenses/by/3.0/).
|
https://openalex.org/W1930023128
|
http://periodicos.ufc.br/rene/article/download/4755/3524
|
Portuguese
| null |
Qualidade na atenção hospitalar ao recém-nascido
|
Rev Rene
| 2,009
|
cc-by
| 4,871
|
QUALIDADE NA ATENÇÃO HOSPITALAR AO RECÉM-NASCIDO
QUALITY IN THE HOSPITAL ATTENTION TO THE NEWBORN
CALIDAD EN LA ATENCIÓN DEL HOSPITAL AL RECIÉN NACIDO MICHELLY KIM DE OLIVEIRA ROSA1
MARIA APARECIDA MUNHOZ GAÍVA2 A qualidade abrange diversos setores e áreas na saúde. Por ser um tema complexo, a avaliação da qualidade tem diferentes
dimensões, sendo as mais utilizadas pelos serviços de saúde, as que envolvem estrutura, processo e resultado. Objetivou-se
refl etir sobre a qualidade e o uso de seus critérios avaliativos no âmbito da assistência neonatal. Trata-se de uma revisão
de literatura, em uma perspectiva teórico-refl exiva. A qualidade da atenção neonatal é resultante da história acerca da as-
sistência ao recém-nascido, da estrutura dos serviços, dos recursos humanos, dos materiais e procedimentos tecnológicos. Existem diferentes maneiras de se avaliar a qualidade da atenção neonatal, incluindo o uso do indicador de mortalidade e
indicadores específi cos da assistência. As discussões oferecidas mostraram a importância e a necessidade de implementação
de práticas avaliativas, na busca por serviços/assistência de qualidade em neonatologia. PALAVRAS-CHAVE: Qualidade da assistência à saúde; Avaliação em saúde; Assistência à saúde; Recém-nascido. Quality comprises several areas and sectors of health. Once it is a complex subject, the quality evaluation has different di-
mensions and the ones which are the most used by the health services are those that involve structure, processes and results. We aimed at refl ecting on the quality and use of evaluation criteria in the scope of newborn assistance. It is a matter of
bibliographic review on the subject, on a theorical-refl exive perspective. The quality of attention to the new-born is based
on the history of the new-born assistance institutionalization, on the health services structure, human resources, materials
and technological procedures. There are different perspectives of evaluating the new-born assistance quality, including the
use of the mortality index as well as and other specifi c assistance indexes. The offered discussion shows the use of evaluative
practices importance and necessity of implementation, in the search for quality health services in neonatology. KEYWORDS: Health assistance quality; Health evaluation; Health assistance; Infant, Newborn. La calidad abarca diversos sectores y áreas en la salud. Por ser un tema complejo, la evaluación de la calidad tiene diferentes
dimensiones, siendo las más utilizadas por los servicios de salud, las que abarcan estructura, proceso y resultado. Se planteó
refl exionar sobre la calidad y el uso de sus criterios de evaluación en el ámbito de atención al recién nacido. Se trata de una
revisión bibliográfi ca, en una perspectiva teórico/ refl exiva. Artigo de Refl exão Artigo de Refl exão 2 Enfermeira. Doutora em Enfermagem em Saúde Pública pela Universidade de São Paulo. Professora adjunta da Universidade Federal de Mato Grosso, coor-
denadora do Curso de Mestrado em Enfermagem-UFMT, coordenadora do Comitê de Ética em Pesquisa do Hospital Universitário Júlio Muller. Membro do
Grupo de Pesquisa ARGOS da Faculdade de Enfermagem da UFMT. Endereço: Avenida Fernando Correa s/n. Bairro Coxipó, Cuiabá – Mato Grosso-Brasil. CEP:
78100-000. Email: mamgaiva@yahoo.com.br. 1 Enfermeira. Mestranda do curso de Enfermagem da Universidade Federal de Mato Grosso. Membro do Grupo de Pesquisa ARGOS da Faculdade de Enferma-
gem da UFMT. Endereço: Rua Alves de Oliveira, 1870, Cristo Rei. Várzea Grande – Mato Grosso-Brasil. CEP: 78118-080. Fone/fax: (65) 3615-8805. E-mail:
kim_ufmt@yahoo.com.br INTRODUÇÃO visto que atualmente é o modelo mais utilizado em avalia-
ção dos serviços de saúde. Esse modelo propõe estudos nas dimensões da es-
trutura, processo e resultado. A estrutura implica caracte-
rísticas relativamente estáveis das instituições como: área
física, recursos humanos, materiais, fi nanceiros e a estru-
tura organizacional; o processo refere-se ao conjunto de
atividades desenvolvidas na produção de bens e serviços e,
no setor saúde, nas relações estabelecidas entre profi ssio-
nais e clientes, desde a busca pelo diagnóstico, terapêutica
e assistência adequados. O resultado é a obtenção das ca-
racterísticas desejáveis dos produtos ou serviços, retrata
os efeitos da assistência à saúde do cliente, ou seja, em que
condições o paciente deixa o hospital e se suas expectati-
vas foram atendidas, incluindo aqui pesquisas sobre per-
cepção dos usuários acerca do atendimento dos serviços
de saúde(3). A busca pela qualidade nos serviços de saúde cons-
titui uma preocupação mundial, inicialmente observada
nos países desenvolvidos, expandindo-se nas últimas dé-
cadas para os demais países, tendo sido tema freqüente no
Brasil. A qualidade é um conjunto de atributos que deter-
minam a natureza de algo, diz respeito à superioridade e
excelência(1). Os serviços hospitalares brasileiros são respon-
sáveis por 40% dos custos em saúde no país(2), por este
motivo, espera-se que os custos hospitalares sejam bem
empregados e contribuam para uma assistência de qua-
lidade. Nessas instituições, em geral, concentram-se alta
densidade tecnológica, composta de materiais, equipa-
mentos e diversos profi ssionais especializados, possibili-
tando o acesso da população a diagnósticos e assistência
terapêutica. Quando se pretende estudar qualidade dos serviços
através do modelo de Donabedian, é possível utilizar cri-
térios quantitativos, geralmente aplicados na dimensão de
estrutura e resultados, ou até mesmo critérios qualitativos
aplicados à dimensão dos processos, amparando-se nas
relações entre usuários e prestadores de serviços de saú-
de. A partir dessa perspectiva são possíveis diversas formas
de avaliação. A assistência hospitalar ao recém-nascido é com-
plexa, devido às vulnerabilidades próprias da faixa etária,
necessitando muitas vezes de tecnologias sofi sticadas com
custo elevado. No entanto, pouco se conhece a respeito
do acesso de qualidade dos pacientes a esses serviços,
da disponibilidade dos recursos humanos e materiais
adequados e da efi cácia apresentada em virtude do uso
dos processos da qualidade. INTRODUÇÃO Dessa forma, a avaliação da
qualidade da atenção ao neonato no âmbito hospitalar se
faz necessária, quando se pretende diminuir o índice de
mortalidade neonatal, alcançando também níveis satisfa-
tórios de assistência. Para desenvolver a refl exão do Modelo ora descrito,
foi necessário o uso pertinente de leituras sobre as formas
de avaliar a qualidade nos serviços de saúde, relacionan-
do-os com a assistência neonatal. QUALIDADE NA ATENÇÃO HOSPITALAR AO RECÉM-NASCIDO
QUALITY IN THE HOSPITAL ATTENTION TO THE NEWBORN
CALIDAD EN LA ATENCIÓN DEL HOSPITAL AL RECIÉN NACIDO La calidad de la atención neonatal es resultante de la historia
de la asistencia al recién nacido, de la estructura de los servicios, de los recursos humanos, de los materiales y procedimien-
tos tecnológicos. Hay diferentes maneras de evaluar la calidad de la atención neonatal, incluyendo el uso del indicador de
mortalidad e indicadores específi cos de la asistencia. Las discusiones ofrecidas muestran la importancia y la necesidad de
implementar prácticas evaluadoras, en la búsqueda de servicios/asistencia de calidad en neonatología. p
p
q
g
PALABRAS CLAVE: Calidad de la asistencia a la salud; Evaluación en salud; Asistencia a la salud; Recién nacido. 1 Enfermeira. Mestranda do curso de Enfermagem da Universidade Federal de Mato Grosso. Membro do Grupo de Pesquisa ARGOS da Faculdade de Enferma-
gem da UFMT. Endereço: Rua Alves de Oliveira, 1870, Cristo Rei. Várzea Grande – Mato Grosso-Brasil. CEP: 78118-080. Fone/fax: (65) 3615-8805. E-mail:
kim_ufmt@yahoo.com.br
2 Enfermeira. Doutora em Enfermagem em Saúde Pública pela Universidade de São Paulo. Professora adjunta da Universidade Federal de Mato Grosso, coor-
denadora do Curso de Mestrado em Enfermagem-UFMT, coordenadora do Comitê de Ética em Pesquisa do Hospital Universitário Júlio Muller. Membro do
Grupo de Pesquisa ARGOS da Faculdade de Enfermagem da UFMT. Endereço: Avenida Fernando Correa s/n. Bairro Coxipó, Cuiabá – Mato Grosso-Brasil. CEP:
78100-000. Email: mamgaiva@yahoo.com.br. 2 Enfermeira. Doutora em Enfermagem em Saúde Pública pela Universidade de São Paulo. Professora adjunta da Universidade Federal de Mato Grosso, coor-
denadora do Curso de Mestrado em Enfermagem-UFMT, coordenadora do Comitê de Ética em Pesquisa do Hospital Universitário Júlio Muller. Membro do
Grupo de Pesquisa ARGOS da Faculdade de Enfermagem da UFMT. Endereço: Avenida Fernando Correa s/n. Bairro Coxipó, Cuiabá – Mato Grosso-Brasil. CEP:
78100-000. Email: mamgaiva@yahoo.com.br. 159 Rev. Rene. Fortaleza, v. 10, n. 1, p. 159-165, jan./mar.2009 Rev. Rene. Fortaleza, v. 10, n. 1, p. 159-165, jan./mar.2009 Artigo de Refl exão QUALIDADE E AVALIAÇÃO DA ATENÇÃO
HOSPITALAR O presente artigo tem por objetivo refl etir sobre a
qualidade e o uso de seus critérios avaliativos no âmbito da
assistência neonatal. Os estudos relativos aos conceitos e processos da
qualidade, possuíram sua construção histórica nos Esta-
dos Unidos (EUA), quando no início do século XX, o Co-
légio de Cirurgiões Americanos passou a assumir a res-
ponsabilidade de avaliar a qualidade dos hospitais e das
práticas cirúrgicas da época. As iniciativas ocorridas nessa
área contribuíram para padronização dos serviços hospi-
talares, através de critérios avaliativos que repercutiram
na melhoria, a nível estrutural, de diversos hospitais nos PERCURSO METODOLÓGICO Trata-se de uma revisão de literatura, teórico-re-
fl exiva, acerca da qualidade e de seus critérios avaliativos
no âmbito da assistência neonatal. O referencial teórico
utilizado para discorrer acerca da qualidade e dos crité-
rios utilizados no processo de avaliação das instituições de
saúde, foi desenvolvido pelo pediatra Avedis Donabedian, Rev. Rene. Fortaleza, v. 10, n. 1, p. 159-165, jan./mar.2009 160 Artigo de Refl exão EUA e que hoje infl uenciam, de certa maneira, a avaliação
dos serviços em saúde, na busca por uma assistência de
qualidade(4). gia e radiologia. Mas, é no âmbito hospitalar onde é atu-
almente mais utilizado, por se tratar de um ambiente que
exige estruturação complexa, concentração de recursos de
diagnóstico e tratamento, além de ofertar assistência de
saúde continuada(9). Os princípios defendidos por Donabedian encon-
tram-se incorporados aos processos de avaliação da qua-
lidade no Brasil, propostos pelo Ministério da Saúde (MS)
(5). De acordo com o Manual de Acreditação do MS, os
padrões e indicadores aplicados na avaliação de hospitais
“procuram avaliar dentro de um único serviço aspectos de
estrutura, processo e resultado, procurando indicadores
dinâmicos destes padrões e que refl itam a qualidade das
prestações de serviço” (5:13). A acreditação é defi nida como um procedimento de
avaliação dos recursos institucionais que se dá de forma
voluntária, periódica e reservada(5). É uma ação coorde-
nada por uma organização ou agência não governamental,
externa à instituição e encarregada do desenvolvimento e
atualização da metodologia de acreditação. Seus princípios
têm um caráter eminentemente educativo, voltado para a
melhoria contínua do serviço(9). O setor saúde no Brasil vem trabalhando com ava-
liação hospitalar desde a década de 70, sem que houves-
se impacto sobre a qualidade dos serviços prestados(6). Observam-se investimentos ainda em estudos de avaliação
da estrutura, utilizando dados quantitativos. Em outros pa-
íses, como os EUA, a maioria dos hospitais já ultrapassou o
estudo da estrutura e dos procedimentos clínicos, concen-
trando atualmente seus esforços na análise de resultados
da assistência prestada(7). As principais vantagens da acreditação são: seguran-
ça para os pacientes e profi ssionais, melhoria da qualida-
de da assistência e construção de equipe capacitada. Além
disso, é um útil instrumento de gerenciamento, incluindo
critérios e objetivos concretos adaptados à realidade bra-
sileira. Logo, contribui para a qualifi cação contínua dos
serviços de saúde. PERCURSO METODOLÓGICO Os principais interessados por esse pro-
cesso são: organizações de saúde; administradores; profi s-
sionais de saúde; o próprio governo e os usuários(9). Atualmente, a maioria dos estudos brasileiros sobre
qualidade em saúde pauta-se nos modelos avaliativos tra-
dicionais, com foco nas dimensões objetiváveis, excluindo
a dimensão intersubjetiva propriamente humana. Cabe as-
sinalar que ambas correspondem a dimensões inerentes a
fenômenos complexos como a saúde. Não se pode defen-
der o predomínio de uma sobre a outra e sim utilizá-las
de forma associada. Recomenda-se, quando se pensa em
avaliação dos serviços de saúde de forma mais ampliada, a
associação de métodos quantitativos e qualitativos, permi-
tindo maior rigor avaliativo(8). A experiência de países pioneiros nesse assunto,
como os EUA, mostrou que o processo de acreditação
contribui para mudança e qualidade dos serviços. Durante
o processo de preparação, antes de ser submetido à avalia-
ção, percebe-se que os diversos setores do hospital desen-
volvem sua capacidade de comunicação, permitindo uma
auto-avaliação, discussão e correção de falhas, visando
atingir os padrões considerados de qualidade(2). A partir de experiências na avaliação de hospitais
no Brasil, foi elaborado em 1998, pelo Ministério da
Saúde, o Manual de Acreditação que traz os padrões a
serem avaliados, servindo como um guia para as institui-
ções que desejam ser acreditadas. O processo de avalia-
ção fundamenta-se nos seguintes padrões: organização
da assistência médica, diagnóstico e terapêutica, apoio
técnico, incluindo neste o serviço de enfermagem e ou-
tros profi ssionais não médicos, processamento e abaste-
cimento da instituição, relacionada à lavanderia e Centro
de Materiais Estéreis (CME) e, apoio administrativo, in-
cluindo aspectos administrativos, estruturais e de insta-
lações. Dessa maneira, cada um dos padrões descreve A qualidade da atenção nos serviços de saúde é uma
questão complexa, com implicações específi cas, que tem
por fi nalidade avaliar programas criados e aperfeiçoados
por instituições de saúde, visando “acreditar” essas orga-
nizações. A acreditação é um dos métodos propostos para
avaliar essa qualidade. O processo de acreditação passou a ser discutido
e difundido no Brasil no início da década de 90, podendo
ser realizado em diversos níveis dos serviços de saúde: am-
bulatorial, domiciliar, hemoterapia, laboratórios, nefrolo- Rev. Rene. Fortaleza, v. 10, n. 1, p. 159-165, jan./mar.2009 Rev. Rene. Fortaleza, v. 10, n. 1, p. 159-165, jan./mar.2009 161 Artigo de Refl exão quesitos importantes e relacionados aos diversos setores
e serviços do hospital(5). PERCURSO METODOLÓGICO modernas UTIN possibilitaram a sobrevida de crianças até
então consideradas inviáveis devido à prematuridade e ao
baixo peso de nascimento(11). A Organização Nacional de Acreditação (ONA) é o
órgão responsável em avaliar a qualidade dos serviços de
saúde no Brasil. A ONA foi reconhecida pelo Ministério da
Saúde, através da portaria GM/MS nº 538 de 2001, como
instituição competente e autorizada a operacionalizar o de-
senvolvimento do processo de Acreditação Hospitalar(1). À medida que a medicina continua a desenvolver
tecnologias para a manutenção da vida, muitos recém-
nascidos de alto risco, que no passado teriam morrido al-
gumas horas ou dias após o nascimento, hoje sobrevivem. Pode-se dizer que o aumento na sobrevida neonatal estaria
relacionado também a uma assistência estruturada e de
qualidade, sendo um estímulo para continuar investindo
nessa direção. De acordo com dados atuais da ONA, nota-se que
poucos hospitais vêm submetendo-se ao processo de acre-
ditação. Em consulta relativa a 2008, apenas 86 serviços
hospitalares constam no sistema com o título de acredi-
tação – destes 21 foram considerados acreditados, 38
acreditados pleno, e apenas 27 acreditados por excelên-
cia. Observa-se que a maioria das instituições hospitalares
acreditadas concentra-se na região sudeste(9). Nesse sen-
tido, é necessária uma divulgação ampla em todo país de
como se dá o processo de avaliação e da sua importância
para o hospital como uma forma de melhoria da atenção
à saúde. A assistência ao recém-nascido apresenta-se de for-
ma complexa, com cuidados indispensáveis à sua saúde,
de modo que as relações e os processos assistenciais sejam
benéfi cos tanto para o neonato quanto para sua família; e
que os resultados sejam alcançados. Quando se considera
a qualidade assistencial, pautada no modelo de Donabe-
dian, faz-se necessário que os serviços possuam uma es-
trutura com espaço físico, recursos humanos e materiais,
tecnologias de diagnóstico e terapêutica em condições e
quantidades adequadas para assistir ao recém-nascido. Existem diversas formas de avaliar qualidade em saúde e
essa diversidade também se aplica às avaliações no âmbito
da assistência neonatal. Qualidade e avaliação da atenção hospitalar ao
recém-nascido Atualmente, a assistência hospitalar prestada ao
recém-nascido envolve uma estrutura física diferenciada,
pautada em vários espaços e níveis de cuidados, que se
iniciam no Centro Obstétrico e são complementados pelo
Alojamento Conjunto e pela Unidade de Terapia Intensiva
Neonatal. Compreende-se que o processo de organização
desses serviços no âmbito hospitalar contribuiu para defi -
nição de ambientes assistenciais adequados à condição do
neonato, seja ele de baixo risco, atendido no alojamento
conjunto, ou de alto risco, atendido nas UTIN. Esses am-
bientes foram estruturados de forma a prestar uma melhor
assistência a essas crianças em um momento em que estão
mais vulneráveis. A institucionalização da assistência ao neonato ini-
ciou-se, no fi m do século XIX, com a criação dos primeiros
berçários para prematuros, que tinham um atendimento
centrado na prevenção de infecções, controle da tempe-
ratura e alimentação. Esses berçários foram criados com
a fi nalidade de reduzir a mortalidade neonatal nos países
industrializados. No início do século XX, a assistência nes-
ses berçários estendeu-se para os demais recém-nascidos,
face à alta taxa de mortalidade infantil por diarréia e infec-
ções respiratórias(10). Aos poucos os clássicos berçários
foram sendo substituídos pelas unidades de Alojamento
Conjunto, por Unidades de Cuidados Intermediários Neo-
natais (UCIN) e por Unidades de Terapia Intensiva Neona-
tal (UTIN). Além da estrutura física como um dos determinantes
para uma assistência neonatal de qualidade, é fundamental
o investimento em profi ssionais especializados, buscando
atender as reais necessidades do neonato e de sua família,
pois independente da área de atuação, o fator humano,
responsável pela assistência ao paciente, é imprescindível Nas últimas décadas do século XX, houve um incre-
mento na tecnologia de assistência médica hospitalar e,
em especial, na atenção ao neonato prematuro e de mui-
to baixo peso ao nascer. A organização e implantação das Rev. Rene. Fortaleza, v. 10, n. 1, p. 159-165, jan./mar.2009 162 Artigo de Refl exão primeiro e quinto minuto, os quais são dois dos diversos
indicadores possíveis e relacionados à qualidade da assis-
tência neonatal no âmbito hospitalar e que podem ser uti-
lizados pelos serviços neonatais como avaliação “pontual”
da assistência(13). quando se pensa em alcançar bons resultados em saúde. Qualidade e avaliação da atenção hospitalar ao
recém-nascido Para tanto, o Ministério da Saúde elaborou algumas nor-
matizações e recomendações quanto ao número de profi s-
sionais necessários, principalmente em relação às unida-
des de terapia intensiva, as quais exigem maior diversidade
de profi ssionais e especialidade dos mesmos(12). Também encontramos estudos que associam óbitos
perinatais com o processo assistencial ao parto e nasci-
mento, com um grande potencial de produzir modifi ca-
ções na mortalidade fetal, neonatal e infantil. Podemos
citar o estudo(16) quantitativo realizado em 15 materni-
dades brasileiras, que comprovou determinadas variáveis
como indicadores de qualidade de processo relacionados
ao recém-nascido como: resultado do apagar, assistência
pediátrica na sala de parto e característica do serviço neo-
natal que o hospital tem em sua estrutura. A qualidade da atenção neonatal também envolve
a aplicação de tecnologias assistenciais, visando alcançar
menores índices de mortalidade neonatal. Nesse sentido,
a utilização de tecnologias diagnósticas e terapêuticas
específi cas na área neonatal faz parte das determinações
recomendadas pelos órgãos regulamentadores, como o
Ministério da Saúde e algumas organizações profi ssionais
em saúde(13). Logo, o uso de critérios avaliativos como
estrutura física, recursos humanos e tecnologias de diag-
nóstico/terapêutica constituem avaliação da estrutura dos
serviços. De acordo com o estudo acima, se encontrou uma
relação importante entre o tipo de hospital e óbitos perina-
tais. O hospital considerado do tipo I (sem UCIN ou UTIN)
apresentou-se como risco para o óbito perinatal, além dis-
so, nesses hospitais foi encontrado alto índice de nascidos
vivos (38,1%) que não receberam assistência de pediatra
na sala de parto, em contraste com os outros hospitais que
tinham UCIN ou UTIN que apresentaram apenas 2,5% e;
2,2% de nascidos vivos, que não receberam assistência de
pediatra na sala de parto, respectivamente(16). A avaliação da assistência neonatal pode ser feita
através da aplicação de indicadores de mortalidade, o qual
é um importante instrumento de avaliação em saúde, tanto
de cobertura, de qualidade e de resolubilidade dos servi-
ços quanto de programas, de ações específi cas ou ainda
tecnológicas(14). Os estudos sobre mortalidade neonatal na literatura
brasileira são comuns, seja através do cálculo de coefi -
ciente ou estudos que abordem as causas de morte, ou
até mesmos de fatores de risco que possam levar ao óbito
neonatal. A maioria dos estudos deixa claro que a mortali-
dade neonatal relaciona-se intimamente com a qualidade
da assistência materno-infantil. Qualidade e avaliação da atenção hospitalar ao
recém-nascido É importante destacar que a avaliação da atenção
neonatal também pode ser realizada através do processo
de acreditação. A acreditação de um hospital envolve toda
a instituição, da lavanderia à Unidade de Terapia Intensiva,
logo inclui a assistência hospitalar ao neonato. De acordo
com o manual de Acreditação do MS, o item que se refere à
avaliação da unidade de neonatologia dentro dos hospitais
inclui desde aspectos estruturais da unidade à aplicação
de sistema de aferição da qualidade, através de processos
avaliativos internos ou integrados ao hospital. Pode-se res-
saltar que a mobilização do próprio serviço ao elaborar
internamente formas de acompanhar as condições de sua
assistência, permite assim, avaliação e superação dos pro-
blemas encontrados, iniciando mudanças já no momento
pré-acreditação(5). Estudo realizado em uma maternidade pública de
nível terciário de Campinas, São Paulo, demonstrou que
o coefi ciente de mortalidade neonatal no serviço apresen-
tou modifi cações ao longo de uma década e as causas de
morte neonatais evitáveis diminuíram. Os autores dessa
pesquisa associam os resultados atuais às mudanças ocor-
ridas na qualidade da assistência neonatal prestada nesta
maternidade(15). Existem também outros indicadores específi cos de
qualidade da assistência neonatal, os quais podem ser uti-
lizados em pesquisas de avaliação da qualidade. Têm-se
como exemplos, a existência do alojamento conjunto, com
estímulo ao aleitamento materno e o registro de APGAR no A assistência hospitalar ao neonato de risco utiliza
grande aparato tecnológico e profi ssionais especializados
concentrados numa UTIN, considerado um local ideal Rev. Rene. Fortaleza, v. 10, n. 1, p. 159-165, jan./mar.2009 Rev. Rene. Fortaleza, v. 10, n. 1, p. 159-165, jan./mar.2009 163 Artigo de Refl exão para manter a sobrevida dessas crianças. Entretanto, nas
maternidades dos países subdesenvolvidos e em desen-
volvimento, há evidente escassez de recursos tecnológicos
e humanos, infra-estrutura inadequada e alta incidência
de infecção hospitalar em decorrência da superlotação
hospitalar(17). tam assistência intensiva neonatal/pediátrica e ao parto,
também levantam problemas que permeiam a atenção ao
neonato, contribuindo para a avaliação da qualidade nas
instituições pesquisadas, além de servirem de suporte para
outros estudos avaliativos nessa área(16,18). Outra possibilidade de avaliação da assistência neo-
natal pode ser realizada com foco no resultado, incluindo
então a satisfação ou até mesmo a resposta do usuário a
assistência recebida. Qualidade e avaliação da atenção hospitalar ao
recém-nascido Embora algumas pesquisas desse tipo
sejam de natureza quantitativa, não se pode excluir o cará-
ter subjetivo encontrado em pesquisas que avaliam a acei-
tação de programas ou de serviços específi cos. A exemplo
disso tem-se uma pesquisa realizada em uma maternidade
de Fortaleza-Ceará, onde um dos objetivos era avaliar o
conhecimento das mães acerca dos benefícios do Método
Mãe Canguru para o binômio mãe-fi lho. Além de desvendar
resultados interessantes sobre os sentimentos maternos vi-
venciados nesse ambiente de cuidado neonatal, o referido
estudo possibilitou apreender o nível de satisfação dessas
mães pela assistência recebida, através de relatos positivos
apresentados pela maioria delas(17). Na realidade brasileira, apesar de toda regulamen-
tação existente acerca da assistência neonatal, as UTIN,
as unidades de cuidados intermediários e os alojamentos
conjuntos têm difi culdades em atender as exigências míni-
mas preconizadas pelo MS e muitas funcionam de manei-
ra precária. Além disso, muito pouco se conhece sobre a
distribuição, estrutura e organização dessas unidades, nas
diferentes regiões do Brasil. No que refere à avaliação da estrutura das UTIN,
pode-se citar um estudo realizado em 2003 que analisa o
acesso e estrutura das UTI pediátricas e neonatais de São
Paulo, partindo dos princípios de avaliação propostos por
Donabedian. Os dados relativos à área física e instalações
mostram que se convive com unidades que fi cam aquém
das exigências feitas pelas normatizações brasileiras. Ape-
sar de a maioria das UTI informarem ter acesso a todas as
tecnologias necessárias de apoio diagnóstico e terapêuti-
co, alguns materiais e equipamentos essenciais não foram
encontrados, por exemplo, o manguito do esfi gnomanô-
metro utilizado para aferir a pressão arterial(2). Independentemente da diversidade dos processos
avaliativos, o que se espera é que os serviços hospitala-
res prestadores de assistência ao neonato compreendam
a importância das ferramentas de avaliação e da sua con-
tribuição na busca dos resultados, ou seja, a qualidade da
assistência. Outro estudo que segue neste caminho avaliativo foi
realizado em 12 maternidades do Estado de São Paulo inte-
gradas ao Sistema Único de Saúde, buscando analisar não
apenas indicadores de estrutura como também de proces-
so, envolvendo a assistência ao parto e ao recém-nascido. Foram encontrados resultados insatisfatórios, como pre-
sença de médico pediatra e enfermeiro para atendimento
neonatal na sala de parto (24 horas/dia) apenas em uma
das maternidades. Qualidade e avaliação da atenção hospitalar ao
recém-nascido Em relação a indicadores de proces-
so, podemos ressaltar que o contato pele-a-pele mãe/bebê
ocorreu em apenas em seis (4,5%) dos 128 partos ob-
servados. Esses resultados demonstram que a assistência
neonatal não atende critérios fundamentais recomendadas
pelo Ministério da Saúde(18). REFERÊNCIAS 1. Helito RAB. Processos de qualifi cação e avaliação de
serviços de saúde. In: Innocenzo MD. Indicadores, Au-
ditorias e Certifi cações. São Paulo: Martinari, 2006, p. 59-70. 1. Helito RAB. Processos de qualifi cação e avaliação de
serviços de saúde. In: Innocenzo MD. Indicadores, Au-
ditorias e Certifi cações. São Paulo: Martinari, 2006, p. 59-70. 11. Klaus MH, Faranoff AA. Alto Risco em Neonatologia. 4a
ed. Rio de Janeiro: Guanabara Koogan, 1995. 12. Brasil. Ministério da Saúde. Portaria MS/GM 3432, de
12 de agosto de 1998. Estabelece critérios de classifi -
cação para as unidades de tratamento intensivo. Brasí-
lia (DF); 1998. 2. Souza DC de. Avaliação da estrutura das unidades de
terapia intensiva pediátrica e neonatal no município
de São Paulo. [dissertação de Mestrado em Medicina]. São Paulo (SP): Faculdade de Medicina. Universidade
de São Paulo, 2003. 206f. 13. Magalhães M da C. Atenção hospitalar perinatal e a
mortalidade neonatal no município de Juiz de Fora –
MG [dissertação de Mestrado em Saúde Pública]. Rio
de Janeiro (RJ): Escola Nacional de Saúde Pública:
Fundação Oswaldo Cruz; 2000. 203f. 3. Donabedian A. The quality of care. How can it be asses-
sed? EUA 1988; 260(12): 1743-8. 3. Donabedian A. The quality of care. How can it be asses-
sed? EUA 1988; 260(12): 1743-8. 4. Feldman BL. Como alcançar a qualidade nas institui-
ções de saúde: critérios de avaliações, procedimentos
de controle, gerenciamento de riscos hospitalares até
a certifi cação. São Paulo: Martinari, 2004, p. 47-63. 14. Brasil. Ministério da Saúde. Secretaria de Políticas de
Saúde. Análise de mortalidade – Delineamentos bási-
cos. Brasília (DF): 1991. 15. Brenelli – Vitali MA, Castro de R, Pavarin LB. Causas
básicas de morte neonatal em uma maternidade de
nível terciário: mudanças de uma década. Rev. Ciênc. Méd. Campinas (SP) 2003; 12(4): 331-9. 5. Brasil. Ministério da Saúde. Secretaria de políticas de
Saúde, Departamento de Avaliação de Políticas de saú-
de. Vade-mécum da Acreditação Hospitalar Brasileira. Brasília (DF): 1999. 16. Lansky S, França E, César CC, Neto LCM, Leal M do C. Mortes perinatais e avaliação da assistência ao parto
em maternidades do Sistema Único de Saúde em Belo
Horizonte, Minas Gerais, Brasil, 1999. CAD Saúde Pú-
blica, Rio de Janeiro (RJ) 2006; 22 (1): 117-30. 6. Schiesari LMC, Kisil M. A avaliação da qualidade nos
hospitais brasileiros. RAS, São Paulo (SP). 2003;
5(18): 7-17. 7. Adami MP, Maranhão AMSA. CONSIDERAÇÕES FINAIS Discutir a temática qualidade na atenção à saúde
do neonato não implica somente em apresentar concei-
tos, mas em mostrar um conjunto de métodos e estratégias
possíveis de serem aplicados a essa realidade. Nesse senti-
do, esta refl exão apresenta-se como uma possibilidade ini-
cial na discussão da questão, reforçando a necessidade de
se avançar na refl exão em torno dos métodos de avaliação
da qualidade do atendimento ao recém-nascido. Entende-se que a qualidade da atenção ao neonato
passa pela qualifi cação dos recursos humanos, aquisição
e manutenção de materiais e equipamentos, mudanças na
cultura organizacional das instituições, envolvimento e par- De uma forma geral, a maior parte dos estudos de
avaliação da qualidade da assistência ao recém-nascido
os quais descrevem estruturalmente os serviços que pres- Rev. Rene. Fortaleza, v. 10, n. 1, p. 159-165, jan./mar.2009 164 Artigo de Refl exão Acessado em 20 de Julho de 2008. Disponível em:
http://www.onaeducare.org.br/ona/site/internal_insti-
tucional.jsp?pagesite=conheca. ticipação dos usuários no processo assistencial e também
de avaliação, de forma a construir serviços que atendam as
necessidades da população neonatal e de sua família. 10. Kamada I, Scochi CGS, Rocha SMM. Assistência ao
recém-nascido e sua organização a nível hospitalar:
Revisão de Literatura. Rev. Paulista de Hospitais. São
Paulo (SP) 1990; 28(5): 65-73. REFERÊNCIAS Qualidade dos serviços de
saúde: conceitos e métodos avaliativos. Acta Paul. Enf.,
São Paulo (SP) 1995; 8(4): 47-55. 17. Campos ACS, Carvalho MPL, Rolim KMC, Alencar AJC. Vivência no Método Mãe Canguru: percepção da mãe. Rev. Rene, Fortaleza (CE) 2008; 9(3): 28-36. 8. Bosi MLM, Uchimura KY. Avaliação da qualidade ou
avaliação qualitativa em saúde. Rev Saúde Pública.,
São Paulo (SP) 2007; 41(1): 150-3. 18. Parada CMG de LM, Carvalhães MA de BL. Childbir-
th care: contributing to the debate on human deve-
lopment. Rev Latino-Am Enfermagem, Ribeirão Preto
(SP) 2007; 15(spe):792-8. 9. Organização Nacional de Acreditação [homepage na
internet]. Serviços acreditados. [Atualizado em 2008]. RECEBIDO: 15/09/2008
ACEITO: 18/02/2009 RECEBIDO: 15/09/2008
ACEITO: 18/02/2009 RECEBIDO: 15/09/2008
ACEITO: 18/02/2009 Rev. Rene. Fortaleza, v. 10, n. 1, p. 159-165, jan./mar.2009 Rev. Rene. Fortaleza, v. 10, n. 1, p. 159-165, jan./mar.2009 165
|
https://openalex.org/W4251690842
|
https://peerj.com/articles/4007v0.3/submission
|
English
| null |
Peer Review #2 of "The response of soil microbial communities to variation in annual precipitation depends on soil nutritional status in an oligotrophic desert (v0.3)"
| null | 2,017
|
cc-by
| 22,970
|
The response of soil microbial communities to variation in
annual precipitation depends on soil nutritional status in an
oligotrophic desert tiel-González 1 , Yunuen Tapia-Torres 2 , Valeria Souza 3 , Felipe García-Oliva Corresp. 1 1 Instituto de Investigaciones en Ecosistemas y Sustentabilidad, Universidad Nacional Autónoma de México, Morelia, Michoacán, Mexico
2 Escuela Nacional de Estudios Superiores Unidad Morelia, Universidad Nacional Autónoma de México, Morelia, Michoacán, Mexico
3 Instituto de Ecología, Universidad Nacional Autónoma de México, Mexico, Mexico
Corresponding Author: Felipe García-Oliva
Email address: fgarcia@cieco.unam.mx 1 Instituto de Investigaciones en Ecosistemas y Sustentabilidad, Universidad Nacional Autónoma de México, Morelia, Michoacán, Mexico
2 Escuela Nacional de Estudios Superiores Unidad Morelia, Universidad Nacional Autónoma de México, Morelia, Michoacán, Mexico
3 Instituto de Ecología, Universidad Nacional Autónoma de México, Mexico, Mexico
Corresponding Author: Felipe García-Oliva
Email address: fgarcia@cieco.unam.mx Background. Soil microbial communities (SMC) play a central role in the structure and
function of desert ecosystems. However, the high variability of annual precipitation could
results in the alteration of SMC and related biological processes depending on soil water
potential. The nature of the physiological adjustments made by SMC in order to obtain
energy and nutrients remains unclear under different soil resource availabilities in desert
ecosystems. In order to examine this dynamic, the present study examined the effects of
variation in annual precipitation on physiological adjustments by the SMC across two
vegetation-soil systems of different soil organic matter input in an oligotrophic desert
ecosystem. Methods. We collected soil samples in the Cuatro Ciénegas Basin (Mexico)
under two vegetation covers: rosetophylous scrub (RS) and grassland (G), that differ in
terms of quantity and quality of organic matter. Collections were conducted during the
years 2011, 2012, 2013 and 2014, over which a noticeable variation in the annual
precipitation occurred. The ecoenzymatic activity involved in the decomposition of organic
matter, and the concentration of dissolved, available and microbial biomass nutrients,
were determined and compared between sites and years. Results. In 2011, we observed
differences in bacterial taxonomic composition between the two vegetation covers. The
lowest values of dissolved, available and microbial nutrients in both cover types were
found in 2012. The G soil showed higher values of dissolved and available nutrients in the
wet years. Significant positive correlations were detected between precipitation and the
ratios Cmic:Nmic and Cmic:Pmic in the RS soil and Cmic:Pmic and Nmic:Pmic in the G soil. The slopes of the regression with Cmic and Nmic were higher in the G soil and lower in the
RS soil. Manuscript to be reviewed The response of soil microbial communities to variation in
annual precipitation depends on soil nutritional status in an
oligotrophic desert Moreover, the SMC under each vegetation cover were co-limited by different
nutrients and responded to the sum of water stress and nutrient limitation. Discussion. Soil community within both sites (RS and G) may be vulnerable to drought. However, the PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed community of the site with lower resources (RS) is well adapted to acquire P resources by
ecoenzyme upregulation during years with adequate precipitation, suggesting that this
community is resilient after drought occurs. Under the Global Climate Change scenarios for
desert ecosystems that predict reduced annual precipitation and an increased intensity
and frequency of torrential rains and drought events, the soil microbial communities of
both sites could be vulnerable to drought through C and P co-limitation and reallocation of
resources to physiological acclimatization strategies in order to survive. community of the site with lower resources (RS) is well adapted to acquire P resources by
ecoenzyme upregulation during years with adequate precipitation, suggesting that this
community is resilient after drought occurs. Under the Global Climate Change scenarios for
desert ecosystems that predict reduced annual precipitation and an increased intensity
and frequency of torrential rains and drought events, the soil microbial communities of
both sites could be vulnerable to drought through C and P co-limitation and reallocation of
resources to physiological acclimatization strategies in order to survive. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed Manuscript to be reviewed 1
The response of soil microbial communities to variation in annual precipitation depends on soil
2
nutritional status in an oligotrophic desert
3
4
Cristina Montiel-Gonzáleza, Yunuen Tapia-Torresb, Valeria Souzac and Felipe García-Olivaa*. 5
6
aInstituto de Investigaciones en Ecosistemas y Sustentabilidad, Universidad Nacional Autónoma
7
de México, Morelia, Michoacán, México. 8
cmontiel@cieco.unam.mx; fgarcia@cieco.unam.mx
9
10
bEscuela Nacional de Estudios Superiores Unidad Morelia, Universidad Nacional Autónoma de
11
México, Morelia, Michoacán, México. 12
ytapia@enesmorelia.unam.mx
13
14
cInstituto de Ecología, Departamento de Ecología Evolutiva, Universidad Nacional Autónoma de
15
México, México DF, México. 16
souza@unam.mx
17
18
*Corresponding autor: Felipe García-Oliva. Instituto de Investigaciones en Ecosistemas y
19
Sustentabilidad, Universidad Nacional Autónoma de México. Antigua carretera a Pátzcuaro No. 20
8701, col. Ex Hacienda de San José de la Huerta, CP 58190, Morelia, Michoacán, México. 21
fgarcia@cieco.unam.mx, +52 443 322 2715. 22 23 PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 24
Abstract
25
Background. Soil microbial communities (SMC) play a central role in the structure and function
26
of desert ecosystems. However, the high variability of annual precipitation could results in the
27
alteration of SMC and related biological processes depending on soil water potential. The nature
28
of the physiological adjustments made by SMC in order to obtain energy and nutrients remains
29
unclear under different soil resource availabilities in desert ecosystems. In order to examine this
30
dynamic, the present study examined the effects of variation in annual precipitation on
31
physiological adjustments by the SMC across two vegetation-soil systems of different soil
32
organic matter input in an oligotrophic desert ecosystem. 24
Abstract 33
Methods. We collected soil samples in the Cuatro Ciénegas Basin (Mexico) under two
34
vegetation covers: rosetophylous scrub (RS) and grassland (G), that differ in terms of quantity
35
and quality of organic matter. Collections were conducted during the years 2011, 2012, 2013 and
36
2014, over which a noticeable variation in the annual precipitation occurred. The ecoenzymatic
37
activity involved in the decomposition of organic matter, and the concentration of dissolved,
38
available and microbial biomass nutrients, were determined and compared between sites and
39
years. 33
Methods. We collected soil samples in the Cuatro Ciénegas Basin (Mexico) under two
34
vegetation covers: rosetophylous scrub (RS) and grassland (G), that differ in terms of quantity
35
and quality of organic matter. Collections were conducted during the years 2011, 2012, 2013 and
36
2014, over which a noticeable variation in the annual precipitation occurred. The ecoenzymatic
37
activity involved in the decomposition of organic matter, and the concentration of dissolved,
38
available and microbial biomass nutrients, were determined and compared between sites and
39
years. 40
Results. In 2011, we observed differences in bacterial taxonomic composition between the two
41
vegetation covers. The lowest values of dissolved, available and microbial nutrients in both
42
cover types were found in 2012. The G soil showed higher values of dissolved and available
43
nutrients in the wet years. Significant positive correlations were detected between precipitation
44
and the ratios Cmic:Nmic and Cmic:Pmic in the RS soil and Cmic:Pmic and Nmic:Pmic in the G
45
soil. The slopes of the regression with Cmic and Nmic were higher in the G soil and lower in the 40
Results. In 2011, we observed differences in bacterial taxonomic composition between the two
41
vegetation covers. The lowest values of dissolved, available and microbial nutrients in both
42
cover types were found in 2012. The G soil showed higher values of dissolved and available
43
nutrients in the wet years. Significant positive correlations were detected between precipitation
44
and the ratios Cmic:Nmic and Cmic:Pmic in the RS soil and Cmic:Pmic and Nmic:Pmic in the G
45
soil. The slopes of the regression with Cmic and Nmic were higher in the G soil and lower in the PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) 58
Introduction 59
In desert ecosystems, precipitation is highly variable among years and this variability has
60
increased in recent years due to the effect of Global Climate Change (GCC) (Bell et al. 2014;
61
IPCC 2013). The scenarios derived from GCC models for desert ecosystems predict reduced
62
annual precipitation, as well as increases in the annual precipitation variability by the end of the
63
21st century, including an increase in the frequency and intensity of both torrential rain and
64
drought events (Holmgren et al. 2006; IPCC 2013). The high variability of annual precipitation
65
projected for desert ecosystems could alter biological processes dependent on soil water
66
potential, as is the case with the processes related to soil organic matter (SOM) decomposition
67
(D'Odorico & Bhattachan 2012; Fay et al. 2008; Thomey et al. 2011). For example, enzymatic
68
activity stimulated by rainfall in desert ecosystems may result in most of the total annual
69
mineralization that occurs in desert soils (Manzoni et al. 2012). However when soil water
70
potential decreases, the metabolic activity of most soil microbial species is reduced, and thus a
71
decline in nutrient mineralization can occur. Additionally, soil drying reduces enzymatic activity
72
and microbial mobility, which reduces substrate supply for the decomposers (Henry 2013;
73
Manzoni et al. 2012). Likewise, studies in a semiarid region in New Mexico (Cregger et al. 74
2012) and in the Chihuahuan Desert (Bell et al. 2009; Bell et al. 2014) showed that the high
75
precipitation variability significantly altered the structure of the soil microbial community,
76
mainly due to a change in the fungal/bacterial ratio and consequently altered microbial
77
community functional dynamics. 78
Microbial communities play a central role in the structure and functioning of desert ecosystems 59
In desert ecosystems, precipitation is highly variable among years and this variability has
60
increased in recent years due to the effect of Global Climate Change (GCC) (Bell et al. 2014;
61
IPCC 2013). The scenarios derived from GCC models for desert ecosystems predict reduced
62
annual precipitation, as well as increases in the annual precipitation variability by the end of the
63
21st century, including an increase in the frequency and intensity of both torrential rain and
64
drought events (Holmgren et al. 2006; IPCC 2013). Manuscript to be reviewed 46
RS soil. Moreover, the SMC under each vegetation cover were co-limited by different nutrients
47
and responded to the sum of water stress and nutrient limitation. 48
Discussion. Soil community within both sites (RS and G) may be vulnerable to drought. 49
However, the community of the site with lower resources (RS) is well adapted to acquire P
50
resources by ecoenzyme upregulation during years with adequate precipitation, suggesting that
51
this community is resilient after drought occurs. Under the Global Climate Change scenarios for
52
desert ecosystems that predict reduced annual precipitation and an increased intensity and
53
frequency of torrential rains and drought events, the soil microbial communities of both sites
54
could be vulnerable to drought through C and P co-limitation and reallocation of resources to
55
physiological acclimatization strategies in order to survive. 46
RS soil. Moreover, the SMC under each vegetation cover were co-limited by different nutrients
47
and responded to the sum of water stress and nutrient limitation. 46
RS soil. Moreover, the SMC under each vegetation cover were co-limited by different nutrients
47
and responded to the sum of water stress and nutrient limitation. 46
RS soil. Moreover, the SMC under each vegetation cover were co-limited by different nutrients
47
and responded to the sum of water stress and nutrient limitation. 48
Discussion. Soil community within both sites (RS and G) may be vulnerable to drought. 49
However, the community of the site with lower resources (RS) is well adapted to acquire P
50
resources by ecoenzyme upregulation during years with adequate precipitation, suggesting that
51
this community is resilient after drought occurs. Under the Global Climate Change scenarios for
52
desert ecosystems that predict reduced annual precipitation and an increased intensity and
53
frequency of torrential rains and drought events, the soil microbial communities of both sites
54
could be vulnerable to drought through C and P co-limitation and reallocation of resources to
55
physiological acclimatization strategies in order to survive. 57 PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) 58
Introduction The high variability of annual precipitation
65
projected for desert ecosystems could alter biological processes dependent on soil water 78
Microbial communities play a central role in the structure and functioning of desert ecosystems
79
since they represent an important pool of soil C, N, and P. Indeed, it has been suggested that the
80
amount of N and P contained within the soil microorganism biomass is comparable to the N and PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 81
P content within the plant biomass in desert ecosystems (Coleman & Whitman 2005). Moreover
82
microbial communities can help accelerate the transformation of molecules containing C, N, and
83
P by producing soil extracellular enzymes (ecoenzymes) (Sinsabaugh & Follstad Shah 2012;
84
Sinsabaugh et al. 2009) that lead to the fragmentation, depolymerization and mineralization of
85
organic matter (Singh et al. 2014). Microorganisms can only assimilate soluble organic
86
compounds of a molecular weight lower than 1 kDa and must therefore break down, or
87
depolymerize, most of the organic matter molecules (where between 72 and 87% of the DOC in
88
grassland soils is larger than 1 kDa) in order to access the nutrients and energy contained within
89
the organic molecules (Cregger et al. 2012; Farrell et al. 2014; Jones et al. 2012). The
90
microorganisms produce hydrolytic or oxidative ecoenzymes that degrade organic matter,
91
producing assimilable dissolved organic nutrients that are rapidly immobilized within their
92
biomass (Conant et al. 2011; Sinsabaugh & Follstad Shah 2012). Additionally, in desert
93
ecosystems, the natural distribution of different vegetation types can produce spatial
94
heterogeneity in the quantity and quality of organic matter (Austin et al. 2004; Housman et al. 95
2007). In these ecosystems, the depolymerization process will therefore require the production o
96
different ecoenzymes, since the organic matter under each vegetation type contains a particular
97
combination of structurally simple and complex molecules that promote differences in the soil
98
nutrient dynamics mediated by the microbial community (Conant et al. 2011). However, the
99
complete organic matter decomposition process requires a chain of enzymatic reactions where
100
each ecoenzyme acts on a different substrate and is produced by different microbial groups
101
(Ekschmitt et al. 2005). Additionally, the soil microbial communities can exhibit functional
102
redundancy in the ecoenzyme production (Allison & Martiny 2008). 81
P content within the plant biomass in desert ecosystems (Coleman & Whitman 2005). Moreover,
82
microbial communities can help accelerate the transformation of molecules containing C, N, and
83
P by producing soil extracellular enzymes (ecoenzymes) (Sinsabaugh & Follstad Shah 2012;
84
Sinsabaugh et al. 2009) that lead to the fragmentation, depolymerization and mineralization of
85
organic matter (Singh et al. 2014). Manuscript to be reviewed This high demand for energy (C) and nutrients (N and P) can be offset by
108
reallocation of these resources, generating a trade-off in which the microbial community invests
109
C, N and P in either growth or survival (Evans & Wallenstein 2012; Schimel et al. 2007). Some
110
consequences of such resource redirection are: 1) a limited production of ecoenzymes for
111
nutrient acquisition (i.e. for SOM decomposition) (Burns et al. 2013; Henry 2013; Steinweg et al. 112
2013) and 2) reduced growth of the microbial community (i.e. decreased protein synthesis)
113
(Schimel et al. 2007). Resource reallocation increases the vulnerability of some microbial groups
114
that produce a change in the structure and function of the soil microbial community also
115
affecting the energy flow (C) and nutrient dynamics of N and P at the ecosystem level (Esch et
116
al. 2016; Evans & Wallenstein 2012; Thibault & Brown 2008). This variability strongly affects
117
microbial community development in resource-limited environments, because the adaptation
118
rates of microbial species are constrained by the resource cost of physiological adjustment
119
(Wallenstein & Hall 2012). Wallenstein and Hall (2012) proposed that sites limited by nutrients
120
are more vulnerable to annual rainfall variability, because the microbial community must invest
121
energy in nutrient acquisition, and consequently reducing its capacity for adaptation required by
122
fluctuation in water availability. Sites with low resource availability could be therefore more
123
vulnerable to annual precipitation variability. 124
The Chihuahuan desert has been classified as one of the most biologically outstanding habitats 103
Soil microorganisms have developed mechanisms of physiological acclimatization to cope with
104
precipitation variability (Schimel & Schaeffer 2012). These mechanisms generate physiological
105
costs for the microbial community that derive from the need for high investments of energy (C)
106
and nutrients (N and P) in order to survive (Classen et al. 2015; Schimel et al. 2007; Schimel &
107
Schaeffer 2012). This high demand for energy (C) and nutrients (N and P) can be offset by
108
reallocation of these resources, generating a trade-off in which the microbial community invests
109
C, N and P in either growth or survival (Evans & Wallenstein 2012; Schimel et al. 2007). Some
110
consequences of such resource redirection are: 1) a limited production of ecoenzymes for
111
nutrient acquisition (i.e. for SOM decomposition) (Burns et al. 2013; Henry 2013; Steinweg et al. Manuscript to be reviewed Microorganisms can only assimilate soluble organic
86
compounds of a molecular weight lower than 1 kDa and must therefore break down, or
87
depolymerize, most of the organic matter molecules (where between 72 and 87% of the DOC in
88
grassland soils is larger than 1 kDa) in order to access the nutrients and energy contained within
89
the organic molecules (Cregger et al. 2012; Farrell et al. 2014; Jones et al. 2012). The
90
microorganisms produce hydrolytic or oxidative ecoenzymes that degrade organic matter,
91
producing assimilable dissolved organic nutrients that are rapidly immobilized within their
92
biomass (Conant et al. 2011; Sinsabaugh & Follstad Shah 2012). Additionally, in desert
93
ecosystems, the natural distribution of different vegetation types can produce spatial
94
heterogeneity in the quantity and quality of organic matter (Austin et al. 2004; Housman et al. 95
2007). In these ecosystems, the depolymerization process will therefore require the production of
96
different ecoenzymes, since the organic matter under each vegetation type contains a particular
97
combination of structurally simple and complex molecules that promote differences in the soil
98
nutrient dynamics mediated by the microbial community (Conant et al. 2011). However, the
99
complete organic matter decomposition process requires a chain of enzymatic reactions where
100
each ecoenzyme acts on a different substrate and is produced by different microbial groups
101
(Ekschmitt et al. 2005). Additionally, the soil microbial communities can exhibit functional
102
redundancy in the ecoenzyme production (Allison & Martiny 2008). PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 103
Soil microorganisms have developed mechanisms of physiological acclimatization to cope with
104
precipitation variability (Schimel & Schaeffer 2012). These mechanisms generate physiological
105
costs for the microbial community that derive from the need for high investments of energy (C)
106
and nutrients (N and P) in order to survive (Classen et al. 2015; Schimel et al. 2007; Schimel &
107
Schaeffer 2012). This high demand for energy (C) and nutrients (N and P) can be offset by
108
reallocation of these resources, generating a trade-off in which the microbial community invests
109
C, N and P in either growth or survival (Evans & Wallenstein 2012; Schimel et al. 2007). Some
110
consequences of such resource redirection are: 1) a limited production of ecoenzymes for
111
nutrient acquisition (i.e. for SOM decomposition) (Burns et al. 2013; Henry 2013; Steinweg et al. 112
2013) and 2) reduced growth of the microbial community (i.e. decreased protein synthesis)
113
(Schimel et al. 2007). Resource reallocation increases the vulnerability of some microbial groups
114
that produce a change in the structure and function of the soil microbial community also
115
affecting the energy flow (C) and nutrient dynamics of N and P at the ecosystem level (Esch et
116
al. 2016; Evans & Wallenstein 2012; Thibault & Brown 2008). This variability strongly affects
117
microbial community development in resource-limited environments, because the adaptation
118
rates of microbial species are constrained by the resource cost of physiological adjustment
119
(Wallenstein & Hall 2012). Wallenstein and Hall (2012) proposed that sites limited by nutrients
120
are more vulnerable to annual rainfall variability, because the microbial community must invest
121
energy in nutrient acquisition, and consequently reducing its capacity for adaptation required by
122
fluctuation in water availability. Sites with low resource availability could be therefore more
123
vulnerable to annual precipitation variability. 124
The Chihuahuan desert has been classified as one of the most biologically outstanding habitats
125
globally by the World Wildlife Fund (Archer & Predick 2008) The Cuatro Ciénegas Basin 103
Soil microorganisms have developed mechanisms of physiological acclimatization to cope with
104
precipitation variability (Schimel & Schaeffer 2012). These mechanisms generate physiological
105
costs for the microbial community that derive from the need for high investments of energy (C)
106
and nutrients (N and P) in order to survive (Classen et al. 2015; Schimel et al. 2007; Schimel &
107
Schaeffer 2012). Manuscript to be reviewed 112
2013) and 2) reduced growth of the microbial community (i.e. decreased protein synthesis)
113
(Schimel et al. 2007). Resource reallocation increases the vulnerability of some microbial groups
114
that produce a change in the structure and function of the soil microbial community also
115
affecting the energy flow (C) and nutrient dynamics of N and P at the ecosystem level (Esch et
116
al. 2016; Evans & Wallenstein 2012; Thibault & Brown 2008). This variability strongly affects
117
microbial community development in resource-limited environments, because the adaptation
118
rates of microbial species are constrained by the resource cost of physiological adjustment
119
(Wallenstein & Hall 2012). Wallenstein and Hall (2012) proposed that sites limited by nutrients
120
are more vulnerable to annual rainfall variability, because the microbial community must invest
121
energy in nutrient acquisition, and consequently reducing its capacity for adaptation required by
122
fluctuation in water availability. Sites with low resource availability could be therefore more
123
vulnerable to annual precipitation variability. 124
The Chihuahuan desert has been classified as one of the most biologically outstanding habitats 124
The Chihuahuan desert has been classified as one of the most biologically outstanding habitats
125
globally by the World Wildlife Fund (Archer & Predick 2008). The Cuatro Ciénegas Basin PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 126
(CCB), which is the study site of the present investigation, is part of the Chihuahuan desert and
127
is considered the most important wetland of Mexico for its high levels of endemism and
128
biodiversity (Souza et al. 2011). Moreover, the CCB has been listed as an ultra-oligotrophic site
129
due to low P concentrations in the water and soil, which can constitute a strong potential for P
130
limitation of microbial growth (Elser et al. 2005; Tapia-Torres et al. 2015a). A study in the CCB
131
desert reported that, in the same soil type with different vegetation cover (grassland and desert
132
scrub) differences in OM content promotes variation in DOC concentration, which represents the
133
main energy source for soil microorganisms (Tapia-Torres et al. 2015b). The higher DOC
134
concentration under grassland soil compared to desert scrub soil favored a higher microbial N
135
immobilization and a higher C availability, therefore significantly reducing soil N losses (Tapia-
136
Torres et al. 2015b). Another study in the CCB that compared two sites with different soil
137
moisture content showed that the site with the highest moisture content and concentration of
138
DOC also exhibited higher NH4+, microbial C and N concentrations, and also presented higher
139
diversity, richness and evenness of soil bacterial community compared to the dry site (López-
140
Lozano et al. 2012). Both studies suggest that differences in DOC concentration (energy
141
availability) and microbial community composition promoted different nutrient dynamics. In the
142
sites with organic matter providing lower DOC concentrations, the microbial communities may
143
be co-limited by energy and nutrients and yet they must invest more energy in order to obtain the
144
most limiting nutrients. An indicator that helps us understand how resources are reallocated by
145
the microbial community to cope with the nutrient limitation is the combination of: 1) the
146
stoichiometry ratios of C:N:P in the soil and microbial biomass (Cleveland & Liptzin 2007) and
147
2) the Threshold Elemental Ratio (TER) (Sinsabaugh & Follstad Shah 2012; Tapia-Torres et al. 148
2015a), which defines the element ratio at which growth is affected by nutrient limitation 126
(CCB), which is the study site of the present investigation, is part of the Chihuahuan desert and
127
is considered the most important wetland of Mexico for its high levels of endemism and
128
biodiversity (Souza et al. 2011). Manuscript to be reviewed Moreover, the CCB has been listed as an ultra-oligotrophic site
129
due to low P concentrations in the water and soil, which can constitute a strong potential for P
130
limitation of microbial growth (Elser et al. 2005; Tapia-Torres et al. 2015a). A study in the CCB
131
desert reported that, in the same soil type with different vegetation cover (grassland and desert
132
scrub) differences in OM content promotes variation in DOC concentration, which represents the
133
main energy source for soil microorganisms (Tapia-Torres et al. 2015b). The higher DOC
134
concentration under grassland soil compared to desert scrub soil favored a higher microbial N
135
immobilization and a higher C availability, therefore significantly reducing soil N losses (Tapia-
136
Torres et al. 2015b). Another study in the CCB that compared two sites with different soil
137
moisture content showed that the site with the highest moisture content and concentration of
138
DOC also exhibited higher NH4+, microbial C and N concentrations, and also presented higher
139
diversity, richness and evenness of soil bacterial community compared to the dry site (López-
140
Lozano et al. 2012). Both studies suggest that differences in DOC concentration (energy
141
availability) and microbial community composition promoted different nutrient dynamics. In the
142
sites with organic matter providing lower DOC concentrations, the microbial communities may
143
be co-limited by energy and nutrients and yet they must invest more energy in order to obtain the
144
most limiting nutrients. An indicator that helps us understand how resources are reallocated by
145
the microbial community to cope with the nutrient limitation is the combination of: 1) the
146
stoichiometry ratios of C:N:P in the soil and microbial biomass (Cleveland & Liptzin 2007) and
147
2) the Threshold Elemental Ratio (TER) (Sinsabaugh & Follstad Shah 2012; Tapia-Torres et al PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 172
and nutrients by the soil microbial community from two vegetation-soil systems with different
173
soil organic matter inputs in an oligotrophic desert ecosystem. Our hypothesis is that, in a site
174
with high soil resources availability, the soil microbial communities invest less energy in the
175
acquisition of nutrients (i.e. ecoenzymatic production), favoring nutrient accumulations within
176
the biomass (i.e. immobilization). Our predictions are: 1) in a site that presents low soil nutrient
177
availability (rosetophylous scrub - RS), the soil microbial community will invest more energy in
178
the production of ecoenzymes in order to depolymerize and mineralize, thus favoring nutrient
179
availability; while in a site with high soil nutrient availability (grassland - G), the soil microbial
180
community will invest more energy in biomass growth; and 2) in the site with greater soil
181
resources availability (G), the microbial community will be less vulnerable to changes in
182
precipitation. To test the hypothesis, we collected soil samples in the CCB from sites under two
183
vegetation covers (RS and G) that differ in terms of the quantity and quality of the organic matter
184
present. Collections were conducted during years: 2011(February), 2012, 2013 and 2014
185
(September), over which a noticeable variation in annual precipitation took place. The
186
ecoenzyme activity involved in the decomposition of organic matter, as well as the concentration
187
of dissolved, available and microbial biomass nutrient, were determined and compared between
188
sites and years. With the ecoenzymatic and biogeochemistry data we calculated the TERC: nutrient,
189
SEA, the nutrient ratios and performed regressions between the precipitation and the
190
concentrations and ratios of C, N and P in microbial biomass. 172
and nutrients by the soil microbial community from two vegetation-soil systems with different
173
soil organic matter inputs in an oligotrophic desert ecosystem. Our hypothesis is that, in a site
174
with high soil resources availability, the soil microbial communities invest less energy in the
175
acquisition of nutrients (i.e. ecoenzymatic production), favoring nutrient accumulations within
176
the biomass (i.e. immobilization). Manuscript to be reviewed 149
(represented by N and P, at high C:N or C:P) and by energy limitation (represented by C, at low
150
C:N or C:P) (Frost et al. 2006; Sterner & Elser 2002). The combination of stoichiometry ratios
151
and TER indicate how resources are reallocated towards enzyme activity depending on the
152
availability of energy (C) and nutrients (N and P) in the soil. This microbial co-limitation
153
between energy and nutrient acquisition was also found in CCB by comparing the TERC:N and
154
TERC:P from two sites with the same vegetation cover (grassland), but different soil moisture and
155
DOC availability values (Tapia-Torres et al. 2015a). The microbial communities were co-limited
156
by C and N in the site with higher water and C availability (Churince) and were co-limited by C
157
and P in the site with lower water and C availability (Pozas Azules). In addition, these authors
158
argue that this limitation favors an elevated allocation of N-acquisition enzymes relative to
159
energy/C enzymes in Churince, while for Pozas Azules, an elevated investment in ecoenzymes
160
of P acquisition is found (Tapia-Torres et al. 2015a). These results support the notion that soil
161
microbial communities can adjust their metabolism by allocating more resources (i.e. energy and
162
production of ecoenzymes) to the accumulation of scarcer nutrients, and fewer resources to the
163
acquisition of abundant nutrients. The ratios of C:N:P in microbial biomass are therefore
164
constrained relative to nutrient (Cleveland & Liptzin 2007) and energy availability. These studies
165
suggest that both vegetation and soil moisture content may determine differences in: 1) soil
166
nutrient dynamics, 2) the diversity of the soil microbial community and 3) the C:N:P ratios of the
167
microbial biomass in this ecosystem. 168
To date, the physiological adjustments made by the soil microbial communities under different 168
To date, the physiological adjustments made by the soil microbial communities under different
169
soil resource availability in order to obtain energy and nutrients in desert ecosystems with high
170
precipitation variability remain unclear. To elucidate this dynamic, the present study examined
171
the effects of rainfall variation on the physiological adjustments made in order to obtain energy PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed Our predictions are: 1) in a site that presents low soil nutrient
177
availability (rosetophylous scrub - RS), the soil microbial community will invest more energy in
178
the production of ecoenzymes in order to depolymerize and mineralize, thus favoring nutrient
179
availability; while in a site with high soil nutrient availability (grassland - G), the soil microbial
180
community will invest more energy in biomass growth; and 2) in the site with greater soil
181
resources availability (G), the microbial community will be less vulnerable to changes in
182
precipitation. To test the hypothesis, we collected soil samples in the CCB from sites under two
183
vegetation covers (RS and G) that differ in terms of the quantity and quality of the organic matter
184
present. Collections were conducted during years: 2011(February), 2012, 2013 and 2014
185
(September), over which a noticeable variation in annual precipitation took place. The
186
ecoenzyme activity involved in the decomposition of organic matter, as well as the concentration
187
of dissolved, available and microbial biomass nutrient, were determined and compared between
188
sites and years. With the ecoenzymatic and biogeochemistry data we calculated the TERC: nutrient,
189
SEA, the nutrient ratios and performed regressions between the precipitation and the
190
concentrations and ratios of C, N and P in microbial biomass. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 217
collection was carried out in Churince on the west side of the CCB, where Gypsisol is the
218
predominant soil type (Perroni et al. 2014b). The samples were taken from two vegetation cover
219
types, rosetophylous scrub (RS) and grassland (G), during February (2011) and September (rainy
220
of 2012, 2013 and 2014). For each vegetation cover, we sampled seven sites located at a distance
221
of 140 m apart, along a one km north-to-south transect. At each sampling site, a 4 x 4 m plot was
222
demarcated and five soil samples were taken from the first 15 cm of soil depth within the plot,
223
and mixed to produce one compound sample per site. A total of seven composite samples were
224
therefore obtained from each vegetation cover in each sampling year. The soil samples were
225
stored in black plastic bags at 4 °C until subsequent laboratory analysis. 226
227
1.3. Moisture and pH
228
Soil pH was measured in deionized water (soil/solution, 1:2 w:v) with a digital pH meter
229
(CorningTM). A subsample of 100 g was oven-dried at 75 °C to constant weight for soil moisture
230
determination using the gravimetric method. 231
232
1.4. Biogeochemical analyses
233
2.4.1. Nutrient Analysis
234
All Carbon (C) forms analyzed were determined with a Total Carbon Analyzer (UIC Mod. 235
CM5012; Chicago, USA), while nitrogen (N) and phosphorus (P) concentrations were
236
determined by colorimetric analyses, using a Bran Luebbe Auto Analyzer III (Norderstedt,
237
Germany). Microbial P and enzymatic activity were determined by colorimetric analyses using a
238
spectrophotometer Evolution 201 (Thermo Scientific Inc.). 239 217
collection was carried out in Churince on the west side of the CCB, where Gypsisol is the
218
predominant soil type (Perroni et al. 2014b). The samples were taken from two vegetation cover
219
types, rosetophylous scrub (RS) and grassland (G), during February (2011) and September (rainy
220
of 2012, 2013 and 2014). For each vegetation cover, we sampled seven sites located at a distance
221
of 140 m apart, along a one km north-to-south transect. At each sampling site, a 4 x 4 m plot was
222
demarcated and five soil samples were taken from the first 15 cm of soil depth within the plot,
223
and mixed to produce one compound sample per site. Manuscript to be reviewed 194
The study was carried out in the Cuatro Ciénegas Basin (CCB; 26°45’- 27°00’ N and 101°48’-
195
102°17’ W) in central northern Mexico, within the Chihuahuan Desert. The CCB has an area of
196
150,000 km2, with an elevation of 740 m.a.s.l. The climate is arid with an average annual
197
temperature of 21°C and 252 mm of annual rainfall, which is concentrated during the summer
198
months (http://smn.cna.gob.mx/). However in the last 30 years the annual precipitation showed a
199
high variability among years. In this study the annual precipitation was estimated as the amount
200
of rain accumulated 9-months before the sampling month. The precipitation data were obtained
201
from meteorological station 5044 “Cuatro Cienegas” located at 26°59’0’’ N and 101°04’0’’ W
202
(http://smn.cna.gob.mx/). Annual precipitation and the average temperature of the sampling
203
months varied strongly during the four studied years: The year 2011 was the wettest year (348
204
mm and 25°C), 2012 was particularly dry and hot (89 mm and 28°C) and was followed by two
205
wet years (217 mm and 230 mm for 2013 and 2014, respectively) with lower temperatures (24.9
206
and 24.8°C for 2013 and 2014, respectively). 207
Jurassic-era gypsum is the dominant parent material on the western side of the basin (McKee et
208
al. 1990). According to the WRB classification (2007), the predominant soil on the western side
209
of the basin is Gypsisol. The main vegetation types are: 1) grassland (G), dominated by
210
Sporobolus airoides (Torr.) Torr. and Allenrolfea occidentalis (S. Watson) Kuntze; 2) microphyll
211
scrub, dominated by Jatropha dioica Cerv., Larrea tridentate (DC) Cov. and Fouqueria sp
212
Kunth (Perroni et al. 2014a); and 3) rosetophylous scrub (RS) dominated by Dhasylirium
213
cedrosanum Trel., and Yucca treculeana Carriére (González 2012). 215
Mean air temperature for the sampling month (September) and annual rainfall data in each
216
studied year were obtained from the meteorological station "Rancho Pozas Azules” INIFAP. Soil 215
Mean air temperature for the sampling month (September) and annual rainfall data in each
216
studied year were obtained from the meteorological station "Rancho Pozas Azules” INIFAP. Soil PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed A total of seven composite samples were
224
therefore obtained from each vegetation cover in each sampling year. The soil samples were
225
stored in black plastic bags at 4 °C until subsequent laboratory analysis. 217
collection was carried out in Churince on the west side of the CCB, where Gypsisol is the
218
predominant soil type (Perroni et al. 2014b). The samples were taken from two vegetation cover
219
types, rosetophylous scrub (RS) and grassland (G), during February (2011) and September (rainy
220
of 2012, 2013 and 2014). For each vegetation cover, we sampled seven sites located at a distance
221
of 140 m apart, along a one km north-to-south transect. At each sampling site, a 4 x 4 m plot was
222
demarcated and five soil samples were taken from the first 15 cm of soil depth within the plot,
223
and mixed to produce one compound sample per site. A total of seven composite samples were
224
therefore obtained from each vegetation cover in each sampling year. The soil samples were
225
stored in black plastic bags at 4 °C until subsequent laboratory analysis. 234
All Carbon (C) forms analyzed were determined with a Total Carbon Analyzer (UIC Mod. 235
CM5012; Chicago, USA), while nitrogen (N) and phosphorus (P) concentrations were
236
determined by colorimetric analyses, using a Bran Luebbe Auto Analyzer III (Norderstedt,
237
Germany). Microbial P and enzymatic activity were determined by colorimetric analyses using a
238
spectrophotometer Evolution 201 (Thermo Scientific Inc.). 239 PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 240
2.4.2. Total nutrients
241
Prior to analysis of total nutrient forms, soil samples were dried and milled with a pestle and
242
agate mortar. Total C (TC) and inorganic C (IC) were determined by combustion and
243
coulometric detection (Huffman 1977). Organic total C (OTC) was calculated as the difference
244
between TC and IC. For total N (TN) and total P (TP) determination, the samples were digested
245
in a mixture of concentrated H2SO4, H2O2 (30%) and K2SO4 plus CuSO4, the latter acting as a
246
catalyst at 360 °C. Nitrogen was determined by the macro Kjeldahl method (Bremmer 1996),
247
while P was determined by the molybdate colorimetric method, following ascorbic acid
248
reduction (Murphy & Riley 1962). 249
250
2.4.3. Dissolved and available nutrients and those within the microbial biomass
251
The dissolved, available and microbial nutrient forms were extracted from fresh field
252
samples. Dissolved nutrients were extracted from 20 g of soil with deionized water a
253
for 45 min and then filtering through a Whatman No. 42 and a 0.45 μm nitrocellulos
254
(Jones & Willett 2006). The filtrate was used to determine the total dissolved C (TD 249
250
2.4.3. Dissolved and available nutrients and those within the microbial biomass
251
The dissolved, available and microbial nutrient forms were extracted from fresh field soil
252
samples. Dissolved nutrients were extracted from 20 g of soil with deionized water after shaking
253
for 45 min and then filtering through a Whatman No. 42 and a 0.45 μm nitrocellulose membrane
254
(Jones & Willett 2006). The filtrate was used to determine the total dissolved C (TDC), as
255
measured with an Auto Analyzer of carbon (TOC CM 5012) module for liquids (UIC-
256
COULOMETRICS). Inorganic dissolved C (IDC) was determined in an acidification module
257
CM5130. One aliquot of the filtrate was used to determine ammonium (DNH4+) and dissolved
258
inorganic P (DIP) in a deionized water extract. Total dissolved N and P (TDN and TDP,
259
respectively) were digested in a mixture of concentrated H2SO4, H2O2 (30%) at 250 °C. Nitrogen
260
was determined by the macro Kjeldahl method (Bremmer 1996), while P was determined by the
261
molybdate colorimetric method, following ascorbic acid reduction (Murphy & Riley 1962). 250
2.4.3. Manuscript to be reviewed Dissolved and available nutrients and those within the microbial biomass
251
The dissolved, available and microbial nutrient forms were extracted from fresh field soil
252
samples. Dissolved nutrients were extracted from 20 g of soil with deionized water after shaking
253
for 45 min and then filtering through a Whatman No. 42 and a 0.45 μm nitrocellulose membrane
254
(Jones & Willett 2006). The filtrate was used to determine the total dissolved C (TDC), as
255
measured with an Auto Analyzer of carbon (TOC CM 5012) module for liquids (UIC-
256
COULOMETRICS). Inorganic dissolved C (IDC) was determined in an acidification module
257
CM5130. One aliquot of the filtrate was used to determine ammonium (DNH4+) and dissolved
258
inorganic P (DIP) in a deionized water extract. Total dissolved N and P (TDN and TDP,
259
respectively) were digested in a mixture of concentrated H SO
H O (30%) at 250 °C Nitrogen PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 262
Dissolved organic C, N and P (DOC, DON and DOP respectively) values were calculated as the
263
difference between the total dissolved forms and the inorganic dissolved forms. 264
Available inorganic nitrogen forms (NH4+ and NO3-) were extracted from 10 g of soil with 2M
265
KCl, followed by filtration through a Whatman No. 1 paper filter, and determined
266
colorimetrically by the phenol-hypochlorite method (Technicon 1977). Available inorganic
267
phosphorous (Pi) was extracted with 0.5 M NaHCO3, pH 8.5 (Tiessen & Moir 2008) and
268
determined colorimetrically using the molybdate-ascorbic acid method (Murphy & Riley 1962). 269
Carbon (Cmic) and N (Nmic) concentrations within the microbial biomass were determined from
270
20 g of soil by the chloroform fumigation extraction method (Vance et al. 1987). Fumigated and
271
non-fumigated samples were incubated for 24 h at 25 °C and constant relative humidity. Cmic
272
and Nmic were extracted from fumigated and non-fumigated samples with 0.5 MK2SO4, filtered
273
through a 0.45 μm nitrocellulose membrane (Brookes et al. 1984). Carbon concentration was
274
measured from each extract, as the total (TC) and inorganic (IC) carbon contents, using the
275
method described before. The difference between TC and IC was used for Cmic calculation. To
276
determine the Nmic concentration one aliquot of the filtrate extracted was acid digested and
277
determined as TN by Macro-Kjeldahl method (Brookes et al. 1984). Phosphorus within
278
microbial biomass (Pmic) was extracted from 5 g of soil by the chloroform fumigation extraction
279
and incubation method (Vance et al. 1987). Pmic was extracted using NaCO3 0.5M, pH 8.5 and
280
digested in a mixture of H2SO4 11N and (NH4)2S2O8 at 50%, with the latter acting as a catalyst at
281
120 °C (Lajtha et al. 1999). Pmic was determined colorimetrically by the molybdate-ascorbic
282
acid method (Murphy & Riley 1962). The values of Cmic, Nmic and Pmic were calculated as
283
the difference between fumigated and non-fumigated samples using correction factors of KEC
284
0.45 (Joergensen 1996), KEN 0.54 (Joergensen & Mueller 1996) and KP 0.4 (Lajtha et al. 1999) 262
Dissolved organic C, N and P (DOC, DON and DOP respectively) values were calculated as the
263
difference between the total dissolved forms and the inorganic dissolved forms. Manuscript to be reviewed 264
Available inorganic nitrogen forms (NH4+ and NO3-) were extracted from 10 g of soil with 2M
265
KCl, followed by filtration through a Whatman No. 1 paper filter, and determined
266
colorimetrically by the phenol-hypochlorite method (Technicon 1977). Available inorganic
267
phosphorous (Pi) was extracted with 0.5 M NaHCO3, pH 8.5 (Tiessen & Moir 2008) and
268
determined colorimetrically using the molybdate-ascorbic acid method (Murphy & Riley 1962). 269
Carbon (Cmic) and N (Nmic) concentrations within the microbial biomass were determined from
270
20 g of soil by the chloroform fumigation extraction method (Vance et al. 1987). Fumigated and
271
non-fumigated samples were incubated for 24 h at 25 °C and constant relative humidity. Cmic
272
and Nmic were extracted from fumigated and non-fumigated samples with 0.5 MK2SO4, filtered
273
through a 0.45 μm nitrocellulose membrane (Brookes et al. 1984). Carbon concentration was
274
measured from each extract, as the total (TC) and inorganic (IC) carbon contents, using the
275
method described before. The difference between TC and IC was used for Cmic calculation. To
276
determine the Nmic concentration one aliquot of the filtrate extracted was acid digested and
277
determined as TN by Macro-Kjeldahl method (Brookes et al. 1984). Phosphorus within
278
microbial biomass (Pmic) was extracted from 5 g of soil by the chloroform fumigation extraction
279
and incubation method (Vance et al. 1987). Pmic was extracted using NaCO3 0.5M, pH 8.5 and
280
digested in a mixture of H2SO4 11N and (NH4)2S2O8 at 50%, with the latter acting as a catalyst at
281
120 °C (Lajtha et al. 1999). Pmic was determined colorimetrically by the molybdate-ascorbic
282
acid method (Murphy & Riley 1962). The values of Cmic, Nmic and Pmic were calculated as
283
the difference between fumigated and non-fumigated samples using correction factors of KEC 262
Dissolved organic C, N and P (DOC, DON and DOP respectively) values were calculated as the
263
difference between the total dissolved forms and the inorganic dissolved forms. 262
Dissolved organic C, N and P (DOC, DON and DOP respectively) values were calculated as the
263
difference between the total dissolved forms and the inorganic dissolved forms. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 285
for Cmic, Nmic and Pmic, respectively. Finally, the values of Cmic, Nmic and Pmic were
286
corrected to a dry soil basis. 287
288
1.5. Molecular analysis
289
Bacterial composition analysis was performed on the samples from the wettest year (2011). We
290
extracted DNA from each soil sample using the methodology described in López-Lozano (2013)
291
and sent it to J. Craig Venter Institute (JCVI) in order to construct a 16S library using 454
292
ROCHE tag, 50,000 reads per site of 500 bp and primers 341F-926R. Sequences were trimmed
293
and chimeras eliminated using JCVI protocols. Taxa were assigned using Blast via JCVI
294
pipeline, these methods are detailed by Tanenbaum et al. (2010). 295
296
1.6. Ecoenzyme Activity Analyses
297
The activities of six ecoenzymes (extracellular enzymes) involved in the cleavage of organic
298
molecules with C, N and P were measured: β-1, 4-glucosidase (BG), cellobiohydrolase (CBH),
299
β-1, 4-N-acetylglucosaminidase (NAG), polyphenol oxidase (PPO), phosphomonoesterase
300
(PME) and phosphodiesterase (PDE), using assay techniques reported by Tabatabai & Bremner
301
(1969), Eivazi & Tabatabai (1977), Eivazi & Tabatabai (1988),Verchot & Borelli (2005) and
302
Johannes & Majcherczyk (2000). 303
For all ecoenzymes, we used 2 g of fresh soil and 30 ml of modified universal buffer (MUB) at
304
pH 9 for ecoenzyme extraction. Three replicates and two control samples (soil extract with no
305
b t t
d
MUB
ith
b t t )
i
l d d
All 285
for Cmic, Nmic and Pmic, respectively. Finally, the values of Cmic, Nmic and Pmic were
286
corrected to a dry soil basis. 296
1.6. Ecoenzyme Activity Analyses
297
The activities of six ecoenzymes (extracellular enzymes) involved in the cleavage of organic
298
molecules with C, N and P were measured: β-1, 4-glucosidase (BG), cellobiohydrolase (CBH),
299
β-1, 4-N-acetylglucosaminidase (NAG), polyphenol oxidase (PPO), phosphomonoesterase
300
(PME) and phosphodiesterase (PDE), using assay techniques reported by Tabatabai & Bremner
301
(1969), Eivazi & Tabatabai (1977), Eivazi & Tabatabai (1988),Verchot & Borelli (2005) and
302
Johannes & Majcherczyk (2000). Manuscript to be reviewed 297
The activities of six ecoenzymes (extracellular enzymes) involved in the cleavage of organic
298
molecules with C, N and P were measured: β-1, 4-glucosidase (BG), cellobiohydrolase (CBH),
299
β-1, 4-N-acetylglucosaminidase (NAG), polyphenol oxidase (PPO), phosphomonoesterase
300
(PME) and phosphodiesterase (PDE), using assay techniques reported by Tabatabai & Bremner
301
(1969), Eivazi & Tabatabai (1977), Eivazi & Tabatabai (1988),Verchot & Borelli (2005) and
302
Johannes & Majcherczyk (2000). 303
For all ecoenzymes, we used 2 g of fresh soil and 30 ml of modified universal buffer (MUB) at
304
pH 9 for ecoenzyme extraction. Three replicates and two control samples (soil extract with no
305
substrate, and pure MUB with substrate) were included per assay. All ecoenzyme assays were
306
incubated at 40°C: the BG and CBH for 2h, NAG for 3h, PPO for 2.5h, PME and PDE 1.25h. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 307
Following the incubation period, the tubes were centrifuged at 10,000 rpm for 2 minutes and
308
750μl of supernatant was recovered. 307
Following the incubation period, the tubes were centrifuged at 10,000 rpm for 2 minutes and
308
750μl of supernatant was recovered. 307
Following the incubation period, the tubes were centrifuged at 10,000 rpm for 2 minutes and
308
750μl of supernatant was recovered. 309
For all ecoenzymes with substrates containing p-nitrophenol (pNP), we diluted the supernatant in
310
2 ml of deionized water with 75 μl of NaOH and measured the absorbance of pNP liberated at
311
410 nm on an Evolution 201 spectrophotometer (Thermo Scientific Inc.). For the PPO, we used
312
2,2'-Azinobis [3-ethylbenzothiazoline-6-sulfonic acid]-diammonium salt (ABTS) as a substrate. 313
The resulting supernatant was measured directly at 410 nm. Ecoenzyme activities were
314
expressed as nanomoles of pNP per gram of dry soil per hour (nmol pNP [g SDE]-1 h-1) for
315
substrates containing p-nitrophenol (pNP) and O2 formed per gram of dry soil per hour (nmolO2
316
[g SDE]-1 h-1) for the PPO, respectively. Specific enzymatic activity was calculated using
317
equations (1) to (3) (Chavez-Vergara et al. 2014; Waldrop et al. 2000): 318
(1)
𝑆𝐸𝐴 µ𝑚𝑜𝑙/(𝑚𝑔𝐶𝑚𝑖𝑐 ℎ ) = 𝐴/(𝐶𝑚𝑖𝑐 𝑥 0.001) 319
𝑆𝐸𝐴 µ𝑚𝑜𝑙/(𝑚𝑔𝑁𝑚𝑖𝑐 ℎ) = 𝐵/(𝑁𝑚𝑖𝑐 𝑥 0.001) 320
(3)
𝑆𝐸𝐴 µ𝑚𝑜𝑙/(𝑚𝑔𝑃𝑚𝑖𝑐 ℎ) = 𝐶/(𝑃𝑚𝑖𝑐 𝑥 0.001) (3) 321
Where A is the enzymatic activity of BG or CBH or PPO, B is the enzymatic activity of NAG 321
Where A is the enzymatic activity of BG or CBH or PPO, B is the enzymatic activity of NAG 322
and C is the enzymatic activity of PME or PDE. 325
1.7.1. Biogeochemistry and ecoenzymatic analysis 326
Soil biogeochemistry and ecoenzymatic data were subjected to a repeated measures analysis of
327
variance (RMANOVA) (Von Ende 2001). Vegetation cover types (RS and G) were considered
328
as a between-subject factor and year (2012, 2013 and 2014), and their interaction, were 329
considered as within-subject factors. When RMANOVA indicated significant factor effects, PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 330
mean comparisons were performed with Tukey's multiple comparisons test (Von Ende 2001). 330
mean comparisons were performed with Tukey's multiple comparisons test (Von Ende 2001). 330
mean comparisons were performed with Tukey's multiple comparisons test (Von Ende 2001). 331
Ecoenzyme activities were normalized to units per g of available organic carbon (OC) using the
332
DOC data corresponding to each sample (Tapia-Torres et al. 2015a). Data were loge-transformed
333
prior to regression analysis in order to conform to the conventions of stoichiometric analyses and
334
to normalize variance (Sinsabaugh & Follstad Shah 2012; Sterner & Elser 2002). After that,
335
relationships between ecoenzyme activities were calculated with a type II regression, using
336
SMATR (SMATR 2007). 330
mean comparisons were performed with Tukey's multiple comparisons test (Von Ende 2001). 331
Ecoenzyme activities were normalized to units per g of available organic carbon (OC) using the
332
DOC data corresponding to each sample (Tapia-Torres et al. 2015a). Data were loge-transformed
333
prior to regression analysis in order to conform to the conventions of stoichiometric analyses and
334
to normalize variance (Sinsabaugh & Follstad Shah 2012; Sterner & Elser 2002). After that,
335
relationships between ecoenzyme activities were calculated with a type II regression, using
336
SMATR (SMATR 2007). 337
To detect the relationship between precipitation and nutrients immobilized by microbial biomass,
338
we applied two simple regression analyses using the annual accumulated precipitation prior to
339
the sampling date with: 1) nutrient concentration within the microbial biomass (Cmic, Nmic and
340
Pmic) and 2) the microbial biomass nutrient ratios (Cmic:Nmic, Cmic:Pmic and Nmic:Pmic). 341
The data used in the regression analyses corresponded to the years 2011, 2012, 2013 and 2014. 342
2.7.2. Stoichiometric analyses and threshold elemental ratio 337
To detect the relationship between precipitation and nutrients immobilized by microbial biomass,
338
we applied two simple regression analyses using the annual accumulated precipitation prior to
339
the sampling date with: 1) nutrient concentration within the microbial biomass (Cmic, Nmic and
340
Pmic) and 2) the microbial biomass nutrient ratios (Cmic:Nmic, Cmic:Pmic and Nmic:Pmic). 341
The data used in the regression analyses corresponded to the years 2011, 2012, 2013 and 2014. 342
2.7.2. Manuscript to be reviewed Stoichiometric analyses and threshold elemental ratio 343
We calculated the degree of soil community-level microbial C:N and C:P homeostasis by
344
calculating the slope of loge C:NR (resources) versus loge C:NB (microbial biomass) or the slope
345
of loge C:PR versus loge C:PB scatterplot (Sterner & Elser 2002). Moreover, we followed
346
Sinsabaugh et al. (2009) in order to calculate the TER for C:N and C:P to relate the measured
347
ecoenzyme activity with Ecological Stoichiometry Theory (EST) and the Metabolic Theory of
348
Ecology (MTE), using equations (5) and (6): 349
TERC:N= ((BG/NAG)BC:N)/n0 (5) 351
Where TERC:N and TERC:P are the threshold ratios (dimensionless), BG/NAG is the
352
ecoenzymatic activity ratio for -1,4-glucosidase and -1,4-N-acetylglucosaminidase, BG/PME PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 353
is the ecoenzymatic ratio for -1,4-glucosidase and phosphomonoesterase, BC:N and BC:P are
354
the C:N or C:P ratios of the microbial biomass (respectively) and n0 and p0 are the
355
dimensionless normalization constants for N and P, respectively. These normalization constants
356
p0 and n0 are the intercepts in the SMA regressions for loge (BG) vs. loge (NAG) and loge (BG)
357
vs. loge (PME) respectively (Tapia-Torres et al. 2015a). For a more detailed analysis of the
358
derivation of the equations, see Sinsabaugh et al. (2009). 359
360
2. Results
361
2.1. Soil moisture, and pH
362
Regardless of vegetation cover, soil moisture was higher in 2013 and 2014 than in 2012; while
363
the G soil had higher soil moisture than the RS soil, regardless of year (Table 1 and 2). In the
364
driest year (2012), soil pH was higher than in the wetter years (2013 and 2014), with an
365
exception in the G soil in 2014 (Table 1 and 2). Soil pH correlated with annual precipitation in
366
both sites (R2 = -0.85 and R2 = -0.61 for RS and G, respectively), as well as soil moisture
367
correlated with annual precipitation (R2 = 0.76 and R2 = 0.88 for RS and G, respectively). 368
369
2.2. Dissolved organic nutrients and available nutrients
370
For the two vegetation covers, the lowest values of DOC, DON and DOP were found in 2012
371
(Table 1). In this year, the RS and the G soils had similar DOC and DOP concentrations, while
h G
il h d
hi h
DON
i
h
h
b
il
h G
il h d hi h 353
is the ecoenzymatic ratio for -1,4-glucosidase and phosphomonoesterase, BC:N and BC:P are
354
the C:N or C:P ratios of the microbial biomass (respectively) and n0 and p0 are the
355
dimensionless normalization constants for N and P, respectively. These normalization constants
356
p0 and n0 are the intercepts in the SMA regressions for loge (BG) vs. loge (NAG) and loge (BG)
357
vs. loge (PME) respectively (Tapia-Torres et al. 2015a). For a more detailed analysis of the
358
derivation of the equations, see Sinsabaugh et al. (2009). Manuscript to be reviewed 353
is the ecoenzymatic ratio for -1,4-glucosidase and phosphomonoesterase, BC:N and BC:P are
354
the C:N or C:P ratios of the microbial biomass (respectively) and n0 and p0 are the
355
dimensionless normalization constants for N and P, respectively. These normalization constants
356
p0 and n0 are the intercepts in the SMA regressions for loge (BG) vs. loge (NAG) and loge (BG)
357
vs. loge (PME) respectively (Tapia-Torres et al. 2015a). For a more detailed analysis of the
358
derivation of the equations, see Sinsabaugh et al. (2009). 369
2.2. Dissolved organic nutrients and available nutrients 370
For the two vegetation covers, the lowest values of DOC, DON and DOP were found in 2012
371
(Table 1). In this year, the RS and the G soils had similar DOC and DOP concentrations, while
372
the G soil had a higher DON concentration than the scrub soil; moreover, the G soil had higher
373
dissolved organic nutrient concentrations than in the RS soil in both 2013 and 2014 (Table 1 and
374
2). Consequently, the DOC:DON ratio was lower in 2012 than in the other two years (2013 and 370
For the two vegetation covers, the lowest values of DOC, DON and DOP were found in 2012
371
(Table 1). In this year, the RS and the G soils had similar DOC and DOP concentrations, while
372
the G soil had a higher DON concentration than the scrub soil; moreover, the G soil had higher
373
dissolved organic nutrient concentrations than in the RS soil in both 2013 and 2014 (Table 1 and
374
2). Consequently, the DOC:DON ratio was lower in 2012 than in the other two years (2013 and PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 375
2014) and the RS soil had lower values than the G soils (3.5 and 4.9, respectively); the RS soil
376
also had lower DOC:DOP ratios than the G soil (9 and 16, respectively). 377
The year trends of available NH4+ concentration differed between the two vegetation cover types. 378
Available NH4+ concentration was similar over the three years in the RS soil, while G soil
379
samples from 2012 and 2013 had the lowest and the highest NH4+ concentrations, respectively
380
(Table 1 and 2). However, the G soil had higher values than the RS soil in the three studied
381
years. In contrast, the NO3- concentration was higher in the samples collected in 2012 than those
382
of the other two years, while the RS soil had higher NO3- concentration than the G soil only in
383
the 2012 samples (Table 1 and 2). The 2012 samples had lower available P concentration than in
384
those collected in the other two years, and the G samples had 40% higher available P
385
concentration than the RS samples, regardless of the sampling year (4.1 and 2.9 µg P g-1,
386
respectively). 387
388
2.3. Microbial nutrients and ecoenzymatic activities
389
The highest and the lowest values of Cmic and Nmic were found in 2014 and 2012, respectively,
390
and Cmic values of the G soil samples were 39% higher than in the RS soil samples, regardless
391
of sampling year (254 and 184 µg C g-1, respectively). This was also the case with the Nmic and
392
Pmic concentrations, with an exception in the 2012 samples (Table 3 and 4). In contrast, Pmic
393
concentrations presented no differences among years within the RS samples, while the 2012
394
samples had lower Pmic values than was the case in the other two years, within the G samples
395
(Table 3 and 4). The 2014 samples had lower Cmic:Nmic than the other two years regardless of
396
vegetation cover type (2012 and 2013), while the lowest and the highest Cmic:Pmic and
397
Nmic:Pmic ratios were found in 2012 and 2014, respectively (Table 3 and 4). The RS soil
Manuscript to be review 375
2014) and the RS soil had lower values than the G soils (3.5 and 4.9, respectively); the RS soil
376
also had lower DOC:DOP ratios than the G soil (9 and 16, respectively). Manuscript to be reviewed 375
2014) and the RS soil had lower values than the G soils (3.5 and 4.9, respectively); the RS soil
376
also had lower DOC:DOP ratios than the G soil (9 and 16, respectively). 375
2014) and the RS soil had lower values than the G soils (3.5 and 4.9, respectively); the RS soil
376
also had lower DOC:DOP ratios than the G soil (9 and 16, respectively). 377
The year trends of available NH4+ concentration differed between the two vegetation cover types. 378
Available NH4+ concentration was similar over the three years in the RS soil, while G soil
379
samples from 2012 and 2013 had the lowest and the highest NH4+ concentrations, respectively
380
(Table 1 and 2). However, the G soil had higher values than the RS soil in the three studied
381
years. In contrast, the NO3- concentration was higher in the samples collected in 2012 than those
382
of the other two years, while the RS soil had higher NO3- concentration than the G soil only in
383
the 2012 samples (Table 1 and 2). The 2012 samples had lower available P concentration than in
384
those collected in the other two years, and the G samples had 40% higher available P
385
concentration than the RS samples, regardless of the sampling year (4.1 and 2.9 µg P g-1,
386
respectively). 388
2.3. Microbial nutrients and ecoenzymatic activities y
389
The highest and the lowest values of Cmic and Nmic were found in 2014 and 2012, respectively,
390
and Cmic values of the G soil samples were 39% higher than in the RS soil samples, regardless
391
of sampling year (254 and 184 µg C g-1, respectively). This was also the case with the Nmic and
392
Pmic concentrations, with an exception in the 2012 samples (Table 3 and 4). In contrast, Pmic
393
concentrations presented no differences among years within the RS samples, while the 2012
394
samples had lower Pmic values than was the case in the other two years, within the G samples
395
(Table 3 and 4). The 2014 samples had lower Cmic:Nmic than the other two years regardless of
396
vegetation cover type (2012 and 2013), while the lowest and the highest Cmic:Pmic and
397
Nmic:Pmic ratios were found in 2012 and 2014, respectively (Table 3 and 4). The RS soil PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 421
number of unclassified bacteria; 26% for the RS soil and 20% for the G soil (Fig. 4). In the two
422
vegetation cover types, the Proteobacteria was the most abundant bacterial phylum, accounting
423
for 20% in the RS soil and 30% in the G soil. Similarly, Actinobacteria was the second most
424
dominant phylum in the RS soil and in the G soil, with an abundance of 14% in both soils. 425
Interestingly, the Cyanobacteria was the third most dominant phylum, with 13% of abundance
426
both soils, suggesting the importance of the desert crust in both sites. Other important phyla
427
observed were: Chloroflexi (10%), Bacteroidetes (5%), Plantomycetes (4%), Firmicutes (4%),
428
Nitrospira (1% in the RS and 0.5% in the G soils) and Acidobacteria (6% in RS and 0.8% in G;
429
Fig. 4). 430
431
3.5 Ecoenzymatic stoichiometry, homeostasis and threshold elemental ratios
432
In all of the model II regressions analyzed, there were no differences found in slopes between
433
soils of the two vegetation cover types within sampling years (Figures S1 and S2). To test the
434
strength of stoichiometric homeostasis, we analyzed for associations between microbial biomas
435
elemental ratios and those in the soil resources (Tapia-Torres et al. 2015a). In both soil
436
vegetation cover types, the relationships between log C:NR and log C:NB, and between log C:PR
437
and log C:PB did not differ from zero (p > 0.05), regardless of year (Figures S1 and S2);
438
indicating strong community-level elemental homeostasis in the soil of both sites. 439
Moreover, we used the parameters generated from the type II regressions using enzymatic data
440
and microbial C:N:P stoichiometric values to estimate TERC:N and TERC:P values. The lowest
441
TERC:N values were observed in 2014 (wet year), but no differences were observed between 20
442
and 2013, or even between vegetation cover types (RS and G) among study years (Fig. 5). The
443
opposite was found for TERC:P, where we obtained the lowest value in the dry year (2012), but 421
number of unclassified bacteria; 26% for the RS soil and 20% for the G soil (Fig. 4). In the two
422
vegetation cover types, the Proteobacteria was the most abundant bacterial phylum, accounting
423
for 20% in the RS soil and 30% in the G soil. Manuscript to be reviewed 398
samples had higher Cmic:Pmic and Nmic:Pmic ratios than in the G soil samples, with an
399
exception in the 2012 samples (Table 3 and 4). 398
samples had higher Cmic:Pmic and Nmic:Pmic ratios than in the G soil samples, with an
399
exception in the 2012 samples (Table 3 and 4). 398
samples had higher Cmic:Pmic and Nmic:Pmic ratios than in the G soil samples, with an
399
exception in the 2012 samples (Table 3 and 4). 400
Significant positive correlations were observed between precipitation and immobilized nutrients
401
within the microbial biomass (Cmic, Nmic, Pmic), in both soils. Moreover, significant positive
402
correlations were detected between precipitation and the Cmic:Nmic and Cmic:Pmic ratios in the
403
RS soil and the Cmic:Pmic and Nmic:Pmic ratios in the G soil. The slopes of the regression with
404
Cmic and Nmic were higher in the G soil and lower in the RS soil (Fig.1, Fig. 2 and Table S1). 405
The specificenzymatic activity of BG under both vegetation cover types was lower in the wet
406
(2014) than in the dry year (2012; Figure 3a, Table 4), while that of CBH in the dry year was
407
lower than in both wet years (2013 and 2014), in both vegetation covers (Fig. 3b). The specific
408
enzymatic activity of the PPO in the scrub soil did not differ among years, while the dry year
409
(2012) had lower values than the wet years (2013 and 2014) in the G soil (Fig. 3c and Table 4). 410
Furthermore, the G soil had higher specific PPO enzimatic activity than the RS soil in the wet
411
year (2014). In contrast, the wet year (2014) had the lowest NAG specific enzymatic activity
412
under both vegetation cover types, and the RS soil had lower values only in the dry year (2012;
413
Fig. 3d). The specific enzymatic activity of PME and PDE was similar, and the lowest values of
414
specific enzymatic activity were in the driest year (2012). In the two wet years, the RS soil
415
presented higher specific activities than the G soil (Fig 3e and 3f). PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed Similarly, Actinobacteria was the second most
424
dominant phylum in the RS soil and in the G soil, with an abundance of 14% in both soils. 438
indicating strong community-level elemental homeostasis in the soil of both sites. 439
Moreover, we used the parameters generated from the type II regressions using enzymatic data
440
and microbial C:N:P stoichiometric values to estimate TERC:N and TERC:P values. The lowest
441
TERC:N values were observed in 2014 (wet year), but no differences were observed between 2012
442
and 2013, or even between vegetation cover types (RS and G) among study years (Fig. 5). The
443
opposite was found for TERC:P, where we obtained the lowest value in the dry year (2012), but 439
Moreover, we used the parameters generated from the type II regressions using enzymatic data
440
and microbial C:N:P stoichiometric values to estimate TERC:N and TERC:P values. The lowest
441
TERC:N values were observed in 2014 (wet year), but no differences were observed between 2012
442
and 2013, or even between vegetation cover types (RS and G) among study years (Fig. 5). The
443
opposite was found for TERC:P, where we obtained the lowest value in the dry year (2012), but PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 444
only in the RS soil. For the dry year (2012), no differences were observed between vegetation
445
cover types, while we observed lower TERC:P values in the G soil than in the RS soil for the wet
446
years (2013 and 2014; Fig. 5). 447
448
3. Discussion
449
3.1. Reallocation of resources by the soil microbial community
450
Our first prediction, that the soil microbial community invests more energy in the production of
451
ecoenzymes to acquire nutrients in sites of low resource availability, such as the RS soil, was
452
confirmed. We observed that the RS soil showed a lower concentration of available P than the G
453
soil in the three years studied and, consequently, the RS soil microbial community invested
454
more energy in the acquisition of P (increased enzymatic activity of phosphomonoesterase and
455
phosphodiesterase) than the G soil microbial community only during the two wet years. In
456
contrast, the G soil had higher Cmic, Nmic and Pmic concentrations and lower enzymatic
457
activity of phosphomonoesterase and phosphodiesterase than the RS soil during both wet years,
458
which also supports our prediction (Fig 1 and Table 3). These results suggest that the microbial
459
community in the RS soil, with lower resource availability, must reduce growth as a result of: 1) the
460
physiological cost associated with a low reallocation to P-rich ribosomal RNA, as suggested by
461
the growth rate hypothesis (GRH) (Sinsabaugh & Follstad Shah 2012; Sterner & Elser 2002;
462
Zechmeister-Boltenstern et al. 2015) and 2) the required investment of energy towards the
463
acquisition of P in order to produce ecoenzymes (Evans & Wallenstein 2012; Schimel et al. 2007;
464
Wallenstein & Hall 2012). The microbial C:N:P ratio was greater in the RS soil (127:19:1) than in 444
only in the RS soil. For the dry year (2012), no differences were observed between vegetation
445
cover types, while we observed lower TERC:P values in the G soil than in the RS soil for the wet
446
years (2013 and 2014; Fig. 5). 444
only in the RS soil. For the dry year (2012),
445
cover types, while we observed lower TERC
446
years (2013 and 2014; Fig. 5). 447
448
3. Discussion
449
3.1. Manuscript to be reviewed Reallocation of resources by the soil m
450
Our first prediction, that the soil microbial c
451
ecoenzymes to acquire nutrients in sites of l
452
confirmed. We observed that the RS soil sh
453
soil in the three years studied and, conseque
454
more energy in the acquisition of P (increas
455
phosphodiesterase) than the G soil microbia
456
contrast, the G soil had higher Cmic, Nmic
457
activity of phosphomonoesterase and phosp
458
which also supports our prediction (Fig 1 an
459
community in the RS soil, with lower resource a
460
physiological cost associated with a low rea
461
the growth rate hypothesis (GRH) (Sinsabau
462
Zechmeister-Boltenstern et al. 2015) and 2)
463
acquisition of P in order to produce ecoenzy
464
Wallenstein & Hall 2012) The microbial C:N: 449
3.1. Reallocation of resources by the soil microbial community 449
3.1. Reallocation of resources by the soil microbial community
450
Our first prediction, that the soil microbial community invests more energy in the production of
451
ecoenzymes to acquire nutrients in sites of low resource availability, such as the RS soil, was
452
confirmed. We observed that the RS soil showed a lower concentration of available P than the G
453
soil in the three years studied and, consequently, the RS soil microbial community invested
454
more energy in the acquisition of P (increased enzymatic activity of phosphomonoesterase and
455
phosphodiesterase) than the G soil microbial community only during the two wet years. In
456
contrast, the G soil had higher Cmic, Nmic and Pmic concentrations and lower enzymatic
457
activity of phosphomonoesterase and phosphodiesterase than the RS soil during both wet years,
458
which also supports our prediction (Fig 1 and Table 3). These results suggest that the microbial
459
community in the RS soil, with lower resource availability, must reduce growth as a result of: 1) the
460
physiological cost associated with a low reallocation to P-rich ribosomal RNA, as suggested by
461
the growth rate hypothesis (GRH) (Sinsabaugh & Follstad Shah 2012; Sterner & Elser 2002;
462
Zechmeister-Boltenstern et al. 2015) and 2) the required investment of energy towards the
463
acquisition of P in order to produce ecoenzymes (Evans & Wallenstein 2012; Schimel et al. 2007;
464
Wallenstein & Hall 2012). Manuscript to be reviewed 467
and a high capacity for P occlusion within inorganic molecules, mainly by Ca-bound (Perroni et
468
al. 2014b). Therefore, the main source of available P is mineralization of organic P mediated by
469
phosphatase activity (Waring et al. 2014). Among organic P molecules, phosphodiester forms are
470
the preferred substrate in P-limited ecosystems (Karl 2014; Tapia-Torres et al. 2016), although
471
phosphomonoester forms may also be an important source of available P in most soils (Turner et
472
al. 2003). In our study sites, phosphodiesterase activity was almost ten times higher than that of
473
phosphomonoesterase, mainly in the scrub soil, suggesting mineralization of phosphodiesters as
474
the main source of soil available P. Several bacteria isolates from CCB soils prefer to grow in
475
DNA as a P source, associated with phosphodiesterase activity (Tapia-Torres et al. 2016). We
476
suggest that the main P source in sites with low nutrient availability, such as the RS soil, is
477
recycling of the organic molecules that are the product of cellular lysis. 478
However, the G soil had higher enzymatic activity of polyphenol oxidase in the wet year 2014
479
than was the case in the RS soil. This result is consistent with other studies (Sinsabaugh 2010;
480
Sinsabaugh & Follstad Shah 2011), which have reported that polyphenol oxidase activity does
481
not present the same behavior as the β-1, 4-glucosidase and other hydrolases that degrade labile
482
C. Microbial community size begins to be limited by the availability of labile C, which produces
483
a change in the microbial community composition towards microbial guilds with lower growth
484
rates (low concentration of Cmic), but with the capacity to produce polyphenol oxidase to break
485
down structurally complex molecules and obtain C (Moorhead & Sinsabaugh 2006). This
486
situation is comparable to the conditions of the G soil in the wet year 2014, where the microbial
487
community was required to cleave lignin in order to maintain its growth rate. 488
Furthermore, the differences in soil nutrient dynamics between both sites can be strongly
489
affected by soil microbial composition. While analyses of soil bacterial composition were only 467
and a high capacity for P occlusion within inorganic molecules, mainly by Ca-bound (Perroni et
468
al. 2014b). Therefore, the main source of available P is mineralization of organic P mediated by
469
phosphatase activity (Waring et al. 2014). Manuscript to be reviewed Among organic P molecules, phosphodiester forms are
470
the preferred substrate in P-limited ecosystems (Karl 2014; Tapia-Torres et al. 2016), although
471
phosphomonoester forms may also be an important source of available P in most soils (Turner et
472
al. 2003). In our study sites, phosphodiesterase activity was almost ten times higher than that of
473
phosphomonoesterase, mainly in the scrub soil, suggesting mineralization of phosphodiesters as
474
the main source of soil available P. Several bacteria isolates from CCB soils prefer to grow in
475
DNA as a P source, associated with phosphodiesterase activity (Tapia-Torres et al. 2016). We
476
suggest that the main P source in sites with low nutrient availability, such as the RS soil, is
477
recycling of the organic molecules that are the product of cellular lysis. 478
However, the G soil had higher enzymatic activity of polyphenol oxidase in the wet year 2014
479
than was the case in the RS soil. This result is consistent with other studies (Sinsabaugh 2010;
480
Sinsabaugh & Follstad Shah 2011), which have reported that polyphenol oxidase activity does
481
not present the same behavior as the β-1, 4-glucosidase and other hydrolases that degrade labile
482
C. Microbial community size begins to be limited by the availability of labile C, which produces
483
a change in the microbial community composition towards microbial guilds with lower growth
484
rates (low concentration of Cmic), but with the capacity to produce polyphenol oxidase to break
485
down structurally complex molecules and obtain C (Moorhead & Sinsabaugh 2006). This
486
situation is comparable to the conditions of the G soil in the wet year 2014, where the microbial
487
community was required to cleave lignin in order to maintain its growth rate. 478
However, the G soil had higher enzymatic activity of polyphenol oxidase in the wet year 2014
479
than was the case in the RS soil. This result is consistent with other studies (Sinsabaugh 2010;
480
Sinsabaugh & Follstad Shah 2011), which have reported that polyphenol oxidase activity does
481
not present the same behavior as the β-1, 4-glucosidase and other hydrolases that degrade labile
482
C. Manuscript to be reviewed The microbial C:N:P ratio was greater in the RS soil (127:19:1) than in
465
the G soil (63:10:1), suggesting that the microbial community in the former site is more P-
466
constrained (Cleveland & Liptzin 2007). The studied soils are characterized by low P availability 450
Our first prediction, that the soil microbial community invests more energy in the production of
451
ecoenzymes to acquire nutrients in sites of low resource availability, such as the RS soil, was
452
confirmed. We observed that the RS soil showed a lower concentration of available P than the G
453
soil in the three years studied and, consequently, the RS soil microbial community invested
454
more energy in the acquisition of P (increased enzymatic activity of phosphomonoesterase and
455
phosphodiesterase) than the G soil microbial community only during the two wet years. In
456
contrast, the G soil had higher Cmic, Nmic and Pmic concentrations and lower enzymatic
457
activity of phosphomonoesterase and phosphodiesterase than the RS soil during both wet years,
458
which also supports our prediction (Fig 1 and Table 3). These results suggest that the microbial
459
community in the RS soil, with lower resource availability, must reduce growth as a result of: 1) the
460
physiological cost associated with a low reallocation to P-rich ribosomal RNA, as suggested by
461
the growth rate hypothesis (GRH) (Sinsabaugh & Follstad Shah 2012; Sterner & Elser 2002;
462
Zechmeister-Boltenstern et al. 2015) and 2) the required investment of energy towards the
463
acquisition of P in order to produce ecoenzymes (Evans & Wallenstein 2012; Schimel et al. 2007;
464
Wallenstein & Hall 2012). The microbial C:N:P ratio was greater in the RS soil (127:19:1) than in
465
the G soil (63:10:1), suggesting that the microbial community in the former site is more P-
466
constrained (Cleveland & Liptzin 2007). The studied soils are characterized by low P availability PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed Microbial community size begins to be limited by the availability of labile C, which produces
483
a change in the microbial community composition towards microbial guilds with lower growth
484
rates (low concentration of Cmic), but with the capacity to produce polyphenol oxidase to break
485
down structurally complex molecules and obtain C (Moorhead & Sinsabaugh 2006). This
486
situation is comparable to the conditions of the G soil in the wet year 2014, where the microbial
487
community was required to cleave lignin in order to maintain its growth rate. 488
Furthermore, the differences in soil nutrient dynamics between both sites can be strongly
489
affected by soil microbial composition. While analyses of soil bacterial composition were only PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 490
determined for 2011, this year presented the highest soil water availability and also showed
491
higher concentrations of Cmic, Nmic, and Pmic than in the other studied years. Several studies
492
(Nemergut et al. 2010; Philippot et al. 2009) have reported that heterotrophic decomposition
493
depends on the relative abundance of specific taxa because different species process organic
494
matter at different rates, even under similar soil conditions. The G soil had a higher proportion of
495
Proteobacterias, Actinobacterias and Bacteroidetes than the RS soil, and some species of these
496
taxa have the capacity to produce β-glucosidase (BG) (Moreno et al. 2013), cellobiohydrolase
497
(CBH), poliphenoloxidase (PPO) (T K Kirk & Farrell 1987), glucanases and glycosidases (Xie et
498
al. 2007), which act to cleave C molecules. 499
In contrast, the scrub soil had higher proportion of Acidobaterias and Firmicutes, including
500
species with the capacity for producing enzymes for P mineralization (Koch et al. 2008; Tan et
501
al. 2013). Moreover, the acidobacterias of the RS soil could contribute to the release of
502
unavailable P through organic acid release (Tan et al. 2013) and, together with Firmicutes, can
503
mineralize P via the production of phosphatases, as has been observed in isolates of
504
acidobacterias from substrates with low C concentrations (Koch et al. 2008; Tan et al. 2013). 505
Chloroflexi was present in a higher proportion in the RS than in the G soil, but both soils had a
506
similar proportion of Cyanobacteria suggesting that the amount of microbial desert crust is
507
similar in both sites. Both phyla are facultative autotrophic bacteria (Smith 1983) and therefore
508
have the capacity to fix atmospheric C and to produce coenzymes for depolymerization and
509
mineralization of C (Berg et al. 2010; Mitsui et al. 1986; Smith 1983). The Cyanobacteria also
510
have the capacity to fix atmospheric N. Fixation in the microbial biomass of C and N by these
511
taxa could represent an important input of both nutrients to the soil (Mitsui et al. 1986; Smith
512
1983). Wallenstein & Hall (2012) proposed that sites limited by nutrients are more vulnerable to 499
In contrast, the scrub soil had higher proportion of Acidobaterias and Firmicutes, including
500
species with the capacity for producing enzymes for P mineralization (Koch et al. 2008; Tan et
501
al. 2013). Manuscript to be reviewed 513
rainfall variability, because the microbial community must invest energy in nutrient acquisition,
514
thus reducing its capacity for adaptation required by fluctuation in water availability. We
515
proposed that sites with low resources availability, such as the RS soil, could be thus more
516
vulnerable to annual precipitation variability. 517
518
3.2. Resilience in the face of precipitation changes
519
Our second prediction, that the microbial community will be more vulnerable to variability in
520
precipitation in the site with lower soil resources (RS), was not confirmed because the soil
521
community was resilient to soil P coinstrains by ecoenzyme upregulation during times of
522
adequate moisture. In both vegetation cover types, nutrient availability increased with increased
523
precipitation. The correlation between precipitation and the Cmic, Nmic and Pmic, indicate that a
524
higher amount of rainfall favored the microbial immobilization of these nutrients under both
525
vegetation cover types. Nevertheless, compared to the RS soil, the G soil showed steeper slopes
526
in regressions between the precipitation and the concentrations of Nmic and ratios of Cmic:Pmic
527
and Nmic:Pmic (Table S1), suggesting that the microbial community of the grassland soil has the
528
ability to immobilize more N within its microbial biomass and more rapidly than the microbial
529
community of the RS soil. Positive correlations between Cmic and rainfall have been reported
530
for an oak forest (Baldrian et al. 2010) and a semiarid grassland (Zhou et al. 2013), but a
531
correlation between precipitation with Nmic and Pmic concentrations has not hitherto been
532
reported for natural ecosystems. 533
Furthermore, in the soil community homeostasis analyses, the relationships between log C:NR
d l
d b
l
d l
i
h
d h
il h d l
h
did 513
rainfall variability, because the microbial community must invest energy in nutrient acquisition,
514
thus reducing its capacity for adaptation required by fluctuation in water availability. We
515
proposed that sites with low resources availability, such as the RS soil, could be thus more
516
vulnerable to annual precipitation variability. 513
rainfall variability, because the microbial community must invest energy in nutrient acquisition,
514
thus reducing its capacity for adaptation required by fluctuation in water availability. We
515
proposed that sites with low resources availability, such as the RS soil, could be thus more
516
vulnerable to annual precipitation variability. Manuscript to be reviewed Moreover, the acidobacterias of the RS soil could contribute to the release of
502
unavailable P through organic acid release (Tan et al. 2013) and, together with Firmicutes, can
503
mineralize P via the production of phosphatases, as has been observed in isolates of
504
acidobacterias from substrates with low C concentrations (Koch et al. 2008; Tan et al. 2013). 505
Chloroflexi was present in a higher proportion in the RS than in the G soil, but both soils had a
506
similar proportion of Cyanobacteria suggesting that the amount of microbial desert crust is
507
similar in both sites. Both phyla are facultative autotrophic bacteria (Smith 1983) and therefore
508
have the capacity to fix atmospheric C and to produce coenzymes for depolymerization and
509
mineralization of C (Berg et al. 2010; Mitsui et al. 1986; Smith 1983). The Cyanobacteria also
510
have the capacity to fix atmospheric N. Fixation in the microbial biomass of C and N by these
511
taxa could represent an important input of both nutrients to the soil (Mitsui et al. 1986; Smith
512
1983). Wallenstein & Hall (2012) proposed that sites limited by nutrients are more vulnerable to PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 519
Our second prediction, that the microbial community will be more vulnerable to variability in
520
precipitation in the site with lower soil resources (RS), was not confirmed because the soil
521
community was resilient to soil P coinstrains by ecoenzyme upregulation during times of
522
adequate moisture. In both vegetation cover types, nutrient availability increased with increased
523
precipitation. The correlation between precipitation and the Cmic, Nmic and Pmic, indicate that a
524
higher amount of rainfall favored the microbial immobilization of these nutrients under both
525
vegetation cover types. Nevertheless, compared to the RS soil, the G soil showed steeper slopes
526
in regressions between the precipitation and the concentrations of Nmic and ratios of Cmic:Pmic
527
and Nmic:Pmic (Table S1), suggesting that the microbial community of the grassland soil has the
528
ability to immobilize more N within its microbial biomass and more rapidly than the microbial
529
community of the RS soil. Positive correlations between Cmic and rainfall have been reported
530
for an oak forest (Baldrian et al. 2010) and a semiarid grassland (Zhou et al. 2013), but a
531
correlation between precipitation with Nmic and Pmic concentrations has not hitherto been
532
reported for natural ecosystems. 533
Furthermore, in the soil community homeostasis analyses, the relationships between log C:NR
534
and log C:NB, and between log C:PR and log C:PB in the G and the RS soils had slopes that did
535
not differ significantly from zero (Figures S1 and S2), suggesting that the soil microbial PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 559
associated with P acquisition and elevated TERC:P values when the water is not limiting (2013
560
and 2014) suggest that the RS soil microbial community is well adapted to acquire P resources
561
via ecoenzyme upregulation post drought. 562
We suggested that, under the scenario proposed by Global Climate Change models for desert
563
ecosystems that predict reduced annual precipitation and increased rainfall variability, the
564
microbial community from both sites could be vulnerable to drought events, but the RS soil
565
microbial communities can make adjustments in order to obtain nutrients in wet years,
566
suggesting that this community is resilient post drought. 567
568
4. Conclusion
569
Soil communities of both sites (RS and G) may be vulnerable to drought. However, the
570
community at the site with lower resources (RS) may have evolved adaptations, such as rapid
571
ecoenzymatic upregulation, under chronic P limitation. This adaptation confers greater resilience
572
within the community to respond to precipitation events post drought. Under the Global Climate
573
Change scenarios for desert ecosystems that predict reduced annual precipitation and an
574
increased intensity and frequency of torrential rains and drought events, soil microbial
575
communities within both sites could be vulnerable to drought through the combination of C and
576
P co-limitation and reallocation of energy and nutrient resources to physiological acclimatization
577
strategies in order to survive. 578 559
associated with P acquisition and elevated TERC:P values when the water is not limiting (2013
560
and 2014) suggest that the RS soil microbial community is well adapted to acquire P resources
561
via ecoenzyme upregulation post drought. 559
associated with P acquisition and elevated TERC:P values when the water is not limiting (2013
560
and 2014) suggest that the RS soil microbial community is well adapted to acquire P resources
561
via ecoenzyme upregulation post drought. 562
We suggested that, under the scenario proposed by Global Climate Change models for desert
563
ecosystems that predict reduced annual precipitation and increased rainfall variability, the
564
microbial community from both sites could be vulnerable to drought events, but the RS soil
565
microbial communities can make adjustments in order to obtain nutrients in wet years,
566
suggesting that this community is resilient post drought. Manuscript to be reviewed 536
communities adjust physiologically (Sinsabaugh & Follstad Shah 2012) to processing low N and
537
P resources in order to cope with the nutrient limitation, particularly in dry years. Our data also
538
suggest that these physiological adjustments occurred differently in the soil microbial
539
communities of the two vegetation covers and was related to both precipitation quantity and
540
nutrient availability. 536
communities adjust physiologically (Sinsabaugh & Follstad Shah 2012) to processing low N and
537
P resources in order to cope with the nutrient limitation, particularly in dry years. Our data also
538
suggest that these physiological adjustments occurred differently in the soil microbial
539
communities of the two vegetation covers and was related to both precipitation quantity and
540
nutrient availability. 541
Our results show how values of TERC:N and TERC:P may shift with respect to variation in
542
annual rainfall and different vegetation cover. The estimated TERC:N was lower in the wet year
543
for both sites, indicating greater sensitivity to N limitation due to the rapid growth of the
544
microbial community produced by the water availability. For TERC:P, we observed site-specific
545
differences. The TERC:P was higher in the RS soil than in the G soil for 2013 and 2014, 546
indicating a greater sensitivity of the microbial community to P limitation in the G soil. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 562
We suggested that, under the scenario proposed by Global Climate Change models for desert
563
ecosystems that predict reduced annual precipitation and increased rainfall variability, the
564
microbial community from both sites could be vulnerable to drought events, but the RS soil
565
microbial communities can make adjustments in order to obtain nutrients in wet years,
566
suggesting that this community is resilient post drought. 568
4. Conclusion
569
Soil communities of both sites (RS and G) may be vulnerable to drought. However, the
570
community at the site with lower resources (RS) may have evolved adaptations, such as rapid
571
ecoenzymatic upregulation, under chronic P limitation. This adaptation confers greater resilience
572
within the community to respond to precipitation events post drought. Under the Global Climate
573
Change scenarios for desert ecosystems that predict reduced annual precipitation and an
574
increased intensity and frequency of torrential rains and drought events, soil microbial
575
communities within both sites could be vulnerable to drought through the combination of C and
576
P co-limitation and reallocation of energy and nutrient resources to physiological acclimatization
577
strategies in order to survive. 568
4. Conclusion
569
Soil communities of both sites (RS and G) may be vulnerable to drought. However, the
570
community at the site with lower resources (RS) may have evolved adaptations, such as rapid
571
ecoenzymatic upregulation, under chronic P limitation. This adaptation confers greater resilience
572
within the community to respond to precipitation events post drought. Under the Global Climate
573
Change scenarios for desert ecosystems that predict reduced annual precipitation and an
574
increased intensity and frequency of torrential rains and drought events, soil microbial
575
communities within both sites could be vulnerable to drought through the combination of C and
576
P co-limitation and reallocation of energy and nutrient resources to physiological acclimatization
577
strategies in order to survive. 580
This paper is presented by Cristina Montiel-González as partial fulfillment of a doctoral degree
581
at the “Programa de Posgrado en Ciencias Biológicas, UNAM”. We thank Velazquez-Rodrigo 580
This paper is presented by Cristina Montiel-González as partial fulfillment of a doctoral degree
581
at the “Programa de Posgrado en Ciencias Biológicas, UNAM”. We thank Velazquez-Rodrigo 586
REFERENCES 587
Allison SD, and Martiny JBH. 2008. Resistance, resilience, and redundancy in microbial
588
communities. Proceedings of the National Academy of Sciences 105:11512-11519. 589
Austin AT, Yahdjian L, Stark JM, Belnap J, Porporato A, Norton U, Ravetta DA, and Schaeffer
590
SM. 2004. Water pulses and biogeochemical cycles in arid and semiarid ecosystems. 591
Oecologia 141:221-235. 592
Baldrian P, Merhautová V, Petránková M, Cajthaml T, and Šnajdr J. 2010. Distribution of
593
microbial biomass and activity of extracellular enzymes in a hardwood forest soil reflect
594
soil moisture content. Applied Soil Ecology 46:177-182. 595
Bell CW, Acosta-Martinez V, McIntyre NE, Cox S, Tissue DT, and Zak JC. 2009. Linking
596
Microbial Community Structure and Function to Seasonal Differences in Soil Moisture
597
and Temperature in a Chihuahuan Desert Grassland. Microbial Ecology 58:827-842. 598
Bell CW, Tissue DT, Loik ME, Wallenstein MD, Acosta - Martinez V, Erickson RA, and Zak
599
JC. 2014. Soil microbial and nutrient responses to 7 years of seasonally altered
600
precipitation in a Chihuahuan Desert grassland. Global Change Biology 20:1657-1673. 601
Berg IA, Kockelkorn D, Ramos-Vera WH, Say RF, Zarzycki J, Hügler M, Alber BE, and Fuchs
602
G. 2010. Autotrophic carbon fixation in archaea. Nat Rev Micro 8:447-460. 603
Bremmer JM. 1996. Nitrogen-total. In: Sparks D, Page AL, Helmke PA, Loerppert RGH,
604
Soltanpour PN, Tabatabai MA, Jhonson CT, and Sumner ME, eds. Methods of soil
605
analysis part 3: chemical analysis. Madison, USA: Soil Sicence Society of American and
606
American Society of Agronomy, 1085-1121. 607
Brookes PC, Powlson DS, and Jenkinson DS. 1984. Phosphorus in the soil microbial biomass. 608
Soil Biology and Biochemistry 16:169-175. 609
Burns R, G. , De Forest J, L. , Marxsen J, Sinsabaugh R, L. , Stromberger M, E. , Wallenstein M,
610
D. , Weintraub M, N. , and Zoppini A. 2013. Soil enzymes in a changing environment:
611
Current knowledge and future directions. Soil Biology and Biochemistry 58. 612
Classen AT, Sundqvist MK, Henning JA, Newman GS, Moore JAM, Cregger MA, Moorhead
613
LC, and Patterson CM. 2015. Direct and indirect effects of climate change on soil
614
microbial and soil microbial-plant interactions: What lies ahead? Ecosphere 6:1-21. 615
Cleveland CC, and Liptzin D. 2007. C:N:P stoichiometry in soil: is there a “Redfield ratio” for
616
the microbial biomass? Biogeochemistry 85:235-252. 617
Coleman D, C. , and Whitman W, B. . 2005. Manuscript to be reviewed 582
for his assistance during chemical analyses and Angel Bravo-Monzón for his helpful comments
583
on earlier versions of this manuscript. We thank two anonymous reviewers for comments on a
584
draft of the manuscript 582
for his assistance during chemical analyses and Angel Bravo-Monzón for his helpful comments
583
on earlier versions of this manuscript. We thank two anonymous reviewers for comments on a
584
draft of the manuscript. 582
for his assistance during chemical analyses and Angel Bravo-Monzón for his helpful comments
583
on earlier versions of this manuscript. We thank two anonymous reviewers for comments on a Manuscript to be reviewed 622
MA. 2011. Temperature and soil organic matter decomposition rates – synthesis of
623
current knowledge and a way forward. Global Change Biology 17:3392-3404. 624
Cregger MA, Schadt CW, McDowell NG, Pockman WT, and Classen AT. 2012. Response of the
625
soil microbial community to changes in precipitation in a semiarid ecosystem. Applied
626
and environmental Microbiology 78:8587-8594. 627
Chavez-Vergara B, Merino A, Vázquez-Marrufo G, and García-Oliva F. 2014. Organic matter
628
dynamics and microbial activity during decomposition of forest floor under two native
629
neotropical oak species in a temperate deciduous forest in Mexico. Geoderma 235–
630
236:133-145. 631
D'Odorico P, and Bhattachan A. 2012. Hydrologic variability in dryland regions: impacts on
632
ecosystem dynamics and food security. Philosophical Transactions of the Royal Society
633
of London B: Biological Sciences 367:3145-3157. 634
D’Odorico P, Bhattachan A, Davis KF, Ravi S, and Runyan CW. 2013. Global desertification:
635
Drivers and feedbacks. Advances in Water Resources 51:326-344. 636
Eivazi F, and Tabatabai MA. 1977. Phosphatases in soils. Soil Biology and Biochemistry 9:167-
637
172. 638
Eivazi F, and Tabatabai MA. 1988. Glucosidases and galactosidases in soils. Soil Biology and
639
Biochemistry 20:601-606. 640
Ekschmitt K, Liu M, Vetter S, Fox O, and Wolters V. 2005. Strategies used by soil biota to
641
overcome soil organic matter stability — why is dead organic matter left over in the soil? 642
Geoderma 128:167-176. 643
Elser JJ, Schampel JH, Garcia-Pichel F, Wade BD, Souza V, Eguiarte L, Escalante ANA, and
644
Farmer JD. 2005. Effects of phosphorus enrichment and grazing snails on modern
645
stromatolitic microbial communities. Freshwater Biology 50:1808-1825. 646
Esch EH, Lipson D, and Cleland EE. 2016. Direct and indirect effects of shifting rainfall on soil
647
microbial respiration and enzyme activity in a semi-arid system. Plant and Soil:1-14. 648
Evans SE, and Wallenstein MD. 2012. Soil microbial community response to drying and
649
rewetting stress: does historical precipitation regime matter? Biogeochemistry 109:101-
650
116. 651
Farrell M, Prendergast-Miller M, Jones DL, Hill PW, and Condron LM. 2014. Soil microbial
652
organic nitrogen uptake is regulated by carbon availability. Soil Biology and
653
Biochemistry 77:261-267. 654
Fay PA, Kaufman DM, Nippert JB, Carlisle JD, and Harper CW. 2008. Changes in grassland
655
ecosystem function due to extreme rainfall events: implications for responses to climate
656
change. Global Change Biology 14:1600-1608. 586
REFERENCES Linking species richness, biodiversity and
618
ecosystem function in soil systems. Pedobiologia 49. 619
Conant RT, Ryan MG, Ågren GI, Birge HE, Davidson EA, Eliasson PE, Evans SE, Frey SD,
620
Giardina CP, Hopkins FM, Hyvönen R, Kirschbaum MUF, Lavallee JM, Leifeld J,
621
Parton WJ, Megan Steinweg J, Wallenstein MD, Martin Wetterstedt JÅ, and Bradford PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 668
Housman DC, Yeager CM, Darby BJ, Sanford Jr RL, Kuske CR, Neher DA, and Belnap J. 2007. 669
Heterogeneity of soil nutrients and subsurface biota in a dryland ecosystem. Soil Biology
670
and Biochemistry 39:2138-2149. 671
Huffman EWD. 1977. Performance of a new automatic carbon dioxide coulometer. 672
Microchemical Journal 22:567-573. 673
IPCC. 2013. Climate Change 2013: The Physical Science Basis. Contribution of Working Group
674
I to the Fifth Assessment Report of the Intergovernmental Panel on Climate Change
675
[Stocker,T.F., D. Qin, G.-K. Plattner, M. Tignor, S.K. Allen, J. Boschung, A. Nauels, Y. 676
Xia, V. Bex and P.M. Midgley (eds.)]. Cambridge University Press, Cambridge, United
677
Kingdom and New York, NY, USA. In: Stocker TF, D. Qin, G.-K. Plattner, M. Tignor,
678
S.K. Allen, J. Boschung, A. Nauels, Y. Xia, V. Bex and P.M. Midgley, editor. 679
Cambridge, United Kingdom and New York, NY, USA. 680
Joergensen RG. 1996. The fumigation-extraction method to estimate soil microbial biomass:
681
Calibration of the kEC value. Soil Biology and Biochemistry 28:25-31. 682
Joergensen RG, and Mueller T. 1996. The fumigation-extraction method to estimate soil
683
microbial biomass: Calibration of the kEN value. Soil Biology and Biochemistry 28:33-
684
37. 685
Johannes C, and Majcherczyk A. 2000. Laccase activity tests and laccase inhibitors. Journal of
686
Biotechnology 78:193-199. 687
Jones DL, and Willett VB. 2006. Experimental evaluation of methods to quantify dissolved
688
organic nitrogen (DON) and dissolved organic carbon (DOC) in soil. Soil Biology and
689
Biochemistry 38:991-999. 690
Jones DL, Willett VB, Stockdale EA, Macdonald AJ, and Murphy DV. 2012. Molecular Weight
691
of Dissolved Organic Carbon, Nitrogen, and Phenolics in Grassland Soils. Soil Science
692
Society of America Journal 76:142-150. 693
Karl DM. 2014. Microbially mediated transformations of phosphorus in the sea: new views of an
694
old cycle. Annual Review of Marine Science 6:279-337. 695
Koch IH, Gich F, Dunfield PF, and Overmann J. 2008. Edaphobacter modestus gen. nov., sp. 696
nov., and Edaphobacter aggregans sp. nov., acidobacteria isolated from alpine and forest
697
soils. International Journal of Systematic and Evolutionary Microbiology 58:1114-1122. 698
Lajtha K, Driscoll TC, Jarrell MW, and Edward TE. 1999. Soil phosphorus: characterization and
699
total element analysis. In: Robertson GP, Coleman CD, Bledsoe SC, and Sollins P, eds. 700
Soil Methods for Long-Term Ecological Research. Oxford University Press, New York
701
LTER, 115-142. Manuscript to be reviewed 657
Frost PC, Benstead JP, Cross WF, Hillebrand H, Larson JH, Xenopoulos MA, and Yoshida T. 658
2006. Threshold elemental ratios of carbon and phosphorus in aquatic consumers. 659
Ecology Letters 9:774-779. 660
González MF. 2012. Las zonas áridas y semiáridas de México y su vegetación. México, DF:
661
INE-SEMARNAT. 662
Henry HAL. 2013. Reprint of “Soil extracellular enzyme dynamics in a changing climate”. Soil
663
Biology and Biochemistry 56:53-59. 664
Holmgren M, Stapp P, Dickman CR, Gracia C, Graham S, Gutiérrez JR, Hice C, Jaksic F, Kelt
665
DA, Letnic M, Lima M, López BC, Meserve PL, Milstead WB, Polis GA, Previtali MA,
666
Richter M, Sabaté S, and Squeo FA. 2006. Extreme climatic events shape arid and
667
semiarid ecosystems. Frontiers in Ecology and the Environment 4:87-95. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 713
Moorhead DL, and Sinsabaugh RL. 2006. A THEORETICAL MODEL OF LITTER DECAY
714
AND MICROBIAL INTERACTION. Ecological Monographs 76:151-174. 715
Moreno B, Cañizares R, Nuñez R, and Benitez E. 2013. Genetic diversity of bacterial β-
716
glucosidase-encoding genes as a function of soil management. Biology and Fertility of
717
Soils 49:735-745. 718
Murphy J, and Riley JP. 1962. A modified single solution method for the determination of
719
phosphate in natural waters. Analytica Chimica Acta 27:31-36. 720
Nemergut DR, Cleveland CC, Wieder WR, Washenberger CL, and Townsend AR. 2010. Plot-
721
scale manipulations of organic matter inputs to soils correlate with shifts in microbial
722
community composition in a lowland tropical rain forest. Soil Biology and Biochemistry
723
42:2153-2160. 724
Perroni Y, García-Oliva F, and Souza V. 2014a. Plant species identity and soil P forms in an
725
oligotrophic grassland–desert scrub system. Journal of Arid Environments 108:29-37. 726
Perroni Y, García-Oliva F, Tapia-Torres Y, and Souza V. 2014b. Relationship between soil P
727
fractions and microbial biomass in an oligotrophic grassland-desert scrub system. 728
Ecological Research 29:463-472. 729
Philippot L, Bru D, Saby NPA, Čuhel J, Arrouays D, Šimek M, and Hallin S. 2009. Spatial
730
patterns of bacterial taxa in nature reflect ecological traits of deep branches of the 16S
731
rRNA bacterial tree. Environmental Microbiology 11:3096-3104. 732
Schimel J, Balser TC, and Wallenstein M. 2007. Microbial stress-response physiology and its
733
implications for ecosystem function. Ecology 88:1386-1394. p
y
gy
734
Schimel J, and Schaeffer SM. 2012. Microbial control over carbon cycling in soil. Frontiers in
735
Microbiology 3. 736
Singh B, Quince C, Macdonald C, Khachane A, Thomas N, Al-Soud W, Sørensen S, He Z,
737
White D, Sinclair A, Crooks B, Zhou J, and Campbell C. 2014. Loss of microbial
738
diversity in soils is coincident with reductions in some specialized functions. 739
Environmental Microbiology. 740
Sinsabaugh RL. 2010. Phenol oxidase, peroxidase and organic matter dynamics of soil. Soil
741
Biology and Biochemistry 42:391-404. gy
y
742
Sinsabaugh RL, and Follstad Shah JJ. 2011. Ecoenzymatic stoichiometry of recalcitrant organic
743
matter decomposition: the growth rate hypothesis in reverse. Biogeochemistry 102:31-43. 744
Sinsabaugh RL, and Follstad Shah JJ. 2012. Ecoenzymatic Stoichiometry and Ecological
745
Theory. Annual Review of Ecology, Evolution, and Systematics 43:313-343. 746
Sinsabaugh RL, Hill BH, and Follstad Shah JJ. 2009. Ecoenzymatic stoichiometry of microbial
747
organic nutrient acquisition in soil and sediment. Manuscript to be reviewed 702
López-Lozano NE, Eguiarte LE, Bonilla-Rosso G, García-Oliva F, Martínez-Piedragil C, Rooks
703
C, and Souza V. 2012. Bacterial Communities and the Nitrogen Cycle in the Gypsum
704
Soils of Cuatro Ciénegas Basin, Coahuila: A Mars Analogue. Astrobiology 12:699-709. 702
López-Lozano NE, Eguiarte LE, Bonilla-Rosso G, García-Oliva F, Martínez-Piedragil C, Rooks
703
C, and Souza V. 2012. Bacterial Communities and the Nitrogen Cycle in the Gypsum 705
Manzoni S, Schimel JP, and Porporato A. 2012. Responses of soil microbial communities to
706
water stress: results from a meta-analysis. Ecology 93:930-938. 707
McKee JW, Jones NW, and Long LE. 1990. Stratigraphy and provenance of strata along the San
708
Marcos fault, central Coahuila, Mexico. Geological Society of America Bulletin 102:593-
709
614. 710
Mitsui A, Kumazawa S, Takahashi A, Ikemoto H, Cao S, and Arai T. 1986. Strategy by which
711
nitrogen-fixing unicellular cyanobacteria grow photoautotrophically. Nature 323:720-
712
722. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 758
T K Kirk a, and Farrell RL. 1987. Enzymatic "Combustion": The Microbial Degradation of
759
Lignin. Annual Review of Microbiology 41:465-501. 760
Tabatabai MA, and Bremner JM. 1969. Use of p-nitrophenyl phosphate for assay of soil
761
phosphatase activity. Soil Biology and Biochemistry 1:301-307. 762
Tan H, Barret M, Mooij MJ, Rice O, Morrissey JP, Dobson A, Griffiths B, and O’Gara F. 2013. 763
Long-term phosphorus fertilisation increased the diversity of the total bacterial
764
community and the phoD phosphorus mineraliser group in pasture soils. Biology and
765
Fertility of Soils 49:661-672. 766
Tanenbaum DM, Goll J, Murphy S, Kumar P, Zafar N, Thiagarajan M, Madupu R, Davidsen T,
767
Kagan L, Kravitz S, Rusch DB, and Yooseph S. 2010. The JCVI standard operating
768
procedure for annotating prokaryotic metagenomic shotgun sequencing data. Standards
769
in Genomic Sciences 2:229-237. 770
Tapia-Torres Y, Elser JJ, Souza V, and García-Oliva F. 2015a. Ecoenzymatic stoichiometry at
771
the extremes: How microbes cope in an ultra-oligotrophic desert soil. Soil Biology and
772
Biochemistry 87:34-42. 773
Tapia-Torres Y, López-Lozano NE, Souza V, and García-Oliva F. 2015b. Vegetation-soil system
774
controls soil mechanisms for nitrogen transformations in an oligotrophic Mexican desert. 775
Journal of Arid Environments 114:62-69. 776
Tapia-Torres Y, Rodríguez-Torres MD, Elser JJ, Islas A, Souza V, García-Oliva F, and Olmedo-
777
Álvarez G. 2016. How to live with phosphorus scarcity in soil and sediment: Lessons
778
from bacteria. Applied and environmental Microbiology. 779
Technicon. 1977. Technicon
Industrial
System. Method
No. 329-74
W/B
780
Individual/simultaneous determinations of nitrogen and/or phosphorus in BD acid giest. 781
Analytical Chemistry 49:427A-427A. y
y
782
Thibault KM, and Brown JH. 2008. Impact of an extreme climatic event on community
783
assembly. Proceedings of the National Academy of Sciences 105:3410-3415. 784
Thomey ML, Collins SL, Vargas R, Johnson JE, Brown RF, Natvig DO, and Friggens MT. 2011. 785
Effect of precipitation variability on net primary production and soil respiration in a
786
Chihuahuan Desert grassland. Global Change Biology 17:1505-1515. g
g
gy
787
Tiessen H, and Moir JO. 2008. Characterization of available P by sequential extraction. In:
788
Carter MR, and Gregorich EG, eds. Soil sampling and methods of analysis. Second
789
Edition ed. Boca Raton Fl USA: CRC Press, 293-306. 790
Turner BL, Mahieu N, and Condron LM. 2003. Phosphorus-31 Nuclear Magnetic Resonance
791
Spectral Assignments of Phosphorus Compounds in Soil NaOH–EDTA Extracts. Manuscript to be reviewed Nature 462:795-798. 748
SMATR RDCT. 2007. R: a Language and Environment for Statistical Computing. R Foundation
749
for Statistical Computing, Vienna, ISBN 3-900051-07- 0. URL. 750
Smith AJ. 1983. Modes of cyanobacterial carbon metabolism. Annales de l'Institut Pasteur /
751
Microbiologie 134:93-113. 752
Souza V, Siefert JL, Escalante AE, Elser JJ, and Eguiarte LE. 2011. The Cuatro Ciénegas Basin
753
in Coahuila, Mexico: An Astrobiological Precambrian Park. Astrobiology 12:641-647. 754
Steinweg J, Dukes J, Eldor P, and Wallenstein M. 2013. Microbial responses to multi-factor
755
climate change: effects on soil enzymes. Frontiers in Microbiology 4:146. 756
Sterner RW, and Elser JJ. 2002. Ecological Stoichiometry: The Biology of Elements from
757
Molecules to the Biosphere: Princeton University Press. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 802
Wallenstein MD, and Hall EK. 2012. A trait-based framework for predicting when and where
803
microbial
adaptation
to
climate
change
will
affect
ecosystem
functioning. 804
Biogeochemistry 109:35-47. 802
Wallenstein MD, and Hall EK. 2012. A trait-based framework for predicting whe
803
microbial
adaptation
to
climate
change
will
affect
ecosystem
804
Biogeochemistry 109:35-47. 805
Waring BG, Weintraub SR, and Sinsabaugh RL. 2014. Ecoenzymatic stoichiometry
806
nutrient acquisition in tropical soils. Biogeochemistry 117:101-113. 807
Xie G, Bruce DC, Challacombe JF, Chertkov O, Detter JC, Gilna P, Han CS, Lucas
808
Myers GL, Richardson P, Tapia R, Thayer N, Thompson LS, Brettin TS,
809
Wilson DB, and McBride MJ. 2007. Genome Sequence of the Cellulol
810
Bacterium Cytophaga hutchinsonii. Applied and environmental Microbiolo
811
3546. 812
Zechmeister-Boltenstern S, Keiblinger KM, Mooshammer M, Peñuelas J, Richter A
813
and Wanek W. 2015. The application of ecological stoichiometry to plant–m
814
organic matter transformations. Ecological Monographs 85:133-155. 815
Zhou X, Chen C, Wang Y, Xu Z, Duan J, Hao Y, and Smaill S. 2013. Soil extrac
816
and nitrogen, microbial biomass and microbial metabolic activity in response
817
and increased precipitation in a semiarid Inner Mongolian grassland. Geode
818
31. 802
Wallenstein MD, and Hall EK. 2012. A trait-based framework for predicting when and where
803
microbial
adaptation
to
climate
change
will
affect
ecosystem
functioning. 804
Biogeochemistry 109:35-47. 805
Waring BG, Weintraub SR, and Sinsabaugh RL. 2014. Ecoenzymatic stoichiometry of microbial
806
nutrient acquisition in tropical soils. Biogeochemistry 117:101-113. 805
Waring BG, Weintraub SR, and Sinsabaugh RL. 2014. Ecoenzymatic stoichiometry of microbial
806
nutrient acquisition in tropical soils. Biogeochemistry 117:101-113. 807
Xie G, Bruce DC, Challacombe JF, Chertkov O, Detter JC, Gilna P, Han CS, Lucas S, Misra M,
808
Myers GL, Richardson P, Tapia R, Thayer N, Thompson LS, Brettin TS, Henrissat B,
809
Wilson DB, and McBride MJ. 2007. Genome Sequence of the Cellulolytic Gliding
810
Bacterium Cytophaga hutchinsonii. Applied and environmental Microbiology 73:3536-
811
3546. 812
Zechmeister-Boltenstern S, Keiblinger KM, Mooshammer M, Peñuelas J, Richter A, Sardans J,
813
and Wanek W. 2015. The application of ecological stoichiometry to plant–microbial–soil
814
organic matter transformations. Ecological Monographs 85:133-155. 815
Zhou X, Chen C, Wang Y, Xu Z, Duan J, Hao Y, and Smaill S. 2013. Soil extractable carbon
816
and nitrogen, microbial biomass and microbial metabolic activity in response to warming
817
and increased precipitation in a semiarid Inner Mongolian grassland. Manuscript to be reviewed Soil
792
Science Society of America Journal 67:497-510. 793
Vance ED, Brookes PC, and Jenkinson DS. 1987. An extraction method for measuring soil
794
microbial biomass C. Soil Biology and Biochemistry 19:703-707. 795
Verchot LV, and Borelli T. 2005. Application of para-nitrophenol (pNP) enzyme assays in
796
degraded tropical soils. Soil Biology and Biochemistry 37:625-633. 797
Von Ende NC. 2001. Repeated-measures Analysis. In: Scheiner MS, and Gurevitch J, eds. 798
Desing and Analysis of Ecological Experiments. Second Edition ed. New York: Oxford
799
University Press, 134-157. 800
Waldrop MP, Balser TC, and Firestone MK. 2000. Linking microbial community composition to
801
function in a tropical soil. Soil Biology and Biochemistry 32:1837-1846. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed Table 1(on next page) Manuscript to be reviewed Geoderma 206:24-
818
31. 819 PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Table 1 Means and (standard errors) of soil nutrients and ratios in the rosetophylous scrub (RS) and grassland (G)
soils over three consecutive years (2012, 2013 and 2014) in the Cuatro Ciénegas Basin, Coahuila, Mexico. Different uppercase letters (A and B) indicate significantly different means (P < 0.05) between vegetation
cover types (rosetophylous scrub and grassland) within the same sampling year (2012, 2013 and 2014);
whereas different lowercase letters (a, b and c) indicate significantly different means (P < 0.05) among
sampling dates within the same site. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed Table 2(on next page) Table 2(on next page) Manuscript to be reviewed 1
Table 1. Means and (standard errors) of soil nutrients and ratios in the rosetophylous scrub (RS)
2
and grassland (G) soils over three consecutive years (2012, 2013 and 2014) in the Cuatro
3
Ciénegas Basin, Coahuila, Mexico. Different uppercase letters (A and B) indicate significantly
4
different means (P < 0.05) between vegetation cover types (rosetophylous scrub and grassland)
5
within the same sampling year (2012, 2013 and 2014); whereas different lowercase letters (a, b
6
and c) indicate significantly different means (P < 0.05) among sampling dates within the same
7
site. 8 8 Year
2012
2013
2014
RS
G
RS
G
RS
G
Moisture (%)
12.7
(1.1)Bc
24.6
(2.5)Ab
24.6(3)Bab
43.5
(1.3)Aa
16.4(1.0)B
b
37.1(7.1)
Aa
pH
8.5 (0.06)
Aa
8.3
(0.04)Ba
8.1
(0.03)Ab
8.1
(0.02)Ab
8.1
(0.02)Ab
8.1
(0.1)Aab
Dissolved organic nutrient concentration
DOC (µg g-1)
9 (1)Ab
19 (4)Ac
23 (4) Ba
52 (1)Ab
28 (2)Ba
67 (4)Aa
DON (µg g-1)
4.1 (0.5)Bb
7.0 (0.6)Ab
5.5
(0.5)Bab
10.8
(0.8)Aa
6.9 (0.3)Ba
7.8
(0.4)Aab
DOP (µg g-1)
1.2 (0.2)Ab
0.3 (0.3)Ab
2.8 (0.2)Ba
5.1 (0.2)Aa
2.8 (0.2)Ba
5.3 (0.5)Aa
DOC:DON
2.3 (0.6)
3.1 (0.6)
4.2 (0.6)
4.9 (0.6)
4.0 (0.6)
6.8 (0.6)
DOC:DOP
7.9 (3.6)
15.3 (3.6)
8.2 (1.3)
18.8 (1.3)
10.3 (1.2)
13.0 (1.1)
DON:DOP
3.6 (0.4)
6.8 (3.3)
2.0 (0.1)
2.1 (0.1)
2.6 (0.4)
1.5 (0.1)
Available nutrient concentration
NH4 + (µg g-1)
2.8 (0.2)Ba
6.3 (0.5)Ac
3.6 (0.2)Ba
11.8
(1.1)Aa
2.7 (0.4)Ba
8. 9
(0.2)Ab
NO3- (µg g-1)
10.4
(1.4)Aa
6.7 (1.4)Ba
1.7 (0.3)Ab
3.2
(0.4)Aab
1.7 (0.1)Ab
1.0 (0.1)Ab
Pi (µg g-1)
1.9 (0.2)
2.5 (0.2)
2.9 (0.4)
4.5 (0.4)
3.9 (0.6)
5.3 (0.6)
9
DOC: dissolved organic carbon; DON: dissolved organic nitrogen; DOP: dissolved organic
10
phosphorus; NH4+: available ammonium; NO3-: available nitrate; Pi: Available inorganic
11
phosphorus. 13 13 PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Table 2 F-ratios and significant levels of the repeated-measures ANOVA for soil variables quantified in
the rosetophylous scrub and grassland soils over three consecutive years (2012, 2013 and
2014) in Cuatro Ciénegas Basin, Coahuila Mexico. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed Manuscript to be reviewed 1
Table 2. F-ratios and significant levels of the repeated-measures ANOVA for soil variables
2
quantified in the rosetophylous scrub and grassland soils over three consecutive years (2012,
3
2013 and 2014) in Cuatro Ciénegas Basin, Coahuila Mexico. 1
Table 2. F-ratios and significant levels of the repeated-measures ANOVA for soil variables
2
quantified in the rosetophylous scrub and grassland soils over three consecutive years (2012,
3
2013 and 2014) in Cuatro Ciénegas Basin, Coahuila Mexico. Source of variation
Between subjects
Within subjects
Parameters
Vegetation cover
Year
Vegetation cover X Year
Moisture
90.7 (<0.0001)
49.1(<0.0001)
2.7 (0.08)
pH
7.3 (0.02)
28.0 (<0.0001)
5.4 (0.01)
Dissolved nutrients
DOC
102.1(<0.0001)
79.2(<0.0001)
14.5 (<0.0001)
DON
38.5 (<0.0001)
25.1 (<0.0001)
3.8 (0.03)
DOP
14.1 (0.002)
55.0(<0.0001)
13.2 (0.0001)
DOC:DON
6.4 (0.02)
11.6 (0.0002)
2.0 (0.1)
DOC:DOP
9.1 (0.01)
0.5 (0.6)
1.8 (0.2)
DON:DOP
1.8 (0.2)
3.0 (0.07)
1.2 (0.3)
Available nutrients
NH4 +
236.8(<0.0001)
19.0 (<0.000)
10.5 (0.0005)
NO3-
1.8 (0.1)
47 (<0.0001)
5.4 (0.01)
Pi
14.2 (0.003)
12.9 (0.002)
1.1 (0.3)
4
DOC: dissolved organic carbon; DON: dissolved organic nitrogen; DOP: dissolved organic
5
phosphorus; NH4+: available ammonium; NO3-: available nitrate; Pi: Available inorganic
6
phosphorus. 8 PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed Table 3(on next page) Table 3(on next page) Table 3(on next page) 9
Cmic: microbial carbon; Nmic: microbial nitrogen; Pmic: microbial phosphorus. Manuscript to be reviewed 1
Table 3. Means and (standard errors) of microbial biomass nutrients, and microbial nutrient
2
ratios in the rosetophylous scrub (RS) and the grassland (G) soils over three consecutive years
3
(2012, 2013 and 2014) in the Cuatro Ciénegas Basin, Coahuila, Mexico. Different uppercase
4
letter (A and B) indicate that means differ significantly (P < 0.05) between vegetation cover
5
types (RS and G) within the same sampling year (2012, 2013 and 2014); whereas different
6
lowercase letters (a, b and c) indicate significantly different means (P < 0.05) among sampling
7
dates within the same site. Year
2012
2013
2014
RS
G
RS
G
RS
G
Nutrients concentration within microbial biomass
Cmic (μg g-1)
68 (12)
93 (12)
191 (20)
289 (20)
287 (16)
379 (1)
Nmic (μg g-1)
4.2
(0.6)Ab
6.4 (0.6)Ac
10.0
(1.0)Bb
22.0
(2.8)Ab
42.2
(1.7)Ba
59.8
(1.9)Aa
Pmic (μg g-1)
2.3
(0.6)Aa
2.5 (1.3)Ab
2.4 (0.1)Ba
6.4 (0.6)Aa
2.2
(0.02)Ba
6.1 (0.4)Aa
Cmic:Nmic
20 (4)
15 (4)
20 (2)
14 (2)
7 (0.3)
6 (0.3)
Cmic:Pmic
17 (5)Ac
9 (5)Ac
79 (6)Ab
48 (6)Bb
127
(0.2)Aa
63(4)Ba
Nmic:Pmic
0.9
(0.3)Ac
0.4 (0.2)Ac
4.2 (0.4)Ab
3.6 (0.6)Ab
18.7
(0.8)Aa
10.1
(0.7)Ba
8 10 Table 3 Means and (standard errors) of microbial biomass nutrients, and microbial nutrient ratios in
the rosetophylous scrub (RS) and the grassland (G) soils over three consecutive years (2012,
2013 and 2014) in the Cuatro Ciénegas Basin, Coahuila, Mexico. Different uppercase letter (A
and B) indicate that means differ significantly (P < 0.05) between vegetation cover types (RS
and G) within the same sampling year (2012, 2013 and 2014); whereas different lowercase
letters (a, b and c) indicate significantly different means (P < 0.05) among sampling dates
within the same site. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed Manuscript to be reviewed Table 4(on next page) Table 4(on next page) PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) 6
Cmic: microbial carbon; Nmic: microbial nitrogen; Pmic: microbial phosphorus; BG: β-1,4-
7
glucosidase; CBH: cellobiohydrolase; NAG: β-1,4-N-acetylglucosaminidase; PPO: polyphenol
8
oxidase; PME: phosphomonoesterase; PDE: phosphodiesterase Table 4 F-ratios and significant levels of the repeated measures ANOVA for microbial nutrient
concentration, microbial nutrient ratios and specific enzymatic activity quantified in the
rosetophylous scrub (RS) and the grassland (G) soils over three consecutive years (2012,
2013 and 2014) in Cuatro Ciénegas Basin, Coahuila Mexico. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed Manuscript to be reviewed 1
Table 4. F-ratios and significant levels of the repeated measures ANOVA for microbial nutrient
2
concentration, microbial nutrient ratios and specific enzymatic activity quantified in the
3
rosetophylous scrub (RS) and the grassland (G) soils over three consecutive years (2012, 2013
4
and 2014) in Cuatro Ciénegas Basin, Coahuila Mexico. 1
Table 4. F-ratios and significant levels of the repeated measures ANOVA for microbial nutrient
2
concentration, microbial nutrient ratios and specific enzymatic activity quantified in the
3
rosetophylous scrub (RS) and the grassland (G) soils over three consecutive years (2012, 2013
4
and 2014) in Cuatro Ciénegas Basin, Coahuila Mexico. p y
(
)
g
( )
y
(
,
4
and 2014) in Cuatro Ciénegas Basin, Coahuila Mexico. Source of variation
Between subject
Within subjects
Parameters
Vegetation cover
Year
Vegetation cover X Year
Dissolved nutrients
Cmic
62.1 (<0.0001)
93.3(<0.0001)
2.3 (0.11)
Nmic
48.7 (<0.0001)
484 (<0.0001)
12.9 (0.0001)
Pmic
24.6 (0.0003)
5.8 (0.008)
5.7 (0.009)
Cmic:Nmic
4.0 (0.07)
12.3 (0.0002)
0.7 (0.5)
Cmic:Pmic
107 (<0.0001)
92 (<0.0001)
11 (0.0005)
Nmic:Pmic
42 (<0.0001)
316 (<0.0001)
34 (<0.0001)
Specific enzymatic activity
BG
1.2 (0.28)
22.8 (<0.0001)
1.1 (0.33)
CBH
3 (0.1)
9.9 (<0.0001)
0.2 (0.7)
NAG
8.1(0.01)
52 (<0.0001)
10.8 (<0.0001)
PPO
8.8 (0.011)
34 (<0.0001)
4 (0.03)
PME
137 (<0.0001)
444(<0.0001)
80 (<0.0001)
PDE
67 (<0.0001)
232 (<0.0001)
19 (<0.0001)
5 Manuscript to be reviewed Figure 1(on next page) PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) The dotted line represents the standard deviation at 0.95. Figure 1(on next page) Figure 1(on next page) Simple linear regressions between annual accumulated precipitation before the
sampling date for four years and nutrients immobilized by microbial biomass for RS soil
and G soil. The dotted line represents the standard deviation at 0.95. Manuscript to be reviewed Figure 2(on next page) Figure 1(on next page) PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
0
200
400
600
800
1000
1200
G Cmic µg /g
y = 3.4031x -301.5226
R2=0.831532
p< 0.05
B)
Manuscript to be reviewed 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
0
10
20
30
40
50
60
70
G Nmic µg /g
y = 0.1476x + 0.5771
R² = 0.429
p= 0.0001
D)
5
6
7
8
9
10
mic µg /g
y = 0.0077x + 1.1766
R² = 0.2933
p=0.0029
E)
5
6
7
8
9
10
ic µg /g
y = 0.0125x + 2.4839
R² = 0.2339
p= 0.0091
F)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
-200
0
200
400
600
800
1000
1200
RS Cmic µg /g
y = 2.9606x - 293.77
R² = 0.7664
p<0.005
A)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
0
200
400
600
800
1000
1200
G Cmic µg /g
y = 3.4031x -301.5226
R2=0.831532
p< 0.05
B)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
0
10
20
30
40
50
60
70
RS Nmic µg /g
y = 0.074x + 3.3776
R² = 0.1995
p=0.0172
C)
Manuscript to be reviewed B) 0
240
260
280
300
320
340
360
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
0
200
400
600
800
1000
1200
G Cmic µg /g
y = 3.4031x -301.5226
R2=0.831532
p< 0.05
B)
Manuscript to be reviewed 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
-200
0
200
400
600
800
1000
1200
RS Cmic µg /g
y = 2.9606x - 293.77
R² = 0.7664
p<0.005
A)
60
80
100
120
140
160
180
200
220
240
260
2
0
200
400
600
800
1000
1200
G Cmic µg /g
y = 3.4031x -301.5226
R2=0.831532
p< 0.05
B)
Manuscript to be reviewed A) 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
-200
0
200
400
600
800
1000
1200
RS Cmic µg /g
y = 2.9606x - 293.77
R² = 0.7664
p<0.005
A)
G Cmic µg /g
B RS Cmic µg /g G Cmic µg /g RS 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
0
10
20
30
40
50
60
70
G Nmic µg /g
y = 0.1476x + 0.5771
R² = 0.429
p= 0.0001
D)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
Precipitation (mm)
-1
0
1
2
3
4
5
6
7
8
9
10
RS Pmic µg /g
y = 0.0077x + 1.1766
R² = 0.2933
p=0.0029
E)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
Precipitation (mm)
-1
0
1
2
3
4
5
6
7
8
9
10
G Pmic µg /g
y = 0.0125x + 2.4839
R² = 0.2339
p= 0.0091
F)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
0
10
20
30
40
50
60
70
RS Nmic µg /g
y = 0.074x + 3.3776
R² = 0.1995
p=0.0172
C)
PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) D
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
0
10
20
30
40
50
60
70
RS Nmic µg /g
y = 0.074x + 3.3776
R² = 0.1995
p=0.0172
C) 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
0
10
20
30
40
50
60
70
G Nmic µg /g
y = 0.1476x + 0.5771
R² = 0.429
p= 0.0001
D)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
0
10
20
30
40
50
60
70
RS Nmic µg /g
y = 0.074x + 3.3776
R² = 0.1995
p=0.0172
) 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
0
10
20
30
40
50
60
70
G Nmic µg /g
y = 0.1476x + 0.5771
R² = 0.429
p= 0.0001
D) C) D) y = 0.074x + 3.3776
R² = 0.1995
p=0.0172 RS Nmic µg /g G Nmic µg /g 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
Precipitation (mm)
-1
0
1
2
3
4
5
6
7
8
9
10
RS Pmic µg /g
y = 0.0077x + 1.1766
R² = 0.2933
p=0.0029
E)
PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
Precipitation (mm)
-1
0
1
2
3
4
5
6
7
8
9
10
RS Pmic µg /g
y = 0.0077x + 1.1766
R² = 0.2933
p=0.0029
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
Precipitation (mm)
-1
0
1
2
3
4
5
6
7
8
9
10
G Pmic µg /g
y = 0.0125x + 2.4839
R² = 0.2339
p= 0.0091
F)
PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
Precipitation (mm)
-1
0
1
2
3
4
5
6
7
8
9
10
G Pmic µg /g
y = 0.0125x + 2.4839
R² = 0.2339
p= 0.0091
F) E) F) RS Pmic µg /g 0
120
140
160
180
200
220
240
260
280
300
320
340
360
PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Precipitation (mm)
rJ reviewing PDF | (2017:05:17 Figure 2(on next page) Figure 2(on next page) Simple linear regressions between the annual accumulated precipitation before the
sampling date for four years and ratios of nutrients immobilized by microbial biomass
for RS soil and G soil. Figure 3(on next page) Figure 3(on next page) Manuscript to be reviewed Figure 2(on next page) PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
4
6
8
10
12
14
16
18
20
22
24
26
28
30
32
34
G Cmic/Nmic
y = 0.0295x + 8.4326
R² = 0.1305
p= 0.058
B)
Manuscript to be reviewed 8
10
12
14
Pmic
y = 0.028x - 0.7109
R² = 0.4646
p< 0.005
F)
12
14
16
18
20
22
24
/Pmic
y = 0.0219x +2.5719
R2= 0.083543
p= 0.135
E)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
-20
0
20
40
60
80
100
120
RS Cmic/Nmic
y = 0.1009x + 1.6994
R² = 0.2216
P=0.0115
A)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
4
6
8
10
12
14
16
18
20
22
24
26
28
30
32
34
G Cmic/Nmic
y = 0.0295x + 8.4326
R² = 0.1305
p= 0.058
B)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
-50
0
50
100
150
200
250
300
350
RS Cmic/Pmic
y = 0.7078x - 47.985
R² = 0.8131
p<0.005
C)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
-20
0
20
40
60
80
100
120
140
160
180
200
220
240
260
G Cmic/Pmic
y = 0.6341x - 63.665
R² = 0.8144
p<0.005
D)
Manuscript to be reviewed A) 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
-20
0
20
40
60
80
100
120
RS Cmic/Nmic
y = 0.1009x + 1.6994
R² = 0.2216
P=0.0115
A)
B RS Cmic/Nmic G Cmic/Nmic 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
Precipitation (mm)
-2
0
2
4
6
8
10
12
14
G Nmic/Pmic
y = 0.028x - 0.7109
R² = 0.4646
p< 0.005
F)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
Precipitation (mm)
-2
0
2
4
6
8
10
12
14
16
18
20
22
24
RS Nmic/Pmic
y = 0.0219x +2.5719
R2= 0.083543
p= 0.135
E)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
0
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
-50
0
50
100
150
200
250
300
350
RS Cmic/Pmic
y = 0.7078x - 47.985
R² = 0.8131
p<0.005
C)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
-20
0
20
40
60
80
100
120
140
160
180
200
220
240
260
G Cmic/Pmic
y = 0.6341x - 63.665
R² = 0.8144
p<0.005
D)
PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
y = 0.7078x - 47.985
R² = 0.8131
p<0.005
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
-20
0
20
40
60
80
100
120
140
160
180
200
220
240
260
G Cmic/Pmic
y = 0.6341x - 63.665
R² = 0.8144
p<0.005
D) 0
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
-20
0
20
40
60
80
100
120
140
160
180
200
220
240
260
G Cmic/Pmic
y = 0.6341x - 63.665
R² = 0.8144
p<0.005
D) 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
-50
0
50
100
150
200
250
300
350
RS Cmic/Pmic
y = 0.7078x - 47.985
R² = 0.8131
p<0.005
C)
G Cmic/Pmic
D D) C) y = 0.7078x - 47.985
R² = 0.8131
p<0.005 RS Cmic/Pmic G Cmic/Pmic 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
Precipitation (mm)
-2
0
2
4
6
8
10
12
14
G Nmic/Pmic
y = 0.028x - 0.7109
R² = 0.4646
p< 0.005
F)
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
Precipitation (mm)
-2
0
2
4
6
8
10
12
14
16
18
20
22
24
RS Nmic/Pmic
y = 0.0219x +2.5719
R2= 0.083543
p= 0.135
)
PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) G Nmic/Pmic
F
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
Precipitation (mm)
-2
0
2
4
6
8
10
12
14
16
18
20
22
24
RS Nmic/Pmic
y = 0.0219x +2.5719
R2= 0.083543
p= 0.135
E)
PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
Precipitation (mm)
-2
0
2
4
6
8
10
12
14
G Nmic/Pmic
y = 0.028x - 0.7109
R² = 0.4646
p< 0.005
F) 60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
Precipitation (mm)
-2
0
2
4
6
8
10
12
14
G Nmic/Pmic
y = 0.028x - 0.7109
R² = 0.4646
p< 0.005
F) E) F) RS Nmic/Pmic G Nmic/Pmic Precipitation (mm)
rJ reviewing PDF | (2017:05:17 Manuscript to be reviewed Specific enzymatic activity Specific enzymatic activity of: (A) β-1,4-glucosidase (BG), (B) cellobiohydrolase (CBH), (C) polyphenol
oxidase (PPO), (D) β-1,4-N-acetylglucosaminidase (NAG), (E) phosphomonoesterase (PME) and (F)
phosphodiesterase (PDE) in the rosetophylous scrub (RS) and grassland (G) soils over three consecutive
years (2012, 2013 and 2014) in the Cuatro Ciénegas Basin, Coahuila, Mexico. Different uppercase letters (A
and B) indicate significantly different means (P < 0.05) between vegetation cover types (RS and G) within
the same sampling year (2012, 2013 and 2014); whereas different lowercase letters (a, b and c) vertically
indicate significantly different means (P < 0.05) among sampling dates within the same site. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed Manuscript to be reviewed Manuscript to be reviewed Manuscript to be reviewed 2012
2013
2014
0.1
0.2
0.3
0.4
0.5
0.6
SEA BG (µmol mgCmic-1 h-1)
b
c
a
(A)
2012
2013
2014
0.04
0.06
0.08
0.10
0.12
0.14
0.16
0.18
SEA CBH (µmol mgCmic-1 h-1)
(B)
b
a
a
2012
2013
2014
-10
-5
0
5
10
15
20
25
30
35
40
45
SEA PPO (µmol mgCmic-1 h-1)
Aa
Ba
Aa
(C)
Ab
Aa
Aa
2012
2013
2014
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
SEA NAG (µmol mgNmic-1 h-1)
Aa
Ab
Ac
Ab
Aa
(D)
Ba
2012
2013
2014
YEAR
-5
0
5
10
15
20
25
30
35
40
45
SEA PME (µmol mgPmic-1 h-1)
(E)
Aa
Ab
Ab
Ba
Aa
Ba
2012
2013
2014
YEAR
-50
0
50
100
150
200
250
SEA PDE (µmol mgPmic-1 h-1)
Aa
(F)
Ab Ab
Aa
Ba
Ba
RS
G
SA
p 2012
2013
2014
0.1
0.2
0.3
0.4
0.5
0.6
SEA BG (µmol mgCmic-1 h-1)
b
c
a
(A) 2012
2013
2014
0.04
0.06
0.08
0.10
0.12
0.14
0.16
0.18
SEA CBH (µmol mgCmic-1 h-1)
(B)
b
a
a 2012
2013
2014
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
SEA NAG (µmol mgNmic-1 h-1)
Aa
Ab
Ac
Ab
Aa
(D)
Ba 2012
2013
2014
-10
-5
0
5
10
15
20
25
30
35
40
45
SEA PPO (µmol mgCmic-1 h-1)
Aa
Ba
Aa
(C)
Ab
Aa
Aa SA PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed Figure 5(on next page) Figure 4(on next page) Figure 4(on next page) Taxonomic distribution of sequences obtained from Pyrosequencing of 16S rRNA tags of
rosetophylous scrub and grassland soils during a wet year (2010). PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Manuscript to be reviewed 0
10
20
30
40
50
60
70
80
90
100
Rosetophylous scrub
Grassland
e Sequences
Unclassified
WS3
Verrucomicrobia
TM7
Synergistetes
Spirochaetes
Proteobacteria
Planctomycetes
OP11
OP10
OD1
Nitrospira
Gemmatimonadetes
Fusobacteria
Firmicutes
Deinococcus-Thermus
Deferribacteres
Cyanobacteria
Chloroflexi
Chlorob
Chlamydiae
Bacteroidetes
BRC1
Actinobacterias
Acidobacteria 100 60 Rosetophylous scrub Rosetophylous scrub Grassland PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) Figure 5(on next page) Figure 5(on next page) Threshold Elemental Ratio C:N and C:P (A and B, respectively) of the soil microbial
community over three consecutive years (2012, 2013 and 2014. PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017) 0
5
10
15
20
25
30
35
2012
2013
2014
TER C:N
Rosetophylous scrub
Grassland
Aa
Aa
Aa
Aa
Ab
Ab
A)
Manuscript to be reviewed A) Rosetophylous scrub
Grassland 0
20
40
60
80
100
120
140
2012
2013
2014
TER C:P
Ab
Aab
Aa
Bb
Aa
Ba
B)
0
5
10
15
20
25
30
35
2012
2013
2014
TER C:N
Aa
Aa
Aa
Aa
Ab
Ab
PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017)
Manuscript to be reviewed B) 0
20
40
60
80
100
120
2012
2013
2014
TER C:P
Ab
Aab
Aa
Bb
Ba
PeerJ reviewing PDF | (2017:05:17808:2:0:NEW 21 Sep 2017)
|
https://openalex.org/W4391283181
|
http://anapub.co.ke/journals/jcims/jcims_pdf/2024/jcims_volume_02/JCIMS202402001.pdf
|
English
| null |
Energy Utilization and Conversion in Modern Biomass Conversion Technologies
|
Journal of computational intelligence in materials science
| 2,024
|
cc-by
| 9,598
|
ISSN: 2959-832X ISSN: 2959-832X Journal of Computational Intelligence in Materials Science 2(2024) Correspondence should be addressed to Nancy Jan Sliper : nancyjan@aol.com. Correspondence should be addressed to Nancy Jan Sliper : nancyjan@aol.com. Article Info Journal of Computational Intelligence in Materials Science (https://anapub.co.ke/journals/jcims/jcims.html)
Doi: https://doi.org/10.53759/832X/JCIMS202402001. p
g
Received 10 September 2023; Revised from 02 November 2023; Accepted 12 December 2023. Available online 06 January 2024. ©2024 The Authors. Published by AnaPub Publications. hi i
i l
d
h CC
C
li This is an open access article under the CC BY-NC-ND license. (http://creativecommons.org/licenses/by Abstract – This paper provides a review on the current state of biomass conversion technologies that are in use and those
that could play a significant role in the future, such as those that might be linked to carbon dioxide (CO2) collection and
sequestered technology. Since the transportation industry is poised to become the most important new market for large-
scale efficient biomass usage, here is where most of the focus will be placed. Bio-energy contribution, now estimated at
40EJ to 55 EJ per year, is expected to expand significantly in the future. Nevertheless, the precise objective of bio-energy
will be dependent on the competitiveness aspect with bio-fuels and on agriculture policy globally. For the rest of this
century as least, observations suggest a range of 200–300 EJ, rendering biomass a more significant alternatives of energy
supply compared to mineral oil. The need to update bio-energy practices so they are compatible with sustainable
development strategies is a major concern. It is expected that within the next two to three decades, the cost of electricity
generated using sophisticated conversion concepts (such as gasification and contemporary co-firing and gasification) and
contemporary biomass sourced fuels (e.g., hydrogen, methanol, and ethyl alcohol from the lignocellulosic biomass) will
be competitive with conventional energy sources (partly based on price development with petroleum). An even more
efficient and cost-effective biofuel production system may be developed from sugarcane-centric ethanol within the
tropical climates. Keywords – Biomass conversion, co-firing, combustion, bio-oil extraction, gasification, digestion. Energy Utilization and Conversion in Modern
Biomass Conversion Technologies Nancy Jan Sliper
Advanced Chemical Metallurgy, Norwegian University of Science and Technology, Trondheim, Norway
nancyjan@aol.com I.
INTRODUCTION The development of large-scale biomass raw materials supply frameworks may be facilitated by flexible energy systems
that allow for the use of both biomass and fossil fuels. This might result in a reduced-cost, reduced-carbon, and reduced-
risk emission of energy in supply systems. The gasification pathway provides unique opportunities to couple this with low-
cost CO2 storage (and collection), leading to ideas that are versatile in terms of primary fuel input and product mix and
have the potential to achieve zero or negative carbon emissions. This can only be accomplished by persistent research and
development (R&D) initiatives, biomass market growth, sustained legislative support, and international cooperation. At the
present day, bio-fuels account for the significant portion of the globe’s energy distribution (about 80% of the total usage,
or more than 400 EJ annually). Biomass resources, however, meet 10-15% (or 45±10 EJ) of this need, making biomass the
most major renewable energy source to date. Biomass accounts for around 9-13% of overall energy sources in developed
nations, but as much as 25%-33% in poor countries [1]. In certain nations, biomass provides as much as 50-90% of the country's overall energy requirement. Yet, much of this
biomass use is domestic, with the poor relying on it for cooking and warmth. As a result, it is difficult to quantify
biomass's role in the energy supply, since its non-industrial application is poorly documented. Certain traditional uses are
also unsustainable since they deplete local soils of nutrients, lead to air and water pollution, and negatively affect people's
health. In case biomass is collected for pwer without compensatory reforestation and other practices for conservations, it
might potentially increase greenhouse gas emissions and have an impact on ecosystems. Even though some of this use is
commercial—for example, residential firewood in industrialized nations and charcoal (bio-char) and wood in urbanized
and commercial regions in developing nations—almost little information is available on the extent of these markets. There
are around 9.6 expected joules worth of stuff in these markets. Commercial power generation from biomass for application in energy production, industry, or transport fuel accounts
for a smaller but rising portion of modern bio-energy (some 7 EJ/yr in 2000). II.
TECHNOLOGY BACKGROUND OF BIOMASS CONVERSION Technological evaluation into the establishment of alternative sources of energy from bio-materials has been stimulated by
the environmental impacts of continuous fossil fuel consumption and Greenhouse Gases (GHGs). Carbon dioxide (CO2) is
the primary contributor to the increasing levels of greenhouse gas emissions in the atmosphere. In addition, the globe's
energy consumption is rising quickly, with bio-fuels contributing for more than 87% of all power production right now. Most organic energy resources (such as gas and oil reserves) are located in politically unstable locations, making
energy security a major concern. Thus, biogas from trash and leftovers may be an important part of the future energy
picture. Renewable and versatile, biogas may be used in lieu of fossil fuels in the production of power and heat and as a
gas fuel in vehicles. Moreover, biomethane (improved biogas) may replace natural gas in chemical manufacturing. Anaerobic digestion (AD) is an energy-efficient and ecologically beneficial technique, and recent assessments have shown
that the biogas it produces has substantial benefits over other sources of bioenergy [4]. By making use of readily accessible resources, AD technology may lower GHG emissions in contrast to fossil fuels. The technology's waste product, digestate, may be used as a high-value alternative to mineral fertilizers in crop production. As of 2014, the total amount of biogas produced in Europe was 1.35 × 107 t. As a result of the significant development of
agricultural biogas farm plants, Germany is the frontrunner in worldwide biogas generation, with approximately 25% of
the total integrated capacity. More than 8,000 installations for producing agricultural biogas were active in Germany by the
end of 2014 [5]. Numerous nations are now investing in research into alternative methods of creating biogas from
agricultural byproducts and municipal solid waste. The infrastructure in several European nations is now conducive to the
generation of biogas-powered energy. Remarkably, in Europe there is as much as 1.5 × 109 t of agro-biomass accessible
for AD. Biogas from cellulosic resources is a promising new energy source, and the United States, China, and India are all
investing in alternative production systems. While waste-derived biogas (and/or biomethane) shows promise as a
supplement to or replacement for the natural gas infrastructure, current production levels are insufficient to meet the
world's yearly demand. The question of what feedstock is most suited for the biogas economy is not easily answered. Proteins, lipids, and carbohydrates all have several potential uses. I.
INTRODUCTION A total of around 40 GWe of biomass-
produced electricity production capabilities (generating 160 TWh annually) and over 200 GW of heat generation 1 ISSN: 2959-832X Journal of Computational Intelligence in Materials Science 2(2024) capabilities (generating >700 TWh annually) were built across the globe by the expiration of the 19th century [2]. It is
projected that over 18 billion liters of biofuels were generated worldwide in 2016 (mostly ethanol made sugarcane and
cereals and maize surpluses, and to a much smaller level bio-fuels produced from the oil seed crop). This amounts to
around 0.5 EJ as transportation fuels (approximately 2000), although output of bio-ethanol across the globe is increasing
quickly. Bio-energy has the (technical) potential to make a significant contribution to the future global energy supply. With
expected technical advancements, energy cultivation on present farming land might theoretically make significant
contribution more than 800 EJ without threatening global food supplies. Another 40-170 EJ might come from organic
wastes and residues, with forest residues likely contributing the least and organic waste playing a potentially huge part,
particularly when biological materials are exploited on a grander scaling. The maximum potential for bioenergy may be
more than 1000 EJ (per annum) [3]. This is significantly more than the present world energy usage of approximately 400
EJ. Although Sub-Saharan Africa, Eastern Europe, and South America are the most immediately attractive locations,
Oceania and Northeast, and East Asia all stand out as prospective sites for biomass generation in the long-term future. The
latter is mostly explicable by demographic trends (China's population is expected to decline after 2020) and rapid technical
advancement in agriculture, which has resulted in significant productivity gains. These studies also reveal that substantial
portions of the biomass industry's technological potential may be realized at moderate production costs, on the order of 2
US dollars per gigajoule (GJ). However, significant changes are needed to fully tap into its bio-energy potential. Increasing
crop outputs per hectare is one measure of agricultural efficiency that is particularly important in developing nations. To
what degree and how quickly such transformations may be implemented in various places remains unknown. The (area)
bio-energy potentials could be significantly lower if circumstances are not optimal. It is important to highlight that technology advancements (in conversions, including long-term supply chains of
biomass (i.e.., global transportation of biomass produced energy transporters) may greatly increase the efficiency and
competitiveness of bio-energy. I.
INTRODUCTION One obvious motivator to explore the production possibilities of bio-energy is the need to
increase competitiveness. This paper provides a critical review of the current status of biomass conversion advancements
that are in use and innovations, which have a significant role to play in the near future, such as those linked to CO2
Collection and Sequestration (CCS). The generation of biodiesel for the transport sector is dignified to become the most
promising new marketplace for the widespread adoption of sustainable biomass practices. The rest of the paper is
organized as follows: Section II focuses on defining the technological background of biomass conversion. Section III
provides a critical discussion of combustion, bio-oil extraction, gasification, and digestion for heat and power. Section IV
is about the production of transport fuels through the method of gasification. Section V concludes the paper by providing
final remarks Domestic Heating Domestic Heating Producing heat for domestic usage is an ancient application of biomass combustion [6]. Domestic heating of biomass is
still a fundamental market for biomass in a significant portion of countries such as Germany, France, Sweden and Austria. In most countries, there is no hard data on how much wood is actually burned in fireplaces and stoves, although it is likely
that these sources contribute significantly to meeting the heat demand in the nations where it is mentioned. Using wood in
its traditional form is inefficient (as low as 10%) and produces large amounts of pollutants, such as dust and soot. Heating
systems have come a long way thanks to technological advancements, with many now being automated, including catalytic
gas purification, and using conventional fuels (e.g., pellets). The effectiveness value over the open fireplace is vital;
whereas the open fireplace could have a negative efficiency over the entire year (considering the loss of heat within the
chimney), modernized heaters could reach effectiveness of approximately 70% to 90% with significantly reduced
emissions. Such methods have found extensive use in nations such as Germany, Sweden and Austria. In Sweden, for
instance, the market for biomass pellets has grown rapidly due to the widespread use of automated firing systems. II.
TECHNOLOGY BACKGROUND OF BIOMASS CONVERSION Alternative fuels derived from agro-waste and biowaste
have attracted the attention of researchers due to the pressing need for environmentally responsible waste disposal on a
worldwide scale. There is a plethora of potential pathways for converting biomass into energy carriers. The primary conversion paths
utilized or being developed for the production of power, heat and transportation fuels are shown in Fig 1. In Section III, we 2 Journal of Computational Intelligence in Materials Science 2(2024) ISSN: 2959-832X will review the available or developing conversion advancements for generating heat and power (gasification, combustion,
and digestion), and in Section IV, we will do the same for transport biofuels (gasification, fermentation, and extraction),
differentiating between the two in terms of their potential and current availability. Fig 1. Conventional Methods for Biomass Conversion into Alternative Energy Sources
Combustions
Gasifications
HTU pryolytic
liquefaction
Digestion
Fermentations
Extractions
Heat
Electricity
Fuel
Steams
Turbines
Gases
Engine, combined
cycle and gas
turbines
Hydrogen,
hydrocarbon,
methanol synthesis
Fuel cells
Gases, Oils, and Char
Up-scaling
Diesel
Biogas
Engines
Distillations
Ethanol
Esterification
Diesel
Thermochemical conversions
Biochemical conversions Electricity Heat Heat Fig 1. Conventional Methods for Biomass Conversion into Alternative Energy Sources ds on several categories, which reflect a broad variety of technology ideas and deployment capability Fig 1 expands on several categories, which reflect a broad variety of technology ideas and deployment capability
ranges. III. COMBUSTION, BIO-OIL EXTRACTION, GASIFICATION, AND DIGESTION FOR HEAT AND POWER
Different alternatives exist for producing heat (industrial and domestic), Combined Heat and Power (CHP), and
electricity. In this section, we will examine the most prominent technologies that are now in use or have been used so far,
analyzing their function, current state, and baseline performance. Key choices now under development that may be pivotal
in the next decades are also highlighted. There is a focus on the European setting since so many cutting-edge bio-energy
technologies have been developed and implemented there. The technologies that may help nations in development with
low incomes are very briefly discussed. Combustion Larger-scale Combustion of Biomass g
f
Burning biomass is the simplest definition of biomass combustion. Humans have utilized this simple technology for
thousands of years to produce heat and, subsequently, to produce electricity via steam. Wood may be the most frequent
feedstock, but many other substances may also be burnt efficiently. Straw, sawdust, bark residuals, and shavings from
sawmills are all examples of byproducts and scraps that may be used as feedstock, as can "energy crops" like switchgrass,
poplar, and willow that are planted for that purpose. The pelletization of agricultural and wood leftovers is gaining
popularity as a convenient alternative. To save money, many rural residents, including farmers, are turning to biomass
heating systems instead of propane or heating oil. Direct space heating from a stove or fireplace is possible, or a back
boiler may be connected to provide hot water to radiators throughout the property. Pellet stoves are a relatively new
invention that utilizes an electrically powered auger to continuously feed compressed pellets of wood or other biomass into
the fire. It is possible to leave these stoves running for at least twenty-four hours without checking on them. Boilers fueled
by biomass may be used on a bigger scale to provide hot water, heat a building, or produce steam to operate machinery. Several farmers now rely on them as their only greenhouse heating source. Combustion of biomass at industrial scales is used all over the world to generate power (along with heat and process
steam). Throughout time, several variations on plant layouts have been created and put into use. The fundamentals of
combustion include pile burning, grate firing (vibrating, stationary, and moving), fluidized bed, and suspension firing
ideas. The Paper and Pulp (P&P) industry has been a pioneer in the usage of biomass combustion for power generation via
the burning of black liquor and waste incineration. For the most part, the P&P industry uses the conventional boiler for the
generation of electricity and steams as well as the recovery of pulping chemicals. From 1980s, countries such as the
Netherlands and Germany began widespread deployment of waste incinera tors with extremely tight emission rules. Such
facilities typically feature low stream temperatures and pressure (to deal with corrosion), high degrees of flue gas
purification, and enormous capabilities (i.e., approximately 1 Mtonne/plant/year). Larger-scale Combustion of Biomass Current electrical efficiencies range from the high teens to the low thirties, and more effective models (attaining about
30% power effectiveness) are being put into service. Mass burning has replaced landfills as the primary waste-to-energy
technique in Europe; however its high treatment costs of 50 to 150 Euros per tonne are offset by increased tipping costs. Independent plants for biomass combustion (by use of wood like forest left-overs as fuels) generally have capacities
between 20 MWe and 50 MWe, with the same power effectiveness in the range of 25% to 30%. Low prices of fuels, a
feed-in tariff, or a carbon tax for renewable power are necessary for such facilities to be profitable. Several cutting-edge
combustion ideas have been making their way into the mainstream in recent years. Producing both electricity (and heat)
and water from biomass is made possible by fluid bed technology and cutting-edge gas purification. Electrical efficiency
of 30% to 40% are feasible on a scale of around 50 MWe to 80 MWe. With their innovative CFB (Circulating Fluidized
Bed) and BFB (Bubbling Fluidized Bed) boilers, Finland is at the forefront of this sector. These boilers have great
flexibility of fuels, minimal investment expenses, high efficiency, and have been implemented on a massive scale. Finland's newest plant, with a 500 MWth capacity, is co-fired with different bio-fuels, some of which are delivered by
boat. District Heating and CHP Large-scale implementations of biomass-based district heating may be found in the Nordic nations and Austria. Because of
national climate and energy policy, biomass fuelled CHP models took off in the 1980s across Scandinavia. To begin, many
people opted to upgrade their already-existing coal-fired boilers. With the clear benefits of increased electrical efficiency
and reduced costs, there has been a rising trend in the size of CHP systems over time. This was coupled with the expansion
of the biomass market, which increased the accessibility and competitiveness of biomass resources sourced from greater 3 ISSN: 2959-832X Journal of Computational Intelligence in Materials Science 2(2024) distances (especially forest residues). Denmark launched a comprehensive plan to put straw to use in the 1990s. Cigar
burners and other effectual straw baling, storage, and transportation chains were among the many technological
innovations developed and put into practice. Straw's high alkali and chlorine content made traditional combustion methods
inefficient, therefore new approaches were required. These developments necessitated not just innovative pre-treatment
methods like straw washing, but also more sophisticated boiler ideas like two-stage combustion. While Austria is a global
leader in the deployment of biomass powered CHP, the majority of their systems are on the smaller scale of the village
level and are often integrated with local fuel delivery networks. All of these locations have colder winters than the United
States, making CHP a more viable option. In addition, it has been shown that community participation is crucial. Typically, CHP-plant owners are either local governments or forest owners. The energy expenses of such systems are
often greater. The local population backs the project very strongly, though, because of the jobs and money it brings into the
area. High labor expenses, however, have also resulted in extensive automation, with many modern plants operating
entirely without human oversight. ISSN: 2959-832X 1
Combustion
Chambers
Biomass
Bio-Char
Powder from
coal
Powder from
biomass
Air over-fire
Flue gases
Air
Air Biomass Mill No. 1 Torrefactions Combustion
Chambers Powder from
coal Bio-Char Mill No. 1 Fig 2. Torrefaction-Based Co-Firing Framework Co-firing Technologies
By burning both fossil fuels and biomass like coal or natural gas together in one power plant, known as "biomass co-
firing," more energy may be produced with less environmental impact. In the power industries, co-firing technologies may
be segmented into three distinct categories as shown in Table 1. Table 1. Categories of Co-Firing Technologies
Direct co-
firing
For existing coal-fired boilers, especially pulverized bio-coal boilers, direct co-firing is the most popular and
cost-effective way of co-firing biomass with coal. Dependent on the biomass's fuel properties and co-firing
ratio the bioenergy and coal are burnt together in the coal boilers furnace using the same or different mills Co firing Technologies
By burning both fossil fuels and biomass like coal or natural gas together in one power plant, known as "biomass co-
firing," more energy may be produced with less environmental impact. In the power industries, co-firing technologies may
be segmented into three distinct categories as shown in Table 1. Co firing Technologies
By burning both fossil fuels and biomass like coal or natural gas together in one power plant, known as "biomass co-
firing," more energy may be produced with less environmental impact. In the power industries, co-firing technologies may
be segmented into three distinct categories as shown in Table 1. Table 1. Categories of Co-Firing Technologies
Direct co-
firing
For existing coal-fired boilers, especially pulverized bio-coal boilers, direct co-firing is the most popular and
cost-effective way of co-firing biomass with coal. Dependent on the biomass's fuel properties and co-firing
ratio, the bioenergy and coal are burnt together in the coal boilers furnace using the same or different mills
and burners. Indirect
co-firing
Gasification, pyrolyzation or torrefaction of biomass produces upgraded solid biomass, gas fuel, and liquid
fuel, which are then co-fired in the furnace. Hence, there are three types of co-firing systems: those based on
torrefaction (which is what this thesis study focuses on), those based on gasification, and those based on
pyrolysis. Parallel
co-firing
To increase the conversion efficiency of the coal-fired energy plant, a biomass boiler may be installed beside
the existing boiler in a parallel co-firing arrangement. Table 1. ISSN: 2959-832X Journal of Computational Intelligence in Materials Science 2(2024) As many plants already have at least some co-firing capability, there is growing demand for even greater co-firing
shares (e.g., 40%). Nevertheless, there are more serious technical ramifications, such as for feeding lines and boiler
performance, and these are the ones that are now being developed to address. To take advantage of economies of scale and
decrease fuel supply threats, Denmark is constructing power plants (like the Avedore plant) that can combine natural gas
or coal with several types of biomass streams. Straw is widely used as a fuel source in Denmark. Problems with corrosion
and slagging in traditional combustion systems were brought on by the alkaline and chlorine rich straw. Straw, however,
may be used in multi-fuel systems to generate low-temperature steam, which can then be superheated using fossil fuels. These issues are mitigated to a large extent by using this method. These issues are mitigated to a large extent by using this method. Fig 2. Torrefaction-Based Co-Firing Framework
Co-firing Technologies
By burning both fossil fuels and biomass like coal or natural gas together in one power plant, known as "biomass co-
firing," more energy may be produced with less environmental impact. In the power industries, co-firing technologies may
be segmented into three distinct categories as shown in Table 1. Table 1. Categories of Co-Firing Technologies
Direct co-
firing
For existing coal-fired boilers, especially pulverized bio-coal boilers, direct co-firing is the most popular and
cost-effective way of co-firing biomass with coal. Dependent on the biomass's fuel properties and co-firing
ratio, the bioenergy and coal are burnt together in the coal boilers furnace using the same or different mills
and burners. Indirect
co-firing
Gasification, pyrolyzation or torrefaction of biomass produces upgraded solid biomass, gas fuel, and liquid
fuel, which are then co-fired in the furnace. Hence, there are three types of co-firing systems: those based on
torrefaction (which is what this thesis study focuses on), those based on gasification, and those based on
pyrolysis. Parallel
co-firing
To increase the conversion efficiency of the coal-fired energy plant, a biomass boiler may be installed beside
the existing boiler in a parallel co-firing arrangement. Torrefactions
Mill No. 1
Mill No. 1
Combustion
Chambers
Biomass
Bio-Char
Powder from
coal
Powder from
biomass
Air over-fire
Flue gases
Air
Air Fig 2. Torrefaction-Based Co-Firing Framework
Torrefactions
Mill No. 1
Mill No. Co-combustion In several European Union (EU) nations (such as the Netherlands, Spain, and Germany), biomass co-combustion, mostly
in coal-generated energy facilities, is the single most drastically advancing options of conversion for biomass. Co-benefits
firings are easy to see: when fuels of high quality such as pellets are utilized, the investment costs are cheap or nonexistent,
and the total electrical effectiveness is high (typically about 40%) [7]. Also, direct substitution of coal results in a
significant amount of reduced emissions. Moreover, many currently operating coal-fired power facilities have reached full
depreciation, making co-firing a particularly advantageous GHG mitigation alternative. Biomass fire also reduces
emissions of sulphur dioxide and other gases. Nonetheless, in certain nations, switching from coal to biomass might save
money on costly flue gas cleaning infrastructure upgrades. As a rule, boiler performance and upkeep are unaffected by low
co-firing shares. 4 ISSN: 2959-832X Categories of Co-Firing Technologies Direct co-firing has been the most common alternative for industrial biomass applications because of the cheap
investment cost of converting an existing coal power plant into a co-firing plant; nevertheless, high volatile content, low
density, and the high water content of biomass complicate its combustion behavior, restricting its co-firing ratios. Since it
may utilize coal, oil, or natural gas as its base fuel, indirect co-firing provides a great deal of fuel flexibility. Since
specialized biomass burners may be operated in parallel with the conventional boiler unit, the degree of business risks is
lowered and dependability is improved with parallel co-firing. Co-firing Solution for 100% Fuel Switching
Biomass is distinct from coal when it comes to combustion. Biomass loses a substantially larger percentage of its mass via
devolatilization than coal does; but, if the biomass nanoparticles are too big or thick, they may not be pressurized into the Co-firing Solution for 100% Fuel Switching Biomass is distinct from coal when it comes to combustion. Biomass loses a substantially larger percentage of its mass via
devolatilization than coal does; but, if the biomass nanoparticles are too big or thick, they may not be pressurized into the 5 Concept of Torrefaction based Co-firing System p
f
f
f
g y
Torrefaction greatly improves biomass's use and combustibility in combustion chambers (see Fig 2). One such pre-
treatment method that has showed promise and potential in recent years is torrefaction, which may be used to improve the
quality of biomass. Thermo-chemical fermentation process occurs between 200 and 300 ˚C, with a standard residence time
of 1 hour; during this time, biomass partly decomposes, releasing volatiles and generating the solid end products. During
thermal decomposition, the hemicellulose matrix that holds the raw biomass's cellulose fibers together is broken down,
resulting in shorter fibers and a loss of the raw biomass's tenacity. First, the weight and energy content of torrefied biomass
usually remain constant at 70% and 90%, respectively, compared to the original. In addition, the raw sample is both
challenging and energy-intensive to ground due to the higher fiber content of raw biomass. To assist the insertion of wood
powder into boiler furnaces, torrefied woody particles have a greater flowability and superior fluidization behavior than
raw biomass. Lastly, the quality of torrefied biomass is consistent between batches. After torrefaction, the physical and
chemical qualities of woodcuttings, destruction wood, and woody biomass are all the same, allowing for a wider range of
fuel options and mitigating the effects of seasonality. ISSN: 2959-832X Journal of Computational Intelligence in Materials Science 2(2024) flue gas, instead entering the bottom ash stream and undergoing minimal conversion beyond dehydration if the biomass
injection is not properly coordinated. Moreover, the biomass particles' low densities allow for oxidation at rates far greater
than those for coal. Moreover, striated flows are prevalent during biomass-coal combustion if the boilers fail to adequately
mix the flue gases in the furnace Chamber. While it is theoretically possible to co-fire with 20% biomass depending on the
energy content, most plants are only co-firing at 5- 10% levels at now. In most parts of the globe, pulverized fuel plants
have the highest installed capacity for burning coal. As a result, pulverized coal plants are the best candidate for co-firing. Hence, to achieve high levels of bioenergy co-firing in current pulverized coal boilers, a reliable biofuel co-firing solution
is necessary. Small-Scale Gasification Small-Scale Gasification
Small-scale gasification was heavily supported in the 1980s and 1990s. Small scaled power and power production using
gas or diesel engines led to the creation and testing of downdraft and updraft, fixed-bed gasifier with capabilities ranging
from under 100 kWth to several MWth. Commercial uses of tiny gasifiers for heat generation is well-established. Small-
scale (Bioneer) gasifiers for heating were widely used in Finland in the 1980s with great success. Nonetheless, combustion
is a formidable rival to gasification as a heat source. Using agricultural wastes near to their point of production has been a
central idea for quite some time. Several different gasifier, gas cleaning, and system integration ideas were developed and
put to the test under various situations. Under the help of organizations like the World Bank, technology was also sold to
other underdeveloped nations. Both rural electrification and economic growth were major factors. Small-scale gasification integrated with diesel or gas engines has not taken off despite significant research and
development efforts, financial expenditures, and a larger number of demonstration components. It is possible to purchase
small (fixed beds) gasifiers, which are integrated to gas/diesel engines (typically for 100 kWe to 200 kWe models with an
estimated, moderate, electrical effectiveness of 15% to 25%). Implementation has been successful, especially in India. Nevertheless, their widespread implementation in the EU has been delayed by the critical necessity of small-scale
glasifiers based on the quality of fuels (ideally conventional and hence more expensive fuels such as pellets) and cautious
operations, as well as high prices, notably for adequate gas cleaning given the stringent emission limits. Standardized
gasification models that are pre-packaged using micro-turbines and hydrogen fuels might be a game-changer for small-
scale power production from biomass in the long run, but such systems need further research and development, as well as
inexpensive and dependable fuel cells and, significant breakthroughs in small-scale gas cleansing. Gasification f
In the 1980s, both the United States and Europe paid a lot of attention to gasification as an approach to convert a wide
variety of solid bio-fuels into combustible syngas or gas [8]. When biomass has been gasified, the fuel gas may be further
processed or purified before being burned (for example, in gas turbines; whenever integrated with collective cycles, this
amounts in Biomass Integrated Gasification/ Combined Cycle (BIG/CC) plant). For further information on gasification's
role in the fuels industry, see Section IV. In the section, biomass gasification for the generation of heat and electricity will
be discussed first. We shall differentiate between gasification on a lesser scale (in several kWth to approximately 1 MWth
range of capacity and typically incorporating fixed-bed gasifications ideology) and large-scale gasification (often
associated with Fluid Bed perceptions). ISSN: 2959-832X ISSN: 2959-832X Journal of Computational Intelligence in Materials Science 2(2024) BIG/CC was the focus of several European Union (EU) and state projects. Research and demonstration projects have been
launched because of the potential of this technology to achieve great electrical efficiency at small scales with minimal
capital expenditures. In addition, the fuel gas must be rigorously cleaned before burning in order to fulfill gas turbine
criteria, hence BIG/CC ideas may reach minimal emission to air levels. BIG/CC was the focus of several European Union (EU) and state projects. Research and demonstration projects have been
launched because of the potential of this technology to achieve great electrical efficiency at small scales with minimal
capital expenditures. In addition, the fuel gas must be rigorously cleaned before burning in order to fulfill gas turbine
criteria, hence BIG/CC ideas may reach minimal emission to air levels. Some nations and gasification approaches have seen demonstration projects get off the ground. For example, in Brazil,
a World Bank/GEF financed initiative was established to showcase an ACFB BIG/CC unit with 30 MWe fueled with
grown Eucalyptus. During the same space of time, the first Swedish BIG/CC units (BIOFLOW piloted project) has logged
thousands of hours of operation using a pressurized gasification process. In 2000, the Yorkshire, UK facility that had been
testing an atmospheric BIG/CC cataloging switched to using natural gas instead of bio-fuels. The implementation of the
indirect FERCO concept of gasification in power plant in Burlington is a noteworthy project in the United States. There
have also been several national projects developed (especially in the EU) to create pre-commercial or demonstrator units
utilizing BIG/CC technologies. Nonetheless, putting the demonstration projects into action proved challenging. First
generation units were relatively expensive. Unit capital expenses are substantial for the initial generation of BIG/CC systems. Prices of between €5000 and
€3,500/kWe are quoted, which is still much over the target range of €1500 to €2000/kWe that would put BIG/CC in a
competitive position. There are still a number of technical challenges to be overcome, such as those associated with pre-
treatment and tar removal. Rapid market liberalisation in the energy industry in the late 1990s made it difficult for many
utilities to undertake costly demonstration programs. Recently, several of the demonstration units were decommissioned. The energy industry, which is notoriously risk adverse, tends to favor co-firing and established combustion technology
(which, of course, continues to advance over time). ISSN: 2959-832X As a result, progress on a technology that, in the not-too-distant future,
will be able to generate electricity from biomass at market-competitive prices, has stagnated. Many studies have analyzed
the cost-efficient potentials of the BIG/CC model, and at considerably higher scale (above 100 MWe) and according to the
continual advancement of gas turbine technologies, it is substantial. Use of grown biomass as fuel may be economically
viable in many parts of the globe due to the confluence of high electrical effectiveness and relatively cheaper unit capital
expenses. But, its progress has been modest thus far. Larger-scale Biomass Gasification g
f
Gasifiers with a capacity of many tens of megawatts of thermal energy (MWth) are often connected with Circulating
Fluidized Bed conceptions that provide critical fuel efficiency. It is true that gasifiers operating at ambient conditions
(ACFB) are employed in certain countries (such Italy, Austria, Sweden, and Germany) to generate (raw) producer gases
and process heat, but this is not a widespread practice. Biomass Power generation from Integrated Gasification/Combined
Cycle (BIG/CC) systems is very efficient and adaptable with regard to fuel characteristics [9]. Around 30 MWe is the size
at which 40% electrical efficiency (LHV basis) is achievable in the near future. Throughout the early part of the 1990s, 6 Gasification for Co-firing Larger co-firing conventional ratios in power stations may also be achieved by gasification, which eliminates the necessity
for additional solid fuel inputs and enhances control over the combustor. Recent evidence from co-firing systems
demonstrates the viability of CFB gasifier deployment (e.g. Amer in the Netherlands). Gasifier from gasification might be
used as a co-fuel in already-existing or newly-built combined cycles that utilize natural gas as their primary fuel source. Using economies of scale, one may achieve both relatively higher efficiencies and cheaper costs (presently up-to 60% for
NG fired cogeneration) while still having a reliable fuel supply, since the proportion of fuel gas to natural gas burnt can be
adjusted. While this has not yet been proved anywhere around the world, there has been an increase in study into it, and it
may prove to be of critical relevance in the near future as co-firing possibilities at unambiguous coal-fired energy plants
are progressively exploited. Bio-Oil Extraction Ablation involves applying pressure to a substance while moving it
rapidly over a heated surface. Using this novel quick heating method, big pieces of biomass may be heated directly without
being crushed beforehand. Further cracking processes, repolymerization, and bio-oil formation may be avoided if the
pyrolytic volatiles are quickly evacuated from the reactors and condensed, mostly in cooling systems. To create useful
reactive intermediates, a vacuum pyrolysis technology may be used instead of a carrier gas. The features of the biomass
and the operation settings determine the comparative yield and attributes of the liquid product. Biochemicals and biofuels
may be made from the liquid bio-oil that is produced when biomass is pyrolyzed. The impact of pyrolysis contexts of solid
products and the physio chemical characteristics of the chars have been studied in a number of publications that pyrolyzed
tobacco wastes in order to establish high-value products such as biochar. method is a thermal reactor used to generate heat. Ablation involves applying pressure to a substance while moving it
rapidly over a heated surface. Using this novel quick heating method, big pieces of biomass may be heated directly without
being crushed beforehand. Further cracking processes, repolymerization, and bio-oil formation may be avoided if the
pyrolytic volatiles are quickly evacuated from the reactors and condensed, mostly in cooling systems. To create useful
reactive intermediates, a vacuum pyrolysis technology may be used instead of a carrier gas. The features of the biomass
and the operation settings determine the comparative yield and attributes of the liquid product. Biochemicals and biofuels
may be made from the liquid bio-oil that is produced when biomass is pyrolyzed. The impact of pyrolysis contexts of solid
products and the physio chemical characteristics of the chars have been studied in a number of publications that pyrolyzed
tobacco wastes in order to establish high-value products such as biochar. Using TG-Fourier transform infrared (FTIR) and thermogravimetric (TG) analysis spectroscopy methods, some
researchers e.g., Ramsay [10] has studied the influence of temperatures and heating frequency on the products, uncovering
the possibilities of pyrolysis for the valorization of tobacco. In contrast to quick pyrolysis, which always results in a high
output of liquid oil, slow pyrolysis is known to produce a high yield of char. The yields and biochemical composition of
slow and quick pyrolysis were studied by Zaoui et al. [11]. Digestion
Biogas A wide range of feedstocks, including organic household waste, organic industrial by-products, manure sludges, etc., have
been shown to be suitable for anaerobic biomass digestion, and this method has been successfully utilized in commercial
settings. Whenever a digested gas is employed to energy gas engine-based generators, the total electrical efficiency is poor
(about 10-15%). Digestion is best for wet biomass sources. It is estimated that the maximum efficiency of converting
biomass into gas depends on the feedstock, but it is about 35%. In order to break down the large amounts of organic
material found in their wastewater, the food and beverage sector has traditionally relied on the process of digestion. Co-
digestion of manures as well as other wet organic wastes, for example, is proving to be especially successful in today's
market for sophisticated, large-scale models for wet industrial wastes. In the field of wet waste processing, nations such as
Denmark and Germany stand out because to their sophisticated digesting systems. Bio-Oil Extraction They discovered that the operating parameters were just as
important as the biomass type when it came to determining the yields and featuring the liquid oils. Wijayanti, Musyaroh, and Sasongko [12], the best liquid oil yields were generated at 600 ˚C. This was determined by
studying the influence of higher temperatures vacillating from 500 ˚C to 700 ˚C. The dispersal of chemical components in
the pyrolytic liquid oil was studied by Ourak et al., [13], who looked at the impact of temperatures (400 ˚C to 700 ˚C) and
additions (ZnCl2 and MgCl2). They discovered that the liquid oil could be converted into an easily ignitable fuel. Oil from
pyrolysis procedures and the resulting product are typically analyzed for chemical components using a 1D gas GC-MS
(chromatography-mass spectrometry). Modifying the procedure for py-GC-MS procedure, the TG-MS, and the TG-FTIR
might provide more understanding of chemical substances. g
p
g
Tobacco leaves and stems were analyzed for their chemical constituents by Sun and Sun [14]. They found that
secondary breakdown occurred at temperatures over 350 ˚C, resulting in the fragmentation of aromatic components into
smaller molecules. Nitrogen-containing compounds, phenols and nicotine are only some of the other chemical substances
that have been investigated. While one-dimensional GC has been in use for quite some time, it still has some minor
limitations when it comes to achieving the greatest possible compound separation. Higher-dimensional GC methods might
improve the detectability and resolution of liquid oils, enabling the identification of more chemicals. Many organic
molecules with the organic fractions may be identified using 2D, TOF/GCxGC MS evaluation as opposed to standard GC-
MS. Despite the abundance of research on 2D GC evaluation of bio-fuels and tobacco bio-oils is still uncommon. Bio-Oil Extraction Pyrolysis and Liquefaction Processes y
y
q
f
Biomass may be pyrolyzed at 500 ˚C temperature in the deficiency of oxygen to generate bio-oils, biogas and bio-char. The liquid fraction (70% or more of the biomass used for heating) may be optimized by the use of flash pyrolysis methods
(rapid pyrolysis). Oxygen makes up around 40% of bio-weight; oil's making it caustic and acidic. Engines and turbines
might theoretically be fueled by crude bio-oil (with some tweaks and only for higher grade oils). Upgrading the oil
(through hydrogenation) may also lower the oil's oxygen level. Nonetheless, there are costs associated with upgrades in
terms of money and energy. The majority of upgrading and pyrolysis technologies is still in the stage of demonstration. Other methods for creating 'raw intermediate' liquids from biomass include liquefaction (pressured conversion) and HTU
(Hydrothermal Upgrading) that convert high pressured biomass in moderate temperatures and water to bio-fuel (technique
in its early stages, developed first by Shell). In comparison to gasification, pyrolysis has seen less development thus far (and liquefaction options even more so). Potential small-scale technological deployment in rural surroundings and as feed-stock for the chemical sector has
attracted a lot of interest since the late 1980s and early 1990s. It was also argued that bio-higher oil's energy density
compared to untreated biomass would reduce transportation costs. While a lot of knowledge has been amassed over the
years, only a few of demonstration units have been fully implemented (Fortum, a Finnish oil firm, and Dynamotive, a
Canadian company at the forefront of commercializing pyrolysis technology, are excellent case studies). Market
implementation is still in its early stages. More and more people are looking at pyrolysis as a way to prepare bio-oil for
long-distance transport and subsequent conversion (refers to a wide range of applications, from (entrained flow)
gasification for syngas biosynthesis to efficient energy production). The term "pyrolysis" refers to a thermal cracking process used for biomass feedstock that either does not contain
oxygen or contains just a small amount of oxygen. In normal use, it is subjected to temperatures between 300 and 700 ˚C. The process of pyrolysis amounts to three major products: gas, solid char, and liquid oil. An essential component of the 7 ISSN: 2959-832X Journal of Computational Intelligence in Materials Science 2(2024) method is a thermal reactor used to generate heat. Landfill Gas Utilization f
The microorganisms living in a landfill produce a wide variety of gases known as landfill gas (LFG). Landfills that are just
not properly managed may release large amounts of LFGs like CH4 and CO2 into the atmosphere, causing significant
climate change; emit unpleasant odors, trash, and dust into the surrounding area; and allow leachate generated in the
landfill to leak into nearby ground and surface water. At 17.7% of the total CH4 released by the United States in 2011,
1908 landfills were responsible for producing roughly 1.03±108 metric tons of carbon equivalent of CH4. China has 580
landfills in 2013; 13 percent of the country's total CH4 emissions came from these sites. Estimated CH4 emissions from
waste disposal facilities in Europe made about 22% of the total, making them the second greatest source of CH4 generated
from human activity. If LFG emissions are allowed to continue unchecked, they will have the greatest impact on the
continent of Africa. Managing the created LFG is crucial since the total potential methane from Africa in 2012 was
10,496106 m3 (assuming all wastegenerated is landfilled). (
g
g
)
The term "LFG utilization" refers to the process of gathering and processing LFG obtained from the breakdown of
solidwaste from a landfill for the purpose of creating power, fuel, heat, and different valuable chemical compounds. An
expanding suite of LFG utilization technologies is now being put to use in a variety of contexts. Africa is home to just
around 1% of the world's LFG utilisation technology, and the continent is seeing a relatively slow pace of growth in this
area over the years. Africa is home to around 15% of the population within the globe, or more than 1.2 billion individuals. However, Africa relies heavily on landfilling for its Waste disposal. Hence, as MSW output is anticipated to rise as 8 Methanol, Hydrocarbons and Hydrogen via Gasification Liquid-phase methanol production, Fischer-Tropsch synthesizing (integrated with energy production), and new gas
separation and cleansing technologies are all examples of cutting-edge technical ideas that might lead to reduced
production costs and greater overall efficiency in the long run. Nevertheless, further Research and development (R&D)
efforts over a significant duration of time are needed to accomplish such as state of affairs. The efforts to enhance
gasification technologies for the production of syngas are on the increase in nations like Sweden, the Netherlands, and
Germany. Energy efficiency in somewhat "standard" manufacturing plants may approach 60% (on a 400 MWth input
scale). Deployment, in large-scale, (e.g., above 1000 MWth) is necessary to fully take advantage of the economies-of-scale
intrinsic to these kinds of projects. From an economic and efficiency standpoint, however, this choice (or group of options)
stands out as a clear winner. By gasifying biomass, we may get methanol, hydrogen, and Fischer-Tropsch diesel. Prior to producing the secondary
power carrier by relatively typical gas processing technologies, all approaches need very pure syngas. SNG (Synthetic
Natural Gas), and DME (DiMethylEther) could be established from syngas in addition to MeOH, hydrogen, and FT-
liquids. Many approaches using either already available technology or cutting-edge, experimental ones are feasible. A
typical conversion flowchart for such procedures is shown in Fig 3. Before the formaldehyde or FT reactor, or hydrogen
separation, there is a sequence of operations to convert the biomass to the desired gas requirements. p
q
p
g
q
Syngas, which includes carbon monoxide (CO), hydrogen (H2), and a few other chemicals, is what comes out of the
gasifier. There follows a chain of biochemical processes on the syngas. Except the gas cleaning train, the machinery
required to produce hydrogen gas, methanol, and fuel-grade diesel from natural gas is identical. Gas turbines, boilers, and
steam turbines may all be used to co-generate electricity from the remaining unconverted gas fractions. Even though there
hasn't been any commercial biofuels production using gasification yet, there has been a lot of research and development
into the process over the previous several decades. For the purpose of creating transportation fuels (like methanol), interest
in syngas made from biomass emerged in the 1980s, in part as a response to the escalating price of oil. French and Swedish
researchers have been working to perfect the process of pressured gasification for creating methanol from biomass. ISSN: 2959-832X Journal of Computational Intelligence in Materials Science 2(2024) Africa's population rises, an expansion in landfill sites is anticipated. The continent has not been able to make good use of
the LFG it generates from landfills, and neither African policymakers nor academics have paid much attention to the
process of enriching the LFG that is released from African landfills. Gases produced from landfills may provide viable
options for the supply of energy in several African nations. According to estimates, the conversion of LFG to electricity in
Kampala (Uganda) reached 31,000 MWh in 2009 and 26,600 MWh in 2011 [15]. It is being considered as a significant
source of power in South Africa, Nigeria, and other African nations. p
g
Landfills are one particular location from which biogas may be extracted. Landfill sites are a major source of methane
emissions since they produce methane-rich landfill gas. Profitability has led to the widespread implementation of systems
for collecting landfill gas and turning it into power using gas engines. It is easy to see the advantages: This process diverts
methane gas, which has increased global warming potentials compared to carbon dioxide (CO2), from landfills and
converts it into usable energy carriers. As a result, reusing landfill gas has become a popular strategy for reducing
greenhouse gas emissions in many parts of the globe, including the United States (US), European Union (EU), and
increasingly other parts of the world. IV.
PRODUCTION OF TRANSPORT FUELS THROUGH GASIFICATION Fig 1 shows the three primary pathways to generate transportation fuels from biomass. Gasification may provide
syngas, which can then be transformed into methanol, DiMethylEther (DME), Fischer-Tropsch liquids, and hydrogen. The
most common method for making biofuels involves hydrolysis procedures to convert ligno-cellulosic biomass into sugars
before the biomass is fermented into ethanol. Last but not least, oil seeds (such as rapeseed or palmoil) may be extracted
for their oil, which can then be esterified to create biodiesel for use as a biofuel. Despite its small weight, hydrogen needs a
complex infrastructure to transport and store. Except for DME, all of the other potential fuels are liquids that may be stored
and supplied using existing facilities. pp
g
g
As ethanol and methanol in particular have a lower power density than gasoline, more mass must be carried on board
to achieve the same level of energy output. The fuels' potential toxicity and disaster-related environmental effects are also
important considerations. Benzene and other carcinogenic aromates are present in gasoline and diesel. Although methanol
does not cause cancer, it is more harmful to human skin than gasoline. Closed filling systems and other precautions are
needed to make the fuel safer for consumers to handle than gasoline and diesel (e.g. as employed for LPG). Investment
expenses will go up as a consequence of this. As compared to traditional fuels like gasoline and diesel, Fischer-Tropsch
liquid and ethanol have significant benefits, including a negligible toxicity as well as a complete absence of sulphur and
aromates. As an added bonus, we can use the gasoline and diesel distribution networks already in place. ISSN: 2959-832X ISSN: 2959-832X has been a resurgence of interest in gasification technology for the manufacture of transportation fuels ever since e.g.,
Sharma, Antil, and Sachdeva in [16]. has been a resurgence of interest in gasification technology for the manufacture of transportation fuels ever since e.g.,
Sharma, Antil, and Sachdeva in [16]. has been a resurgence of interest in gasification technology for the manufacture of transportation fuels ever since e.g.,
Sharma, Antil, and Sachdeva in [16]. Fig 3. Using plant matter for production of fuels including hydrogen, methanol, and FT diesel by gasification
Biomass
Gas cleansing
and
gasification
Chipping and
drying
Separation,
shifting,
reforming,
CO2
Separation
Separation,
catalysis
Separation,
catalysis
Hydrogen
Methanol
Diesel
Power
Refinement
Boilers or gas
turbines
Steam
turbines Gas cleansing
and
gasification Biomass Power Steam
turbines ig 3. Using plant matter for production of fuels including hydrogen, methanol, and FT diesel by gasificatio ant matter for production of fuels including hydrogen, methanol, and FT diesel by gasification Choren, a German business, has shown how to make FT-diesel from biomass gasification in Freiburg. Even though the
technological and economic promise of such ideas is significant, their implementation seems feasible. However,
technological hurdles remain, and they are most probably becoming challenging compared to those facing BIG/CC
ideologies. This is due to the fact that more complex gas cleansing is required to safeguard downstream catalytic gaseous
production equipment. Once pure syngas is readily accessible, established methods for making FT-liquids, methanol,
hydrogen, and DME may be put into practice. Gas purification, process scaling, and integration into existing infrastructure
are the primary obstacles to progress. V.
CONCLUSION Throughout the 1990s and 1980s, bio-energy and biomass were mostly considered at the regional and local levels;
however, this is becoming a global issue. The internationalization of biomass markets has led to an increase in the export
of biomass and biomass-retrieved energy transferors. In addition, it becomes more challenging to keep extremely distinct
national policies due to carbon and certificate trading, including projects employed as under the Clean Growth Approach
or the Joint Operational Activities. An enthralling example of the pan-European objective with potentially significant
implications for a European bio-energy marketplace is the new EC biofuel regulation, which mandates biomass resource
generation and transportation of high-quality fuels. More competitive, sophisticated and efficient conversion capabilities,
particularly, for the production of heat and power, must be commercialized, and similar considerations apply to
technological advancements and the RDD&D trends required doing so. Biomass is one renewable power source that may
significantly contribute to the globe's power production in the future. Bioenergy's exact role will be determined by its
rivalry with fossil fuels and by agriculture policy across the globe, although it appears reasonable to predict a significant
rise from the present contribution of 40-55 EJ per year. With possible observations of 200–300 EJ lasting far beyond this
century, biomass has surpassed mineral oil as a viable energy supply source. Bio-energy has serious challenges, one of
which is the need for its usage to be updated so that it may be integrated into sustainable development strategies. Data Availability y
No data was used to support this study. Conflicts of Interests
The author(s) declare(s) that they have no conflicts of interest. Conflicts of Interests
The author(s) declare(s) that they have no conflicts of interest. Methanol, Hydrocarbons and Hydrogen via Gasification Large-scale CFB gasifiers were structured in Finland by fertilizer manufacturer Kemira to generate syngas for use in the
city's ammonia plant (which was shut down). Important industrial experience with this technique can be found at Schwarze
Pumpe (East Germany), where gasification capabilities (entrained flow) has been built to produce methanol from waste
streams. Advanced gasification techniques for widespread use are in a precarious position due to low energy costs. There 9 Journal of Computational Intelligence in Materials Science 2(2024) There are no competing interests. There are no competing interests. References eferences
[1]. A. Kumar Tripathi, I. Patra, N. Bharath Kumar, A. Majdi, I. Muda, and A. Mahdavi, “Environmental and exergoeconomic assessments of a
novel biomass gasification based solid oxide fuel cell and heat engine hybrid energy system,” Energy Sources Recovery Util. Environ. Eff.,
vol. 44, no. 4, pp. 8490–8511, 2022. pp
an, C. O’Dwyer, and D. Flynn, “Fast frequency response provision from commercial demand response, from scheduling to
wer systems,” IET Renew. Power Gener., vol. 16, no. 9, pp. 1908–1924, 2022. [2]. M. S. Misaghian, C. O’Dwyer, and D. Flynn, “Fast frequency response provision from commercial demand response
stability in power systems,” IET Renew. Power Gener., vol. 16, no. 9, pp. 1908–1924, 2022. y
p
y
pp
[3]. A. Iqbal, N. Noreen, M. Imran, J. L. Francisco Alves, J. C. G. da Silva, and S. L. Badshah, “Valorization of the biomass of Rhizoclonium
hookeri through slow pyrolysis and its thermokinetic investigation for bioenergy potential,” Biomass Bioenergy, vol. 168, no. 106690, p. 106690, 2023. [4]. P. Basinas, J. Rusín, K. Chamrádová, and S. P. Kaldis, “Pyrolysis of the anaerobic digestion solid by-product: Characterization of digestate
decomposition and screening of the biochar use as soil amendment and as additive in anaerobic digestion,” Energy Convers. Manag., vol. 277,
no. 116658, p. 116658, 2023. p
[5]. Z. Lajdova, J. Lajda, and P. Bielik, “The impact of the biogas industry on agricultural sector in Germany,” Agric. Econ., vol. 62, no. 1, pp. 1–
8, 2016. [6]. S. Caserini, S. Livio, M. Giugliano, M. Grosso, and L. Rigamonti, “LCA of domestic and centralized biomass combustion: The case of
Lombardy (Italy),” Biomass Bioenergy, vol. 34, no. 4, pp. 474–482, 2010. u, W. Tan, S. Liang, X. Bi, R. Sun, and X. Pan, “Comparative study on the co-combustion behavior of torrefied biomass blended w
ent rank coals,” Biomass Convers. Biorefin., 2022. different rank coals,” Biomass Convers. Biorefin., 2022. [8]. G. S, D. T, and A. Haldorai, “A Supervised Machine Learning Model for Tool Condition Monitoring in Smart Manufacturing,” Defence
Science Journal, vol. 72, no. 5, pp. 712–720, Nov. 2022, doi: 10.14429/dsj.72.17533. drigues, A. Walter, and A. Faaij, “Co-firing of natural gas and Biomass gas in biomass integrated gasification/combined cycle system
y (Oxf.), vol. 28, no. 11, pp. 1115–1131, 2003. msay, “Uncertainty quantification of an explicitly coupled multiphysics simulation of in-situ pyrolysis by radio frequency heating in
” SPE J., vol. 25, no. 03, pp. 1443–1461, 2020. pp
[11]. Funding No funding was received to assist with the preparation of this manuscript. 10 10 Journal of Computational Intelligence in Materials Science 2(2024) ISSN: 2959-832X ISSN: 2959-832X References N. Zaoui et al., “Assessment of effects of metal contamination and abiotic factors on fatty acid composition and biochemical biomarkers
activity in the liver of Chelon ramada collected from two ecosystems on the Mahdia coast, Tunisia,” Biochem. Syst. Ecol., vol. 107, no. 104608, p. 104608, 2023. , p
,
[12]. W. Wijayanti, M. Musyaroh, and M. N. Sasongko, “Low-density polyethylene plastic waste to liquid fuel using pyrolysis method: An effect of
temperatures on the oil yields physicochemical properties,” J. Sustain. Dev. Energy Water Environ. Syst., vol. 10, no. 3, pp. 1–18, 2022. [13]
M O
k
l “E
i
l
d
f
l i
il f
d i
I
f
d
i
li
id
i i
” W
Bi . M. Ourak et al., “Experimental study of pyrolytic oils from used tires: Impact of secondary reactions on liquid composition,” Wa
Valorization, vol. 12, no. 8, pp. 4663–4678, 2021. [14]. H.-R. Sun and M. Sun, “Study on chemical constituents from tobacco leaves,” DEStech Trans. Eng. Technol. Res., no. iceea, 2017. ]. A. N. Srivastava and S. Chakma, “Quantification of landfill gas generation and energy recovery estimation from the municipal
landfill sites of Delhi, India,” Energy Sources Recovery Util. Environ. Eff., pp. 1–14, 2020. [16]. A. K. Sharma, S. Antil, and G. Sachdeva, “Small-scale toroidal fluidized bed gasification system: Development and experimental aspects,” J. Energy Resour. Technol., vol. 145, no. 5, pp. 1–19, 2023. 11 11
|
https://openalex.org/W2912974090
|
https://europepmc.org/articles/pmc6449486?pdf=render
|
English
| null |
The first identification of nesfatin-1-expressing neurons in the human bed nucleus of the stria terminalis
|
Journal of neural transmission
| 2,019
|
cc-by
| 4,224
|
Journal of Neural Transmission (2019) 126:349–355
https://doi.org/10.1007/s00702-019-01984-3 Journal of Neural Transmission (2019) 126:349–355
https://doi.org/10.1007/s00702-019-01984-3 YCHIATRY AND PRECLINICAL PSYCHIATRIC STUDIES - SHORT COMMUNICATION Abstract Neuropeptides are involved in various brain activities being able to control a wide spectrum of higher mental functions. The purpose of this concise structural investigation was to detect the possible immunoreactivity of the novel multifunc-
tional neuropeptide nesfatin-1 within the human bed nucleus of the stria terminalis (BNST). The BNST is involved in the
mechanism of fear learning, integration of stress and reward circuits, and pathogenesis of addiction. Nesfatin-1-expressing
neurons were identified for the first time in several regions of the BNST using both immunohistochemical and fluorescent
methods. This may implicate a potential contribution of this neuropeptide to the BNST-related mechanisms of stress/reward
responses in the human brain. Keywords BNST · Nesfatin-1 · Brain · Neuropeptides Keywords BNST · Nesfatin-1 · Brain · Neuropeptides The first identification of nesfatin-1-expressing neurons in the human
bed nucleus of the stria terminalis Artur Pałasz1 · Katarzyna Bogus1 · Aleksandra Suszka‑Świtek1 · Andrzej Kaśkosz2 · Shirley Saint‑Remy3 ·
Aneta Piwowarczyk‑Nowak2 · Łukasz Filipczyk1 · John J. Worthington4 · Kinga Mordecka‑Chamera1 · Karol Kostro2 ·
Grzegorz Bajor2 · Ryszard Wiaderkiewicz1 Received: 8 December 2018 / Accepted: 6 February 2019 / Published online: 15 February 2019
© The Author(s) 2019 4
Division of Biomedical and Life Sciences, Faculty of Health
and Medicine, Lancaster University, Lancaster LA1 4YQ,
UK Introduction involved in the neuromechanism of addiction and feeding
behavior (Ch’ng et al. 2018; Avery et al. 2016; Pleil et al. 2016). The potential role of BNST-related circuits in the
origin of some psychopathic traits has also been postulated
(Schiltz et al. 2007). Both the human and animal BNST
are functionally connected with limbic structures, thalamic
nuclei, and basal ganglia, while newly reported connections
with temporal and paracingulate cortex are exclusive to
the human brain (Avery et al. 2014). The human BNST, a
relatively small (average volume ~ 180–190 mm3) droplet-
shaped neural association is located in the central forebrain
and subdivided along a medial–lateral axis consisting of four
areas: medial (BNSTM), central (BNSTC), lateral (BNSTL),
and ventral (BNSTV) (Theiss et al. 2017). Interestingly, the
BNSTC represents a distinct sexual dimorphism, being
larger in men than in women (Swaab 2007; Chung et al. 2002). The detailed neuropeptide profile of BNST has been
previously reviewed (Kash et al. 2015). The BNSTM has a
dense noradrenergic innervation (high β-hydroxylase immu-
noreactivity), the BNSTC contains an abundant population
of somatostatin (SOM) neurons, whereas the more hetero-
genic BNSTL is characterized by SOM, cholecystokinin
(CCK), NPY, and neurotensin expression (Walter et al. 1991;
Martin et al. 1991). On the other hand, oxytocin signaling
in the rat BNST seems to be involved in the mechanism of The bed nucleus of the stria terminalis (BNST), a part of
the so-called “extended amygdala”, is a minor gray mat-
ter aggregation located in the medial basal forebrain of
vertebrate species. Accumulating evidence prove that this
intriguing structure plays a crucial role in the integration of
stress and reward signaling, generation of anxiety responses,
and regulation of fear learning (Harris et al. 2018; Pelrine
et al. 2016; Rodriguez-Sierra et al. 2016). This maybe highly * Artur Pałasz
apalasz@sum.edu.pl 1
Department of Histology, School of Medicine in Katowice,
Medical University of Silesia, ul. Medyków 18,
40‑752 Katowice, Poland 2
Department of Anatomy, School of Medicine in Katowice,
Medical University of Silesia, ul. Medyków 18,
40‑752 Katowice, Poland 2
Department of Anatomy, School of Medicine in Katowice,
Medical University of Silesia, ul. Medyków 18,
40‑752 Katowice, Poland 3
American Medical Student Association (AMSA), School
of Medicine in Katowice, Medical University of Silesia,
ul. Medyków 18, 40‑752 Katowice, Poland 3
American Medical Student Association (AMSA), School
of Medicine in Katowice, Medical University of Silesia,
ul. 3
American Medical Student Association (AMSA), School
of Medicine in Katowice, Medical University of Silesia,
ul. Medyków 18, 40‑752 Katowice, Poland Introduction Medyków 18, 40‑752 Katowice, Poland 4
Division of Biomedical and Life Sciences, Faculty of Health
and Medicine, Lancaster University, Lancaster LA1 4YQ,
UK (0121 3456789)
3 350 A. Pałasz et al. social recognition in rats and microinjections of oxytocin
into these structures enhanced social memory in male, but
not female animals (Dumais et al. 2016). side of the anterior commissure, superior was delineated
by the most ventral edge of the caudate nucleus and lateral
ventricle. Column of the fornix and internal capsule formed
medial and lateral borderlines, respectively. The sections
were deparaffinized with xylene and rehydrated to the 50%
ethanol by successive changing of an alcohol gradient. Nesfatin-1 is a recently discovered NEFA/nucleobindin-2
(NUCB2)-derived multifunctional neuropeptide (Schalla
and Stengel 2018; Pałasz et al. 2012). Nesfatin-1 is known
as a potent anorexigenic factor, inducing satiety, and inhibit-
ing food and water intake (Wernecke et al. 2014; Stengel and
Tache 2013). Intriguingly, a number of recent studies dem-
onstrate that nesfatin-1 plays an important role in other auto-
nomic and mental functions such as sleep–wake regulation
(Vas et al. 2013; Jego et al. 2012), anxiety or stress-related
responses (Pałasz et al. 2018; Emmerzaal and Kozicz 2013;
Merali et al. 2008), and may also be involved in the patho-
genesis of some psychiatric disorders (Weibert et al. 2018;
Xu et al. 2018; Shimizu and Mori 2013; Gunay et al. 2012;
Ari et al. 2011). The rat hypothalamus, arcuate, paraven-
tricular, and supraoptic nuclei as well as in dorsomedial and
lateral hypothalamus are characterized by distinct expression
of nesfatin-1. In addition, perikarya of the piriform, insu-
lar and cingulate cortex, amygdala, BNST, lateral septum
and zona incerta also demonstrate nesfatin-1 immunoreac-
tivity (Goebel-Stengel and Wang 2013; Stengel and Tache
2010). Nesfatin-1 neurons were also identified in cerebellum
(Purkyne cells) and numerous brainstem structures including
solitary tract, raphe nuclei, gigantocellular reticular nucleus,
lateral parabrachial nucleus, nucleus ambiguous, and central
gray (nucleus O). Despite the accumulating animal studies
on nesfatin-1 its presence and action are so far understudied
in the human brain structures. The aim of the current his-
tological study was to detect the presumptive existence of
nesfatin-1-expressing neurons in the human BNST. A num-
ber of psychiatric disorders may potentially be connected
with impaired neuropeptide-dependent regulation in BNST,
hence this morphological study offers potential mechanistic
understanding of neuropsychiatric disease. Introduction After rehydration, and subsequent antigen retrieval with
citrate buffer (pH 4.0) solution (vector laboratories), sections
were rinsed three times for 5 min in 0.05 M TBS-saline (pH
7.6) and placed in the 0.1% buffered Triton X-100 (Sigma)
to improve the antibodies penetration and reaction quality by
reducing surface tension of aqueous solutions during immu-
nohistochemistry. They were blocked with 10% goat serum
and incubated overnight at 4 °C with a mouse monoclonal
antibody (I) against rat nesfatin-1 (1:1000, Enzo Life Sci-
ences, IF) or rabbit anti-rat nesfatin-1 antibody II (1:5000
Phoenix Pharmaceuticals, IHC-P/IF). We decided to apply
two antibodies both mono- and polyclonal comparatively
to improve the immunostaining reliability. Cross-reactivity
of the antibody II with the human cells was reported previ-
ously (Ramanjaneya et al. 2010). Sections were addition-
ally treated with TrueBlack® Lipofuscin Autofluorescence
Quencher (Biotium, Hayward, CA, USA) to minimize
unwanted intense lipofuscin signal. After incubation with
primary antibodies, brain sections were kept in darkness
with appropriate secondary antibodies: goat anti-mouse
and goat anti-rabbit labeled with TRITC or FITC (1:200,
Abcam), respectively, and mounted on slides with DAPI-
containing medium. Neurosecretory cells of the hypotha-
lamic supraoptic nuclei (SON) with a distinct intensity of the
fluorescent signal (a proven high level nesfatin-1 expression)
represent an internal positive control. Alternatively, primary
antibody I was followed by biotinylated rabbit anti-goat sec-
ondary antibody, and then an avidin-biotin-horseradish per-
oxidase complex (Vectastain ABC kit, Vector Labs). Finally,
3,3′-diaminobenzidine (DAB) was used to visualize the reac-
tion. All sections were mounted on glass slides, dehydrated,
and coverslipped. For basic neurostructural evaluation rep-
resentative sections were stained with Nissl method in 1%
of water solution of Cresyl violet for 60 min. After rinsing
and differentiation by acetic acid and mounting with DPX,
sections were preserved with cover glasses. All images were
captured on a Nikon Coolpix fluorescent optic systems and
processed using Image ProPlus software (Media Cybernet-
ics, USA). Despite the much extended process of tissue fixa-
tion, the general neuronal morphology was sufficient to per-
form immunohistochemical analysis. The BNST cyto- and
chemoarchitecture was analyzed and immunopositive cells
were counted using ImageJ 1.43u software. The anatomical
coordinates and structure of BNST were defined according
to Mai human brain atlas (Mai et al. 2015). Positive cells
were divided per area of the analyzed BNST subdivision;
medial (BSTM) ventral (BSTV) and lateral (BSTL) to Materials and methods Studies were carried out on human brain tissue specimens
with no neuropathological findings obtained within the Con-
scious Body Donation Program conducted by the Depart-
ment of Anatomy at the Medical University of Silesia in
Katowice. The brains were postmortem perfused and fixed
with buffered solution 4% formaldehyde (pH 7.2–7.4) over
a period of at least 3 months. The tissue samples containing
BNST were precisely excised from two forebrain specimens
according to Mai, Majtanik and Paxinos Atlas of the Human
Brain (2015), dehydrated, embedded in paraffin, and finally
sectioned on a microtome (Leica Microsystems, Germany)
at 10-µm-thick serial slices (Fig. 1). The inferior BNST
boundary was identified in the coronal plane by the superior 1 3 3 351
The first identification of nesfatin-1-expressing neurons in the human bed nucleus of the stria…
Fig. 1 Overview of the analyzed coronal serial sections of the human
brain pericommissural area at the level of BNST (red). Sections G
and H contain the hypothalamic supraoptic nucleus (SON, yellow);
anterior commissure, ac; lateral ventricle, lv; hypothalamus, hyp;
third ventricle, 3v. Nissl staining. Scale bar: 5 mm. Immunofluores-
cence for nesfatin-1 in the human BNST. Cell bodies of the BNSTM
(a–c) and BNSTC (d–f) subnuclei (TRITC). Neuropeptide immuno-
positive somata and fibers in the BNSTC (FITC, g). Magnocellular
perikarya of the hypothalamic supraoptic nuclei (SON, h, i) with a
distinct intensity of the fluorescent signal (a proven high level nes-
fatin-1 expression) represent an internal positive control. Scale bars:
200 µm and 100 µm (i, inset) The first identification of nesfatin-1-expressing neurons in the human bed nucleus of the stria… 351 Fig. 1 Overview of the analyzed coronal serial sections of the human
brain pericommissural area at the level of BNST (red). Sections G
and H contain the hypothalamic supraoptic nucleus (SON, yellow);
anterior commissure, ac; lateral ventricle, lv; hypothalamus, hyp;
third ventricle, 3v. Nissl staining. Scale bar: 5 mm. Immunofluores-
cence for nesfatin-1 in the human BNST. Cell bodies of the BNSTM (a–c) and BNSTC (d–f) subnuclei (TRITC). Neuropeptide immuno-
positive somata and fibers in the BNSTC (FITC, g). Magnocellular
perikarya of the hypothalamic supraoptic nuclei (SON, h, i) with a
distinct intensity of the fluorescent signal (a proven high level nes-
fatin-1 expression) represent an internal positive control. Scale bars:
200 µm and 100 µm (i, inset) 1 3 352
A. Pałasz et al. Fig. 2 Immunohistochemical reaction for nesfatin-1 in the human
BNST. Materials and methods General microstructure and cell diversity of the central
(BNST, c, a) and ventral (BNSTV, b, d) subdivision; Nissl staining. Neuropeptide immunopositive perikarya in the respective regions
(A1, A2, white arrows); DAB reaction. A selection of neurones from
the picture B1 under higher magnification (B2, B3). Immunopositive
fibers in the BNSTC (fluorescence, FITC, c), numerous cell bodies
contain deposits of lipofuscin with a strong yellow autofluorescence
(inset, blue arrows). Mean number of nesfatin-1 immunopositive cells
per standardized frame (0.2 mm2) located at the intersections of dot-
ted lines (a preliminary analysis). Values for two BNST subnuclei
taken from selected brain sections (see Fig. 1). Scale bars: 200 µm
(A, A1, B, B1, D), 100 µm (B2), 50 µm (B3) A. Pałasz et al. 352 Fig. 2 Immunohistochemical reaction for nesfatin-1 in the human
BNST. General microstructure and cell diversity of the central
(BNST, c, a) and ventral (BNSTV, b, d) subdivision; Nissl staining. Neuropeptide immunopositive perikarya in the respective regions
(A1, A2, white arrows); DAB reaction. A selection of neurones from
the picture B1 under higher magnification (B2, B3). Immunopositive
fibers in the BNSTC (fluorescence, FITC, c), numerous cell bodies
contain deposits of lipofuscin with a strong yellow autofluorescence
(inset, blue arrows). Mean number of nesfatin-1 immunopositive cells
per standardized frame (0.2 mm2) located at the intersections of dot-
ted lines (a preliminary analysis). Values for two BNST subnuclei
taken from selected brain sections (see Fig. 1). Scale bars: 200 µm
(A, A1, B, B1, D), 100 µm (B2), 50 µm (B3) Fig. 2 Immunohistochemical reaction for nesfatin-1 in the human
BNST. General microstructure and cell diversity of the central
(BNST, c, a) and ventral (BNSTV, b, d) subdivision; Nissl staining. Neuropeptide immunopositive perikarya in the respective regions
(A1, A2, white arrows); DAB reaction. A selection of neurones from
the picture B1 under higher magnification (B2, B3). Immunopositive
fibers in the BNSTC (fluorescence, FITC, c), numerous cell bodies contain deposits of lipofuscin with a strong yellow autofluorescence
(inset, blue arrows). Mean number of nesfatin-1 immunopositive cells
per standardized frame (0.2 mm2) located at the intersections of dot-
ted lines (a preliminary analysis). Values for two BNST subnuclei
taken from selected brain sections (see Fig. 1). Results and discussion i
Our results demonstrate for the first time the presence
of nesfatin-1 in the human limbic structures which could
implicate a potential contribution of this factor to the central
mechanisms of stress/reward responses. Currently, a role for
nesfatin-1 in human BNST physiology remains speculative,
but undoubtedly further studies on this novel regulatory fac-
tor, e.g., its coexpression with other important BNST neu-
ropeptides such as CRF, NPY, somatostatin and analysis of
potential functional relationship between these regulatory
factors, strongly merit attention. Moreover, the 3-D visuali-
zation of several novel multifunctional neuropeptides (nes-
fatin-1, phoenixin, spexin) spatial distribution in the human
BNST, amygdaloid complex, other limbic structures includ-
ing several basal ganglia and hypothalamus should be of
primary focus. We have demonstrated for the first time nesfatin-1 immu-
noreactive neurons in the human BNST which suggests
that this novel neuropeptide may be involved in functions
realized by this brain region. Numerous nesfatin-1-positive
neurons are present in the human BNST and their assemblies
show different patterns of distribution in selected BNST sub-
nuclei. Highest density of medium-sized (20–30 µm), nes-
fatin-1-immunoreactive cells were observed in the BNSTM
sections; mean number per standardized frame area was
200 ± 12%. In the BNSTC, a smaller proportion of multi-
form neuropeptide immunopositive cells was found depend-
ing on the section plane starting of more rostral orientation
(97 ± 16.4%, 40 ± 2% and 75 ± 10%, respectively, Figs. 1,
2). Of note, the relatively large (40–50 µm), oval-shaped
perikarya of the ventral BNSTC exhibits high nesfatin-1
immunoreactivity (Fig. 2; b1–b3). The chemoarchitecture
of nesfatin-1 neurons in the BNST seems to be similar to the
distribution of NPY (Adrian et al. 1983), PACAP (Palkovitz
et al. 1995), and dynorphin-A neurons (Poulin et al. 2009). Acknowledgements This work was supported by the Medical Uni-
versity of Silesia statute grant for Department of Histology no:
KNW-1-171/N/8/I. Open Access This article is distributed under the terms of the Crea-
tive Commons Attribution 4.0 International License (http://creativeco
mmons.org/licenses/by/4.0/), which permits unrestricted use, distribu-
tion, and reproduction in any medium, provided you give appropriate
credit to the original author(s) and the source, provide a link to the
Creative Commons license, and indicate if changes were made. et al. 1995), and dynorphin A neurons (Poulin et al. 2009). At present, there is no consensus in the role of nes-
fatin-1 to the function of human brain neuronal pathways. Results and discussion One can merely suppose that its role in the BNST may be
potentially analogous to that revealed in the rodent brain. For instance, acute restrain stress is one of the main factors
activating nesfatinergic neurons in the rat hypothalamus
and selected brainstem structures, e.g., Edinger–Westphal,
locus coeruleus, and solitary tract nuclei (Stengel et al. 2010a; Goebel et al. 2009). Intracerebroventricular injec-
tion of nesfatin-1-elevated blood pressure (Yosten and
Samson 2009), whereas extended intraperitoneal admin-
istration of nesfatin-1 also facilitated anxiety in male rats
(Ge et al. 2015). Furthermore, animals exposed to acute,
but not chronic stress show an increase in both the mRNA
expression of NUCB2/nesfatin-1, and corticotropin-
releasing factor (CRF) within the hypothalamus (Xu et al. 2015). This may be especially important, given the distinct
population of stress-related CRF neurons, including some
oxytocin-regulated cells, is located within rat and human
lateral BNST (Janecek and Dabrowska 2018; Silberman
et al. 2013; Morin et al. 1999). Intriguingly, nesfatin-1/
NUCB2 mRNA expression in the human Edinger–West-
phal nuclei was significantly increased in suicidal victims
(with no diagnosed psychiatric disorders) among males,
whereas among females, this content was lower, com-
pared to controls (Bloem et al. 2012). The possible role
of nesfatin-1 signaling in the pathogenesis of sex-related Materials and methods Scale bars: 200 µm
(A, A1, B, B1, D), 100 µm (B2), 50 µm (B3) 1 3 353 The first identification of nesfatin-1-expressing neurons in the human bed nucleus of the stria… depressive-like and anxiety behavior in the context of
BNST function should be, therefore, taken into account. Given the small number of brain samples available for this
study, it is imperative that further studies expand these
findings to complement our initial report. obtain the density of immune-positive cells per standard-
ized area (0.2 mm2). Data are presented as a mean ± standard
error of the mean (SEM). References Endocrinology 151(7):3169–3180 Goebel M, Stengel A, Wang L, Lambrecht NW, Tache Y (2009) Nes-
fatin-1 immunoreactivity in rat brain and spinal cord autonomic
nuclei. Neurosci Lett 452(3):241–246 Rodríguez-Sierra OE, Goswami S, Turesson HK, Pare D (2016)
Altered responsiveness of BNST and amygdala neurons in trauma-
induced anxiety. Transl Psychiatry 6:e857 Goebel-Stengel M, Wang L (2013) Central and peripheral expres-
sion and distribution of NUCB2/nesfatin-1. Curr Pharm Des
19(39):6935–6940 Schalla MA, Stengel A (2018) Current understanding of the role of nes-
fatin-1. J Endocr Soc 2(10):1188–1206. https://doi.org/10.1210/
js.2018-00246f Gunay H, Tutuncu R, Aydin S, Dag E, Abasli D (2012) Decreased
plasma nesfatin-1 levels in patients with generalized anxiety dis-
order. Psychoneuroendocrinology 37(12):1949–1953 Schiltz K, Witzel J, Northoff G, Zierhut K, Gubka U, Fellman H, Kauf-
mann J, Tempelmann C, Wiebking C, Bogerts B (2007) Brain
pathology in pedophilic offenders: evidence of volume reduction
in the right amygdala and related diencephalic structures. Arch
Gen Psychiatry 64:737–746 Harris NA, Isaac AT, Günther A, Merkel K, Melchior J, Xu M,
Eguakun E, Perez R, Nabit BP, Flavin S, Gilsbach R, Shonesy B,
Hein L, Abel T, Baumann A, Matthews R, Centanni SW, Winder
DG (2018) Dorsal BNSTα2A-adrenergic receptors produce HCN-
dependent excitatory actions that initiate anxiogenic behaviors. J Neurosci 38(42):8922–8942. https://doi.org/10.1523/JNEUR
OSCI.0963-18.2018 Shimizu H, Mori M (2013) Nesfatin-1: its role in the diagnosis and
treatment of obesity and some psychiatric disorders. Methods Mol
Biol 963:327–338 Janeček M, Dabrowska J (2018) Oxytocin facilitates adaptive fear and
attenuates anxiety responses in animal models and human studies-
potential interaction with the corticotropin-releasing factor (CRF)
system in the bed nucleus of the stria terminalis (BNST). Cell
Tissue Res doi. https://doi.org/10.1007/s00441-018-2889-8 Silberman Y, Matthews RT, Winder DG (2013) A corticotrophin
releasing factor pathway for ethanol regulation of the ventral teg-
mental area in the bed nucleus of the stria terminalis. J Neurosci
33(3):950–960 Stengel A, Tache Y (2010) Nesfatin-1—role as possible new potent
regulator of food intake. Regul Pept 163:18–23 Jego S, Salvert D, Renouard L, Mori M, Goutagny R, Luppi PH, Fort
P (2012) Tuberal hypothalamic neurons secreting the satiety mol-
ecule nesfatin-1 are critically involved in paradoxical (REM) sleep
homeostasis. PLoS One 7:e52525 Stengel A, Tache Y (2013) Role of NUCB2/Nesfatin-1 in the hypo-
thalamic control of energy homeostasis. References Horm Metab Res
45:975–979 Kash TL, Pleil KE, Marcinkiewcz CA, Lowery-Gionta EG, Crowley
N, Mazzone C, Sugam J, Hardaway JA, McElligott ZA (2015)
Neuropeptide regulation of signaling and behavior in the BNST. Mol Cells 38(1):1–13 Stengel A, Goebel M, Wang L, Taché Y (2010a) Abdominal surgery
activates nesfatin-1 immunoreactive brain nuclei in rats. Peptides
31(2):263–270f Swaab D (2007) Sexual differentiation of the brain and behavior. Best
Pract Res Clin Endocrinol Metab 21(3):431–444 Mai J, Majtanik G, Paxinos C (2015) Atlas of the human brain, Fourth
ed., Academic Press, New York Theiss JD, Ridgewell C, McHugo M, Heckers S, Blackford JU (2017)
Manual segmentation of the human bed nucleus of the stria ter-
minalis using 3T MRI. Neuroimage 146:288–292 Martin LJ, Powers RE, Dellovade TL, Price DL (1991) The bed
nucleus-amygdala continuum in human and monkey. J Comp
Neurol 309(4):445–485 Vas S, Adori C, Konczol K, Katai Z, Pap D, Papp RS, Bagdy G, Palko-
vits M, Toth ZE (2013) Nesfatin-1/NUCB2 as a potential new
element of sleep regulation in rats. PLoS One 8:e59809f Merali Z, Cayer C, Kent P, Anisman H (2008) Nesfatin-1 increases
anxiety- and fear-related behaviors in the rat. Psychopharmacol-
ogy 201(1):115–123f Walter A, Mai JK, Lanta L, GörcsT (1991) Differential distribution
of immunohistochemical markers in the bednucleus of the stria
terminalis in the human brain. J Chem Neuroanat 4(4):281–289 Morin SM, Ling N, Liu XJ, Kahl SD, Gehlert DR (1999) Differential
distribution of urocortin- and corticotropin-releasing factor-like
immunoreactivities in the rat brain. Neuroscience 92(1):281–291 Weibert E, Hofmann T, Stengel A (2018) Role of nesfatin-1 in anxiety,
depression and the response to stress. Psychoneuroendocrinology
100:58–66. https://doi.org/10.1016/j.psyneuen.2018.09.037 Pałasz A, Krzystanek M, Worthington J, Czajkowska B, Kostro K,
Wiaderkiewicz R, Bajor G (2012) Nesfatin-1, a unique regulatory
neuropeptide of the brain. Neuropeptides 46(3):105–112 Wernecke K, Lamprecht I, Jöhren O, Lehnert H, Schulz C (2014) Nes-
fatin-1 increases energy expenditure and reduces food intake in
rats. Obesity (Silver Spring) 22(7):1662–1668 Pałasz A, Janas-Kozik M, Borrow A, Arias-Carrión O, Worthington JJ
(2018) The potential role of the novel hypothalamic neuropeptides
nesfatin-1, phoenixin, spexin and kisspeptin in the pathogenesis
of anxiety and anorexia nervosa. Neurochem Int 113:120–136 Xu YY, Ge JF, Qin G, Peng YN, Zhang CF, Liu XRLiang LC, Wang
ZZ, Chen FH, Li J (2015) Acute, but not chronic, stress increased
the plasma concentration and hypothalamic mRNA expression of
NUCB2/nesfatin-1 in rats. References Adrian TE, Allen JM, Bloom SR, Ghatei MA, Rossor MN, Roberts
GW, Crow TJ, Tatemoto K, Polak JM (1983) Neuropeptide Y
distribution in human brain. Nature 306(5943):584–586 Ari M, Ozturk OH, Bez Y, Oktar S, Erduran D (2011) High plasma
nesfatin-1 level in patients with major depressive disorder. Prog
Neuropsychopharmacol Biol Psychiatry 35(2):497–500 Avery SN, Clauss JA, Winder DG, Woodward N, Heckers S, Blackford
JU (2014) BNST neurocircuitry in humans. Neuroimage 91:311–
323. https://doi.org/10.1016/j.neuroimage.2014.01.017 Avery SN, Clauss JA, Blackford JU (2016) The human BNST: func-
tional role in anxiety and addiction. Neuropsychopharmacology
41(1):126–141. https://doi.org/10.1038/npp.2015.185 Bloem B, Xu L, Morava E, Faludi G, Palkovits M, Roubos EW, Kozicz
T (2012) Sex-specific differences in the dynamics of cocaine- and
amphetamine-regulated transcript and nesfatin-1 expressions in
the midbrain of depressed suicide victims vs. controls. Neurop-
harmacology 62(1):297–303 Ch’ng S, Fu J, Brown RM, McDougall SJ, Lawrence AJ (2018) The
intersection of stress and reward: BNST modulation of aversive
and appetitive states. Prog Neuropsychopharmacol Biol Psychiatry
87(Pt A):108–125. https://doi.org/10.1016/j.pnpbp.2018.01.005f Chung W, De Vries G, Swaab D (2002) Sexual differentiation of the
bed nucleus of the stria terminalis in humans may extend into
adulthood. J Neurosci 22(3):1027–1033 1 3 354 A. Pałasz et al. Pleil KE, Helms CM, Sobus JR, Daunais JB, Grant KA, Kash
TL (2016) Effects of chronic alcohol consumption on neu-
ronal function in the non-human primate BNST. Addict Biol
21(6):1151–1167 Dumais KM, Alonso AG, Immormino MA, Bredewold R, VeenemaAH
(2016) Involvement of the oxytocin system in the bed nucleus of
the stria terminalis in the sex-specific regulation of social recogni-
tion. Psychoneuroendocrinology 64:79–88 Emmerzaal TL, Kozicz T (2013) Nesfatin-1; implication in stress
and stress-associated anxiety and depression. Curr Pharm Des
19(39):6941–6948 Poulin JF, Arbour D, Laforest S, Drolet G (2009) Neuroanatomical
characterization of endogenous opioids in the bed nucleus of the
stria terminalis. Prog NeuropsychopharmacolBiol Psychiatry
33(8):1356–1365 Ge JF, Xu YY, Qin G, Pan XY, Cheng JQ, Chen FH (2015) Nesfatin-1,
a potent anorexic agent, decreases exploration and induces anx-
iety-like behavior in rats without altering learning or memory. Brain Res 1629:171–181 Ramanjaneya M, Chen J, Brown JE, Tripathi G, Hallschmid M, Patel S,
Kern W, Hillhouse EW, Lehnert H, Tan BK, Randeva HS (2010)
Identification of nesfatin-1 in human and murine adipose tissue:
a novel depot-specific adipokine with increased levels in obesity. Publisher’s Note Springer Nature remains neutral with regard to
jurisdictional claims in published maps and institutional affiliations. melanocortin system. Am J Physiol Regul Integr Comp Physiol
297(2):R330–R336 The first identification of nesfatin-1-expressing neurons in the human bed nucleus of the stria… References Neuropeptides 54:47–53 Palkovits M, Somogyvári-Vigh A, Arimura A (1995) Concentrations
of pituitary adenylate cyclase activating polypeptide (PACAP) in
human brain nuclei. Brain Res 699(1):116–120 Xu YY, Ge JF, Liang J, Cao Y, Shan F, Liu Y, Yan CY, Xia QR (2018)
Nesfatin-1 and cortisol: potential novel diagnostic biomarkers
in moderate and severe depressive disorder. Psychol Res Behav
Manag 11:495–502. https://doi.org/10.2147/PRBM.S183126 Pelrine E, Pasik SD, Bayat L, Goldschmiedt D, Bauer EP (2016)
5-HT2C receptors in the BNST are necessary for the enhance-
ment of fear learning by selective serotonin reuptake inhibitors. Neurobiol Learn Mem 136:189–195 Yosten GL, Samson WK (2009) Nesfatin-1 exerts cardiovascu-
lar actions in brain: possible interaction with the central 1 3 The first identification of nesfatin-1-expressing neurons in the human bed nucleus of the stria… 355 1 3
|
https://openalex.org/W3123009357
|
https://europepmc.org/articles/pmc7822156?pdf=render
|
English
| null |
C-Type Lectin CD209L/L-SIGN and CD209/DC-SIGN: Cell Adhesion Molecules Turned to Pathogen Recognition Receptors
| null | 2,020
|
cc-by
| 12,940
|
Citation: Rahimi, N. C-type Lectin
CD209L/L-SIGN
and CD209/DC-SIGN: Cell Adhesion
Molecules Turned to Pathogen
Recognition Receptors. Biology 2021,
10, 1. https://dx.doi.org/10.3390/
biology10010001 Received: 3 November 2020
Accepted: 16 December 2020
Published: 22 December 2020 Keywords: CD209; L-SIGN; CD209L; DC-SIGN; C-type lectin; cell adhesion molecule; C-type lectin
domain family 4 member M; CLEC4M; LSECtin; CLEC4G; SARS-CoV-2; COVID-19 Publisher’s Note: MDPI stays neu-
tral with regard to jurisdictional claims
in published maps and institutional
affiliations. Publisher’s Note: MDPI stays neu-
tral with regard to jurisdictional claims
in published maps and institutional
affiliations. biology biology biology biology Review
C-type Lectin CD209L/L-SIGN and CD209/DC-SIGN: Cell
Adhesion Molecules Turned to Pathogen Recognition Receptors Nader Rahimi Department of Pathology, School of Medicine, Boston University Medical Campus, Boston, MA 02118, USA;
nrahimi@bu.edu Simple Summary: COVID-19 pandemic continues to pose a serious threat to global public health
with overwhelming worldwide socio-economic disruption. SARS-CoV-2, the viral agent of COVID-
19, uses its surface glycoprotein Spike (S) for host cell attachment and entry. The emerging picture of
pathogenesis of SARS-CoV-2 demonstrates that S protein, in addition, to ACE2, interacts with the car-
bohydrate recognition domain (CRD) of C-type lectin receptors, CD209L and CD209. Recognition of
CD209L and CD209 which are widely expressed in SARS-CoV-2 target organs can facilitate entry
and transmission leading to dysregulation of the host immune response and other major organs
including, cardiovascular system. Establishing a comprehensive map of the SARS-CoV-2 interaction
with CD209 family proteins, and their roles in transmission and pathogenesis can provide new
insights into host-pathogen interaction with implications in therapies and vaccine development. Abstract: C-type lectin CD209/DC-SIGN and CD209L/L-SIGN proteins are distinct cell adhesion
and pathogen recognition receptors that mediate cellular interactions and recognize a wide range of
pathogens, including viruses such as SARS, SARS-CoV-2, bacteria, fungi and parasites. Pathogens
exploit CD209 family proteins to promote infection and evade the immune recognition system. CD209L and CD209 are widely expressed in SARS-CoV-2 target organs and can contribute to in-
fection and pathogenesis. CD209 family receptors are highly susceptible to alternative splicing
and genomic polymorphism, which may influence virus tropism and transmission in vivo. The car-
bohydrate recognition domain (CRD) and the neck/repeat region represent the key features of
CD209 family proteins that are also central to facilitating cellular ligand interactions and pathogen
recognition. While the neck/repeat region is involved in oligomeric dimerization, the CRD rec-
ognizes the mannose-containing structures present on specific glycoproteins such as those found
on the SARS-CoV-2 spike protein. Considering the role of CD209L and related proteins in diverse
pathogen recognition, this review article discusses the recent advances in the cellular and biochem-
ical characterization of CD209 and CD209L and their roles in viral uptake, which has important
implications in understanding the host–pathogen interaction, the viral pathobiology and driving
vaccine development of SARS-CoV-2. 2. CD209/DC-SIGN and CD209L/L-SIGN Family Proteins: Cell Adhesion Molecules
Turned to Pathogen Recognition Receptors: 2. CD209/DC-SIGN and CD209L/L-SIGN Family Proteins: Cell Adhesion Molecules
Turned to Pathogen Recognition Receptors: The conserved physiological function of CD209 family proteins is to mediate cell-cell
adhesion by functioning as high affinity receptors for intercellular adhesion molecules
2 and 3 (ICAM2 and ICAM3/ CD50) [17,18]. CD23 acts as a low-affinity receptor for
immunoglobulin E (IgE) and CR2/CD21 [19] and LSECtin interacts with CD44 on activated
T cells [20]. A survey of current literature indicates that these receptors are also among
the most common pathogen recognition receptors present in the human genome [21,22]. CD209L and CD209 serve as receptors for Ebolavirus [23], Hepatitis C virus [24], human
coronavirus 229E [25], human cytomegalovirus/HHV-5 [26], influenza virus [27], West-Nile
virus [26], Dengue virus [28] and Japanese encephalitis virus [29]. Recently, we and others
have shown that CD209 and CD209L is capable of recognizing SARS-CoV and SARS-
CoV-2 [16,26,30,31]. In addition to its ability to recognize a plethora of viruses, CD209
is also known to recognize parasites such as leishmania amastigotes [32] and Yersinia
pestis coccobacillus bacterium [33]. The complete list of viruses that are recognized by
CD209, CD209L and LSECtin are shown (Table 1). To date, it is not known whether CD23
is involved in any pathogen recognition. It is increasingly evident that viruses exploit host lectin receptors like the CD209L
family proteins and others for two major reasons; to promote infection of target cells
and evade the immune recognition system. In many cases lectin receptors such as CD209
and CD209L are employed as functional portals for viral recognition and infection. How-
ever, in some other cases, they may also enable infection of target cells via trans-infection
(i.e., cell captures the pathogen without entry and then passes it to another cell, which is
also a replication-independent mechanism [50]. For example, CD209 expressed in DCs
can bind to HIV envelope glycoprotein, gp120, without triggering cell-virus fusion [51]. The interaction of CD209 with gp120 appears to be complex as it can lead to both posi-
tive and negative outcomes for virus, perhaps depending to cell type in which CD209 is
expressed. In some cases, CD209-captured virions are internalized and targeted to the lyso-
some for degradation [52,53]. However, in cases which HIV-1 receptor and co-receptors
(CD4 and CCR5/CXCR4) are present on the host cells, CD209 can facilitate infection by
transferring the virus to immune cells [54,55]. 1. Introduction Lectins are a diverse family of carbohydrate recognizing proteins that possess car-
bohydrate -recognition domain (CRD) or sulfated glycosaminoglycan (SGAG)-binding
motif [1,2]. Derived from the Latin word “legere”, meaning “to select”. Lectins were
originally identified for their selective carbohydrate binding properties. However, now
it is known that they can also mediate protein-protein, protein-lipid or protein-nucleic
acid interactions [3]. By virtue of their CRD, lectins have the ability to recognize specific
carbohydrate structures on proteins, which in turn, mediate cell-cell and cell-pathogen
interactions [4,5]. There are currently fourteen structural families and three related subfam- Copyright: © 2020 by the author. Li-
censee MDPI, Basel, Switzerland. This
article is an open access article distributed
under the terms and conditions of the
Creative Commons Attribution (CC BY)
license (https://creativecommons.org/
licenses/by/4.0/). https://www.mdpi.com/journal/biology Biology 2021, 10, 1. https://dx.doi.org/10.3390/biology10010001 Biology 2021, 10, 1 2 of 15 ilies of lectins in human genome that span 76 different genes [6,7]. The C-type (calcium-
dependent) lectins with 66 gene members is one of the largest subgroups of the lectin
superfamily [6,8] that are further separated into multiple subgroups [7,9,10]. One of these
subgroups is the CD209/DC-SIGN (Dendritic cell-specific ICAM-3-grabbing non-integrin
1 also called CLEC4L, C-type lectin domain family 4 member L) subgroup, which includes
CD209/DC-SIGN (and three other member genes, namely CD209L/L-SIGN/CLEC4M
(Liver/lymph node-specific ICAM-3-grabbing non-integrin, C-type lectin domain family
4 member M), CD23 and LSECtin/CLEC4G [11,12]. Mouse genome encodes five homo-
logues of human CD209 with a variable sequence homology to human CD209 [13], but it
is not clear whether their function is similar to human CD209L and CD209. Other major
lectin subfamily proteins are the P-type lectins (mannose 6-phosphate (M6P) and the I-type
lectins. Siglecs (Sialic acid-binding immunoglobulin-type lectins) are the best characterized
I-type lectins [14,15]. Given the role of CD209L and related proteins in diverse mechanisms
of pathogen recognition and emerging evidence for the role of CD209 family proteins
in SARS-CoV-2 entry and infection [16], this review article particularly has focused on
the recent advances in the cellular and biochemical characterization of CD209 and CD209L
and their roles in virial uptake, which could provide valuable insights pertinent to current
pathobiological studies and therapeutic development of vaccines for SARS-Cov-2. 2. CD209/DC-SIGN and CD209L/L-SIGN Family Proteins: Cell Adhesion Molecules
Turned to Pathogen Recognition Receptors: Additionally, it was found that CD209-
dependent capture of HIV-1 virions could transiently protect virions from degradation,
which ultimately leads to viral infectivity [17,56–58], suggesting that CD209 positive DCs
capture and internalize HIV-1 virions and homes them to lymph nodes [59]. However,
the Trojan horse model of HIV transmission by CD209 was challenged by various stud-
ies [60]. Studies on B lymphocytes and platelets indicate that CD209 expressed in these Biology 2021, 10, 1 3 of 15 cells successfully mediate the entry of and infection by HIV-1 [61–63]. Similarly, CD209
and CD209L interact with Ebola virus glycoprotein and mediate infection of endothelial
cells via both cis- and trans-infection [23,36]. Likewise, the recent findings also support for
CD209L-mediated cis- and trans-infection of SARS-CoV-2 [16,64]. Aside from the role of
CD209 and CD209L in cis- and trans-infection and transmission, the recognition of these re-
ceptors by pathogens also can impact the host defense mechanism against these pathogens. For instance, CD209-dependent viral entry and infection can initiate signaling events in
host cells that compromise immune responses and promote infection of DCs [65,66]. In-
terestingly, although CD209L and CD209 are devoid of any enzymatic activity, however
upon interaction with pathogens, they can stimulate activation of multiple protein kinases,
GTPases and phosphatases [67]. Table 1. List of known pathogens recognized by CD209, CD209L and LSECtin lectin family proteins. The data is extracted from the publications available through PubMed. Table 1. List of known pathogens recognized by CD209, CD209L and LSECtin lectin family proteins. The data is extracted from the publications available through PubMed. The data is extracted from the publications available through PubMed. 2. CD209/DC-SIGN and CD209L/L-SIGN Family Proteins: Cell Adhesion Molecules
Turned to Pathogen Recognition Receptors: [49]
Gene Name
Pathogen Name
References
[49]
CD209
HIV-1 and HIV-2
[34,35]
[49]
Ebolavirus
[36,37]
[49]
Cytomegalovirus
[24,38]
[49]
Hepatitis C virus
[24]
[49]
Dengue virus
[28]
[49]
Measles virus
[39]
[49]
Herpes simplex virus
[40]
[49]
Influenza virus A
[27]
[49]
SARS-CoV-2
[16]
[49]
SARS-CoV
[30]
[49]
MERS
[41]
[49]
Japanese encephalitis virus
[29]
[49]
Lassa virus
[42]
[49]
Respiratory syncytial virus
[43]
[49]
Rift valley fever virus
[44]
[49]
Uukuniemi virus
[44]
[49]
West-Nile virus
[45]
[49]
CD209L
Ebolavirus
[23]
[49]
Hepatitis C virus
[24]
[49]
HIV-1
[37,46]
[49]
Human coronavirus 229E
[25]
[49]
Human
cytomegalovirus/HHV-5
[47]
[49]
Influenza virus
[27]
[49]
SARS-CoV
[26]
[49]
SARS-CoV-2
[16]
[49]
West-Nile virus
[26]
[49]
Japanese encephalitis virus
[29]
[49]
Marburg virus
[26]
[49]
LSECtin
Japanese encephalitis virus
[12]
[49]
Ebolavirus
[48]
[49]
SARS-CoV
[48]
[49]
Lassa virus
[49] CD209/DC-SIGN and CD209L/L-SIGN Family Proteins and Coronaviruses: 3. CD209/DC-SIGN and CD209L/L-SIGN Family Proteins and Coronaviruses: The evolutionarily conserved mechanism by which human coronaviruses including,
CoV-229E, NL63, OC43, HKU1, MERS-CoV, SARS-CoV, SARS-CoV-2 recognize the host
cells rely on the viral glycoprotein spike (S) that interacts with specific receptors on the host
target cells. Intriguingly, the S protein appears to be highly adept and can interact with
different types of host receptors. For examples, the S protein of CoV-229E and transmissible
gastroenteritis virus (TGEV) employ CD13 (aminopeptidase N) as a receptor for entry The evolutionarily conserved mechanism by which human coronaviruses including,
CoV-229E, NL63, OC43, HKU1, MERS-CoV, SARS-CoV, SARS-CoV-2 recognize the host
cells rely on the viral glycoprotein spike (S) that interacts with specific receptors on the host
target cells. Intriguingly, the S protein appears to be highly adept and can interact with
different types of host receptors. For examples, the S protein of CoV-229E and transmissible
gastroenteritis virus (TGEV) employ CD13 (aminopeptidase N) as a receptor for entry The evolutionarily conserved mechanism by which human coronaviruses including,
CoV-229E, NL63, OC43, HKU1, MERS-CoV, SARS-CoV, SARS-CoV-2 recognize the host
cells rely on the viral glycoprotein spike (S) that interacts with specific receptors on the host
target cells. Intriguingly, the S protein appears to be highly adept and can interact with
different types of host receptors. 2. CD209/DC-SIGN and CD209L/L-SIGN Family Proteins: Cell Adhesion Molecules
Turned to Pathogen Recognition Receptors: For examples, the S protein of CoV-229E and transmissible
gastroenteritis virus (TGEV) employ CD13 (aminopeptidase N) as a receptor for entry Biology 2021, 10, 1 4 of 15 and infection of target cells [68,69], whereas S protein of CoV-NL63 and HKU1 interact
with glycan-based receptors carrying 9-O-acetylated sialic acid (9-O-Ac-Sia) [70,71]. The S
protein of MERS-CoV uses Dipeptidyl peptidase 4 (DPP4/CD26) and carcinoembryonic
antigen-related cell adhesion molecule 5 (CEACAM5) as attachment or entry receptors
for infection [72,73]. CEACAM5 appears to facilitate MERS-CoV infection by enhancing
the attachment of the virus to the host cell surface [73]. The S protein of Filoviridae Marburg
virus, SARS-CoV [26,31] and SARS-CoV-2 [16,74] can employ the carbohydrate-recognition
domain (CRD) containing CD209L and CD209 lectins as attachment or entry receptor. (
)
g
y
p
CD209L is broadly expressed in human lung, kidney epithelium and endothelium [16]. Furthermore, human endothelial cells are permissive to SARS-CoV-2 infection and in-
terference with CD209L activity via shRNA or soluble CD209L inhibited SARS-CoV-2
entry and replication [16]. Remarkably, the S protein of human coronaviruses including,
NL63 [75], SARS-CoV [76] and SARS-CoV-2 [16] can also employ angiotensin-converting
enzyme 2(ACE2) as an entry receptor for infection, suggesting that both ACE2 and the lectin
family proteins, CD209L and CD209, contribute to the spread of these pathogens in vivo. Previous studies on SARS-CoV demonstrated a direct role for CD209L and CD209 in infec-
tion by acting as entry receptors for SARS independent of ACE2 [77–80]. Curiously, CD209L
can physically interact with ACE2 [16], suggesting both ACE2-dependent and independent
mechanisms for CD209L-mediated viral entry. However, the underlying mechanism of
CD209L and CD209 mediated SARS-CoV-2 infection is not fully understood and requires
further investigation. 4. Topology of CD209L/L-SIGN and CD209/D-SIGN CD209 family proteins are type II transmembrane glycoprotein receptors (i.e., C-
terminus is exposed outside the lipid bilayer and N-terminus resides in the cytosol). The ectodomain of CD209 and CD209L is composed of the neck region followed by the CRD. These domains represent the most distinct and functional features of these two receptors
(Figure 1A). The neck/repeat region is composed of 23 amino acids which is repeated
seven times in CD209L and CD209 and three times in CD23/FCER (Figure 1A). However,
the neck/repeat region on LSECtin/CLEC4G is replaced with a coil-coil motif, which is
also involved in protein-protein interaction (Figure 1A). Central to recognition of cellular
and pathogen glycoproteins, is the presence of the CRD on the C-terminus of CD209 family
proteins, which is paramount to recognition of mannose containing structures present on
specific glycoproteins. p
g y
p
CRD is a 110–130 amino acid long with a double-looped, two-stranded anti-parallel β-
sheet connected by two α-helices and a three-stranded anti-parallel β-sheet [81]. Typically,
CRD has two conserved disulfide bonds and up to four Ca2+ binding sites, depending on
the specific family of lectin. Amino acid residues with the carbonyl side chains are involved
in coordinating Ca2+ in the CRD, and these residues also directly bind to carbohydrates
leading to a ternary complex formation between a carbohydrate in a glycan, the Ca2+ ion,
and amino acids within the CRD. A typical CRD-carbohydrate interaction is shown (Figure
1B). Amino acid sequence alignment of CD209L with CD209 illustrates that these proteins
are highly conserved, suggesting that they likely evolved through gene duplications. There
are at least two putative internalization motifs at the cytosolic N-terminus tail of CD209
and CD209L, indicating that both CD209 and CD209L upon interaction with pathogens
are capable of undergoing internalization and delivering the pathogen inside the target
cells. The internalization motifs are di-leucine (LL) and tyrosine(Y)-based (Figure 2B),
but, the key tyrosine residue in the tyrosine-based internalization motif on the CD209L is
replaced with histidine (H) (Figure 2B), indicating that CD209L undergoes internalization
solely via di-leucine motif [82]. 5 of 15
ergoes
tion mo
9L unde Biology 2021, 10, 1 Figure 1. CD209 family proteins. (A) Graphic presentation of CD209 family proteins and the key domain information. The
schematic of domains do not directly correlate to the number of amino acids in each domain. 4. Topology of CD209L/L-SIGN and CD209/D-SIGN (B) Crystal structure of a
typical CRD complexed with carbohydrate and the position of the Ca2+ ion, which makes a tertiary complex between lectin
and carbohydrate structure. Figure 1. CD209 family proteins. (A) Graphic presentation of CD209 family proteins and the key domain information. The schematic of domains do not directly correlate to the number of amino acids in each domain. (B) Crystal structure of
a typical CRD complexed with carbohydrate and the position of the Ca2+ ion, which makes a tertiary complex between
lectin and carbohydrate structure. Figure 1. CD209 family proteins. (A) Graphic presentation of CD209 family proteins and the key domain information
schematic of domains do not directly correlate to the number of amino acids in each domain. (B) Crystal structure
typical CRD complexed with carbohydrate and the position of the Ca2+ ion, which makes a tertiary complex between l
and carbohydrate structure. Figure 1. CD209 family proteins. (A) Graphic presentation of CD209 family proteins and the key domain information. The
schematic of domains do not directly correlate to the number of amino acids in each domain. (B) Crystal structure of a
typical CRD complexed with carbohydrate and the position of the Ca2+ ion, which makes a tertiary complex between lectin
and carbohydrate structure. Figure 1. CD209 family proteins. (A) Graphic presentation of CD209 family proteins and the key domain information. The schematic of domains do not directly correlate to the number of amino acids in each domain. (B) Crystal structure of
a typical CRD complexed with carbohydrate and the position of the Ca2+ ion, which makes a tertiary complex between
lectin and carbohydrate structure. Figure 1. CD209 family proteins. (A) Graphic presentation of CD209 family proteins and the key domain informatio
schematic of domains do not directly correlate to the number of amino acids in each domain. (B) Crystal structu
typical CRD complexed with carbohydrate and the position of the Ca2+ ion, which makes a tertiary complex between
and carbohydrate structure. Figure 2. Amino acid sequence homology of CD209 and CD209L: (A) The schematic of CD209L is shown. 4. Topology of CD209L/L-SIGN and CD209/D-SIGN (B) Alignment
of the amino acids of human CD209 and CLEC4M (gene encoding for CD209L called C-type lectin domain family 4
A
SP|Q9NNX6|CD209_HUMAN MSDSKEPRLQQLGLLEEEQL--------RGLGFRQTRGYKSLAGCLGHGPLVLQLLSFTL 52
SP|Q9H2X3|CLC4M_HUMAN MSDSKEPRVQQLGLLEEDPTTSGIRLFPRDFQFQQIHGHKSSTGCLGHGALVLQLLSFML 60
SP|Q9NNX6|CD209_HUMAN LAGLL----VQVSKVPSSISQEQSRQDAIYQNLTQLKAAVGELSEKSKLQEIYQELTQLK 108
SP|Q9H2X3|CLC4M_HUMAN LAGVLVAILVQVSKVPSSLSQEQSEQDAIYQNLTQLKAAVGELSEKSKLQEIYQELTQLK 120
SP|Q9NNX6|CD209_HUMAN AAVGELPEKSKLQEIYQELTRLKAAVGELPEKSKLQEIYQELTWLKAAVGELPEKSKMQE 168
SP|Q9H2X3|CLC4M_HUMAN AAVGELPEKSKLQEIYQELTRLKAAVGELPEKSKLQEIYQELTRLKAAVGELPEKSKLQE 180
SP|Q9NNX6|CD209_HUMAN IYQELTRLKAAVGELPEKSKQQEIYQELTRLKAAVGELPEKSKQQEIYQELTRLKAAVGE 228
SP|Q9H2X3|CLC4M_HUMAN IYQELTRLKAAVGELPEKSKLQEIYQELTELKAAVGELPEKSKLQEIYQELTQLKAAVGE 240
SP|Q9NNX6|CD209_HUMAN LPEKSKQQEIYQELTQLKAAVERLCHPCPWEWTFFQGNCYFMSNSQRNWHDSITACKEVG 288
SP|Q9H2X3|CLC4M_HUMAN LPDQSKQQQIYQELTDLKTAFERLCRHCPKDWTFFQGNCYFMSNSQRNWHDSVTACQEVR 300
SP|Q9NNX6|CD209_HUMAN AQLVVIKSAEEQNFLQLQSSRSNRFTWMGLSDLNQEGTWQWVDGSPLLPSFKQYWNRGEP 348
SP|Q9H2X3|CLC4M_HUMAN AQLVVIKTAEEQNFLQLQTSRSNRFSWMGLSDLNQEGTWQWVDGSPLSPSFQRYWNSGEP 360
SP|Q9NNX6|CD209_HUMAN NNVGEEDCAEFSGNGWNDDKCNLAKFWICKKSAASCSRDEEQFLSPAPATPNPPPA 404
SP|Q9H2X3|CLC4M_HUMAN NNSGNEDCAEFSGSGWNDNRCDVDNYWICKKPAA-CFRDE---------------- 399
B
TM
Repeat region
Ectodomain
Cytoplasmic
domain
N
CRD
C
Endocytosis
signal
Endocytosis
signal
Calcium Binding
Disulfide Bond
N- Glycosylation
TM
Repeat Region
CRD
Figure 2. Amino acid sequence homology of CD209 and CD209L: (A) The schematic of CD209L is shown. (B) Alignm
of the amino acids of human CD209 and CLEC4M (gene encoding for CD209L called C-type lectin domain fam
A
SP|Q9NNX6|CD209_HUMAN MSDSKEPRLQQLGLLEEEQL--------RGLGFRQTRGYKSLAGCLGHGPLVLQLLSFTL 52
SP|Q9H2X3|CLC4M_HUMAN MSDSKEPRVQQLGLLEEDPTTSGIRLFPRDFQFQQIHGHKSSTGCLGHGALVLQLLSFML 60
SP|Q9NNX6|CD209_HUMAN LAGLL----VQVSKVPSSISQEQSRQDAIYQNLTQLKAAVGELSEKSKLQEIYQELTQLK 108
SP|Q9H2X3|CLC4M_HUMAN LAGVLVAILVQVSKVPSSLSQEQSEQDAIYQNLTQLKAAVGELSEKSKLQEIYQELTQLK 120
SP|Q9NNX6|CD209_HUMAN AAVGELPEKSKLQEIYQELTRLKAAVGELPEKSKLQEIYQELTWLKAAVGELPEKSKMQE 168
SP|Q9H2X3|CLC4M_HUMAN AAVGELPEKSKLQEIYQELTRLKAAVGELPEKSKLQEIYQELTRLKAAVGELPEKSKLQE 180
SP|Q9NNX6|CD209_HUMAN IYQELTRLKAAVGELPEKSKQQEIYQELTRLKAAVGELPEKSKQQEIYQELTRLKAAVGE 228
SP|Q9H2X3|CLC4M_HUMAN IYQELTRLKAAVGELPEKSKLQEIYQELTELKAAVGELPEKSKLQEIYQELTQLKAAVGE 240
SP|Q9NNX6|CD209_HUMAN LPEKSKQQEIYQELTQLKAAVERLCHPCPWEWTFFQGNCYFMSNSQRNWHDSITACKEVG 288
SP|Q9H2X3|CLC4M_HUMAN LPDQSKQQQIYQELTDLKTAFERLCRHCPKDWTFFQGNCYFMSNSQRNWHDSVTACQEVR 300
SP|Q9NNX6|CD209_HUMAN AQLVVIKSAEEQNFLQLQSSRSNRFTWMGLSDLNQEGTWQWVDGSPLLPSFKQYWNRGEP 348
SP|Q9H2X3|CLC4M_HUMAN AQLVVIKTAEEQNFLQLQTSRSNRFSWMGLSDLNQEGTWQWVDGSPLSPSFQRYWNSGEP 360
SP|Q9NNX6|CD209_HUMAN NNVGEEDCAEFSGNGWNDDKCNLAKFWICKKSAASCSRDEEQFLSPAPATPNPPPA 404
SP|Q9H2X3|CLC4M_HUMAN NNSGNEDCAEFSGSGWNDNRCDVDNYWICKKPAA-CFRDE---------------- 399
B
TM
Repeat region
Ectodomain
Cytoplasmic
domain
N
CRD
C
Endocytosis
signal
Endocytosis
signal
Calcium Binding
Disulfide Bond
N- Glycosylation
TM
Repeat Region
CRD
Figure 2. Amino acid sequence homology of CD209 and CD209L: (A) The schematic of CD209L is shown. (B) Alignment of
the amino acids of human CD209 and CLEC4M (gene encoding for CD209L called C-type lectin domain family 4 member M,
CLEC4M). The key common features of CD209L and CD209L, including potential PTMs and ion bindings are highlighted. lasmic
main
Cytoplasmic
domain A B QL--------R
sis Endocytosis
signal AGCLGHGPLVL
sis Endocytosis
signal Figure 2. Amino acid sequence homology of CD209 and CD209L: (A) The schematic of CD209L is shown. (B) Alignment
of the amino acids of human CD209 and CLEC4M (gene encoding for CD209L called C-type lectin domain family 4
Figure 2. Amino acid sequence homology of CD209 and CD209L: (A) The schematic of CD209L is shown. (B) Alignm
of the amino acids of human CD209 and CLEC4M (gene encoding for CD209L called C-type lectin domain fam
Figure 2. Amino acid sequence homology of CD209 and CD209L: (A) The schematic of CD209L is shown. (B) Alignment of
the amino acids of human CD209 and CLEC4M (gene encoding for CD209L called C-type lectin domain family 4 member M,
CLEC4M). The key common features of CD209L and CD209L, including potential PTMs and ion bindings are highlighted. 4. Topology of CD209L/L-SIGN and CD209/D-SIGN The ectodomain of CD209 and CD209L is composed of the neck region followed
by the CRD. These domains also represent the most distinct and functional features of
these two receptors. The neck region which is a repeat of 23 amino acids (Figure 2B),
is involved in protein dimerization/oligomerization [83,84], and may also contribute to
increased pathogen recognition and concentration of pathogens at the cell surface. The neck
region forms an α-helical coiled-coil fold that is thought to stabilize the oligomerization
of CD20 family proteins [85,86]. The presence of CRD on the ectodomain is paramount
to recognition of mannose, fucose- or galactose-containing structures on the pathogens Biology 2021, 10, 1 6 of 15 and the cellular ligands by CD209 and CD209L. It is thought that within the CRD a highly
conserved EPN motif (Glu-Pro-Asn) is responsible for recognition of mannose, fucose- or
galactose-containing structures [87]. Yet, despite a high degree of homology of the amino
acid residues in the CRD of CD209L and CD209, there is evidence for differential recognition
of oligosaccharide structures by these receptors. For example, CD209L appears to prefer
mannose oligosaccharides but not fucose-containing carbohydrates such as LewisX (LeX)
glycans [88]. Interestingly, a recent analysis revealed that N-glycosylation of SARS-CoV-2
spike protein is largely oligomannose-type glycans [89], which may account for the strong
binding of SARS-CoV-2 spike protein with CD209L and CD209 [16]. Furthermore, while
the ectodomain of CD209L contains two N-glycosylation sequons, at sites N92 and N361
(Figure 2B), only N92 is occupied. Curiously, removal of N-glycosylation on the CD209L increases the binding of CD209L
with the SARS-CoV-2 spike protein [16], suggesting that N-glycosylation of CD209L may
generate a hindrance for the CRD-mediated glycoprotein interaction [16] and may have
impact in virus tropism and transmissibility in vivo. A similar hindering mechanism for
ligand-receptor interaction by N-glycosylation was reported for an unrelated receptor
tyrosine kinase, vascular endothelial receptor-2 (VEGFR-2) interaction with its ligand [90]. Another important, yet poorly understood aspect of CD209 family proteins is their cyto-
plasmic N-terminus domain, which is vital for their signal transduction relays. To date,
there is no evidence for potential posttranslational modifications (PTMs) or a direct protein
interaction between the cytoplasmic N-terminus domains of CD209L and CD209 with
the signaling proteins [67]. 4. Topology of CD209L/L-SIGN and CD209/D-SIGN Unlike many of their counterpart receptors, the cytoplas-
mic N-terminus domains of CD209L and CD209 contain no conserved immunoreceptor
tyrosine-based inhibitory (ITIM, V/IXYXXL/I/V) motif, which interacts with the Src-
homology 2 (SH2) domain containing proteins [91]. While, the cytoplasmic N-terminus
domains of CD209L contains no tyrosine residue, the cytoplasmic N-terminus domain
of CD209 contains one tyrosine residue with a weak sequence homology to ITIM motif
(Figure 2B). But, there is no experimental evidence whether the key tyrosine (Y31) residue
is phosphorylated and recruits any SH2 domain signaling proteins to CD209. Furthermore,
there are multiple serine/threonine residues (four on the CD209 and 8 on the CD209L) on
the cytoplasmic N-terminus domains of CD209L and CD209 (Figure 2B), which potentially
could be phosphorylated. Similarly, there are multiple lysine (K) residues on the cytoplas-
mic N-terminus domain of CD209L and CD209 with potential to undergo ubiquitination. In particular, K5, which is conserved both in CD209L and CD209, has a high probability to
be ubiquitinated. Ubiquitin modification regulates both proteolytic and non-proteolytic
functions of proteins [92]. 5. Decoy CD209L and CD209 Proteins: These polymorphic changes most likely influence the physi-
ction of CD209L family proteins as well as the role that CD209L family pro-
binding to other pathogens, like SARS and SARS-CoV-2. Figure 3. The schematics of alternatively spliced variants of CD209L and CD209. Amino acid se-
quences of alternatively spliced variants of CD209L (A) and CD209 (B) were aligned via Clustal
Omega software program. The schematic of each alternatively variant proteins were presented. TM
Repeat
region
CRD
CD209L(Q9H2X3-6)
CD209L(Q9H2X3-7)
CD209L(Q9H2X3-8)
CD209L(Q9H2X3-9)
Ectodomain
Cytoplasmic
domain
CD209L(Q9H2X3-10)
CD209L(Q9H2X3-5)
CD209L(Q9H2X3-4)
CD209L(Q9H2X3-3)
CD209L(Q9H2X3-2)
CD209L(Q9H2X3)
TM
Repeat
region
CRD
CD209(Q9NNX6)
CD209 (Q9NNX6-2)
CD209 (Q9NNX6-3)
CD209 (Q9NNX6-4)
CD209 (Q9NNX6-6)
CD209 (Q9NNX6-7)
CD209 (Q9NNX6-8)
CD209 (Q9NNX6-9)
CD209 (Q9NNX6-10)
CD209 (Q9NNX6-11)
CD209 (Q9NNX6-12)
A
B
Figure 3. The schematics of alternatively spliced variants of CD209L and CD209. Amino acid sequences of alternatively
spliced variants of CD209L (A) and CD209 (B) were aligned via Clustal Omega software program. The schematic of each
alternatively variant proteins were presented. B B A Figure 3. The schematics of alternatively spliced variants of CD209L and CD209. Amino acid se-
quences of alternatively spliced variants of CD209L (A) and CD209 (B) were aligned via Clustal
Omega software program. The schematic of each alternatively variant proteins were presented. Figure 3. The schematics of alternatively spliced variants of CD209L and CD209. Amino acid sequences of alternatively
spliced variants of CD209L (A) and CD209 (B) were aligned via Clustal Omega software program. The schematic of each
alternatively variant proteins were presented. 5. Decoy CD209L and CD209 Proteins: A corollary to the function of CD209L and CD209 in pathogen and cellular ligand
recognition is that these receptors are highly susceptible to alternative splicing and ge-
nomic polymorphism, which may significantly influence their core functions. Analysis of
CD209L via uniprot (https://www.uniprot.org/), a freely accessible resource of protein
sequence, revealed that CD209L mRNA can generate at least 9 alternatively spliced variants
(Figure 3A). In many cases, the CRD is either completely or partially deleted (Figure 3A),
which generates a carbohydrate binding decoy of CD209L. In some other cases, the trans-
membrane domain is deleted, which results in the soluble form of CD209L. Yet in other
cases, multiple deletions occur simultaneously leading to generation of soluble CD209L
proteins without CRD (Figure 3A). Some of the alternatively spliced variants of CD209L
also carry a deletion on the neck region (Figure 3A). Similar to CD209L, CD209 also due
to alternative splice mechanism can also yield 11differant variants (Figure 3B). However,
the tissue expression profiles and the potential function of these alternatively spliced vari-
ants in normal cellular ligand recognition, and more importantly in pathogen interaction
remains largely unknown. Moreover, various recent studies have shown a distinct genomic
polymorphism, specifically, in the tandem-neck-repeat region of CD209L [93,94] and on Biology 2021, 10, 1 7 of 15 the promoter CD209 [95], which is linked to the pathogenesis of tuberculosis [96] and resis-
tance to HIV-1 [97]. These polymorphic changes most likely influence the physiological
function of CD209L family proteins as well as the role that CD209L family proteins play in
binding to other pathogens, like SARS and SARS-CoV-2. p
, p
y,
p
g
[
,
]
romoter CD209 [95], which is linked to the pathogenesis of tuberculosis [96]
ce to HIV-1 [97]. These polymorphic changes most likely influence the physi-
ction of CD209L family proteins as well as the role that CD209L family pro-
binding to other pathogens, like SARS and SARS-CoV-2. the promoter CD209 [95], which is linked to the pathogenesis of tuberculosis [96] and resis-
tance to HIV-1 [97]. These polymorphic changes most likely influence the physiological
function of CD209L family proteins as well as the role that CD209L family proteins play in
binding to other pathogens, like SARS and SARS-CoV-2. p
, p
y,
p
g
[
,
]
romoter CD209 [95], which is linked to the pathogenesis of tuberculosis [96]
ce to HIV-1 [97]. n Profile of CD209L Family Proteins in Human Tissues and Cells:
of the published data indicates that CD209/DC SIGN is predominantly ex
6. Expression Profile of CD209L Family Proteins in Human Tissues and Cells: In agreement with the recent
observations, our analysis of ACE2 mRNA through the Human Expression Atlas revealed Biology 2021, 10, 1 8 of 15 8 of 15 that ACE2 is highly expressed only in intestinal tissues (small intestine, colon and duo-
denum) and at the low levels in the hearth muscles, kidney, gallbladder and testis, but
undetectable in lung (Figure 4D). The observed limited/low ACE2 expression pattern in
human tissues suggests that SARS-CoV-2 could use alternative receptors for cell entry in
a cell-type dependent manner as CD209L and CD209 appear to be more broadly expressed
in human organs and tissues than the ACE2. Given the wider tissue expression patterns
of CD209 and CD209L in SARS-CoV-2 target tissues, they could play important roles in
the pathogenesis of SARS-CoV-2. he low levels in the hearth muscles, kidney, gallbladder and testis, but undetecta-
ng (Figure 4D). The observed limited/low ACE2 expression pattern in human tis-
gests that SARS-CoV-2 could use alternative receptors for cell entry in a cell-type
nt manner as CD209L and CD209 appear to be more broadly expressed in human
nd tissues than the ACE2. Given the wider tissue expression patterns of CD209
209L in SARS-CoV-2 target tissues, they could play important roles in the patho-
of SARS-CoV-2. that ACE2 is highly expressed only in intestinal tissues (small intestine, colon and duo-
denum) and at the low levels in the hearth muscles, kidney, gallbladder and testis, but
undetectable in lung (Figure 4D). The observed limited/low ACE2 expression pattern in
human tissues suggests that SARS-CoV-2 could use alternative receptors for cell entry in
a cell-type dependent manner as CD209L and CD209 appear to be more broadly expressed
in human organs and tissues than the ACE2. Given the wider tissue expression patterns
of CD209 and CD209L in SARS-CoV-2 target tissues, they could play important roles in
the pathogenesis of SARS-CoV-2. he low levels in the hearth muscles, kidney, gallbladder and testis, but undetecta
ng (Figure 4D). The observed limited/low ACE2 expression pattern in human tis-
gests that SARS-CoV-2 could use alternative receptors for cell entry in a cell-type
nt manner as CD209L and CD209 appear to be more broadly expressed in human
nd tissues than the ACE2. Given the wider tissue expression patterns of CD209
209L in SARS-CoV-2 target tissues, they could play important roles in the patho-
of SARS-CoV-2. Figure 4. Expression profile of CD209L family proteins and ACE2. g
g
CoV 2 CD290 Family Proteins and Endothelial Cells:
7. SARS-CoV-2, CD290 Family Proteins and Endothelial Cells: -CoV-2, CD290 Family Proteins and Endothelial Cells:
209L is expressed on endothelial cells of various organs including, the lymph
nuses [106], liver sinus endothelial cells [106], capillary endothelial cells of the
, the endothelial cells of the gastrointestinal tract [107], kidney endothelial cells
pulmonary endothelial cells [16,31,108]. In addition, other CD209L family lectin,
/CLEC4G is also expressed in liver sinusoidal endothelial cells and in the lymph
2]. CD209L expressed in endothelial cells can interact with ICAM3 which is ex-
in DCs, T-cells [109,110] and endothelial cells [18,111] generating multiple immu-
atory interactions between T-cells, DCs and endothelial cells (Figure 5). Moreover,
lial cells express both ICAM3 [18,111] and ICAM2 [112], capable of generating
pic interactions between endothelial cells (Figure 5) that regulate endothelial func-
angiogenesis. ICAM2 and ICAM3 belong to immunoglobulin (Ig) superfamily
CD209L is expressed on endothelial cells of various organs including, the lymph
nodes sinuses [106], liver sinus endothelial cells [106], capillary endothelial cells of the pla-
centa, the endothelial cells of the gastrointestinal tract [107], kidney endothelial cells [16]
and pulmonary endothelial cells [16,31,108]. In addition, other CD209L family lectin,
LSECtin/CLEC4G is also expressed in liver sinusoidal endothelial cells and in the lymph
node [12]. CD209L expressed in endothelial cells can interact with ICAM3 which is ex-
pressed in DCs, T-cells [109,110] and endothelial cells [18,111] generating multiple im-
munoregulatory interactions between T-cells, DCs and endothelial cells (Figure 5). More-
over, endothelial cells express both ICAM3 [18,111] and ICAM2 [112], capable of generating
homotypic interactions between endothelial cells (Figure 5) that regulate endothelial func-
tion and angiogenesis. ICAM2 and ICAM3 belong to immunoglobulin (Ig) superfamily
cell adhesion molecules. ICAM2 and ICAM3 are highly glycosylated at the ectodomain
and contain two and five Ig domains, respectively. Consistent with this idea, loss of
ICAM2 in mouse results in impaired endothelial cell migration and angiogenesis [112]. However, expression of ICAM2, ICAM3 and CD209L in endothelial cells, the role of this
CD209L/ICAM2/ICAM3 axis in endothelial biology is largely unknown. A recent study
demonstrated that knocked down of CD209L by shRNA in immortalized endothelial
cells increased cell migration, but inhibited capillary tube formation/in vitro angiogen-
esis [16], underscoring the importance of CD209L pathway in the regulation of normal
endothelial function. Recent studies demonstrate that human vascular system is a direct target of SARS-
CoV2 [16,113,114]. n Profile of CD209L Family Proteins in Human Tissues and Cells:
of the published data indicates that CD209/DC SIGN is predominantly ex
6. Expression Profile of CD209L Family Proteins in Human Tissues and Cells: (A-D) Data were extracted from
the Human Expression Atlas (data was accessed in 11/03/2020) [101]. Arrowhead points to expres-
Figure 4. Expression profile of CD209L family proteins and ACE2. (A-D) Data were extracted from the Human Expression
Atlas (data was accessed in 11/03/2020) [101]. Arrowhead points to expression of individual genes in lung. Figure 4. Expression profile of CD209L family proteins and ACE2. (A-D) Data were extracted from
the Human Expression Atlas (data was accessed in 11/03/2020) [101]. Arrowhead points to expres-
Figure 4. Expression profile of CD209L family proteins and ACE2. (A-D) Data were extracted from the Human Expression
Atlas (data was accessed in 11/03/2020) [101]. Arrowhead points to expression of individual genes in lung. n Profile of CD209L Family Proteins in Human Tissues and Cells:
of the published data indicates that CD209/DC SIGN is predominantly ex
6. Expression Profile of CD209L Family Proteins in Human Tissues and Cells: of the published data indicates that CD209/DC-SIGN is predominantly ex-
he monocyte-derived dendritic cells (mo-DCs), and on mo-DCs of immature
n lymphoid tissue, lymph nodes and spleen [18,98], whereas CD209L is pre-
expressed in human type II alveolar cells of lung, liver, kidney and lymph
,31,99]. Our analysis of expression of CD209L, CD209 and LSECtin mRNAs
Human Expression Atlas, a publically available dataset [100], revealed that
ression is relatively restricted to a few human organs. The highest levels of
s observed in liver and lymph node followed by placenta, lung and ovary
CD209, on the other hand, is broadly expressed in human tissues and organs
s levels. The highest levels of CD209 expression was observed in the lymph
Survey of the published data indicates that CD209/DC-SIGN is predominantly ex-
pressed on the monocyte-derived dendritic cells (mo-DCs), and on mo-DCs of immature
and mature in lymphoid tissue, lymph nodes and spleen [18,98], whereas CD209L is pre-
dominantly expressed in human type II alveolar cells of lung, liver, kidney and lymph
nodes [16,18,31,99]. Our analysis of expression of CD209L, CD209 and LSECtin mRNAs
through the Human Expression Atlas, a publically available dataset [100], revealed that
CD209L expression is relatively restricted to a few human organs. The highest levels of
CD209L was observed in liver and lymph node followed by placenta, lung and ovary
(Figure 4A). CD209, on the other hand, is broadly expressed in human tissues and organs
at the various levels. The highest levels of CD209 expression was observed in the lymph
node followed by the adipose tissue, small intestine and rectum (Figure 4B). Similar
to CD209, LSECtin/CLEC4G was also broadly expressed in human tissues and organs. The highest levels of LSECtin was observed in liver and lymph node, followed with DCs,
monocytes, adipose tissue, heart muscle and cerebellum (Figure 4C). Considering the cur-
rent SARS-CoV-2 pandemic, it is worthwhile to compare the expression profiles CD209
family proteins with ACE2. Human ACE2 is considered an important entry receptor for
SARS-CoV-2 [76,101,102] and was previously reported to be widely expressed in the lung,
vascular system and other organs [103]. However, a recent study demonstrated that ACE2
is expressed at very low levels and only in a small subset of lung epithelial cells [104]
and low-to-undetectable levels in endothelial cells [105]. g
g
CoV 2 CD290 Family Proteins and Endothelial Cells:
7. SARS-CoV-2, CD290 Family Proteins and Endothelial Cells: Although primarily infects the lungs, the SARS-CoV-2 virus also tar-
gets multiple other organs including, the cardiovascular system, gastrointestinal tract,
and the kidneys [115–118]. Severe endothelial cell injury, vascular thrombosis with micro-
angiopathy, occlusion of alveolar capillaries, and angiogenesis are commonly observed Biology 2021, 10, 1
Biology 20 9 of 15 9 of 15 in COVID-19 patients [114]. COVID-19 patients suffer from distinct endothelial cell in-
jury (i.e., endothelitis), and altered angiogenesis with widespread microvascular thrombo-
sis [119,120]. These observations coupled with the fact that vascular endothelial dysfunction
also plays crucial roles in the pathogenesis of COVID-19 [22,121], underscores the piv-
otal role of endothelial cells in the pathobiology of SARS-CoV-2 infection. Collectively,
the data points to potentially important role for CD209 family proteins in the pathogenesis
of SARS-CoV-2. Therefore, CD209L not only can act as a SARS-COV-2 entry receptor, but
also performs critical functions in the angiogenic responses of endothelial cells, suggesting
that SARS-COV-2, by exploiting CD209L, could undermine CD209L function in endothelial
cells, leading to endothelial cell injury and altered angiogenesis. cell adhesion molecules. ICAM2 and ICAM3 are highly glycosylated at the ectod
and contain two and five Ig domains, respectively. Consistent with this idea, l
ICAM2 in mouse results in impaired endothelial cell migration and angiogenesis
However, expression of ICAM2, ICAM3 and CD209L in endothelial cells, the role
CD209L/ICAM2/ICAM3 axis in endothelial biology is largely unknown. A recent
demonstrated that knocked down of CD209L by shRNA in immortalized endothelia
increased cell migration, but inhibited capillary tube formation/in vitro angiogenes
underscoring the importance of CD209L pathway in the regulation of normal endo
function. in COVID-19 patients [114]. COVID-19 patients suffer from distinct endothelial cell in-
jury (i.e., endothelitis), and altered angiogenesis with widespread microvascular thrombo-
sis [119,120]. These observations coupled with the fact that vascular endothelial dysfunction
also plays crucial roles in the pathogenesis of COVID-19 [22,121], underscores the piv-
otal role of endothelial cells in the pathobiology of SARS-CoV-2 infection. Collectively,
the data points to potentially important role for CD209 family proteins in the pathogenesis
of SARS-CoV-2. Therefore, CD209L not only can act as a SARS-COV-2 entry receptor, but
also performs critical functions in the angiogenic responses of endothelial cells, suggesting
that SARS-COV-2, by exploiting CD209L, could undermine CD209L function in endothelial
cells, leading to endothelial cell injury and altered angiogenesis. cell adhesion molecules. g
g
CoV 2 CD290 Family Proteins and Endothelial Cells:
7. SARS-CoV-2, CD290 Family Proteins and Endothelial Cells: ICAM2 and ICAM3 are highly glycosylated at the ectodo
and contain two and five Ig domains, respectively. Consistent with this idea, l
ICAM2 in mouse results in impaired endothelial cell migration and angiogenesis
However, expression of ICAM2, ICAM3 and CD209L in endothelial cells, the role
CD209L/ICAM2/ICAM3 axis in endothelial biology is largely unknown. A recent
demonstrated that knocked down of CD209L by shRNA in immortalized endothelia
increased cell migration, but inhibited capillary tube formation/in vitro angiogenes
underscoring the importance of CD209L pathway in the regulation of normal endo
function. Figure 5. CD209L/L-SIGN mediates multiple cell-cell interactions between endothelial c
and T-cells. CD209L cellular ligands (ICAM2 and ICAM3) are expressed in DCs, T-cells
thelial cells leading multiple immunoregulatory and other interactions between T-cells,
Figure 5. CD209L/L-SIGN mediates multiple cell-cell interactions between endothelial cells, DCs and T-cells. CD209L
cellular ligands (ICAM2 and ICAM3) are expressed in DCs, T-cells and endothelial cells leading multiple immunoregulatory
and other interactions between T-cells, DCs and endothelial cells. CD209L/ICAM2/ICAM3 interactions in endothelial cells
can regulate endothelial adhesion and angiogenic properties of endothelial cells. Figure 5. CD209L/L-SIGN mediates multiple cell-cell interactions between endothelial
and T-cells. CD209L cellular ligands (ICAM2 and ICAM3) are expressed in DCs, T-cells
thelial cells leading multiple immunoregulatory and other interactions between T-cells
Figure 5. CD209L/L-SIGN mediates multiple cell-cell interactions between endothelial cells, DCs and T-cells. CD209L
cellular ligands (ICAM2 and ICAM3) are expressed in DCs, T-cells and endothelial cells leading multiple immunoregulatory
and other interactions between T-cells, DCs and endothelial cells. CD209L/ICAM2/ICAM3 interactions in endothelial cells
can regulate endothelial adhesion and angiogenic properties of endothelial cells. 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] Park, C.G.; Takahara, K.; Umemoto, E.; Yashima, Y.; Matsubara, K.; Matsuda, Y.; Clausen, B.E.; Inaba
mouse homologues of the human dendritic cell C-type lectin, DC-SIGN. Int. Immunol. 2001, 13, 1283–12 13. Park, C.G.; Takahara, K.; Umemoto, E.; Yashima, Y.; Matsubara, K.; Matsuda, Y.; Clausen, B.E.; Inaba, K.; Steinman, R.M. Five
mouse homologues of the human dendritic cell C-type lectin, DC-SIGN. Int. Immunol. 2001, 13, 1283–1290. [CrossRef] [PubMed]
14. Crocker, P.R.; Clark, E.A.; Filbin, M.; Gordon, S.; Jones, Y.; Kehrl, J.H.; Kelm, S.; Le Douarin, N.; Powell, L.; Roder, J.; et al. Siglecs:
A family of sialic-acid binding lectins. Glycobiology 1998, 8. [CrossRef] o se
o
o og es o
e
e
c ce
C ype ec
,
C S GN
00 ,
,
83
90 [C oss e ] [
Me ]
14. Crocker, P.R.; Clark, E.A.; Filbin, M.; Gordon, S.; Jones, Y.; Kehrl, J.H.; Kelm, S.; Le Douarin, N.; Powell, L.; Roder, J.; et al. Siglecs:
A family of sialic-acid binding lectins. Glycobiology 1998, 8. [CrossRef] g
yp
14. Crocker, P.R.; Clark, E.A.; Filbin, M.; Gordon, S.; Jones, Y.; Kehrl, J.H.; Kelm, S.; Le Douarin, N.; Pow
A family of sialic-acid binding lectins. Glycobiology 1998, 8. [CrossRef] y
g
y
gy
15. Varki, A.; Angata, T. Siglecs—The major subfamily of I-type lectins. Glycobiology 2006, 16, 1R–27R. [CrossRef] [PubMed]
16. Amraie, R.; Napoleon, M.A.; Yin, W.; Berrigan, J.; Suder, E.; Zhao, G.; Olejnik, J.; Gummuluru, S.; Muhlberger, E.; Chitalia, V.; et al. CD209L/L-SIGN and CD209/DC-SIGN act as receptors for SARS-CoV-2 and are differentially expressed in lung and kidney y
g
15. Varki, A.; Angata, T. Siglecs—The major subfamily of I-type lectins. Glycobiology 2006, 16, 1R–27R. [CrossRef] [PubMed]
16. Amraie, R.; Napoleon, M.A.; Yin, W.; Berrigan, J.; Suder, E.; Zhao, G.; Olejnik, J.; Gummuluru, S.; Muhlberger, E.; Chitalia, V.; et al. 15. Varki, A.; Angata, T. Siglecs—The major subfamily of I-type lectins. Glycob 15. Varki, A.; Angata, T. Siglecs—The major subfamily of I-type lectins. Glycobiology 2006, 16, 1R–27R. [CrossRef] [PubMed] 16. Amraie, R.; Napoleon, M.A.; Yin, W.; Berrigan, J.; Suder, E.; Zhao, G.; Olejnik, J.; Gummuluru, S.; Muhlberger, E.; Chitalia, V.; et al. CD209L/L-SIGN and CD209/DC-SIGN act as receptors for SARS-CoV-2 and are differentially expressed in lung and kidney
epithelial and endothelial cells. bioRxiv 2020. [CrossRef] p
17. Geijtenbeek, T.B.; Torensma, R.; van Vliet, S.J.; van Duijnhoven, G.C.; Adema, G.J.; van Kooyk, Y.; Figdor, C.G. adhesion and angiogenic pro
8. Conclusions and Perspective: Funding: This work was supported in part through grants from CTSI grant UL1 TR001430, Mal-
ory Fund, Department of Pathology, Boston University and a grant from The Evans Center for
Interdisciplinary Biomedical Research ARC on COVID-19. Acknowledgments: Author thanks Razie Amraei and Rachel Ho for reading and commenting on
the manuscript. Conflicts of Interest: The author declares no conflict of interest. Conflicts of Interest: The author declares no conflict of interest. adhesion and angiogenic pro
8. Conclusions and Perspective: Recent studies demonstrate that human vascular system is a direct target of
CoV2 [16,113,114]. Although primarily infects the lungs, the SARS-CoV-2 virus al
gets multiple other organs including, the cardiovascular system, gastrointestinal tra
the kidneys [115–118]. Severe endothelial cell injury, vascular thrombosis with mic
giopathy, occlusion of alveolar capillaries, and angiogenesis are commonly obser
COVID-19 patients [114]. COVID-19 patients suffer from distinct endothelial cell
(i.e., endothelitis), and altered angiogenesis with widespread microvascular throm
[119,120]. These observations coupled with the fact that vascular endothelial dysfu
also plays crucial roles in the pathogenesis of COVID-19 [22,121], underscores the p
role of endothelial cells in the pathobiology of SARS-CoV-2 infection. Collective
data points to potentially important role for CD209 family proteins in the pathogen
SARS-CoV-2. Therefore, CD209L not only can act as a SARS-COV-2 entry recepto
also performs critical functions in the angiogenic responses of endothelial cells, sugg
that SARS COV 2 by exploiting CD209L could undermine CD209L function in en
Infectious diseases have been foremost among the threats posed to human health
and survival through history. The novel coronavirus disease-2019 (COVID-19) pandemic
continues to pose a serious threat to global public health with overwhelming worldwide
socio-economic disruption. The emerging picture of pathogenesis of SARS-CoV-2 is that
in addition to dysregulation of the host immune response in lung, also targets other
major organs including, cardiovascular system and others, which may account for COVID-
19 induced mortality. The vascular system, particularly, the pulmonary endothelium
may play a pivotal role in the pathogenesis of COVID-19 via engagement of CD209L
and other related receptors. To date, many aspects of SARS-CoV-2 transmission, infection,
and treatment remain unclear. Not only can CD209L function as an entry receptor it can
also contribute to the pathogenesis of COVID-19. Establishing a comprehensive map of
the SARS-CoV-2 interaction with CD209 family proteins, and their roles in endothelial
functions and injury can provide new insights into pathogenesis of COVID-19 and offers
a bona fide treatment modality. Biology 2021, 10, 1 10 of 15 10 of 15 Author Contributions: N.R. conceived and wrote the manuscript. The author has read and agreed
to the published version of the manuscript. Funding: This work was supported in part through grants from CTSI grant UL1 TR001430, Mal-
ory Fund, Department of Pathology, Boston University and a grant from The Evans Center for
Interdisciplinary Biomedical Research ARC on COVID-19. 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] 1. Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] 2. Esko, J.D.; Selleck, S.B. Order out of chaos: Assembly of ligand binding sites in heparan sulfate. Ann. Rev. Biochem. 2002, 71,
435–471. [CrossRef] [PubMed] 3. Kilpatrick, D.C. Animal lectins: A historical introduction and overview. Biochim. Biophys. Acta 2002, 1572, 187–197. [CrossRef]
4. Van Breedam, W.; Pohlmann, S.; Favoreel, H.W.; de Groot, R.J.; Nauwynck, H.J. Bitter-sweet symphony: Glycan-lectin interactions
in virus biology. FEMS Microbiol. Rev. 2014, 38, 598–632. [CrossRef] 5. Sharon, N.; Lis, H. History of lectins: From hemagglutinins to biological recognition molecules. Glycobiology 2004, 14, 53R–62R. [CrossRef] 6. Drickamer, K.; Fadden, A.J. Genomic analysis of C-type lectins. Biochem. Soc. Symp. 2002, 69, 59–72 7. Zelensky, A.N.; Gready, J.E. The C-type lectin-like domain superfamily. FEBS J. 2005, 272, 6179–621 7. Zelensky, A.N.; Gready, J.E. The C-type lectin-like domain superfamily. FEBS J. 2005, 272, 6179–6217. [CrossRef]
h
h
l b
f
b h d
b
l
d
d
l
[ 8. Sharon, N.; Lis, H. The structural basis for carbohydrate recognition by lectins. Adv. Exp. Med. Biol. 20 Lis, H. The structural basis for carbohydrate recognition by lectins. Adv. Exp. Med. Biol. 2001, 491, 1–16. [Cro Lis, H. The structural basis for carbohydrate recogniti ,
;
,
y
g
y
p
,
,
[
]
9. Ebner, S.; Sharon, N.; Ben-Tal, N. Evolutionary analysis reveals collective properties and specificity in the C-type lectin and lectin-
like domain superfamily. Proteins 2003, 53, 44–55. [CrossRef] 10. Drickamer, K.; Taylor, M.E. Recent insights into structures and functions of C-type lectins in the immune system. Curr. Opin. Struct. Biol. 2015, 34, 26–34. [CrossRef] 11. Soilleux, E.J.; Barten, R.; Trowsdale, J. DC-SIGN; a related gene, DC-SIGNR and CD23 form a cluster on
165, 2937–2942. [CrossRef] [PubMed] , R.; Trowsdale, J. DC-SIGN; a related gene, DC-SIGNR and CD23 form a cluster on 19p13. J. Immunol. 2000
ossRef] [PubMed] 12. Liu, W.; Tang, L.; Zhang, G.; Wei, H.; Cui, Y.; Guo, L.; Gou, Z.; Chen, X.; Jiang, D.; Zhu, Y.; et al. Characterization of a novel
C-type lectin-like gene, LSECtin: Demonstration of carbohydrate binding and expression in sinusoidal endothelial cells of liver
and lymph node. J. Biol. Chem. 2004, 279, 18748–18758. [CrossRef] [PubMed] 13. 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] Identification
of DC-SIGN, a novel dendritic cell-specific ICAM-3 receptor that supports primary immune responses. Cell 2000, 100, 575–585. [CrossRef] 18. Bashirova, A.A.; Geijtenbeek, T.B.; van Duijnhoven, G.C.; van Vliet, S.J.; Eilering, J.B.; Martin, M.P.; Wu, L.; Martin, T.D.; Viebig,
N.; Knolle, P.A.; et al. A dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN)-related protein
is highly expressed on human liver sinusoidal endothelial cells and promotes HIV-1 infection. J. Exp. Med. 2001, 193, 671–678. [CrossRef] 19. Hibbert, R.G.; Teriete, P.; Grundy, G.J.; Beavil, R.L.; Reljic, R.; Holers, V.M.; Hannan, J.P.; Sutton, B.J.; Gould, H.J.; McDonnell, J.M. The structure of human CD23 and its interactions with IgE and CD21. J. Exp. Med. 2005, 202, 751–760. [CrossRef] g
20. Tang, L.; Yang, J.; Tang, X.; Ying, W.; Qian, X.; He, F. The DC-SIGN family member LSECtin is a nove
T cells. Eur. J. Immunol. 2010, 40, 1185–1191. [CrossRef] 21. Kerr, J.R. Cell adhesion molecules in the pathogenesis of and host defence against microbial infection. Mol. Pathol. 1999, 52,
220–230. [CrossRef] [PubMed] 22. Amraei, R.; Rahimi, N. COVID-19, Renin-Angiotensin System and Endothelial Dysfunction. Cells 2020, 9, 1652. [CrossRef]
[PubMed] 23. Alvarez, C.P.; Lasala, F.; Carrillo, J.; Muniz, O.; Corbi, A.L.; Delgado, R. C-type lectins DC-SIGN and L-SIGN mediate cellular
entry by Ebola virus in cis and in trans. J. Virol. 2002, 76, 6841–6844. [CrossRef] [PubMed] 11 of 15 11 of 15 Biology 2021, 10, 1 24. Cormier, E.G.; Durso, R.J.; Tsamis, F.; Boussemart, L.; Manix, C.; Olson, W.C.; Gardner, J.P.; Dragic, T. L-SIGN (CD209L) and DC-
SIGN (CD209) mediate transinfection of liver cells by hepatitis C virus. Proc. Natl. Acad. Sci. USA 2004, 101, 14067–14072. [CrossRef] [PubMed] [
] [
]
25. Jeffers, S.A.; Hemmila, E.M.; Holmes, K.V. Human coronavirus 229E can use CD209L (L-SIGN) to enter cells. Adv. Exp. Med. Biol. 2006, 581, 265–269. [CrossRef] 26. Marzi, A.; Gramberg, T.; Simmons, G.; Moller, P.; Rennekamp, A.J.; Krumbiegel, M.; Geier, M.; Eisemann, J.; Turza, N.; Saunier,
B.; et al. DC-SIGN and DC-SIGNR interact with the glycoprotein of Marburg virus and the S protein of severe acute respiratory
syndrome coronavirus. J. Virol. 2004, 78, 12090–12095. [CrossRef] y
27. Londrigan, S.L.; Turville, S.G.; Tate, M.D.; Deng, Y.M.; Brooks, A.G.; Reading, P.C. N-linked glycosylation facilitates sialic
acid-independent attachment and entry of influenza A viruses into cells expressing DC-SIGN or L-SIGN. J. Virol. 2011, 85,
2990–3000. [CrossRef] 28. 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] Tassaneetrithep, B.; Burgess, T.H.; Granelli-Piperno, A.; Trumpfheller, C.; Finke, J.; Sun, W.; Eller, M.A.; Pattanapanyasat, K.;
Sarasombath, S.; Birx, D.L.; et al. DC-SIGN (CD209) mediates dengue virus infection of human dendritic cells. J. Exp. Med. 2003,
197, 823–829. [CrossRef] 29. Shimojima, M.; Takenouchi, A.; Shimoda, H.; Kimura, N.; Maeda, K. Distinct usage of three C-type lectins by Japanese encephalitis
virus: DC-SIGN, DC-SIGNR, and LSECtin. Arch. Virol. 2014, 159, 2023–2031. [CrossRef] 30. Yang, Z.Y.; Huang, Y.; Ganesh, L.; Leung, K.; Kong, W.P.; Schwartz, O.; Subbarao, K.; Nabel, G.J. pH-dependent entry of severe
acute respiratory syndrome coronavirus is mediated by the spike glycoprotein and enhanced by dendritic cell transfer through
DC-SIGN. J. Virol. 2004, 78, 5642–5650. [CrossRef] J
31. Jeffers, S.A.; Tusell, S.M.; Gillim-Ross, L.; Hemmila, E.M.; Achenbach, J.E.; Babcock, G.J.; Thomas, W.D., Jr.; Thackray, L.B.; Young,
M.D.; Mason, R.J.; et al. CD209L (L-SIGN) is a receptor for severe acute respiratory syndrome coronavirus. Proc. Natl. Acad. Sci. USA 2004, 101, 15748–15753. [CrossRef] [PubMed] 32. Colmenares, M.; Puig-Kroger, A.; Pello, O.M.; Corbi, A.L.; Rivas, L. Dendritic cell (DC)-specific intercellular adhesion molecule
3 (ICAM-3)-grabbing nonintegrin (DC-SIGN, CD209), a C-type surface lectin in human DCs, is a receptor for Leishmania
amastigotes. J. Biol. Chem. 2002, 277, 36766–36769. [CrossRef] [PubMed] 33. Zhang, P.; Skurnik, M.; Zhang, S.S.; Schwartz, O.; Kalyanasundaram, R.; Bulgheresi, S.; He, J.J.; Klena, J.D.; Hinnebusch, B.J.;
Chen, T. Human dendritic cell-specific intercellular adhesion molecule-grabbing nonintegrin (CD209) is a receptor for Yersinia
pestis that promotes phagocytosis by dendritic cells. Infect. Immun. 2008, 76, 2070–2079. [CrossRef] [PubMed] g
y
y
34. Curtis, B.M.; Scharnowske, S.; Watson, A.J. Sequence and expression of a membrane-associated C-type lectin that exhibits
CD4-independent binding of human immunodeficiency virus envelope glycoprotein gp120. Proc. Natl. Acad. Sci. USA 1992, 89,
8356–8360. [CrossRef] [
]
35. Geijtenbeek, T.B.H.; van Duijnhoven, G.C.F.; van Vliet, S.J.; Krieger, E.; Vriend, G.; Figdor, C.G.; van Kooyk, Y. Identification of
different binding sites in the dendritic cell-specific receptor DC-SIGN for intercellular adhesion molecule 3 and HIV-1. J. Biol. Chem. 2002, 277, 11314–11320. [CrossRef] [
]
36. Simmons, G.; Reeves, J.D.; Grogan, C.C.; Vandenberghe, L.H.; Baribaud, F.; Whitbeck, J.C.; Burke, E.; Buchmeier, M.J.; Soilleux,
E.J.; Riley, J.L.; et al. DC-SIGN and DC-SIGNR bind ebola glycoproteins and enhance infection of macrophages and endothelial
cells. Virology 2003, 305, 115–123. [CrossRef] gy
,
,
[
]
37. 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] Geijtenbeek, T.B.; Krooshoop, D.J.; Bleijs, D.A.; van Vliet, S.J.; van Duijnhoven, G.C.; Grabovsky, V.; Alon, R.; Figdor, C.G.; van
Kooyk, Y. DC-SIGN-ICAM-2 interaction mediates dendritic cell trafficking. Nat. Immunol. 2000, 1, 353–357. [CrossRef]
57. Kwon, D.S.; Gregorio, G.; Bitton, N.; Hendrickson, W.A.; Littman, D.R. DC-SIGN-mediated internalization of HIV is required for
trans-enhancement of T cell infection. Immunity 2002, 16, 135–144. [CrossRef] y
g
[
]
57. Kwon, D.S.; Gregorio, G.; Bitton, N.; Hendrickson, W.A.; Littman, D.R. DC-SIGN-mediated internalization of HIV is required for
trans-enhancement of T cell infection. Immunity 2002, 16, 135–144. [CrossRef] y
58. McDonald, D.; Wu, L.; Bohks, S.M.; KewalRamani, V.N.; Unutmaz, D.; Hope, T.J. Recruitment of HIV and its receptors to dendritic
cell-T cell junctions. Science 2003, 300, 1295–1297. [CrossRef] j
59. Cameron, P.U.; Freudenthal, P.S.; Barker, J.M.; Gezelter, S.; Inaba, K.; Steinman, R.M. Dendritic cells exposed to human
immunodeficiency virus type-1 transmit a vigorous cytopathic infection to CD4+ T cells. Science 1992, 257, 383–387. [CrossRef] j
59. Cameron, P.U.; Freudenthal, P.S.; Barker, J.M.; Gezelter, S.; Inaba, K.; Steinman, R.M. Dendritic cells exposed to human
immunodeficiency virus type-1 transmit a vigorous cytopathic infection to CD4+ T cells. Science 1992, 257, 383–387. [CrossRef]
60. Cavrois, M.; Neidleman, J.; Greene, W.C. The achilles heel of the trojan horse model of HIV-1 trans-infection. PLoS Pathog. 2008,
4, e1000051. [CrossRef] 60. Cavrois, M.; Neidleman, J.; Greene, W.C. The achilles heel of the trojan horse model of HIV-1 trans-inf
4, e1000051. [CrossRef] 61. Boukour, S.; Masse, J.M.; Benit, L.; Dubart-Kupperschmitt, A.; Cramer, E.M. Lentivirus degradation and DC-SIGN expression by
human platelets and megakaryocytes. J. Thrombosis Haemost. JTH 2006, 4, 426–435. [CrossRef] [PubMed] 62. Chaipan, C.; Soilleux, E.J.; Simpson, P.; Hofmann, H.; Gramberg, T.; Marzi, A.; Geier, M.; Stewart, E.A.; Eisemann, J.; Steinkasserer,
A.; et al. DC-SIGN and CLEC-2 mediate human immunodeficiency virus type 1 capture by platelets. J. Virol. 2006, 80, 8951–8960. [CrossRef] [PubMed] 63. Rappocciolo, G.; Piazza, P.; Fuller, C.L.; Reinhart, T.A.; Watkins, S.C.; Rowe, D.T.; Jais, M.; Gupta, P.; Rinaldo, C.R. DC-SIGN on B
lymphocytes is required for transmission of HIV-1 to T lymphocytes. PLoS Pathog. 2006, 2, e70. [CrossRef] [PubMed] 64. Thépaut, M.; Luczkowiak, J.; Vivès, C.; Labiod, N.; Bally, I.; Lasala, F.; Grimoire, Y.; Fenel, D.; Sattin, S.; Thielens, N.; et al. DC/L-SIGN recognition of spike glycoprotein promotes SARS-CoV-2 trans-infection and can be inhibited by a glycomimetic
antagonist. bioRxiv 2020. [CrossRef] 65. 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] Identification of four novel DC-SIGN ligands on
Mycobacterium bovis BCG. Protein Cell 2010, 1, 859–870. [CrossRef] y
47. Halary, F.; Amara, A.; Lortat-Jacob, H.; Messerle, M.; Delaunay, T.; Houles, C.; Fieschi, F.; Arenzana-Seisdedos, F.; Moreau,
J.F.; Dechanet-Merville, J. Human cytomegalovirus binding to DC-SIGN is required for dendritic cell infection and target cell
trans-infection. Immunity 2002, 17, 653–664. [CrossRef] y
48. Gramberg, T.; Hofmann, H.; Moller, P.; Lalor, P.F.; Marzi, A.; Geier, M.; Krumbiegel, M.; Winkler, T.; Kirchhoff, F.; Adams,
D.H.; et al. LSECtin interacts with filovirus glycoproteins and the spike protein of SARS coronavirus. Virology 2005, 340, 224–236. [CrossRef] [
]
49. Shimojima, M.; Stroher, U.; Ebihara, H.; Feldmann, H.; Kawaoka, Y. Identification of cell surface molecules involved in
dystroglycan-independent Lassa virus cell entry. J. Virol. 2012, 86, 2067–2078. [CrossRef] 50. Geijtenbeek, T.B.H.; Engering, A.; Van Kooyk, Y. DC-SIGN, a C-type lectin on dendritic cells that unveils many aspects of dendritic
cell biology. J. Leukoc. Biol. 2002, 71, 921–931. 51. Geijtenbeek, T.B.; Kwon, D.S.; Torensma, R.; van Vliet, S.J.; van Duijnhoven, G.C.; Middel, J.; Cornelissen, I.L.; Nottet, H.S.;
KewalRamani, V.N.; Littman, D.R.; et al. DC-SIGN, a dendritic cell-specific HIV-1-binding protein that enhances trans-infection
of T cells. Cell 2000, 100, 587–597. [CrossRef] 52. Moris, A.; Nobile, C.; Buseyne, F.; Porrot, F.; Abastado, J.-P.; Schwartz, O. DC-SIGN promotes exogenous MHC-I-restricted HIV-1
antigen presentation. Blood 2004, 103, 2648–2654. [CrossRef] [PubMed] 53. Turville, S.G.; Santos, J.J.; Frank, I.; Cameron, P.U.; Wilkinson, J.; Miranda-Saksena, M.; Dable, J.; Stossel, H.; Romani, N.; Piatak,
M., Jr.; et al. Immunodeficiency virus uptake, turnover, and 2-phase transfer in human dendritic cells. Blood 2004, 103, 2170–2179. [CrossRef] [PubMed] 54. Hijazi, K.; Wang, Y.; Scala, C.; Jeffs, S.; Longstaff, C.; Stieh, D.; Haggarty, B.; Vanham, G.; Schols, D.; Balzarini, J.; et al. DC-SIGN
increases the affinity of HIV-1 envelope glycoprotein interaction with CD4. PLoS ONE 2011, 6, e28307. [CrossRef] 55. Burleigh, L.; Lozach, P.-Y.; Schiffer, C.; Staropoli, I.; Pezo, V.; Porrot, F.; Canque, B.; Virelizier, J.-L.; Arenzana-Seisdedos, F.; Amara,
A. Infection of dendritic cells (DCs), not DC-SIGN-mediated internalization of human immunodeficiency virus, is required for
long-term transfer of virus to T cells. J. Virol. 2006, 80, 2949–2957. [CrossRef] g
56. Geijtenbeek, T.B.; Krooshoop, D.J.; Bleijs, D.A.; van Vliet, S.J.; van Duijnhoven, G.C.; Grabovsky, V.; Alon, R.; Figdor, C.G.; van
Kooyk, Y. DC-SIGN-ICAM-2 interaction mediates dendritic cell trafficking. Nat. Immunol. 2000, 1, 353–357. [CrossRef] g
56. 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] Lin, G.; Simmons, G.; Pohlmann, S.; Baribaud, F.; Ni, H.; Leslie, G.J.; Haggarty, B.S.; Bates, P.; Weissman, D.; Hoxie, J.A.; et al. Differential N-linked glycosylation of human immunodeficiency virus and Ebola virus envelope glycoproteins modulates
interactions with DC-SIGN and DC-SIGNR. J. Virol. 2003, 77, 1337–1346. [CrossRef] J
38. Lai, W.K.; Sun, P.J.; Zhang, J.; Jennings, A.; Lalor, P.F.; Hubscher, S.; McKeating, J.A.; Adams, D.H. Expression of DC-SIGN
and DC-SIGNR on human sinusoidal endothelium: A role for capturing hepatitis C virus particles. Am. J. Pathol. 2006, 169,
200–208. [CrossRef] 39. de Witte, L.; Abt, M.; Schneider-Schaulies, S.; van Kooyk, Y.; Geijtenbeek, T.B.H. Measles virus targets DC-SIGN to enhance
dendritic cell infection. J. Virol. 2006, 80, 3477–3486. [CrossRef] 40. de Jong, M.A.W.P.; de Witte, L.; Bolmstedt, A.; van Kooyk, Y.; Geijtenbeek, T.B.H. Dendritic cells med
infection and transmission through the C-type lectin DC-SIGN. J. Gen. Virol. 2008, 89, 2398–2409. [Cros 41. Mou, H.; Raj, V.S.; van Kuppeveld, F.J.M.; Rottier, P.J.M.; Haagmans, B.L.; Bosch, B.J. The receptor binding domain of the new
Middle East respiratory syndrome coronavirus maps to a 231-residue region in the spike protein that efficiently elicits neutralizing
antibodies. J. Virol. 2013, 87, 9379–9383. [CrossRef] [PubMed] 42. Goncalves, A.-R.; Moraz, M.-L.; Pasquato, A.; Helenius, A.; Lozach, P.-Y.; Kunz, S. Role of DC-SIG
human dendritic cells. J. Virol. 2013, 87, 11504–11515. [CrossRef] [PubMed] 43. Johnson, T.R.; McLellan, J.S.; Graham, B.S. Respiratory syncytial virus glycoprotein G interacts with DC-SIGN and L-SIGN to
activate ERK1 and ERK2. J. Virol. 2012, 86, 1339–1347. [CrossRef] activate ERK1 and ERK2. J. Virol. 2012, 86, 1339–1347. [CrossRef]
44. Lozach, P.-Y.; Kuhbacher, A.; Meier, R.; Mancini, R.; Bitto, D.; Bouloy, M.; Helenius, A. DC-SIGN as a receptor for phleboviruses. Cell Host Microbe 2011, 10, 75–88. [CrossRef] [PubMed] 44. Lozach, P.-Y.; Kuhbacher, A.; Meier, R.; Mancini, R.; Bitto, D.; Bouloy, M.; Helenius, A. DC-SIGN as a receptor for phleboviruses. Cell Host Microbe 2011, 10, 75–88. [CrossRef] [PubMed] 44. Lozach, P.-Y.; Kuhbacher, A.; Meier, R.; Mancini, R.; Bitto, D.; B
Cell Host Microbe 2011, 10, 75–88. [CrossRef] [PubMed] 45. Davis, C.W.; Nguyen, H.-Y.; Hanna, S.L.; Sanchez, M.D.; Doms, R.W.; Pierson, T.C. West Nile virus discriminates between
DC-SIGN and DC-SIGNR for cellular attachment and infection. J. Virol. 2006, 80, 1290–1301. [CrossRef] 12 of 15 12 of 15 Biology 2021, 10, 1 46. Carroll, M.V.; Sim, R.B.; Bigi, F.; Jakel, A.; Antrobus, R.; Mitchell, D.A. 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] Hodges, A.; Sharrocks, K.; Edelmann, M.; Baban, D.; Moris, A.; Schwartz, O.; Drakesmith, H.; Davies, K.; Kessler, B.; McMichael,
A.; et al. Activation of the lectin DC-SIGN induces an immature dendritic cell phenotype triggering Rho-GTPase activity required
for HIV-1 replication. Nat. Immunol. 2007, 8, 569–577. [CrossRef] p
66. Gringhuis, S.I.; den Dunnen, J.; Litjens, M.; van Het Hof, B.; van Kooyk, Y.; Geijtenbeek, T.B.H. C-type lectin DC-SIGN modulates
Toll-like receptor signaling via Raf-1 kinase-dependent acetylation of transcription factor NF-kappaB. Immunity 2007, 26, 605–616. [CrossRef] 67. Geijtenbeek, T.B.H.; Gringhuis, S.I. Signalling through C-type lectin receptors: Shaping immune responses. Nat. Rev. Immunol. 2009, 9, 465–479. [CrossRef] 68. Delmas, B.; Gelfi, J.; L’Haridon, R.; Vogel, L.K.; Sjostrom, H.; Noren, O.; Laude, H. Aminopeptidase N is a major receptor for
the entero-pathogenic coronavirus TGEV. Nature 1992, 357, 417–420. [CrossRef] 69. Yeager, C.L.; Ashmun, R.A.; Williams, R.K.; Cardellichio, C.B.; Shapiro, L.H.; Look, A.T.; Holmes, K.V. H
is a receptor for human coronavirus 229E. Nature 1992, 357, 420–422. [CrossRef] p
70. Matrosovich, M.; Herrler, G.; Klenk, H.D. Sialic Acid Receptors of Viruses. Top Curr. Chem. 2015, 367, 1–28. 13 of 15 13 of 15 Biology 2021, 10, 1 71. Huang, X.; Dong, W.; Milewska, A.; Golda, A.; Qi, Y.; Zhu, Q.K.; Marasco, W.A.; Baric, R.S.; Sims, A.C.; Pyrc, K.; et al. Human Coronavirus HKU1 Spike Protein Uses O-Acetylated Sialic Acid as an Attachment Receptor Determinant and Employs
H
l ti i
E t
P
t i
R
t
D
t
i
E
J Vi
l 2015 89 7202 7213 [C
R f] [P bM d] gg
p
y
g
y
72. Raj, V.S.; Mou, H.; Smits, S.L.; Dekkers, D.H.W.; Muller, M.A.; Dijkman, R.; Muth, D.; Demmers, J.A.A.; Zaki, A.; Fouchier,
R.A.M.; et al. Dipeptidyl peptidase 4 is a functional receptor for the emerging human coronavirus-EMC. Nature 2013, 495, 251–254. [CrossRef] [PubMed] 73. Chan, C.-M.; Chu, H.; Wang, Y.; Wong, B.H.-Y.; Zhao, X.; Zhou, J.; Yang, D.; Leung, S.P.; Chan, J.F.-W.; Yeung, M.-L.; et al. Carcinoembryonic Antigen-Related Cell Adhesion Molecule 5 Is an Important Surface Attachment Factor That Facilitates Entry
of Middle East Respiratory Syndrome Coronavirus. J. Virol. 2016, 90, 9114–9127. [CrossRef] [PubMed] p
y y
74. Hoffmann, M.; Kleine-Weber, H.; Schroeder, S.; Kruger, N.; Herrler, T.; Erichsen, S.; Schiergens, T.S.; Herrler, G.; Wu, N.H.; Nitsche,
A.; et al. SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor. 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] Cell
2020, 181, 271–280. [CrossRef] 75. Hofmann, H.; Pyrc, K.; van der Hoek, L.; Geier, M.; Berkhout, B.; Pohlmann, S. Human coronavirus NL63 employs the severe
acute respiratory syndrome coronavirus receptor for cellular entry. Proc. Natl. Acad. Sci. USA 2005, 102, 7988–7993. [CrossRef] 76. Li, W.; Moore, M.J.; Vasilieva, N.; Sui, J.; Wong, S.K.; Berne, M.A.; Somasundaran, M.; Sullivan, J.L.; Luzuriaga, K.; Greenough,
T.C.; et al. Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus. Nature 2003, 426, 450–454. [CrossRef] [
]
77. Han, D.P.; Lohani, M.; Cho, M.W. Specific asparagine-linked glycosylation sites are critical for DC-SIGN- and L-SIGN-mediated
severe acute respiratory syndrome coronavirus entry. J. Virol. 2007, 81, 12029–12039. [CrossRef] 78. Soh, W.T.; Liu, Y.; Nakayama, E.E.; Ono, C.; Torii, S.; Nakagami, H.; Matsuura, Y.; Shioda, T.; Arase, H. The N-terminal domain of
spike glycoprotein mediates SARS-CoV-2 infection by associating with L-SIGN and DC-SIGN. bioRxiv 2020. [CrossRef] y
g
spike glycoprotein mediates SARS-CoV-2 infection by associating with L-SIGN and DC-SIGN. bioRxiv 2020. [CrossRef] 79. Gao, C.; Zeng, J.; Jia, N.; Stavenhagen, K.; Matsumoto, Y.; Zhang, H.; Li, J.; Hume, A.J.; Muhlber
SARS-CoV-2 Spike Protein Interacts with Multiple Innate Immune Receptors. bioRxiv 2020. [CrossRef] 79. Gao, C.; Zeng, J.; Jia, N.; Stavenhagen, K.; Matsumoto, Y.; Zhang, H.; Li, J.; Hume, A.J.; Muhlberger, E.; van Die, I.; et al. SARS-CoV-2 Spike Protein Interacts with Multiple Innate Immune Receptors. bioRxiv 2020. [CrossRef] 80. Chiodo, F.; Bruijns, S.C.M.; Rodriguez, E.; Li, R.J.E.; Molinaro, A.; Silipo, A.; Di Lorenzo, F.; Garcia-Rivera, D.; Valdes-Balbin,
Y.; Verez-Bencomo, V.; et al. Novel ACE2-Independent Carbohydrate-Binding of SARS-CoV-2 Spike Protein to Host Lectins
and Lung Microbiota. bioRxiv 2020. [CrossRef] 81. Snyder, G.A.; Colonna, M.; Sun, P.D. The structure of DC-SIGNR with a portion of its repeat domain lends insights to modeling
of the receptor tetramer. J. Mol. Biol. 2005, 347, 979–989. [CrossRef] [PubMed] 82. Sol-Foulon, N.; Moris, A.; Nobile, C.; Boccaccio, C.; Engering, A.; Abastado, J.P.; Heard, J.M.; van Kooyk, Y.; Schwartz, O. HIV-1
Nef-induced upregulation of DC-SIGN in dendritic cells promotes lymphocyte clustering and viral spread. Immunity 2002, 16,
145–155. [CrossRef] 83. Mitchell, D.A.; Fadden, A.J.; Drickamer, K. A novel mechanism of carbohydrate recognition by the C-type lectins DC-SIGN
and DC-SIGNR. Subunit organization and binding to multivalent ligands. J. Biol. Chem. 2001, 276, 28939–28945. [CrossRef]
[PubMed] [
]
84. 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] Cambi, A.; de Lange, F.; van Maarseveen, N.M.; Nijhuis, M.; Joosten, B.; van Dijk, E.M.; de Bakker, B.I.; Fransen, J.A.; Bovee-Geurts,
P.H.; van Leeuwen, F.N.; et al. Microdomains of the C-type lectin DC-SIGN are portals for virus entry into dendritic cells. J. Cell
Biol. 2004, 164, 145–155. [CrossRef] [PubMed] ,
,
[
] [
]
85. Yu, Q.D.; Oldring, A.P.; Powlesland, A.S.; Tso, C.K.; Yang, C.; Drickamer, K.; Taylor, M.E. Autonomous tetramerization domains
in the glycan-binding receptors DC-SIGN and DC-SIGNR. J. Mol. Biol. 2009, 387, 1075–1080. [CrossRef] [PubMed] 86. Feinberg, H.; Tso, C.K.; Taylor, M.E.; Drickamer, K.; Weis, W.I. Segmented helical structure of the neck region of the glycan-binding
receptor DC-SIGNR. J. Mol. Biol. 2009, 394, 613–620. [CrossRef] [PubMed] 86. Feinberg, H.; Tso, C.K.; Taylor, M.E.; Drickamer, K.; Weis, W.I. Segmented helical structure of t
receptor DC-SIGNR. J. Mol. Biol. 2009, 394, 613–620. [CrossRef] [PubMed] Feinberg, H.; Tso, C.K.; Taylor, M.E.; Drickamer, K.; Weis, W.I. Segmented helical structure of the neck regio
receptor DC-SIGNR. J. Mol. Biol. 2009, 394, 613–620. [CrossRef] [PubMed] p
J
87. Drickamer, K. C-type lectin-like domains. Curr. Opin. Struct. Biol. 1999, 9, 585–590. [CrossRef] yp
p
88. Van Liempt, E.; Imberty, A.; Bank, C.M.; Van Vliet, S.J.; Van Kooyk, Y.; Geijtenbeek, T.B.; Van Die, I. Molecular basis of
the differences in binding properties of the highly related C-type lectins DC-SIGN and L-SIGN to Lewis X trisaccharide
and Schistosoma mansoni egg antigens. J. Biol. Chem. 2004, 279, 33161–33167. [CrossRef] 89. Watanabe, Y.; Allen, J.D.; Wrapp, D.; McLellan, J.S.; Crispin, M. Site-specific glycan analysis of th
2020, 369, 330–333. [CrossRef] J.D.; Wrapp, D.; McLellan, J.S.; Crispin, M. Site-specific glycan analysis of the SARS-CoV-2 spike. Science
CrossRef] 90. Chandler, K.B.; Leon, D.R.; Kuang, J.; Meyer, R.D.; Rahimi, N.; Costello, C.E. N-Glycosylation regulates ligand-dependent
activation and signaling of vascular endothelial growth factor receptor 2 (VEGFR2). J. Biol. Chem. 2019, 294, 13117–13130. [CrossRef] 91. Daeron, M.; Jaeger, S.; Du Pasquier, L.; Vivier, E. Immunoreceptor tyrosine-based inhibition motifs: A qu
Immunol. Rev. 2008, 224, 11–43. [CrossRef] [PubMed] 92. Rahimi, N. The ubiquitin-proteasome system meets angiogenesis. Mol. Cancer Ther. 2012, 11, 538–548. [CrossRef] [PubMed] 92. Rahimi, N. The ubiquitin-proteasome system meets angiogenesis. Mol. Cancer Ther. 2012, 11, 538–548. [CrossRef] [PubMed]
93. Liu, H.; Zhu, T. Determination of DC-SIGN and DC-SIGNR repeat region variations. Methods Mol. Biol. 2005, 304, 471–481. [CrossRef] [PubMed] 93. Liu, H.; Zhu, T. 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] Chan, V.S.; Chan, K.Y.; Chen, Y.; Poon, L.L.; Cheung, A.N.; Zheng, B.; Chan, K.H.; Mak, W.; Ngan, H.Y.; Xu, X.; et al. Homozygous polymorphism of DC SIGNR in peri centenarian from Han Chinese population. BMC Med. Genet. 2009, 10, 134. [CrossRef]
108. Chan, V.S.; Chan, K.Y.; Chen, Y.; Poon, L.L.; Cheung, A.N.; Zheng, B.; Chan, K.H.; Mak, W.; Ngan, H.Y.; Xu, X.; et al. Homozygous
L SIGN (CLEC4M)
l
t
ti
l i
SARS
i
i f
ti
N t G
t 2006 38 38 46 [C
R f] p
y
p
p
p p
108. Chan, V.S.; Chan, K.Y.; Chen, Y.; Poon, L.L.; Cheung, A.N.; Zheng, B.; Chan, K.H.; Mak, W.; Ngan, H.Y.; Xu, X.; et al. Homozygous
L-SIGN (CLEC4M) plays a protective role in SARS coronavirus infection. Nat. Genet. 2006, 38, 38–46. [CrossRef] L-SIGN (CLEC4M) plays a protective role in SARS coronavirus infection. Nat. Genet. 2006, 38, 38–46. [CrossRef]
109. Fawcett, J.; Holness, C.L.; Needham, L.A.; Turley, H.; Gatter, K.C.; Mason, D.Y.; Simmons, D.L. Molecular cloning of ICAM-3, (
) p y
p
,
,
[
]
109. Fawcett, J.; Holness, C.L.; Needham, L.A.; Turley, H.; Gatter, K.C.; Mason, D.Y.; Simmons, D.L. Molecular cloning of ICAM-3, (
) p y
p
109. Fawcett, J.; Holness, C.L.; Needham, L.A.; Turley, H.; Gatter, K.C.; Mason, D.Y.; Simmons, D.L. Molecular cloning of ICAM-3,
a third ligand for LFA-1, constitutively expressed on resting leukocytes. Nature 1992, 360, 481–484. [CrossRef] 109. Fawcett, J.; Holness, C.L.; Needham, L.A.; Turley, H.; Gatter, K.C.; Mason, D.Y.; Simmons, D.L. Mole
a third ligand for LFA-1, constitutively expressed on resting leukocytes. Nature 1992, 360, 481–484. [Cr g
y
p
g
y
110. Kristof, E.; Zahuczky, G.; Katona, K.; Doro, Z.; Nagy, E.; Fesus, L. Novel role of ICAM3 and LFA-1 in the clearance of apoptotic
neutrophils by human macrophages. Apoptosis 2013, 18, 1235–1251. [CrossRef] 110. Kristof, E.; Zahuczky, G.; Katona, K.; Doro, Z.; Nagy, E.; Fesus, L. Novel role of ICAM3 and
neutrophils by human macrophages. Apoptosis 2013, 18, 1235–1251. [CrossRef] 110. Kristof, E.; Zahuczky, G.; Katona, K.; Doro, Z.; Nagy, E.; Fesus, L. Novel role of ICAM3 and LFA-1 in the clearance of apoptotic
neutrophils by human macrophages. Apoptosis 2013, 18, 1235–1251. [CrossRef] ,
;
y,
;
,
;
,
;
gy,
;
,
utrophils by human macrophages. Apoptosis 2013, 18, 1235–1251. [CrossRef] p
y
p
g
p p
111. 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] Kuba, K.; Imai, Y.; Rao, S.; Gao, H.; Guo, F.; Guan, B.; Huan, Y.; Yang, P.; Zhang, Y.; Deng, W.; et al. A crucial role of angiotensin
converting enzyme 2 (ACE2) in SARS coronavirus-induced lung injury. Nat. Med. 2005, 11, 875–879. [CrossRef] [PubMed] g
y
g
j
y
102. Zhou, P.; Yang, X.L.; Wang, X.G.; Hu, B.; Zhang, L.; Zhang, W.; Si, H.R.; Zhu, Y.; Li, B.; Huang, C.L.; et al. A pneumonia outbreak
associated with a new coronavirus of probable bat origin. Nature 2020, 579, 270–273. [CrossRef] [PubMed] 102. Zhou, P.; Yang, X.L.; Wang, X.G.; Hu, B.; Zhang, L.; Zhang, W.; Si, H.R.; Zhu, Y.; Li, B.; Huang, C.L.; et al. A pneumonia outbreak
associated with a new coronavirus of probable bat origin. Nature 2020, 579, 270–273. [CrossRef] [PubMed] 103. Hamming, I.; Timens, W.; Bulthuis, M.L.; Lely, A.T.; Navis, G.; van Goor, H. Tissue distribution of ACE2 protein, the functional
receptor for SARS coronavirus. A first step in understanding SARS pathogenesis. J. Pathol. 2004, 203, 631–637. [CrossRef]
[PubMed] 104. Hikmet, F.; Méar, L.; Edvinsson, Å.; Micke, P.; Uhlén, M.; Lindskog, C. The protein expression profile of ACE2 in human tissues. bioRxiv 2020. [CrossRef] [PubMed] 105. Ganier, C.; Du-Harpur, X.; Harun, N.; Wan, B.; Arthurs, C.; Luscombe, N.; Watt, F.; Lynch, M. CD147 (BSG) but not ACE2
expression is detectable in vascular endothelial cells within single cell RNA sequencing datasets derived from multiple tissues in
healthy individuals. bioRxiv 2020. [CrossRef] y
106. Engering, A.; van Vliet, S.J.; Hebeda, K.; Jackson, D.G.; Prevo, R.; Singh, S.K.; Geijtenbeek, T.B.; van Krieken, H.; van Kooyk, Y. Dynamic populations of dendritic cell-specific ICAM-3 grabbing nonintegrin-positive immature dendritic cells and liver/lymph
node-specific ICAM-3 grabbing nonintegrin-positive endothelial cells in the outer zones of the paracortex of human lymph nodes. Am. J. Pathol. 2004, 164, 1587–1595. [CrossRef] Wang, J.X.; Tang, N.L.; Xie, L.; Gong, Y.Y.; Yang, Z.; Xu, L.Y.; Kong, Q.P.; Zhang, Y.P. The neck-region
C-SIGNR in peri-centenarian from Han Chinese population. BMC Med. Genet. 2009, 10, 134. [CrossRef] 107. Li, H.; Wang, C.Y.; Wang, J.X.; Tang, N.L.; Xie, L.; Gong, Y.Y.; Yang, Z.; Xu, L.Y.; Kong, Q.P.; Z
polymorphism of DC-SIGNR in peri-centenarian from Han Chinese population. BMC Med. Genet. polymorphism of DC-SIGNR in peri-centenarian from Han Chinese population. BMC Med. Genet. 2009, 10, 134. [CrossRef]
108. 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] Determination of DC-SIGN and DC-SIGNR repeat region variations. Methods Mol. Biol. 2005, 304, 471–481. [CrossRef] [PubMed] 94. Zhu, D.; Kawana-Tachikawa, A.; Iwamoto, A.; Kitamura, Y. Influence of polymorphism in dendritic cell-specific intercellular
adhesion molecule-3-grabbing nonintegrin-related (DC-SIGNR) gene on HIV-1 trans-infection. Biochem. Biophys. Res. Commun. 2010, 393, 598–602. [CrossRef] [PubMed] 14 of 15 14 of 15 Biology 2021, 10, 1 95. Kashima, S.; Rodrigues, E.S.; Azevedo, R.; da Cruz Castelli, E.; Mendes-Junior, C.T.; Yoshioka, F.K.N.; da Silva, I.T.; Takayanagui,
O.M.; Covas, D.T. DC-SIGN (CD209) gene promoter polymorphisms in a Brazilian population and their association with human
T-cell lymphotropic virus type 1 infection. J. Gen. Virol. 2009, 90, 927–934. [CrossRef] [PubMed] 96. Barreiro, L.B.; Neyrolles, O.; Babb, C.L.; Tailleux, L.; Quach, H.; McElreavey, K.; Helden, P.D.; Hoal, E.G.; Gicquel, B.; Quintana-
Murci, L. Promoter variation in the DC-SIGN-encoding gene CD209 is associated with tuberculosis. PLoS Med. 2006, 3, e20. [CrossRef] 97. Kagone, T.S.; Bisseye, C.; Meda, N.; Testa, J.; Pietra, V.; Kania, D.; Yonli, A.T.; Compaore, T.R.; Nikiema, J.B.; de Souza, C.; et al. A variant of DC-SIGN gene promoter associated with resistance to HIV-1 in serodiscordant couples in Burkina Faso. Asian Pac. J. Trop. Med. 2014, 7S1, S93–S96. [CrossRef] p
,
,
[
]
98. Tailleux, L.; Pham-Thi, N.; Bergeron-Lafaurie, A.; Herrmann, J.-L.; Charles, P.; Schwartz, O.; Scheinmann, P.; Lagrange, P.H.;
de Blic, J.; Tazi, A.; et al. DC-SIGN induction in alveolar macrophages defines privileged target host cells for mycobacteria in
patients with tuberculosis. PLoS Med. 2005, 2, e381. [CrossRef] p
[
]
99. Pohlmann, S.; Soilleux, E.J.; Baribaud, F.; Leslie, G.J.; Morris, L.S.; Trowsdale, J.; Lee, B.; Coleman, N.; Doms, R.W. DC-SIGNR,
a DC-SIGN homologue expressed in endothelial cells, binds to human and simian immunodeficiency viruses and activates
infection in trans. Proc. Natl. Acad. Sci. USA 2001, 98, 2670–2675. [CrossRef] 100. Uhlen, M.; Fagerberg, L.; Hallstrom, B.M.; Lindskog, C.; Oksvold, P.; Mardinoglu, A.; Sivertsson, A.; Kampf, C.; Sjostedt, E.;
Asplund, A.; et al. Proteomics. Tissue-based map of the human proteome. Science 2015, 347, 1260419. [CrossRef] p
p
p
101. Kuba, K.; Imai, Y.; Rao, S.; Gao, H.; Guo, F.; Guan, B.; Huan, Y.; Yang, P.; Zhang, Y.; Deng, W.; et al. A crucial role of angiotensin
converting enzyme 2 (ACE2) in SARS coronavirus-induced lung injury. Nat. Med. 2005, 11, 875–879. [CrossRef] [PubMed] 101. 119. Xiang-Hua, Y.; Le-Min, W.; Ai-Bin, L.; Zhu, G.; Riquan, L.; Xu-You, Z.; Wei-Wei, R.; Ye-Nan, W. Severe acute respiratory syndrome
and venous thromboembolism in multiple organs. Am. J. Respir. Crit. Care Med. 2010, 182, 436–437. [CrossRef] p
g
J
p
[
]
120. Ng, K.H.; Wu, A.K.; Cheng, V.C.; Tang, B.S.; Chan, C.Y.; Yung, C.Y.; Luk, S.H.; Lee, T.W.; Chow, L.; Yuen, K.Y. Pulmonary artery
thrombosis in a patient with severe acute respiratory syndrome. Postgrad. Med. J. 2005, 81, e3. [CrossRef] p
p
y y
g
121. Zhou, F.; Yu, T.; Du, R.; Fan, G.; Liu, Y.; Liu, Z.; Xiang, J.; Wang, Y.; Song, B.; Gu, X.; et al. Clinical course and risk factors
for mortality of adult inpatients with COVID-19 in Wuhan, China: A retrospective cohort study. Lancet 2020, 395, 1054–1062.
[CrossRef] 1.
Mulloy, B.; Linhardt, R.J. Order out of complexity—Protein structures that interact with heparin. Curr. Opin. Struct. Biol. 2001, 11,
623–628. [CrossRef] Patey, N.; Vazeux, R.; Canioni, D.; Potter, T.; Gallatin, W.M.; Brousse, N. Intercellular adhesion molecule-3 on endothelial cells. Expression in tumors but not in inflammatory responses. Am. J. Pathol. 1996, 148, 465–472. [PubMed] 112. Huang, M.T.; Mason, J.C.; Birdsey, G.M.; Amsellem, V.; Gerwin, N.; Haskard, D.O.; Ridley, A.J.; Randi, A.M. Endothelial
intercellular adhesion molecule (ICAM)-2 regulates angiogenesis. Blood 2005, 106, 1636–1643. [CrossRef] [PubMed] 113. O’Sullivan, J.M.; Gonagle, D.M.; Ward, S.E.; Preston, R.J.S.; O’Donnell, J.S. Endothelial cells orchestr
Lancet Haematol. 2020, 7, e553–e555. [CrossRef] nagle, D.M.; Ward, S.E.; Preston, R.J.S.; O’Donnell, J.S. Endothelial cells orchestrate COVID-19 coagulopathy
0, 7, e553–e555. [CrossRef] 114. Ackermann, M.; Verleden, S.E.; Kuehnel, M.; Haverich, A.; Welte, T.; Laenger, F.; Vanstapel, A.; Werlein, C.; Stark, H.; Tzankov,
A.; et al. Pulmonary Vascular Endothelialitis, Thrombosis, and Angiogenesis in Covid-19. N. Engl. J. Med. 2020, 383, 120–128. [CrossRef] [PubMed] 115. Wiersinga, W.J.; Rhodes, A.; Cheng, A.C.; Peacock, S.J.; Prescott, H.C. Pathophysiology, Transmissio
of Coronavirus Disease 2019 (COVID-19): A Review. JAMA 2020, 324, 782–793. [CrossRef] 116. Fried, J.A.; Ramasubbu, K.; Bhatt, R.; Topkara, V.K.; Clerkin, K.J.; Horn, E.; Rabbani, L.; Brodie, D.; Jain, S.S.; Kirtane, A.J.; et al. The Variety of Cardiovascular Presentations of COVID-19. Circulation 2020, 141, 1930–1936. [CrossRef] y
117. Arentz, M.; Yim, E.; Klaff, L.; Lokhandwala, S.; Riedo, F.X.; Chong, M.; Lee, M. Characteristics and Outcomes of 21 Critically Ill
P
i
Wi h COVID 19 i
W
hi
S
JAMA 2020 323 1612 1614 [C
R f] y
117. Arentz, M.; Yim, E.; Klaff, L.; Lokhandwala, S.; Riedo, F.X.; Chong, M.; Lee, M. Charact
Patients With COVID-19 in Washington State. JAMA 2020, 323, 1612–1614. [CrossRef] y
117. Arentz, M.; Yim, E.; Klaff, L.; Lokhandwala, S.; Riedo, F.X.; Chong, M.; Lee, M. Characteristics and Outcomes of 21 Critically Ill
Patients With COVID-19 in Washington State. JAMA 2020, 323, 1612–1614. [CrossRef] 118. Richardson, S.; Hirsch, J.S.; Narasimhan, M.; Crawford, J.M.; McGinn, T.; Davidson, K.W.; the Northwell, C.-R.C.; Barnaby, D.P.;
Becker, L.B.; Chelico, J.D.; et al. Presenting Characteristics, Comorbidities, and Outcomes Among 5700 Patients Hospitalized
With COVID-19 in the New York City Area. JAMA 2020, 323, 2052–2059. [CrossRef] 15 of 15 15 of 15 Biology 2021, 10, 1
|
https://openalex.org/W4286895527
|
https://zenodo.org/records/6299984/files/Paper%205%20-%20Prosiding%20AW9.pdf
|
English
| null |
Traffic Scofflaw and Electronic Ticket Effectiveness Model
|
Zenodo (CERN European Organization for Nuclear Research)
| 2,021
|
cc-by
| 3,702
|
“SMART CITY AS AN ALTERNATIVE SOLUTION
FOR URBAN DEVELOPMENT AND EQUITY IN
INDONESIA”
p-ISSN 2477-3026
e-ISSN 2746-4865
Vol. 9, No. 1, 2022
9 “SMART CITY AS AN ALTERNATIVE SOLUTION
FOR URBAN DEVELOPMENT AND EQUITY IN
INDONESIA”
p-ISSN 2477-3026
e-ISSN 2746-4865
Vol. 9, No. 1, 2022
9 9 9 “SMART CITY AS AN ALTERNATIVE SOLUTION
FOR URBAN DEVELOPMENT AND EQUITY IN
INDONESIA”
Dilakukan secara daring, 23-24 Oktober 2021 “SMART CITY AS AN ALTERNATIVE SOLUTION
FOR URBAN DEVELOPMENT AND EQUITY IN
INDONESIA” Dilakukan secara daring, 23-24 Oktober 2021 a) putri.25117066@student.itera.ac.id, b)tastaptyani.kurnia@tl.itera.ac.id,
c)mutiara.fajar@tl.itera.ac.id Abstract. Today the urban cities are facing increased water stress especially for available water
resources. Water resources is not only used for domestic purposes, but also for non-domestic
purposes or public facilities. One example of a public facility that uses water to meet daily
needs is a mosque. In urban infrastructure has mosque as moeslem worship places and the
building need to be environmentally sustainable. Rainwater harvesting system is one method
for water conservation and relate for sustainable infrastructure. Rainwater harvesting has a pre-
eminent role to play and utilization of rainwater and also to reduce run off. By looking at this
condition, rainwater should be utilized optimally. This aim study for planning of rainwater
harvesting system at mosque with the main purpose of this planning being as an alternative to
clean water and minimizing the use of ground water for water conservation. The data needed in
planning the rainwater harvesting system are data on water demand, rainfall, and as built
drawings. After the calculation, the components of the rainwater harvesting system design are
obtained, namely storage tanks (3,100 L), gutters (6 inches), first flush diverter (4 inches with a
length of 7 meters), and piping systems (4 inches). The construction of this rainwater harvesting
system is expected as an alternative to minimize the use of ground water and clean water
supplies in the future, especially the need for clean water for worship places. PROSIDING ADIWIDYA 9
SMART CITY AS AN ALTERNATIVE SOLUTION
FOR URBAN DEVELOPMENT AND EQUITY IN
INDONESIA
Talkshow Smart Education
Talkshow Smart Social Economy
Talkshow Smart Science Technology
International Webinar Smart City
ADIWIDYA 9 | #SINERGIKANPERAN
KAMIL PASCASARJANA ITB Talkshow Smart Education
Talkshow Smart Social Economy
Talkshow Smart Science Technology
International Webinar Smart City ADIWIDYA 9 | #SINERGIKANPERAN
KAMIL PASCASARJANA ITB 1. Introduction The increase in urban population has an impact on the increasing water demand and the number of public
facilities. Public facilities that must exist in one city such as health facilities, educational facilities, places of
worship, and others. The use of clean water is not only used for domestic purposes but also for non-domestic
needs or public facilities. One example of public facilities that use water to meet daily needs is a mosque. Based
on data obtained from BPS (2020) [5], Indonesia is the country with the most Muslim population in the world. As a country with a majority of Muslims, then one of the water needs in the future is the need for water to
worship. Airan Raya Mosque is one of the places of worship located in the south Lampung regency area. The mosque
was established in 2016 and has a strategic location. Over time, the infrastructure and activities around The
Airan Raya Mosque are increasing, such as the Airan Raya Mosque is close to the Sumatra Institute of
Technology College, Sukarame Police, Lampung Police, and located at the Kotabaru Toll Gate. So, The Airan
Raya Mosque can be used as a place to perform prayers both fardhu prayers and Friday prayers by the
surrounding community and travelers. This certainly has an impact on the problem of the quantity and continuity
of clean water that occurs in the Airan Raya Mosque. The water source used by the Airan Raya Mosque to fulfill daily activities comes from groundwater. Excessive use of groundwater sources by using drill wells can cause the availability of groundwater to be
drastically reduced. Exploitation of groundwater can also cause clean water difficulties because groundwater is
increasingly difficult to obtain. Rainwater harvesting system is included in one of the simple technologies, the
concept of rainwater harvesting can be used as an alternative source of clean water so as to reduce the use of
groundwater. Based on the description above, then to overcome the problem of clean water needs in the future, technology
needs to be developed about alternative water sources. One technology that can be applied is rainwater
harvesting technology. Rainwater harvesting technology is a simple technology and can be used as an effort to
reduce the use of groundwater in the Airan Raya Mosque. 1. Introduction Therefore, the author plans the application of
rainwater harvesting technology at the Airan Raya Mosque with the final result in the form of a concept and
design of rainwater harvesting system. 2.4. Calculation of Dimensions of Rainwater Harvesting System Components
In general, the components of a rainwater harvesting system consist of an capture area, a distribution system, and
a storage tank. The determination of the concept of units needed in the research of rainwater harvesting systems
in the Airan Raya Mosque is adjusted to the existing conditions of the mosque building. In this study, the
components of the system that need to be calculated dimensions are a) gutters and piping; b) first flush diverter;
c) storage tank. 2. Methods In this study, the case study used is a place of worship, namely Airan Raya Mosque located on Jalan Airan Raya,
Way Huwi Village, Jati Agung District, South Lampung Regency, Lampung. The research phase starts from
field studies, data collection, data processing and discussion, and conclusions. This research also covers the
technical aspects of the design of rainwater harvesting at the Airan Raya Mosque. The data needed in this study
include water needs data, as built drawing Masjid Airan Raya, and rain data. The rain data used in this study is
daily rainfall data from two rain stations, namely sukarame rain station and way galih rain station. 2.1. Water Demand
Calculation of clean water needs is done to find out the amount of water needed by the Airan Raya Mosque
every day. Calculation of clean water needs can be done using Equation 1. Calculation of clean water needs is done to find out the amount of water needed by the Airan Raya Mosque
every day. Calculation of clean water needs can be done using Equation 1. B = D x P . (1) Where B is water demand (liters), D is water demand per person per day (Liters/person), and P is number of
pilgrims (person) 2.2. Hydrological Analysis y
g
y
In this study hydrological analysis is needed to determine the maximum daily rainfall and intensity of rain in a
certain period of time. The stages in hydrological analysis are regional rainfall, rainfall frequency analysis,
maximum daily rainfall analysis, match testing analysis and rain intensity analysis. The results of this stage will
be used to calculate the dimensions of the components of the rainwater harvesting system at the Airan Raya
Mosque. Calculation of rain intensity can be done using equation 2. 𝐼=
𝑅24
24 (
24
𝑡)
2
3 ....... (2) 𝐼=
𝑅24
24 (
24
𝑡)
2
3 ..... (2) (2) Where I is intensity of rain (mm/hour) , R is maximum daily rainfall (mm), t is duration of rainfall (hou 2.3. Analysis of The Volume of Rainwater That Can Be Accommodated 2.3. Analysis of The Volume of Rainwater That Can Be Accommodated
Calculation of the volume of rainwater that can be accommodated is done to find out how much rainwater can be
accommodated. The amount of rainwater that can be accommodated is influenced by several factors including
rainfall that occurs in South Lampung, the area of the rain catchment, and the coefficient of runoff. S = R x A x C .......(3) (3) 2.4.3. Storage Tank In this planning to determine the volume capacity of storage tanks used a method of balance between water
supply and water demand (water balance). The main rule in determining the size of the volume of storage tanks
is that the volume of rainwater that can be accommodated must be equal to or exceed the demand for water
needs. The equation in determining the volume of the storage tank is: V = S – B (5) ....... (5) Where V is storage tank volume (m3), S is supply water (m3), B is water demand (m3) e S is supply demand (Liters), R is rainfall (mm), A is roof area (m2), C is coeffiecient runoff. Where S is supply demand (Liters), R is rainfall (mm), A is roof area (m2), C is coeffiecient runoff. 2.4. Calculation of Dimensions of Rainwater Harvesting System Components
In general, the components of a rainwater harvesting system consist of an capture area, a distribution system, and
a storage tank. The determination of the concept of units needed in the research of rainwater harvesting systems
in the Airan Raya Mosque is adjusted to the existing conditions of the mosque building. In this study, the
components of the system that need to be calculated dimensions are a) gutters and piping; b) first flush diverter;
c) storage tank. 2.4. Calculation of Dimensions of Rainwater Harvesting System Components
In general, the components of a rainwater harvesting system consist of an capture area, a distribution system, and
a storage tank. The determination of the concept of units needed in the research of rainwater harvesting systems
in the Airan Raya Mosque is adjusted to the existing conditions of the mosque building. In this study, the
components of the system that need to be calculated dimensions are a) gutters and piping; b) first flush diverter;
c) storage tank. 2.4.1. Gutter and Piping The dimensions of the gutter that will be used in the planning of the rainwater harvesting system at the Airan
Raya Mosque are guided by SNI 8153-2015 about the Plambing System in buildings. If the magnitude of the
value of the intensity of the planned rainfall is not contained in the SNI table, then to determine the area of the
planned roof can be calculated using Equation 4. Area of catchment area for planned intensity =
𝐶𝑎𝑡𝑐ℎ𝑚𝑒𝑛𝑡 𝑎𝑟𝑒𝑎 𝑖𝑛𝑡𝑒𝑛𝑠𝑖𝑡𝑦 25.4 𝑚𝑚 / ℎ𝑜𝑢𝑟
𝐷𝑒𝑠𝑖𝑟𝑒𝑑 𝑖𝑛𝑡𝑒𝑛𝑠𝑖𝑡𝑦 ( 𝑚𝑚
ℎ𝑜𝑢𝑟)/ 25,4 𝑚𝑚/ℎ𝑜𝑢𝑟 .......(4) .......(4) .(4) Area of catchment area for planned intensity =
𝐶𝑎𝑡𝑐ℎ𝑚𝑒𝑛𝑡 𝑎𝑟𝑒𝑎 𝑖𝑛𝑡𝑒𝑛𝑠𝑖𝑡𝑦 25.4 𝑚𝑚 / ℎ𝑜𝑢𝑟
𝐷𝑒𝑠𝑖𝑟𝑒𝑑 𝑖𝑛𝑡𝑒𝑛𝑠𝑖𝑡𝑦 ( 𝑚𝑚
ℎ𝑜𝑢𝑟)/ 25,4 𝑚𝑚/ℎ𝑜𝑢𝑟 2.4.2. First Flush Diverter The first flush diverter used was cylindrical of a pipe consisting of an upright pipe and a ball. In determining the
dimensions of the upright pipe used as the first flush diverter refers to SNI 8153-2005. The equation used is
equation 4 if the amount of planned rainfall intensity is not contained in the SNI table. From Table 3.1 it can be known that the total water use in one week is 26,280 liters or 26.28 m3 and the
average daily water use is 3,754 liters or 3.754 m3. 3.4.1 Gutter and Piping 3.4.1 Gutter and Piping
In determining the design of gutters and piping the value of rain intensity used is the maximum rain intensity
with a duration of rain for 2 hours and the intensity of rain with a PUH of 5 years. The result of calculating the
area of catchment for the intensity of the planned rain is 26.496 mm / hour using Equation 5. So for the roof area
of the Airan Raya Mosque of 1,050 m2 used gutters with a size of 5 inches. However, when viewed from the
availability of gutter size in the market, then in the planning of the rainwater harvesting system in the Airan Raya
Mosque used gutters with a size of 6 inches. 3.2. Hydrological Analysis Here is an example of the calculation for
December 1, 2019 : S = R x A x C
S = 6 mm x 1050 m2 x 0.85 x
1 m
1000 mm
S = 5.36 m3 S = R x A x C
S = 6 mm x 1050 m2 x 0.85 x
1 m
1000 mm
S = 5.36 m3 The same calculation was carried out for December 2, 2019 to December 31, 2019. Based on the results of
calculations, it is known that the availability of rainwater that can be accommodated in the Airan Raya Mosque
is 243.92 m3. 3.4. Design of Rainwater Harvesting System at Airan Raya Mosque 3.4.2 First Flush Diverter The first flush diverter used in the study took the form of an upright pipe consisting of an inlet and outlet. The
first flush diverter components used in this planning are fabrications available in the market. Determination of
the pipe diameter of the first flush diverter unit is done with reference to SNI 8153-2015. Thus, the size of the
diameter of the upright pipe for the first flush diverter in this study is an upright pipe with a size of 4 inches and
a length of 7 m. 3.2. Hydrological Analysis 3.2. Hydrological Analysis y
g
y
The rain data used is rain data with observation periods over the last 10 years, starting from 2011 to 2020. Based
on the distance of the rainfall observation post from the research site and the availability of data, two rain posts
were selected obtained from Balai Besar Wilayah Sungai (BBWS) Mesuji-Way Sekampung. Based on the
analysis of rainfall frequency, analysis of maximum daily rainfall, and match test, then the selected distribution
method is gumbel distribution method. Based on these data, the intensity of rain for PUH 5 years with a duration
of 2 hours is 26.496 mm / hour. y
g
y
The rain data used is rain data with observation periods over the last 10 years, starting from 2011 to 2020. Based
on the distance of the rainfall observation post from the research site and the availability of data, two rain posts
were selected obtained from Balai Besar Wilayah Sungai (BBWS) Mesuji-Way Sekampung. Based on the
analysis of rainfall frequency, analysis of maximum daily rainfall, and match test, then the selected distribution
method is gumbel distribution method. Based on these data, the intensity of rain for PUH 5 years with a duration
of 2 hours is 26.496 mm / hour. Table 3.2 Maximum rainfall plan
Year
Maximum Rainfall Plan (mm)
Year
Maximum Rainfall Plan (mm)
2011
38
2016
61.7
2012
42
2017
81.5
2013
143.5
2018
56.5
2014
36.5
2019
155
2015
56.6
2020
87.5 Table 3.2 Maximum rainfall plan 3.3. Analysis of The Volume of RainWater That Can Be Accommodated 3.3. Analysis of The Volume of RainWater That Can Be Accommodated
In this study, the volume of rainwater that can be accommodated to find out how much rainwater availability to
meet the needs of clean water in the Airan Raya Mosque. The amount of the value of water availability or the
volume of rainwater that can be accommodated is influenced by the amount of rainfall that occurs in Lampung,
the area of the building roof as a rain catchment area, and the coefficient of runoff. The rain data used is the
maximum rain data that occurred in 2019 to be exact in December. 3.1. Water Demand The basic planning used in accounting for the volume of water demand is to use the method of number of users. According to RSNI T-01-2003) each pilgrim spends about 15 liters of water per day. Based on the results of field
surveys for one week, the calculation of water needs in the Airan Raya Mosque in one day can be done by
multiplying the number of worshipers in one day by the standard of water use per day through equation 1. Here
are the water needs at the Airan Raya Mosque on Monday : B = D x P
B = 15 liters/person/day x 183 person
B = 2,745 liters/day
B = 2.745 m3/day B = D x P
B = 15 liters/person/day x 183 person
B = 2,745 liters/day
B = 2.745 m3/day Table 3.1 Recapitulation of daily water use
Days
Number Of Pilgrims
(person)
Standard Daily Water Usage
(Liters)
Water Use
(Liters)
Monday
183
15
2745
Tuesday
193
15
2895
Wednesday
178
15
2670
Thursday
178
15
2670
Friday
587
15
8805
Saturday
200
15
3000
Sunday
233
15
3495
Sum
26280
Average
3754 Table 3.1 Recapitulation of daily water use From Table 3.1 it can be known that the total water use in one week is 26,280 liters or 26.28 m3 and the
average daily water use is 3,754 liters or 3.754 m3. 3.4.3 Storage Tank Storage tanks are the main component in the planning of rainwater harvesting systems. This is because storage
tank components require the most cost compared to other components. Therefore, it is very important to plan the
appropriate storage tank capacity in order to save costs. Determination of the dimensions or volume of rainwater storage tanks in this planning is done using the method of balance between water supply and water demand
(water balance) in The Airan Raya Mosque. Here is an example of a calculation for December 1, 2019 : storage tanks in this planning is done using the method of balance between water supply and water demand
(water balance) in The Airan Raya Mosque. Here is an example of a calculation for December 1, 2019 : V = S – B
V = 5.36 m3 – 3.754 m3
V = 1,61 m3 V = S – B V = 5.36 m3 – 3.754 m3 V = 5.36 m3 – 3.754 m3
V = 1,61 m3 Furthermore, it was done in the same way for December 2, 2019 to December 31, 2019. So, in the planning of
the rainwater harvesting system at the Airan Raya Mosque was obtained the required storage tank volume size of
3.15 m3 / day. In the planning of the rainwater harvesting system at the Masjid Airan Raya the storage tank used
is a storage tank with a size of 3,100 liters. Picture 3.1 Front view of the rainwater harvesting system at the Airan Raya mosque Picture 3.1 Front view of the rainwater harvesting system at the Airan Raya mosque 4. Conclusion Based on the results of calculations and planning, in this study, the volume of rainwater that can be harvested by
the roof of the Airan Raya Mosque based on the calculation results is 243.92 m3 per month. As for the water
demand at the Airan Raya Mosque is 3,754 liters/day, the specifications of the planned rainwater harvesting
system are a storage tank with a size of 3,100 liters, a rectangular gutter with a size of 6 inches, and a first flush
diverter with a size of 4 inches and 7m long. 3.5. Maintenance of Rainwater Harvesting System 3.6. The process of maintaining and monitoring the rainwater harvesting system needs to be done
periodically so that the rainwater harvesting system can be implemented sustainably and ensure that
no damage can interfere with the running of the system. The treatment and monitoring process that
needs to be done can be seen in the Table. 3.3 Table 3.3 Maintenance Of Rainwater Harvesting System
No
Component
Tipe Of Maintanance
Period
1
Roof
Ensure the roof is in good condition and does
not leak or rust. 6 month
2
Gutter
Ensure the condition of the gutter is good and
not damaged. In addition, make sure the gutter
condition is no blockage and the entire gutter
connection is not loose. 6 month
3
First Flush Diverter
Ensure the unit is not damaged and does not
leak
6 month No
Component
Tipe Of Maintanance
Period
4
Storage Tank
Ensure the tank doesn't leak and crack
6 month
Drain the tank periodically
1 x 2 month and at the
beginning of the rainy season
or the transition of the
seasons
5
Filtration
Wash or replace the filter media
1 x 1 month or according to
needs References [1] Pala, G.K., Pathivada, A.P., Velugoti, S.J.H., Yerramsetti, C., and Veeranki, S., “Rainwater Harvesting – A
Review On Conservation, Creation & Cost Effectiveness”, (Material Today: Proceedings, Elsevier, 2021),
vol 45, pp. 6567 – 6571. [2] Crosson, C., Tong, D., Zhang, Y., and Zhong, Q., “Rainwater as A Renewable Resource To Achieve Net
Zero Urban Water In Water Stressed Cities”, (Resources, Conservation &Recycling, Elsevier, 2021), vol
164, 105203. [3] Azmi, N.A., Kandar, M.Z. “Factors contributing in the design of environmentally sustainable mosques”,
(Elsevier, Journal of Building Engineering, 2019) vol 23, pp. 23-37. [4] Campisano, A., Butler, D., Ward, S., Burns, M.J., Friedler, E., DeBusk, K., Jeffes, L.N.F., Ghisi, E.,
Rahman, A., Furumai, H., Han, M., “Urban Rainwater Harvesting Systems: Research, Implementation
And Future Perspectives”, (Elsevier, Water Research, 2017), vol 115, pp. 195-209. [5] Badan Pusat Statistik, “Sensus Penduduk Indonesia Tahun 2020”, 21 Januari 2021, [Online],
Available at : https://www.bps.go.id/ [6] Gatot Eko Susilo. “Rainwater Harvesting as Alternative Source for Wudlu Water in Indonesia”
(Civil and Enviromental Science Journal, 2018), vol.1, no.2, pp. 62-69. [7] Badan Standardisasi Nasional, “RSNI T-01-2003 Tata Cara Perencanaan Plumbing, Jakarta: Standa
Nasional Indonesia, 2003. [8] Suripin, “Sistem Drainase Perkotaan yang Berkelanjutan”, Yogyakarta: ANDI Yogyakarta, 2004. [9] Menteri Negara Lingkungan Hidup, “PerMenLH Nomor 12 Tahun 2009 Tentang Pemanfa
Hujan”, 15 April 2009. [10] Imroatul Chalimah Juliana, Ari Taufik Gunawan dan Sarino, “Dasar-Dasar Penerapan Sistem
Rainwater Harvesting (RWH)”, Palembang: Universitas Sriwijaya, 2019. [11] Agus Maryono, “Memanen Air Hujan (Rainwater Harvesting)”, Yogyakarta: Gadjah Mada University
Press, 2016. [12] Nazharia, C. dan Sri, M., “Perhitungan Pembiayaan Pemanenan Air Hujan sebagai Sistem Penyediaan
Air Bersih dalam Berbagai Skala di Kelurahan Sukajadi Kota Dumai’, (Jurnal Perencanaan Wilayah dan
Kota B SAPPK, 2014), vol 2, pp. 79-88. [13] Badan Standardisasi Nasional, “SNI 8153-2015: Sistem Plambing Pada Bangunan Gedung”, Jakarta:
Standar Nasional Indonesia, 2015. [14] Sosrodarsono Suyono dan Kensaku Takeda “Hidrologi untuk Pengairan”, Pradnya Pramita
Jakarta, 2003. [15] Sunjoto, “Sistem Pengelolaan Limbah Air Hujan”, Yogyakarta: Universitas Gadjah Mada, 1988. [16] Fitri, Wulandari, “Perancangan Sistem Pemanenan Air Hujan di Kampus ITB Jatinangor”,
Bandung: Institut Teknologi Bandung, 2017. Prosiding Seminar Nasional Adiwidya 8 Pascasarjana ITB
1
7 November 2020
Alamat Redaksi
Keluarga Mahasiswa Islam (KAMIL) Pascasarjana ITB,
Gedung Kayu Lt. 2, Kompleks Masjid Salman ITB,
Jl. Ganesha No. 10, Bandung,
40132 https://kamilpasca.itb.ac.id/ Alamat Redaksi Alamat Redaksi
Keluarga Mahasiswa Islam (KAMIL) Pascasarjana ITB,
Gedung Kayu Lt. 2, Kompleks Masjid Salman ITB,
Jl. Ganesha No. 10, Bandung,
40132 https://kamilpasca.itb.ac.id/ 1
|
https://openalex.org/W2740274371
|
http://eprints.whiterose.ac.uk/121373/1/Spence_et_al-2017-Ecology_and_Evolution.pdf
|
English
| null |
Making the most of survey data: Incorporating age uncertainty when fitting growth parameters
|
Ecology and evolution
| 2,017
|
cc-by
| 9,195
|
K E Y W O R D S Bayesian statistics, Clupea harengus, fisheries stock assessment, Gadus morhua, growth, seasonal
growth, survey data, uncertainty analysis, von Bertalanffy growth function In many areas, growth can be described using the von Bertalanffy
growth function (VBGF) (von Bertalanffy, 1957). It describes an ani-
mal’s size, li, as a function of its age, ai, Received: 17 January 2017 | Revised: 31 May 2017 | Accepted: 3 July 2017 Received: 17 January 2017 | Revised: 31 May 2017 | Accepted: 3 July 2017 Received: 17 January 2017 | Revised: 31 May 2017 | Accepted: 3 July 2017
DOI: 10.1002/ece3.3280 DOI: 10.1002/ece3.3280 Making the most of survey data: Incorporating age uncertainty
when fitting growth parameters Michael A. Spence1,2
| Alan J. Turtle1 Abstract
Individual growth is an important parameter and is linked to a number of other biologi-
cal processes. It is commonly modeled using the von Bertalanffy growth function
(VBGF), which is regularly fitted to age data where the ages of the animals are not
known exactly but are binned into yearly age groups, such as fish survey data. Current
methods of fitting the VBGF to these data treat all the binned ages as the actual ages. We present a new VBGF model that combines data from multiple surveys and allows
the actual age of an animal to be inferred. By fitting to survey data for Atlantic herring
(Clupea harengus) and Atlantic cod (Gadus morhua), we compare our model with two
other ways of combining data from multiple surveys but where the ages are as re-
ported in the survey data. We use the fitted parameters as inputs into a yield-per-
recruit model to see what would happen to advice given to management. We found
that each of the ways of combining the data leads to different parameter estimates for
the VBGF and advice for policymakers. Our model fitted to the data better than either
of the other models and also reduced the uncertainty in the parameter estimates and
models used to inform management. Our model is a robust way of fitting the VBGF
and can be used to combine data from multiple sources. The model is general enough
to fit other growth curves for any taxon when the age of individuals is binned into
groups. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium,
provided the original work is properly cited. reaches this value, and t0 is the theoretical age (in years) when the an-
imal was length 0 (Beverton & Holt, 1959; Schnute & Fournier, 1980). seasons and years. However, parameter values can be sensitive to how
the data from different surveys are combined (Wilson et al., 2015). Figure 1 shows equation (2) fitted to data from the Scottish West
Coast Surveys International Bottom Trawl Surveys (SWC-IBTS) for
Atlantic herring (Clupea harengus) (see Section 2 for more information)
collected in quarter 1 only, quarter 4 only, and both quarters combined
(ICES, 2016b). Combining the data from both surveys changes the fit
of the VBGF considerably with a much lower growth rate and much
higher asymptotic length. This suggests that simply combining survey
data from the two survey times can lead to fitting statistically inconsis-
tent models, and thus, a more appropriate method of combining them
needs to be explored. seasons and years. However, parameter values can be sensitive to how
the data from different surveys are combined (Wilson et al., 2015). Figure 1 shows equation (2) fitted to data from the Scottish West
Coast Surveys International Bottom Trawl Surveys (SWC-IBTS) for
Atlantic herring (Clupea harengus) (see Section 2 for more information)
collected in quarter 1 only, quarter 4 only, and both quarters combined
(ICES, 2016b). Combining the data from both surveys changes the fit
of the VBGF considerably with a much lower growth rate and much
higher asymptotic length. This suggests that simply combining survey
data from the two survey times can lead to fitting statistically inconsis-
tent models, and thus, a more appropriate method of combining them
needs to be explored. g
(
,
;
,
)
The VBGF is used extensively to describe fish species (Chen,
Jackson, & Harvey, 1992; Essington, Kitchell, & Walters, 2001). The
parameters, often fitted to survey data, can be used as inputs to
single species and multispecies models used to inform management
and policymakers (Blanchard et al., 2014; Pardo et al., 2013; Thorpe,
Le Quesne, Luxford, Collie, & Jennings, 2015). When deciding on a
new policy, the decision maker needs to know what all the likely
consequences of that policy are, therefore, it is essential to quantify
uncertainty in these models (Harwood & Stokes, 2003). As uncer-
tainty in the VBGF parameters can filter through to uncertainty in
the models used by the management, it is important to quantify this
robustly (Pardo et al., 2013; Walters & Martell, 2004). Siegfried and
Sansó (2006) and Hamel (2015) presented a probabilistic version of
the VBGF, An alternative way of combining data is to increase the age of
fish according to the time of year at which the fish was surveyed
(Chambers, Sidhu, O’Neil, & Sibanda, 2017; Sparre & Venema, 1998). In effect, this assumes that every fish spawned on January 1st, and its
age is the number of winters survived plus the time of year at which it
was caught, expressed as proportion. (2)
li =l∞(1−exp{−k(ai −t0)})ϵ, (2) where ϵ is a multiplicative error and takes account of all the uncer-
tainty not explained by the VBGF (e.g., individual growth and tempera-
ture effects) as well as measurement errors in the data with In the survey data, the age of a fish and its length are recorded. The
age of an individual is estimated by counting the number of layers of
tree ring-like growth in the otolith (ICES, 2016b), which is essentially
the number of winters it has survived (Pardo et al., 2013). However,
in the VBGF, age is a continuous variable but in the survey data, fish
are usually binned in yearly groups (see the data points in Figure 1). A
fish having survived t winters caught qth through the year is aged in
the region [t−1+q,t+q]. This means that two fish binned in the same
yearly group, could differ by up to a year. For most species, fish that log ϵ∼N(0,σ2), log ϵ∼N(0,σ2), and fitted it using a Bayesian framework enabling the uncertainty
to be quantified. and fitted it using a Bayesian framework enabling the uncertainty
to be quantified. As increased certainty can lead to “better” decisions, it is, therefore,
desirable to fit the VBGF to as much of the survey data as possible. This may involve combining data from different surveys over different FIGURE 1 The top plot shows the data
and the line with the maximum posterior
density sampled from the Markov Chain
Monte Carlo (MCMC) when fitted to data
from quarter 1 only, quarter 4 only, and all
of the data. 1 | INTRODUCTION Throughout ecology, growth is an important parameter that describes
the life history of individuals and species (Austin, Robinson, Robinson,
& Ricklefs, 2011; Einum, Forseth, & Finstad, 2012; Paine et al., 2012;
Pardo, Cooper, & Dulvy, 2013). It is often linked to natural mortality
(e.g., Gislason, Daan, Rice, & Pope, 2010; Pauly, 1980), size-based sur-
vival (e.g., Lorenzen, 2000), life span (e.g., Hoenig, 1983), and expected
abundances (e.g., Andersen & Beyer, 2006). (1)
li =l∞(1−exp{−k(ai −t0)}). (1) Here, l∞ is the asymptotic size of the animal (in millimeters), the ex-
pected size that an individual will reach if it was to live forever, k, the
growth coefficient (per year), describes the rate in which the individual This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium,
provided the original work is properly cited. he terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium,
cited. Ecology and Evolution. 2017;7:7058–7068. SPENCE and TURTLE 7059 The bottom four plots show
the marginal posteriors for each of the four
parameters for each of the three MCMC
runs
10
0
2
4
6
8
12
14
50
100
200
300
400
Age (years)
Length (mm)
Q1 only
Q4 only
All data
0.3
0.4
0.5
0.6
0.7
0.8
0
50
100
150
k
Density
290
300
310
320
330
0.0
0.2
0.4
0.6
0.8
l∞
Density
−1.5
−1.0
−0.5
0.0
50
0
10 20 30 40
t0
Density
0.008 0.010 0.012 0.014 0.016 0.018 0.020
0
1000
3000
σ2
Density 10
0
2
4
6
8
12
14
50
100
200
300
400
Age (years)
Length (mm)
Q1 only
Q4 only
All data
0.3
0.4
0.5
0.6
0.7
0.8
0
50
100
150
k
Density
290
300
310
320
330
0.0
0.2
0.4
0.6
0.8
l∞
Density
−1.5
−1.0
−0.5
0.0
50
0
10 20 30 40
t0
Density
0.008 0.010 0.012 0.014 0.016 0.018 0.020
0
1000
3000
σ2
Density FIGURE 1 The top plot shows the data
and the line with the maximum posterior
density sampled from the Markov Chain
Monte Carlo (MCMC) when fitted to data
from quarter 1 only, quarter 4 only, and all
of the data. The bottom four plots show
the marginal posteriors for each of the four
parameters for each of the three MCMC
runs σ2 7060 SPENCE and TURTLE We define ti to be the number of winters that individual i has sur-
vived before being surveyed qith of the way through the year, with
qi ∈[0,1), and li to be the length that the individual was when it was
surveyed. have survived the same number of winters are not necessarily the
same age due to differences in spawning times, for which there has
been empirical studies conducted (e.g., Brander, 1994; Knijn, Boon,
Heessen, & Hislop, 1993). Assuming that all fish in the same yearly group are exactly the
same age would mean that we are overly confident about the age of
the fish. This could have a large effect on estimates of the parameters
when fitting the VBGF to this kind of data. Cope & Punt (2007) and
Doll, Lauer, & Clark-Kolaks (2017) included Gaussian random effect
terms that were used to account for uncertainty in the age of individu-
als when fitting the VBGF. 2.2 An individual spawned at time si, hav-
ing survived ti winters and being surveyed in qi would be effectively
aged a
i =ti −f(si)+f(qi) for the purpose of calculating growth from
the VBGF. Management advice can be sensitive to the parameters of the
VBGF. In order to demonstrate this, we used the parameter estimates
as inputs to a yield-per-recruit (YPR) model (Gabriel, Sissenwine,
& Overholtz, 1989) often used to advise management (Doll et al.,
2017). In Section 2, we introduce the spawning and seasonal growth ver-
sions of the VBGF and describe a simulation study that demonstrates
the effects of ignoring uncertainty in the ages when fitting the VBGF. We also compare how the new models perform compared to other
VBGF that combine data from different surveys by fitting to herring
and cod data. The results of the simulation study as well as the pos-
terior distributions of the different versions of the VBGF and their
effects on the YPR analysis are presented in Section 3. We conclude
with a discussion in Section 4. For example, Somers (1988)’s version of the seasonal VBGF, li =l∞(1−exp{−k(ai −t0)−S(ai)+S(t0)}), with S(ai)= ck
2π sin (2π(ai −ts)), has has f(x)=x+ c
2π sin (2π(x−ts)) 2.1 We define si to be the spawning time of the ith fish. Datta & Blanchard
(2016) said that the spawning times of an individual from a single spe-
cies are (3)
si ∼vonMises(μ,τ), (3) where si ∈[0,1). The von Mises distribution is a continuous circular
distribution with location parameter μ and scale parameter τ on the
space (0,1) (Best and Fisher, 1979). Therefore, a fish surveyed in qi
having survived ti winters was aged ti −si +qi when it was surveyed. It is intuitive to think that of two fish, aged
the same in the survey data but of different lengths, the larger fish
spawned earlier than the smaller, especially if they were young. With
this in mind, in this study, we develop a model that uses information
from previous studies about the spawning times of species (Datta &
Blanchard, 2016) to infer the age of individual fish and allow multiple
surveys to be combined. 2.2 Growth does not necessarily follow the VBGF constantly but may
be seasonal (García-Berthou et al., 2012). If we assume that from
year to year an individual fish follows the VBGF, that is one year
from the current time, an individual’s growth is described by the
VBGF, but is not throughout the year. Let x be the period through
the year and f(x) be the proportion of von Bertalanffy growth that
has already occurred during that year. This means that f(x)∈[0,1]
and x∈[0,1) with f(0)=0, as at the start of the year an individual has
not grown yet, and f(1)=1 as at the end of the year the individual
has completed its annual growth. With this in mind, we re-write
equation (2) as By fitting our model and two other alternative versions of VBGF, to
SWC-IBTS data for Atlantic herring (Clupea harengus), a species with a
long-spawning season, and Atlantic cod (Gadus morhua), a species with
a short-spawning season (Datta & Blanchard, 2016), we demonstrate
that the parameters can be sensitive to the version of the VBGF that
we fit them to. (4)
li =l∞(1−exp{−k(a
i −t
0)})ϵ, (4) where a
i =⌊ai⌋+f(ai −⌊ai⌋) and t
0 =⌊t0⌋+f(t0 −⌊t0⌋). ⌊y⌋ is the floor
function of y. Notice that ai is replaced with a′
i, and t0 is replaced
with t′
0, as a′
i and t′
0 are the theoretical ages at which an individual
will be if it followed the von Bertalanffy growth curve at age ai and
t0, respectively. We have defined f(1)=1, therefore the interpreta-
tion of k will remain the same. An individual spawned at time si, hav-
ing survived ti winters and being surveyed in qi would be effectively
aged a
i =ti −f(si)+f(qi) for the purpose of calculating growth from
the VBGF. where a
i =⌊ai⌋+f(ai −⌊ai⌋) and t
0 =⌊t0⌋+f(t0 −⌊t0⌋). ⌊y⌋ is the floor
function of y. Notice that ai is replaced with a′
i, and t0 is replaced
with t′
0, as a′
i and t′
0 are the theoretical ages at which an individual
will be if it followed the von Bertalanffy growth curve at age ai and
t0, respectively. We have defined f(1)=1, therefore the interpreta-
tion of k will remain the same. 2.2 | Seasonal growth curve Growth may not follow the VBGF but can be seasonal (García-
Berthou, Carmona-Catot, Merciai, & Ogle, 2012). By making ai in
equation (1), the theoretical age that the individual would be if they
grew according to the VBGF, as opposed to their actual age, we can
fit the VBGF with seasonal growth. Furthermore, constraining the dif-
ference of a year in theoretical age and actual age to be the same, the
interpretation of k would remain unchanged. This involves describing
how growth occurs throughout a year and is a generalization of many
other seasonal variations of VBGF (e.g., Pauly, Soriano-Bartz, Moreau,
& Jarre-Teichmann, 1992; Somers, 1988). 1989; Nelson, 2016) in R (R Core Team, 2015). The length at age a in
the YPR model was the expected length between age a and a+1, that is, In this study, we take f to have a more flexible form, a linear inter-
polator of m+1 points, corresponding to the number of surveys per-
formed in a single year, m, la =l∞exp
(
σ2
2
) (
1−1
k exp (−kt0)( exp (−ka)−exp (−k(a+1)))
)
, (5)
f(x)=
( cj −cj−1
qj −qj−1
)
(x−qj)+cj, (5) where k, l∞, t0, and σ2 were sampled from the posterior distribu-
tion. A description of the YPR model can be found in the Supporting
Information. As we were only interested in the sensitivity of the von
Bertalanffy parameters on the YPR model we fixed the other inputs. The length weight ratios were taken from FishBase (Froese & Pauly,
2016), and the mortality rates were taken from ICES (2016a,b) for her-
ring and cod, respectively. Table 1 gives a summary of the parameter
values we used when fitting the models. for j=1 … m with qj−1 ≤x<qj, x1 =c1 =0 and qm =cm =1. for j=1 … m with qj−1 ≤x<qj, x1 =c1 =0 and qm =cm =1. 2.3 | Data The SWC-IBTS data for Atlantic herring (Clupea harengus) and Atlantic
cod (Gadus morhua) from 2001 to 2014 were extracted from DATRAS
(2016), with surveys performed in quarter 1 (qi =0) and for quarter 4
(qi =0.75). 2.4 | Simulation study We fitted the VBGF with ai =ti for quarter 1 only and quarter 4 only
data as well as all of the data combined for herring. In order to demonstrate the effect of ignoring the age uncertainty
we fitted the VBGF to simulated data. Assuming that the mortality
rate of fish is unchanged regardless of when an individual spawned,
the marginal distribution of the ages of the fish will then be ti −si +qi. We sampled ti and qi from age distributions found in the SWC-IBTS
data and si from equation (3). Each individual’s growth was assumed
to follow the VBGF with its length sampled from equation (2) with
ai =ti −si +qi, k=0.60, l∞=293.3, t0 =−0.713, σ2 =0.0572, μ=0.868,
and τ=0.404 for herring and with k=0.24, l∞=1143, t0 =−0.179,
σ2 =0.1392, μ=0.312, and τ=5.473 for cod. We then fitted equa-
tion (2) with ai =ti and ai =ti +qi for 102, 103, 104, 105, and 106
randomly selected fish. We compared the different variations of combining data when
fitting the VBGF. As well as fitting equation (2) with ai =ti −si +qi, as
described in Section 2.1, we fitted the equation with ai =ti, that is the
age of the fish is the number of winters that it has survived, and with
ai =ti +qi (Sparre & Venema, 1998). We also fitted the data to equa-
tion (4) with a
i =ti +f(qi) and a
i =ti −f(si)+f(qi). In summary, we fitted
the following models to the herring and cod data: I Equation (2) with ai =ti
II Equation (2) with ai =ti +qi
III Equation (4) with a
i =ti +f(qi)
IV Equation (2) with ai =ti −si +qi
V Equation (4) with a
i =ti −f(si)+f(qi) I Equation (2) with ai =ti
II Equation (2) with ai =ti +qi
III Equation (4) with a
i =ti +f(qi)
IV Equation (2) with ai =ti −si +qi
V Equation (4) with a
i =ti −f(si)+f(qi) I Equation (2) with ai =ti
II Equation (2) with ai =ti +qi
III Equation (4) with a
i =ti +f(qi)
IV Equation (2) with ai =ti −si +qi
V Equation (4) with a
i =ti −f(si)+f(qi) 2 | METHODS and In this section, we introduce the models that we are going to use to
compare when fitting to simulated and survey data from SWC-IBTS. t
0 =t0 −c
2π sin (2π(t0 −ts)). SPENCE and TURTLE 7061 σ2 ∼inv- gamma(0.001, 0.001). In models III and V, c2 ∈[0,1] uniformly. A full description of
the prior and likelihood for each of the models can be found in the
Supporting Information (Section A). We compared the fitted models using the Watanabe-Akaike infor-
mation criterion (WAIC) (Watanabe, 2010) defined as Models II and III gave roughly the same values of YPR. Model I re-
duces the YPR by 11% and models IV and V increased the YPR a large
amount. The standard deviation for models IV and V is large as the YPR
is actually bimodal. −2
(
−
N
∑
i=1
log (E(p(li|훉)))+2
N
∑
i=1
E( log (p(li|훉)))
)
, where θ is the parameters of the model, and the expectation is taken
over the posterior distribution. The line in Figure 3 shows the VBGF fitted with the parameters
from the posterior mean, and the points show the mean age of the
fish when model IV was fitted. It also shows the posterior predictive
distribution. The plot shows that length gives more information about
an individuals age earlier in its life. As the posterior distribution cannot be derived analytically, we
are required to sample from it using a Markov Chain Monte Carlo
(MCMC) algorithm. Due to correlations in the posterior distribu-
tion, we used the No-U-turn Hamiltonian Monte Carlo algorithm
(Gelman, Lee, & Guo, 2015; Hoffman & Gelman, 2014). After a
burn-in period of 1,000 iterations, we sampled 1,000 points from
the posterior distribution. Visual examination through trace and au-
tocorrelation plots shows that for each of the models the MCMC
sampler reached its stationary distribution. Furthermore, when four
chains were run for each model, ̂R, Gelman’s indexes (Gelman &
Rubin, 1992), were close to 1 for all of the parameters in all of the
models indicating that the samplers were mixing well and sampling
from the same distribution. Figure 4 shows the length residuals of the posterior mean of model
V fitted to herring data and simulated data. The two plots appear to
be similar suggesting that the assumption of log-normal multiplicative
errors seems reasonable. 3.3 | Cod Table 3 shows summaries of the posterior distribution for cod for
each of the different models, the YPR relative to model II and the
WAIC. The posterior means of c2 were 0.78 and 0.81 with standard
deviations 0.02 for models III and V, respectively. For model I, k was
much lower than the other versions, and l∞ is much larger. Models IV
and V gave higher values and more uncertainty about k than the other
models, but gave more certainty on the other parameters. They also
fit the data better than any other method of fitting the combined data,
as shown by the WAIC. to quarter 1 data and quarter 4 data only resulted in different posterior
distributions. Fitting to the whole dataset did not yield a combination
of the two posterior distributions but actually gives a very different
posterior distribution. Figure 1 demonstrates that there needs to be a
more coherent way of combining data from different surveys. For all of the models we put weak prior information on the pa-
rameters. We used a uniform prior for k such that k∈[0,3], for l∞ and
t0 we used improper priors such that they could equally take any
value in [0,∞) and (−∞,0], respectively. The prior for the variance
term was Table 2 shows the summaries of the posterior distributions, the
YPR for Fmax, the fishing level that lead to the maximum yield, relative
to model II and the WAIC for all five models fitted to the herring data
described in Section 2.6. The posterior means of c2 were 0.91 and 0.63
with standard deviations 0.005 and 0.004 for models III and V, respec-
tively. Models IV and V have lower uncertainty on the parameters than
any of the other methods as well as fitting the data considerably better
as shown by the WAIC. 3.1 | Simulation study The marginal posterior distributions of the parameters of model II
fitted to increasing amounts of simulated herring data are shown in
Figure 2 with the solid line showing truth. As the amount of data in-
creases, we get more certain about the wrong value. This suggests
that ignoring the uncertainty in the ages leads to the model not
being consistent. We found a similar result when fitting model II to
the simulated cod data. Similarly we found model I to be inconsistent
when fitted to both herring and cod simulated data (see Supporting
Information). Figure 5 shows the marginal posterior distributions of the spawn-
ing times of two cod, with ti =1 and qi =1, fitted to model IV. The two
fish are of different lengths, and the posterior probability that the
larger fish spawned before the smaller is 0.99 with the mean differ-
ence in their ages being 0.26 years. 2.5 | Yield-per-recruit The parameters of the von Mises distributions in models IV and V
were taken from Datta & Blanchard (2016). In models III and V, f(⋅) was
defined in equation (5), with m=3, x2 =0.75, and f(0.75)=c2. In order to investigate how sensitive management advice can be to
the choice of VBGF model, we used the fitted parameter values as
inputs for a YPR analysis. This was conducted following the modified
Thompson-Bell algorithm using the fishmethods package (Gabriel et al., TABLE 1 The input parameters for the
models. The length weight conversion is
given by w=αlβ Parameter
Herring
Cod
Meaning
N
22,790
2,815
Number of individuals surveyed
q1
18,933
1,910
Number of individuals surveyed in quarter 1
q4
3,857
905
Number of individuals surveyed in quarter 4
μ
0.858
0.312
Location parameter of spawning time
τ
0.405
5.473
Scale parameter of spawning time
α
0.006
0.008
Length–weight parameter
β
3.05
3.06
Length–weight parameter
M0
0.767
0.537
Mortality aged 0
M1
0.385
0.386
Mortality aged 1
M2
0.356
0.306
Mortality aged 2
M3
0.339
0.262
Mortality aged 3
M4
0.319
0.237
Mortality aged 4
M5
0.314
0.223
Mortality aged 5
M6+
0.307
0.211
Mortality aged 6+ 7062 SPENCE and TURTLE 3.2 | Herring The solid
line shows the true value of the parameters
k
0.50
0.55
0.60
0.65
102
103
104
105
106
102
103
104
105
106
l∞
285
290
295
300
305
t0
−0.6
−0.4
−0.2
0.0
σ
0.00
0.02
0.04
0.06
Sample size
102
103
104
105
106
102
103
104
105
106
Model
I
II
III
IV
V
k
Mean
0.30
0.58
0.62
0.60
0.60
SD
0.003
0.003
0.004
0.003
0.003
l∞
Mean
323.2
294.3
291.9
293.3
293.3
SD
1.02
0.48
0.42
0.27
0.27
t0
Mean
−1.396
−0.179
−0.072
−0.713
−0.672
SD
0.014
0.007
0.007
0.006
0.006
σ
Mean
0.137
0.098
0.096
0.057
0.056
SD
0.00067
0.00047
0.00047
0.00039
0.00038
% increase YPR
Mean
−11
0
1
23
22
SD
0.4
0
0.4
0.9
0.9
WAIC
−25,930
−41,119
−42,244
−62,582
−62,966
VBGF, von Bertalanffy growth function; WAIC, Watanabe-Akaike information criterion; YPR,
yield-per-recruit. TABLE 2 The posterior mean and
standard deviation of the parameters of
the VBGF, the percentage increase in YPR
with fishing at Fmax, relative to model II,
and the WAIC for the different models
fitted to herring data | 7063
k
0.50
0.55
0.60
0.65
102
103
104
105
106
102
103
104
105
106
l∞
285
290
295
300
305
t0
−0.6
−0.4
−0.2
0.0
σ
0.00
0.02
0.04
0.06
Sample size
102
103
104
105
106
102
103
104
105
106 SPENCE and TURTLE 7063 | 7063
l∞
285
290
295
300
305
102
103
104
105
106 | 7063
SPENCE and TURTLE
FIGURE 2 The von Bertalanffy
growth function fitted to simulated data
with ai =ti +qi. The violin plots (Hintze &
Nelson, 1998) show the marginal posterior
distributions for each of the parameters
fitted to different sample sizes. The solid
line shows the true value of the parameters
k
0.50
0.55
0.60
0.65
102
103
104
105
106
102
103
104
105
106
l∞
285
290
295
300
305
t0
−0.6
−0.4
−0.2
0.0
σ
0.00
0.02
0.04
0.06
Sample size
102
103
104
105
106
102
103
104
105
106 k
0.50
0.55
0.60
0.65
102
103
104
105
106 102
103
104
105
106
t0
−0.6
−0.4
−0.2
0
σ
0.00
0.02
0.04
0.06
Sample size
102
103
104
105
106 FIGURE 2 The von Bertalanffy
growth function fitted to simulated data
with ai =ti +qi. 3.2 | Herring The violin plots (Hintze &
Nelson, 1998) show the marginal posterior
distributions for each of the parameters
fitted to different sample sizes. The solid
line shows the true value of the parameters Model
I
II
III
IV
V
k
Mean
0.30
0.58
0.62
0.60
0.60
SD
0.003
0.003
0.004
0.003
0.003
l∞
Mean
323.2
294.3
291.9
293.3
293.3
SD
1.02
0.48
0.42
0.27
0.27
t0
Mean
−1.396
−0.179
−0.072
−0.713
−0.672
SD
0.014
0.007
0.007
0.006
0.006
σ
Mean
0.137
0.098
0.096
0.057
0.056
SD
0.00067
0.00047
0.00047
0.00039
0.00038
% increase YPR
Mean
−11
0
1
23
22
SD
0.4
0
0.4
0.9
0.9
WAIC
−25,930
−41,119
−42,244
−62,582
−62,966
VBGF, von Bertalanffy growth function; WAIC, Watanabe-Akaike information criterion; YPR,
yield-per-recruit. TABLE 2 The posterior mean and
standard deviation of the parameters of
the VBGF, the percentage increase in YPR
with fishing at Fmax, relative to model II,
and the WAIC for the different models
fitted to herring data TABLE 2 The posterior mean and
standard deviation of the parameters of
the VBGF, the percentage increase in YPR
with fishing at Fmax, relative to model II,
and the WAIC for the different models
fitted to herring data TABLE 2 The posterior mean and
standard deviation of the parameters of
the VBGF, the percentage increase in YPR
with fishing at Fmax, relative to model II,
and the WAIC for the different models
fitted to herring data VBGF, von Bertalanffy growth function; WAIC, Watanabe-Akaike information criterion; YPR,
yield-per-recruit. the spawning season, but are recorded as being the same age in
the survey data. We presented a model that includes information
regarding the spawning period and demonstrated that our model
improved the fit of the VBGF for herring and cod off the West Coast
of Scotland. values of σ for herring, where we fitted the VBGF to much more data,
than for cod. This increase in certainty carries through to the YPR
model and is demonstrated by the dramatic increase in certainty from
the herring YPR model compared to cod. In the survey data, the age of the fish is recorded as the
number of winters that it has survived. Therefore, fish caught in
quarter 1 and quarter 4 that have survived the same number of
winters are recorded as being the same age. 3.2 | Herring In this study, we introduced a new model that uses information
about spawning times to improve the fit of the VBGF. We demon-
strated that fitting to multiple surveys is a good way of reducing
uncertainty when fitting the VBGF but existing methods of doing
this are unsatisfactory. Difficulties are caused because fish that are
caught at the same time can be different ages, due to variations in Figure 1 shows the growth curve corresponding to the posterior mode
and the marginal posterior distributions of the four parameters when
equation (2) was fitted to the length and age data of herring for quar-
ter 1 only, quarter 4 only, and both combined. The bottom 4 plots
show the posterior density for the parameters of equation (2). Fitting | 7063
SPENCE and TURTLE
the spawning season, but are recorded as being the same age in
the survey data. We presented a model that includes information
regarding the spawning period and demonstrated that our model
improved the fit of the VBGF for herring and cod off the West Coast
of Scotland
values of σ for herring, where we fitted the VBGF to much more data,
than for cod. This increase in certainty carries through to the YPR
model and is demonstrated by the dramatic increase in certainty from
the herring YPR model compared to cod. In the survey data
the age of the fish is recorded as the
FIGURE 2 The von Bertalanffy
growth function fitted to simulated data
with ai =ti +qi. The violin plots (Hintze &
Nelson, 1998) show the marginal posterior
distributions for each of the parameters
fitted to different sample sizes. TABLE 2 The posterior mean and
standard deviation of the parameters of
the VBGF, the percentage increase in YPR
with fishing at Fmax, relative to model II,
and the WAIC for the different models
fitted to herring data 3.2 | Herring We compared three Increasing the amount of data increases certainty in the param-
eters of VBGF, which in turn, can increase certainty in models used
by management and policymakers. This can be seen as we get lower 7064 |
FIGURE 3 The line shows the fitted von Bertalanffy growth
function for the posterior mean, and the points show the posterior
mean age of each herring when it was surveyed from model IV. The
dotted lines show the 90% posterior predictive credible interval
12
10
8
6
4
2
0
50
100
150
200
250
300
350
400
Age (years)
Length (mm) 7064 |
12
10
8
6
4
2
0
50
100
150
200
250
300
350
400
Age (years)
Length (mm) SPENCE and TURTLE 7064 4.1 | Ig The simulation study showed that ignoring age uncertainty leads to
inconsistent models both for fish spawning according to the herring
spawning distribution and that of cod. For both herring and cod, fitting
the VBGF by treating the ages as known, model I gave much lower k
and larger l∞ values than the other models. In reality, fish caught in the
fourth quarter are only a quarter of a year younger than if they were
caught in the first quarter of the following year’s survey; however in
the data, they would be a year apart resulting in much overlap of fish
between ages. This, therefore, suggests a lower growth rate, k, larger
l∞ values, as these two parameters are negatively correlated (Siegfried
& Sansó, 2006), and larger errors, σ. An alternative is to increase the age of fish by the quarter that they
were caught in as described in Sparre & Venema, (1998). In practice,
this means that fish surveyed in quarter 4 have their age increased by
0.75. For fish with a short-spawning period, the difference between
ages of fish in the data and the truth will be small and centered on zero,
however for fish with large-spawning times, the difference between
the data and the truth, although still centered on 0, will be much larger. FIGURE 3 The line shows the fitted von Bertalanffy growth
function for the posterior mean, and the points show the posterior
mean age of each herring when it was surveyed from model IV. The
dotted lines show the 90% posterior predictive credible interval FIGURE 3 The line shows the fitted von Bertalanffy growth 4.2 TABLE 3 The posterior mean and
standard deviation of the parameters of
the VBGF, the percentage increase in YPR
with fishing at Fmax, relative to model II,
and the WAIC for the different models
fitted to cod data VBGF, von Bertalanffy growth function; WAIC, Watanabe-Akaike information criterion; YPR,
yield-per-recruit. length at a given age. This improved the fitting of the model for both
cod and herring, as shown by lower values of WAIC, than other mod-
els. The improvement is greater for herring, almost three times better
(in terms of variance) than the method described in Sparre & Venema,
(1998), than in cod, 1.15 times better. We suspect that this may be
because of the larger spawning season allowing the ages of the fish to
vary more thus fitting better. length at a given age. This improved the fitting of the model for both
cod and herring, as shown by lower values of WAIC, than other mod-
els. The improvement is greater for herring, almost three times better
(in terms of variance) than the method described in Sparre & Venema,
(1998), than in cod, 1.15 times better. We suspect that this may be
because of the larger spawning season allowing the ages of the fish to
vary more thus fitting better. We found that cod had done about three quarters of their annual
growth by the beginning of the 4th quarter. This meant that the poste-
rior distributions of models II and III were very similar. Conversely, by
the beginning of the fourth quarter, herring had already grown about
90% of their annual growth rate. This had an effect on the marginal
posterior distribution of k. In model V, we have N fish all aged differently, so we required f(x)
to be explicitly defined for x∈[0,1). We found that, for all but very sim-
ple f(⋅)s, more information about the growth rate was required, such The models that included different spawning times, models IV and
V, suggested large differences in the posterior distribution for cod de-
spite the spawning season being short compared to that of herring. This is because of large variations in the lengths found in cod aged 1
in quarter 1. 4.2 In the model with ai =ti +qi, this variation is taken into ac-
count by increasing σ, whereas in the spawning model, the largest cod
caught in quarter 1 having survived 1 winter could be up to half a year
older than the smallest fish in the same category, as shown in Figure 5,
suggesting higher values of k. FIGURE 5 The spawning times of a cod of length 120 mm and a
cod of length 340 mm. Both were surveyed in quarter 1 and have age
1 in the survey data. The solid line is the spawning distribution of the
whole population
0.0
0.2
0.4
0.6
0.8
1.0
5
4
3
2
1
0
Proportion of Year
Density
120 mm
340 mm
Spawning dist. 0.0
0.2
0.4
0.6
0.8
1.0
5
4
3
2
1
0
Proportion of Year
Density
120 mm
340 mm
Spawning dist. TABLE 3 The posterior mean and
standard deviation of the parameters of
the VBGF, the percentage increase in YPR
with fishing at Fmax, relative to model II,
and the WAIC for the different models
fitted to cod data 4.2 different ways of combining the data from multiple surveys and
showed that the values of the VBGF parameters are sensitive to
the model chosen. In order to try and account for this we introduced a version of the
VBGF that models the spawning time of individuals as well as their FIGURE 4 Residual analysis for model
IV fitted to the herring data and simulated
herring data
Herring data
−4
−2
4
2
0
2
0
12
10
8
6
4
2
12
10
8
6
4
2
Simulated data
−4
−2
4
ti + qi
Standardized residuals Herring data
−4
−2
4
2
0
12
10
8
6
4
2
Standardized residuals Simulated data Standardized residuals FIGURE 4 Residual analysis for model
IV fitted to the herring data and simulated
herring data FIGURE 4 Residual analysis for model
IV fitted to the herring data and simulated
herring data | SPENCE and TURTLE 7065 Model
I
II
III
IV
V
k
Mean
0.04
0.18
0.18
0.24
0.24
SD
0.013
0.006
0.006
0.011
0.011
l∞
Mean
4,166.7
1,374.2
1,384.6
1,148.3
1,143.2
SD
1,631.21
35.23
35.16
30.83
30.55
t0
Mean
−0.954
−0.004
−0.004
−0.165
−0.179
SD
0.057
0.004
0.004
0.020
0.020
σ
Mean
0.209
0.149
0.149
0.139
0.139
SD
0.00276
0.00209
0.00195
0.00195
0.00203
% increase YPR
Mean
48
0
0
3
2
SD
11
0
3
3
2
WAIC
−810
−2,717
−2,720
−2,919
−2,945
VBGF, von Bertalanffy growth function; WAIC, Watanabe-Akaike information criterion; YPR,
yield-per-recruit. TABLE 3 The posterior mean and
standard deviation of the parameters of
the VBGF, the percentage increase in YPR
with fishing at Fmax, relative to model II,
and the WAIC for the different models
fitted to cod data Model
I
II
III
IV
V
k
Mean
0.04
0.18
0.18
0.24
0.24
SD
0.013
0.006
0.006
0.011
0.011
l∞
Mean
4,166.7
1,374.2
1,384.6
1,148.3
1,143.2
SD
1,631.21
35.23
35.16
30.83
30.55
t0
Mean
−0.954
−0.004
−0.004
−0.165
−0.179
SD
0.057
0.004
0.004
0.020
0.020
σ
Mean
0.209
0.149
0.149
0.139
0.139
SD
0.00276
0.00209
0.00195
0.00195
0.00203
% increase YPR
Mean
48
0
0
3
2
SD
11
0
3
3
2
WAIC
−810
−2,717
−2,720
−2,919
−2,945
VBGF, von Bertalanffy growth function; WAIC, Watanabe-Akaike information criterion; YPR,
yield-per-recruit. AUTHOR CONTRIBUTIONS MAS and AJT conceived the ideas and designed the methodology;
AJT extracted the data; MAS analyzed the data; MAS led the writing
of the manuscript. All authors contributed critically to the drafts and
gave final approval for publication. 4.5 | Future work Throughout this study we assumed multiplicative log-normal errors
in order to describe the uncertainty not captured by VBGF. This pro-
vides several advantages, such as the interpretations of li and reduced
standard errors (Quintero, Contreras-Reyes, Wiff, & Arellano-Valle,
2017), however other, more flexible, error distributions that have
been proposed, such as the log-skewed-t distribution (e.g., Contreras-
Reyes & Arellano-Valle, 2013). Furthermore, we plan to investigate
whether fish spawned in the same time period (e.g., year) have similar
growth patterns in order to try and explain some of the uncertainty
not captured in our model. ACKNOWLEDGMENTS MAS was supported by the Natural Environment Research Council
and Department for Environment, Food and Rural Affairs [grant
number NE/L003279/1, Marine Ecosystems Research Programme],
and AJT was supported by Natural Environment Research Council
[grant number NE/L002450/1, NERC Research Experience
Placements]. The authors would like to thank Paul G. Blackwell,
Tom Webb, Toni I. Gossman, Briony Norton, Fredrick Heather, and
Christopher Griffiths for additional advice as well as two anony-
mous reviews. As shown in this article, fitting to different quarters of data can
lead to different posterior distributions of the VBGF. In the Supporting
Information, we demonstrate that for cod sampling from one quarter
only can lead to inconsistent models however for herring it does not. We speculate that this is because of the larger spawning season and,
for one survey, you get a greater range of ages of fish and therefore
more information. Further research will investigate the time and the
gear used to perform the survey and their effect on the VBGF (Wilson
et al., 2015). 4.4 | Yield-per-recruit Yield-per-recruit analysis often provides reference points for manage-
ment purposes (Katsukawa, 2005). We demonstrated the importance
of quantifying the uncertainty in the estimates of the VBGF as the out-
come of the YPR analysis is also uncertain regardless of which model
was used to find the parameters. Furthermore, the YPR analysis is sen-
sitive to the model used to fit the VBGF, and therefore, the “optimal”
strategy which a policymaker decides can also be sensitive to this. as surveys from other quarters, and therefore we set f(⋅) to be a linear
interpolator. We found that this did not have a large effect on the
parameters of the VBGF for either herring or cod, suggesting that our
model is robust to seasonal growth rates. DATA ACCESSIBILITY The data and the R scripts for this study can be found in the Supporting
Information. Our model is not just specific to fish or the VBGF but can also be
used in any area of ecology where the growth curves are fitted to data
when the age of an individual is not known exactly; such as in studies
of elephants (Shrader et al., 2006; Trimble et al., 2011), brown bears
(Zedrosser, Bellemain, Taberlet, & Swenson, 2007), and birds (Tjørve &
Tjørve, 2010), just to name a few. 4.6 | Summary Generally, if the true age of an individual was known, the growth
curve would be fitted with this as opposed to the number of
winters that the individual has survived (e.g., Nurdin, Sondita,
Yusfiandayani, & Baskoro, 2016). As the ages of fish are binned
in the survey data, the true age of fish is uncertain. By treating
the ages in the survey data as the truth, either exactly or adjust-
ing for the quarter caught, this uncertainty is ignored, which, as
shown in this study, the fitted parameter values can be sensitive to. We described here a model that treats the true ages as uncertain
and infers them, both from the survey data and spawning data. The
model is fitted as if we knew the true age of the individual, which
is what would happen if we did, and includes uncertainty as we do
not know the true age of the fish. This, therefore, is a more robust
way of fitting growth curves. Furthermore, our model decreases
the uncertainty in models used by policymakers which means that
they can be more confident when making their management strate-
gies (Harwood & Stokes, 2003). None declared. In this study, we also introduced a new way of describing sea-
son VBGF. Further research could investigate what effect f(⋅) has
on the posterior distributions, especially for species that may only
have short growth seasons. As long as f(0)=0 and f(1)=1, k and l∞
still have the same interpretation as in equation (1). We addition-
ally suggest that f′(x)≥0, which would mean that an individual never
shrinks, and that it is circular, that is f (0)=f (1) and similarly for
higher derivatives. Unlike the linear interpolator, Somers (1988)’s
model is circular but is not particularly flexible (Pauly et al., 1992). A more flexible function could be the cumulative distribution of the
von Mises distribution. 4.3 | Seasonal growth Growth does not necessarily follow the VBGF throughout the year
but can be seasonal (García-Berthou et al., 2012). To test what ef-
fect this had on the parameters, we allowed the growth rate to
change over the year. By explicitly describing how the growth is
divided over a single year, we give a general way of specifying the
seasonal growth of an individual. Although there are a number of
models which would enable us to describe the seasonal growth
rates (e.g., Somers, 1988), we were interested in the VBGF param-
eters rather than predicting the size of an individual in the future,
so we only require f(x), the proportion of annual growth by x, to
be defined for values of x in the study. For model III, where fish
were assumed to have spawned on January 1st, x was either 0 or
0.75. f(0)=0 by definition so by adding an additional parameter
f(0.75)=c2, we are not implying any functional form on f(⋅) except
that f(0.75)=c2. FIGURE 5 The spawning times of a cod of length 120 mm and a
cod of length 340 mm. Both were surveyed in quarter 1 and have age
1 in the survey data. The solid line is the spawning distribution of the
whole population SPENCE and TURTLE 7066 CONFLICT OF INTEREST None declared. Austin, S. H., Robinson, T. R., Robinson, W. D., & Ricklefs, R. E. (2011). Potential biases in estimating the rate parameter of sigmoid growth
functions. Methods in Ecology and Evolution, 2, 43–51. Hintze, J. L., & Nelson, R. D. (1998). Violin plots: A box plot-density trace
synergism. The American Statistician, 52(2), 181–184. Hoenig, J. M. (1983). Empirical use of longevity data to estimate mortality
rates. Fishery Bulletin, 82, 898–903. von Bertalanffy, L. (1957). Quantitative laws in metabolism and growth. The
Quarterly Review of Biology, 32, 217–231. Hoffman, M. D., & Gelman, A. (2014). The No-U-turn sampler: Adaptively
setting path lengths in Hamilitonian Monte Carlo. Journal of Machine
Learning Research, 15, 1351–1381. Best, D. J., & Fisher, N. I. (1979). Efficient simulation of the von Mises dis-
tribution. Applied Statistics, 28, 152–157. ICES Database of Trawl Surveys (DATRAS), Extraction 12 MAY (2016). International Bottom Trawl Survey (IBTS) data 2000-2016; http://da-
tras.ices.dk. ICES, Copenhagen. Beverton, R., & Holt, S. (1959). A review of the lifespan and mortality rates
of fish in nature and their relationship to growth and other physiolog-
ical characteristics. Ciba Foundation Symposium-The Lifespan of Animals
(Colloquia on Ageing), 5, 142–180. ICES (2016a). Report of the Herring Assessment Working Group for the Area
South of 62°N (HAWG). Tech. rep., ICES, Copenhagen. Blanchard, J. L., Andersen, K. H., Scott, F., Hintzen, N. T., Piet, G., & Jennings,
S. (2014). Evaluating targets and trade-offs among fisheries and con-
servation objectives using a multispecies size spectrum model. Journal
of Applied Ecology, 51(3), 612–662. ICES (2016b). Report of the Working Group on Celtic Seas Ecoregion (WGCSE). Tech. rep., ICES, Copenhagen. Katsukawa, T. (2005). Evaluation of current and alternative fisheries man-
agement scenarios based on spawning-per-recruit (SPR), revenue-
per-recruit (RPR), and yield-per-recruit (YPR) diagrams. ICES Journal of
Marine Science: Journal du Conseil, 62(5), 841–846. Brander, K. (1994). The location and timing of cod spawning around the
British Isles. ICES Journal of Marine Science, 51(1), 71–89. Chambers, M. S., Sidhu, L. A., O’Neil, B., & Sibanda, N. (2017). Flexible von
Bertalanffy growth models incorporating Bayesian splines. Ecological
Modelling, 355, 1–11. Knijn, R. J., Boon, T. W., Heessen, H. J. L., & Hislop, J. R. G. (1993). Atlas of
North Sea fishes: ICES Cooperative Research Report No. 194. Tech. rep.,
ICES, Copenhagen. Chen, Y., Jackson, D. A., & Harvey, H. H. (1992). A comparison of von
Bertalanffy and polynomial functions in modelling fish growth data. Canadian Journal of Fisheries and Aquatic Sciences, 49, 1228–1235. Lorenzen, K. (2000). Allometry natural mortality as a basis for assessing
optimal release size in fish-stocking programmes. Canadian Journal of
Fisheries and Aquatic Sciences, 57, 2374–2381. Contreras-Reyes, J. E., & Arellano-Valle, R. B. (2013). Growth estimates of
cardinalfish (Epigynous crassicaudus) based on scale mixtures of skew-
normal distributions. Fisheries Research, 147, 137–144. Nelson, G. A. (2016). fishmethods: Fishery science methods and models in R. Retrieved from https://CRAN.R-project.org/package=fishmethods Cope, J., & Punt, A. E. (2007). Admitting ageing error when fitting growth
curves: An example using the von Bertalanffy growth function with
random effects. Canadian Journal of Fisheries and Aquatic Sciences, 64,
205–218. Nurdin, E., Sondita, M. F. A., Yusfiandayani, R., & Baskoro, M. S. (2016). Growth and mortality parameters of yellowfin tuna (Thunnus albacares)
in Palabuhanratu waters, west Java (eastern Indian Ocean). Aquaculture,
Aquarium, Conservation and Legislation-International Journal of the
Bioflux Society (AACL Bioflux), 9(3), 741–747. Datta, S., & Blanchard, J. L. (2016). The effects of seasonal processes on
size spectrum dynamics. Canadian Journal of Fisheries and Aquatic
Sciences, 73(4), 598–610. Paine, C. E. T., Marthews, T. R., Vogt, D. R., Purves, D., Rees, M., Hector, A.,
& Turnbull, L. A. (2012). How to fit nonlinear plant growth models and
calculate growth rates: An update for ecologists. Methods in Ecology and
Evolution, 3, 245–256. Doll, J. C., Lauer, T. E., & Clark-Kolaks, S. (2017). Yield-per-recruit model-
ling of two piscivores in a Midwestern reservoir: A Bayesian approach. Fisheries Research, 191, 200–210. Pardo, S. A., Cooper, A. B., & Dulvy, N. K. (2013). Avoiding fishy growth
curves. Methods in Ecology and Evolution, 4, 353–360. Einum, S., Forseth, T., & Finstad, A. G. (2012). Individual variation in response
to intraspecific competition: Problems with inference from growth vari-
ation measures. Methods in Ecology and Evolution, 3, 438–444. Pauly, D. (1980). On interrelationships between natural mortality,
growth parameters and mean environmental temperature in 175 fish
stocks. Journal du Conseil International pour l’Exploration de la Mer, 39,
175–192. Essington, T. E., Kitchell, J. F., & Walters, C. J. (2001). The von
Bertalanffy growth function, bioenergetics, and the consumption
rate of fish. Canadian Journal of Fisheries and Aquatic Sciences, 58,
2129–2138. Pauly, D., Soriano-Bartz, M., Moreau, J., & Jarre-Teichmann, A. (1992). A new model accounting for seasonal cessation of growth in fishes. Marine and Freshwater Research, 43, 1151–1156. Froese, R. and Pauly, D. Editors. (2016). FishBase. REFERENCES Andersen, K. H., & Beyer, J. E. (2006). Asymptotic size determines species
abundance in marine size spectrum. The American Naturalist, 168(1),
54–61. SPENCE and TURTLE 7067 Zedrosser, A., Bellemain, E., Taberlet, P., & Swenson, J. (2007). Genetic esti-
mates of annual reproductive success in male brown bears: The effects
of body size, age, internal relatedness and population density. Journal
of Animal Ecology, 76(2), 368–375. Thorpe, R. B., Le Quesne, W. J. F., Luxford, F., Collie, J. S., & Jennings,
S. (2015). Evaluation and management implications of uncertainty
in a multi-species size-structured model of population and com-
munity responses to fishing. Methods in Ecology and Evolution, 6(1),
49–58. Tjørve, K. M. C., & Tjørve, E. (2010). Shape and functions of bird-growth mod-
els: How to characterise chick postnatal growth. Zoology, 113, 326–333. World Wide Web elec-
tronic publication. www.fishbase.org, version (12/2016). Quintero, F. O. L., Contreras-Reyes, J. E., Wiff, R., & Arellano-Valle, R. B. (2017). Flexible Bayesian analysis of the von Bertanaffy growth func-
tion with the use of a log-skew-t distribution. Fishery Bulletin, 115(1),
13–26. Gabriel, W. L., Sissenwine, M. P., & Overholtz, W. J. (1989). Analysis of
spawning stock biomass per recruit: An example for Georges Bank
haddock. North American Journal of Fisheries Management, 9, 383–391. R Core Team (2015). R: A language and environment for statistical comput-
ing. Vienna, Austria: R Foundation for Statistical Computing. Retrieved
from https://www.R-project.org/ García-Berthou, E., Carmona-Catot, G., Merciai, R., & Ogle, D. H. (2012). A
technical note on seasonal growth models. Reviews in Fish Biology and
Fisheries, 22(3), 635–640. Schnute, J., & Fournier, D. (1980). A new approach to length-frequency
analysis: Growth structure. Canadian Journal of Fisheries and Aquatic
Sciences, 37, 1337–1351. Gelman, A., Lee, D., & Guo, J. (2015). Stan: A probabilistic programming lan-
guage for Bayesian inference and optimization. Journal of Educational
and Behavioural Statistics, 40, 530–543. Gelman, A., & Rubin, D. B. (1992). Inference from iterative simulation using
multiple sequences. Statistical Science, 7, 457–511. Shrader, A. M., Ferreira, S. M., McElveen, M. E., Lee, P. C., Moss, C. J., & van
Aarde, R. J. (2006). Growth and age determination of African savanna
elephants. Journal of Zoology, 270, 40–48. Gislason, H., Daan, N., Rice, J. C., & Pope, J. G. (2010). Size, growth, tem-
perature and the natural mortality of marine fish. Fish and Fisheries, 11,
149–158. Siegfried, K. I., & Sansó, B. (2006). Two Bayesian methods for estimating
parameters of the von Bertalanffy growth equation. Environmental
Biology of Fishes, 77, 301–308. Hamel, O. S. (2015). A method for calculating a meta-analytical prior for the
natural mortality rate using multiple life history correlates. ICES Journal
of Marine Science, 72, 62–69. Somers, I. F. (1988). On seasonally oscillating growth function. Fishbyte,
6, 8–11. Harwood, J., & Stokes, K. (2003). Coping with uncertainty in ecological
advice: Lessons from fisheries. Trends in Ecology and Evolution, 18(12),
617–622. Sparre, P., & Venema, S. C. (1998). Introduction to Tropical Fish Stock
Assessment: Manual. Tech. Rep. FAO Fisheries Technical Paper 306/1
Rev. 2, Food and Agriculture Organization of the United Nations, Rome 7068 SPENCE and TURTLE SUPPORTING INFORMATION Trimble, M. J., van Aarde, R. J., Ferreira, S. M., Nørgaard, C. F., Fourie, J.,
Lee, P. C., & Moss, C. J. (2011). Age determination by back length for
african savanna elephants: Extending age assessment techniques for
aerial-based surveys. PLoS ONE, 6(10), 1–7. Additional Supporting Information may be found online in the
supporting information tab for this article. Walters, C. J., & Martell, S. J. D. (2004). Fisheries ecology and management. Princeton, NJ: Princeton University Press. Watanabe, S. (2010). Asymptotic equivalence of Bayes cross validation
and widely applicable information criterion in singular learning theory. Journal of Machine Learning Research, 11, 3571–3594. How to cite this article: Spence MA, Turtle AJ. Making the
most of survey data: Incorporating age uncertainty when
fitting growth parameters. Ecol Evol. 2017;7:7058–7068. https://doi.org/10.1002/ece3.3280 How to cite this article: Spence MA, Turtle AJ. Making the
most of survey data: Incorporating age uncertainty when
fitting growth parameters. Ecol Evol. 2017;7:7058–7068. https://doi.org/10.1002/ece3.3280 Wilson, K. L., Matthias, B. G., Barbour, A. B., Ahrens, R. N. M., Tuten, T.,
& Allen, M. S. (2015). Combining samples from multiple gears helps
to avoid fishy growth curves. North American Journal of Fisheries
Management, 35, 1121–1131.
|
https://openalex.org/W2903406976
|
https://jurnal.untirta.ac.id/index.php/JRBM/article/download/3832/2792
|
Indonesian
| null |
PENGARUH MOTIVASI INTRINSIK, KOMUNIKASI INTERPERSONAL DAN KEPEMIMPINAN TRANSFORMASIONAL TERHADAP KEPUASAN KERJA DALAM MENINGKATKAN KINERJA PEGAWAI (Kantor Kementerian Agama Kota Serang)
|
Jurnal Riset Bisnis dan Manajemen Tirtayasa
| 2,018
|
cc-by-sa
| 9,891
|
PENGARUH MOTIVASI INTRINSIK, KOMUNIKASI
INTERPERSONAL DAN KEPEMIMPINAN TRANSFORMASIONAL
TERHADAP KEPUASAN KERJA DALAM
MENINGKATKAN KINERJA PEGAWAI
(Kantor Kementerian Agama Kota Serang) Rohmat Hidayat, Roni Kambara, Lutfi
Program Pascasarjana Magister Manajemen
Universitas Sultan Ageng Tirtayasa
rohmat7hidayat@gmail.com Rohmat Hidayat, Roni Kambara, Lutfi
Program Pascasarjana Magister Manajemen
Universitas Sultan Ageng Tirtayasa
rohmat7hidayat@gmail.com Abstract The purpose of this study were to describe the effect of intrinsic motivation, interpesonal
communication and transformational leadership on job satisfaction to improve employee
performance. This reserch consist of five variables, these variables were tree independent
variables (exogen), one dependent variable (endogen), and one intervening variable. The
three independent variables were intrinsic motivation, interpesonal communication and
transformational leadership. Whereas, the dependent variable was employee performance,
and the intervening variable was job satisfaction. This study was conducted at the Office of
Ministry of Religion in Serang Banten. The purposive sampling technique was used based
on judgement sampling to 81 respondents. The primary data was collected through checklist
and open-ended questioner. Data were analyzed by Structural Equation Modelling
technique using SmartPLS software. There six of Seven hypotheses that have been suported
or accepted: intrinsic motivation positive and significant impact on job satisfaction,
interpesonal communication positive and significant impact on job satisfaction,
transformational positive and significant impact on job satisfaction, intrinsic motivation
positive and significant impact on employee performance, transformational leadership
positive and significant impact on employee performance, and job satisfaction positive and
significant impact on employee performance. While there was one hypothesis is unsupported
or rejected: interpesonal communication positive but insignificant impact on employee
performance. Keyword: intrinsic motivation, interpesonal communication, transformational, leadership,
job satisfaction, employee performance. Keyword: intrinsic motivation, interpesonal communication, transformational, leadership,
job satisfaction, employee performance. Pendahuluan merupakan penggerak dan pengelola faktor-
faktor produksi seperti modal,
bahan
mentah, peralatan dan lain-lain dalam upaya
mencapai tujuan organisasi. Sumber daya manusia merupakan
salah satu unsur penting dalam upaya
pencapaian tujuan dan keberhasilan suatu
organisasi sebagaimana yang dikemukakan
oleh Dessler (2011:5) Sumber daya manusia Organisasi yang ditunjang oleh pegawai
dengan tingkat kinerja yang tinggi dalam
melaksanakan setiap tugasnya, akan mampu
bersaing
secara
kompetitif
dan
terus
mengalami perkembangan ke arah yang
lebih
baik. Sedarmayanti
(2013:260)
menjelaskan kinerja adalah sebuah proses
manajemen
yang
menyeluruh
dimana Jurnal Riset Bisnis dan Manajemen Tirtayasa
(JRBMT), Vol. 2 (1): hh.43-66 (Juni 2018)
ISSN (Online) 2599-0837,
http://jurnal.untirta.ac.id/index.php/JRBM
© 2018 Magister Manajemen UNTIRTA Jurnal Riset Bisnis dan Manajemen Tirtayasa
(JRBMT), Vol. 2 (1): hh.43-66 (Juni 2018)
ISSN (Online) 2599-0837,
http://jurnal.untirta.ac.id/index.php/JRBM
© 2018 Magister Manajemen UNTIRTA 43 Jurnal Riset Bisnis dan Manajemen Tirtayasa hasilnya dapat diukur dan dibuktikan secara
nyata. Sedangkan
Handoko
(2010)
menyatakan bahwa kinerja adalah hasil yang
dapat
dicapai
oleh
seseorang
atau
sekelompok orang dalam pelaksanaan tugas,
pekerjaan yang baik, artinya mencapai
sasaran atau standar kerja yang telah
ditentukan sebelumnya dan atau bahkan
dapat melebihi standar yang telah ditentukan
oleh perusahaan dalam periode tertentu. Lebih lanjut Rivai dan Basri (2010:50)
menyatakan kinerja adalah hasil atau tingkat
keberhasilan seseorang secara keseluruhan
selama periode tertentu dalam melaksanakan
tugas
dibandingkan
dengan
berbagai
kemungkinan, seperti standar hasil kerja,
target atau sasaran atau kriteria yang telah
ditentukan terlebih dahulu telah disepakati
bersama. memeriksa/mengkaji
dan
mengevaluasi
secara berkala kinerja seseorang. memeriksa/mengkaji
dan
mengevaluasi
secara berkala kinerja seseorang. Dalam institusi pemerintah, termasuk di
Kantor Kementerian Agama Kota Serang
ketentuan tentang penilaian kerja bagi
Pegawai Negeri Sipil (PNS) tercantum
dalam Peraturan Pemerintah (PP) No. 46
tahun 2011. Tujuan penilaian kinerja adalah
untuk menjamin objektivitas pembinaan
pegawai yang dilakukan berdasarkan sistem
prestasi kerja dan sistem karier yang
dititikberatkan pada sistem prestasi kerja. Dalam peraturan pemerintah ini yang
dimaksud dengan prestasi kerja pegawai
adalah hasil kerja yang dicapai oleh setiap
pegawai pada satuan organisasi sesuai
dengan sasaran kerja pegawai dan perilaku
kerja. Tolak ukur penilaian kinerja pegawai
terdiri dari unsur sasaran kinerja pegawai
atau disingkat SKP dan perilaku kerja, SKP
adalah rencana kerja dan target yang akan
dicapai oleh seorang pegawai yang terdiri
dari aspek kuantitas, kualitas, waktu, dan
biaya. Pendahuluan Sedangkan perilaku kerja adalah
setiap tingkah laku, sikap atau tindakan yang
dilakukan
oleh
pegawai
atau
tidak
melakukan
sesuatu
yang
seharusnya
dilakukan sesuai dengan ketentuan peraturan
perundang-undangan yang terdiri dari aspek
orientasi pelayanan, integritas, komitmen,
disiplin, kerja sama, dan kepemimpinan. Kantor Kementerian Agama Kota
Serang merupakan salah satu organisasi
pemerintah
yang
menyelenggarakan
pelayanan publik di bidang keagamaan yang
berada di wilayah Kota Serang dalam hal ini
menguasai hajat hidup orang banyak, untuk
itu dituntut untuk memberikan pelayanan
yang maksimal terhadap semua pihak yang
berkepentingan. Kantor
Kementerian
Agama Kota Serang harus mampu melayani
masyarakat
sebaik
mungkin
sehingga
mereka
benar-benar
merasakan
puas
terhadap pelayanan yang diberikan. Semua
itu dapat terjadi manakala pegawainya
memiliki kinerja yang baik, sehingga
mampu
memberikan
pelayanan
yang
maksimal kepada masayarakat. Upaya untuk meningkatkan kinerja
pegawai bukanlah hal yang sederhana, sebab
kinerja pegawai bukan merupakan konsep
yang berdiri sendiri, melainkan tergantung
kepada sejumlah variabel atau faktor-faktor
yang memengaruhinya. Persoalan kinerja
pegawai
akan
dapat
terlaksana
dan
terpecahkan apabila faktor-faktor tersebut
dapat dikelola dan dipelihara dengan baik. Martoyo (2010:15) mengungkapkan bahwa
faktor yang berpengaruh terhadap kinerja
antara lain motivasi, kepuasan kerja,
lingkungan
kerja,
sIstem
kompensasi,
kepemimpinan
dan
perilaku
lainnya. Nitisemito (1996) dalam Tika (2010)
menyebutkan bahwa faktor-faktor yang
dapat memengaruhi kinerja antara lain
insentif,
pendidikan
dan
pelatihan,
kepemimpinan,
absensi,
komunikasi, Untuk mengetahui kinerja pegawai
dalam setiap organisasi dapat diukur dengan
menggunakan metode tertentu. Seperti yang
dikemukakan Tika (2010) kinerja setiap unit
organisasi harus diukur dengan metode
statistik, khususnya tentang mutu suatu
produksi. Dalam standar pengukuran kinerja
perlu dirumuskan untuk dijadikan indikator
perbandingan
antara
apa
yang
telah
dihasilkan dengan apa yang diharapkan,
kaitannya dengan pekerjaan yang telah
dikerjakan
seseorang. Menurut
Sedarmayanti
(2013:261)
pengukuran
kinerja
adalah
sistem
formal
untuk 44 JRBM Tirtayasa Volume 2 (1) - Juni 2018 motivasi
dan
lingkungan
kerja. Dari
beberapa upaya untuk meningkatkan kinerja
pegawai sebagaimana menurut para ahli di
atas, yang menjadi fokus dalam penelitian
ini adalah; 1) faktor motivasi, 2) faktor
komunikasi
interpersonal,
3)
faktor
kepemimpinan transformasional, dan 4)
faktor kepuasan kerja. dalam diri pekerja sebagai individu berupa
kesadaran
mengenai
pentingnya
atau
manfaat
atau
makna
pekerjaan
yang
dilaksanakannya. Dengan kata lain motivasi
ini
bersumber
dari
pekerjaan
yang
dilaksanakannya
baik
karena
mampu
memenuhi kebutuhan atau menyenangkan
atau memungkinkan mencapai suatu tujuan
maupun
karena
memberikan
harapan
tertentu yang positif di masa depan. Pertama, faktor motivasi. Sebuah
organisasi harus memerhatikan bagaimana
menjaga dan mengelola motivasi pegawai
dalam bekerja agar selalu fokus pada tujuan
perusahaan. Pendahuluan 45 Jurnal Riset Bisnis dan Manajemen Tirtayasa Kemudian menurut Robbins dan Judge
(2015)
bahwa
kepemimpinan
adalah
kemampuan untuk memengaruhi kelompok
guna mencapai sebuah visi atau serangkaian
tujuan yang ditetapkan. Salah satu tipe
pendekatan
kepemimpinan
adalah
kepemimpinan transformasional. Menurut
Robbins
dan
Judge
(2015:90)
kepemimpinan
transformasional
adalah
pemimpin
yang
menginspirasi
para
pengikutnya
untuk
mengenyampingkan
kepentingan pribadi mereka dan memiliki
kemampuan memengaruhi yang luar biasa. Kemudian menurut Robbins dan Judge
(2015)
bahwa
kepemimpinan
adalah
kemampuan untuk memengaruhi kelompok
guna mencapai sebuah visi atau serangkaian
tujuan yang ditetapkan. Salah satu tipe
pendekatan
kepemimpinan
adalah
kepemimpinan transformasional. Menurut
Robbins
dan
Judge
(2015:90)
kepemimpinan
transformasional
adalah
pemimpin
yang
menginspirasi
para
pengikutnya
untuk
mengenyampingkan
kepentingan pribadi mereka dan memiliki
kemampuan memengaruhi yang luar biasa. dikategorikan sangat
baik dibandingkan
dengan tahun sebelumnya, yaitu pada tahun
2016 ebesar 15.32%, pada tahun 2015
sebesar 18.92%, dan pada tahun 2014
sebesar 19.64%. Pendahuluan Menjaga motivasi karyawan
sangat penting karena motivasi itu adalah
salah satu motor penggerak bagi setiap
individu yang mendasari mereka untuk
bertindak dan melakukan sesuatu. Orang
tidak akan melakukan sesuatu hal secara
optimal apabila tidak mempunyai motivasi
yang tinggi dari dalam dirinya sendiri untuk
melalukan hal tersebut. Robin dan Judge
(2015:222) mendefinisikan motivasi sebagai
suatu proses yang menjelaskan intensitas,
arah dan ketekunan seorang individu untuk
mencapai tujuannya. Definisi lain motivasi
pemberian
daya
penggerak
yang
menciptakan kegairahan kerja seseorang
agar mau bekerja sama, bekerja efektif, dan
terintegrasi dengan segala daya upayanya
untuk
mencapai
kepuasan
(Hasibuan,
2010:143). Dari definisi tersebut dapat
dicermati bahwa motivasi menjadi bagian
penting yang mendasari individu atau
seseorang dalam melakukan sesuatu atau
mencapai tujuan tertentu yang diinginkan. Kedua, faktor komunikasi. Menurut
Ardana at al. (2012:138) komunikasi
merupakan kunci pembuka dapat terjadinya
hubungan
kerjasama
antara
pimpinan
dengan karyawan dan antar karyawan itu
sendiri. Handoko (2010:272) menjelaskan
bahwa
komunikasi
merupakan
proses
memberikan pemahaman berupa ide atau
informasi kepada seseorang. Komunikasi
yang baik akan membantu karyawan dalam
memahami
pekerjaannya
secara
jelas
sehingga
perlahan
kinerjanya
akan
meningkat. Proses pertukaran informasi di
antara seseorang dengan paling kurang
seorang lainnya atau biasanya di antara dua
orang yang dapat langsung diketahui
balikannya
disebut
komunikasi
interpersonal (Arni, 2011). Sementara itu
menurut De Vito dalam Effendy (2010: 23),
komunikasi
interpersonal
adalah
penyampaian pesan oleh satu orang dan
penerimaan pesan oleh orang lain atau
sekelompok kecil orang, dengan berbagai
dampaknya dan dengan peluang untuk
memberikan umpan balik segera. p
g
Ketiga,
faktor
kepemimpinan
transformasional. Kepemimpinan
dibutuhkan
manusia
karena
adanya
keterbatasan-keterbatasan tertentu pada diri
manusia. Dari sinilah timbul kebutuhan
untuk memimpin dan dipimpin. Menurut
Rivai at al. (2014) kepemimpinan adalah
proses memengaruhi atau memberi contoh
kepada pengikut-pengikutnya lewat proses
komunikasi dalam upaya mencapai tujuan
organisasi. Kepemimpinan adalah proses
mengarahkan,
membimbing
dan
memengaruhi pikiran, perasaan, tindakan
dan
tingkah
laku
orang
lain
untuk
digerakkan
ke
arah
tujuan
tertentu. Berdasarkan
aktivasi
perilaku,
menurut Herzberg’s Two Factors Theory
(1966) dalam Tan dan Waheed (2011)
motivasi
dibagi
menjadi
dua
faktor
diantaranya faktor motivator atau yang
sering disebut dengan motivasi intrinsik dan
faktor hygiene atau yang sering disebut
dengan
motivasi
ekstrinsik. Motivasi
intrinsik adalah sesuatu yang datang dari
dalam diri seseorang sedangkan motivasi
ekstrinsik adalah motivasi yang disebabkan
oleh faktor eksternal atau faktor dari luar diri
seseorang
(Tan
dan
Waheed,
2011). Menurut Nawawi (2010) motivasi intrinsik
adalah pendorong kerja yang bersumber dari Ketiga,
faktor
transformasional. Kepemimpinan Transformasional Kepemimpinan
transformasional
memiliki visi dan kemampuan berinteraksi
dengan baik kepada karyawan, sehingga
dapat
memotivasi
karyawan
dalam
mewujudkan visi dan misi perusahaan. Gaya
kepemimpinan
transformasional
menekankan pada proses dimana orang yang
terlibat dengan orang lain akan menciptakan
suatu
hubungan
motivasi
dalam
diri
pemimpin dengan pengikut. Kepemimpinan
transformasional adalah pemimpin yang
menginspirasi para pengikutnya untuk
mengenyampingkan kepentingan pribadi
mereka demi kebaikan organisasi dan
mereka mampu memiliki pengaruh yang luar
biasa pada diri para pengikutnya. Mereka
menaruh perhatian terhadap kebutuhan
pengembangan
diri
para
pengikutnya, Rumusan Masalah Berdasarkan latar belakang masalah
serta betapa pentingnya kinerja pegawai
dalam pencapaian tujuan organisasi. Maka
di rumuskan rumusan masalah penelitian ini
adalah “Bagaimana pengaruh motivasi
intrinsik, komunikasi interpersonal dan
kepemimpinan transformasional terhadap
kepuasan kerja dalam meningkatkan kinerja
pegawai di Kantor Kementerian Agama
Kota
Serang.”
Selanjutnya
pertanyaan
penelitian yang diajukan dalam penelitian ini
adalah sebagai berikut: p
g
y
g
Selanjutnya menurut Satriowati at al. (2016) pada prinsipnya kepemimpinan
transformasional
memotivasi
karyawan
untuk berbuat lebih baik dari apa yang bisa
dilakukan,
dengan
kata
lain
dapat
meningkatkan keyakinan diri karyawan
yang nantinya akan berpengaruh terhadap
peningkatan kinerja karyawan. Keempat,
Faktor kepuasan kerja atau dalam arti yang
lebih khusus kepuasan pegawai dalam
bekerja memiliki peranan yang strategis di
dalam organisasi, bahkan dapat dikatakan
bahwa kepuasan kerja pegawai merupakan
salah satu kunci pendorong yang dapat
memengaruhi kinerja pegawai, hal tersebut
senada dengan yang diungkapkan oleh
Hasibuan (2010:203) bahwa kepuasan kerja
pegawai merupakan kunci pendorong moral
kerja, kedisiplinan dan prestasi kerja
pegawai dalam mendukung terwujudnya
tujuan
organisasi. Menurut
Robbins
(2010:139) istilah kepuasan kerja (job
satisfaction) merujuk pada sikap umum
seorang individu terhadap pekerjaannya. Selanjutnya
Siagian
(2011:295)
mengemukakan bahwa kepuasan kerja
merupakan suatu cara pandang seseorang
baik yang bersifat positif maupun negatif
tentang pekerjaannya. 1. Apakah terdapat pengaruh motivasi
intrinsik
terhadap
kepuasan
kerja
pegawai di Kantor Kementerian Agama
Kota Serang? 2. Apakah terdapat pengaruh komunikasi
interpersonal terhadap kepuasan kerja
pegawai di Kantor Kementerian Agama
Kota Serang? 3. Apakah
terdapat
pengaruh
kepemimpinan transformasional terhadap
kepuasan kerja pegawai di Kantor
Kementerian Agama Kota Serang? 4. Apakah terdapat pengaruh motivasi
intrinsik terhadap kinerja pegawai di
Kantor
Kementerian
Agama
Kota
Serang? 5. Apakah terdapat pengaruh komunikasi
interpersonal terhadap kinerja pegawai di
Kantor
Kementerian
Agama
Kota
Serang? 6. Apakah
terdapat
pengaruh
kepemimpinan transformasional terhadap
kinerja pegawai di Kantor Kementerian
Agama Kota Serang? Sejak diberlakukannya PP No. 6
tahun 2011, Kantor Kementerian Agama
Kota Serang telah melaksanakan penilain
prestasi kerja pegawai sebanyak tiga kali
yakni pada tahun 2014, tahun 2015 dan
tahun 2016, pada tahun 2016 menunjukkan
adanya
penurunan
jumlah
presentase
pegawai
yang
prestasi
kerjanya 7. Apakah terdapat pengaruh kepuasan kerja
pegawai terhadap kinerja pegawai di
Kantor
Kementerian
Agama
Kota
Serang? 46 46 JRBM Tirtayasa Volume 2 (1) - Juni 2018 interpersonal. Komunikasi
interpersonal
adalah penyampaian pesan oleh satu orang
dan penerimaan pesan oleh orang lain atau
sekelompok kecil orang, dengan berbagai
dampaknya dan dengan peluang untuk
memberikan umpan balik segera. Landasan Teori
Motivasi Intrinsik Menurut teori dikembangkan oleh
Herzberg yang dikenal dengan Two Factors
Theory menyatakan bahwa motivasi dibagi
menjadi dua faktor yaitu faktor motivator
atau yang sering disebut dengan motivasi
intrinsik dan faktor hygiene atau yang sering
disebut dengan motivasi ekstrinsik. Motivasi
intrinsik adalah motivasi yang datang dari
dalam diri seseorang sedangkan motivasi
ekstrinsik adalah motivasi yang disebabkan
oleh faktor eksternal atau faktor dari luar diri
seseorang. Motivasi
intrinsik
adalah
motivasi yang mendorong seseorang untuk
berprestasi yang bersumber dari dalam diri
individu tersebut. Efektivitas komunikasi interpersonal
dalam pandangan humanistik, komunikasi
dapat
berlangsung
efektif
dengan
memerhatikan empat indikasi, yaitu: 1)
Openness, yaitu meliputi aspek keinginan
untuk terbuka bagi masing-masing pihak
yang berinteraksi dan aspek keinginan untuk
menanggapi secara jujur segala stimulus
yang datang kepadanya. 2) Empathy, yaitu
merupakan kemampuan seseorang untuk
merasakan apa yang dirasakan oleh orang
lain, membayangkan apa yang dirasakan
orang lain dan turut terlibat secara emosional
dan intelektual dalam pengalaman orang
lain. 3) Positiveness, yaitu sikap positif yang
dikembangkan oleh individu-individu yang
berinteraksi. 4)
Equality, yaitu faktor
kesamaan yang dimiliki individu-individu
yang terlibat dalam suatu komunikasi. Faktor
kesamaan
ini
meliputi
nilai,
kepercayaan, pendidikan, status sosial dan
sebagainya. Empat
dimensi
tersebut
selanjutnya akan dijadikan sebagai indikator
dari variabel komunikasi interpersonal. Menurut Herzberg
yang tergolong
sebagai faktor motivator atau motivasi
intrinsik antara lain terdiri dari empat
indikasi, yaitu: 1)
Achievement, yaitu
keberhasilan
seorang
karyawan
dalam
menyelesaikan tugas. 2) Advancement, yaitu
suatu
keinginanan
seseorang
untuk
mengembangkan diri yang berhubungan
dengan pekerjaannya. 3) Work it self, yaitu
pekerjaan itu sendiri yang merupakan bentuk
dari variasi pekerjaan dan kontrol atas
metode serta langkah-langkah kerja. 4)
Recognition, yaitu pengakuan terhadap
karyawan dari organisasi atau perusahaan
atas prestasi kerja yang telah dicapai. Keempat dimensi tersebut selanjutnya akan
dijadikan sebagai indikator dari variabel
motivasi intrinsik. Komunikasi Interpersonal Kata komunikasi berasal dari Bahasa
latin communist atau common yang dalam
Bahasa Inggris berarti sama. Dengan
demikan berkomunikasi juga dapat diartikan
usaha untuk mencapai kesamaan makna atau
arti
commonness, atau dengan melalui
komunikasi mencoba berbagi informasi,
gagasan atau sikap kita dengan partisipan
lainnya. Proses
pertukaran
informasi
diantara seseorang dengan paling kurang
seorang lainnya atau biasanya di antara dua
orang
yang dapat langsung diketahui
balikannya
di
sebut
komunikasi 47 Jurnal Riset Bisnis dan Manajemen Tirtayasa seorang
individu
dengan
kebutuhan,
kemampuan, dan aspirasi yang berbeda,
serta melatih dan memberikan saran. Individualized
considerasion
dari
kepemimpinan
transformasional
memperlakukan masing-masing bawahan
sebagai individu serta mendampingi mereka,
memonitor dan menumbuhkan peluang. 5)
Idealized Behaviours (perilaku ideal), yaitu
perilaku
pemimpinyang
menekankan
pentingnya
konsekuensi
moral
dalam
memimpin. Lima
dimensi
tersebut
selanjutnya akan dijadikan sebagai indicator
dari
variabel
kepemimpinan
f
i
l mengubah kesadaran para pengikut atas isu-
isu yang ada dengan cara membantu orang
lain memandang masalah lama dengan cara
yang baru, serta mampu menyenangkan hati
dan menginspirasi para pengikutnya untuk
bekerja keras guna mencapai tujuan-tujuan
bersama. Kepemimpinan
transformasional
tercermin dari perilaku yang ditampilkan
oleh seorang pemimpin yang mengandung
lima indikasi yaitu : 1) Idealized influence
(pengaruh ideal), yaitu perilaku pemimpin
yang
memberikan
visi
danmisi,
memunculkan
rasa
bangga,
serta
mendapatkan
respek
(respect)
dan
kepercayaan bawahan (trust). Idealized
influence disebut juga sebagai pemimpin
yang
kharismatik,
dimana
pengikut
memiliki keyakinan yang mendalam pada
pemimpinnya, merasa bangga bisa bekerja
dengan
pemimpinnya,
danmempercayai
kapasitas pemimpinnya dalam mengatasi
setiap permasalahan. Idealized Influence
mengandung makna saling berbagi resiko,
melalui pertimbangan atas kebutuhan yang
dipimpin di atas kebutuhan pribadi, dan
perilaku moral serta etis. 2) Inspirasional
motivation
(motivasi
inspiratif),
yaitu
perilaku
pemimpin
yang
mampu
mengkomunikasikan harapan yang tinggi,
menyampaikan visi bersama secara menarik
dengan menggunakan simbol-simbol untuk
memfokuskan
upaya
bawahan,
dan
menginspirasi bawahan untuk mencapai
tujuan yang menghasilkan kemajuan penting
bagi organisasi. 3) Intellectual stimulation
(stimulasi
intelektual),
yaitu
perilaku
pemimpin yang mampu meningkatkan
kecerdasan bawahan untuk meningkatkan
kreativitas
dan
inovasi
mereka,
meningkatkan rasionalitas, dan pemecahan
masalah secara cermat, mendorong pengikut
untuk menanyakan asumsi, memunculkan
ide-ide dan metode-metode baru, dan
mengemukakan pendekatan lama dengan
cara
perspektif
baru. 4)
Individual
considerasion
(Pertimbangan
individu),
yaitu perilaku pemimpin yang memberikan
perhatian pribadi, memperlakukan masing-
masing bawahan secara individual sebagai Kinerja Kinerja berasal dari kata performance,
prestasi
kerja
atau
penampilan
kerja
(performance) diartikan sebagai ungkapan
kemampuan
yang
didasari
oleh
pengetahuan, sikap, dan ketrampilan dan
motivasi dalam menghasilkan sesuatu. Kinerja (prestasi kerja) adalah hasil kerja
secara kualitas dan kuantitas yang dicapai
oleh seorang pegawai dalam melaksanakan
tugasnya sesuai dengan tanggung jawab
yang diberikan kepadanya. Kinerja adalah
pencapaian yang optimal sesuai dengan
potensi yang dimiliki seorang karyawan
merupakan
hal
yang
selalu
menjadi
perhatian
para
pemimpin
organisasi. Sementara menurut peraturan pemerintah
(PP) nomor 46 tahun 2011 tentang penilaian
prestasi kerja pegawai, dalam PP tersebut
yang di maksud dengan prestasi kerja adalah
hasil kerja yang dicapai oleh setiap pegawai
pada satuan organisasi sesuai dengan sasaran
kerja pegawai dan perilaku kerja. Setidaknya
ada enam indikator atau kriteria untuk
mengukur kinerja pegawai secara individu,
yaitu: 1) Kualitas, hasil pekerjaan yang
dilakukan
mendekati
sempurna. 2)
Kuantitas, jumlah yang dihasilkan atau
jumlah aktivitas yang dapat diselesaikan. 3)
Ketepatan waktu, dapat menyelesaikan tepat
pada waktunya dan dapat memaksimalkan
waktu yang tersedia untuk kegiatan lain. 4)
Efektivitas, Pemanfaatan secara maksimal Dari beberapa indikator yang telah
diuraikan di atas yang akan diteliti oleh
penulis,
yang
selanjutnya
dijadikan
indikator dari variabel kinerja pegawai
adalah 1) Kualitas kerja, 2) kuantitas kerja,
3) ketepatan waktu, 4) biaya, dan 5) disiplin. JRBM Tirtayasa sumber daya yang ada pada organisasi untuk
meningkatkan keuntungan dan mengurangi
kerugian. 5)
Kemandirian,
dapat
melakasanakan kerja tanpa bantuan guna
menghindari hasil yang merugikan; 6)
Komitmen kerja, komitmen kerja antara
pegawai dengan organisasinya. Sedangkan
menurut Peraturan Pemerintah No. 46 tahun
2011 disebutkan bahwa tolak ukur prestasi
kerja terdiri dari unsur sasaran kinerja
pegawai atau disingkat SKP dan perilaku
kerja. SKP adalah rencana kerja dan target
yang akan dicapai oleh seorang pegawai
yang terdiri dari aspek; 1) kuantitas, 2)
kualitas, 3) waktu, dan 4) biaya. Sedangkan
perilaku kerja adalah setiap tingkah laku,
sikap atau tindakan yang dilakukan oleh
pegawai atau tidak melakukan sesuatu yang
seharusnya
dilakukan
sesuai
dengan
ketentuan peraturan perundang-undangan
yang terdiri dari aspek; 1) orientasi
pelayanan, 2) integritas, 3) komitmen, 4)
disiplin,
5)
kerja
sama,
dan
6)
kepemimpinan. kepuasan
kerja
berhubungan
dengan
variabel-variabel seperti, tingkat absensi,
umur,
tingkat
pekerjaan,
dan
ukuran
organisasi perusahaan. Jika indikasi-indikasi
tersebut
mengisyaratkan
positif
dapat
dikatakan bahwa kepuasan kerja karyawan
rendah, sebaliknya jika indikasi-indikasi
tersebut
mengisyaratkan
negatif
dapat
dikatakan bahwa kepuasan kerja karyawan
tinggi. Dari beberapa ahli tentang indikasi-
indikasi tersebut yang akan diteliti oleh
penulis,
yang
selanjutnya
dijadikan
indikator dari kepuasan kerja adalah 1)
prestasi kerja, 2) turnover atau perpindahan,
3) tingkat absensi, dan 4) Ukuran organisasi
atau perusahaan. Kepuasan Kerja Istilah
kepuasan
kerja
(job
satisfaction) merujuk pada sikap umum
seorang individu terhadap pekerjaannya. Kepuasan adalah suatu perasaan positif
tentang
pekerjaan
seseorang
yang
merupakan
hasil dari sebuah evaluasi
karakteristiknya. Kepuasan kerja adalah
keadaan emosi yang senang atau emosi
positif yang berasal dari penilaian pekerjaan
atau pengalaman kerja seseorang. Kepuasan
kerja
adalah
sikap
emosional
yang
menyenangkan dan mencintai pekerjaannya. Kepuasan kerja adalah pandangan pegawai
yang
menyenangkan
atau
tidak
menyenangkan terhadap pekerjaan mereka. Perasaan tersebut akan tampak dari sikap
positif karyawan terhadap pekerjaan dan
segala sesuatu yang dihadapi di lingkungan
kerjanya. Kepuasan kerja adalah suatu
perasaan yang menyokong atau tidak
menyokong diri pegawai yang berhubungan
dengan
pekerjaannya
maupun
dengan
kondisi dirinya. Kualitas kepuasan kerja dapat dilihat
dari indikasi yang ditampilkan melalui sikap
para karyawan terhadap pekerjaannya. indikasi kepuasan kerja hanya diukur dengan
prestasi kerja, kedisiplinan, moral kerja dan
turnover kecil maka secara relatif kepuasan
kerja karyawan baik. Sebaliknya jika
kedisiplinan, moral kerja, dan turnover
karyawan besar maka kepuasan
kerja
karyawan di perusahaan kurang8. Selain itu 48 48 Volume 2 (1) - Juni 2018 JRBM Tirtayasa Pengembangan Hipotesis
Pengaruh Motivasi Intrinsik terhadap
Kepuasan Kerja Menurut Nawawi (2010) motivasi
intrinsik adalah pendorong kerja yang
bersumber dari dalam diri pekerja sebagai
individu
berupa
kesadaran
mengenai
pentingnya atau manfaat atau makna
pekerjaan yang dilaksanakannya. Motivasi
yang ada pada diri seseorang merupakan
kekuatan pendorong yang akan mewujudkan
suatu perilaku guna mencapai tujuan
kepuasan dirinya. Rivai dan Bisri (2010)
mengatakan bahwa hubungan Motivasi
terhadap kepuasan kerja adalah apabila
individu termotivasi, mereka akan membuat
pilihan yang positif untuk melakukan
sesuatu, karena dapat memuaskan keinginan
mereka. 49 Jurnal Riset Bisnis dan Manajemen Tirtayasa Berbagai penelitian terdahulu yang
berkaitan
dengan
pengaruh
motivasi
intrinsik terhadap kepuasan kerja telah
dilakukan. Beberapa diantaranya penelitian
telah dilakukan oleh Akhtar at al. (2016),
Taufik. (2016),
Raza
at
al. (2016),
Cheraghalizadeh (2014), dan Budianto dan
Sambung (2013) yang menyatakan bahwa
motivasi intrinsik berpengaruh positif dan
signifikan terhadap kepuasan kerja pegawai. Oleh karena itu dalam penelitian ini diajukan
hipotesis pertama sebagai berikut: terhadap terhadap kepuasan kerja telah
dilakukan. Beberapa diantaranya telah
dilakukan oleh Putra. (2016), Jacobsat
(2016), Ibrahim (2016), Roebuck (2014),
dan Hakim at al. (2013) yang menyatakan
bahwa
komunikasi
interpersonal
berpengaruh positif dan signifikan terhadap
kepuasan kerja pegawai. Oleh karena itu
dalam penelitian ini diajukan hipotesis
kedua sebagai berikut: H2: Komunikasi interpersonaal berpengaruh
positif dan signifikan terhadap kepuasan
kerja. H2: Komunikasi interpersonaal berpengaruh
positif dan signifikan terhadap kepuasan
kerja. H1: Motivasi intrinsik berpengaruh positif
dan signifikan terhadap kepuasan kerja H1: Motivasi intrinsik berpengaruh positif
dan signifikan terhadap kepuasan kerja Pengaruh Motivasi Intrinsik terhadap
Kinerja Pegawai j
g
Menurut Nawawi (2010) motivasi
intrinsik adalah pendorong kerja yang
bersumber dari dalam diri pekerja sebagai
individu
berupa
kesadaran
mengenai
pentingnya atau manfaat atau makna
pekerjaan yang dilaksanakannya. Menurut
Hasibuan
(2010:143)
motivasi
adalah
pemberian
daya
penggerak
yang
menciptakan kegairahan kerja seseorang
agar mau bekerja sama, bekerja efektif, dan
terintegrasi dengan segala daya upayanya. Untuk mewujudkan kinerja individu yang
optimal, salah satunya adalah dengan
memberikan
motivasi
atau
dorongan
sehingga diharapkan akan tercapai sebuah
prestasi kerja. H5: Komunikasi interpersonal berpengaruh
positif dan signifikan terhadap kinerja
pegawai JRBM Tirtayasa Putra dan Ardana (2016), Nasra dan
Heilbrunn (2015), Long at al. (2014) dan
Braun at al. (2013) yang menyatakan bahwa
motivasi intrinsik berpengaruh positif dan
signifikan terhadap kepuasan kerja pegawai. Oleh karena itu dalam penelitian ini diajukan
hipotesis keenam sebagai berikut: kurang seorang lainnya atau biasanya di
antara dua orang yang dapat langsung
diketahui balikannya. Lebih lanjut Handoko
(2010:272) menjelaskan bahwa komunikasi
merupakan proses memberikan pemahaman
berupa ide atau informasi kepada seseorang,
komunikasi yang baik akan membantu
karyawan dalam memahami pekerjaannya
secara jelas sehingga perlahan kinerjanya
akan meningkat. H3: Kepemimpinan transformasional
berpengaruh positif dan signifikan
terhadap kepuasan kerja. Berbagai penelitian terdahulu yang
berkaitan dengan
pengaruh komunikasi
terhadap kinerja pegawai telah dilakukan. Beberapa daiantaranya telah dilakukan oleh
Atambo dan Momanyi (2016), Marjianto
(2015), Utami (2015), Femi (2014) dan
Usman (2013) yang menyatakan bahwa
komunikasi
interpersonal
berpengaruh
positif dan signifikan terhadap kinerja
pegawai. Oleh karena itu dalam penelitian
ini diajukan hipotesis kelima sebagai
berikut: Pengaruh Kepemimpinan
Transformasional terhadap Kinerja
Pegawai g
Menurut Hughes et al. (2012:542)
bahwa pemimpin transformasional memiliki
visi dan kemampuan berinteraksi dengan
baik kepada karyawan, sehingga dapat
memotivasi karyawan dalam mewujudkan
visi dan misi perusahaan. Sedangkan
menurut Yukl (2010) menyebutkan bahwa
dengan kepemimpinan transformasional,
para pegawai merasakan kepercayaan,
kekaguman, kesetiaan dan penghormatan
terhadap pemimpin dan para pegawai
termotivasi untuk melakukan pekerjaan
melampaui yang diharapkan sebelumnya. Lebih lanjut Yukl (2010) menjelaskan
bahwa
kepemimpinan
transformasional
akan mengarahkan pemimpin meningkatkan
motivasi, moral dan kinerja para pegawai
melalui berbagai mekanisme, termasuk
menghubungkan
pengertian
pegawai
tentang identitas diri untuk misi dan identitas Berbagai penelitian terdahulu yang
berkaitan
dengan
pengaruh
motivasi
terhadap kinerja telah dilakukan. Beberapa
diantaranya telah dilakukan oleh Linawati
(2014), Dirianzani at al (2014), Budianto
dan Sambung (2013), Dhermawan at al
(2012) dan Muslih (2012) yang menyatakan
bahwa motivasi intrinsik tidak berpengaruh
signifikan terhadap kinerja pegawai. Oleh
karena itu dalam penelitian ini diajukan
hipotesis keempat sebagai berikut: H4: Motivasi intrinsik berpengaruh positif
dan signifikan terhadap kinerja pegawai H4: Motivasi intrinsik berpengaruh positif
dan signifikan terhadap kinerja pegawai Pengaruh Komunikasi Interpersonal
terhadap Kepuasan Kerja j
Menurut
Robbins
dan
Judge
(2015:90)
pemimpin
transformasional
adalah pemimpin yang menginspirasi para
pengikutnya
untuk
mengenyampingkan
kepentingan pribadi mereka demi kebaikan
organisasi dan mereka mampu memiliki
pengaruh yang luar biasa pada diri para
pengikutnya. Mereka menaruh perhatian
terhadap kebutuhan pengembangan diri para
pengikutnya, mengubah kesadaran para
pengikut atas isu-isu yang ada dengan cara
membantu orang lain memandang masalah
lama dengan cara yang baru, serta mampu
menyenangkan hati dan menginspirasi para
pengikutnya untuk bekerja keras guna
mencapai tujuan-tujuan bersama. Handoko (2010:272) menjelaskan
bahwa
komunikasi
merupakan
proses
memberikan pemahaman berupa ide atau
informasi kepada seseorang. Komunikasi
yang baik akan membantu karyawan dalam
memahami
pekerjaannya
secara
jelas
sehingga
perlahan
kinerjanya
akan
meningkat. Penyampaian pesan oleh satu
orang dan penerimaan pesan oleh orang lain
atau sekelompok kecil orang, dengan
berbagai dampaknya dan dengan peluang
untuk memberikan umpan balik segera di
sebut komunikasi interpersonal (Devito
dalam Effendy, 2010:23). Komunikasi interpersonal menjadi
salah satu yang diharapkan oleh pegawai
untuk meminimalisasi masalah individu dan
mampu menciptakan hubungan yang akrab
antar rekan kerja, yang akhirnya dapat
membantu dalam pemenuhan kebutuhan
social sehingga dapat mencapai tingkat
kepuasan kerja dan kinerja yang diharapkan
(Putr, 2016). Lebih lanjut menurut Arni
(2011:85) mengemukakan bahwa untuk
mengatasi rasa ketidakpuasan kerja dapat
dilakukan dengan memberikan informasi
yang cukup kepada karyawan sehingga
mereka dapat melakukan pekerjaan dengan
baik dan merasa puas dengan hasil yang
dilakukannya. Karakteristik
pemimpin
transformasional
adalah
kharismatik,
inspiratif,
mampu
memotivasi,
dan
menghargai perbedaan kemampuan individu
(Robin dan Judge, 2015). Pemimpin yang
selalu hadir di tengah-tengah karyawan,
memberi suri tauladan tentang komitmen
kerja,
menginspirasi
dan
memotivasi
karyawan, memberi penghargaan yang adil,
tentu akan membuat karyawan merasa puas
(Thamrin, 2012). Berbagai penelitian yang berkaitan
dengan
pengaruh
kepemimpinan
transformasonal terhadap kepuasan kerja
telah dilakukan. Beberapa diantaranya telah
dilakukan oleh Muhammad at al. (2016), Berbagai penelitian terdahulu yang
berkaitan dengan pengaruh komunikasi 50 50 Volume 2 (1) - Juni 2018 JRBM Tirtayasa Pengaruh Komunikasi Interpersonal
terhadap Kinerja Pegawai Komunikasi interpersonal menurut
Arni (2011) adalah proses pertukaran
informasi diantara seseorang dengan paling 51 Jurnal Riset Bisnis dan Manajemen Tirtayasa kolektif organisasi, menjadi teladan bagi
pegawai yang mengilhami pegawai yang
bersangkutan, menantang pegawai untuk
mengambil kepemilikan yang lebih besar
untuk
pekerjaannya,
dan
memahami
kekuatan dan kelemahan dari pegawai,
sehingga pemimpin dapat menyesuaikan
pegawai
dengan
tugas-tugas
yang
mengoptimalkan kinerjanya. kolektif organisasi, menjadi teladan bagi
pegawai yang mengilhami pegawai yang
bersangkutan, menantang pegawai untuk
mengambil kepemilikan yang lebih besar
untuk
pekerjaannya,
dan
memahami
kekuatan dan kelemahan dari pegawai,
sehingga pemimpin dapat menyesuaikan
pegawai
dengan
tugas-tugas
yang
mengoptimalkan kinerjanya. pegawai, hal tersebut senada dengan yang
diungkapkan oleh Hasibuan (2010:203)
bahwa kepuasan kerja pegawai merupakan
kunci pendorong moral kerja, kedisiplinan
dan
prestasi
kerja
pegawai
dalam
mendukung terwujudnya tujuan organisasi. Berbagai penelitian terdahulu yang
berkaitan dengan pengaruh kepuasan kerja
terhadap terhadap kinerja telah dilakukan. Beberapa diantaranya telah dilakukan oleh
Babalola (2016), Siengthai dan Pila-Ngarm
(2016), Putra (2016), Taufik (2016) dan
Javed at al.(2014) yang menyatakan bahwa
kepuasan kerja berpengaruh positif dan
signifikan terhadap kinerja pegawai. Oleh
karena itu dalam penelitian ini diajukan
hipotesis ketujuh sebagai berikut: Berbagai penelitian terdahulu yang
berkaitan dengan pengaruh kepemimpinan
transformasional terhadap kinerja pegawai
telah dilakukan. Beberapa diantaranya telah
dilakukan oleh Ariyabuddhiphong dan Kahn
(2017), Yoshi Tania (2017), Arthawan dan
Mujiati (2017), Masa’deh at al.(2016) dan
Fernadi at al. (2016) yang menyatakan
bahwa
kepemimpinan
transformasional
berpengaruh positif dan signifikan terhadap
kinerja pegawai. Oleh karena itu dalam
penelitian ini diajukan hipotesis keenam
sebagai berikut: H7: Kepuasan kerja berpengaruh positif dan
signifikan terhadap kinerja pegawai Berdasarkan hasil tinjaun pustaka
dan analisis terhadap bukti-bukti empiris
dari penelitian terdahulu maka variabel yang
akan diteliti dalam penelitian ini adalah
motivasi
intrinsik,
komunikasi
interpersonal,
kepemimpinan H6: Kepemimpinan transformational
berpengaruh positif dan signifikan
terhadap kinerja pegawai. H6: Kepemimpinan transformational
berpengaruh positif dan signifikan
terhadap kinerja pegawai. Pengaruh Kepuasan Kerja terhadap
Kinerja Pegawai transfprmasional terhadap kinerja pegawai,
dengan menambahkan variabel kepuasan
kerja sebagai variabel intervening. Dari
variabel-variabel tersebut, model yang
dikembangkan
untuk
diteliti
dalam
penelitian ini disajikan dalam gambar
berikut: Faktor kepuasan kerja memiliki
peranan yang strategis di dalam organisasi,
bahkan dapat dikatakan bahwa kepuasan
kerja pegawai merupakan salah satu kunci
pendorong yang dapat memengaruhi kinerja Gambar 1 Model Penelitian Gambar 1 Model Penelitian Gambar 1 Model Penelitian 52 52 JRBM Tirtayasa Volume 2 (1) - Juni 2018 bersifat
soft
modeling
karena
tidak
mengasumsikan
data
harus
dengan
pengukuran skala tertentu, yang berarti
jumlah sampel dapat kecil (dibawah 100
sampel)23. Hasil Pengujian Kualitas Data (Outer
Model) Terdapat tiga kriteria di dalam
penggunaan teknik analisa data dengan
SmartPLS untuk menilai outer model yaitu
Convergent Validity, Discriminant Validity,
Cronbach Alfa dan Composite Reliability
(Chin, 1998 dalam Ghozali, 2011). Analisis Partial Least Squares (PLS)
adalah teknik statistika multivarian yang
melakukan perbandingan antara variabel
dependen berganda dan variabel independen
berganda. PLS merupakan salah satu metode
statistika SEM berbasis varian yang didesain
untuk menyelesaikan regresi berganda
ketika terjadi permasalahan spesifik pada
data22. PLS adalah metode analisis yang Metode Penelitian Dalam penelitian ini digunakan
teknik
analisis
Structural
Equation
Modelling (SEM) yang merupakan analisis
multivariate. SEM adalah keluarga model
statistik yang mencari untuk menjelaskan
hubungan antara variabel yang banyak. SEM
adalah statistik yang unik karena fondasinya
berada di antara dua tehnik multivariate
yang familiar yaitu faktor analisis dan
analisa regresi berganda21. Selanjutnya
untuk melakukan analisis SEM diperlukan
tools atau software dalam penelitian ini
menggunakan SmartPLS. ANALISIS DATA Hasil Pengujian Kualitas Data (Outer
Model) Convergent Validity Convergent validity dari model
pengukuran
dengan
refleksif
indikator
dinilai berdasarkan korelasi antara item
score/component score yang diestimasi
dengan Soflware PLS. Ukuran refleksif
individual dikatakan tinggi jika berkorelasi
lebih dari 0,70 dengan konstruk yang diukur. Gambar 2 Output pengukuran model Gambar 2 Output pengukuran model Berdasarkan gambar 3 pengukuran
model
kedua,
setelah
menghilangkan
loading factor yang rendah, model terlihat
sudah memiliki convergen validity karena
tidak terdapat lagi loading factor yang
nilainya di bawah 0.7, sehingga model sudah
layak untuk dianalisis lebih lanjut. Berdasarkan
gambar
2
output
pengukuran model terlihat bahwa masih
terdapat hasil outer loading yang nilainya di
bawah 0.70 yaitu MI.3 dan KI.2, karena
tidak memenuhi sarat yaitu nilainya kurang
dari
0.70,
maka
indikator
tersebut
dihilangkan, kemudian model di run
kembali, dan hasil pengukuran model kedua
terlihat pada gambar 3. 53 53 Jurnal Riset Bisnis dan Manajemen Tirtayasa Gambar 3 Output pengukuran model kedua Gambar 3 Output pengukuran model kedua Discriminant Validity Berdasarkan tabel 1 cross loading
dalam
pengujian
model
pengukuran
menunjukkan bahwa masih terdapat nilai
loading dari masing-masing item terhadap
konstruknya tidak lebih
besar dari pada
nilai cross loading-nya yaitu pada indikator
KK.4 dan KP.5, maka selanjutnya indikator
tersebut dihilangkan, kemudian model di run
kembali. Croos loading kedua terlihat pada
tabel 2. Cara
untuk
menguji
validitas
discriminant dengan indikator refleksif yaitu
dengan melihat nilai cross loading untuk
setiap variabel harus lebih besar dari 0.70,
dan nilai loading dari setiap indikator dari
sebuah variabel memiliki nilai loading
paling besar dengan nilai loading lain
terhadap variabel laten lainnya. Nilai
loading tersebut dapat dilihat pada tabel
cross loading pada menu descriminant
validity. Berdasarkan tabel 2 cross loading
kedua dalam pengujian model pengukuran di
atas menunjukkan bahwa semua nilai
loading dari masing-masing item terhadap
konstruknya lebih besar dari pada nilai
cross loading-nya, hal ini berarti bahwa
setiap variabel laten memiliki discriminant
validity yang baik dan tidak terdapat
permasalahan discriminant validity. Tabel 1 Output cross Loading Tabel 1 Output cross Loading 54 Volume 2 (1) - Juni 2018 Volume 2 (1) - Juni 2018 JRBM Tirtayasa JRBM Tirtayasa
Volume 2 (1)
Juni 2
Tabel 2 Output cross Loading kedua
Berikut adalah model penguk
ketiga setelah dipastikan bahwa model s
memiliki
convergent
validity
discriminant validity yang baik. Gambar 4 Output pengukuran model ketiga Tabel 2 Output cross Loading kedua Tabel 2 Output cross Loading kedua Berikut adalah model pengukuran
ketiga setelah dipastikan bahwa model sudah
memiliki
convergent
validity
dan
discriminant validity yang baik. Gambar 4 Output pengukuran model ketiga Gambar 4 Output pengukuran model ketiga Cronbach alpha dan Composite reliability Cronbach alpha dan Composite reliability Cronbach alpha dan Composite reliability Tabel 3 Cronbach alpha dan composite
reliability Tabel 3 Cronbach alpha dan composite
reliability
B d
k
b l 3 C
b
h l h Tabel 3 Cronbach alpha dan composite
reliability Untuk memastikan bahwa tidak ada
masalah terkait pengukuran, maka langkah
terakhir dalam evaluasi Outer model adalah
menguji unidimensionalitas dari model. Uji
unidimensionality
dilakukan
dengan
menggunakan indikator cronbach alpha dan
composite reliability. Untuk nilai kedua
indikator ini harus lebih besar dari 0.7022. Cronbach alpha dan Composite reliability
terdapat pada menu Construct Reliability
and validity. Berdasarkan tabel 3 Cronbach alpha
dan composite reliability menunjukan dari
masing-masing konstruk nilainya di atas 0.7 55 Jurnal Riset Bisnis dan Manajemen Tirtayasa sehingga dapat dinyatakan bahwa semua
konstruk yang dipakai dalam penelitian ini
memenuhi kriteria reliabel. merupakan kontribusi dari variabel motivasi
intriksik,
komunikasi
interpersonal,
kepemimpinan
transformasional
dan
kepuasan
kerja,
sedangkan
sisanya
merupakan kontribusi dari variabel lain
diluar model yang diajukan. Hasil Pengujian Kelayakan Model
(Inner Model) Pengujian
kelayakan
model
dilakukan dengan uji inner model atau
model struktural yang dilakukan untuk
melihat hubungan antara konstruk, nilai
signifikansi dan R-square dari model
penelitian. Nilai R-square digunakan untuk
mengukur tingkat variasi perubahan variabel
independen terhadap variabel dependen. Semakin tinggi nilai R-square berarti
semakin baik model prediksi dari model
penelitian
yang
diajukan
(Jogiyanto,
2011:72). Pada penelitian ini terdapat dua
variabel yang dipengaruhi oleh variabel
lainnya yaitu variabel kepuasan kerja dan
variabel kinerja pegawai. Dengan demikian
nilai R-Square dalam penelitian ini terdapat
pada variabel kepuasan kerja dan kinerja
pegawai. Untuk melihat nilai R-square
dalam PLS terdapat pada menu R-square
pada PLS Algorithm. Hasil Pengujian Hipotesis Hasil Pengujian Hipotesis
Menurut
Jogiyanto
(2011:86)
Ukuran
signifikansi
keterdukungan
hipotesis dapat digunakan perbandingan
nilai T-table dan T-statistic. Jika T-statistic
lebih tinggi dibandingkan nilai T-table,
berarti hipotesis terdukung atau diterima. Untuk tingkat keyakinan 95 persen (alpha
5%), maka nilai T-table adalah > 1,96. Selain
itu,
ukuran
signifikansi
keterdukungan hipotesis dapat dilihat dari
nilai p value, jika nilai P value lebih kecil
dari 0.05 maka hipotesis terdukung atau
diterima untuk tingkat keyakinan 95 persen
(alpha 5%). Untuk melihat nilai T-statistic dan P
Valuae dalam SmartPLS terdapat pada path
coefficient setelah dilakukan melalui proses
bootstraping terhadapa model yang sudah
valid dan reliabel serta memenuhi kelayakan
model. Tabel 4 R-Square Tabel 5 Path Coefficients Tabel 5 Path Coefficients Tabel 5 Path Coefficients Berdasarkan tabel 4 R-square untuk
variabel kepuasan kerja diperoleh sebesar
0.686, hal ini menunjukkan bahwa variasi
perubahan dari variabel kepuasan kerja
sebsar 68.6% merupakan kontribusi dari
variabel motivasi intriksik, komunikasi
interpersonal
dan
kepemimpinan
transformasional,
sedangkan
sisanya
merupakan kontribusi dari variabel lain
diluar model yang diajukan. Kemudian nilai
R-Square untuk variabel kinerja pegawai
diperoleh sebesar 0.782, hal ini menunjukan
bahwa variasi perubahan dari variabel
variabel kinerja pegawai sebesar 78.2% Berdasarkan tabel 5 Path coeffisient,
dilihat dari nilai koefisien parameter
(original sample) semua menunjukan nilai
yang positif, hal ini berarti pengaruh antar
variabel menunjukan pengaruh yang positif. Sedangkan dilihat dari uji signifikansi dari
tujuh hipotesis yang diajukan terdapat enam
hipotesis yang terdukung atau diterima
karena nilai T-Statistic-nya lebih besar dari
1.96 dan P Value lebih kecil dari 0.05, yaitu
motivasi intrinsik berpengaruh positif dan
signifikat
terhadap
kepuasan
kerja,
komunikasi
interpersonal
berpengaruh 56 Volume 2 (1) - Juni 2018 JRBM Tirtayasa dependen (kinerja pegawai). Hasil uji
signifikansi menunjukan bahwa variabel
kepuasan kerja memidiasi penuh (fully
mediating) pengaruh variabel motivasi
intrinsik terhadap variabel kinerja pegawai,
variabel komunikasi interpersonal terhadap
variabel kinerja pegawai, dan variabel
kepemimpinan transformasional terhadap
variabel kinerja pegawai, hal tersebut
ditunjukan oleh nilai T-Statistik yang lebih
besar di bandingkan dengan T-Table (1.960)
dan nilai P Value lebih kecil dari 0.05 pada
alpha 5%. positif dan signifikan terhadap kepuasan
kerja,
kepemimpinan
transformasional
berpengaruh positif dan signifikan terhadap
kepuasan
kerja,
motivasi
intrinsik
berpengaruh positif dan signifikan terhadap
kinerja
pegawai,
kepemimpinan
transformasional berpengaruh positif dan
signifikan
terhadap
terhadap
kinerja
pegawai, dan kepuasan kerja berpengaruh
positif dan signifikan terhadap kinerja
pegawai. Hasil Pengujian Hipotesis Sedangkan satu hipotesis sisanya
tidak terdukung atau ditolak karena nilai T-
Statistic-nya lebih kecil dari 1.96 dan P
Value
lebih
besar
dari
0.05,
yaitu
komunikasi
interpersonal
berpengaruh
positif namun tidak signifikan terhadap
kinerja pegawai.. Hasil dan Pembahasan
Pengaruh Motivasi Intrinsik terhadap
Kepuasan Kerja Hasil
pengujian
hipotesis
menunjukan
bahwa
motivasi
intrinsik
berpengaruh positif terhadap kepuasan kerja,
hal ini dapat dilihat dari koefisien parameter
(original sample) sebesar 0.199 (positif),
sedangkan hasil uji signifikansi menunjukan
nilai T-Statistic sebesar 2.589 lebih besar
dari T-Table (2.589>1.960) dan nilai P value
sebesar 0.005 lebih kecil dari dari 0.05 pada
alpha 5%, hal ini menunjukan hipotesis
terdukung
atau
diterima
(signifikan). Dengan demikian, dapat disimpulkan bahwa
motivasi intrinsik berpengaruh positif dan
signifikan terhadap kepuasan kerja. Hasil Pengujian Efek Mediasi Efek mediasi menunjukan hubungan
antara variabel independen dan dependen
melalui variabel penghubung atau mediasi. Efek mediasi diuji ketika diduga terdapat
variabel
intervening
diantara
variabel
independen
dan
dependen. Artinya,
pengaruh variabel independen terhadap
variabel dependen tidak secara langsung
terjadi tetapi melalui proses transformasi
yang
diwakili
oleh
variabel22. Pada
pengujian efek mediasi, ouput parameter uji
signifikansi dapat dilihat pada total indirect
effects. Hasil pengujian model dengan efek
moderasi pada penelitian ini dapat dilihat
pada tabel indirect effects. Hasil penelitian tersebut sejalan
dengan penelitian yang telah dilakukan oleh
Akhtar at al. (2016), Taufik. (2016), Raza at
al. (2016), Cheraghalizadeh (2014) dan
Budianto dan Sambung
(2013)
yang
menyatakan
bahwa
motivasi
intrinsik
berpengaruh positif dan signifikan terhadap
kepuasan kerja pegawai. Tabel 6 Indirect effects Pengaruh Komunikasi Interpersonal
terhadap Kinerja Pegawai
Hasil
pengujian
hipot Hasil
pengujian
hipotesis
menunjukan
komunikasi
interpersonal
berpengaruh
positif
terhadap
kinerja
pegawai, hal ini dapat dilihat dari koefisien
parameter (original sample) sebesar 0.012
(positif), sedangkan hasil uji signifikansi
menunjukan nilai T-Statistic sebesar 0.189
lebih kecil dari T-Table (0.189<1.960) dan
nilai P value sebesar 0.425 lebih besar dari
dari 0.05 pada alpha 5%, hal ini menunjukan
hipotesis tidak terdukung atau ditolak (tidak
signifikan). Dengan
demikian,
dapat
disimpulkan
bahwa
komunikasi
interpersonal berpengaruh positif namun
tidak signifikan terhadap kinerja pegawai. Hasil penelitian tersebut sejalan
dengan penelitian yang telah dilakukan oleh
Muhammad at al. (2016), Putra dan Ardana
(2016), Nasra dan Heilbrunn (2015), Long at
al. (2014) dan Braun at al. (2013) yang
menyatakan
bahwa
motivasi
intrinsik
berpengaruh positif dan signifikan terhadap
kepuasan kerja pegawai. Hasil penelitian tersebut berbeda
hasilnya dengan penelitian yang telah
dilakukan oleh Atambo dan Momanyi
(2016), Marjianto (2015), Utami (2015),
Femi (2014) dan Usman (2013) yang
menyatakan
bahwa
komunikasi Pengaruh Komunikasi Interpersonal
terhadap Kepuasan Kerja
Hasil
pengujian
hipot
menunjukan
bahwa
komuni Hasil
pengujian
hipotesis
menunjukan
bahwa
komunikasi
interpersonal berpengaruh positif terhadap
kepuasan kerja, hal ini dapat dilihat dari
koefisien
parameter
(original
sample)
sebesar 0.162 (positif), sedangkan hasil uji
signifikansi menunjukan nilai T-Statistic
sebesar 2.284 lebih besar dari T-Table Berdasarkan tabel 6 Indirect effects, Berdasarkan tabel 6 Indirect effects,
terlihat semua nilai parameter original
sampel menunjukan angka yang positif,
dengan demikian variabel kepuasan kerja
memdiasi secara positif pengaruh variabel
independen (motivasi intrinsik, komunikasi
interpersonal,
dan
kepemimpinan
transformasional)
terhadap
variabel 57 Jurnal Riset Bisnis dan Manajemen Tirtayasa (2.284>1.960) dan nilai P value sebesar
0.011 lebih kecil dari dari 0.05 pada alpha
5%, hal ini menunjukan hipotesis terdukung
atau
diterima
(signifikan). Dengan
demikian,
dapat
disimpulkan
bahwa
komunikasi
interpersonal
berpengaruh
positif dan signifikan terhadap kepuasan
kerja. (original sample) sebesar 0.250 (positif),
sedangkan sedangkan hasil uji signifikansi
menunjukan nilai T-Statistic sebesar 2.813
lebih besar dari T-Table (2.813>1.96) dan
nilai P value sebesar 0,003 lebih besar dari
0,05 pada alpha 5% menunjukan hipotesis
diterima. Dengan
demikian
dapat
disimpulkan
bahwa
motivasi
intrinsik
berpengaruh positif dan signifikan terhadap
kinerja pegawai. Hasil penelitian tersebut sejalan
dengan penelitian yang telah dilakukan oleh
Putra. (2016), Jacobsat (2016), Ibrahim
(2016), Roebuck (2014), dan Hakim at al. (2013) yang menyatakan bahwa komunikasi
interpersonal
berpengaruh
positif
dan
signifikan terhadap kepuasan kerja pegawai. Hasil penelitian tersebut sejalan
dengan penelitian yang telah dilakukan oleh
Wawuru (2017), Taufik (2016), Fernadi at
al. (2016), Lukito at al. (2016), dan EK dan
Makaru (2015) menunjukan bahwa motivasi
intrinsik berpengaruh positif dan signifikan
terhadap kinerja pegawai. Namun hasil
penelitian tersebut bertolak belakang dengan
hasil penelitian yang telah dilakukan oleh
Linawati (2014), Dirianzani at al (2014),
Budianto dan Sambung (2013), Dhermawan
at al (2012) dan Muslih (2012) yang
menyatakan bahwa motivasi intrinsik tidak
berpengaruh signifikan terhadap kinerja
pegawai. Pengaruh Kepemimpinan
Transformasional terhadap Kepuasan
Kerja j
Hasil
pengujian
hipotesis
menunjukan
bahwa
kepemimpinan
transformasional
berpengaruh
positif
terhadap kepuasan kerja, hal ini dapat dilihat
dari koefisien parameter (original sample)
sebesar 0.636 (positif), sedangkan hasil uji
signifikansi menunjukan nilai T-Statistic
sebesar 9.527 lebih besar dari T-Table
(9.527>1.960) dan nilai P value sebesar
0.000 lebih kecil dari dari 0.05 pada alpha
5%, hal ini menunjukan hipotesis terdukung
atau
diterima
(signifikan). Dengan
demikian,
dapat
disimpulkan
bahwa
kepemimpinan
transformasional
berpengaruh positif dan signifikan terhadap
kepuasan kerja. Pengaruh Motivasi Intrinsik terhadap
Kinerja Pegawai Hasil
pengujian
hipotesis
menunjukan motivasi intrinsik berpengaruh
positif terhadap kinerja pegawai, hal ini
dapat dilihat dari koefisien parameter 58 58 Volume 2 (1) - Juni 2018 JRBM Tirtayasa pegawai, hal ini dapat dilihat dari koefisien
parameter (original sample) sebesar 0.378
(positif), sedangkan hasil uji signifikansi
menunjukan nilai T-Statistic sebesar 3.905
lebih besar dari T-Table (3.905>1.960) dan
nilai P value sebesar 0.000 lebih kecil dari
dari 0.05 pada alpha 5%, hal ini menunjukan
hipotesis
terdukung
atau
diterima
(signifikan). Dengan
demikian,
dapat
disimpulkan
bahwa
kepuasan
kerja
berpengaruh positif dan signifikan terhadap
kinerja pegawai. interpersonal
berpengaruh
positif
dan
signifikan terhadap kinerja pegawai. Dilihat dari bebrapa tanggapan
responden komunikasi interpersonal yang
terjalin antar pegawai belum berjalan efektif
hal tersebut dikarenakan anatar pegawai
kurang adanya keterbukaan dalam hal
penyamapai informasi, terkadang hanya
orang-orang tertentu saja yang mendapatkan
informasi tersebut, sehingga menimbulkan
prasangka-prasangka yang tidak baik antar
pegawai. Selain itu, terkadang ketika ada
pegawai
yang
mendapatkan
masalah
terutama dalam hal pekerjaan, pegawai
lainnya kurang peduli dan kurang memiliki
rasa empati yang baik. Hasil penelitian tersebut sejalan
dengan penelitian yang telah dilakukan oleh
Babalola (2016), Siengthai dan Pila-Ngarm
(2016), Putra (2016), Taufik (2016) dan
Javed at al.(2014) yang menyatakan bahwa
kepuasan kerja berpengaruh positif dan
signifikan terhadap kinerja pegawai. pengujian
hipotesis
kepemimpinan Hasil
pengujian
hipotesis
menunjukan
kepemimpinan
transformasional
berpengaruh
positif
terhadap kinerja pegawai, hal ini dapat
dilihat dari koefisien parameter (original
sample) sebesar 0.363 (positif), sedangkan
hasil uji signifikansi menunjukan nilai T-
Statistic sebesar 3.753 lebih besar dari T-
Table (3.753>1.960) dan nilai P value
sebesar 0.000 lebih kecil dari dari 0.05 pada
alpha 5%, hal ini menunjukan hipotesis
terdukung
atau
diterima
(signifikan). Dengan demikian, dapat disimpulkan bahwa
kepemimpinan
transformasional
berpengaruh positif dan signifikan terhadap
kinerja pegawai. pengujian
hipotesis
kepemimpinan Dalam penelitian ini mengkaji lima
variabel, yaitu tiga variabel independen
(motivasi
intrinsik,
komunikasi
interpersonal
dan
kepemimpinan
transformational) satu variabel dependen
(kinerja
pegawai),
dan
satu
variabel
intervining (kepuasan kerja). Berdasarkanperhitungan
nilai
R-
square untuk variabel kepuasan kerja
diperoleh
sebesar
0.686,
hal
ini
menunjukkan bahwa variasi perubahan dari
variabel kepuasan kerja sebsar 68.6%
merupakan kontribusi dari variabel motivasi
intriksik, komunikasi interpersonal dan
kepemimpinan transformasional, sedangkan
sisanya merupakan kontribusi dari variabel
lain diluar model yang diajukan. Kemudian
nilai R-Square
untuk variabel kinerja
pegawai diperoleh sebesar 0.782, hal ini
menunjukan bahwa variasi perubahan dari
variabel variabel kinerja pegawai sebesar
78.2% merupakan kontribusi dari variabel
motivasi
intriksik,
komunikasi
interpersonal,
kepemimpinan Hasil penelitian tersebut sejalan
dengan penelitian yang telah dilakukan oleh
Ariyabuddhiphong dan Kahn (2017), Yoshi
Tania (2017), Arthawan dan Mujiati (2017),
Masa’deh at al.(2016), Fernadi at al. (2016)
yang menyatakan bahwa kepemimpinan
transformasional berpengaruh positif dan
signifikan terhadap kinerja pegawai. interpersonal, Jurnal Riset Bisnis dan Manajemen Tirtayasa terdukung
atau
diterima,
artinya
semakin
tinggi
penerapan
kepemimpinan transformasional maka
kinerja
pegawai
akan
semakin
meningkat. terdukung
atau
diterima,
artinya
semakin
tinggi
penerapan
kepemimpinan transformasional maka
kinerja
pegawai
akan
semakin
meningkat. Berdasarkan
hasil
pengujian
terhadap hipotesis yang diajukan diperoleh
kesimpulan sebagai berikut: 1. Hipotesis pertama yang menyatakan
bahwa motivasi intrinsik berpengaruh
positif dan signifikat terhadap kepuasan
kerja terdukung atau diterima, artinya
semakin tinggi motivasi intrinsik yang
dimiliki oleh pegawai maka kepuasan
kerja pegawai akan semakin meningkat. 7. Hipotesis ketujuh yang menyatakan
bahwa kepuasan kerja berpengaruh
positif
dan
signifikan
terhadap
kinerja pegawai terdukung atau
diterima, artinya semakin tinggi
tingkat kepuasan kerja pegawai maka
kinerja
pegawai
akan
semakin
meningkat. 2. Hipotesis kedua yang menyatakan
bahwa
komunikasi
interpersonal
berpengaruh positif dan signifikan
terhadap kepuasan kerja terdukung atau
diterima,
artinya
semakin
tingggi
efektivitas komunikasi interpersonal
(antar
personal
pegawai)
maka
kepuasan kerja pegawai akan semakin
meningkat. Pengaruh Kepuasan Kerja terhadap
kinerja pegawai transformasional
dan
kepuasan
kerja,
sedangkan sisanya merupakan kontribusi
dari variabel lain diluar model yang
diajukan. Hasil
pengujian
hipotesis
menunjukan
bahwa
kepuasan
kerja
berpengaruh
positif
terhadap
kinerja 59 Jurnal Riset Bisnis dan Manajemen Tirtayasa Saran Ilmiah Bagi peneliti yang akan melakukan
penelitian lebih lanjut dari penelitian ini
supaya menggunakan populasi sasaran lebih
luas agar diperoleh temuan yang lebih
representatif
tentang
kondisi
empirik
permasalahan sumber daya manusia
khususnya yang berkaitan dengan variabel
motivasi intrinsik, komunikasi interpersonal
dan kepemimpinan transformasional dalam
menyelesaikan
permasalahan
kinerja
pegawai. 3. Hipotesis ketiga yang menyatakan
bahwa kepemimpinan transformasional
berpengaruh positif dan signifikan
terhadap kepuasan kerja terdukung atau
diterima,
artinya
semakin
tinggi
penerapan
kepemimpinan
transformasional maka kepuasan kerja
pegawai
(bawahan)
akan
semakin
meningkat. Dalam
melaksanakan
penelitian
lebih
lanjut,
disarankan
juga
selain
menggunakan instrumen yang memuat
sejumlah penyataan dan pertanyaan terbuka
sebaiknya dilanjutkan dengan wawancara
yang lebih mendalam. 4. Hipotesis keempat yang menyatakan
bahwa motivasi intrinsik berpengaruh
positif dan signifikan terhadap kinerja
pegawai
terdukung
atau
diterima,
artinya
semakin
tinggi
motivasi
intrinsik yang dimiliki pegawai maka
kinerja
pegawai
akan
semakin
meningkat. Penggunaan model dalam penelitian
ini
menghasilkan
kontribusi
variable-
varaibel bebas yakni motivasi intrinsik,
komunikasi
interpersonal
dan
kepemimpinan transformasional sebesar
68.6 % terhadap kepuasan kerja, selain itu
variable-varaibel bebas yang terdiri dari
motivasi intrinsik, komunikasi interpersonal
dan kepemimpinan transformasional dan
kepuasan kerja sebesar terhadap kinerja
pegawai sebesar 78.2% terhadap kinerja
pegawai. Penambahan variabel - variabel
lain yang belum ada dalam penelitian yang
berkaitan dengan kepuasan kerja dan kinerja
pegawai dapat dilakukan untuk penelitian
lanjutan agar mendapatkan hasil yang lebih
baik. 5. Hipotesis kelima yang menyatakan
bahwa
komunikasi
interpersonal
berpengaruh positif dan signifikan
terhadap kinerja pegawai tidak terdung
atau ditolak, meskipun pengaruhnya
tidak signifikan tetapi menunjukan
pengaruh
yang
positif,
artinya
komunikasi interpersonal berpengaruh
positif namun tidak signifikan terhadap
kinerja pegawai. 6. Hipotesis keenam yang menyatakan
bahwa kepemimpinan transformasional
berpengaruh positif dan signifikan
terhadap terhadap kinerja pegawai 60 60 Volume 2 (1) - Juni 2018 Saran Impilikasi Manajerial Saran Impilikasi Manajerial
Berdasarkan
hasil
penelitian
diperoleh
bahwa
motivasi
intrinsik,
komunikasi
interpersonal
dan
kepemimpinan transformasional berdampak
positif terhadap kinerja pegawai. Sehingga
untuk meningkatkan kinerja pegawai dapat
dilakukan
dengan
cara
meningkatkan
motivasi intrinsik pegawai, komunikasi
interpersonal,
dan
kepemimpinan
transformasional secara langsung meapun
melalui kepuasan kerja. JRBM Tirtayasa meskipun
komunikasi
interpersonal
berpengaruh positif namun tidak memiliki
pengaruh yang berarti terhadap kinerja
pegawai. Berdasarkan sebagian tanggapan
responden, komunikasi interpersonal yang
terjalin belum berjalan efektif hal tersebut
dikarenakan anatar pegawai kurang adanya
keterbukaan
dalam
hal
penyamapai
informasi, terkadang hanya orang-orang
tertentu saja yang mendapatkan informasi
tersebut, sehingga menimbulkan prasangka-
prasangka yang tidak baik antar pegawai. Selain itu, terkadang ketika ada pegawai
yang mendapatkan masalah terutama dalam
hal pekerjaan, pegawai lainnya kurang
peduli dan kurang memiliki rasa empati
yang baik. Menyikapi permasalahan tersebut
agar komunikasi interpersonal berjalan
efektif, organisasi dalam hal ini Kantor
Kementerian Agama Kota Serang perlu
menciptakan suasana kerja yang akrab,
harmonis dan tidak terlalu kaku, contohnya
sesekali perlu diadakan kegiatan rihlah
(rekreasi) bersama sehingga diharapkan para
pegawai memiliki sikap positif, terbuka,
empati dan berbaik sangka terhadap sesama
rekan kerja, terhadap bawahan maupun
terhadap atasan. Salah satu hipotesis dalam penelitian
ini tidak terdukung atau ditolak, yaitu bahwa
komunikasi
interpersonal
berpengaruh
positif namun tidak signifikan terhadap
kinerja pegawai, sehingga memunculkan
gap dengan penelitian-penelitian terdahulu
yang
menyatakan
bahwa
komunikasi
interpersonal
berpengaruh
fositif
dan
signifikan
terhadap
kinerja
pegawai. Perbedaan
hasil
penelitian
ini
dapat
dijadikan research gap dalam penelitian
selanjutnya terutama yang berkaitan dengan
variabel komunikasi interpersonal terhadap
pemecahan masalah kinerja pegawai. penelitian
intrinsik,
dan Motivasi intrinsik yang dimiliki oleh
pegawai berdasarkan tanggapan responden
termasuk dalam kategori sedang, sehingga
perlu ditingkatkan lagi agar kinerja pegawai
semakin
meningkat. Hal
yang
dapat
dilakukan oleh organisasi dalam hal ini
Kantor Kementerian Agama Kota Serang
diantaranya adalah menyesuaikan target
sesui dengan kondisi dan kemampuan
pegawai
serta
kebutuhan
organisasi,
memberikan gambaran pola pengembangan
pegawai yang lebih jelas, menciptakan
suasana yang aman dan nyaman dalam
bekerja, dan organisasi perlu memberikan
penghargaan yang sesuai terhadap prestasi
kerja pegawai. Kepemimpinan
transformasional
yang memiliki pimpinan di Kementerian
Agama Kota Srang berdasarkan tanggapan
responden termasuk dalam kategori tinggi,
sehingga sosok dan sikap dari pemimpin ini
perlu dipertahankan, meskipun berada dalam
kategori
yang
tinggi
tetapi
masih
dimunkinkan untuk ditingkatkan lagi karena
sebagian kecil responden ada yang masih
beranggapan bahwa pimpinan di organisasi
tersebut
belum
sesuai
dengan
yang
diharapkan, misalnya terkesan pimpinan
membeda-bedakan
perlakuan
terhadap
bawahan,
kurang
rerspek
terhadap
permasalahan yang terjadi pada bawahan,
pimpinan kurang mampu dalam memotivasi
bawahan. Komunikasi
interpersonal
yang
terjalin diantara pegawai menurut tanggapan
responden termasuk dalam kategori sedang,
selain itu dilihat dari hasil uji signifikansi
menunjukan
bahwa
komunikasi
interpersonal bengaruh positif namun tidak
signifikan terhadap kinerja pegawai, artinya Kepuasan kerja yang dimiliki oleh
pegawai berdasarkan tanggapan responden
termasuk dalam kategori tinggi, sehingga
perlu dipertahankan. Meskipun berada
dalam kategori yang tinggi tetapi masih 61 Jurnal Riset Bisnis dan Manajemen Tirtayasa employee
performance
and
job
satisfaction in it park (software house)
sector
of
Peshawar,
Pakistan. International Journal of Academic
Research in Business and Social
Sciences. Vol. 6, No. 9. dimunkinkan untuk ditingkatkan lagi karena
sebagian kecil responden ada yang masih
beranggapan bahwa mereka belum merasa
puas dalam bekerja. Pimpinan organisasi
dapat mencipatakan suasan kerja yang aman,
nyaman, dan betah sehingga pegawai tidak
berpikir untuk pindah kerja atau pindah
ketempat lain, dan mereka merasa puas
bekerja ditempat merka bekerja saat ini. Ardana, K., Mujiati, N., dan Mudiarta, NW. 2012. Personal
management. Yogyakarta: Graha Ilmu Selanjutnya dari sisi kinerja pegawai
berdasarkan tanggapan responden termasuk
dalam kategori tinggi, sehingga perlu
diperthankan. Meskipun berada dalam
kategori
yang
tinggi
tetapi
masih
dimunkinkan untuk ditingkatkan lagi karena
sebagian kecil responden ada yang masih
beranggapan
bahwa
mereka
belum
menunjukan kinerja yang tinggi. Hal yang
dapat dilakukan oleh organisasi dalam hal
ini Kantor Kementerian Agama Kota Serang
diantaranya adalah menyesuaikan target
baik dari segi kuantitas, kulitas, ketepatan
waktu, efisiensi biaya, dan menetapkan
aturan sesuai dengan kemampuan dan
kondisi pegawai serta kebutuhan organisasi. penelitian
intrinsik,
dan Untuk meningkatkan kinerja pegawai selain
memperhatikan
indikasi
yang
mencerminkan
dari
variabel
kinerja
pegawai, yaitu dari segi kuantitas kerja,
kulitas kerja, ketepatan waktu dan efisiensi
biaya juga harus memperhatikan faktor-
faktor
yang
mempengaruhinya,
sebagaimana yang diajukan dalam penelitian
ini diantaranya; faktor motivasi intrinsik,
faktor
komunikasi
interpersonal,
kepemimpinan transformasional dan faktor
kepuasan kerja. Ariyabuddhiphongs, V dan Kahn, SI. 2017. Transformational
Leadership
and
Turnover Intention: The Mediating
Effects of Trust and Job Performance
on Café Employees in Thailand. Journal of Human Resources
in
Hospitality & Tourism. Vol. 16, No. 2, Arni, M. 2011. Komunikasi organisasi. Jakarta: Bumi Aksara Arthawan, KJ dan Mujiati, NW. 2017. Pengaruh
Gaya
Kepemimpinan
Transformasional dan Kepuasan Kerja
terhadap Kinerja Karyawan pada LPD
Kesiman
di
Denpasar. E-Jurnal
Manajemen Unud. Vol. 6, No. 3 Atambo, WN and Momanyi, DK. 2016. Effects of Internal Communication on
Employee Performance: A Case Study
of
Kenya
Power
and
Lighting
Company, South Nyanza Region,
Kenya. Imperial
Journal
of
Interdisciplinary
Research
(IJIR). Vol-2, Issue-5 Atambo, WN dan Ayaga, LK. 2016. The
Impact of Motivation on Employee
Performance: A Case Study of Health
Workers at Kisii Teaching and
Referral Hospital. Imperial Journal of
Interdisciplinary
Research
(IJIR). Vol-2, Issue-5 Daftar Pustaka Akhtar, W., Husnain, M., dan Akhtar, S. 2016. The
Impact
of
Intrinsik
Motivation
on
Employee’s
Job
Satisfaction. International Academy of
Science, Engineering and Technology:
Journal
of
Human
Resources
Management (IASET: JHRM). Vol. 1,
Issue 1, pp. 11-22 Babalola,
SS. 2016. The
Effect
of
Leadership Style, Job Satisfaction and
Employee-supervisor Relationship on
Job Performance and Organizational
Commitment. The Journal of Applied
Business Research, Vol. 32, Issue 3 Budianto dan Sambung, R. 2013. Pengaruh
Motivasi
Intrinsik
dan
Motivasi
Ekstrinsik terhadap Kinerja Pegawai
Melalui Kepuasan Kerja (Studi pada Ali, A., Bin, LZ., Piang, HJ dan Ali, Z. 2016. The impact of motivation on the 62 JRBM Tirtayasa Volume 2 (1) - Juni 2018 Volume 2 (1) - Juni 2018 Universitas Palangka Raya). JSM
(Jurnal Sains Manajemen): UNPAR. Vol.2 , No. 2 Effendy, OU. 2010. Komunikasi Teori dan
Praktek. Bandung:
Remaja
Rosdakarya. EK, K. dan Mukuru, E. 2013. Effect of
Motivation on Employee Performance
In Public Middle Level Technical
Training
Institutions
In
Kenya. International Journal of Advances in
Management and Economics. Vol.2,
Issue 4. Bungin,
B. 2010. Metode
Penelitian
Kuantitatif. Jakarta : Kencana Prenada
Media Group. Braun, S, Peus, C., Weisweiler, S dan Frey,
D. 2013. Transformational
Leadership, Job Satisfaction, and
Team Performance: A Multilevel
Mediation Model of Trust. The
Leadership Quarterly 24: Elsevier. Ferdinand, A. 2014. Metode Penelitian
Manajemen:
Pedoman
Penelitian
untuk Penulisan Skripsi, Tesis, dan
Disertasi Ilmu Manajemen. Semarang:
Badan
Penerbit
Universitas
Diponegoro Cheraghalizadeh, R. 2014. The Effect of
Intrinsik
Motivation
on
Job
Satisfaction:
Mediation
Role
of
Service
Recovery
Performance. International Journal of Economic
Practices and Theories. Vol. 4, No. 5,
Special Issue on Marketing and
Business Development. Fernadi, AD., Wahyu, D., dan Hidayati, N. 2016. Pengaruh Gaya Kepemimpinan
Transformasional, Motivasi Intrinsik
dan Motivasi Ekstrinsik terhadap
Kinerja Karyawan pada Perusahaan
Daerah
Air
Minum
(PDAM)
Kabupaten Tanah Laut. Jurnal Bisnis
dan Pembangunan. Vol 5, No. 2. Datt, R and Ram, S. 2014. Communication
Flow, Motivation Level and Job
Satisfaction
of
Self-Financing
Engineering
Colleges
by
Using
Mathematical Modeling. International
Journal of Research in Advent
Technology. Fini, FM dan Fini, AAS. 2015. A Study on
The
Relationship
Between
Communication Skills and Mental
Health
and
Job
Performance. Academic Journal of Psychological
Studies. Vol. 4, Issue 4. Dessler, G. 2011. Manajaemen Sumber
Daya Manusia. Jakarta: PT. Indeks Dhermawan, AANB., Sudibya, IGA dan
Utama,
WM. 2012. Pengaruh
Motivasi,
Lingkungan
Kerja,
Kompetensi,
dan
Kompensasi
terhadap Kepuasan Kerja dan Kinerja
Pegawai di Lingkungan Kantor Dinas
Pekerjaan Umum Provinsi Bali. Daftar Pustaka Jurnal
Manajemen, Strategi Bisnis, dan
Kewirausahaan. Vol. 6, 173 No. 2 Ghozali,
I, 2011, Structural Equation
Modeling Metode Alternatif Dengan
Partial Least Square (PLS) Edisi 3. Semarang: Badan Penerbit Universitas
Diponegoro Hair, J. F., Black, W. C., Babin, B. J., dan
Anderson, R. E. (2010). Multivariate
Data Analysis
(7th Edition). NJ:
Prentice Hall. Dirianzani, L., Sugiono dan Hardiningtyas,
D. 2014. Analisis Pengaruh Motivasi
Intrinsik dan Ekstrinsik terhadap
Kinerja Karyawan Borongan dengan
Komitmen
Organisasi
Sebagai
Mediasi (Study kasus pada perusahaan
rokok Adi Bungsu Malang). Jurnal
rekayasa dan manajemen sistem
industri Vol. 2 No. 5: Universitas
Brawijaya. Hakim, A., Yasir, M dan Nur, M. 2014. Pengaruh Hubungan Interpersonal dan
Lingkungan Kerja Terhadap Kepuasan
Kerja Perawat di Ruangan UGD RSID
Salewangang Maros. JurnalI lmiah
Kesehatan Diagnosisi. Vol. 4 No.5. Handoko, TH. 2010. Manajemen Personalia
dan
Sumber
Daya
Manusia. Yogyakarta: BPFE UGM. 63 Jurnal Riset Bisnis dan Manajemen Tirtayasa (Studi pada BTPN Syariah Semarang). Journal of Management. Vol.2 No.2 Hasibuan, MSP 2010. Manajemen Sumber
Daya Manusia, Edisi Revisi. Jakarta:
Bumi Aksara. Luthans, F. 2010. Perilaku Organisasi. Edisi 10. Yogyakarta: Andi. Hughes, RL., Ginnett, RC., Curphy, G.J. 2012. Leadership:
Memperkaya
Pelajaran dari Pengalaman. Edisi
Ketujuh. Jakarta: Salemba Humanika. Mangkunegara, AAP. 2011. Manajemen
Sumber Daya Manusia,
Bandung:
PT.Remaja Rosda Karya Ibrahim, MY. 2014. Model of E-leadership,
Intra-team Communication and Job
Satisfaction Among School Leaders in
Malaysia. Mediterranean Journal of
Social Sciences, Vol 5 No 23. ISSN
2039-2117. MCSER
Publishing,
Rome-Italy Marjianto. 2015. Pengaruh Komunikasi
Interpersonal
terhadap
Kinerja
Pegawai
Sekolah
Tinggi
Agama
Buddha Negeri (STABN) Raden
Wijaya Wonogiri Jawa Tengah. Jurnal
Widya Sandhi. Vol. 6 No. 1. Martoyo, S. 2010. Manajemen Sumber Daya
Manusia. BPFE. Yogyakarta. Jackson, JH. dan Robert LM. (2010). Human
Resource
Management. Ed.13th:
SouthWestern Cengage Learning. Mas’ud,
F. 2014. Survei
Diagnosis
Organisasional. Semarang:
Badan
Penerbit Universitas Diponegoro Jacobs, MA., Yu, W., Chavez, R. 2015. The
Effect of Internal Communication and
Employee Satisfaction on Supply
Chain
Integrasion. International
Journal of Production Economics. Masa'deh, R., Yousef, B dan Tarhini, OA. 2016. A Jordanian Empirical Study of
The
Associations
Among
Transformational
Leadership,
Transactional Leadership, Knowledge
Sharing, Job Performance, and Firm
Performance: A Structural Equation
Modelling Approach. Journal of
Management Development: Emerald
Insight. Vol. 35 Issue 5 Javed, M. 2014. Determinants of Job
Satisfaction
and
Its
Impact
on
Employee Performance and Turnover
Intentions. International Journal of
Learning & Development, Vol. 4, No. 2. Jogiyanto, HM. 2011. Konsep dan Aplikasi
Structural
Equation
Modeling
Berbasis Varian dalam Penelitian
Bisni. Daftar Pustaka Yogyakarta: UPP STIM YKPN Muhammad, R., Djudi, M dan Mayowan, Y. 2016. Pengaruh Gaya Kepemimpinan
Transformasional dan Transaksional
terhadap Kepuasan Kerja Karyawan
(Studi pada CV Kalingga Jaya di
Jakarta). Jurnal Administrasi Bisnis
(JAB), Vol. 35 No. 1 Linawati. 2014. Pengaruh Motivasi Intrinsik
dan
Motivasi
Kerja
Ekstrinsik
terhadap Kinerja Karyawan (Studi
pada PT. Angkasa Pura I Bandara
Udara Internasional Ahmad Yani
Semarang). Jurnal Kinerja. Vol. 18
No.1. Nasra, MA dan Heilbrunn, S. 2015. Transformational
Leadership
and
Organizational Citizenship Behavior
in the Arab Educational System in
Israel: The Impact Of Trust And Job
Satisfaction. Educational
Management
Administrasion
&
Leadership. Published by: SAGE Long, CS., Thean, LY., dan Kowang, TO. 2014. The
Transformational
Leadership: A Possible TQM Solution
to Increase Job Satisfaction?. Life
Science Journal. Vol. 10 No. 4. Nawawi, H. 2010. Manajemen Sumber Daya
Manusia
Untuk
Bisnis
Yang
Kompetitif. Yogyakarta : Gadjah Mada
Univesity Press. Lukito, HP., Haryono, AT dan Warso, MM. 2016. Pengaruh Motivasi Instrinsik,
Motivasi Ekstrinsik dan Pegalaman
Kerja Terhadap Kinerja Karyawan 64 JRBM Tirtayasa Volume 2 (1) - Juni 2018 Volume 2 (1) - Juni 2018 Northouse, PG. 2013. Teori dan Praktik
Kepemimpinan. Edisi
Keenam. Jakarta: Permata Putri. Satriowati, E., Paramita, PD, dan Hasiholan,
LB. (2016)
Pengaruh
Gaya
Kepemimpinan
Transformasional,
Kompensasi dan Komunikasi terhadap
Kinerja Karyawan dengan Kepuasan
Kerja sebagai Variabel Mediasi pada
Laundry Elephant King. Journal Of
Management, Vol. 2 No.2 Putra, IPOS dan Ardana, K. 2016. Pengaruh
Kepemimpinan Transformasional dan
Komitmen Organisasional terhadap
Kepuasan Kerja Karyawan pada PT. Kopi Bali Faktory Tour Denpasar. E-
Jurnal Manajemen Unud, Vol. 5, No. 5. Sedarmayanti. 2013. Manajemen Sumber
Daya Manusia. Bandung: Refika
Aditama. Putra, RB. 2016. Efek Mediasi Kepuasan
Kerja
terhadap
Kinerja
dengan
Komitmen Organisasi, Komunikasi
Interpersonal
dan
Organizational
Citizenship
Behaviors
sebagai
Variabel Anteseden. Jurnal Praktik
Bisnis. Vol.5 No. 1. Siagian, SP, 2011, Manajemen Sumber
Daya Manusia. Jakarta : Bumi Aksara. Cetakan kesembilan belas Siengthai, S dan Pila-Ngarm, P. 2016. The
Interaction Effect of Job Redesign and
Job
Satisfaction
on
Employee
Performance, Evidence-based HRM: a
Global
Forum
for
Empirical
Scholarship, Vol. 4 Issue 2. Published
by Emerald Insight. Raina, R and Roebuck, DB. 2014. Exploring
Cultural Influence on Managerial
Communication in Relationship to Job
Satisfaction,
Organizational
Commitment, and the Employees'
Propensity to Leave in the Insurance
Sector of India. International Journal
of
Business
Communication. Published by: SAGE Sugiyono. 2013. Metode
Penelitian
Manajemen. Bandung : Alfabeta Tania,
Y. 2017. Pengaruh
Gaya
Kepemimpinan Transformasional dan
Motivasi Kerja terhadap Kinerja
Karyawan
pada
PT. Daftar Pustaka Premier
Management
Consulting. Jurnal
Arsitektur
AGORA:
Universitas
Trisakti. Vol. 5, No. 1 Raza, MY., Roebuck, MWA., Husnain, M
dan Akhtar, MS. 2015. The Impact of
Intrinsik Motivation on Employee’s
Job Satisfaction. Journal Management
and Organizational Studies. Vol. 2,
No.3 Tan, TH dan Waheed, A. (2011). Herzberg’s
motivation-hygiene theory and job
satisfaction in the Malaysian retail
sector: Mediating effect of love of
money. Asian
Academy
of
Management Journal, No.16 Rivai,
V
dan
Basri,
AFM. 2010. Performance appraisal. Sistem yang
tepat untuk menilai kinerja karyawan
dan
Meningkatkan
daya
saing
perusahaan. Jakarta:
PT. Raja
Grafindo Persada Thamrin, HM. 2012. The Influence of
Transformational
Leadership
and
Organizational Commitment on Job
Satisfaction
and
Employee
Performance. International Journal of
Innovation,
Management
and
Technology, Vol. 3 Rivai, V., Ramly, M., Mutis, T dan Willy A. 2014. Manajemen Sumber Daya
Manusia untuk Perusahaan dari Teori
ke Praktek. Jakarta : PT. Raja Grafindo
Persada Robbins, SP. 2010. Prinsip-prinsip Perilaku
Organisasi. Edisi Kelima. Jakarta:
Erlangga Tika, MP. (2010). Budaya Organisasi dan
Peningkatan Kinerja Perusahaan. PT
Bumi Aksara. Jakarta. Robbins, SP dan Judge, TA. 2015. Perilaku
Organisasi. Edisi 16. Jakarta: Salemba
Empat. Taufik,
A. 2016. Pengaruh
Motivasi
Intrinsik terhadap Kinerja Karyawan 65 Jurnal Riset Bisnis dan Manajemen Tirtayasa dengan Kepuasan Kerja dan Disiplin
Kerja sebagai Variabel Mediasi pada
Badan Pusat Statistik Kabupaten
Magelang. Jurnal Bisnis: Teori dan
implementasi. Vol. 7 No. 2 Usman, B. 2013. Pengaruh Komunikasi
Interpersonal
Terhadap
Kinerja
Pegawai pada Fakultas Ekonomi
Universitas PGRI Palembang. Jurnal
Media Wahana Ekonomika. Vol. 10
No.1. Utami, SP. 2015. Pengaruh Kepemimpinan,
Motivasi
dan
Komunikasi
Interpersonal
terhadap
Kinerja
Karyawan
pada
Biro
Perjalanan
Wisata Panen Tour. Jurnal Ilmiah
WIDYA. Vol. 3 No. 1, ISSN: 2337-
6686 Wexley, KN. dan Yukl, GA. 2010. Perilaku
Organisasi dan Psikologi Personalia. Terjemahan
Shobaruddin
Muh. Jakarta: PT. Asdi Mahasatnya. Yuki, AG. 2010. Kepemimpinan dalam
Organisasi. Edisi 5 Alih Bahasa Budi
Suprianto. Jakarta: Indeks 66 66 Halaman ini sengaja dikosonngkan 67 67
|
https://openalex.org/W2384171469
|
https://orca.cardiff.ac.uk/id/eprint/89173/8/jad%252F2016%252F53-1%252Fjad-53-1-jad160146%252Fjad-53-jad160146.pdf
|
English
| null |
Immunization Against Specific Fragments of Neurotrophin p75 Receptor Protects Forebrain Cholinergic Neurons in the Olfactory Bulbectomized Mice
|
Journal of Alzheimer's disease
| 2,016
|
cc-by
| 8,776
|
289 289 Journal of Alzheimer’s Disease 53 (2016) 289–301
DOI 10.3233/JAD-160146
IOS Press 1These authors contributed equally to this work.
∗Correspondence to: Dr. V. Vorobyov, PhD, Institute of Cell
Biophysics, Pushchino, 142290, Russia. Tel.: +7 4967 739 223;
Fax: +7 4967 330 509; E-mail: vorobyovv2@gmail.com. Accepted 31 March 2016 Abstract. Alzheimer’s disease (AD) is characterized by progressive cognitive impairment associated with marked cholinergic
neuron loss and amyloid- (A) peptide accumulation in the brain. The cytotoxicity in AD is mediated, at least in part, by A
binding with the extracellular domain of the p75 neurotrophin receptor (p75NTR), localized predominantly in the membranes
of acetylcholine-producing neurons in the basal forebrain. Hypothesizing that an open unstructured loop of p75NTR might
be the effective site for A binding, we have immunized both olfactory bulbectomized (OBX) and sham-operated (SO)
mice (n = 82 and 49, respectively) with synthetic peptides, structurally similar to different parts of the loops, aiming to
block them by specific antibodies. OBX-mice have been shown in previous studies, and confirmed in the present one, to be
characterized by typical behavioral, morphological, and biochemical AD hallmarks, including cholinergic deficits in forebrain
neurons. Immunization of OBX- or SO-mice with KLH conjugated fragments of p75NTR induced high titers of specific
serum antibodies for each of nine chosen fragments. However, maximal protective effects on spatial memory, evaluated in a
Morris water maze, and on activity of choline acetyltransferase in forebrain neurons, detected by immunoreactivity to specific
antibodies, were revealed only for peptides with amino acid residue sequences of 155–164 and 167–176. We conclude that the
approach based on immunological blockade of specific p75NTR sites, linked with the cytotoxicity, is a useful and effective
tool for study of AD-associated mechanisms and for development of highly selective therapy of cholinergic malfunctioning
in AD patients. Keywords: Acetyltransferase, amyloid-beta, cholinergic mediation, Morris water maze, olfactory bulbectomy, spatial memory Keywords: Acetyltransferase, amyloid-beta, cholinergic mediation, Morris water maze, olfactory bulbectomy, spatial memory Keywords: Acetyltransferase, amyloid-beta, cholinergic mediation, Morris water maze, olfactory Immunization Against Specific Fragments
of Neurotrophin p75 Receptor Protects
Forebrain Cholinergic Neurons
in the Olfactory Bulbectomized Mice Natalia Bobkovaa,1, Vasily Vorobyova,1,∗, Natalia Medvinskayaa, Inna Nesterovaa,
Olga Tatarnikovaa, Pavel Nekrasova, Alexander Samokhina, Alexander Deevb, Frank Sengpielc,
Dmitry Koroevd and Olga Volpinad Natalia Bobkovaa,1, Vasily Vorobyova,1,∗, Natalia Medvinskayaa, Inna Nesterovaa,
Olga Tatarnikovaa, Pavel Nekrasova, Alexander Samokhina, Alexander Deevb, Frank Sengpielc,
Dmitry Koroevd and Olga Volpinad aInstitute of Cell Biophysics, Russian Academy of Sciences, Pushchino, Moscow Region, Russia
bInstituteofTheoreticalandExperimentalBiophysics,RussianAcademyofSciences,Pushchino,MoscowRegion,
Russia cSchool of Biosciences and Neuroscience & Mental Health Research Institute, Cardiff University, Museum
Avenue, Cardiff, UK
d dShemyakin’s-Ovchinnikov’s Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia Handling Associate Editor: Vladimir Buchman 1These authors contributed equally to this work. ISSN 1387-2877/16/$35.00 © 2016 – IOS Press and the authors. All rights reserved words: Acetyltransferase, amyloid-beta, cholinergic mediation, Morris water maze, olfactory bulbectomy, s INTRODUCTION These findings suggest that a potential
treatment for AD should be targeted at an early stage
of the disease progression [4]. The most suitable
model directly associated with AD progression is
that of animals with surgically removed olfactory
bulbs (OBX) that showed both typical behavioral,
morphological, and biochemical AD hallmarks [5]
and phasic time courses in their expression after the
bulbectomy [6]. One of potential targets thought to
be critically involved in the pathogenesis of AD, a
neurotrophin cell surface receptor (NTR), p75NTR
[7], has been shown to bind with amyloid- (A)
peptides [8] that, in turn, promotes A-associated
neuronal dystrophy [9], and regulates both A levels
[10, 11], neuronal death [12], and cholinergic trans-
mission [13]. Furthermore, in the AD brain, p75NTR
expression is enhanced both in forebrain neurons
and in their terminals in the cortex and hippocampus
[14, 15]. Interestingly, the A-induced neurotoxicity
has been noted to be attenuated in these brain areas
whereas the loss of cholinergic neurons in the basal
forebrain to be increased [16]. The multifunctional
characteristics of p75NTRs seem to be associated
with the capability of A-p75NTR complexes to
activate a number of extra- and intracellular sig-
naling pathways [17], implementing selectivity of
A-produced neurotoxicity toward cholinergic basal
forebrain neurons in particular [18, 19]. Within the
intracellular structures activated through extracel-
lular domain of p75NTR, a death receptor (DR6)
is associated with A-induced apoptosis [7], and
its blockade by specific antibodies (“intrabodies”)
has been shown to attenuate A-induced caspase
3 activation mediated by DR6 and resulting in cell
death through the Jun kinase (JNK) pathway [20]. The neurotrophin binding with the extracellular
domain of p75NTR may promote neuronal survival
as well, either through the nuclear factor-B (NF-B)
pathway or via p75NTR crosstalk with an “anti-
i ”
i
l
d ki
(T k) In our present study with nine selected amino acid
fragments of p75NTR, only immunization against
two of sequences 155–164 and 167–176 effectively
protected cholinergic mediation in the forebrain and,
as a result, cognitive abilities of OBX-mice from
AD-like neurodegeneration. INTRODUCTION Within the
intracellular structures activated through extracel-
lular domain of p75NTR, a death receptor (DR6)
is associated with A-induced apoptosis [7], and
its blockade by specific antibodies (“intrabodies”)
has been shown to attenuate A-induced caspase
3 activation mediated by DR6 and resulting in cell
death through the Jun kinase (JNK) pathway [20]. The neurotrophin binding with the extracellular
domain of p75NTR may promote neuronal survival
as well, either through the nuclear factor-B (NF-B)
pathway or via p75NTR crosstalk with an “anti-
apoptotic” tropomyosin related kinase (Trk) receptor
[21]. The expression of Trk subtype NTR has been
shown to be decreased in basal forebrain cholinergic
neurons in AD [22]. Nevertheless, conformational
transformations revealed after neurotrophin binding with p75NTR and favorable to simultaneous binding
of Trk and p75NTR monomers with the same neu-
rotrophin dimer [23], might be a reasonable basis for
development of a potentially effective approach to
treat the AD-associated cholinergic malfunctioning. Indeed, the cysteine-rich (CR1-4) repeats of the
p75NTR extracellular domain have been shown to
have several binding sites for neurotrophins and A
[7, 24]. Therefore, we suggest that the open unstruc-
turedloopsinthisdomaincontainthesitesforbinding
with ligands and are the most accessible targets for
immunological treatment. Recently, an antibody
against the extracellular domain of p75NTR has been
shown to inhibit, partially, A1–42-induced cell death
in hippocampal neuronal cultures in vitro [18]. To
unmask in vivo the p75NTR site(s) specifically linked
with loss of ChAT-containing forebrain neurons typi-
cal for AD, we immunized OBX-mice with synthetic
peptides, analogs of different p75NTR fragments
of these loops. In addition, they were characterized
by relatively high homology of their primary amino
acid structures in mouse and human. To evaluate the
efficacy of the treatment in OBX-mice, the extents of
both spatial memory improvement in a Morris water
maze and recovery of ChAT-positive cell density
in the basal forebrain were analyzed. Recently, this
immunological approach has allowed the revealing
of the A-binding sites on 7-nicotinic acetylcholine
and prion receptors whose blockade with specific
antibodies suppressed the progression of AD-linked
neurodegeneration in OBX mice [25, 26]. associated
with
impairment
of
cognition
in
Alzheimer’s disease (AD) in particular [1]. AD
progression has been shown to be characterized by
the attenuating activity of choline acetyltransferase
(ChAT), an acetylcholine-synthesizing enzyme [2]. However, at preclinical stages of AD, ChAT activity
was unaffected, which is supposed to be associated
with
involvement
of
cholinergic
compensatory
plasticity [3]. INTRODUCTION 1These authors contributed equally to this work. ∗Correspondence to: Dr. V. Vorobyov, PhD, Institute of Cell
Biophysics, Pushchino, 142290, Russia. Tel.: +7 4967 739 223;
Fax: +7 4967 330 509; E-mail: vorobyovv2@gmail.com. The basal forebrain cholinergic neurons, innervat-
ingthecerebralcortexandhippocampus,areinvolved
in learning/memory functions, and their loss is ISSN 1387-2877/16/$35.00 © 2016 – IOS Press and the authors. All rights reserved ed online with Open Access and distributed under the terms of the Creative Commons Attribution License (CC-BY). This article is published online with Open Access and distributed under the terms of the Creative Commons Attribution N. Bobkova et al. / p-75NTR specificity and cholinergic mediation in AD 290 associated
with
impairment
of
cognition
in
Alzheimer’s disease (AD) in particular [1]. AD
progression has been shown to be characterized by
the attenuating activity of choline acetyltransferase
(ChAT), an acetylcholine-synthesizing enzyme [2]. However, at preclinical stages of AD, ChAT activity
was unaffected, which is supposed to be associated
with
involvement
of
cholinergic
compensatory
plasticity [3]. These findings suggest that a potential
treatment for AD should be targeted at an early stage
of the disease progression [4]. The most suitable
model directly associated with AD progression is
that of animals with surgically removed olfactory
bulbs (OBX) that showed both typical behavioral,
morphological, and biochemical AD hallmarks [5]
and phasic time courses in their expression after the
bulbectomy [6]. One of potential targets thought to
be critically involved in the pathogenesis of AD, a
neurotrophin cell surface receptor (NTR), p75NTR
[7], has been shown to bind with amyloid- (A)
peptides [8] that, in turn, promotes A-associated
neuronal dystrophy [9], and regulates both A levels
[10, 11], neuronal death [12], and cholinergic trans-
mission [13]. Furthermore, in the AD brain, p75NTR
expression is enhanced both in forebrain neurons
and in their terminals in the cortex and hippocampus
[14, 15]. Interestingly, the A-induced neurotoxicity
has been noted to be attenuated in these brain areas
whereas the loss of cholinergic neurons in the basal
forebrain to be increased [16]. The multifunctional
characteristics of p75NTRs seem to be associated
with the capability of A-p75NTR complexes to
activate a number of extra- and intracellular sig-
naling pathways [17], implementing selectivity of
A-produced neurotoxicity toward cholinergic basal
forebrain neurons in particular [18, 19]. Spatial discrimination testing (Morris water
maze) The first immunization procedure (Day 1) initi-
ating production of antibodies against one of nine
selected p75NTR fragments (32–46, 39–47, 66–72,
86–93, 97–105, 115–122, 147–154, 155–164, and
167–176) was performed on 2-month old mice, which
were then bulbectomized or sham-operated on Day
25 and immunized repeatedly on Day 43. For the
immunization, emulsions of PBS solution of a KLH-
peptide conjugate (Sigma-Aldrich, USA) mixed with
either CFA or IFA (the 1st or the 2nd immunization,
respectively; Disco laboratories, USA) were subcu-
taneously (s.c.) injected at a dose of 100 g of the
protein per mouse in the proximal part of its tail (in
control groups, vehicle was used). Starting on Day
53 mice were trained in a Morris water maze: Ini-
tially, in 3 trials, to reach a visible saving platform
and then, in 4 daily trials for 5 days, with an invisible
(submerged) platform. Finally, on Day 58, a mem-
ory test followed by the brain preparation for further
histological and histoimmunochemical analysis was
performed. The serum samples for the antibody titer
estimation were taken from the eye vein on Day 52
and kept at –20◦C for further analysis (see results
in Table 1). As controls for the p75NTR fragments,
KLH (Keyhole Limpet Hemocyanin) conjugate of
SQDRHKQKIIAPAKQLL peptide corresponding to
the fragment of 197–213 of VP1 protein from foot-
and-mouth disease virus (VP1), and KLH along
were used. The Maxisorp’s 96-wells plates (Nunc,
Denmark) were coated with 20 g/ml ovalbumin-
conjugated peptide in 0.05 M sodium bicarbonate
buffer (pH 9.5); blood serum samples in successive On the 28th day after surgery (Day 53 after 1-st
immunization), the learning/memory abilities of the
mice were tested with the Morris water maze in a cir-
cular water pool (90 cm in diameter, 50 cm in height),
filled to a depth of 40 cm with water (33◦C±1) ren-
dered opaque by the addition of powdered milk. Four
equal virtual sectors in the pool were arbitrary indi-
catedasI,II,III,andIV.Eachmousewasfirsttestedon
its capability to reach a visible platform (0.5 cm above
the water surface, 8 cm in diameter). All mice used
in this study demonstrated similar swimming abili-
ties in reaching the visible platform. A day later and
for 5 days, each mouse was trained (4 trials per day,
with an intertrial interval of 25–30 min) to find the
platform centered in sector III but submerged 0.5 cm
below the surface. Olfactory bulbectomy (OBX) twofold dilutions were added. Then the incubation
with anti-mouse immunoglobulin antibodies conju-
gated to horseradish peroxidase was carried out. The
substrate solution (0.05% hydrogen peroxide and
0.05% o-phenylenediamine in citrate buffer, pH 5.0)
was added to the wells for visualization. The reaction
was stopped by adding 10% sulfuric acid, and opti-
cal absorption was measured at 492 nm. The highest
dilution of the sera with OD >0.1, which was still
positive and exceeded the level of preimmune serum
more than twofold, was taken as the antibody titer. Nearly 3-month-old mice (25–30 g) were anaes-
thetized with Nembutal (40 mg/kg, i.p.) and a hole of
2 mm in diameter was drilled medially over the olfac-
tory bulbs (2.0 mm anterior to bregma). The bulbs
were aspirated carefully under visual control through
a blunt needle attached to a water pump. Sham-
operated mice were treated similarly with exception
of the bulbectomy. MATERIAL AND METHODS One hundred and thirty one male mice (NMRI,
Charles River, USA) bred in a colony under con-
trolled barrier conditions were used in this study. The animals were given food and water ad libitum
and reared in a standard 12-h/12-h day/night cycle. The procedures were carried out in accord with the
principles enunciated in the Guide for Care and Use
of Laboratory Animals, NIH publication No 85-23,
and approved by the Institutional Ethical Review
Committee. N. Bobkova et al. / p-75NTR specificity and cholinergic mediation in AD 291 Histology/morphology and choline
acetyltransferase (ChAT) evaluation tetra-hydrochloride and 0.02% H2O2 in PBS at 20◦C. Finally, these slices were putted into the Apati’s
media followed by their examination at 40-x magnifi-
cation on an optical microscope Nikon Eclipse E200
with digital DXM1200 camera. The images of slices
with NMB were stored for further quantification
of ChAT-immunopositive neurons. To evaluate the
extent of the bulbectomy-produced damage in NBM
a neuronal apoptosis was analyzed by use of two
sorts of staining: Conventional Nissl, for the whole
cells, and DAPI, demonstrating nuclear chromatin
destruction that is directly associated with apoptosis. In DADL analysis, only roundish intracellular nuclei
with size of >10 m, typical for neurons, were used. The number of apoptotic neurons versus total number
of cells in 3-4 brain slices with NBM was estimated
and presented in percentage for each of the p75NTR
fragments used. Immediately after memory test session, all mice
were deeply anaesthetized with overdose of Nem-
butal (60 mg/kg, i.p., Sigma, USA), their brains
were perfused through the left cardiac ventricle with
a phosphate-buffered solution and verified on the
extent of the bulbectomy. Only the brains with both
bulbs fully removed (>90% of their estimated vol-
umes) and no damage to prefrontal cortices were
accepted for further histological determination of
a population ratio of pathological versus normal
neurons and immunohistochemical analysis of the
basal forebrain ChAT-containing cells. The animals
with insufficient bulbectomy were excluded from
further analysis of results of their learning/memory
abilities. The brain blocks containing the cortex and hip-
pocampus, preliminarily fixed in 4% paraformalde-
hyde (dissolved in 0.1 M PBS, pH 7.4) for 24 h
at RT, were used for morphological and immuno-
histochemical evaluation. Each block was cut on
a freezing microtome (Reicher, Austria) into 10-
m and 30-m coronal sections collected at 70-m
intervals. The thin (10-m) slices were stained with
0.1% Cresyl Violet, and their magnified (at 20x
or 40x, optical microscope Nikon Eclipse E200)
and digitized (DXM1200 camera) images were used
for further morphological and morphometric exam-
inations. All neurons were divided into a normal
category and several pathologies characterized by
cytolysis, karyolysis, pyknosis, and vacuolization. The neurons from different categories were counted
in each investigated brain areas as a percentage of
total cell population observed in 10–12 randomly
selected fields (100 cells in each fields) in 3-4 sections
per mouse. Spatial discrimination testing (Morris water
maze) The animals were placed in a ran-
domly chosen sector and allowed a maximum of one
minute to find the platform followed by a 15-s rest
period, once on the platform, before being dried and
relocated into the home cage. An unsuccessful mouse
was gently redirected to the platform after exceeding
the 60-s limit. On the day after the end of the training
trials,a60-smemorytest,withtheplatformtemporar-
ily removed from the pool, was given to assess spatial
memory in mice for the platform’s former location
(in the sector III). The total time spent in the target
sectorIIIduringthetestwerecomputedusingahome-
developedtrackingsystembyusingofadigitalcamera
(Logitech QuickCam 3000, 800 × 600 pixels, 15 fps)
located 90 cm above the pool. Table 1
Serum antibody titers in the olfactory-bulbectomized mice immunized against different synthetic p75 fragments conjugated
with the KLH-peptide
Serum antibody titers (in -lg of sera dilution) versus p75NTR fragments
P75NTR fragment
32–46
39–47
66–72
86–93
97–105
115–122
147–154
155–164
167–176
Antibody titer
3.8–4.1
3.4–4.1
3.1–3.4
3.1–3.4
3.7–4.1
2.2–2.8
2.2–2.5
3.1–3.8
3.4–4.1 N. Bobkova et al. / p-75NTR specificity and cholinergic mediation in AD 292 Peptide synthesis Peptides were synthesized using reagents and
aminoacidderivativesproducedbyMerck(Germany)
andFluka(Switzerland).Immunologicalstudieswere
carried out with the keyhole limpet hemocyanin
(KLH) solution in PBS at a concentration of 5 mg/ml
(Sigma–Aldrich, USA), 25% aqueous solution of
glutaric dialdehyde (Sigma, USA), Freund’s com-
plete adjuvant, Freund’s incomplete adjuvant (Difco
Laboratories, USA), goat antimouse immunoglobu-
lin antibodies conjugated to horseradish peroxidase
(Imtek, Russia), and 96-well Maxisorp plates (Nunc,
Denmark). Peptides were chosen according to the
sequence of the human p75 (TNR16 HUMAN)
UniProtKB), and their synthesis was carried out on
Wang resin, using the Fmoc/But-scheme. All pep-
tidescontainedadditionalC-terminalglycineresidue. TBTU/ DIEA method was applied for elongation of
peptidechain.Cleavageofpeptidesfromtheresinwas
carried out in a mixture of TFA (94%), triisopropy-
lsilane (1%), 1,2-ethane dithiol (2.5%), and water
(2.5%) for 2 h. The peptides were purified by reverse
phase HPLC in Phenomenex Jupiter 10 C18 300A
250×10 mm in an acetonitrile gradient from 10 to
70% in 0.1% TFA solution. Synthetic peptides were
characterized by the data of amino acid analysis, ana-
lytical HPLC, and mass spectrometry. The thick (30-m) slices were used for the analysis
of the ChAT-containing cells in the nucleus basalis
magnocellularis (NBM) [27], the equivalent of the
primate n. basalis of Meynert [28]. The slices were
incubated in PBS with consecutive addition of a) 1%
H2O2, for 30 min; b) 5% bowing serum albumin,
for 2 h; c) polyclonal rabbit antibodies against ChAT
(ab68799, Abcam, Cambridge, USA) with 1:200
dilution, for 12 h at 4◦C; d) biotinylated coat antibod-
ies against rabbit’s -globulines with 1:100 dilution,
for2 hat20◦C;e)avidine-biotinecomplexwith1:100
dilution, for 1 h at 20◦C. The avidine-biotine com-
plex’s peroxidase activity was revealed by incubation
of slices with both 0.5% 3,3’- diamino-benzidin- Peptide conjugates with KLH and ovalbumin The KLH-conjugated peptides were prepared as
follows: 1 mg of the peptides was added to 1 ml N. Bobkova et al. / p-75NTR specificity and cholinergic mediation in AD 293 After the immunization with synthetic analogs of the
p75NTR fragments, only four of them with residuals
of32–46,86–93,155–164,and167–176werestatisti-
cally effective in protecting the cholinergic cells from
their depletion observed in non-immunized OBX-
mice, with maximal effects associated with peptides
of 155–164 and 167–176 (Fig. 1A). This is clearly
visible on individual slices with enhanced densities of
ChAT-containing neurons in NBM in two OBX-mice,
immunized against 155–164 and 167–176 fragments
of p75NTR (c.f., Fig. 1E and F versus D, respectively)
and confirmed by one-way ANOVA (F10,487 = 74.6,
p < 0.001) for the whole set of slices. of KLH solution in PBS (5 mg/ml, pH 7.4) and
stirred for 30 min. Then 50 l of 0.25% glutaralde-
hyde solution was added by drops, incubated for 16 h,
and dialyzed against PBS for 18 h, with the buffer
changed three times. The ovalbumin conjugate was
prepared by dissolving 1 mg of the peptide in 500 l
of PBS, with the addition of 2 mg of ovalbumin. Then
50 l of hydrochloride N-(3-dimethylaminopropyl)-
N’-ethylcarbodiimide solution (0.08 mg/ml) was
added by drops during 30 min, incubated under stir-
ring (16 h) and dialyzed against PBS with the buffer
changed two times. Statistics In yhe NBM area, increased amount of apop-
totic cells was found (Fig. 2) in brain slices from
OBX- versus SO-mice (n = 136/25 versus 18/7,
respectively) that was significant regardless whether
the data from all OBX-mice or only from treated
with
p75NTR
fragments
were
used
(one-way
ANOVA: F10,143 = 12.2 or F9,132 = 10.8, respectively,
p < 0.001, for both). However, the immunized versus
non-immunized OBX-mice demonstrated significant
shrinking of apoptotic cell populations (one-way
ANOVA: F9,126 = 7.9, p < 0.001) with evident bene-
ficial effect for p75NTR fragments of 39–47, 66–72,
97–105, 155–164, and 167–176 (16/3, 16/3, 16/3,
12/3, and 12/3 slices/mice, respectively). In contrast
to these, the treatments with 32–46, 86–93, 115–122,
and 147–154 (16/4, 12/3, 8/2, and 16/3 slices/mice,
respectively) were practically ineffective. Whether
such immunization is beneficial for other brain struc-
tures, affected by bulbectomy, is analyzed in the next
section. Differences in behavioral, immunohistochemi-
cal, and morphometric parameters were evaluated
by either non-parametric Mann-Whitney U-test,
one/two-way ANOVA for repeated measures or two-
tailed Student’s t-test, when appropriate (p < 0.05 was
deemed statistically significant). The data in the text
and figures are expressed as mean ± SEM. Selective effects of immunization against different
p75NTR fragments on acetylcholine-synthesized
capability of neurons in the nucleus basalis
magnocellularis (n. Meynert) Selective effects of immunization against different
p75NTR fragments on acetylcholine-synthesized
capability of neurons in the nucleus basalis
magnocellularis (n. Meynert) To test the idea of possible brain protection against
amyloid-produced toxicity by use of an immunolog-
ical blockade of A binding with the neurotrophin
receptors, we have studied forebrain cholinergic
neuron functioning in the olfactory-bulbectomized
(OBX) mice, which have been shown to be char-
acterized by enhanced A levels in the brain [29]. In this section, we analyzed 498 brain slices from
48 mice distributed within the groups as follows
(slices/mice): SO (61/7), OBX (86/6) or OBX after
theimmunizationwithsyntheticpeptides,theanalogs
of the p75NTR fragments. They were the fol-
lowing (slices/mice): 32–46 (94/6), 39–47 (25/3),
66–72 (24/3), 86–93 (46/4), 97–105 (44/4), 115–122
(27/3),147–154(35/4),155–164(24/4),and167–176
(32/4). In the immunohistochemical staining of the
brain slices by use of specific antibodies against
choline acetyltransferase (ChAT), a key enzyme for
acetylcholine synthesis, significant depletion of the
neuronal population of the forebrain ChAT-positive
neurons in NBM was revealed in OBX- versus SO-
mice one month after the bulbectomy (Fig. 1A). Morphological and functional characteristics
of cortical and hippocampal neurons after
immunization against different p75NTR
fragments Morphological and functional characteristics
of cortical and hippocampal neurons after
immunization against different p75NTR
fragments Recently, in OBX-mice, demonstrating typical AD
hallmarks, pathological changes in the cortical and
hippocampal neurons, closely associated with learn-
ing and memory processing, have been revealed
[26]. The development of these cellular changes
was characterized by nuclear fading (karyolysis)
or shrinkage (pyknosis), membrane disintegration
(cytolysis), intracellular vacuole formation (vac-
uolization), and by decreased neuronal density,
resulting in neuronal death (Fig. 3A). Statistical anal-
ysis of these parameters in OBX-mice has revealed a
predominantly protective influence of the immuniza-
tion against the p75NTR fragments on the survival (32/4). In the immunohistochemical staining of the
brain slices by use of specific antibodies against
choline acetyltransferase (ChAT), a key enzyme for
acetylcholine synthesis, significant depletion of the
neuronal population of the forebrain ChAT-positive
neurons in NBM was revealed in OBX- versus SO-
mice one month after the bulbectomy (Fig. 1A). 294 N. Bobkova et al. / p-75NTR specificity and cholinergic mediation in AD Detrimental effect of olfactory bulbectomy (OBX) in mice on the density of choline acetyltransferase (ChAT)-cont
orebrain, beneficial effects of the immunization against the p75NTR amino acid residues of 32–46, 86–93, and
wing effects of the treatment with 155–164 and 167–176 fragments. The immunoreactive labeling against ChAT
magnocellularis on the representative examples (taken from the area marked by a rectangle on B) demonstrates dec Fig. 1. Detrimental effect of olfactory bulbectomy (OBX) in mice on the density of choline acetyltransferase (ChAT)-containing neurons
in the forebrain, beneficial effects of the immunization against the p75NTR amino acid residues of 32–46, 86–93, and 147–154, and
overgrowing effects of the treatment with 155–164 and 167–176 fragments. The immunoreactive labeling against ChAT in the nucleus
basalis magnocellularis on the representative examples (taken from the area marked by a rectangle on B) demonstrates decreased number
of the labeled cells after the bulbectomy (c.f., C and D) and their recovery after the immunization against the fragments of 155–164 (E)
and 167–176 (F). White horizontal bars in C-F are the scale bars of 50 m. On A, error bars show 1 SEM; star or plus denote significant
differences versus either SO or OBX, respectively (one and two markers are p < 0.05 and 0.01, Mann-Whitney U-test). The brain slice on B
was modified from [40]. Fig. 1. Morphological and functional characteristics
of cortical and hippocampal neurons after
immunization against different p75NTR
fragments Detrimental effect of olfactory bulbectomy (OBX) in mice on the density of choline acetyltransferase (ChAT)-containing neurons
in the forebrain, beneficial effects of the immunization against the p75NTR amino acid residues of 32–46, 86–93, and 147–154, and
overgrowing effects of the treatment with 155–164 and 167–176 fragments. The immunoreactive labeling against ChAT in the nucleus
basalis magnocellularis on the representative examples (taken from the area marked by a rectangle on B) demonstrates decreased number
of the labeled cells after the bulbectomy (c.f., C and D) and their recovery after the immunization against the fragments of 155–164 (E)
and 167–176 (F). White horizontal bars in C-F are the scale bars of 50 m. On A, error bars show 1 SEM; star or plus denote significant
differences versus either SO or OBX, respectively (one and two markers are p < 0.05 and 0.01, Mann-Whitney U-test). The brain slice on B
was modified from [40]. N. Bobkova et al. / p-75NTR specificity and cholinergic mediation in AD 295 Fig. 2. Increase of the apoptotic cell population in the nucleus basalis magnocellularis in OBX- versus SO-mice (OBX+V versus SO+V) and
neuronal protective effect of the immunization against p75NTR fragments of 39–47, 66–72, 97–105, 155–164, and 167–176. Stars denote
significant (p < 0.01, Mann-Whitney U-test) differences versus OBX. Error bars show 1 SEM. Fig. 2. Increase of the apoptotic cell population in the nucleus basalis magnocellularis in OBX- versus SO-mice (OBX+V versus SO+V) and
neuronal protective effect of the immunization against p75NTR fragments of 39–47, 66–72, 97–105, 155–164, and 167–176. Stars denote
significant (p < 0.01, Mann-Whitney U-test) differences versus OBX. Error bars show 1 SEM. observed after the immunization against 86–93 and
147–154 p75NTR fragments (Fig. 3C, sectors 4
and 7, respectively) and were expressed in both
decreased population of healthy neurons (Fig. 3C, a)
and increased ones of pathological cells with visi-
ble cytolysis, vacuolization, and karyolysis (Fig. 3C,
b-d). The changes in CA3 are especially impressive
as they were observed at relatively low reference
level of neuronal density in this area (Fig. 3A, f). Theimmunologicaltreatmentwith155–164p75NTR
fragment resulted in shrinkage of the karyolitic
cell population both in the cortex and hippocampus
(Fig. 3C, d, sector 8), whereas the populations of
cells with other pathological features were increased
exclusively in the cortex (Fig. 3C, b, c, e). Morphological and functional characteristics
of cortical and hippocampal neurons after
immunization against different p75NTR
fragments Interest-
ingly, the beneficial influence of the immunization
against 167–176 p75NTR fragment on neuronal mor-
phology in OBX-mice were observed, to a lesser
extent, in SO-mice as well (c.f., Fig. 3B and C, a,
b, d, sector 9). (For simplicity, the data associated
with Fig. 3 has been included in the Supplementary
materials.) of cortical and hippocampal neurons (Fig. 3B). This
immunological approach significantly improved their
morphological and functional characteristics, i.e.,
increased the number of healthy neurons (Fig. 3B,
a) at the expense of those with cytolysis, karyoly-
sis, and pyknosis (Fig. 3B, b, d, and e, respectively). The beneficial effect of the immunization against
p75NTR fragment of 167–176, expressed in the neu-
ronal density (Fig. 3B, f), in particular, was evident
in OBX- versus SO-mice (c.f., Fig. 3B, f versus Ñ,
f, sector 9, both). In contrast, enlarged population of
the vacuolated neurons (Fig. 3B, c) and decreased
neuronal density in OBX-mice treated with 32–46,
115–122, and 147–154 p75NTR fragments (Fig. 3B,
f, sectors 1, 6, and 7, respectively) was observed. Thus, protective effects of the immunization against
p75NTR fragments in OBX-mice were expressed
to a variable extent depending on the specificity of
the fragments; however, a common non-specific ten-
dency for some morphological characteristics was
revealed as well. To define this more exactly, an
additional series of experiments on 49 SO-mice was
performed with immunization against most of the p75
NTR fragments used on OBX-mice: 39–47; 86–93;
97–105; 147–154; 155–164, and 167–176. With the
exception of 167–176 p75NTR fragment (Fig. 3C,
sector 9), the immunization against other fragments
negatively affected the neuronal morphology in SO-
mice, supporting the conclusion that this treatment
specifically protects OBX-damaged brains from the
pathology seemingly associated with enhanced A
level. Maximal negative changes in SO-mice were Whether the beneficial effects of the immunization
on neuronal morphology is correlated with spatial
memory improvement, as tested in Morris water
maze, will be analyzed in the next section. Effects of the immunization against p75NTR
fragments on spatial memory In the beginning of the main experiments, individ-
ual swimming capabilities of mice were evaluated N. Bobkova et al. / p-75NTR specificity and cholinergic mediation in AD 296 Fig. 3. Morphological and functional characteristics of neurons in the temporal cortex and in the hippocampal areas of CA1 and CA3 after
immunization against different p75NTR fragments. A) Statistical characterization of non-immunized SO-and OBX-mice, where stars denote
significant (p < 0.01, Mann-Whitney U-test) difference between them. B) Radial presentation of the effects produced by the immunization
against each of studied fragments of p75NTR in OBX-mice. C) The same in SO-mice. A description to the radial diagrams is shown on I. (Supporting experimental data in numerical format is presented in Supplementary Tables 1 and 2). Fig. 3. Morphological and functional characteristics of neurons in the temporal cortex and in the hippocampal areas of CA1 and CA3 after
immunization against different p75NTR fragments. A) Statistical characterization of non-immunized SO-and OBX-mice, where stars denote
significant (p < 0.01, Mann-Whitney U-test) difference between them. B) Radial presentation of the effects produced by the immunization
against each of studied fragments of p75NTR in OBX-mice. C) The same in SO-mice. A description to the radial diagrams is shown on I. (Supporting experimental data in numerical format is presented in Supplementary Tables 1 and 2). in the water. Mean latencies to reach the visible
platform for OBX- and SO-mice (n = 82 and 49,
respectively) were approximately equal (7.3 ± 0.4 s
and 8.1 ± 0.6 s, respectively; t-test, p > 0.05) regard-
less of whether they were immunized or not. In the
following training days with the hidden platform, sig-
nificant decrease of latencies in the 4th versus the
1st session was observed in both OBX- and SO-mice
(Fig. 4A and 5A, two-way ANOVA: F1,140 = 104 and
F1,74 = 71, respectively, p ≤0.001, for both). Thus,
the animals from each group have learned to reach the
platform with reasonably latencies. OBX-mice were
distributed within the groups as follows (the number
of mice): Without (16) and after the immunization
with synthetic peptides, the analogs of the p75NTR
fragments of 32–46 (5), 39–47 (6), 66–72 (7), 86–93
(6), 97–105 (7), 115–122 (6), 147–154 (4), 155–164
(7), 167–176 (7), KLH (6), and VP1 (5). Effects of the immunization against p75NTR
fragments on spatial memory SO-mice did the same as follows: Without the immunization (15)
and after the immunization with the p75NTR frag-
ments of 39–47 (5), 86–93 (3), 97–105 (5), 147–154
(3), 155–164 (4), 167–176 (5), KLH (5), and VP1 (4). the same as follows: Without the immunization (15)
and after the immunization with the p75NTR frag-
ments of 39–47 (5), 86–93 (3), 97–105 (5), 147–154
(3), 155–164 (4), 167–176 (5), KLH (5), and VP1 (4). The memory testing revealed that the trained OBX-
mice from all groups spent significantly less time in
the target sector III than SO-mice (one-way ANOVA:
F12,83 = 8.9, p < 0.001), however, OBX-mice immu-
nized against 32–46, 155–164, and 167–176 p75NTR
fragments demonstrated partial memory improve-
ment versus non-immunized OBX-animals (Fig. 4B). The immunization against these fragments has been
shownabovetoberesponsibleforthebeneficialeffect
on the functioning of acetylcholine-synthesized neu-
rons in the basal forebrain (see Fig. 1A). Furthermore,
the treatment with 155–164 and 167–176 p75NTR
fragments evidently rescued morphology of cortical
and hippocampal neurons (see Fig. 3, a, b, d, e, sectors The memory testing revealed that the trained OBX-
mice from all groups spent significantly less time in
the target sector III than SO-mice (one-way ANOVA:
F12,83 = 8.9, p < 0.001), however, OBX-mice immu-
nized against 32–46, 155–164, and 167–176 p75NTR
fragments demonstrated partial memory improve-
ment versus non-immunized OBX-animals (Fig. 4B). The immunization against these fragments has been
shownabovetoberesponsibleforthebeneficialeffect
on the functioning of acetylcholine-synthesized neu-
rons in the basal forebrain (see Fig. 1A). Furthermore,
the treatment with 155–164 and 167–176 p75NTR
fragments evidently rescued morphology of cortical
and hippocampal neurons (see Fig. 3, a, b, d, e, sectors N. Bobkova et al. / p-75NTR specificity and cholinergic mediation in AD
297
Fig. 4. Learning (A) and memory (B) abilities of OBX-mice immunized against different fragments of p75NTR. A) Decreased latency in
reaching the saving platform on the 4th day (gray bars) versus that on the 1st day (white bars) of learning is shown. Stars denote significant
latency differences between the days: One and two markers are p < 0.05 and 0.01, respectively (Mann-Whitney U-test). B) In session with
memory testing, relative (versus “SO+V” control) time spending in the sector where the saving platform was placed during learning was
decreased in OBX-mice, with exception of partial recovery for those immunized against p75NTR fragments of 32–46, 155–164, and 167–176. Effects of the immunization against p75NTR
fragments on spatial memory A) Decreased latency in
reaching the saving platform on the 4th day (gray bars) versus that on the 1st day (white bars) of learning is shown. Stars denote significant
latency differences between the days: One and two markers are p < 0.05 and 0.01, respectively (Mann-Whitney U-test). B) In session with
memory testing, relative (versus “SO+V” control) time spending in the sector where the saving platform was placed during learning was
decreased in OBX-mice, with exception of partial recovery for those immunized against p75NTR fragments of 32–46, 155–164, and 167–176. Stars and pluses denote significant differences (one and two markers are p < 0.05 and 0.01, respectively, Mann-Whitney U-test) versus SO
and OBX, respectively. A, B) V, KLH, and VP1 denote Vehicle, Keyhole Limpet Hemocyanin conjugate, and a protein from foot-and-mouth
disease virus, respectively; error bars show 1 SEM. The only exception is associated with 167–176 frag-
ment whose immunological blockade produced the
cholinergic and morphological protective effects both
in OBX- and, partially, in SO-mice. 8 and 9). In contrast, spatial memory in SO-mice was
insensitive (one-way ANOVA: F7,37 = 1.9, p > 0.05)
to the immunization against those fragments which
were effective for OBX-mice (c.f., Figs. 4B and 5B). Moreover, the treatment with 86–93 and 147–154
p75NTR fragments tended to impair the memory in
SO-mice, and this was well correlated with the nega-
tive effects on neuronal morphology in the cortex and
hippocampus (see Fig. 3C, a - d, sectors 4 and 7). Effects of the immunization against p75NTR
fragments on spatial memory Stars and pluses denote significant differences (one and two markers are p < 0.05 and 0.01, respectively, Mann-Whitney U-test) versus SO
and OBX, respectively. A, B) V, KLH, and VP1 denote Vehicle, Keyhole Limpet Hemocyanin conjugate, and a protein from foot-and-mouth
disease virus, respectively; error bars show 1 SEM. N. Bobkova et al. / p-75NTR specificity and cholinergic mediation in AD 297 Fig. 4. Learning (A) and memory (B) abilities of OBX-mice immunized against different fragments of p75NTR. A) Decreased
reaching the saving platform on the 4th day (gray bars) versus that on the 1st day (white bars) of learning is shown. Stars denote s
latency differences between the days: One and two markers are p < 0.05 and 0.01, respectively (Mann-Whitney U-test). B) In se
memory testing, relative (versus “SO+V” control) time spending in the sector where the saving platform was placed during lea
decreased in OBX-mice, with exception of partial recovery for those immunized against p75NTR fragments of 32–46, 155–164, and
Stars and pluses denote significant differences (one and two markers are p < 0.05 and 0.01, respectively, Mann-Whitney U-test)
and OBX, respectively. A, B) V, KLH, and VP1 denote Vehicle, Keyhole Limpet Hemocyanin conjugate, and a protein from foot-a
disease virus, respectively; error bars show 1 SEM. Fig. 4. Learning (A) and memory (B) abilities of OBX-mice immunized against different fragments of p75NTR. A) Decreased latency in
reaching the saving platform on the 4th day (gray bars) versus that on the 1st day (white bars) of learning is shown. Stars denote significant
latency differences between the days: One and two markers are p < 0.05 and 0.01, respectively (Mann-Whitney U-test). B) In session with
memory testing, relative (versus “SO+V” control) time spending in the sector where the saving platform was placed during learning was
decreased in OBX-mice, with exception of partial recovery for those immunized against p75NTR fragments of 32–46, 155–164, and 167–176. Stars and pluses denote significant differences (one and two markers are p < 0.05 and 0.01, respectively, Mann-Whitney U-test) versus SO
and OBX, respectively. A, B) V, KLH, and VP1 denote Vehicle, Keyhole Limpet Hemocyanin conjugate, and a protein from foot-and-mouth
disease virus, respectively; error bars show 1 SEM. Fig. 4. Learning (A) and memory (B) abilities of OBX-mice immunized against different fragments of p75NTR. DISCUSSION Inthisstudy,theeffectsoftheimmunizationagainst
several selected p75NTR fragments in OBX-mice,
characterized by typical hallmarks of Alzheimer-type
neurodegeneration, were analyzed to reveal the most
effective path to develop a new perspective approach
forADtreatment.Wesuggestthattheobservedeffects Therefore, it is reasonable to conclude that, in
OBX- versus SO-mice, identical p75NTR fragments
are differently involved in different ways in memory
functioning and neuronal organization in the brain. 298 N. Bobkova et al. / p-75NTR specificity and cholinergic mediation in AD Fig. 5. Learning (A) and memory (B) abilities of SO-mice immunized against different fragments of p75NTR. A) Decreased latency in
reaching the saving platform on the 4th day (gray bars) versus that on the 1st day (white bars) of learning is shown. Stars denote significant
(p < 0.01, Mann-Whitney U-test) latency differences between the days. B) In session with memory testing, relative (versus “Vehicle” control)
time spending in the target sector was comparable in all groups of mice. A,B) KLH denotes Keyhole Limpet Hemocyanin conjugate; error g. 5. Learning (A) and memory (B) abilities of SO-mice immunized against different fragments of p75NTR. A) Decreased latency in
aching the saving platform on the 4th day (gray bars) versus that on the 1st day (white bars) of learning is shown. Stars denote significant
< 0.01, Mann-Whitney U-test) latency differences between the days. B) In session with memory testing, relative (versus “Vehicle” control)
me spending in the target sector was comparable in all groups of mice. A,B) KLH denotes Keyhole Limpet Hemocyanin conjugate; error
rs show 1 SEM. 5 L
i
(A)
d
(B) biliti
f SO
i
i
i
d
i
t diff
t f
t
f
75NTR A) D
d l t Fig. 5. Learning (A) and memory (B) abilities of SO-mice immunized against different fragments of p75NTR. A) Decreased latency in
reaching the saving platform on the 4th day (gray bars) versus that on the 1st day (white bars) of learning is shown. Stars denote significant
(p < 0.01, Mann-Whitney U-test) latency differences between the days. B) In session with memory testing, relative (versus “Vehicle” control)
time spending in the target sector was comparable in all groups of mice. A,B) KLH denotes Keyhole Limpet Hemocyanin conjugate; error
bars show 1 SEM. (Fig. 3B, f) that seemingly was associated with dis-
turbances in the apoptotic or, alternatively, autophagy
mechanism, noticed at the AD progression [30]. DISCUSSION The
increased density of ChAT-immunopositive neurons
in the forebrain observed in our study after immu-
nization against some p75NTR fragments (Fig. 1A)
is in line with evidence both of enhanced cholinergic
innervations in the cortex and hippocampus in knock-
out-mice (ChAT-cre p75(in/in) lacking p75NTR [31]
and increased numbers of such neurons [32]. In gen-
eral, our results are in line with evidence of the
critical role of p75NTR in the degeneration of cholin-
ergic neurons evoked by exposure either to A [13,
18], or to pro-/neurotrophins [33]. Indeed, the anti-
bodies against the extracellular p75NTR have been of the immunization were predominantly initiated by
a blockade of the fragment-associated binding sites in
the extracellular domain of p75NTR by appropriate
antibodies whose titers were sufficiently high in our
study (Table 1). The immunization against some of p75NTR frag-
ments in OBX-mice improved their spatial memory,
lowered cholinergic deficit and number of neurons
with apoptosis in the basal forebrain, and normalized
cellular morphology in the cortex and hippocam-
pus. Maximal positive effects were observed after the
immunization against 155–164 and 167–176 amino
acid residues of p75NTR. One negative consequence
of this treatment became apparent in the decrease
of neuronal density, especially, in the hippocampus N. Bobkova et al. / p-75NTR specificity and cholinergic mediation in AD 299 both memory and neuronal morphology, including
increased number of acetylcholine-producing cells,
in the forebrain were evident. In contrast, the treat-
ment with 86–93 and 147–154 p75NTR fragments
impaired the memory and neuronal characteristics in
SO-mice without significant negative influences on
OBX-mice. Interestingly, the immunization against
p75NTR167–176fragmentwasaccompaniedbypos-
itiveeffectsinbothOBX-andSO-mice.Thestructural
basis for these may include various conformational
transformations after the blockade of neurotrophin
or proneurotrophin binding to p75NTR. In this case,
conformational mechanisms apparently have to be
sensitive to the extracellular domain structures of the
receptor. It is well known that dimeric p75NTR is able
tobindtoNT-4receptorwithlowaffinityandtostimu-
lateneurogenesis.Moreover,p75NTR,inconjunction
with dimeric TrkA, TrkB, and TrkÑ receptors, has
shown to enhance efficacy of their binding with neu-
rotrophicfactorssuchasNGF,BDNF,NT-3,andNT-4
[38] to enhance, in turn, the survival rate of cells. shown to inhibit A-induced cell death and signif-
icantly diminished cell loss [20]. Furthermore, the
immunization against some p75NTR fragments in
OBX-mice was accompanied in our study by dimin-
ished numbers of apoptotic neurons in the cholinergic
areas of their forebrains (Fig. 2). DISCUSSION Moreover, enhanced
expression of both p75NTR, its major ligand, pro-
neurotrophin, and A, in AD brains [15, 34], suggest
that antibodies against binding sites for the ligands,
inducing apoptotic processes, will be able to improve
memory in OBX-mice and neuronal morphology in
their brains. We have revealed that the most protective effect
in OBX-mice was produced by the immunologi-
cal blockade of 155–164 and 167–176 fragments of
p75NTR. However, in spite of practically identical
roles of these amino acid residues for survival/death
of ChAT-containing neurons (Fig. 1A), the protective
effects of their blockade may originate from different
sources. We have shown recently that the immuno-
logical blockade of 155–164 residues in p75NTR of
OBX-mice, characterized by enhanced level of A
[29], was accompanied by significant decrease of
the brain A level, whereas immunization against
the 167–176 residue was ineffective [35]. Thus,
the 155–164 fragment of the p75NTR extracellular
domain may specifically be involved in the mech-
anisms of either A binding with p75NTR or A
aggregation [36], or, alternatively, in guiding A to
lysosomes for consecutive degradation in cholinergic
neurons [11]. Regardless of the mechanisms asso-
ciated with the lowering of A in the brain, the
immunological blockade of the 155–164 residue may
be useful approach for the brain protection from AD-
linked cholinergic deficit and memory impairment,
in particular, by the improving of the memory con-
solidation provided by synapse remodeling [37]. In conclusion, a study of interactive functioning
of p75NTR and Trk receptors by using of immuno-
logical means may be a key approach for both the
understanding of mechanisms of neuronal degener-
ation and the developing of effective new treatment
applicable for AD [13], aging [39] and other neu-
rodegenerative diseases associated with cholinergic
system malfunctioning in the brain. CONCLUSION In olfactory bulbectomized rats with typical hall-
marks of Alzheimer-type of degeneration, we have
demonstrated protective effects of immunization
against two of nine specific fragments of p75 neu-
rotrophin receptor extracellular domain on both
cholinergic neurons in the forebrain and cognitive
abilities in the animals. In summary, in spite of some differences in the
effects of the immunization against various p75NTR
fragments in OBX-mice, evident similarities were
observed in morphological characteristics of cortical
and hippocampal neurons (Fig. 3B), that might be
consideredasanon-specificactionoftheactiveimmu-
nization. However, this approach applied to SO-mice
showed either no or opposite effects. This suggests
that the pathological/neurodegenerative processes in
OBX-mice are accompanied by changes in the role
of the studied fragments of p75NTR extracellular
domain in cell survival. Thus, the immunological
blockade of p75NTR 155–164 fragment exerts either
a negligible or negative influence on neuronal char-
acteristics in SO-mice, whereas improvements of REFERENCES [17]
Blochl A, Blochl R (2007) A cell-biological model of
p75NTR signaling. J Neurochem 102, 289-305. [1]
Auld DS, Kornecook TJ, Bastianetto S, Quirion R (2002)
Alzheimer’s disease and the basal forebrain cholinergic
system: Relations to -amyloid peptides, cognition, and
treatment strategies. Prog Neurobiol 68, 209-245. [18]
Sotthibundhu A, Sykes AM, Fox B, Underwood CK, Thang-
nipon W, Coulson EJ (2008) Beta-amyloid (1-42) induces
neuronal death through the p75 neurotrophin receptor. J Neurosci 28, 3941-3946. [2]
Davis KL, Mohs RC, Marin D, Purohit DP, Perl DP, Lantz
M, Austin G, Haroutunian V (1999) Cholinergic markers
in elderly patients with early signs of Alzheimer disease. JAMA 281, 1401-1406. [19]
Coulson EJ, May LM, Sykes AM, Hamlin AS (2009) The
roleofthep75neurotrophinreceptorincholinergicdysfunc-
tion in Alzheimer’s disease. Neuroscientist 15, 317-323. [3]
DeKosky ST, Ikonomovic MD, Styren SD, Beckett L,
Wisniewski S, Bennett DA, Cochran EJ, Kordower JH,
Mufson EJ (2002) Upregulation of choline acetyltrans-
ferase activity in hippocampus and frontal cortex of elderly
subjects with mild cognitive impairment. Ann Neurol 51,
145-155. [20]
Hu Y, Lee X, Shao Z, Apicco D, Huang G, Gong BJ,
Pepinsky RB, Mi S (2013) A DR6/p75(NTR) complex is
responsible for -amyloid-induced cortical neuron death. Cell Death Dis 4, e579. [21]
Roux PP, Barker PA (2002) Neurotrophin signaling through
the p75 neurotrophin receptor. Prog Neurobiol 67, 203-233. [4]
Bobkova N, Vorobyov V (2015) The brain compensatory
mechanisms and Alzheimer’s disease progression: A new
protective strategy. Neural Regen Res 10, 696-697. [22]
Salehi A, Ocampo M, Verhaagen J, Swaab DF (2000) p75
neurotrophin receptor in the nucleus basalis of meynert in
relation to age, sex, and Alzheimer’s disease. Exp Neurol
161, 245-258. [5]
Nesterova IV, Bobkova NV, Medvinskaya NI, Samokhin
AN, Alexandrova IY (2008) Morphofunctional state of neu-
rons in the temporal cortex and hippocampus in relation
to the level of spatial memory in rats after ablation of the
olfactory bulbs. Neurosci Behav Physiol 38, 349-353. [23]
Reichardt LF (2006) Neurotrophin-regulated signalling
pathways. Philos Trans R Soc Lond B Biol Sci 361, 1545-
1564. [24]
He XL, Garcia KC (2004) Structure of nerve growth fac-
tor complexed with the shared neurotrophin receptor p75. Science 304, 870-875. [6]
Bobkova N, Guzhova I, Margulis B, Nesterova I, Medvin-
skaya N, Samokhin A, Alexandrova I, Garbuz D, Nudler E,
Evgen’ev M (2013) Dynamics of endogenous Hsp70 syn-
thesis in the brain of olfactory bulbectomized mice. Cell
Stress Chaperones 18, 109-118. REFERENCES [25]
Kamynina AV, Volpina OM, Medvinskaya NI, Alek-
sandrova IJ, Volkova TD, Koroev DO, Samokhin AN,
Nesterova IV, Shelukhina IV, Kryukova EV, Tsetlin VI,
Ivanov VT, Bobkova NV (2010) Vaccination with pep-
tide 173-193 of acetylcholine receptor 7-subunit prevents
memory loss in olfactory bulbectomized mice. J Alzheimers
Dis 21, 249-261. [7]
Coulson EJ (2006) Does the p75 neurotrophin receptor
mediate Abeta-induced toxicity in Alzheimer’s disease? J Neurochem 98, 654-660. [8]
Hempstead BL (2002) The many faces of p75NTR. Curr
Opin Neurobiol 12, 260-267. [26]
Bobkova NV, Medvinskaya NI, Kamynina AV, Aleksan-
drova IY, Nesterova IV, Samokhin AN, Koroev DO, Filatova
MP, Nekrasov PV, Abramov AY, Leonov SV, Volpina OM
(2014) Immunization against either prion protein fragment
95-123 or the fragment-specific antibodies rescue mem-
ory loss and neurodegenerative phenotype of neurons in
olfactory bulbectomized mice. Neurobiol Learn Mem 107,
50-64. [9]
Knowles JK, Rajadas J, Nguyen TV, Yang T, LeMieux MC,
Vander Griend L, Ishikawa C, Massa SM, Wyss-Coray T,
Longo FM (2009) The p75 neurotrophin receptor promotes
amyloid-beta(1-42)-induced neuritic dystrophy in vitro and
in vivo. J Neurosci 29, 10627-10637. [10]
Zhou XF, Wang YJ (2011) The p75NTR extracellular
domain: A potential molecule regulating the solubility and
removal of amyloid-. Prion 5, 161-163. [27]
Franklin KBJ, Paxinos G (2007) The Mouse Brain in Stereo-
taxic Coordinates, 3rd Edition, AP, Elsevier, NY. [11]
Ovsepian SV, Antyborzec I, O’Leary VB, Zaborszky L,
Herms J, Oliver Dolly J (2014) Neurotrophin receptor p75
mediates the uptake of the amyloid beta (A) peptide,
guiding it to lysosomes for degradation in basal forebrain
cholinergic neurons. Brain Struct Funct 219, 1527-1541. [28]
Unger JW, Schmidt Y (1993) Galanin-immunoreactivity
in the nucleus basalis of Meynert in the rat: Age-related
changes and differential response to lesion-induced cholin-
ergic cell loss. Neurosci Lett 153, 140-143. [12]
Zeng F, Lu JJ, Zhou XF, Wang YJ (2011) Roles of p75NTR
in the pathogenesis of Alzheimer’s disease: A novel thera-
peutic target. Biochem Pharmacol 82, 1500-1509. [29]
Aleksandrova IY, Kuvichkin VV, Kashparov IA, Medvin-
skaya NI, Nesterova IV, Lunin SM, Samokhin AN, Bobkova
NV (2004) Increased level of beta-amyloid in the brain of
bulbectomized mice. Biochemistry (Mosc) 69, 176-180. [13]
Mufson EJ, Counts SE, Perez SE, Ginsberg SD (2008)
Cholinergic system during the progression of Alzheimer’s
disease: Therapeutic implications. Expert Rev Neurother 8,
1703-1718. [30]
Peric A, Annaert W (2015) Early etiology of Alzheimer’s
disease: Tipping the balance toward autophagy or endoso-
mal dysfunction? ACKNOWLEDGMENTS This work was supported by a grant of Russian
Found of Basic Research (N.B.): KOMFI 13-04-
40106-H, entitled “Structure-functional studies of
p-75 receptor – molecular target for neurodegener-
ative diseases immunotherapy”. Authors’ disclosures available online (http://j-alz. com/manuscript-disclosures/16-0146r2). N. Bobkova et al. / p-75NTR specificity and cholinergic mediation in AD 300 SUPPLEMENTARY MATERIAL [15]
Chakravarthy B, M´enard M, Ito S, Gaudet C, Dal Pr`a I,
Armato U, Whitfield J (2012) Hippocampal membrane-
associated p75NTR levels are increased in Alzheimer’s
disease. J Alzheimers Dis 30, 675-684. The supplementary material is available in the
electronic version of this article: http://dx.doi.org/
10.3233/JAD-160146. [16]
Knowles JK, Simmons DA, Nguyen TV, Vander Griend
L, Xie Y, Zhang H, Yang T, Pollak J, Chang T, Arancio
O, Buckwalter MS, Wyss-Coray T, Massa SM, Longo FM
(2013) Small molecule p75NTR ligand prevents cognitive
deficits and neurite degeneration in an Alzheimer’s mouse
model. Neurobiol Aging 34, 2052-2063. REFERENCES Acta Neuropathol 129, 363-381. [14]
Mufson EJ, Kordower JH (1992) Cortical neurons express
nerve growth factor receptors in advanced age and
Alzheimer disease. Proc Natl Acad Sci U S A 89, 569-573. [31]
Boskovic Z, Alfonsi F, Rumballe BA, Fonseka S, Windels
F, Coulson EJ (2014) The role of p75NTR in cholinergic N. Bobkova et al. / p-75NTR specificity and cholinergic mediation in AD 301 (2011) p75NTR regulates Abeta deposition by increasing
Abeta production but inhibiting Abeta aggregation with its
extracellular domain. J Neurosci 31, 2292-2304. basalforebrainstructureandfunction.JNeurosci34,13033-
13038. basalforebrainstructureandfunction.JNeurosci34,13033-
13038. [32]
Greferath U, Trieu J, Barrett GL (2012) The p75
neurotrophin receptor has nonapoptotic antineurotrophic
actions in the basal forebrain. J Neurosci Res 90, 278-287. [37] [37]
Freir DB, Fedriani R, Scully D, Smith IM, Selkoe DJ,
Walsh DM, Regan CM (2011) A oligomers inhibit synapse
remodelling necessary for memory consolidation. Neuro-
biol Aging 32, 2211-2218. [33]
Friedman WJ (2000) Neurotrophins induce death of hip-
pocampal neurons via the p75 receptor. J Neurosci 20,
6340-6346. [38]
Skeldal S, Matusica D, Nykjaer A, Coulson EJ (2011) Pro-
teolytic processing of the p75 neurotrophin receptor: A
prerequisite for signalling? Neuronal life, growth and death
signalling are crucially regulated by intra-membrane prote-
olysis and trafficking of p75NTR. Bioessays 33, 614-625. [34]
Volosin M, Song W, Almeida RD, Kaplan DR, Hempstead
BL, Friedman WJ (2006) Interaction of survival and death
signaling in basal forebrain neurons: Roles of neurotrophins
and proneurotrophins. J Neurosci 26, 7756-7766. [35]
Vol’pina OM, Medvinskaia NI, Kamynina AV, Zaporozh-
skaia IaV, Aleksandrova IIu, Koroev DO, Samokhin
AN, Volkova TD, Arsen’ev AS, Bobkova NV (2014)
Immunization against a synthetic fragment 155-164 of neu-
rotrophin receptor p75 prevents memory loss and decreases
beta-amyloid level in mice with experimentally induced
Alzheimer’s disease [Russian]. Bioorg Khim 40, 451-457. [39]
Costantini C, Scrable H, Puglielli L (2006) An aging path-
way controls the TrkA to p75NTR receptor switch and
amyloid -peptide generation. EMBO J 25, 1997-2006. [40]
Rosen GD, Williams AG, Capra JA, Connolly MT, Cruz
B, Lu L, Airey DC, Kulkarni K, Williams RW (2000) The
Mouse Brain Library @ www.mbl.org. Int Mouse Genome
Conference 14:166. www.mbl.org. [36]
Wang YJ, Wang X, Lu JJ, Li QX, Gao CY, Liu XH, Sun Y,
Yang M, Lim Y, Evin G, Zhong JH, Masters C, Zhou XF
|
https://openalex.org/W2254063782
|
https://bmcbioinformatics.biomedcentral.com/counter/pdf/10.1186/1471-2105-16-S19-S1
|
English
| null |
On the distribution of cycles and paths in multichromosomal breakpoint graphs and the expected value of rearrangement distance
|
BMC bioinformatics
| 2,015
|
cc-by
| 8,648
|
© 2015 Feijão et al. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://
creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the
original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/
zero/1.0/) applies to the data made available in this article, unless otherwise stated. RESEARCH Open Access Open Access Pedro Feijão1,2*, Fábio Viduani Martinez1,2,3, Annelyse Thévenin1,2 Pedro Feijão1,2*, Fábio Viduani Martinez1,2,3, Annelyse Thévenin1,2 From Brazilian Symposium on Bioinformatics 2014
Belo Horizonte, Brazil. 28-30 October 2015 From Brazilian Symposium on Bioinformatics 2014
Belo Horizonte, Brazil. 28-30 October 2015 From Brazilian Symposium on Bioinformatics 2014
Belo Horizonte, Brazil. 28-30 October 2015 Abstract Finding the smallest sequence of operations to transform one genome into another is an important problem in
comparative genomics. The breakpoint graph is a discrete structure that has proven to be effective in solving
distance problems, and the number of cycles in a cycle decomposition of this graph is one of the remarkable
parameters to help in the solution of related problems. For a fixed k, the number of linear unichromosomal
genomes (signed or unsigned) with n elements such that the induced breakpoint graphs have k disjoint cycles,
known as the Hultman number, has been already determined. In this work we extend these results to
multichromosomal genomes, providing formulas to compute the number of multichromosal genomes having a
fixed number of cycles and/or paths. We obtain an explicit formula for circular multichromosomal genomes and
recurrences for general multichromosomal genomes, and discuss how these series can be used to calculate the
distribution and expected value of the rearrangement distance between random genomes. * Correspondence: pfeijao@cebitec.uni-bielefeld.de
1Faculty of Technology, Bielefeld University, Universitätsstraße 25, 33615
Bielefeld, Germany
Full list of author information is available at the end of the article Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 RESEARCH Background Input genomes
are A = {(1 2 3 4 5 6 7 8 9)} (black edges) and B = {(6 -1 4 5 -2),
(∘-9 3 8 7 ∘)} (dotted edges). We obtain an explicit formula for circular genomes and
recurrences for more general cases. Our paper is organized as follows. In the Preliminaries
section we give some definitions and notations. The
results for circular and general multichromosomal
genomes are presented in the next section, called The
Multichromosomal Hultman Number. The following
section presents some discussion about the distribution
of the rearrangement distance, derived from the multi-
chromosomal Hultman numbers, and the Conclusion
section presents final remarks and perspectives. The multichromosomal Hultman number
In this section, we extend the results of [3,4] for multi-
chromosomal genomes. There are two new aspects that
must be considered. First, since the breakpoint graph can
be decomposed in cycles and paths, we may have to
count not only cycles, but also paths. The other question
is about the identity genome. In the unichromosomal
case, the identity genome is easily defined. In the multi-
chromosomal case, it is not obvious which given genome
is the identity. When working on multichromosomal cir-
cular genomes, the identity is defined as in the unichro-
mosomal case. In the general case, working on genomes
with linear and circular chromosomes, we analyze two
types of identities for genomes: one with only one set of
circular chromosomes and another with a set of circular
chromosomes and a set of linear chromosomes. Preliminaries We represent multichromosomal genomes using a simi-
lar notation as in [5]. A gene is a fragment of DNA on
one of the two DNA strands in a chromosome, showing
its orientation. A gene is represented by an integer and
its orientation by a sign. The orientation of a gene g
allows us to distinguish its two extremities, the tail (gt)
and the head (gh). A chromosome is represented by a
sequence of genes, flanked in the extremities by telo-
meres (∘) if the chromosome is linear; otherwise, it is
circular. Genomes are represented as sets of chromo-
somes. An adjacency in a genome is either a pair of
consecutive gene extremities in a chromosome, or a
gene extremity adjacent to a telomere (a telomeric adja-
cency ). For instance, A = {(∘1 2 3 4 ∘)} is a genome
with one linear chromosome and four genes, and has
the adjacencies ∘1t, 1h2t, 2h3t, 3h4t and 4h∘, where the
first and the last are telomeric adjacencies. In the next sections, we propose extensions of the
Hultman number for multichromosomal genomes, first
considering only circular genomes, and then extending
the results to general genomes, with linear and circular
chromosomes. The same strategy is used in all cases:
first, start with a matching representing the identity, and
then superimpose all other possible matchings, while
counting recursively cycles and paths. To do that, we
need to consider all possible operations to build such
matchings. In Figure 4, all such operations are shown. There is a one-to-one correspondence between genomes
and matchings in the set of extremities. Adjacencies corre-
spond to two matched (saturated) vertices, and telomeric
adjacencies correspond to unmatched (unsaturated) ver-
tices. Therefore, a perfect matching (i.e., matching which
saturates all vertices of the graph) corresponds to a gen-
ome with only circular chromosomes. The matching cor-
responding to a genome A is denoted by MA. Because of
this one-to-one relationship, in this text we use the terms
genome and matching interchangeably. Background A remarkable characteristic of methods to compute
distances is the systematic use of a graph, first intro-
duced by Bafna and Pevzner [2], known as the break-
point graph. It has proven, by its decomposition into
disjoint cycles, a useful tool to efficiently compute rear-
rangement distances such as transposition or reversal,
directly related to the number of cycles in this decom-
position [1]. In molecular biology and genetics, comparative genomics
is a discipline interested in the comparison of genomic
attributes of different organisms. These attributes may
encompass DNA sequences, gene content, gene order,
regulatory sequences, and other structural features. Several measures have been proposed to compute the
(dis)similarity between genomes. The field called genome
rearrangements is concerned with measures of dissimilar-
ity involving large-scale mutations, such as reversals and
transpositions, where a fundamental problem is to deter-
mine the smallest sequence of such rearrangement
operations that transforms one given genome into
another. This minimum number of operations is called
the rearrangement distance between the two given gen-
omes. These and other aspects of genome rearrange-
ments are discussed in detail by Fertin et al. [1]. Since cycle decomposition of breakpoint graphs plays a
central role in computing distances, it is useful to investi-
gate the distribution of such cycles. Particularly, the dis-
tribution of genomes with a number of cycles c allows us
to evaluate the probability to have a scenario of a dis-
tance d depending of c. Doignon and Labarre [3] enum-
erated the unsigned permutations of a given size such
that the corresponding graph has a given number of
cycles, and called it the Hultman number. Subsequently,
Grusea and Labarre [4] extended this result for signed
permutations, where the signs model gene orientation. In this work we extend previous results providing for-
mulas to compute the number of multichromosomal
genomes with a given number of cycles and/or paths. * Correspondence: pfeijao@cebitec.uni-bielefeld.de
1Faculty of Technology, Bielefeld University, Universitätsstraße 25, 33615
Bielefeld, Germany
Full list of author information is available at the end of the article Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 Page 2 of 8 Figure 1 Multichromosomal breakpoint graph. Input genomes
are A = {(1 2 3 4 5 6 7 8 9)} (black edges) and B = {(6 -1 4 5 -2),
(∘-9 3 8 7 ∘)} (dotted edges). Figure 1 Multichromosomal breakpoint graph. General multichromosomal genomes HC(n, c) =
⎧
⎪⎪⎨
⎪⎪⎩
0,
if n = 0 and c > 0,
0,
if n < 0 or c < 0,
1,
if n = c = 0,
HC(n −1, c −1) + (2n −2) · HC(n −1, c), if n > c. The following result states an explicit formula to HC
(n, c). Theorem 1 Theorem 1
HC(n, c) =
2n−c
(c −1)!
0≤q1,...,qn−c:
n−c
2
mqm=n−c
(n + Q −1)! q2! · · · qn−c!1!q12!q2 . . . k!qn−c
,
where Q = q2 + ... + qk and
n−c
2
mqm = n −c is a
sum over all partitions of n - c. Figure 2 Construction of the breakpoint graph for a circular
identity genome I and a circular genome A. The adjacencies of I
are represented by black edges and those of A by grey edges. Unvisited nodes are white, visited ones are black. To build a perfect
match (circular genome A) only two operations are possible: (a)
Create a cycle; (b) Merge two paths. Theorem 1
HC(n, c) =
2n−c
(c −1)!
0≤q1,...,qn−c:
n−c
2
mqm=n−c
(n + Q −1)! q2! · · · qn−c!1!q12!q2 . . . k!qn−c
,
where Q = q2 + ... + qk and
n−c
2
mqm = n −c is a
sum over all partitions of n - c. HC(n, c) =
2n−c
(c −1)!
0≤q1,...,qn−c:
n−c
2
mqm=n−c
(n + Q −1)! q2! · · · qn−c!1!q12!q2 . . . k!qn−c
,
where Q = q2 + ... + qk and
n−c
2
mqm = n −c is a
sum over all partitions of n - c. Moreover, since we now have an operation that
is applied on just one vertex, and not two at a time
such as the operations presented in Section, we need
to define a different recurrence, where n correspond
to vertices in the breakpoint graph, and not to genes
in the genomes. In a genome I∘with n genes, there are
2n vertices (extremities) in MI◦and consequently in BG
(A, I∘). General multichromosomal genomes We will generalize our previous formula for general
multichromosomal genomes, with both linear and circu-
lar genomes. As already mentioned, two difficulties
arise. Now, we have not only cycles but also paths in
the breakpoint graph. Thus, it is not clear which gen-
ome should be considered the identity genome. As a
starting point, let us consider again the identity as I∘=
{(1, 2,..., n)}, and find the general Hultman number HG
(n, c, p), defined as (b) Merge Paths: If u and v belong to different com-
ponents, uv will merge both paths. There are 2n - 2 pos-
sibilities of adding such an edge. Applying any of the two operations results in a graph
with n - 1 paths, a subcase of the original graph with n
paths, with operation (a) also creating a cycle. This allows
us to establish a recurrence for HC (n, c). For the base
cases, when n = 0 we only have the empty genome, with
0 cycles in the breakpoint graph. Therefore, HC (0, c) = 1
if and only if c = 0, with HC (0, c) = 0 for c > 0. Also, if
either n or c is less than zero, we have that HC (n, c) = 0. (2) HG(n, c, p) ≡|{A ∈Gn : cyc(BG(A, I◦)) = c and pt(BG(A, I◦)) = p}|, HG(n, c, p) ≡|{A ∈Gn : cyc(BG(A, I◦)) = c and pt(BG(A, I◦)) = p}|,
(2 HG(n, c, p) ≡|{A ∈Gn : cyc(BG(A, I◦)) = c and pt(BG(A, I◦)) = p}|, where Gn is the set of all multichromosomal genomes
with n genes, and pt(·) denotes the number of paths in a
graph. In this set, each genome corresponds to a match-
ing, not necessarily perfect, since only circular genomes
correspond to perfect matchings. Similarly as the pre-
vious case, we start with the matching MI◦on 2n ver-
tices, and recursively build all possible matchings, while
counting cycles and paths. Since a matching induced by
an arbitrary genome A in Gn is not necessarily perfect,
together with the create cycle and merge paths opera-
tions on a vertex u, we can also choose to not saturate a
vertex u in the matching being built, thus creating a tel-
omere, which we call a skip vertex operation. explicit formula for result. □ Furthermore, the sequence of integers generated by
HC (n, c) is the unsigned entry A039683 in the OEIS
(On-Line Encyclopedia of Integer Sequences) [8]. (a) Create Cycle: If u and v belong to the same com-
ponent, the edge e = (u, v) will create a cycle. There is
only one possibility for this type of edge. Multichromosomal circular genomes g
A circular genome is a genome where all chromosomes
are circular. Since there are no telomeric adjacencies, the
matching MA of a circular genome A is a perfect match-
ing on the extremities of A. Moreover, the breakpoint
graph of two circular genomes is decomposed in disjoint
alternating cycles, since each vertex has degree two. Given two genomes A and B with the same set of
genes, the multichromosomal breakpoint graph of A and
B, denoted by BG(A, B), is built by joining the match-
ings MA and MB in the same set of vertices, using differ-
ent colors for the edges of each matching. Figure 1
shows an example of a multichromosomal breakpoint
graph for genomes A = {(1 2 3 4 5 6 7 8 9)} and B = {(6
-1 4 5 -2), (∘-9 3 8 7)}. From this point on we will use
the term breakpoint graph to refer to the multichromo-
somal breakpoint graph. Since all its vertices have
degree 0, 1 or 2, the breakpoint graph is uniquely
decomposed in cycles and paths. For instance, the
breakpoint graph in Figure 1 is decomposed in two
cycles and one path. We want to compute the number of circular genomes
with n genes that have c disjoint alternating cycles over a
given identity genome I, that we call the multichromoso-
mal circular Hultman number, denoted by HC (n, c). In
this case, since the matching of any circular genome is a
perfect matching, we claim that HC (n, c) is the same,
independently of the genome I chosen as an identity, and
simply define I∘= {(1, 2,..., n)}. Hence, we define (1) HC(n, c) ≡|{A ∈Cn : cyc(BG(A, I◦)) = c}|, Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 Page 3 of 8 Page 3 of 8 Proof We know from [6] that unsigned Stirling num-
bers of first kind satisfy the following recurrence equa- where Cn is the set of all circular multichromosomal
genomes with n genes and cyc(G) denotes the number
of cycles in a graph G. y
g
q
tion:
n
c
=
n −1
c −1
+ (n −1)
n −1
c
. Multiplying
both sides by 2n-c and using HC (n, c) recurrence equa-
tion we arrive at HC(n, c) = 2n−c
n
c
. Multichromosomal circular genomes Then, using the
explicit formula for
n
c
given in [7], we arrive at our
result. □ y
g
q
tion:
n
c
=
n −1
c −1
+ (n −1)
n −1
c
. Multiplying
both sides by 2n-c and using HC (n, c) recurrence equa-
tion we arrive at HC(n, c) = 2n−c
n
c
. Then, using the Starting with a perfect matching MI◦of the 2n ver-
tices, we build all breakpoint graphs BG(A, I∘), for circu-
lar genomes A, which correspond to perfect matchings,
adding one edge at a time, while counting the number
of cycles, recursively. The matching MI◦is composed by n connected com-
ponents, and all are paths. Considering an arbitrary ver-
tex u in the matching MI◦, there are 2n - 1 possible
edges uv that can be created. Figure 2 shows how these
different edges can be chosen. There are two possible
cases: Multichromosomal genomes with a fixed number of
linear chromosomes (a) Create Cycle: There is one edge uv such that v(≠u)
is the unvisited vertex in the same component as u, and
this edge (shown as a grey edge uv) will create a cycle. Vertices u and v are marked as visited (Figure 3(a)). In this section we generalize the previous approach for dif-
ferent identity genomes. Instead of fixing the identity as a
circular genome, the identity Iℓis a genome with a fixed
number of ℓlinear chromosomes. As for the input gen-
omes, first we consider all possible genomes, and in a sec-
ond approach also fix the number of linear chromosomes. Identity genome Iℓwith ℓlinear chromosomes
In this case, we can define the Hultman number (b) Merge Paths: There are e - 2 edges uv such that v
is an unvisited vertex in a different component as u, and
this edge will merge these components, that are paths. Vertices u and v are marked as visited. (Figure 3(b)). (c) Skip Vertex: Vertex u is not saturated; no edge is
created and only u is marked as visited (Figure 3(c)). (b) Merge Paths: There are e - 2 edges uv such that v
is an unvisited vertex in a different component as u, and
this edge will merge these components, that are paths. Vertices u and v are marked as visited. (Figure 3(b)). Identity genome Iℓwith ℓlinear chromosomes
In this case, we can define the Hultman number Vertices u and v are marked as visited. (Figure 3(b)). (c) Skip Vertex: Vertex u is not saturated; no edge is
created and only u is marked as visited (Figure 3(c)). ( g
( ))
(c) Skip Vertex: Vertex u is not saturated; no edge is
created and only u is marked as visited (Figure 3(c)). (3) HL(n, c, p, ℓ) ≡|{A ∈Gn : cyc(BG(A, Iℓ)) = c and pt(BG(A, Iℓ)) = p},
(3) HL(n, c, p, ℓ) ≡|{A ∈Gn : cyc(BG(A, Iℓ)) = c and pt(BG(A, Iℓ)) = p},
(3) If e is odd, it means that there is a vertex u that is
connected to a visited vertex. For this vertex, there is no
way to close a cycle, but the other two operations are
possible: where Gn is the set of all multichromosomal gen-
omes with n genes, and Iℓis a genome with exactly ℓ
linear chromosomes. General multichromosomal genomes So, we need an auxiliary number H′
G(e, c, p),
such that HG(n, c, p) = H′
G(e, c, p), with e = 2n, and
H′
G(e, c, p) ≡|{M ∈Me : cyc(BG(M, MI◦)) = c and pt(BG(M, MI◦)) = p}|,
where Me is the set of all possible matchings on e ver-
tices, and MI◦is a perfect matching with e/2 edges
induced by I∘. Figure 2 Construction of the breakpoint graph for a circular
identity genome I and a circular genome A. The adjacencies of I
are represented by black edges and those of A by grey edges. Unvisited nodes are white, visited ones are black. To build a perfect
match (circular genome A) only two operations are possible: (a)
Create a cycle; (b) Merge two paths. Figure 2 Construction of the breakpoint graph for a circular
identity genome I and a circular genome A. The adjacencies of I
are represented by black edges and those of A by grey edges. Unvisited nodes are white, visited ones are black. To build a perfect
match (circular genome A) only two operations are possible: (a)
Create a cycle; (b) Merge two paths. Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 Page 4 of 8 with (1) if any of e, c, p is negative, or e = 0 and any
of c, p is positive; (2) if e = c = p = 0; (3) if e is even;
and (4) if e is odd. Starting with the matching MI◦, another matching is
built recursively by adding edges or skipping vertices
until all vertices have been visited. Visited vertices are
shown in figures as black vertices, and unvisited as
white. If e is even, we pick any unvisited vertex u and
we have tree possibilities (Figure 3a-c): Multichromosomal genomes with a fixed number of
linear chromosomes This is a generalization of
the previous case, since HG(n, c, p) = HL(n, c, p, 0). We propose again an auxiliary series, defined as
H′
L(e, c, p, i) ≡|{M ∈Mn : cyc(BG(M, MIi)) = c and pt(BG(M, MIi)) = p}|,
where Mn is the set of all possible matchings on e
vertices, and MIi is a matching on these vertices such
that exactly i vertices are unsaturated (isolated), with
e = 2n and i = 2ℓ. Then, given a matching MIi with i
unsaturated vertices, we will build a matching recur-
sively adding edges or skipping vertices until all ver-
tices have been visited. In this case, the parity of e + i
determines which possibilities we have (Figure 4). When e + i is even, we will call the current state
balanced, otherwise it is unbalanced. In the balanced
case, focusing on an unvisited vertex u that is saturated
by MIi there are four possible cases (Figure 4a-d): (d) Merge Paths: There are e - 1 edges uv such that v is
in a different component as u, merging these components. Vertices u and v are marked as visited (Figure 3(d)). (d) Merge Paths: There are e - 1 edges uv such that v is
in a different component as u, merging these components. Vertices u and v are marked as visited (Figure 3(d)). (e) Skip Vertex: Vertex u is not saturated; no edge is
created, only u is marked as visited. A path where all
vertices are visited is created (Figure 3(e)). For the base cases, again we know that when e = 0, we
have only the empty genome, and this means that
H′
G(0, c, p) = 1
if
an
only
if
c
=
p
=
0,
and
H′
G(0, c, p) = 0 if c > 0 or p > 0. Also, if any of e, c, or
p is negative, H′
G(e, c, p) = 0. Multichromosomal genomes with a fixed number of
linear chromosomes With that, we arrive at
the following recurrence: H′
G(e, c, p) =
⎧
⎪⎪⎨
⎪⎪⎩
0,
(1)
1,
(2)
H′G(e −2, c −1, p) + (n −2) · H′G(e −2, c, p) + H′G(e −1, c, p), (3)
(n −1) · H′G(e −2, c, p) + H′G(e −1, c, p −1),
(4) y
i
g
(a) Create Cycle: There is one edge uv such that v
(≠u) is an unvisited vertex in the same component as u, (a) Create Cycle: There is one edge uv such that v
(≠u) is an unvisited vertex in the same component as u, Figure 3 Construction of the breakpoint graph for a circular genome I and a general genome A. The adjacencies of I are represented by
black edges and those of A by grey edges. Unvisited nodes are white, visited ones are black. We can create a cycle only when e (the number of
unvisited nodes) is even (a). We can merge two paths when e is even (b) or odd (d). We can skip a vertex when e is even (c) or odd (e). In (c)
and (d), the parity of the number of unvisited vertices is changed. Figure 3 Construction of the breakpoint graph for a circular genome I and a general genome A. The adjacencies of I are represented by
black edges and those of A by grey edges. Unvisited nodes are white, visited ones are black. We can create a cycle only when e (the number of
unvisited nodes) is even (a). We can merge two paths when e is even (b) or odd (d). We can skip a vertex when e is even (c) or odd (e). In (c)
and (d), the parity of the number of unvisited vertices is changed. Figure 3 Construction of the breakpoint graph for a circular genome I and a general genome A. The adjacencies of I are represented by
black edges and those of A by grey edges. Unvisited nodes are white, visited ones are black. We can create a cycle only when e (the number of
unvisited nodes) is even (a). We can merge two paths when e is even (b) or odd (d). We can skip a vertex when e is even (c) or odd (e). In (c)
and (d), the parity of the number of unvisited vertices is changed. Multichromosomal genomes with a fixed number of
linear chromosomes We can connect an unsaturated vertex when e = i
(h), when e + i is even (c) or odd (f). We can skip a vertex when e = i (i), when e + i is even (d) or when e + i odd (g). In (c) and (d), the parity
of e + i is changed. Figure 4 Construction of matching for genome Iℓwith ℓlinear chromosomes (i unsaturated vertices) and a general genome A. Adjacencies of I are represented by black edges and those of A by grey edges. Visited (unvisited) vertices are black (white). We can create a
cycle only when e + i is even (a). We can merge two paths when e + i is even (b) or odd (e). We can connect an unsaturated vertex when e = i
(h), when e + i is even (c) or odd (f). We can skip a vertex when e = i (i), when e + i is even (d) or when e + i odd (g). In (c) and (d), the parity
of e + i is changed. Figure 4 Construction of matching for genome Iℓwith ℓlinear chromosomes (i unsaturated vertices) and a general genome A. Adjacencies of I are represented by black edges and those of A by grey edges. Visited (unvisited) vertices are black (white). We can create a
cycle only when e + i is even (a). We can merge two paths when e + i is even (b) or odd (e). We can connect an unsaturated vertex when e = i
(h), when e + i is even (c) or odd (f). We can skip a vertex when e = i (i), when e + i is even (d) or when e + i odd (g). In (c) and (d), the parity
of e + i is changed. and this edge will create a cycle. Vertices u and v are
marked as visited. (g) Connect with unsaturated: There are i possible
edges from u to an unsaturated vertex v in Ii. Vertices u
and v are marked as visited, and a path with all vertices
visited is created. (b) Merge Paths: There are e - 2 - i edges uv such
that v is saturated in Ii and is in a different component
as u, and uv will merge these components, that are
paths. Vertices u and v are marked as visited. Multichromosomal genomes with a fixed number of
linear chromosomes Cases (e), (f) and (g) are similar to cases (b), (c) and
(d), respectively, which means that (e) keeps the state
unbalanced, but (f) and (g) change it to balanced again. There are still two cases to consider, when e = i (Figure
4h,i). (c) Skip Vertex: No edge is created and u is marked as
visited. (c) Skip Vertex: No edge is created and u is marked as
visited. (d) Connect with unsaturated: There are i possible
edges from u to an unsaturated vertex v in Ii. Vertices u
and v are marked as visited. (d) Connect with unsaturated: There are i possible
edges from u to an unsaturated vertex v in Ii. Vertices u
and v are marked as visited. (h) Connect two unsaturated: There are i - 1 possible
edges from an unsaturated vertex u to an unsaturated
vertex v in Ii. Vertices u and v are marked as visited,
and a path with all vertices visited is created. Cases (a) and (b) visit two vertices that are saturated in
Ii, which means that the state remains balanced. Case (c)
changes the state to unbalanced, since only one vertex is
visited. Case (d) visits two vertices, but one is a unsatu-
rated vertex in Ii, which means that the parity of e + i
changes and the state becomes unbalanced. (i) Skip Vertex: No edge is created and u is marked
as visited. A path with all vertices visited is created. For the base cases, as before when e = 0 we have
H′
L(0, c, p, i) = 1 if and only if c = p = i = 0, and
H′
L(0, c, p, i) = 0 if any of c, p, i is positive. Also, if any
of e, c, p, i is negative, H′
L(e, c, p, i) = 0. In the unbalanced state, focusing on a vertex u
belonging to a component with all other vertices visited,
there are three possibilities (Figure 4e-g): With all these cases described, we arrive at the recur-
rence, from what we can deduce HL(n, c, p, ℓ): (e) Merge Paths: There are e - 1 - i edges uv such
that v is saturated in Ii and is in a different component
as u, and this edge will merge these components, that
are paths. Vertices u and v are marked as visited. Multichromosomal genomes with a fixed number of
linear chromosomes Figure 3 Construction of the breakpoint graph for a circular genome I and a general genome A. The adjacencies of I are represented by
black edges and those of A by grey edges. Unvisited nodes are white, visited ones are black. We can create a cycle only when e (the number of
unvisited nodes) is even (a). We can merge two paths when e is even (b) or odd (d). We can skip a vertex when e is even (c) or odd (e). In (c)
and (d), the parity of the number of unvisited vertices is changed. Figure 3 Construction of the breakpoint graph for a circular genome I and a general genome A. The adjacencies of I are represented by
black edges and those of A by grey edges. Unvisited nodes are white, visited ones are black. We can create a cycle only when e (the number of
unvisited nodes) is even (a). We can merge two paths when e is even (b) or odd (d). We can skip a vertex when e is even (c) or odd (e). In (c)
and (d), the parity of the number of unvisited vertices is changed. Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 Page 5 of 8 Figure 4 Construction of matching for genome Iℓwith ℓlinear chromosomes (i unsaturated vertices) and a general genome A. Adjacencies of I are represented by black edges and those of A by grey edges. Visited (unvisited) vertices are black (white). We can create a
cycle only when e + i is even (a). We can merge two paths when e + i is even (b) or odd (e). We can connect an unsaturated vertex when e = i
(h), when e + i is even (c) or odd (f). We can skip a vertex when e = i (i), when e + i is even (d) or when e + i odd (g). In (c) and (d), the parity
of e + i is changed. Figure 4 Construction of matching for genome Iℓwith ℓlinear chromosomes (i unsaturated vertices) and a general genome A. Adjacencies of I are represented by black edges and those of A by grey edges. Visited (unvisited) vertices are black (white). We can create a
cycle only when e + i is even (a). We can merge two paths when e + i is even (b) or odd (e). Multichromosomal genomes with a fixed number of
linear chromosomes H′
L(e, c, p, i) =
⎧
⎪⎪⎪⎪⎪⎪⎪⎪⎪⎨
⎪⎪⎪⎪⎪⎪⎪⎪⎪⎩
0,
(1)
1,
(2)
(i −1) · H′L(e −2, c, p −1, i −2) + H′L(e −1, c, p −1, i −1),
H′L(e −2, c −1, p, i) + (e −2 −i) · H′L(e −2, c, p, i)+
(3)
i · H′L(e −2, c, p, i −1) + H′L(e −1, c, p, i),
(4)
(e −1 −i) · H′L(e −2, c, p, i)+
i · H′L(e −2, c, p −1, i −1) + H′L(e −1, c, p −1, i),
(5) H′
L(e, c, p, i) =
⎧
⎪⎪⎪⎪⎪⎪⎪⎪⎪⎨
⎪⎪⎪⎪⎪⎪⎪⎪⎪⎩
0,
(1)
1,
(2)
(i −1) · H′L(e −2, c, p −1, i −2) + H′L(e −1, c, p −1, i −1),
H′L(e −2, c −1, p, i) + (e −2 −i) · H′L(e −2, c, p, i)+
(3)
i · H′L(e −2, c, p, i −1) + H′L(e −1, c, p, i),
(4)
(e −1 −i) · H′L(e −2, c, p, i)+
i · H′L(e −2, c, p −1, i −1) + H′L(e −1, c, p −1, i),
(5) (f) Skip Vertex: Vertex u is not saturated in M ; no
edge is created and only u is marked as visited, and a
path with all vertices visited is created. i · H′L(e −2, c, p −1, i −1) + H′L(e −1, c, p −1, i),
(5) Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 Page 6 of 8 introduction in 2005, because it can model several rear-
rangement operations and it is commonly easy to calcu-
late in many cases. The DCJ distance between two
genomes A and B is given by d(A, B) = n - c - e/2,
where n is the number of genes, and c and e are respec-
tively the number of cycles and even paths (paths with
even number of edges) in the breakpoint graph BG(A,
B). Using group theory, an alternative measure called
algebraic rearrangement distance was proposed by Feijäo
and Meidanis [11]. This distance can also be calculated
with the breakpoint graph, namely da(A, B) = n - c -
p/2, where n is the number of genes, and c and p are
respectively the number of cycles and paths in the
breakpoint graph BG(A, B). Identity genome Iℓi and input genomes Aℓa with ℓi and ℓa
linear chromosomes For instance, if we have the random
variable Xn = da(An, In), where In is the circular identity
of size n and An is a genome sampled uniformly from
the set Cn of all circular genomes, then we have Therefore, the recurrence is given by H′
ℓ(e, c, p, i, a) =
⎧
⎪⎪⎪⎪⎪⎪⎪⎪⎪⎪⎪⎪⎨
⎪⎪⎪⎪⎪⎪⎪⎪⎪⎪⎪⎪⎩
0,
(1)
1,
(2)
(i −1) · H′ℓ(e −2, c, p −1, i −2, a)+
H′ℓ(e −1, c, p −1, i −1, a −1),
(3)
H′ℓ(e −2, c −1, p, i) + (e −2 −i, a) · H′ℓ(e −2, c, p, i, a)+
i · H′ℓ(e −2, c, p, i −1, a) + H′ℓ(e −1, c, p, i, a −1),
(4)
(e −1 −i) · H′ℓ(e −2, c, p, i, a)+
i · H′ℓ(e −2, c, p −1, i −1, a) + H′ℓ(e −1, c, p −1, i, a −1), (5) H′
ℓ(e, c, p, i, a) =
⎧
⎪⎪⎪⎪⎪⎪⎪⎪⎪⎪⎪⎪⎨
⎪⎪⎪⎪⎪⎪⎪⎪⎪⎪⎪⎪⎩
0,
(1)
1,
(2)
(i −1) · H′ℓ(e −2, c, p −1, i −2, a)+
H′ℓ(e −1, c, p −1, i −1, a −1),
(3)
H′ℓ(e −2, c −1, p, i) + (e −2 −i, a) · H′ℓ(e −2, c, p, i, a)+
i · H′ℓ(e −2, c, p, i −1, a) + H′ℓ(e −1, c, p, i, a −1),
(4)
(e −1 −i) · H′ℓ(e −2, c, p, i, a)+
i · H′ℓ(e −2, c, p −1, i −1, a) + H′ℓ(e −1, c, p −1, i, a −1), (5) P
Xn = d
= DC(n, d)
|Cn|
=
DC(n, d)
(2n −1)!!, since |Cn| is the number of circular genomes of size n
and corresponds to the number of perfect matchings
with 2n vertices, given by (2n - 1)!!. The expected value
is then given by with (1) if any of e, c, p, i is negative, or e = 0 and any of
c, p, i is positive; (2) if e = c = p = i = 0; (3) if e = i > 0, (4)
if e + i is even, e > i, (5) if e + i is odd, e > i. E [Xn] =
n
d=0
d · P
Xn = d
=
1
(2n −1)!! n
d=0
d · HC(n, n −d), Identity genome Iℓi and input genomes Aℓa with ℓi and ℓa
linear chromosomes DC(n, d) ≡|{A ∈Cn : da(A, I◦) = d}| = HC(n, n −d),
DG(n, d) ≡|{A ∈Gn : da(A, I◦) = d}| =
c+p/2=n−d
HG(n, c, p),
DL(n, d, ℓ) ≡|{A ∈Gn : da(A, Iℓ) = d}| =
c+p/2=n−d
HL(n, c, p, ℓ),
Dℓ(n, d, ℓi, ℓa) ≡|{Aℓa ∈Gn, ℓa : da(A, Iℓ) = d}| =
c+p/2=n−d
Hℓ(n, c, p, ℓi, ℓa). The base cases are also similar, only including a in the
constraints. When e = 0 we have H′
ℓ(0, c, p, i, a) = 1 if
and only if c = p = i = a = 0, and H′
ℓ(0, c, p, i, a) = 0
if any of c, p, i, a is positive. Also, if any of e, c, p, i, a is
negative, H′
ℓ(e, c, p, i, a) = 0. Using those equations, we can also calculate the
expected value for the rearrangement distance in any
selected scenario. Identity genome Iℓi and input genomes Aℓa with ℓi and ℓa
linear chromosomes In this scenario, in addiction to fixing ℓi linear chromo-
somes for the identity Iℓi, we also build breakpoint
graphs only with genomes Aℓa that have exactly ℓa linear
chromosomes. We propose the Hultman number (4) Hℓ(n, c, p, ℓi, ℓa) ≡|{Aℓa ∈Gn,ℓa : cyc(BG(A, Iℓ)) = c and pt(BG(A, Iℓ)) = p},
(4) were Gn,ℓa is the set of all multichromosomal gen-
omes with n genes and exactly ℓa linear chromosomes,
and Iℓis, as before, a genome with exactly ℓlinear
chromosomes. By
definition,
we
have
that
n
ℓa=0 Hℓ(n, c, p, ℓi, ℓa) = HL(n, c, p, ℓi). were Gn,ℓa is the set of all multichromosomal gen-
omes with n genes and exactly ℓa linear chromosomes,
and Iℓis, as before, a genome with exactly ℓlinear
chromosomes. By
definition,
we
have
that
n
ℓa=0 Hℓ(n, c, p, ℓi, ℓa) = HL(n, c, p, ℓi). Again we define an auxiliary series, in this case
H′
ℓ(e, c, p, i, a) ≡|{M ∈Me, a : cyc(BG(M, Me, i)) = c and pt(BG(M, Me, i)) =
p}|
,
where Me, a is the set of all possible matchings on e
vertices that has exactly a unsaturated vertices, and MIi
is a matching on these vertices such that exactly i ver-
tices are unsaturated. To build the breakpoint graph for
this new series, we use exactly the same operations as in
the previous, summarized in Figure 4. The only differ-
ence is that we have to track how many unsaturated
vertices a the current matching being build has. The
only operations that change this are the skip vertex
operations (c), (i) and (f), decreasing a by 1. The other
operations keep a the same, as they all create an edge
and do not mark any vertex as unsaturated. DC(n, d) ≡|{A ∈Cn : da(A, I◦) = d}| = HC(n, n −d),
DG(n, d) ≡|{A ∈Gn : da(A, I◦) = d}| =
c+p/2=n−d
HG(n, c, p),
DL(n, d, ℓ) ≡|{A ∈Gn : da(A, Iℓ) = d}| =
c+p/2=n−d
HL(n, c, p, ℓ),
Dℓ(n, d, ℓi, ℓa) ≡|{Aℓa ∈Gn, ℓa : da(A, Iℓ) = d}| =
c+p/2=n−d
Hℓ(n, c, p, ℓi, ℓa). Multichromosomal genomes with a fixed number of
linear chromosomes Since the parity of paths is
not important in the algebraic distance, it is the best
suited model for calculating the distribution of the rear-
rangement distances from the Hultman numbers pro-
posed here. For each of the four cases, we ask the
following question: How many genomes of size n have
distance d from a given identity genome? Making the
same assumptions about the identity and also the uni-
verse of the genomes - that is, circular only, general, or
a fixed number of linear chromosomes -, we arrive in
the following distance distributions, shown also in
Figure 5. It is interesting to notice that most of the gen-
omes are very distant from the identity. with (1) if any of e, c, p, i is negative, or e = 0 and any of
c, p, i is positive; (2) if e = c = p = i = 0; (3) if e = i > 0, (4)
if e + i is even, e > i, (5) if e + i is odd, e > i. with (1) if any of e, c, p, i is negative, or e = 0 and any of
c, p, i is positive; (2) if e = c = p = i = 0; (3) if e = i > 0, (4)
if e + i is even, e > i, (5) if e + i is odd, e > i. Distribution of rearrangement distances The number of general genomes of size n is the number of
matchings with 2n vertices, which is the telephone number
T (n) (sequence A000085 in OEIS [8]), given by
T(n) = ⌊n/2⌋
k=0
n! 2k(n −2k)!k! . The equations below follow:
n
c=0
HC(n, c) = (2n −1)!!,
n
c=0
n
p=0
HG(n, c, p) = T(n), Another interesting relationship is that, for a fixed n, the
sum of all combination of cycles and paths in a series results
in the number of genomes of size n. The number of circular
genomes of size n corresponds to the number of perfect
matchings with 2n vertices, which is given by (2n - 1)!!. The number of general genomes of size n is the number of
matchings with 2n vertices, which is the telephone number
T (n) (sequence A000085 in OEIS [8]), given by
⌊/ ⌋
n! Distribution of rearrangement distances From the Hultman series that we introduced, it is possi-
ble to derive the distribution of rearrangement distances
for each scenario. and can therefore be calculated with the given recur-
rence equations. For instance, for n = 100 we have E
[X100] = 95.22. A closed formula for the expected value of
a rearrangement distance, to the best of our knowledge, The Double Cut and Join (DCJ) distance [9,10] is one
of the most studied rearrangement distances since its Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 Page 7 of 8 Figure 5 Distribution of the rearrangement distance between genomes of size n = 21, in four different scenarios. Figure 5 Distribution of the rearrangement distance between genomes of size n = 21, in four different scenarios. has only been found for the very simple breakpoint dis-
tance dBP, which counts how many adjacent genes in the
identity are not adjacent in the other genome, and is For the Hultman number HC (n, c), in addition to the
recursive equations we also provided an explicit formula,
using the relationship between this series and the
unsigned Stirling numbers of first kind. An interesting
future direction is finding explicit formulas for the other
proposed sequences HG(n, c, p) and HL(n, c, p, ℓ). y
j
g
given by E[dBP(An, In)] = n −
1
2 + 1
2n + O
1
n2
[12]. y
j
g
given by E[dBP(An, In)] = n −
1
2 + 1
2n + O
1
n2
[12]. This converges to n - 1/2 when n goes to infinity, which
is almost the diameter n for the breakpoint distance. Although we have no closed formula for E[Xn], we con-
jecture that it also converges to n - k for some constant
k > 0, as n goes to infinity, and the experimental results
point to k ≈5. Another interesting relationship is that, for a fixed n, the
sum of all combination of cycles and paths in a series results
in the number of genomes of size n. The number of circular
genomes of size n corresponds to the number of perfect
matchings with 2n vertices, which is given by (2n - 1)!!. Authors’ contributions Authors’ contributions
All authors contributed equally. All authors contributed equally. Authors’ details
1 1Faculty of Technology, Bielefeld University, Universitätsstraße 25, 33615
Bielefeld, Germany. 2Institute for Bioinformatics, Center for Biotechnology
(CeBiTec), Bielefeld University, Bielefeld, Germany. 3Faculdade de
Computação, Universidade Federal de Mato Grosso do Sul, 79070-900
Campo Grande Brazil Published: 16 December 2015 Submit your next manuscript to BioMed Central
and take full advantage of:
• Convenient online submission
• Thorough peer review
• No space constraints or color figure charges
• Immediate publication on acceptance
• Inclusion in PubMed, CAS, Scopus and Google Scholar
• Research which is freely available for redistribution
Submit your manuscript at
www.biomedcentral.com/submit Declarations Publication fees were funded by Deutsche Forschungsgemeinschaft and the
OpenAccess Publication Funds of Bielefeld University. This article has been published as part of BMC Bioinformatics Volume 16
Supplement 19, 2015: Brazilian Symposium on Bioinformatics 2014. The full
contents of the supplement are available online at http://www. biomedcentral.com/bmcbioinformatics/supplements/16/S19 Acknowledgements FVM is funded from the Brazilian research agency CNPq grant Ciência sem
Fronteiras Postdoctoral Scholarship 245267/2012-3. We acknowledge support
of the publication fee by Deutsche Forschungsgemeinschaft and the Open
Access Publication Funds of Bielefeld University. Conclusions In this paper, we introduced different recursive formulas
for the Hultman number and its variations, that are
relevant in the context of comparative genomics. We
have extended previous results that treated the unichro-
mosomal cases [3,4], focusing on multichromosomal
genomes. Table 1 shows a summary of the results. T(n) = ⌊n/2⌋
k=0
n! 2k(n −2k)!k! . The equations below follow: n
c=0
HC(n, c) = (2n −1)!!,
n
c=0
n
p=0
HG(n, c, p) = T(n), Table 1 Summary of the results in this paper. Hultman Number
Identity
Universe
H(n, k) [3]
π = 〈... 〉
Sn (unsigned permutations)
H±(n, k) [4]
π = 〈... 〉
S±
n (signed permutations)
HC (n, c)
Circular genome
Circular genomes
HG(n, c, p)
Circular genome
General genomes
HL(n, c, p, ℓ)
Genome with ℓlinear chr. General genomes
Hℓ(n, c, ℓi, ℓa)
Genome with ℓi linear chr. Genomes with ℓa linear chr. The first two rows show previous results, and the last four show the Hultman numbers proposed in this paper. Table 1 Summary of the results in this paper. and the last four show the Hultman numbers proposed in this pap The first two rows show previous results, and the last four show the Hultman numbers proposed in this paper. Page 8 of 8 Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 Feijão et al. BMC Bioinformatics 2015, 16(Suppl 19):S1
http://www.biomedcentral.com/1471-2105/16/S19/S1 n
c=0
n
p=0
HL(n, c, p, ℓ) = T(n),
for ℓ= 0, . . . , n. and
n
ℓa=0
n
c=0
n
p=0
Hℓ(n, c, p, ℓi, ℓa) = T(n),
for ℓi = 0, . . . , n. 7. Malenfant J: Finite, closed-form expressions for the partition function
and for Euler, Bernoulli, and Stirling numbers., ArXiv e-prints (2011). 1103.1585. 8. Sloane NJA: The On-Line Encyclopedia of Integer Sequences - OEIS 2014
[http://oeis.org]. 9. Yancopoulos S, Attie O, Friedberg R: Efficient sorting of genomic
permutations by translocation, inversion and block interchange. Bioinformatics 2005, 21(16):3340-3346. 10. Bergeron A, Mixtacki J, Stoye J: A unifying view of genome
rearrangements. Lecture Notes in Computer Science 2006, 4175:163-173. 11. Feijao P, Meidanis J: Extending the algebraic formalism for genome
rearrangements to include linear chromosomes. IEEE/ACM Transactions on
Computational Biology and Bioinformatics 2013, 10(4):819-831, doi:10.1109/
TCBB.2012.161. 12. Xu W, Alain B, Sankoff D: Poisson adjacency distributions in genome
comparison: multichromosomal, circular, signed and unsigned cases. Conclusions Bioinformatics 2008, 24(16):146-152, doi:10.1093/bioinformatics/btn295. doi:10.1186/1471-2105-16-S19-S1
Cite this article as: Feijão et al.: On the distribution of cycles and paths
in multichromosomal breakpoint graphs and the expected value of
rearrangement distance. BMC Bioinformatics 2015 16(Suppl 19):S1. 7. Malenfant J: Finite, closed-form expressions for the partition function
and for Euler, Bernoulli, and Stirling numbers., ArXiv e-prints (2011). 1103.1585. 8. Sloane NJA: The On-Line Encyclopedia of Integer Sequences - OEIS 2014
[http://oeis.org]. 9. Yancopoulos S, Attie O, Friedberg R: Efficient sorting of genomic
permutations by translocation, inversion and block interchange. Bioinformatics 2005, 21(16):3340-3346. 10. Bergeron A, Mixtacki J, Stoye J: A unifying view of genome
rearrangements. Lecture Notes in Computer Science 2006, 4175:163-173. 11. Feijao P, Meidanis J: Extending the algebraic formalism for genome
rearrangements to include linear chromosomes. IEEE/ACM Transactions on
Computational Biology and Bioinformatics 2013, 10(4):819-831, doi:10.1109/
TCBB.2012.161. 12. Xu W, Alain B, Sankoff D: Poisson adjacency distributions in genome
comparison: multichromosomal, circular, signed and unsigned cases. Bioinformatics 2008, 24(16):146-152, doi:10.1093/bioinformatics/btn295. doi:10.1186/1471-2105-16-S19-S1
Cite this article as: Feijão et al.: On the distribution of cycles and paths
in multichromosomal breakpoint graphs and the expected value of
rearrangement distance. BMC Bioinformatics 2015 16(Suppl 19):S1. and These equations might be useful for finding explicit
equations for some of the numbers. We wrote a Python
script with all recurrence relations proposed, and the
above equations were useful to check the correctness of
each series. 11. Feijao P, Meidanis J: Extending the algebraic formalism for genome
rearrangements to include linear chromosomes. IEEE/ACM Transactions on
Computational Biology and Bioinformatics 2013, 10(4):819-831, doi:10.1109/
TCBB.2012.161. 12. Xu W, Alain B, Sankoff D: Poisson adjacency distributions in genome
comparison: multichromosomal, circular, signed and unsigned cases. Bioinformatics 2008, 24(16):146-152, doi:10.1093/bioinformatics/btn295. The Hultman number can also be used to find the
expected value of the rearrangement distance between
uniformly distributed genomes, in our case the algebraic
distance between multichromosomal genomes. Future
directions include finding explicit equations for the intro-
duced recursive equations and the expected value of the
rearrangement distance. doi:10.1186/1471-2105-16-S19-S1
Cite this article as: Feijão et al.: On the distribution of cycles and paths
in multichromosomal breakpoint graphs and the expected value of
rearrangement distance. BMC Bioinformatics 2015 16(Suppl 19):S1. Competing interests
The authors declare that they have no competing interests. Competing interests
The authors declare that they have no competing interests. Authors’ contributions
All authors contributed equally. References Submit your next manuscript to BioMed Central
and take full advantage of: Submit your next manuscript to BioMed Central
and take full advantage of: Submit your next manuscript to BioMed Central
and take full advantage of: References
1. Fertin G, Labarre A, Rusu I, Tannier E, Vialette S: Combinatorics of Genome
Rearrangements. MIT Press, Cambridge, MA; 2009. 2. Bafna V, Pevzner PA: Genome rearrangements and sorting by reversals. SIAM Journal on Computing 1996, 25(2):272-289. 3. Doignon J, Labarre A: On Hultman numbers. Journal of Integer Sequences
2007, 10(6), Article 07.6.2, 13 p.. 4. Grusea S, Labarre A: The distribution of cycles in breakpoint graphs of
signed permutations. Discrete Applied Mathematics 2013, 161(10-
11):1448-1466. 5. Bergeron A, Mixtacki J, Stoye J: A unifying view of genome
rearrangements. Lecture Notes in Computer Science 2006, 4175:163-173. 6. Graham RL, Knuth DE, Patashnik O: Concrete Mathematics: A Foundation
for Computer Science. Addison-Wesley, USA; 1994. 1. Fertin G, Labarre A, Rusu I, Tannier E, Vialette S: Combinatorics of Genome
Rearrangements. MIT Press, Cambridge, MA; 2009. 2. Bafna V, Pevzner PA: Genome rearrangements and sorting by reversals. SIAM Journal on Computing 1996, 25(2):272-289. • Convenient online submission 3. Doignon J, Labarre A: On Hultman numbers. Journal of Integer Sequences
2007, 10(6), Article 07.6.2, 13 p.. 4. Grusea S, Labarre A: The distribution of cycles in breakpoint graphs of
signed permutations. Discrete Applied Mathematics 2013, 161(10-
11):1448-1466. 5. Bergeron A, Mixtacki J, Stoye J: A unifying view of genome
rearrangements. Lecture Notes in Computer Science 2006, 4175:163-173. 6. Graham RL, Knuth DE, Patashnik O: Concrete Mathematics: A Foundation
for Computer Science. Addison-Wesley, USA; 1994.
|
https://openalex.org/W2074661576
|
https://zenodo.org/records/1617396/files/article.pdf
|
English
| null |
COOPERATIVE INVESTIGATIONS ON PLANTS
|
Biometrika
| 1,903
|
public-domain
| 15,640
|
Biometrika n * BiometTtka, Vol. i. pp. 11—20.
+ Ibid., pp. 125—128.
X Ibid., pp. 816—319. II.
VARIATION AND CORRELATION IN LESSEE
CELANDINE FROM DIVERS LOCALITIES. y
y ,
p
j
g Mr Galton's plan was carried out in the further collections made in Guernsey,
Dorset and Surrey, the collectors gathering the buds, and wrapping them up Biometrika n Biometrika n 19 Cooperative Investigations on Plants 140 in situ in a small piece of tissue paper. The huds dried and were preserved for
weeks, to be counted at leisure. Even if a petal or sepal fell off it was preserved
in the paper wrapper. We believe it is largely due to this method of gathering
that our results show such a totally different distribution of Bepals and petals to
those of other observers. Take the case of sepals. In the Guernsey series we, found only three in-
dividuals with less than three sepala
On examination under the microscope
in one of these cases an abortive sepal was found, in another the sepal had
clearly once been attached, but in the third case the head was unfortunately lost
before microscopic examination. We think that we may safely affirm, that no true
case of less than three sepals was found in the Guernsey plants. In the Dorset-
shire gathering no cases of less thau three sepals occurred. In the Surrey
gathering there were in the material six cases of less than three sepals, and
in five out of these six cases the rule of gathering buds only had been disregarded
by a young collector*; and the sixth was an aborted flower in which the stamens
were not properly developed and there were no pistils at all! In the Bordighera
celandines there are only two cases of less than three sepals. These have not
been excluded from the calculations, because we had not when counting them
learnt from the Dorset and Guernsey series to regard all cases of less than three
sepals with grave suspicion and examine such cases under the microscope. But if
in 624 cases only two such individuals occurred, it seems extremely probable that
even these were cases of sepals knocked off or aborted. In a total of 2149 heads,
there were 11 cases of less than three sepals, two of these were aborted flowers,
five of them were old flowers with parts loose, one had once had a sepal which had
been lost, and three were not closely examined. II.
VARIATION AND CORRELATION IN LESSEE
CELANDINE FROM DIVERS LOCALITIES. (1) IN view of the data for F. ranunculoides published by Dr F. Ludwig*
and Prof. MacLeod f and the statistical constants determined for themj it seemed
desirable to obtain rather more statistical material and a more comprehensive
series of constants for the purposes of a comparative study. There were two
points to be considered, namely: (i) the influence of locality and (ii) the influence
of the time during the flowering season at which the flowers were gathered. Unfortunately we were in the present season in no case able to obtain from our
collectors two series from the same locality with a month's interval between the
gatherings. All the collecting except in one case had to be done during the
brief Easter vacation of our workers, and this did not admit of a double gathering
in the same locality at a suitable interval. The one exception is that of the
Bordighera collection. Mr Francis Galton most kindly offered, as he was wintering
on the Mediterranean, to provide a double series of Lesser Celandine flowers. The first series was gathered about February 19th and at once dispatched to
England. On arrival it was found that practically every sepal, every petal, and
nearly every stamen had fallen from the flower-heads. This lost us our earlier
series, but we learnt a most valuable lesson, namely: that transit of any kind,
even by hand, will cost the flower if nearly full-blown one or more sepals, petals or
stamena
Our plans had therefore to be changed ; each celandine flower was
now gathered as a bud and wrapped up in a small piece of tissue paper. This
involved a great increase in the labour of gathering and a much greater one in
that of counting, a good deal of which had to be done under a lens, but we were
thus certain of preserving all the parts of the flower intact. Mr Francis Galton
suggested and carried out this arrangement in a second series gathered between
March 4th and 7th which reached England safely, but three or four weeks later
than this there were no flowering celandines to be obtained in Bordighera. * The c&lonlator'i notes are : No. 51, "only one sepal and this banging loose " ; No. 62, " old flower
with petals detached"; No. 53, "Pistils loose"; No. 64, " very old flower, all parts loose"; No. 69,
" very old flower." No remarks as to age of this kind occur in other cases.
t See Biometrika, Vol. i. pp. 18—15. II.
VARIATION AND CORRELATION IN LESSEE
CELANDINE FROM DIVERS LOCALITIES. The experience we have had
leads us to believe that each flower ought to be gathered as a bud and at once
wrapped up. It seems to us that all we can admit is a possibility of three heads
with less than three sepals in 2149 cases, while Dr Ludwig's 3000 from Greiz
show no less than 60 definite cases f. We feel fairly confident that had the
Greiz flowers been collected as buds and possibly gathered and counted by adults
instead of school children they would not have differed so widely from our material
in this respect. As far as our experience reaches, we question the existence
of any normal flower with less than three sepals. This view may be modified
when further material from central Europe gathered in bud and, if necessary,
microscopically examined, is available. We hope next season that this problem of
the sepal may be directly investigated. II.
Variation and Correlation in Lesser Celandine The counting is due to Miss Edna Lea-Smith and to
Mr Blancbard. The statistical tables and constants are again due to Dr Lee. In reducing the material Sheppard's corrections for the moments were not
used, partly because we are dealing with variation by units, and partly because
it is clear in the case of the sepals, and probably true for the petals, that the
frequency distribution has not high contact at the low end of the range. Other-
wise the calculation of means, standard deviations, coefficients of correlation, and
of variation proceeded in the usual manner. p (D) 505 heads gathered in a lane at Studland, Dorsetshire, on April 7 by
Mr N. L. Blanchard. The counting is due to Miss Edna Lea-Smith and to
Mr Blancbard. The statistical tables and constants are again due to Dr Lee. (D) 505 heads gathered in a lane at Studland, Dorsetshire, on April 7 by
Mr N. L. Blanchard. The counting is due to Miss Edna Lea-Smith and to
Mr Blancbard. The statistical tables and constants are again due to Dr Lee. In reducing the material Sheppard's corrections for the moments were not
used, partly because we are dealing with variation by units, and partly because
it is clear in the case of the sepals, and probably true for the petals, that the
frequency distribution has not high contact at the low end of the range. Other-
wise the calculation of means, standard deviations, coefficients of correlation, and
of variation proceeded in the usual manner. In reducing the material Sheppard's corrections for the moments were not
used, partly because we are dealing with variation by units, and partly because
it is clear in the case of the sepals, and probably true for the petals, that the
frequency distribution has not high contact at the low end of the range. Other-
wise the calculation of means, standard deviations, coefficients of correlation, and
of variation proceeded in the usual manner. p
j (3) Table L gives a summary of all" the statistical constants* for the four new
series and places alongside them those already found for other localities. Now
this table shows at once the remarkable differences, which period in season and
environment can have on mean, variability and correlation. (2) Material of tlie present investigation (A) 624 heads gathered in a vineyard at Bordighera, Italy, between March 4th
and 7th- Due to Mr Francis Galton. The sepals, petals, stamens and pistils
were counted by Mr N. Blanchard. The tables were prepared and the statistical (A) 624 heads gathered in a vineyard at Bordighera, Italy, between March 4th
and 7th- Due to Mr Francis Galton. The sepals, petals, stamens and pistils
were counted by Mr N. Blanchard. The tables were prepared and the statistical II.
Variation and Correlation in Lesser Celandine II. Variation and Correlation in Lesser Celandine 147 constants calculated by Miss Alice Lee. These belonged to a variety of Lesser
Celandine classed by the French botaniste as Ficaria calthaefolia and considered
by them as distinct from F. ranuncuioides. Table L, however, exhibits less
differentiation in the floral parts of F. calthaefolia and F. ranunculoides than can
occur between two local races of the latter. The Index Kewensis gives the former
name as a synonym of the latter, but there appears to be a more sensible differen-
tiation of the leaf. constants calculated by Miss Alice Lee. These belonged to a variety of Lesser
Celandine classed by the French botaniste as Ficaria calthaefolia and considered
by them as distinct from F. ranuncuioides. Table L, however, exhibits less
differentiation in the floral parts of F. calthaefolia and F. ranunculoides than can
occur between two local races of the latter. The Index Kewensis gives the former
name as a synonym of the latter, but there appears to be a more sensible differen-
tiation of the leaf. (B) 520 heads gathered on roadside banks at St Peter Port, Guernsey,
between April 25th and 30th. Due to Miss Caroline Herford. The characters
were counted, the tables formed and the statistical constants determined by
Miss Mary Beeton. (B) 520 heads gathered on roadside banks at St Peter Port, Guernsey,
between April 25th and 30th. Due to Miss Caroline Herford. The characters
were counted, the tables formed and the statistical constants determined by
Miss Mary Beeton. y
(C) 500 heads gathered in two lanes and a meadow near Thursley, on the
north side of Hind Head, Surrey. The material was gathered between 12th and
17th of April by a number of collectors under the supervision of K. Pearson. The characters were counted by Mr N. Blanchard and the tables and statistical
constants are due to Dr Lee. (C) 500 heads gathered in two lanes and a meadow near Thursley, on the
north side of Hind Head, Surrey. The material was gathered between 12th and
17th of April by a number of collectors under the supervision of K. Pearson. The characters were counted by Mr N. Blanchard and the tables and statistical
constants are due to Dr Lee. (D) 505 heads gathered in a lane at Studland, Dorsetshire, on April 7 by
Mr N. L. Blanchard. II.
Variation and Correlation in Lesser Celandine Not one of these
quantities has the least approximation to constancy for all local races of Ficaria,
nor for the same race at two parts of the same reason. The early Belgian
celandines, judging from the constants for stamens and pistils, which are all we
have, are practically identical with those from Italy, while the later ones diverge
widely and perhaps may be considered nearer to the Swiss Trogen series than to
any other. A brief study of this table will at once convince the reader of two
fundamental points:
at Carleton University on July
ournals.org/ (a) Local races in plants cannot be defined or distinguished by the existence
of differences many times the value of their, probable errors between their means,
variations or correlationa
1, 2015 (6) The influences of environment and season are for plants of supreme impor-
tance and very widely or indeed entirely screen any differences due to local race. (6) The influences of environment and season are for plants of supreme impor-
tance and very widely or indeed entirely screen any differences due to local race. * These are tabulated to four places of decimals—not because such are exact or necessary for present
purposes, but because for future investigations when we come to consider the evolutionary history of local
races, it will be needful to have the constants to this number of places in order to calculate the partial
regression coefficients true for each series to one or two places of decimals. * These are tabulated to four places of decimals—not because such are exact or necessary for present
purposes, but because for future investigations when we come to consider the evolutionary history of local
races, it will be needful to have the constants to this number of places in order to calculate the partial
regression coefficients true for each series to one or two places of decimals. 10—2 10—2 ABLE 1. Statistical
Constants of Lesset* Celan Year
Date
Number
SeiwJs
S. Petals
Stamens
Pistils
Sepals
"5 1 Petals
Stamens
Pistils
|
| (Sepals ... • S ^ l 1 Petals ... t6,°C 1 Stamens... <§ tS ( Pistils ... Sepals & Potals... ,5
•g
II
Sepals & Stamens
Sepals & Pistils... Petals & Stamens
Petals and Pistils... II.
Variation and Correlation in Lesser Celandine Stamens and Pistils
Italy
Bordighera
1902
March 4-7
624
3-0625
±0071
81186
±O117
26-7179
±•1018
17-7051
±•1068
•2612
±0060
•4334
±0083
3-7720
+ 0720
3-9568
±O765
8-63
5-34
1412
22-35
1044
±O267
0228
±0270
•0706
±O269
•2243
±0256
1961
+ O260
•4387
±O218
Guernsey
St Peter Port
1902
April 25-80
520
3-0096
±0029
81327
±O137
27-9558
±•1419
23-9115
±•1877
•0976
+ O020
•4638
±0097
4-7978
+ •1003
6-3470
±•1327
3-24
5-70
1716
26-54
•0993
±O293
•0584
±0295
•0324
+ O295
•2939
±0264
•2222
±O281
•5206
±O216
England
Surrey
1902
April 12-17
500
3O100
±0030
8-0680
±O139
32-9380
±•1721
24-5360
± -1931
•0995
±0021
-4598
±0098
57039
±•1217
64001
±•1366
331
570
17-32
2608
•2912
+ O276
•0222
±0300
1078
±O297
•2690
±0280
•3508
±O266
•6601
±0170
Dorset
1902
April 7-9
606
3-0297
±0054
8-7149
+ O304
35-5881
±•1803
28-3287
±•2242
1811
+ O038
1-0118
±O215
6-0058
±1276
7-4713
+ •1586
5 98
11-61
16-88
26-37
•2300
±O284
•2225
±O285
•2490
±O282
•3825
±0256
•3527
±O263
•6682
±O166
Belgium
Early
1899
Feb. 27—
March 17
268
26-7313
±•1660
17-4478
±•1604
—
3-7609
±•1096
3-8943
±•1135
1407
22-32
—
—
—
•5065
±O306
Late
1899
April 17-28
878
17-8633
±•1162
121475
±•1183
_
3 2984
±O815
33878
±0837
18-46
27-89
•7489
±0153
Germany
Greiz
1900-1901
4000
3-660
±•0171
8-351
±0260
—
•8037
±O121
1-2189
±O184
—
—
22-02
14-60
—
—
•2451
±0200
—
—
—
—
Gera*
1
1000
3-309
±O137
8-226
±0237
—
-6406
±0097
11113
±O168
—
19-36
13-51
—
1928
±O205
—
—
—
—
[-
±
* A second series from Gera while less variable gives sensibly the same means and correlation. A third series, more or less
an insensible negative correlation: see Biometrika, vol. i. p. 817. t. This result is very discordant and is not included in the European mean: Bee Biometrika, Vol. i. p. 317. j A seoond short series for Gain, with somewhat different means and variabilities, is fairly accordant in oorrelation. at Carleton University on July 1, 2015
http://biomet.oxfordjournals.org/
Downloaded from era*
1
000
309
O137
226
237
—
-6406
0097
1113
O168
—
36
-51
—
1928
O205
—
—
—
—
Tr
1
2
33
±0
81
±0
20-
±•
13-
±•
•
±0
•
±O
38
±•
3-
±O
16-
9-
18-
23-
[-•i
±03
±0
ore or less int
on. era*
1
1000
309
O137
226
0237
—
-6406
0097
1113
O168
—
-36
-51
—
1928
O205
—
—
—
—
Tro
19
2
33
±0
81
±0
20-
±•1
13-
±•1
•
±0
•
±O
38
±•1
3-
±O
16-
9-
18-
23-
[-•i8
±03
•
±0
ore or less int
on. II.
Variation and Correlation in Lesser Celandine 149 These results form a further proof, were any needful, that race cannot be
defined by correlation*. Plants are, of course, more subject to environment than
animal life, and the means and variabilities change with change of environment
and of season very readily. Man, perhaps, of all animal life seems least affected
by environment, but a table like the above showing how different the mean, varia-
bility and correlation are when there is slight change in locality or season, ought
to render us cautious of cataloguing even local races of man by the means alone
of small craniological series. Nurture and environment during growth may
sensibly influence even human mean charactere. The bioinetrician, far from being
discouraged by the fact that the simpler biometric quantities—means, variabilities,
correlations—are not constant for local races, ought rather to recognise that if
natural selection be a reality these are the very quantities which he would expect
to change from one local race to another, and from one environment to a second. Indeed it is exactly these differences which form the foundation upon which we
hope to build up in the future the evolutionary history of local races; they are
the material from which we must determine not only the characters which in
each case have been selected, but also the absolute constants which define the
species itselff. Downloaded from (4) Comparing the first four columns—the new data—with the last four
columns—the old data for central Europe—one is at once struck with the high
degree of variability in sepals and petals of the latter as compared with the
former. The sepals are more than four times, the petals are nearly twice, as
variable in the second group, ie. the mean values are "6544 and 1-0946 as agaiust
•1598 and "5944 respectively. Now we see in the case of the Dorset group that
the petal variability can rise to double the value it had in Bordighera, Guernsey
or Surrey. But an examination of the tables below Bhows that much of the
difference arises from the absence of individuals with less than 3 sepals or 7 petals
in the first group, and as our data are drawn from very diverse districts we must
await with interest the result of bud countings from Germany. ,
, pp
,
+ " Mathematical Contributions to the Theory of Evolution. XI. On the Influence of Natural
Selection on Variation and Correlation." Phil. Tram. Vol. 200, A, pp. 18—21. S
"5
|
• S
t6
<§
,5
•g
II
Pe
St
an II. Variation and Correlation in Lesser Celandine II. Variation and Correlation in Lesser Celandine * Phil. Tram. VoL 187, A, pp. 266, 280. II.
Variation and Correlation in Lesser Celandine p
j
g (4) Comparing the first four columns—the new data—with the last four
columns—the old data for central Europe—one is at once struck with the high
degree of variability in sepals and petals of the latter as compared with the
former. The sepals are more than four times, the petals are nearly twice, as
variable in the second group, ie. the mean values are "6544 and 1-0946 as agaiust
•1598 and "5944 respectively. Now we see in the case of the Dorset group that
the petal variability can rise to double the value it had in Bordighera, Guernsey
or Surrey. But an examination of the tables below Bhows that much of the
difference arises from the absence of individuals with less than 3 sepals or 7 petals
in the first group, and as our data are drawn from very diverse districts we must
await with interest the result of bud countings from Germany. at Carleto
http://biomet.oxfordjournals.org/ * Phil. Tram. VoL 187, A, pp. 266, 280. TABLE III. But making all allowance for this the German and Swiss series show ft marked
increase in the number of both sepals and petals over our series, culminating
iu Gais with its modes of 5 sepals and 10 petals. Our Dorsetshire series indicates
a substantial advance in the same direction as far as the petals are concerned, the
Bordighera tends somewhat in the same direction for the petals. It remains as a
task for next spring to determine whether the Guernsey and Gais series cannot be
linked together by a continuous system of intermediate series, if only the flowers
be gathered at a sufficiently wide range of places and at different seasons in those
places. http://biomet. Downloaded from (5) Turning to the correlations which involve sepals one is hardly surprised
at their irregularity, for their values depend in our new data on the distribution of
a very few individuals with more than four sepala These for Bordighera are
less than 7 per cent, for Dorset less than 3 per cent., and for Guernsey and Surrey
not more than 1 per cent. I Hence one or two irregular or anomalous individuals
cause the correlation to alter in a very remarkable manner. A much safer set
of correlations are those between petals, stamens and pistils. Unfortunately the
counting of stamens and pistils is a much harder task, and for a series of 500
bud-gatherings requires a week or two of very laborious work. at Carleton Unive
oxfordjournals.org/ (5) Turning to the correlations which involve sepals one is hardly surprised
at their irregularity, for their values depend in our new data on the distribution of
a very few individuals with more than four sepala These for Bordighera are
less than 7 per cent, for Dorset less than 3 per cent., and for Guernsey and Surrey
not more than 1 per cent. I Hence one or two irregular or anomalous individuals
cause the correlation to alter in a very remarkable manner. A much safer set
of correlations are those between petals, stamens and pistils. Unfortunately the
counting of stamens and pistils is a much harder task, and for a series of 500
bud-gatherings requires a week or two of very laborious work. TABLE I I TABLE I I
Number of Sepals
Bordighera..,
. St Peter Port
. Surrey
Dorset
Greiz
Gera
Trogen
... . Gais
Frequency of Sepals per
0
—
4-2
30
1
—
1-8
o-o
2
3-2
90
210
20
S
932-7
990-4
990-0
972-3
546-8
733-0
680-7
91-0
1000. 4
62-6
9-6
10-0
257
2517
162-0
259-7
283-0
5
1-6
2-0
174-2
86O
52-6
616O
0
—
11-5
5O
7O
7-0
—
0-8
1-0 Cooperative Investigations on Plants Cooperative Investigations on Plants Cooperative Investigations on Plants 150 TABLE III. Frequency of Petals per 1000. Number of Petals
Bordighem... St Peter Port
Surrey
Dorset
Greiz
Gera
Trogen
Gaw
• S
—
—
—
0-2
—
—
—
4
—
—
—
0-8
—
—
—
—
—
6-8
16O
3-5
—
0
1-6
—
4-0
—
32-7
31-0
14-0
2-0
7
14-4
135
40-0
7-9
148-0
112-0
119-3
34-0
8
868-6
869-2
868O
660-4
473-0
563-0
6421
1390
9
962
961
84O
231-7
180-0
175-0
160-9
265-0
10
17-6
13-6
10-0
124-7
99-0
62-0
69-7
296-0
11
1-6
7-7
4-0
63-4
&V3
24-0
10-5
175-0
IS
7-9
17-2
11-0
—
69-0
13
—
4-0
6-0
5-0
18-0
n
—
—
0-6
o-o
2-0
15
—
—
0-6
1-0
—
— TABLE III. Frequency of Petals per 1000. TABLE III. II.
Variation and Correlation in Lesser Celandine I. Variation and Correlation in Lesser Celandine 151 The slight exception to this rule—Surrey correlation of petals and stamens—is
well within the probable error of the determinations. Summing up these results in a single statement, we conclude from the data
that were gathered and counted on one uniform plan that: The local races of the lesser celandine which have flowers with more numerous
parts, have those parts more variable and more highly correlated. Let us compare these results with the data obtained by other observers. Germany contributes nothing, for stamens and pistils have not yet been counted. The Swiss data, if individually a little anomalous, form, if clubbed together, a
striking confirmation of our argument,—they have fewer pistils and stamens than
our four series, the variations are less, and the correlations for stamens and pistils
are low—Gais has the lowest result of all, aud Trogen only about the Guernsey
value. Unfortunately we can only compare one correlation and this has been
worked out for only small numbers, 285 in one case and 80 in the other. With
regard to Belgium the comparative results are not so satisfactory. The early
Belgium flowers have about the same number of stamens as the Italian and fewer
pistils, the variability of both stamens and pistils is less. The late Belgium
flowers have the least number of stamens and pistils of any series, their variability
in both stamens and pistils is less than all four of our series, and less than the
mean of the two Swiss series. Here again we have unfortunately only one corre-
lation available for comparison, that of stamens and pistils. It is for the early
flowers somewhat greater than that for the Italian series, when it ought to be
about equal to it, while for the late flowers, it is the highest of all when it should
be the least from the above rule. But just in this material a new factor comes
into play, which, if not the source of the result, deserves to be taken into account. The late flowers are said to be from the same plants as the early flowers. But
pistils and stamens cannot be counted unless the flower be gathered. TABLE III. at Carleton Univ
oxfordjournals.org/ Looking at our new data, which are arranged according to number of stamens,
we see several important and almost uniform results:
ersity on (a) If the different series be arranged in ascending order of stamens, this will
also be an ascending order for the number of pistils. July 1, 2 With one exception—Surrey—this is also an ascending order for the number
of petals. 2015 With one exception—Surrey—this is also an ascending order for the number
of petals. 2015 (b) The ascending order for stamens or pistils is also an ascending order
for the variability of both stamens and pistils. With the same exception—Surrey—it is an ascending order for variability of
petals. (c) The ascending order for .means or variabilities of stamens or pistils is an
ascending order for the three correlations between stamens, pistils and petals. II.
Variation and Correlation in Lesser Celandine Hence the
plants from which the later flowers are gathered maybe heterogeneous in character,
namely plants naturally flowering late, and plants which have been more or less
injured by one or more flowers being removed and which have reflowered in
a more or less exhausted condition. Such heterogeneity would tend to increase
correlation. Clearly in this as in so many cases the conditions are very complex,
and we cannot without further investigation club together flowers from plants
(a) which have' had earlier flowers gathered, (b) which naturally flower late, with
the late flowers of plants which have been flowering throughout the season
undisturbed, and term the group ' late flowers' without danger of introducing
heterogeneity. Till therefore the whole subject of the correlation of late and
early flowers has been gone into carefully with ample data, we may take, we hold,
the above general principle—that increase in the number of parts denotes increase
in the variability and correlation of those parts—as a working hypothesis to
be demonstrated or modified by further researches. We, of course, do not
extend the principle beyond the species for which the data are deduced, even as a
" working hypothesis," and we state it only for petals, stamens and pistils. * And therefore its probable error eren with only eight pairs, so Email. Cooperative Investigations on Plants 152 (6) It may be useful to examine the interracial result from the correlation
standpoint, although at present we have rather meagre data. We have eight local
series; considering them of equal weight, although the Belgian and Swiss series
are very inferior in number to ours, we find : Mean of local means for stamens
= 26498 ± 1321
Mean of local means for pistils
= 19-432 ± 1265
Variability of local means for Htamens
... = 55409 ± '9343
Variability of local means for pistils
... = 5'3037 ± 8943
Correlation of local means for stamens and pistils = '9514 ± 0235. The probable errors are very high because we have only eight pairs. It is there-
tore difficult, perhaps, to make a definite statement that the interracial is greater
than the average intraracial variability, but it appears like it; even the relative at Carleton University
http://biomet.oxfordjournals.org/
Downloaded from m
at Carleton University on July 1, 2015
http://biomet.oxfordjournals.org/
Downloaded from ao
u
M
. «
*O
to
a
^
•
7
FRRA
" Celandine. J
/ \
PtSh
f,
i
i
1 UF
L
>
1
Interracial
AM—
»TE
j
1
1ELO U M ,
\ / /
\
I
0>
ReQTeuion Linei
y
c
/ ,
>4RC
c
1AIS,
y
OEN
UERf
Stament and . 8EV(
/
1
_P"
1
I S
|i
•*
/
RDIO
RLY
HERA
IELQ
DOR
Putilt. ™A
//
/OSUBRE
UM
/
V
1
Number of Suimeni in Racial Mean. order of variability of pistils and stamens may be changed when we pass from
intraracial to interracial constants. On the other hand the correlation is so high *
that
e can make a
er
definite statement abo t it i e :
at Carleton University on July 1, 2015
http://biomet.oxfordjournals.org/
Downloaded from ao
u
M
. «
*O
to
a
^
•
7
FRRA
" Celandine. J
/ \
PtSh
f,
i
i
1 UF
L
>
1
Interracial
AM—
»TE
j
1
1ELO U M ,
\ / /
\
I
0>
ReQTeuion Linei
y
c
/ ,
>4RC
c
1AIS,
y
OEN
UERf
Stament and . 8EV(
/
1
_P"
1
I S
|i
•*
/
RDIO
RLY
HERA
IELQ
DOR
Putilt. ™A
//
/OSUBRE
UM
/
V
1
Number of Suimeni in Racial Mean. Cooperative Investigations on Plants order of variability of pistils and stamens may be changed when we pass from
intraracial to interracial constants. On the other hand the correlation is so high *
y
y ,
p
j
g 7
" Celandine. Interracial ReQTeuion Linei
Stament and .Putil at Carleton University on July 1, 2015
http://biomet.oxfordjournals.org/
from . *O
a
^ j
Number of Suimeni in Racial Mean. order of variability of pistils and stamens may be changed when we pass from
intraracial to interracial constants. On the other hand the correlation is so high *
that we can make a very definite statement about it, i.e.: order of variability of pistils and stamens may be changed when we pass from
intraracial to interracial constants. On the other hand the correlation is so high *
that we can make a very definite statement about it, i.e.: The interracial correlation of the mean numbers of stamens and pistils is very
much greater than the mean intraracial correlation between stamens and pistils,
being to the latter nearly in the ratio of 12 to 7. The interracial correlation of the mean numbers of stamens and pistils is very
much greater than the mean intraracial correlation between stamens and pistils,
being to the latter nearly in the ratio of 12 to 7. II.
Variation and Correlation in Lesser Celandine II. Variation and Correlation in Lesser Celandine 153 These conclusions show us how extremely cautious we must be in extending
conclusions deduced by intraracial calculations to interracial relations. Intra-
racial and interracial variabilities and correlatious may be of a totally diiFerent
order. By the usual theory of regression we have the following formulae to find
the probable number of stamens in a race given the mean number of pistils, and
the probable number of pistils given the mean number of stamens: Probable mean number of stamens in a) = -9939 x (observed mean number of
local race
j
pistils) + 7*185. Probable mean number of pistils in a) = "9106 x (observed mean number of
local race
J
stamens) — 4'697. Probable mean number of pistils in a) = "9106 x (observed mean number of
local race
J
stamens) — 4'697. Probable mean number of pistils in a) = "9106 x (observed mean number of
local race
J
stamens) — 4'697. Comparing with actual values we have : Comparing with actual values we have : Bordighera
St Peter Port
Surrey
Dorset
Early Belgian
Late Belgian
Trogen
Gais
Stamens
Obterved
26-7
280
32-9
35-6
26-7
17-9
20-4
238
from Pistils
Calculated
24-9
31-0
317
353
245
193
204
252
Pistils from
Observed
17-7
239
245
28-3
174
121
133
18-1
Stamens
Calculated
196
208
253
277
19-6
116
138
170 The maximum error in the number of stamens is 30 and the mean error 13;
for the number of pistils it is 3"1 and the mean error again 1#3. Thus our
formulae will give the number of stamens from pistils, or pistils from stamens
in any race with an average error of about one pistil or stamen, and a maximum
of about three. They are thus close enough for most practical purposes. The
diagram gives the lines to read stamens from pistils and pistils from stamens. (7) There are a number of other points which are suggested by our table,
but it seems better to defer their discussion until more ample data are forth-
coming. There is hardly any district from which at present a series of 500—600
celandine heads would not be of value. But these series ought to be gathered on
a uniform plan, Le. gathered as buds, each bud being wrapped in a separate soft
paper cover. II. Guernsey. I. Bordighera. Sepals. "5
(X,
6
S
9
io
11
Totals
_
2
—
2
S
1
3
521
46
10
1
682
k
_
6
19
14
—
39
5
_
—
1
—
1
Totals
1
9
542
60
11
1
624 I. Bordighera. Sepals. "5
(X,
6
S
9
io
11
Totals
_
2
—
2
S
1
3
521
46
10
1
682
k
_
6
19
14
—
39
5
_
—
1
—
1
Totals
1
9
542
60
11
1
624 III. Surrey. Sepals. CO
&
0)
PH
6
7
8
9
10
11
Totals
S
2
20
429
38
5
1
496
4
—
4
—
1
5
Totals
2
20
429
42
6
2
500
IV. Dorset. Sepals. Is
2of
7
8
9
10
11
IS
IS
Totals
S
4
279
115
59
31
3
—
491
4
2
4
1
1
1
13
5
_
—
—
—
1
1
Totals
4
283
117
63
32
4
2
606 III. Surrey. Sepals. CO
&
0)
PH
6
7
8
9
10
11
Totals
S
2
20
429
38
5
1
496
4
—
4
—
1
5
Totals
2
20
429
42
6
2
500 A. SEPALS AND PETALS. I. Bordighera. Sepals. "5
(X,
6
S
9
io
11
Totals
_
2
—
2
S
1
3
521
46
10
1
682
k
_
6
19
14
—
39
5
_
—
1
—
1
Totals
1
9
542
60
11
1
624
II. Guernsey. Sepals. als. CD
OH
7
8
9
10
11
Totals
S
6
450
49
7
3
616
k
1
2
1
—
1
6
Totals
7
452
50
7
4
620 II.
Variation and Correlation in Lesser Celandine Further since the labour of counting is very great, those who will
count as well as gather are the more valuable helpers. In counting great care
must be taken to separate the individual pistils, and all heads with less than
three sepals ought to be carefully examined with a microscope or powerful lens. If possible two series should be taken from each locality, one early and one late in
the season, but the flowers should not be taken off the same plants, but off plants,
having substantially the same environment. A record of date, environment and
ve s ty o Ju y , 0 5 (7) There are a number of other points which are suggested by our table,
but it seems better to defer their discussion until more ample data are forth-
coming. There is hardly any district from which at present a series of 500—600
celandine heads would not be of value. But these series ought to be gathered on
a uniform plan, Le. gathered as buds, each bud being wrapped in a separate soft
paper cover. Further since the labour of counting is very great, those who will
count as well as gather are the more valuable helpers. In counting great care
must be taken to separate the individual pistils, and all heads with less than
three sepals ought to be carefully examined with a microscope or powerful lens. If possible two series should be taken from each locality, one early and one late in
the season, but the flowers should not be taken off the same plants, but off plants,
having substantially the same environment. A record of date, environment and
y
y 30 30 Biometriia n Biometriia n Cooperative Investigations on Plants Cooperative Investigations on Plants 154 locality should accompany each aeries. The most important localities at present
seem to be, Mid-France, South and Mid-Italy, Mid and North England, Switzer-
land and South Germany in as many districts as possible, and any northern
European stations, where workers may be available. It would be of great value
to have one species of plant thoroughly worked out from the biometric standpoint
over a wide area. This can only be done by cooperative labour. III. Surrey. IV. Dorset. Sepals. Is
2of
7
8
9
10
11
IS
IS
Totals
S
4
279
115
59
31
3
—
491
4
2
4
1
1
1
13
5
_
—
—
—
1
1
Totals
4
283
117
63
32
4
2
606 II. Variation and Correlation in Lesser Celandine II. Variation and Correlation in Lesser Celandine 155 CO
&
5
O
O
"H a
O
n»
Totals
%
ss
ss
s§
i8
Sf
ss
IS
sr
0 0
04 04 i3i f"'
CO CO
1
-
I
1
CO
S3
S M
rH CO
CD
§
8 -
r- -H
|co
1«
| co m
1 ^ 1
I - 1
624
i - H
I—1
1
1
-
t—(
• *
• >
00
©3
CO
I - H
CO
CO
CO
i§
s
•>3
>
>
CO
•<*
I-H
<M
CO
a,
00
I - H
Totals
s
|
CO
Totals
33
%
* i
C>
*^
^
es
^H
0 0
'•o
*-(
>3
*^
• < » •
2"°
S
r—1
CO
co
CO
CO
©
CO
OJ
(N
TO i—
C»
c? 1
§ ^
f
CO
CM
S 1
CO
1
I - H
520
i - H
i
m
OC
CD
CD
O
OD
m
CM
SS
H
ro
CO
CO
3
CD
c
-11
r—<
CO
cq
1
1
Totals
•spdag
Total
"T
^
2$. -5-
•^v
c:
%
I I iT
•H" ° °
51
li
*? S? $
ai
-*
'-'
ca
•t
0
«
CO
O
X
CO
8
CO
- I f
41 | 33
s
CO
So
55
0;
94
i - H
09
r-H
r-H
CD
1
I - H Totals
33
%
* i
C>
*^
^
es
^H
0 0
'•o
*-(
>3
*^
• < » •
2"°
S
r—1
CO
co
CO
CO
©
CO
OJ
(N
TO i—
C»
c? 1
§ ^
f
CO
CM
S 1
CO
1
I - H
520
i - H
i
m
OC
CD
CD
O
OD
m
CM
SS
H
ro
CO
CO
3
CD
c
-11
r—<
CO
cq
1
1
Totals
Total
"T
^
2$. -5-
•^v
c:
%
I I iT
•H" ° °
51
li
*? S? III. Surrey. $
ai
-*
'-'
ca
•t
0
|
«
CO rH
O
rH
X
CO
8
CO
- I f
41 | 33
1
s
CO
So
1"*
55
0;
94
i - H
09
r-H
r-H
CD
1
I - H
-
500
r
f " i
0
! d
^ 1
i - H
i - H
CO
CO
8
8
(M
? 8
3
'^
en
CO
8
i - H
«
I - H
CD
- Totals
%
ss
ss
s§
i8
Sf
ss
IS
sr
0 0
04 04 i3i f"'
CO CO
1
-
I
1
CO
S3
S M
rH CO
CD
§
8 -
r- -H
|co
1«
| co m
1 ^ 1
I - 1
624
i - H
I—1
1
1
-
t—(
• *
• >
00
©3
CO
I - H
CO
CO
CO
i§
s
•>3
>
>
CO
•<*
I-H
<M
CO
a,
00
I - H
Totals
s
•spdag |
CO VI Stamens. *5
34
2
36
S6
26
1
26
57
29
29
SS
37
3
40
39
22
22
4
2
2
LS AND PIST
Bordxghera. Pistils. 22
24
1
26
SS
14
1
15
24
6
6
25
7
7
«6
1
1
Ghternsey. Pistils. 50
13
13
31
11
11
SS
19
19
SS
16
16
54
5
5 3
4
5
Totals
IS
1
1
141616171819SO
2
2
SI
1
28
1
i
1 1
SS
6
6
24
6
6
25
7
7
26
8
8
27
8
8
28
14
14
29
17
17
SO
23
23
SI
20
20
a? 35
35
S3
47
1
48
S4
36
36
5
34
2
36
S6
26
1
26
57
29
29
SS
37
3
40
39
22
22
40
29
29
C. SEPALS AND PISTIL
IX. Bordxghera. Pistils. 2
S4
6
Totals
6
1
1
7
—
—
8
—
—
9
—
—
10
1
1
11
3
3
9
9
IS
24
24
63
5
68
15
71
4
76
i6
76
7
82
17
82
3
86
18
1
71
7
79
19
1
60
6
56
20
31
2
33
*2
31
3
34
22
24
1
26
SS
14
1
15
24
6
6
25
7
7
«6
1
1
8
6
6
X. Ghternsey. Pistils. 5
4
Totals
18
6
5
13
4
4
U
10
10
i5
17
17
16
16
1
16
17
28
28
iS
32
32
19
28
28
*0
28
28
21
33
1
34
22
35
36
«5
30
30
24
30
1
31
85
40
40
*6
23
1
24
27
28
28
28
20
20
«9
16
16
50
13
13
31
11
11
SS
19
19
SS
16
16
54
5
5
y
y ,
p
j
g II. Variation and Correlation in Lesser Celandine II. Variation and Correlation in Lesser Celandine 157 3
Totals
$
3-
$
- « * •
ss
85
«0
g
sr
$
= 0
f~
*"H
to
*-H. "5
.«0
s
*-H. *H
*"H. o o
O5
•
*
*
- 1
- 1
*
! |
CO - 1
1 1
•>f r-H
eo
m •->
CD
rH
i-H
o
CO
|
00 -H
2
<M i—I
i-H
t ~
£ 1
£ 1
m
i—«co
l - H
3 1
it
3
3 1
I-H i-H
«
S 1
2
* 1
« 1
- 1
600
i - H
i - H
i
1-H
1
•
*
•o
to
• *
to
•
*
CO
rH
a>
i - H
CO
i—i
1-H
<N
i—I
s
CO
i-H
CO
I - H
s
CD
O
f-H
o
oq
Totals
Dorset. rtils. Totals
91S9
99-t9
o
13
c>
-<r
$
to
I? ss
8
*H. 0 0
c -
*-H
to
*H
«O
*-H
• 3
* f
*M
" 1
*-(
O
>-^
OJ
491
1
1
1 -
1
- 1
1 1
1 1
1
-
- 1
• *
f - H
r - H
I
< 1
1
1
(N i-H
|
- * r-H
|
00
I
t~ i-H
|
1
00 rH
|
03
0 0
I
1
« - . ,
s
2
1
1
1
lO rH i
r-H
<
1
2
3
1
1
1
S^ i
Srt i
Srf i
S~ i
2
5
t -
oo
o
o>
a*
r-H
1
f - H
* 1
1
1
1
605
r-H
1
i - H
1
l - H
1
i - H
1
1
I - H
i - H
• *
CO
•o
0 0
0 0
Ti"
a
a
CO
•*e
i-H
r-H
CO
f -
r-H
CD
3
00
«
CD
CO
t~
CO
c. §
g
00
r-H
•o
l-H
t-
l—1
t ~
00
o
a*
l - H
M
i—I
r-H
1
Totals
02
H
CO
Bordigh
Stamens. |
CO 2 a
U
P4
XI
Totals
%
- * » •
4-
ss
is
If
§5
S3
sr
8
OS
* 1
00
>H
fS
rH,
to
HaMQHri
>o
1
1
-
-
1
- 1
COr-rH
COrH-
COrH
CO
-
CO • *
O
I> r-H
CO rH rH
CO
co cnoo
t - CO «
CO
HQ0«
rH
-2~
!-
1
t-
«
«
rH
* -
I
1
624
1
i
1
r -
r H
" *
O
oo
00
m
rH
•o
CD
00
t -
s
8
CO
r-H
CO
CO
r-t
a
CD
rH
r H
Totals
at Carleton University on July 1, 2015
http://biomet.oxfordjournals.org/
Downloaded from Totals
%
- * » •
4-
ss
is
If
§5
S3
sr
8
OS
* 1
00
>H
fS
rH,
to
HaMQHri
>o
1
1
-
-
1
- 1
COr-rH
COrH-
COrH
CO
-
CO • *
O
I> r-H
CO rH rH
CO
co cnoo
t - CO «
CO
HQ0«
rH
-2~
!-
1
t-
«
«
rH
* -
I
1
624
1
i
1
r -
r H
" *
O
oo
00
m
rH
•o
CD
00
t -
s
8
CO
r-H
CO
CO
r-t
a
CD
rH
r H
Totals
at Carleton University on July 1, 2015
http://biomet.oxfordjournals.org/
Downloaded from Stamens. XIV. Guern $
CD
7
9
10
11
Totals
1
1
—
—
—
—
2
2
3
3
1
1
10
11
16
16
28
1
29
38
\
40
4
28
1
29
36
1
38
36
61
2
63
38
5
46
39
4
43
23
2
25
SO
28
3
31
23
8
31
1
18
4
1
24
55
1
19
3
1
24
17
4
1
22
16
2
1
18
4
3
2
1
XV. Surrey. Stamens. at
6
7
8
9
10
11
Totals
18
1
1
19
1
1
SO
E
—
—
—
SS
6
6
SS
1
10
11
«4
2
2
S5
1
13
14
S6
1
28
29
rr
19
1
20
3
29
2
i
36
S9
2
23
3
29
SO
1
26
1
27
31
4
36
3
42
SS
2
26
2
1
31
S3
28
1
1
30
5.*
\
38
2
41
55
29
4
33
SG
2
22
24
57
2
17
4
23
S8
19
3
23
59
16
4
20
11
1
I
13
6
3
8
4S
CO 09
11
XVI. Dorset. Stamens. 7
8
9
10
11
IS
IS
Totals
ts
1
1
H 1516171819SO
2
2
SI
1
1
SS
1
1
S3
a
6
*4
6
6
£5
6
2
7
6
1
1
8
£7
6
1
1
8
S8
12
1
1
14
«9
1
11
4
1
SO
17
6
1
17 23
SI
13
5
1
1
20
SS
2
23
7
2
1
36
S3
1
36
6
5
2
48
54
22
6
7
2
36
55
20
7
6
3
36
36
14
6
4
2
26
57
17
8
3
1
29
38
19
14
3
3
1
40
39
10
9
3
22
40
13
10
4
2
29
41
8
3
4
2
1
18
4*
4
7
3
2
16
45
8
4
4
2
18
44|4
1
2
2
2
1
8
2
3
1
6
at Carleton University on July 1, 2015
http://biomet.oxfordjournals.org/
Downloaded from II. Variation and Correlation in Lesser Celandine II. Variation and Correlation in Lesser Celandine 159 ag
h-1
Q
Bordighe
Pistils. Stamens. Totals
*»,
to
g
Cd
sr
$
00
l-H
CO
•5
»*H
o
00
CO
•*!Dr-
o
1
1
1
1
-
1
1
~,-<rH
|
1
- 1 1
1
-* 1
1 -
1
- 1
1 1
1 1
1
1
^H f-t
co
1
1
CD rH
CO
CO r-H rH I
l-H
1
1
rHOJKjj CO
1
C O g O , | r H
1 g-^r-
oa eo co rH
1-1 £ *""
rHg««
1
1
CO i—1
m
CO
1
1
l « ~ 1 1
1 1
1 1 1 1
1
1 1
1 1 1 1
1 - 1
1
CM
to
i—i
n
at
i—i
1
-
at
o
CD
2
S
s
g
a>
s
ao
s
CD
eo
1
1
otals
Totals
00
ss
5? sr
S3
CO
1? • 5
! «
l«
1
1
| | Wr-rH
1
1-
- 1
1 1 1
«
|
rH CO
1
1
1
| ^ . r H
| rH
co« | |
|cOr-r- |
^ r H
|
|
S * 1 1
1 rH
rH eo at
1
1
IS0"" 1
S w 1 1
rH CO <O
rH oo at
at
1
1
gCT 1 1
H r i ^
1
I8-""
15^
IS
1
1
CO
I •-<
12 1 1
1" 1
1-= 1 1 1
620
r—t
1
1
1
1
at
1
1
«
1
at
CO
o
»
iO
rH
cb
eo
l-H
CO
g
«
eo
ct
CD
rH
t-
i—1
o
l - l ag
h-1
Q
Bordighe
Pistils. Stamens. Totals
*»,
to
g
Cd
sr
$
00
l-H
CO
•5
»*H
o
00
CO
•*!Dr-
o
1
1
1
1
-
1
1
~,-<rH
|
1
- 1 1
1
-* 1
1 -
1
- 1
1 1
1 1
1
1
^H f-t
co
1
1
CD rH
CO
CO r-H rH I
l-H
1
1
rHOJKjj CO
1
C O g O , | r H
1 g-^r-
oa eo co rH
1-1 £ *""
rHg««
1
1
CO i—1
m
CO
1
1
l « ~ 1 1
1 1
1 1 1 1
1
1 1
1 1 1 1
1 - 1
1
CM
to
i—i
n
at
i—i
1
-
at
o
CD
2
S
s
g
a>
s
ao
s
CD
eo
1
1
Totals Tota
00
ss
5? sr
S3
CO
1? • 5
! «
l«
1
1
| | Wr-rH
1
1-
- 1
1 1 1
«
|
rH CO
1
1
1
| ^ . r H
| rH
co« | |
|cOr-r- |
^ r H
|
|
S * 1 1
1 rH
rH eo at
1
1
IS0"" 1
S w 1 1
rH CO <O
rH oo at
at
1
1
gCT 1 1
H r i ^
1
I8-""
15^
IS
1
1
CO
I •-<
12 1 1
1" 1
1-= 1 1 1
t1* 00 Oi Ci >H
620
r—t
1
1
1
1
at
1
1
«
1
at
CO
o
»
iO
rH
cb
eo
l-H
CO
g
«
eo
ct
CD
rH
t-
i—1
o
l - l
Totals ag
h-1
Q
Bordighe
Pistils. Cooperative Investigations on Plants 160 Totals
g
91-89
99^9
? •3
•
a
%
ss
18
2"
£
Sg
OS
CO
^ *
o
CT>
^
CC t ^ CO Cfl ^
0*1
CD i-H Cp CO
OT f-^
1 1 1 1 1 1 rH
1 1 1 1 1 1
1
1 1 1 1 rH | |
1 1 1 1 1 1
1
1
1
1
1
1
1
1
1 rH | |
1 1 1 1 1
- | 1 1 |
1 1 1 1 1
1 1 1 I 1
rH | | | |
1 rH | | | | |
| H | H « | 1
in mi i i
1 rHrH | | | |
| 09 rH | rH | H
| CO rH 11" | | |
| <N Ot Ot r-l rH |
| O, -H rH | | |
1 • * • *
1 r - 1 I
I B M I * 1 1
| CO CO 1 M I I
1 cot- •>* I | I
lonn* I |
| t-HlO | | |
1 O-*rH« , ,
|
C O • * • > * 1 1 |
rH CO CD CO | | |
| • * 05 « 1O i-H |
^S"""" 1 1 1
| « O « H | |
1 gco-oco | ,
1 co^eo | ~ |
| t-I-iO | | |
rH 1O CO • * « | |
l-H CO C— rH rH i-H ,
l-H
1
i 8 N I " i i i
i SWrH i i i
12^1^1 i
i 2 " ^
1 CO | rH
| >
| rt
1 - * 1 rH
1 at |
1 ^ 1
1 at 1
1 1 «
1 rH |
1 1 1
1 rH |
1 1
I |
1 1
1 1
1 1
1 1
1 1
1 1
1 1
1 1
1 1
t» oo 01 O »H. ag
h-1
Q
Bordighe
Pistils. • } 03
606
1
r-l
1
i - H
1
r H
1
1
l - H
r-l
co
at
>a
CO
co
•d>
ct>
a>
CD
T-*
I - H
CO
5
2
%
3
CD
I-H
3
3
3
3
00
l-H
r-
r-
co
•o
o*
I - H
CH
i-*
I
l - H
[Totals 00
Totals
3-
$
3-
3-
%
Sg
ig
S5
$
3
3T
8
SI
$
OS
i-i
00
» 1
5? at^^at
- 1
1
01
1 1
1 1
1
m <->
| rH CO rH
1
1
1
CM rH
CO rH
1
« C 0 | r H
• *
O rH
l-H
l-H M
l-H
1
HMHH
l-H
at
I - H
at • * rH
rH
CC^rH
a™ i
CO CO «
at
- & w 1
l-H f - CO
at
CO
1 rH CO
1 CO
rnrngCO |
rt8M
1 1
1 M S r t
cog
eo t~ «
l-H
1 12
1
1
OS i-H
o
CO
1
~« 1 1 1
1
9 1
|
|
• *
—S3
500
-
i - H
cq
i
i-H
1
o
CO
CO
i - H
o
CO
rH
a>
o
r-l
CO
rH
at
l-H
CO
CO
rH
CO
l - H
ct
a
s
Ol
at
•o
o
l-H
o
CO
^ . 91
Totals 00
Totals
3-
$
3-
3-
%
Sg
ig
S5
$
3
3T
8
SI
$
OS
i-i
00
» 1
5? ag
h-1
Q
Bordighe
Pistils. at^^at
- 1
1
01
1 1
1 1
1
m <->
| rH CO rH
1
1
1
CM rH
CO rH
1
« C 0 | r H
• *
O rH
l-H
l-H M
l-H
1
HMHH
l-H
at
I - H
at • * rH
rH
CC^rH
a™ i
CO CO «
at
- & w 1
l-H f - CO
at
CO
1 rH CO
1 CO
rnrngCO |
rt8M
1 1
1 M S r t
cog
eo t~ «
l-H
1 12
1
1
OS i-H
o
CO
1
~« 1 1 1
1
9 1
|
|
• *
—S3
500
-
i - H
cq
i
i-H
1
o
CO
CO
i - H
o
CO
rH
a>
o
r-l
CO
rH
at
l-H
CO
CO
rH
CO
l - H
ct
a
s
Ol
at
•o
o
l-H
o
CO
^ . 91
Totals
I -
* -a
OH
Tot
g
91-89
99^9
? ag
h-1
Q
Bordighe
Pistils. •3
•
a
%
ss
18
2"
£
Sg
OS
CO
^ *
o
CT>
CC t CO Cfl
0 1
CD i-H Cp CO
OT f-^
1 1 1 1 1 1 rH
1 1 1 1 1 1
1
1 1 1 1 rH | |
1 1 1 1 1 1
1
1
1
1
1
1
1
1
1 rH | |
1 1 1 1 1
- | 1 1 |
1 1 1 1 1
1 1 1 I 1
rH | | | |
1 rH | | | | |
| H | H « | 1
in mi i i
1 rHrH | | | |
| 09 rH | rH | H
| CO rH 11" | | |
| <N Ot Ot r-l rH |
| O, -H rH | | |
1 • * • *
1 r - 1 I
I B M I * 1 1
| CO CO 1 M I I
1 cot- •>* I | I
lonn* I |
| t-HlO | | |
1 O-*rH« , ,
|
C O • * • > * 1 1 |
rH CO CD CO | | |
| • * 05 « 1O i-H |
^S"""" 1 1 1
| « O « H | |
1 gco-oco | ,
1 co^eo | ~ |
| t-I-iO | | |
rH 1O CO • * « | |
l-H CO C— rH rH i-H ,
l-H
1
i 8 N I " i i i
i SWrH i i i
12^1^1 i
i 2 " ^
1 CO | rH
| >
| rt
1 - * 1 rH
1 at |
1 ^ 1
1 at 1
1 1 «
1 rH |
1 1 1
1 rH |
1 1
I |
1 1
1 1
1 1
1 1
1 1
1 1
1 1
1 1
1 1
t» oo 01 O »H. ag
h-1
Q
Bordighe
Pistils. • } 03
606
1
r-l
1
i - H
1
r H
1
1
l - H
r-l
co
at
>a
CO
co
•d
ct>
a>
CD
T-*
I - H
CO
5
2
%
3
CD
I-H
3
3
3
3
00
l-H
r-
r-
co
•o
o
I - H
CH
i-*
I
l - H
[Totals 1 cot- •>* I | I
T-* | « O « H | |
3 £
1 gco-oco | , Sg
1 co^eo | ~ | | t-I-iO | | |
3 rH 1O CO • * « | | i 8 N I " i i i
3 II. Variation ami Gorrdatioii in Lessen- Celandine 161 •diglier
Is. 5 S
°3 2
PL! Totals
^
is
83
SS-
91
sr
i? ti
Si
^•i
= 0
8~
CO
" I
>c
* s
*-<
©i*-i
—i
o
>-(
0 0
t^
CO
1
1 1 1
1
1—1
I
1 1
1 1 1
1
1
1
1
r—1
1
1 1 - 1
I—t
-
I - H
l - H
t—t
I - H
^H
r—>
l - H
I - H
I—t
1—1
r-H
! 1
1
1
1 ! 1
I 1 1
1 1
" 1
1
H >-H
f» r-H
1
i - H
0 ) CM
l - H
1 1
H | «
1
1
1 1
1
1
1 1 1
I
I - H
Cft ri
(N | ® iO C5 •<* CM
1
I
"OWOHiOO)nO^OH
I-H
ri iHtHiHnt-BrfOOl^CD-tOl
1
HMHrecoM-Hrtaan
i—4
I-M
I-H
,-H
1
•
<
*
r-i
1
l - H
i
1 1
1
1
1
1
1
1
1
1
1 1 1
1 1 1 1
1 1 1 1
- 1
1 I
~
1
1
1
1
1
1
1 1
1
1 1
1
1
1
1
1
1
1 1
1
1 1
1
1 1 1
I - H
1
1 i
i
i—i i—<
1
1
1
I
-
t
55 E;55Sa§{llsr8§i;S?§§ BOSS'S SfeSS ^ ^ ^ ^ 5 1 . ag
h-1
Q
Bordighe
Pistils. ^
624
i - H
1
1
n
(X
i—i
1
1
i—1
i—(
CM
O
H
l -
CD
O
3
S
1 ~
cc
01
X
§
•*
OS
n
1
1
1
f t
Totals
at Carleton University on July 1, 2015
http://biomet.oxfordjournals.org/
Downloaded from •diglier
Is.
5 S
°3 2
PL! 31 Biometriia n Biometriia n Cooperative Investigations on Plants 162 Total
vs
•*>
oc
-«]•
!~
• * » •
to
$
- « ) •
-<r
91
- < ? •
»-i
- • » •
- > ! •
3
1? ^H
»5
O
Si
$
5? $
Si
$
* - i
0 0
CO
i-1
-«J-
i-H
•2
i-i
» )
r-*
1
1
1
1
1
1
1
I
1
I
1
1
1
1
1
1
1
1
1
1
1 1
1
1
1
1
1
1
1
1
1
1
1
1 1
1
1
1
1
1
1—I
co
04
1
1 1
-
1 -
(W
rH
-
O» CO
1
1
1
t - H
1-H
rH | ^
0 9
r H
- 1
at
1-1«
1-1 rt «
~H
09
- H CO
1—1
i - H
1
1 1
1—1
1 1
1
09 |
r -
i - H
r H
r H
1—' ^ H
I - * 1—< (—1 I-H
1
1
-
-
1
1
^ 1 1 1 1
- 1
(71 O4
r-H
I-*
l - H
1
I - H
1-H
•fll rt 1-H rH
I-H 1-H 09 1-H « -H
•— ~H CO 09
l-H
H
1— l-H i-H Tf
I - H
I - H
1—t
! •diglier
Is.
5 S
°3 2
PL! < *
n«HH^f(O«rHHH«H
t3
.-H CO O4 CO
1
i 04
r-H CO
to co co —1 0 ri n
« « ^H
rH-H
09 i-H rH 09 M rH
|
1 1 1 -
1 1
i-H
r-*
O9 rH 09 O9 rH
nnnnnftoiKrH | r-«
Tf O) -4< 09 CO 09
0 9
Ot O9 09 09
| «ntDiHRn««nn |
^
| i-Hnooro^fi-irti-'-*
1
09 CO i-H •<» CO
I-H
CO 04
1—t
i-c at
1 «
H O ) « H
l-H
l—l
l-H
1—1
1
1
I - H
09 rH
r -
1 1 1 1
1
1 1
i - H
1
1 1
1-
1
1
1 1 1 -
1
1
i
1
1
1
1
1
1
1
I - H
1
1
1
1
1 1 1
1
1
520
r -
1
I
1
f
O9
1
T *
1
O9
1
O9
CO
<o
1 0
10
I-H
a>
l-H
CO
i-H
CO
i-H
3
0 9
O
rH
CO
C? 3
S
09
CO
3
CO
t -
0
I-H
•a
Totals II. Variation and Correlation in Lesser Celandine 63 2
&
x
X
tils. Pis
Totals
3-
w
. •diglier
Is.
5 S
°3 2
PL! 5f
§
CO
00-I
• H
1-1
S,
o
I-H **
1
i
i
i
i
I
i
i
i
i
1
1
I
—
1
-
1
1
- 1
I i ~ -
I
i
- 1
1 1 1
I
1
-
1
1
i - H
1 1 1
1
1
1 1
I-H
r-H
i-H
1 1
•-
1
1
1 1
l-H
i-H
l - H
1
1
rH f r -
I-H f^
r-^
l-H
rH
|
«rHrH
l-H r^
-
| -
r H _
-
CO
CM rH
CO
l - H
Q* <-4 i-l
l-H 04 i-H i—1 i-H
OS
CO
rH rH „ r- C, r- r-
CO
TJ" rH 0, rH r i « W rH r-
rH CO « O CO r- « CO « « « rH
I-HOT]
1 » -<f (» M O H rH
rH OJ l-H rH i-H Tf n
•-I CO <»
- I I I
i—i
rH
l - H
1
-
1
l - H
rH r~i
-
, , r- CO T, -H r- « r- rH
«rH««T,.«^COCO«rHrH | rH
rH « O CO « «, rH
«
CO .H COCO
««rH
<OCOT,CCO | r . H H |
00 CO CO W CO »O CO
rH i-H O4I ^ * rH
CO
1
I—I
C0 « CO rH rH
-
- 1
coeo-HCOMeowwrHrH^,,-
r™< d
1 rH I r^ *O 04 W CO 04 O4J
^H 1 1 O,} CO 04 rH 04 rH Or! •diglier
Is.
5 S
°3 2
PL! I rH
|
I
-
-
| e o W r H
1 «
I I I - I
1 1
1 1
rH rH
rH rH
- 1
rHrH | |
| r H , -
1
1
1 -
r H
1
1
1
1
1 1
r H
r-H
1
1 1
1
1
1
1
1
1
1 1 1 1
-
1
1
1
1
1
r H
-
- -
-
1 1
r H
1
-
l - H
1 1
1- 1
1
1
1
1
1
1
1 1 1 1
1
1
1
1
1
1
1
1
i
1 1
1
1
1
1
1
1
600
-
O
r H
•
*
*
*
CO
T »
OS
T f
l - H
•o
2
o»
•o
CO
I-H
: :
Oi
r*
CO
s
eo
rt
%
3
OS
s
04
(?)
•a
rH
o
<=
CO
Totals
at Carleton University on July 1, 2015
http://biomet.oxfordjournals.org/
Downloaded from 21—2 Cooperative Investigations on Plants 164 i i i i i
i i i i i
i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i I i i i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i i i i
i i i i i i i i i i i - i i
i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i
i i i i i i
i i i i i i i i
i i i i i i i i i - i i i i i i i i
i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i
I I I I I I I I I I I I I I I I ! •diglier
Is.
5 S
°3 2
PL! I
I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I -
1 I I
I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I
I I I I I I I I - I I I I I I I I I
I I I I I I I I I I I I I I
i i i i i i i i i i i i i i i
i i i i i i i i i i i i i - •
i i i
I I I I I I I I I I I I I
i i i i i i i i i i i i i i i
i i i i i i i i - i i i i i i
I I I I I I I I I I I I I I
i i i i i i i i i i i i i i i
i i i i i i i i i i i i i I - I
I -
i i i i i i i i i i i i i i i
i i i i i i i i i i i i
I I I I I I I I I I I I I I
I I I i I I I I I I I I I I
i i i i i i i i i i i i i i i
i i I H I i i i i i
- I - I I I I I I I I I I I
I I I I I I I I I I I I I I I
I I I I I I I I I
- I
I I I I I I I I I I I I
oo
00
I I I I I I I I I I I I I I I
I I I I I I I I I I I <N ~
I I I
I I I I
I I I
I I i
I I I
I I II I
_l I l_
I I I
I I I I l - l
I -
| M | - « I -
| I - I
I I
I I
I I I I J _ l ^ I _ J _ _ [ _
i i i Y i i I i i
I I I I - I I
I I
I I -
I I - I I I I I 1 I I Ii i
I I I I I I I I I I I I I I I
I 1 - 1 I I I I I | H . •diglier
Is.
5 S
°3 2
PL! I I
I I I I I I I I I I I
Ji-
lt"
I I I I I I I I I I I I I I I
I I I I I - » I - 1 " I I < N « - I I I I I - I I I I I I I I I Ii i
i i
I I I I I I I I I I I I I I I
I I I - I I I -
-
r
e
- re I
I I I - I I I I I I I I I I I
I I I I I I I I I I I I I I I
I | |
I I I 1 - 1 I I I |
H
| I I I I I I I I I
CO
A. 35
I I I I I I I I I I I I I I I
I I | H I
• - H i
I - I
I I I
I I I I I I I I
I I I I I I I I I I 1 I I I I
— I . •diglier
Is.
5 S
°3 2
PL! I
— I | | I | — I I
I I
I I
I I I I I I I I I I I I I—
— • * » I I
I I I
1 J_L
i i i
I - I I I I I I I I I I
I I
I I I I I I I I I I
i i
i i
i \ i i
i i i i
i i i
i i i
I
I
|
I
I
I
I
I
I
I
I
| 0 0 CO C74 I O« -4< .—t •—r .—i ,—• 1 I
I
|
I
I
|
I
I •—i I
I
I
I
I
I
I I 1
I
l
l
l
l
l
l
— — —
l l r e — w r e M —
re-tf»io»—
i
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
I I I I I I I I I I I I l _ J _ j * > « " » _ ] _
— I I I I I I I I I I I I I I I I c o
I— O T » ( D W * H I—«
I re I — I I I I I I I I I I I I I I
— I
I I I I I I I I I I I I I I I I I
I I
I I
I I
I I I I I I I I I I I I — I — I
l«MOM5)
— If | I — | — —
I
I
I
I I
I
l
l
l
l
l
l
l
—
M
i
l
—
— l O f n - H M O - J O l M M M
I | | H I I I I I I I I I I I I I I I I I I
I I I I I I | I I I m
| H i n
I
I I I I I I I I I I I I I I I
I I I I I
JL
O5
00
I
l
l
l
l
l
l
l
l
l
l
—
« — re —
— i i —
l l l l l l l l l l « l l l l
© M
- * - * w - l —
I
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
1.1 I I
I I I I
I I I
777"
I
'
l
l
l
l
—
I — — — - f i r e — —
I
I
W
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I I | M W n r t H M |rH | H I— — — I— I I
I I I I I I I I I I I I I I I I I I
I
l
l
l
l
l
l
l
l
l
l
l
l
—
I — I — I — — I I <» I I I I I I I I I I I I I I I I I I I I I I I I I
I I I I I I I I I I I
I I I I —
w —
I I I I I I I I I I I ! •diglier
Is.
5 S
°3 2
PL! I I I I I I I I I I I I I
CO
o
04
I f i
•—• •—• >—• • •—• •—•
[
i
i
i
i
l
i
i
l
f
i
i
i
i
r
i
i
t
f
r
I I I I I I I I I I
I i i i i i i i I I -
i I I i i i i i i i i -
I i I i I i i I i i i i i I i i I i I i
i I i I i
I I i. •diglier
Is.
5 S
°3 2
PL! i I i I i i -
i I i I
I I I l I I I I I I
I l I I
I I I
I — I
I I I I I I I I I I I I I I I I I I I I I
I I I I I
I I I - I I I I I I I I I I I I I I I I I I I I I I I I I I I
l I I l
I I
I I I |
H
|
I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I I I I I
i i i
i i i - i i i i i i i i i i i i i i i i i i i i
O5
i i i i
I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I
i i i i i i i - i i i i i i i i l i i i I i i i i i i i i i i i i i i i i i i i i i I
i i i i i
at Carleton University on July 1, 2015
http://biomet.oxfordjournals.org/
Downloaded from i i i i i
i i i i i
i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i i I i i i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i i i i
i i i i i i i i i i i - i i
i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i i i i
i i i i i i i i i i i i i i
i i i i i i
i i i i i i i i
i i i i i i i i i - i i i i i i i i
i i i i i i i i i i i i i i
i i i i i i i i i i i i i i i
I I I I I I I I I I I I I I I I ! •diglier
Is.
5 S
°3 2
PL! I
I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I -
1 I I
I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I
I I I I I I I I - I I I I I I I I I
I I I I I I I I I I I I I I
i i i i i i i i i i i i i i i
i i i i i i i i i i i i i - •
i i i
I I I I I I I I I I I I I
i i i i i i i i i i i i i i i
i i i i i i i i - i i i i i i
I I I I I I I I I I I I I I
i i i i i i i i i i i i i i i
i i i i i i i i i i i i i I - I
I -
i i i i i i i i i i i i i i i
i i i i i i i i i i i i
I I I I I I I I I I I I I I
I I I i I I I I I I I I I I
i i i i i i i i i i i i i i i
i i I H I i i i i i
- I - I I I I I I I I I I I
I I I I I I I I I I I I I I I
I I I I I I I I I
- I
I I I I I I I I I I I I
oo
00
I I I I I I I I I I I I I I I
I I I I I I I I I I I <N ~
I I I
I I I I
I I I
I I i
I I I
I I II I
_l I l_
I I I
I I I I l - l
I -
| M | - « I -
| I - I
I I
I I
I I I I J _ l ^ I _ J _ _ [ _
i i i Y i i I i i
I I I I - I I
I I
I I -
I I - I I I I I 1 I I Ii i
I I I I I I I I I I I I I I I
I 1 - 1 I I I I I | H . •diglier
Is.
5 S
°3 2
PL! I I
I I I I I I I I I I I
Ji-
lt"
I I I I I I I I I I I I I I I
I I I I I - » I - 1 " I I < N « - I I I I I - I I I I I I I I I Ii i
i i
I I I I I I I I I I I I I I I
I I I - I I I -
-
r
e
- re I
I I I - I I I I I I I I I I I
I I I I I I I I I I I I I I I
I | |
I I I 1 - 1 I I I |
H
| I I I I I I I I I
CO
A. 35
I I I I I I I I I I I I I I I
I I | H I
• - H i
I - I
I I I
I I I I I I I I
I I I I I I I I I I 1 I I I I
— I . •diglier
Is.
5 S
°3 2
PL! I
— I | | I | — I I
I I
I I
I I I I I I I I I I I I I—
— • * » I I
I I I
1 J_L
i i i
I - I I I I I I I I I I
I I
I I I I I I I I I I
i i
i i
i \ i i
i i i i
i i i
i i i
I
I
|
I
I
I
I
I
I
I
I
| 0 0 CO C74 I O« -4< .—t •—r .—i ,—• 1 I
I
|
I
I
|
I
I •—i I
I
I
I
I
I
I I 1
I
l
l
l
l
l
l
— — —
l l r e — w r e M —
re-tf»io»—
i
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
I I I I I I I I I I I I l _ J _ j * > « " » _ ] _
— I I I I I I I I I I I I I I I I c o
I— O T » ( D W * H I—«
I re I — I I I I I I I I I I I I I I
— I
I I I I I I I I I I I I I I I I I
I I
I I
I I
I I I I I I I I I I I I — I — I
l«MOM5)
— If | I — | — —
I
I
I
I I
I
l
l
l
l
l
l
l
—
M
i
l
—
— l O f n - H M O - J O l M M M
I | | H I I I I I I I I I I I I I I I I I I
I I I I I I | I I I m
| H i n
I
I I I I I I I I I I I I I I I
I I I I I
JL
O5
00
I
l
l
l
l
l
l
l
l
l
l
—
« — re —
— i i —
l l l l l l l l l l « l l l l
© M
- * - * w - l —
I
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
l
1.1 I I
I I I I
I I I
777"
I
'
l
l
l
l
—
I — — — - f i r e — —
I
I
W
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I
I I | M W n r t H M |rH | H I— — — I— I I
I I I I I I I I I I I I I I I I I I
I
l
l
l
l
l
l
l
l
l
l
l
l
—
I — I — I — — I I <» I I I I I I I I I I I I I I I I I I I I I I I I I
I I I I I I I I I I I
I I I I —
w —
I I I I I I I I I I I ! •diglier
Is.
5 S
°3 2
PL! I I I I I I I I I I I I I
CO
o
04
I f i
•—• •—• >—• • •—• •—•
[
i
i
i
i
l
i
i
l
f
i
i
i
i
r
i
i
t
f
r
I I I I I I I I I I
I i i i i i i i I I -
i I I i i i i i i i i -
I i I i I i i I i i i i i I i i I i I i
i I i I i
I I i. i I i I i i -
i I i I
I I I l I I I I I I
I l I I
I I I
I — I
I I I I I I I I I I I I I I I I I I I I I
I I I I I
I I I - I I I I I I I I I I I I I I I I I I I I I I I I I I I
l I I l
I I
I I I |
H
|
I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I
I I I I I I I I I I I I I I I I I I I
i i i
i i i - i i i i i i i i i i i i i i i i i i i i
O5
i i i i
I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I
i i i i i i i - i i i i i i i i l i i i I i i i i i i i i i i i i i i i i i i i i i I
i i i i i
at Carleton University on July 1, 2015
http://biomet.oxfordjournals.org/
Downloaded from
|
https://openalex.org/W2902071825
|
https://ora.ox.ac.uk/objects/uuid:36a9ba18-737d-42bd-8d7e-c8a8d7ae96ba/download_file?safe_filename=Nationwide%2Bschool%2Bmalaria%2Bparasitaemia.pdf&file_format=application%2Fpdf&type_of_work=Journal+article
|
English
| null |
Nationwide school malaria parasitaemia survey in public primary schools, the United Republic of Tanzania
|
Malaria journal
| 2,018
|
cc-by
| 11,520
|
Abstract Background: A nationwide, school, malaria survey was implemented to assess the risk factors of malaria prevalence
and bed net use among primary school children in mainland Tanzania. This allowed the mapping of malaria preva-
lence at council level and assessment of malaria risk factors among school children. Methods: A cross-sectional, school, malaria parasitaemia survey was conducted in 25 regions, 166 councils and 357
schools in three phases: (1) August to September 2014; (2) May 2015; and, (3) October 2015. Children were tested for
malaria parasites using rapid diagnostic tests and were interviewed about household information, parents’ education,
bed net indicators as well as recent history of fever. Multilevel mixed effects logistic regression models were fitted to
estimate odds ratios of risk factors for malaria infection and for bed net use while adjusting for school effect. Results: In total, 49,113 children were interviewed and tested for malaria infection. The overall prevalence of malaria
was 21.6%, ranging from < 0.1 to 53% among regions and from 0 to 76.4% among councils. The malaria prevalence
was below 5% in 62 of the 166 councils and above 50% in 18 councils and between 5 and 50% in the other councils. The variation of malaria prevalence between schools was greatest in regions with a high mean prevalence, while the
variation was marked by a few outlying schools in regions with a low mean prevalence. Overall, 70% of the children
reported using mosquito nets, with the highest percentage observed among educated parents (80.7%), low land
areas (82.7%) and those living in urban areas (82.2%). Conclusions: The observed prevalence among school children showed marked variation at regional and sub-
regional levels across the country. Findings of this survey are useful for updating the malaria epidemiological profile
and for stratification of malaria transmission by region, council and age groups, which is essential for guiding resource
allocation, evaluation and prioritization of malaria interventions. Keywords: Malaria, School children, Malaria surveillance, Malaria prevalence, Mosquito net use, Tanzania *Correspondence: chackyfa@gmail.com; manuela.runge@swisstph.ch
†Frank Chacky and Manuela Runge contributed equally to this work
1 Ministry of Health, Community Development, Gender, Elderly,
and Children, Dodoma, Tanzania
3 Swiss Tropical and Public Health Institute, Basel, Switzerland
Full list of author information is available at the end of the article © The Author(s) 2018. Chacky et al. Malar J (2018) 17:452
https://doi.org/10.1186/s12936-018-2601-1 Chacky et al. Malar J (2018) 17:452
https://doi.org/10.1186/s12936-018-2601-1 Malaria Journal Open Access Nationwide school malaria parasitaemia
survey in public primary schools, the United
Republic of Tanzania Frank Chacky1,2*† , Manuela Runge3,4*†, Susan F. Rumisha5, Pendael Machafuko2, Prosper Chaki6,
Julius J. Massaga5, Ally Mohamed1,2, Emilie Pothin3,4, Fabrizio Molteni2,3,4, Robert W. Snow7,8,
Christian Lengeler3,4 and Renata Mandike1,2 Abstract This article is distributed under the terms of the Creative Commons Attribution 4.0 International License
(http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium,
provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license,
and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/
publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Climate and seasonality
Th
l The climate in Tanzania is tropical with temperatures
between 25 and 31 °C during the hottest period (Novem-
ber to February) and between 15 and 20 °C in the cooler
period (May to August). In most parts of the country, the
temperature rarely falls below 20 °C, while in highland
areas the temperature ranges from 10 to 20 °C through-
out the year. There are two different rainfall seasons in
Tanzania: (1) from December to April in southern, south-
western, central and western parts of Tanzania, and (2)
from October to December and March to May in north-
ern and northern coastal parts of Tanzania [27]. A national, school, malaria parasitaemia survey (SMPS)
was conducted to close that data gap by increasing the
scope from children under 5 years to children 5–16 years
old and increase and complement the power of the popu-
lation surveys through increased sites and sample size. The SMPS was designed to allow estimates of malaria Geography and population Tanzania is among the East African countries, with a total
area of 945,000 sq. km. Mainland Tanzania is divided into
regions, councils, wards, and villages. Tanzania is mostly
rural, with around 28% of the population living in urban
councils, while 10% of the population live in Tanzania’s
largest city Dar es Salaam. According to the 2012 popula-
tion census, Tanzania has almost 45 million inhabitants,
with half of the population younger than 17 years [26]. In this context, school surveys have gained increased
attention for national surveillance, complementing
household surveys [10–12]. Schools are often well organ-
ized and easily accessible and provide the possibility of
collecting malariometric and control data from many
children in a short period of time and at a low cost com-
pared to TDHS/MIS, which are too costly for routine
surveillance. The importance of school survey data for
the planning of targeted interventions was demonstrated
by school surveys conducted in other African countries,
such as Kenya, Ethiopia, Uganda, Malawi, Côte d’Ivoire,
Democratic Republic of Congo, and The Gambia [10–
21], summarized by Brooker et al. [9]. Malaria in Tanzania Around 95% of the population is at risk of malaria [22]. Malaria transmission has been described as unstable,
seasonal in the arid central plateau, as stable seasonally
in the southern part (unimodal) and northern and west-
ern parts (bimodal), and stable throughout the year in
the coastal fringe, southern lowlands and the Lake Zone
[1]. In the past decade, there was an overall decrease in
malaria prevalence among children under the age of
5 years from 18.9 to 9.5% between 2008 and 2012 [23,
24]. However, the decline was not everywhere and even
increased in some parts of the country, leading to an
overall prevalence of 14% in 2016, observed in the 2015–
2016 TDHS-MIS survey [25]. Background prevalence and determine spatial and temporal risks of
Plasmodium falciparum transmission among public pri-
mary school-age pupils in mainland Tanzania. The objec-
tives were: (1) to determine the prevalence of malaria
among public primary school-enrolled pupils; (2) to
establish the spatial and temporal risks of P. falciparum
transmission across malaria-endemic councils; and, (3) to
determine the access and use of insecticide-treated bed
nets among school-age children. Tanzania is currently under epidemiological transition
from meso-endemic to hypo-endemic levels character-
ized by marked heterogeneity across and within regions
and/or councils [1]. This calls for an accurate and timely
estimate of the spatial–temporal distribution of malaria
transmission, malaria burden, and the impact of deployed
control interventions. Since 2000 it has been estimated
that the malaria burden has shown a marked decline in
sub-Saharan Africa, due to a large scale-up of control
interventions [2, 3]. Chacky et al. Malar J (2018) 17:452 Page 2 of 16 Methods Malaria prevalence among school-aged children in
Tanzania is under-researched and not well understood. To date, few data have been collected in older children
in a small-scale study, which have shown an increasing
proportion of the malaria burden in adolescents despite
transmission falls in the general population [4, 5]. In
addition, the observed high heterogeneity of malaria
transmission calls for the timely identification of popula-
tions and areas at greatest need for additional interven-
tions [6–8]. The major sources for malaria data are health
management information systems (HMIS) and large
household surveys, such as the malaria indicator surveys
(MIS) and Tanzania Demographic and Health Surveys
(TDHS). However, the HMIS captures only malaria cases
for those seeking care at health facilities, while house-
hold surveys which are conducted every 4–5 years have
been a useful tool to inform on the prevalence of malaria
in the country. Household surveys are logistically com-
plex, expensive, time consuming, and have limited scope
of sample size, whereby in Tanzania they focus only on
children under 5 years old [9]. Additionally, unsteady
funding and weak health systems hinder the timely and
reliable collection of data on malaria cases [6]. Education system and schools In Tanzania, in the year 2002, primary education had
been made compulsory to all eligible children [28]. According to the Ministry of Education and Vocational Chacky et al. Malar J (2018) 17:452 Page 3 of 16 Fig. 1 Locations of sampled schools (N = 537) coloured by survey date Fig. 1 Locations of sampled schools (N = 537) coloured by survey date 217 schools in 11 regions in May 2015; and, phase III
included 207 schools in 9 regions in October 2015
(Fig. 1). The sample was calculated and selected using a
multistage, stratified, proportional probability sampling
method. In each council, the number of children to be
tested was calculated based on the known population fig-
ures [26] and estimated mean population-weighted para-
site prevalence rate adjusted for children aged 2–10 years
(PfPR2–10) for 2010 [1]. The number of children to be
tested was then used to determine the number of schools
included in the survey in each council, assuming an aver-
age of 100 children per school. As a next step, the list of
wards in each council was confirmed and updated during
a national orientation meeting by local expert teams from
health and education sectors. The councils were strati-
fied according to the altitude, population density, demo-
graphic characteristics, and any other features available, Training data in 2015, there were 16,960 registered pri-
mary schools in mainland Tanzania, whereas 928 (5.5%)
of those were non-governmental [29]. The school enrol-
ment is described through intake and enrolment ratios,
whereas the National Intake Ratio (NIR) is the percent-
age of new enrolments of children of official school
entrance age over all children of official school entrance
age [30]. In 2014 it was estimated that the net enrolment
ratio ranged from 73% in Katavi region to nearly 100% in
Iringa region [30]. Training data in 2015, there were 16,960 registered pri-
mary schools in mainland Tanzania, whereas 928 (5.5%)
of those were non-governmental [29]. The school enrol-
ment is described through intake and enrolment ratios,
whereas the National Intake Ratio (NIR) is the percent-
age of new enrolments of children of official school
entrance age over all children of official school entrance
age [30]. In 2014 it was estimated that the net enrolment
ratio ranged from 73% in Katavi region to nearly 100% in
Iringa region [30]. School malaria parasitaemia survey (SMPS)
Survey design, procedures and study tools Proportionate stratifica-
tion was used to obtain the actual number of children
to be tested at each school. In each school, all seven pri-
mary school classes were included with an equal num-
ber of children in each class, balancing the sample for
gender groups. In phase III, only classes one to six were
included, due to the near end of the school term for chil-
dren in class seven. The selection of children in each class
was done separately for boys and girls, using a systematic
sampling procedure from a class register. A detailed flow-
chart about the sampling design can be found in Addi-
tional file 1: Fig. S1. Data entry Paper forms and RDT kits were transported to the head
office of the NMCP in Dar es Salaam for data entry and
storage. Data were single-entered by a group of trained
data entry clerks, using EpiData (EpiData Association,
Denmark) templates in phases I and III, and Microsoft
Excel (Microsoft Corporation, Seattle, USA) in phase II,
under supervision of study investigators and a statisti-
cian. Entered data into Epi-data were exported into Excel
for daily simple quality checks. Daily quality checks were
performed by comparing entered data per data entry
clerk for data entry errors using the filter function in
Excel. After data entry, 20% of data from each data entry
clerk were validated by comparing entered data with
paper forms. The whole dataset was screened for suspi-
cious data entries such as invalid values, out of range or
missing values, following a checklist for each variable. Suspicious entries were recorded, and hard copies revis-
ited to diagnose the entries as erroneous with correction,
or as invalid. Ethical considerations
E hi
l
l Ethical clearance was given by the National Health
Research Ethics Committee of NIMR. Informed consent
for the survey was based on a passive, opt-out method of
parental permission. Schools were instructed to inform
the students and parents concerning the survey. It was
assumed that parents approved their children’s participa-
tion if they did not express their disapproval. i
g
An average of 8 days was used for data collection per
region, ranging from 4 to 15 days. Within councils the
data collection took on average 4 days, ranging from 1
to 11 days (Additional file 2: Fig. S2). In each surveyed
school, a total of 2 days was used: one day for planning,
sampling and consenting and a second day for interview-
ing and testing. Each council was given an additional
1 day for data management and reporting. To test the
children for malaria parasites, SD BIOLINE Malaria Ag
P.f/Pan rapid malaria tests were used (Standard Diagnos-
tics Inc., Republic of Korea). The individual test results
were recorded in the designated RDT register and in
the respective individual children’s questionnaire. Fur-
thermore, the questionnaire captured information on
household size, bed net ownership and use, school absen-
teeism and fevers during the 2 weeks prior the survey
and, reported education level of one parent was obtained
in phase II and III. Children with a positive test result
were treated with artemether–lumefantrine (ALu), as
recommended in the National Malaria Diagnosis and
Treatment Guidelines [32] and referred to the nearest
health facility when necessary. To confirm the consist-
ency of recording of malaria test results, 20% of the used
rapid diagnostic tests were reviewed by quality assurance
team and the results were compared with the recorded
test results on the paper forms. For wrongly recorded
test results, the amendment was handled at the study site
by drawing a single line through an incorrect value and
dated. School malaria parasitaemia survey (SMPS)
Survey design, procedures and study tools A cross-sectional, school, malaria parasitaemia survey
was conducted in three phases: phase I which was con-
sidered a pilot phase, included 113 schools in 5 regions
between August and September 2014; phase II included Chacky et al. Malar J (2018) 17:452 Page 4 of 16 Page 4 of 16 such as urban/rural. The number of strata was equal to
the pre-defined number of schools, aiming to select one
school within each stratum. The stratification was done
to ensure that the study design captured the heterogene-
ity of malaria transmission at sub-council level. In each
stratum one ward and subsequently one village were ran-
domly selected, using a list of the administrative units
obtained from the National Bureau of Statistics Tanzania
[31]. Thus, the probability of a school to be selected was
dependent on the number of wards within the strata and
the number of villages within the selected ward. In case
there was more than one school located in a village, the
school was randomly selected. Proportionate stratifica-
tion was used to obtain the actual number of children
to be tested at each school. In each school, all seven pri-
mary school classes were included with an equal num-
ber of children in each class, balancing the sample for
gender groups. In phase III, only classes one to six were
included, due to the near end of the school term for chil-
dren in class seven. The selection of children in each class
was done separately for boys and girls, using a systematic
sampling procedure from a class register. A detailed flow-
chart about the sampling design can be found in Addi-
tional file 1: Fig. S1. such as urban/rural. The number of strata was equal to
the pre-defined number of schools, aiming to select one
school within each stratum. The stratification was done
to ensure that the study design captured the heterogene-
ity of malaria transmission at sub-council level. In each
stratum one ward and subsequently one village were ran-
domly selected, using a list of the administrative units
obtained from the National Bureau of Statistics Tanzania
[31]. Thus, the probability of a school to be selected was
dependent on the number of wards within the strata and
the number of villages within the selected ward. In case
there was more than one school located in a village, the
school was randomly selected. Survey staff and training Data collection was carried out by council teams, includ-
ing the malaria focal person, two laboratory techni-
cians, one education officer, and one teacher from each
surveyed school. Regional teams, including the regional
malaria focal person and a national supervisor from the
National Malaria Control Programme (NMCP), National
Institute for Medical Research (NIMR) or Ifakara Health
Institute (IHI), supervised data collection at regional
level and visited one council team per day to assess the
performance of malaria testing, assess data quality issues
and individuals’ interview. All teams participated in a
national 3-day training directly before the start of the
data collection. Data analysish The data were analysed in STATA version 14 [33] and
maps were created using QGIS [34]. Descriptive statistics
were done for all variables. Prevalence and proportions,
together with their 95% confidence intervals, were calcu-
lated, adjusted for clustering effect of children between Chacky et al. Malar J (2018) 17:452 Page 5 of 16 schools. Chi square tests were used to compare charac-
teristics of children and schools with the outcome vari-
ables “malaria infection” and “bed net use”. schools. Chi square tests were used to compare charac-
teristics of children and schools with the outcome vari-
ables “malaria infection” and “bed net use”. Population data were obtained from the national pop-
ulation census 2012, available from the National Bureau
of Statistics (NBS) Tanzania [42], and gridded popula-
tion densities from WorldPop [43]. Bed net use for a
child was defined as a binary variable stated as “yes” if
a child reported general use of net regardless of bed net
ownership and “no” otherwise. Malaria infection was
defined as “yes” for a positive RDT case regardless of
the malaria infection type and “no” if no infection was
detected. Variable definitions and sourcesh The age range was categorized into three groups: 5 to
under 9 years, 9 to 12 years, and older than 12 years. Councils were classified as urban or rural based on the
type of council. Municipals, township authorities and
city councils were classified as urban, remaining coun-
cils were classified as rural. Transmission zones were
classified according to the categories used in the most
recent epidemiological profile and based on the pre-
dicted mean PfPR2–10 from 2010, adjusted for the ages
2–10 years [1]. The following categories were created:
low stable (PfPR2–10 < 1%), hypo-endemic 1 (PfPR2–10 1
to < 5%), hypo-endemic 2 (PfPR2–10 5 to < 10%), meso-
endemic (PfPR2–10 10 to < 50%), and hyper-holo-endemic
(PfPR2–10 ≥ 50%). Bed net ownership was defined as hav-
ing at least one mosquito bed net in a household. Values
presented on malaria infection type [e.g. P. falciparum,
other Plasmodium species or Pf and pan infection (when
control and other two lines appear)], were based on the
RDT designated register dataset (N = 49,169), which
included slightly more children than the questionnaire
dataset (N = 49,113). The altitude was extracted for
the geo-location of the schools, using data from Shut-
tle Radar Topography Mission [35], downloaded from
WorldClim [36, 37]. The altitude was categorized into:
below 750 m, 750–1250 m, 1250–1750 m, and above
1750 m. Further environmental variables added were:
ecozone and temperature suitability index (TSI). The
ecozones were classified by the Food and Agriculture
Organization (FAO) as tropical rain forest, tropical moist
deciduous forest, tropical dry forest, tropical shrubland,
and tropical mountain system [38]. The temperature suit-
ability index is a relative measure of the impact of the
temperature on vectorial capacity (number of infectious
mosquitoes), ranging from 0 to 1 [39]. The TSI raster file
for 2015 was downloaded from the Malaria Atlas Project
at the University of Oxford [40] and mean values were
extracted per council using R version 3.3.1 [41]. Multilevel mixed‑effects logistic regression analysisl f
To assess the influence of risk factors for malaria infec-
tion and bed net use, multilevel mixed-effects logistic
regression models were fitted for each outcome sepa-
rately. Data were assumed to be clustered at council and
schools; hence, these were included in the model as ran-
dom effects. Variable selection was based on available
environmental, socio-economic and individual covari-
ates, which were previously associated with malaria, such
as annual rainfall, temperature and vegetation index,
TSI, altitude, type of council, reported education of par-
ents, gender, age, fever in the 2 weeks prior to survey. In
addition, log-likelihood ratio tests of bivariate models,
with a cut-off at a significance level of 0.01 were used. In
the model with outcome “malaria infection” the follow-
ing interaction terms were assessed: gender and bed net
use, age and bed net use, reported parental education
level and bed net use, reported parental education level
and urban area. Interaction terms were included at a sig-
nificance level of 0.05. The final set of selected variables,
which was used in both models included: gender, age, bed
net use or RDT result, reported parental education, type
of council, school-point altitude, geographical zone, and
school-point TSI. Due to limited available variables in
phase I, two multivariable models were calculated: model
I included data from all regions but had fewer covariates
(N = 47,157; 96%), whereas model II excluded data from
phase I regions but more covariates (N = 30,715; 62.5%)
(Table 1). Table 1 Multivariable regression models used in the present analysis
Model I
Model II
Regions
All
Excluding phase I data: Kigoma, Mtwara, Lindi, Ruvuma, Dar es Salaam
Number of observations
47,157 (96%)
30,715 (62.5%)
Variables
Included
Gender, age, bed net use or RDT result, type of
council, altitude, geographic zone, TSI
Gender, age, reported parental education, bed net use or RDT result,
type of council, altitude, eco-zone, geographic zone, TSI
Excluded
Reported parental education, eco-zone
Random effects
Council, school
Council, school Table 1 Multivariable regression models used in the present analysis Table 1 Multivariable regression models used in the present analysis Chacky et al. Malar J (2018) 17:452 Page 6 of 16 Table 2 Number of included councils, schools and children, by transmission area
* Source: Epidemiological profile Tanzania, 2013 [1]; ** no data for Mafia council in Pwani Region
Councils
Schools
Children
Total pop. Malaria infections
l In total, 10,627 children (21.6%) tested positive for
malaria (95% CI 19.5–23.9) (Table 3). Out of those, 6840
children (65%) had a mono-infection with P. falcipa-
rum, 3582 children (34.1%) had a malaria infection with
P. falciparum and a non-falciparum parasite (Plasmo-
dium vivax, Plasmodium ovale or Plasmodium malar-
iae) infection and 93 children (0.9%) were only infected
with a non-falciparum parasite. Furthermore, from the
positive-tested children, 2316 children (21.8%) reported
to have been diagnosed with malaria in the 2 weeks pre-
ceding the survey, and 2153 children (20.3%) received
treatment. Slightly more boys than girls were tested
positive [N = 5606 (23%, 95% CI 21–25.5%) vs N = 4940
(20%, 95% CI 18–22.2%)]. The malaria prevalence was
18.2% in children 5 to < 9 years (95% CI 16.1–20.5%),
21.4% in children aged 9–12 years (95% CI 19.2–23.8%)
and 23.3% in children older than 12 years (95% CI 21.2–
25.7%). The malaria prevalence was highest in children
reported parents with no school education (28.8%, 95%
CI 24.4–33.5%), compared to children with reported par-
ents with primary (22.0%, 95% CI 19.4–24.8%), reported
parents with secondary (17.8%, 95% CI 15.1–20.9%), and
reported parents to have higher education (9.9%, 95% CI
7.0–13.9%). In meso-endemic, and hyper-holo-endemic
councils almost one-third of the children were tested
malaria positive (31.4%, 95% CI 28.1–34.9%, and 29.9%,
95% CI 25.1–35.3%, respectively), in hypo-endemic
councils only 10% were tested positive (10.9%, 95% CI
8.3–14.1%), and in low stable endemic councils fewer
than 2% were tested positive (1.4%, 95% CI 0.5–3.7%)
(Fig. 2). The malaria prevalence slightly increased by age Multilevel mixed‑effects logistic regression analysisl (2010)
*, %
N
Children per council
N
Children per school
N
%
Mean
Min
Max
%
Mean
Min
Max
Transmission zone*
Low stable
30
253.5
111
546
88
86.4
44
166
7606
15.5
25.4
Hypo-endemic 1
40
296.1
64
501
130
91.1
42
157
11,845
24.1
20.4
Hypo-endemic 2
27
303.3
97
653
91
90
44
199
8188
16.7
13.3
Meso-endemic
67
316.5
89
1109
224
94.7
44
165
21,208
43.2
38.4
Hyper-holo-endemic
1
146
146
146
2
73
73
73
146
0.3
2.3
Missing**
1
120
120
120
2
60
60
60
120
0.2
–
Total
166
295.9
64
1109
537
91.5
42
199
49,113
100
100 Table 2 Number of included councils, schools and children, by transmission area more children reported to sleep under a bed net than
in rural councils (82.20%, 95% CI 78.7–85.2% vs 66.50%,
95% CI 64.1–68.8%) (Table 3). Results Overall 49,113 children in 537 schools were tested for
malaria and interviewed. On average 91 children were
sampled in each school, ranging from 44 to 199 children. Regarding the transmission zones, 7606 (15.5%) chil-
dren were sampled in councils classified as low stable,
11,845 (24.1%) in hypo-endemic 1; 8188 (16.7%) in hypo-
endemic 2; 21,208 (43.3%) in meso-endemic; and, 146
(0.3%) in hyper-holo-endemic (Table 2). Characteristics of sample Malaria test results were available for 49,102 (99.9%)
children and bed net use information was available for
47,800 children (97.3%). Majority of the children were
sampled in rural councils (80%). The mean altitude
was 1016 m (inter-quartile-range 615–1351 m) and the
mean TSI was 0.4 (inter-quartile-range 0.22–0.57). The
sample included as many boys as girls (boys 49.2%), and
the children were on average 11 years old (range 4–20;
inter-quartile-range 9–13 years) (Additional file 3: Fig. S3). Girls were found to be on average one year younger
than the boys. Majority of the children reported hav-
ing parents with primary school education (N = 23,445;
72%), while around 10% of the children reported having
parents who had never been to school (N = 3736; 11.6%). In the 2 weeks prior to the survey, 17,272 (35.7%) chil-
dren were absent due to sickness and 15,709 (32.7%) had
a fever; of those, 8383 (53.4%) children went to a health
facility for treatment and reported to be diagnosed with
malaria. Almost all the children diagnosed with malaria
had received treatment (N = 8155, 97.7%), while 1421
(14.8%) children received treatment but were not diag-
nosed with malaria. In total, 41,914 (89.5%) children
reported having at least one bed net at home, while the
bed net use was 69.6%. In urban councils significantly, Chacky et al. Characteristics of sample Malar J (2018) 17:452 Page 7 of 16 Table 3 Sample characteristics and risk factors for malaria infection and bed net use in school children in Tanzania
* No data for children sampled in phase I
Total children
Malaria (N = 49,102)
Bed net use (N = 47,800)
Children tested positive
Children sleeping under bed net
N
%
n
%
95% CI
n
%
95% CI
Total
49,113
100
10,627
22
(19.6–23.9)
33,284
70
(67.6–71.6)
Gender
Male
24,205
49.3
5606
23.2
(21.0–25.5)
16,077
68.3
(66.1–70.3)
Female
24,681
50.3
4940
20.0
(18.0–22.2)
17,078
71.0
(68.9–73)
Missing
227
0.5
81
35.7
–
129
63.9
–
Age
< 9
9075
18.5
1651
18.2
(16.1–20.5)
5974
68.8
(66.3–71.2)
9–12
18,892
38.5
4045
21.4
(19.2–23.8)
13,106
71.2
(69.2–73.3)
> 12
20,730
42.2
4847
23.4
(21.2–25.7)
13,957
68.6
(66.3–70.8)
Missing
416
0.9
84
20.2
–
247
62.7
–
Parental education
No school
3736
7.6
1074
28.8
(24.4–33.5)
1930
54.7
(50.2–59.1)
Primary
23,445
47.7
5163
22.0
(19.4–24.8)
14,968
65.8
(63.1–68.3)
Secondary
4547
9.3
808
17.8
(15.1–20.9)
3430
77.5
(74.8–80)
Diploma or higher
525
1.1
52
9.9
(7.0–13.9)
415
80.7
(76.1–84.7)
Missing
5424
11.0
970
17.9
–
3617
69.7
–
Not interviewed*
11,436
23.3
2560
22.4
–
8924
78.4
–
Bed net use
No
14,516
29.6
2943
20.3
(17.8–23.0)
–
–
–
Yes
33,284
67.8
7270
21.9
(19.6–24.2)
–
–
–
Missing
1313
2.7
414
31.5
–
–
–
–
Zone
Eastern
6967
14.2
1269
18.2
(13.3–24.4)
5819
86.6
(83.4–89.3)
Western
4805
9.8
1449
30.2
(25.5–35.2)
3221
68.8
(64–73.2)
Southern
4002
8.1
1344
33.6
(27.2–40.8)
3255
81.6
(76.8–85.6)
Southern highlands
4116
8.4
495
12.0
(7.2–19.4)
2350
58.7
(52.4–64.7)
Southwest highlands
4867
9.9
846
17.4
(11.6–25.2)
2946
61.6
(55.3–67.5)
Central
5653
11.5
156
2.8
(1.6–4.6)
3077
55.1
(49.6–60.5)
Northern
6191
12.6
317
5.1
(3.1–8.4)
3360
55.6
(49.4–61.7)
Lake
12,512
25.5
4751
38.0
(33.6–42.5)
9256
77.2
(73.1–80.8)
Missing
0
0.0
0
0.0
–
0
0.0
–
Area
Urban
9708
19.8
588
6.1
(4.0–9.1)
7866
82.2
(78.7–85.2)
Rural
39,405
80.2
10,039
25.5
(23.1–28.0)
25,418
66.5
(64.1–68.8)
Missing
0
0.0
0
0.0
–
0
0.0
–
Eco-zone (tropical) *
Dry forest
7124
19.0
2247
31.5
(25.9–37.8)
5417
79.4
(74.7–83.4)
Moist decid. * No data for children sampled in phase I 1 If not otherwise stated, the described results refer to the multivariable
regression analysis model II. The results from the regression model I can be
obtained from Table 4. Characteristics of sample forest
5819
15.5
1816
31.2
(25.1–38)
3786
67.8
(62–73.1)
Mountain system
6053
16.1
723
11.9
(7.9–17.6)
3329
57.0
(50.3–63.5)
Rainforest
3331
8.9
1272
38.2
(28.6–48.7)
2664
82.8
(76.3–87.8)
Scrubland
15,216
40.5
2009
13.2
(10.3–16.7)
9078
61.2
(57.4–64.9)
Missing
11,570
23.6
2560
22.2
–
9010
77.9
–
Altitude (m)
< 750
13,228
26.9
3248
24.6
(20.7–28.9)
10,697
82.7
(80.1–85)
750–1250
18,901
38.5
5040
26.7
(23.3–30.4)
13,284
72.0
(68.8–75)
1250–1750
14,581
29.7
2335
16.0
(12.6–20.1)
8434
59.8
(56–63.4)
> 1750
2403
4.9
4
0.2
(0.1–0.5)
869
37.8
(30.5–45.7)
Missing
0
0.0
0
0.0
–
0
0.0
– cteristics and risk factors for malaria infection and bed net use in school children in Tanzania Table 3 Sample characteristics and risk factors for malaria infection and bed net use in school children in Tanzania Chacky et al. Malar J (2018) 17:452 Page 8 of 16 Fig. 2 Malaria prevalence, bed net use, school absenteeism, and recent fever of school children compared by transmission zone. Error bars indicate
95% CI adjusted for school clustering Fig. 2 Malaria prevalence, bed net use, school absenteeism, and recent fever of school children compared by transmission zone. Error bars indicate
95% CI adjusted for school clustering in all transmission zones (Additional file 4: Fig. S4). The
malaria prevalence was highest at altitudes below 750 m
and between 750 and 1250 m above sea level (24.6 and
26.7%) and lowest in areas 1750 m above sea level (0.2%),
(16.0% at 1250–1750 m). In rural councils, the malaria
prevalence was four times higher than in urban councils
(25.5%, 95% CI 23.1–28.0% vs 6.1%, 95% CI 4.0–9.1%)
(Table 3). Exceptions were urban councils of Geita TC,
Handeni TC, Kigoma MC and Masasi TC, with preva-
lence higher than the national average. 1.21; 95% CI 1.14–1.28). The association between
malaria infection and age was not significant, (children
9–12 years OR 1.03; 95% CI 0.92–1.15, and children
older than 12 years OR 1.05; 95% CI 0.95–1.17 compared
to children 5–9 years). The association between age and
bed net use was significant, but the difference in the odds
ratio was only marginal (OR 1.10 vs OR 1.12). Children
who reported to sleep under a bed net were less likely to
have malaria (OR 0.81; 95% CI 0.74–0.88) (Table 4). Council type In comparison to children living in the Central Zone,
children living in Northern and Southern Highlands
had lower odds of malaria infection, and children living
in the other zones had higher odds of malaria infection. For instance, children living in the Lake Zone had 24.4
times higher odds of malaria infection compared to chil-
dren living in the Central Zone (95% CI 10.4–57.1). Chil-
dren living in Southern Highlands and Northern Zones,
were less likely to sleep under a bed net (OR 1.57; 95% CI
0.78–3.15, and OR 0.72; 95% CI 0.40–1.30), while higher
odds ratios were found in children living in the Lake
Zone (OR 3.54; 95% CI 2.03–6.18), compared to children
living in the Central Zone. The odds of malaria infection
significantly decreased at higher altitudes, with an odds
ratio of 0.26 at 750–1250 m (95% CI 0.11–0.62), an odds
ratio of 0.25 at 1250–1750 m (95% CI 0.09–0.72) and an
odds ratio of 0.07 at altitudes higher than 1750 m (95% CI
0.01–0.47), compared to areas below 750 m. The odds for
children to sleep under a bed net decreased significantly
with increasing altitude (OR 0.43; 95% CI 0.25–0.74 at
750–1250 m, OR 0.27; 95% CI 0.14–0.53 at 1250–1750 m,
and OR 0.12; 95% CI 0.05–0.31 at 1750 m and higher,
compared to 750 m and lower). Living in urban councils was significantly associated with
lower odds of malaria infection, compared to living in
rural councils (OR 0.15, 95% CI 0.08–0.29). Children liv-
ing in urban councils were two times more likely to sleep
under a bed net than children living in rural councils (OR
1.92; 95% CI 1.25–2.94). Reported education of parentsh The error bars present
the 95% CI adjusted for school clustering and the dashed line presents the mean for all ages Reported education of parentsh Figure 3 shows the main indicators among school chil-
dren by age. Bed net use was high and close to the mean
for all ages until it declined at age 14 years. The percent-
age of children who reported to be absent from school
due to sickness in the 2 weeks preceding the survey was
higher in younger children and decreased after the age
of 11 years. The percentage of children who reported to
have had a fever went to a health facility and were diag-
nosed with malaria was higher in younger children and
started to decline after the age of 10 years. With increas-
ing age fewer children were diagnosed with malaria at a
health facility in the 2 weeks before the survey, while the
percentage of children tested positive for malaria at the
day of the survey showed an increasing trend with age
(Fig. 3). The reported education of the parents was strongly asso-
ciated with both malaria infection and bed net use, with
lower odds of malaria infection and higher odds of bed
net use in children associated with higher educated par-
ents. For malaria infection, the odds ratio in children with
parents with no school education was 1.18 (95% CI 1.06–
1.32), and in children with parents with secondary or
higher education 0.75 (95% CI 0.67–0.83), compared to
children with parents with primary education. The odds
ratios of bed net use increased with increasing education
of the parents. Children with non-educated parents had
an odds ratio of 0.69 (95% CI 0.63–0.76), with secondary
or higher education was 1.48 (95% CI 1.35–1.61), com-
pared to children reported to have parents with primary
education. Malaria infection, bed net use and associated risk factors
In the multivariable analysis1 girls had lower odds of
malaria infection than boys (OR 0.74; 95% CI 0.69–
0.79) and higher odds of sleeping under a bed net (OR Chacky et al. Malar J (2018) 17:452 Page 9 of 16 Fig. 3 Bed net use, measured malaria prevalence, school absenteeism, and history of sickness in previous 2 weeks, by age. The error bars present
the 95% CI adjusted for school clustering and the dashed line presents the mean for all ages Fig. 3 Bed net use, measured malaria prevalence, school absenteeism, and history of sickness in previous 2 weeks, by age. Eco‑zone For the risk of malaria infection as well as bed net use,
it made no significant difference in which eco-zone
the children were living, the chance of malaria infec-
tion was higher in dry forest (OR 2.06, 95% CI 1.42–
3.72), moist deciduous forest areas (OR 2.36, 95% CI
1.21–4.60), mountain system (OR 1.51, 95% CI 0.68–
3.38), and three times higher in rainforest areas (OR
3.01, 95% CI 1.41–6.40) compared to scrubland areas. Regarding bed net use, children in tropical rainforest
areas were more likely to use a bed net (OR 1.33; 95%
CI 0.76–2.31), while children in moist deciduous for-
est were least likely to use a bed net (OR 0.89; 95% CI
0.57–1.38), in comparison to tropical scrubland areas
(Table 4). Chacky et al. Malar J (2018) 17:452 Page 10 of 16 5%, in 27 between 5 and 10%, in 25 between 10 and 25%,
in 51 between 25 and 50%, and in 18 councils above 50%. Highest malaria prevalence was found in Geita, Pwani,
Mwanza, and Katavi (≥ 40%); lowest prevalence in Aru-
sha, Kilimanjaro, Manyara and Iringa (< 1%) (Fig. 4). Geographical distribution of observed malaria prevalence
The malaria prevalence was highest in the Lake Zone
(38%, 95% CI 33.5–42.4), Southern Zone (33%, 95% CI
26.8–40.5%), as well as Western Zone (30%, 95% CI 25.3–
35%), and lowest in Central (2.8%, 95% CI 1.4–4.2%),
and Northern zones (5.1%, 95% CI 2.5–7.7%). At council
level, the malaria prevalence ranged from 0 to 76.4%. Eco‑zone In
62 of the 166 councils, the malaria prevalence was below Between schools, the variation of the malaria preva-
lence was greatest in regions with a high mean preva-
lence, while the variation was marked by only a few Table 4 Multivariable analysis of the risk factors for malaria and bed net use in primary school children in Tanzania
Outcome/covariates
Univariable
Model I (N = 47,157)
Model II (N = 30,715)
Malaria
Malaria
Malaria
Bed net use
OR (95% CI)
p- value
OR (95% CI)
p-value
OR (95% CI)
p-value
OR (95% CI)
p-value
Gender
Male vs female
0.75 (0.71–0.79)
< 0.01
0.77 (0.72–0.81)
< 0.01
0.74 (0.69–0.79)
< 0.01
1.21 (1.14–1.28)
< 0.01
Age (years)
< 9 vs 10–12
1.06 (0.98–1.15)
0.11
1.05 (0.96–1.14)
0.36
1.03 (0.92–1.15)
0.55
1.1 (1.01–1.20)
0.02
< 9 vs > 12
1.09 (1.01–1.18)
1.06 (0.98–1.15)
1.05 (0.95–1.17)
1.12 (1.03–1.22)
Parental education
Primary vs no school
1.18 (1.06–1.31)
< 0.01
–
1.18 (1.06–1.32)
< 0.01
0.69 (0.63–0.76)
< 0.01
Primary vs secondary or
higher
0.74 (0.67–0.82)
–
0.75 (0.67–0.83)
1.48 (1.35–1.61)
Bed net use
No vs yes
0.76 (0.71–0.81)
< 0.01
0.76 (0.71–0.82)
< 0.01
0.81 (0.74–0.88)
< 0.01
–
Malaria
Negative vs positive
–
–
–
0.81 (0.74–0.88)
< 0.01
Area
Rural vs urban
0.14 (0.05–0.38)
< 0.01
0.12 (0.066–0.21)
< 0.01
0.15 (0.072–0.29)
< 0.01
1.92 (1.25–2.94)
< 0.01
Altitude (m)
< 750 vs 750–1250
0.25 (0.12–0.48)
< 0.01
0.18 (0.081–0.39)
< 0.01
0.26 (0.11–0.62)
0.02
0.43 (0.25–0.74)
0.01
< 750 vs 1250–1750
0.07 (0.04–0.15)
0.12 (0.045–0.31)
0.25 (0.088–0.72)
0.27 (0.14–0.53)
< 750 vs > 1750
0.002 (0.00–0.02)
0.02 (0.003–0.14)
0.07 (0.0093–0.47)
0.12 (0.049–0.31)
Temperature suitability index
TSI (2er intervals)
3.51 (2.80–4.39)
< 0.01
–
< 0.01
2.2 (1.56–3.09)
< 0.01
1 (0.81–1.24)
0.98
Zone
Central vs eastern
28.15 (7.68–103.20)
< 0.01
1.86 (0.60–5.72)
< 0.01
2.11 (0.63–7.01)
< 0.01
2.13 (0.96–4.71)
< 0.01
Central vs western
59.21 (15.40–228.10)
31.13 (11.6–83.4)
11.21 (3.23–39.0)
2.18 (0.91–5.19)
Central vs southern
72.58 (17.70–296.90)
2.79 (0.83–9.33)
Central vs southern high-
lands
2.13 (0.53–8.53)
2.64 (0.91–7.67)
0.41 (0.092–1.85)
0.71 (0.33–1.52)
Central vs southwest
highlands
10.33 (2.82–37.80)
6.7 (2.52–17.9)
3.19 (1.06–9.60)
1.57 (0.78–3.15)
Central vs northern
1.05 (0.29–3.77)
0.47 (0.17–1.31)
0.43 (0.15–1.19)
0.72 (0.40–1.30)
Central vs lake
69.76 (23.0–211.9)
40.97 (18.0–93.4)
24.43 (10.4–57.1)
3.54 (2.03–6.18)
Eco-zone (tropical)
Scrubland vs dry forest
4.92 (2.44–9.94)
< 0.01
–
2.06 (1.14–3.72)
0.22
0.93 (0.61–1.41)
0.81
Scrubland vs moist deci. Eco‑zone forest
4.98 (2.31–10.73)
–
2.36 (1.21–4.60)
0.89 (0.57–1.38)
Scrubland vs mountain
system
0.56 (0.23–1.34)
–
1.51 (0.68–3.38)
1.01 (0.64–1.60)
Scrubland vs rainforest
7.63 (2.90–20.08)
–
3.01 (1.41–6.40)
1.33 (0.76–2.31) Table 4 Multivariable analysis of the risk factors for malaria and bed net use in primary school child ble analysis of the risk factors for malaria and bed net use in primary school children in Tanzania Chacky et al. Malar J (2018) 17:452 Page 11 of 16 Fig. 4 Geographical distribution of the observed mean malaria prevalence among school children per council. The prevalence shown is the
unadjusted observed prevalence, measured in different times of the year Fig. 4 Geographical distribution of the observed mean malaria prevalence among school children per council. The prevalence shown is the
unadjusted observed prevalence, measured in different times of the year prevalence in the Lake Zone was surprising. One possible
explanation could be the El Ninõ Southern Oscillation
(ENSO) unstable climate conditions, which had caused
‘dramatic malaria outbreaks’ before [1]. outlying schools in regions with a low mean prevalence
(Fig. 5). The geographical pattern of the malaria preva-
lence aggregated by councils and the heterogeneity
among prevalence in schools within regions separated by
survey phase is shown in Additional file 5: Fig. S5. Discussion The percentage of reported bed net ownership in the
children’s household was much higher than the reported
percentages in the TDHS-MIS (89 vs 65%). However, the
trend among regions was similar, with lower percentages
in the middle part of Tanzania and higher percentages in
regions in the Lake Zone and Southern Zone of Tanza-
nia. The high reported bed net ownership among school
children could likely be influenced by recall, report-
ing or interviewer bias, leading to overestimated values,
although it was found that school children give reliable
answers about bed net coverage in their community [21]. The percentage of children generally sleeping under a bed
net was higher than the percentage for children under
the age of 5 years, who slept under a bed net the previous
night (69.9 vs 54.5%) [25]. This is different from previ-
ous findings that school children were less likely to sleep
under a bed net than other age groups [46]. The dispar-
ity may be explained by the differences in the terms used, Almost 50,000 children from 537 public primary schools
across all transmission zones in Tanzania were inter-
viewed and tested for malaria infection. The nationwide
malaria prevalence was 21.6%, marked by high variation
across transmission zones, and administrative borders. The malaria prevalence in Tanzania was low in the “mid-
dle corridor”, ranging from Arusha and Kilimanjaro in the
northeast to Njombe and Rukwa in the southwest of Tan-
zania, and high in the northwest (mainly Lake Zone) and
in the southeast lowlands of Tanzania. The geographi-
cal pattern was similar to the previously described geo-
graphical pattern of malaria prevalence and vulnerability
[1, 44, 45]. Compared to the TDHS-MIS 2015–2016, the
national and regional malaria prevalence among school
children was higher than the prevalence among children
under the age of 5 years (21.6 vs 14%) [25]. Although the
trend across the regions was similar, the especially high Chacky et al. Malar J (2018) 17:452 Page 12 of 16 Fig. 5 Distribution of malaria prevalence in schools by region. The regions are sorted by regional mean malaria prevalence. Geita had the highest
prevalence (53.7%) and Arusha the lowest prevalence (< 0.1%). The grey box visualizes the interquartile range (25–75%) of the school prevalence
within each region. The horizontal line within the grey box is the median of the school prevalence distribution. The spikes mark the lowest and
highest quartile. Statistical analysis y
Considering the variety of putative risk factors for
malaria and bed net use, the regression models were not
fully adjusted and in future analysis, it would be inter-
esting to include risk factors such as distance to nearest
health facility, distance to nearest water body, popula-
tion density, housing conditions and/or socio-economic
status. Also, the model included TSI and altitude, which
were correlated but remained significant in the multi-
variable log-likelihood ratio test. Children, excluded in
model I, adjusted for education and eco-zone had signifi-
cantly higher reported bed net use and were on average
younger than the children included in the model. One
possible reason for the higher percentage of bed net use
in the excluded children would be the school net distri-
bution campaign in Southern Tanzania (Lindi, Mtwara,
Ruvuma) in 2013 [53]. This would be supported by the
reported higher percentages of obtained bed net through
schools in Lindi and Mtwara region in the TDHS-MIS
[25]. Further analysis could use stratification by region or
transmission zone, geo-spatial analysis or geographically Discussion The points present outlying schools with prevalence higher or lower most of the rest of the prevalence (1.5 times the interquartile
range above the upper quartile/below the lower quartile) Fig. 5 Distribution of malaria prevalence in schools by region. The regions are sorted by regional mean malaria prevalence. Geita had the highest
prevalence (53.7%) and Arusha the lowest prevalence (< 0.1%). The grey box visualizes the interquartile range (25–75%) of the school prevalence
within each region. The horizontal line within the grey box is the median of the school prevalence distribution. The spikes mark the lowest and
highest quartile. The points present outlying schools with prevalence higher or lower most of the rest of the prevalence (1.5 times the interquartile
range above the upper quartile/below the lower quartile) whereas “general” bed net use is much broader than bed
net uses the previous night, leading to an overestima-
tion of bed net use among school children. While on the
other hand, bed net use among children under the age of
5 decreased since the previous MIS survey in 2011–2012
[24, 25] which is attributed to variations in distribution
campaigns. Bed net use was higher in urban than in rural
councils, which is similar to the findings in the TDHS-
MIS. Moreover, bed net ownership and use were higher
among children with higher educated parents, which is
concordant with the TDHS-MIS findings [24, 25], assum-
ing parental education and socio-economic status of the
household to be the same indicator as with findings from
a survey in Uganda [47]. [51]. However, the association between malaria and age
highly depends on the transmission intensity among
many other factors [48, 52]. The lack of association in the
univariable model could be due to the fact that it was not
distinguished between low and high transmission zones. Malaria and its risk factorsh The small increase in the malaria prevalence by age at all
transmission zones, as well as the lack of significant asso-
ciation between age and malaria, might be surprising. Whilst the proportion of children with malaria slightly
increased by age in all transmission zones, it would have
been expected that younger children have a higher preva-
lence in high transmission areas and that older children
would have a higher prevalence in low transmission
zones [48]. A significant association between age and
malaria was found in Ghana [49] and for a high transmis-
sion zone in Kenya [19, 50], whereas no association was
found in a school survey in Ethiopia and Côte d’Ivoire Chacky et al. Malar J (2018) 17:452 Chacky et al. Malar J (2018) 17:452 Page 13 of 16 Page 13 of 16 weighted logistic regression to assess the spatial correla-
tion and its association with risk factors of malaria or bed
net use. enrolment rates were lowest in Kigoma, Katavi and Man-
yara regions (72.9, 73.6 and 79.8%, respectively), and oth-
erwise mostly above 90% [30]. School absenteeism caused
by malaria infection could have introduced a ‘healthy
child effect’. This would lead to an underestimation of
malaria prevalence, especially in low transmission areas,
where children would be more likely to show symptoms
and stay at home when infected [10, 11]. In Tanzania,
there is not much known about how well school preva-
lence reflects community prevalence, although a study in
Kenya found that data obtained in schools may reflect the
community [10, 59]. Moreover, the use of school survey
data for sentinel surveillance at community level could be
validated in further research. Seasonality
Th
S
S y
The SMPS, as all cross-sectional surveys, captures
malaria infection prevalence only at a certain time
point and seasonal variations are inevitable. In the mid-
dle corridor, with low malaria prevalence, the survey
was conducted during the end of wet season in most
of the councils, while around the Lake Zone and in the
southern parts of the country, with high malaria preva-
lence, the survey was conducted during the beginning of
the short rains and dry seasons. The authors acknowl-
edge that in Tanzania there is no quantifiable way to
adequately and entirely adjust for differences in timing
of the survey with expected prevalence rate, and this is
a challenge even in most sophisticated mapping models
or malaria prevalence. However, most school children
(5–16 years), including those in this study, carry asymp-
tomatic infections [16], which are less likely to be treated;
thus, it has been documented that school children tend
to harbour such infection for long periods, over 5 months
[60, 61], potentially reducing the influence of seasonality
on malaria infections in this age group. Future alternative
sampling methods might include surveys at the begin-
ning of every term, or more frequent surveys throughout
the year, similar to the approach of rolling malaria indica-
tor surveys. Validity of children interviews Self-reported values are likely to differ from the truth,
depending on interviewer and respondent characteris-
tics, and face-to-face interviews are more likely to lead to
answers influenced by social expectations [57, 58]. Since
interviews were not standardized, interviewer bias might
have varying influence on results. However, considering
the large scale and sample size, it is less likely that this
had an impact on the results at council level. Malaria testing Malaria rapid diagnostic tests were used to test children
for malaria, which may have led to an overestimation in
high transmission areas since the RDT were found to
be limited in their ability to distinguish between “active
and resolved infections” [54]. Also, although not further
investigated in the present analysis, the number of pos-
itive-tested children may be biased by the proportion of
children who had taken anti-malarial treatment recently
before the day of testing. In future, this information could
be of use for evaluation of the performance of malaria
diagnosis and treatment at sub-regional level among
school children. In low transmission areas, the use of
RDTs may underestimate true prevalence, since very low
parasitaemia, which lies below the detection level of the
rapid diagnostic tests, were missed [54, 55]. It has been
recommended to use molecular detection tools in low
transmission areas [55]. This would enable school sur-
veys to track down remaining human parasite reservoirs,
which would contribute to an improved evaluation of
the progress of malaria elimination in low transmission
zones [56]. Moreover, it would be of interest to evaluate
the use of schools in addition to health facilities for active
case detection in the community, to identify remaining
parasite reservoirs in low transmission zones and to iden-
tify hotspots within the catchment area of the schools. Funding g
Phase I of the School Malaria Parasitaemia Survey used funds from the
Department of International Development, UK through the University of
Oxford as part of the INFORM project. Phase II and III of the School Malaria
Parasitaemia Survey used funds from the Global Fund for ATM, Grant Number:
TNZ-M-MOFEA. Additional file 5: Fig. S5. A Map of the observed prevalence among
schools aggregated per council, separated by survey phase. B Boxplot
showing the distribution of observed prevalence among schools per
region, sorted by amount of rainfall in the 3 months preceding the survey
and separated by survey phase. Acknowledgements This study was conducted by the National Malaria Control Program, Ministry
of Health, Community Development, Gender, Elderly, and Children, Tanzania. The authors would like to thank Sigsbert Mkude, Witness Mchwampaka, Erasto
Kazyoba, Fidelis Mgohamwende, and Abdallah Kajuna for their support in
designing and conducting the study, Amanda Ross for helpful conversations,
Tom Smith for scientific support. RWS is supported as a Principal Research
Fellow by the Wellcome Trust, UK (# 103,602). Competing interests Competing interests
The authors declare that they have no competing interests. The authors declare that they have no competing interests. Ethics approval and consent to participate Ethical clearance was given by the National Health Research Ethics Committee
of the National Institute for Medical Research (NIMR). Informed consent for
the survey was based on a passive, opt-out method of parental permission. Schools were instructed to inform the students and parents concerning the
exercise. It was assumed that parents who did not express their disapproval
for the survey, approved children’s participation. Advantages of SMPS compared to MISh This SMPS, which is powered with adequate sample size
to provide malaria prevalence estimates at council and
sub-council levels, provides a complementary approach
to malaria surveillance and parasitological monitoring in
a short period of time alongside with other national rep-
resentative surveys, such as Tanzania DHS and MIS. The
nationwide school survey is the first survey in Tanzania
describing the malaria risk among school children and
the geographical trend. This is of importance for malaria
control and elimination, as malaria infections in school
children are often asymptomatic, contributing to malaria The data were analysed without adding sampling weights
to account for the varying probability of selection at
sub-council level. Nevertheless, with respect to repre-
sentativeness, the study design included a population-
weighted selection of council and stratum. In addition,
the lack of available data at sub-council level to determine
sampling weights would have provided limited additional
accuracy to the results. Another limitation of the repre-
sentativeness would be school absenteeism at day of the
survey, drop out and enrolment rates in schools. Accord-
ing to a national educational survey conducted in 2013, Chacky et al. Malar J (2018) 17:452 Page 14 of 16 4 University of Basel, Basel, Switzerland. 5 National Institute for Medical
Research, Dar es Salaam, Tanzania. 6 Ifakara Health Institute, Dar es Salaam,
Tanzania. 7 KEMRI-Welcome Trust Research Programme, Nairobi, Kenya. 8 Cen-
tre for Tropical Medicine and Global Health, Nuffield Department of Clinical
Medicine, University of Oxford, Oxford, UK. transmission in the community as ‘hidden’ parasite res-
ervoirs [16]. The SMPS costs US$10 per test performed
compared to MIS, which costs an average of US$410 per
test performed. However, apart from malaria testing,
MIS also captures household information such as cover-
age indicators including ownership and use of bed net,
use of intermittent preventive treatment (Sulfadoxine
Pyrimethamine) during pregnancy, and social and behav-
iour change communication [26]. References 1. National Malaria Control Programme (Tanzania), WHO (Tanzania),
Ifakara Health Institute (Tanzania), KEMRI-Wellcome Trust (Kenya). An
epidemiological profile of malaria and its control in mainland Tanzania. Report funded by Roll Back Malaria and Department for International
Development-UK; 2013. p. 152. 1. National Malaria Control Programme (Tanzania), WHO (Tanzania),
Ifakara Health Institute (Tanzania), KEMRI-Wellcome Trust (Kenya). An
epidemiological profile of malaria and its control in mainland Tanzania. Report funded by Roll Back Malaria and Department for International
Development-UK; 2013. p. 152. Authors’ contributions FC, RM, SFR, PC and FM were responsible for study design, survey tools and
fieldwork supervision. FC, SFR, PC, FM, PM and MR participated in the data
collection. FC, PM, PC and SR supervised data collection. FC provided logistic
guidance. FC, RM, and AM provided administrative guidance. MR and PM
cleaned the data. MR analysed the data and wrote the draft manuscript. FC,
MR, SFR, and PM supervised data entry processes and interpretation of find-
ings. FM, CL, EP, FC, PM, SFR contributed to the interpretation of findings. FM,
CL and EP provided technical advice to data analysis. FC, PM, SFR, EP, FM, AM,
JJM, RWS and CL provided technical inputs to the manuscript. EP, SFR, FM,
RWS and CL provided scientific expertise. All authors read and approved the
final manuscript. 2. Bhatt S, Weiss DJ, Cameron E, Bisanzio D, Mappin B, Dalrymple U, et al. The effect of malaria control on Plasmodium falciparum in Africa between
2000 and 2015. Nature. 2015;526:207–11. 3. WHO. World Malaria Report 2015. Geneva: World Health Organization;
2015. 4. Mazigo HD, Rumisha SF, Chiduo MG, Bwana VM, Mboera LEG. Malaria
among rice farming communities in Kilangali village, Kilosa district,
Central Tanzania: prevalence, intensity and associated factors. Infect Dis
Poverty. 2017;6:101. 5. Nzobo BJ, Ngasala BE, Kihamia CM. Prevalence of asymptomatic malaria
infection and use of different malaria control measures among primary
school children in Morogoro Municipality, Tanzania. Malar J. 2015;14:491. Additional files Additional file 1: Fig. S1. Flow chart of sampling design. Additional file 2: Fig. S2. Duration of data collection in days per survey
phase and district. Additional file 3: Fig. S3. Age histogram. Additional file 4: Fig. S4. Malaria prevalence per age by transmission
zone. Additional file 5: Fig. S5. A Map of the observed prevalence among
schools aggregated per council, separated by survey phase. B Boxplot
showing the distribution of observed prevalence among schools per
region, sorted by amount of rainfall in the 3 months preceding the survey
and separated by survey phase. Additional file 1: Fig. S1. Flow chart of sampling design. Additional file 2: Fig. S2. Duration of data collection in days per survey
phase and district. Additional file 3: Fig. S3. Age histogram. Additional file 4: Fig. S4. Malaria prevalence per age by transmission
zone. Additional file 5: Fig. S5. A Map of the observed prevalence among
schools aggregated per council, separated by survey phase. B Boxplot
showing the distribution of observed prevalence among schools per
region, sorted by amount of rainfall in the 3 months preceding the survey
and separated by survey phase. Abbreviations ALu: artemether–lumefantrine; HMIS: Health Management Information Sys-
tem; MIS: Malaria Indicator Surveys; RDT: malaria rapid diagnostic test; NMCP:
National Malaria Control Programme; SMPS: School Malaria Parasitaemia
Survey; TDHS: Tanzania Demographic and Health Surveys; TSI: temperature
suitability index. Conclusionh The observed overall malaria prevalence of 21.6% rang-
ing from < 0.01 (Arusha, Manyara) to 53.6% (Geita)
among regions indicates similar malaria heterogeneity
and patterns as reported in the national surveys (TDHS/
MIS). Findings of this survey are useful for updating the
malaria epidemiological profile and for stratification
of the malaria transmission by region, council and age
groups, which is essential for guiding resource allocation
and prioritizing future malaria interventions. Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in pub-
lished maps and institutional affiliations. Received: 26 July 2018 Accepted: 29 November 2018 Received: 26 July 2018 Accepted: 29 November 2018 Availability of data and materials The datasets generated and analysed during the current study are not publicly
available but are available from the principal investigator on a reasonable
request. 5.
Nzobo BJ, Ngasala BE, Kihamia CM. Prevalence of asymptomatic malaria
infection and use of different malaria control measures among primary
school children in Morogoro Municipality, Tanzania. Malar J. 2015;14:491. Author details
1 1 Ministry of Health, Community Development, Gender, Elderly, and Children,
Dodoma, Tanzania. 2 National Malaria Control Programme, Dar es Salaam,
Tanzania. 3 Swiss Tropical and Public Health Institute, Basel, Switzerland. Chacky et al. Malar J (2018) 17:452 Chacky et al. Malar J (2018) 17:452 Page 15 of 16 Indicator Survey (TDHS-MIS) 2015-2016. Dar es Salaam. 2016. http://dhspr
ogram.com/pubs/pdf/FR321/FR321.pdf. Accessed 3 Jul 2017. 6. Hemingway J, Shretta R, Wells TNC, Bell D, Djimdé AA, Achee N, et al. Tools
and strategies for malaria control and elimination: what do we need to
achieve a grand convergence in malaria? PLoS Biol. 2016;14:e1002380. Indicator Survey (TDHS-MIS) 2015-2016. Dar es Salaam. 2016. http://dhspr
ogram.com/pubs/pdf/FR321/FR321.pdf. Accessed 3 Jul 2017. 26. National Bureau of Statistics (Tanzania), Tanzania, Office of Chief Govern-
ment Statistician (Zanzibar). 2012 Population and Housing Census. Dar es
Salaam.2013. 7. Tanner M, Greenwood B, Whitty CJM, Ansah EK, Price RN, Dondorp AM,
et al. Malaria eradication and elimination: views on how to translate a
vision into reality. BMC Med. 2015;13:167. 27. Government Portal Content Committee. Tanzania Government Portal—
Tanzania Profile. 2013. http://www.tanzania.go.tz. Accessed 7 Nov 2015. 8. WHO. Global Technical Strategy for Malaria 2016–2030. Geneva: World
Health Organization; 2015. 28. Ministry of Education and Vocational Training. Education for All 2015
National Review Report: United Republic of Tanzania Mainland. Ministry
of Education and Vocational Training: UNESCO; 2014. 9. Brooker S, Kolaczinski JH, Gitonga CW, Noor AM, Snow RW. The use of
schools for malaria surveillance and programme evaluation in Africa. Malar J. 2009;8:231. 29. Ministry of Education and Vocational Training. Number of primary
Schools by District Gov. Open Data Portal. 2015. http://opendata.go.tz/
dataset/idadi-ya-shule-za-msingi-kimkoa. Accessed 12 Dec 2015. 10. Stevenson JC, Stresman GH, Gitonga CW, Gillig J, Owaga C, Marube E,
et al. Reliability of school surveys in estimating geographic variation
in malaria transmission in the Western Kenyan Highlands. PLoS ONE. 2013;8:e77641. 30. Prime Minister’s Office, Regional Administration and Local Government
(Tanzania). Pre-Primary, Primary and Secondary Education Statistics 2013,
Regional Data. Dodoma. 2014. 11. Gitonga CW, Karanja PN, Kihara J, Mwanje M, Juma E, Snow RW, et al. Implementing school malaria surveys in Kenya: towards a national
surveillance system. Malar J. 2010;9:306. 31. Tanzania National Bureau of Statistics. National Bureau of Statistics. Tan-
zan. Natl. Bur. Stat. Stat. Dev. 2017. http://www.nbs.go.tz. Accessed 8 Jun
2017. y
12. Swana EK, Yav TI, Ngwej LM, Mupemba BN, Mukeng CK, et al. Author details
1 A language and environment for statistical computing. R
Foundation for Statistical Computing. Vienna, Austria; 2016. 42. Tanzania National Bureau of Statistics. National Bureau of Statistics (Tan-
zania). Statistics for Development. 2017. http://www.nbs.go.tz. Accessed
8 Jun 2017. 19. Sarpong N, Owusu-Dabo E, Kreuels B, Fobil JN, Segbaya S, Amoyaw F,
et al. Prevalence of malaria parasitaemia in school children from two dis-
tricts of Ghana earmarked for indoor residual spraying: a cross-sectional
study. Malar J. 2015;14:260. 43. WorldPop Project. Population density. Worldpop. 2015. http://www.world
pop.org.uk. Accessed 08 Jun 2017. 44. Gosoniu L, Msengwa A, Lengeler C, Vounatsou P. Spatially explicit burden
estimates of malaria in Tanzania: bayesian geostatistical modeling of the
malaria indicator survey data. PLoS ONE. 2012;7:e23966. y
20. Mathanga DP, Halliday KE, Jawati M, Verney A, Bauleni A, Sande J, et al. The high burden of malaria in primary school children in southern
Malawi. Am J Trop Med Hyg. 2015;93:779–89. 45. Hagenlocher M, Castro MC. Mapping malaria risk and vulnerability in the
United Republic of Tanzania: a spatial explicit model. Popul Health Metr. 2015;13:2. 21. Ndyomugyenyi R, Kroeger A. Using schoolchildren’s reports of bed net
use monitored by schoolteachers as a proxy of community coverage in
malaria endemic areas of Uganda. Trop Med Int Health. 2007;12:230–7. 46. Nankabirwa J, Brooker SJ, Clarke SE, Fernando D, Gitonga CW, Schel-
lenberg D, et al. Malaria in school-age children in Africa: an increasingly
important challenge. Trop Med Int Health. 2014;19:1294–309. 22. National Malaria Control Programme (Tanzania), Tanzania. National
Malaria Strategic Plan 2014–2020. Tanzania: Ministry of Health and Social
Welfare. 2014. 47. Pullan RL, Bukirwa H, Staedke SG, Snow RW, Brooker S. Plasmodium infec-
tion and its risk factors in eastern Uganda. Malar J. 2010;9:2. 23. Tanzania Commission for AIDS, Zanzibar AIDS Commission (Tanzania),
National Bureau of Statistics (Tanzania), Office of the Chief Government
Statistician (Tanzania), Macro International. Tanzania HIV/AIDS and Malaria
Indicator Survey 2007–2008. Dar es Salaam. 2008. http://dhsprogram
.com/pubs/pdf/AIS6/AIS6.pdf. Accessed 3 Jul 2017. 48. Carneiro I, Roca-Feltrer A, Griffin JT, Smith L, Tanner M, Schellenberg JA,
et al. Age-patterns of malaria vary with severity, transmission intensity
and seasonality in sub-Saharan Africa: a systematic review and pooled
analysis. PLoS ONE. 2010;5:e8988. 49. Gitonga CW, Edwards T, Karanja PN, Noor AM, Snow RW, Brooker SJ. Plasmodium infection, anaemia and mosquito net use among school
children across different settings in Kenya. Trop Med Int Health. 2012;17:858–70. 24. Author details
1 School-
based malaria prevalence: informative systematic surveillance measure
to assess epidemiological impact of malaria control interventions in the
Democratic Republic of the Congo. Malar J. 2018;17:141. 32. National Malaria Control Programme (Tanzania). National Guidelines for
Diagnosis and Treatment of Malaria. Malaria Control Series 25. 2014. 33. StataCorp. Stata Statistical Software: release 13. College Station: StataCorp
LP; 2013. 13. Ashton RA, Kefyalew T, Batisso E, Awano T, Kebede Z, Tesfaye G, et al. The
usefulness of school-based syndromic surveillance for detecting malaria
epidemics: experiences from a pilot project in Ethiopia. BMC Public
Health. 2016;16:20. 34. QGIS Development Team (2016). QGIS Geographic Information System. Open Source Geospatial Foundation Project. 35. U.S. Releases Enhanced Shuttle Land Elevation Data. https://www2.jpl. nasa.gov/srtm. Accessed 8 Jun 2017. 14. Okebe J, Affara M, Correa S, Muhammad AK, Nwakanma D, Drakeley
C, et al. School-based countrywide seroprevalence survey reveals
spatial heterogeneity in malaria transmission in the Gambia. PLoS ONE. 2014;9:e110926. 36. WorldClim, Global Climate Data. Free climate data for ecological model-
ling and GIS. http://www.worldclim.org. Accessed 13 Sep 2018. 37. Hijmans RJ, Cameron SE, Parra JL, Jones PG, Jarvis A. Very high-resolution
interpolated climate surfaces for global land areas. Int J Climatol. 2005;25:1965–78. 15. Nankabirwa J, Wandera B, Kiwanuka N, Staedke SG, Kamya MR, Brooker
SJ. Asymptomatic Plasmodium infection and cognition among primary
schoolchildren in a high malaria transmission setting in Uganda. Am J
Trop Med Hyg. 2013;88:1102–8. 38. Food and Agriculture Organization (FAO). Global ecological zoning for
the global forest resources assessment 2000. http://www.fao.org/docre
p/006/ad652e/ad652e10.htm. Accessed 19 Aug 2016. 16. Walldorf JA, Cohee LM, Coalson JE, Bauleni A, Nkanaunena K, Kapito-
Tembo A, et al. School-age children are a reservoir of malaria infection in
Malawi. PLoS ONE. 2015;10:e0134061. 39. Gething PW, Van Boeckel TP, Smith DL, Guerra CA, Patil AP, Snow RW,
et al. Modelling the global constraints of temperature on transmission of
Plasmodium falciparum and P. vivax. Parasit Vectors. 2011;4:92. 17. Takem EN, Affara M, Amambua-Ngwa A, Okebe J, Ceesay SJ, Jawara M,
et al. Detecting foci of malaria transmission with school surveys: a pilot
study in the Gambia. PLoS ONE. 2013;8:e67108. 40. Malaria Atlas Project. Malaria Atlas Project. 2016. http://www.map.ox.ac. uk. Accessed 18 Aug 2016. 18. Houngbedji CA, Chammartin F, Yapi RB, Hürlimann E, N’Dri PB, Silué KD,
et al. Spatial mapping and prediction of Plasmodium falciparum infec-
tion risk among school-aged children in Côte d’Ivoire. Parasit Vectors. 2016;9:494. 41. R Core Team. R. 58. Coughlin SS. Recall bias in epidemiologic studies. J Clin Epidemiol.
1990;43:87–91. 57. Bowling A. Mode of questionnaire administration can have serious effects
on data quality. J Public Health. 2005;27:281–91. Chacky et al. Malar J (2018) 17:452 Chacky et al. Malar J (2018) 17:452 Page 16 of 16 57. Bowling A. Mode of questionnaire administration can have serious effects
on data quality. J Public Health. 2005;27:281–91. 51. Ashton RA, Kefyalew T, Tesfaye G, Pullan RL, Yadeta D, Reithinger R, et al. School-based surveys of malaria in Oromia Regional State, Ethiopia: a
rapid survey method for malaria in low transmission settings. Malar J. 2011;10:25. 58. Coughlin SS. Recall bias in epidemiologic studies. J Clin Epidemiol. 1990;43:87–91. 52. Doolan DL, Dobaño C, Baird JK. Acquired immunity to malaria. Clin Micro-
biol Rev. 2009;22:13–36. 59. Kapesa A, Kweka EJ, Zhou G, Atieli HE, Kamugisha E, Mazigo HD, et al. Utility of passive malaria surveillance in hospitals as a surrogate to com-
munity infection transmission dynamics in western Kenya. Arch Public
Health. 2018;76:39. 53. Roll Back Malaria. Continuous LLIN Distribution—lessons in brief no. 9—
Tanzania. RBM. 60. Males S, Gaye O, Garcia A. Long-term asymptomatic carriage of Plas-
modium falciparum protects from malaria attacks: a prospective study
among Senegalese children. Clin Infect Dis. 2008;46:516–22. 54. Gitonga CW, Kihara JH, Njenga SM, Awuondo K, Noor AM, Snow RW, et al. Use of rapid diagnostic tests in malaria school surveys in Kenya: does
their under-performance matter for planning malaria control? Am J Trop
Med Hyg. 2012;87:1004–11. 61. Buchwald AG, Sorkin JD, Sixpence A, Chimenya M, Damson M, Wilson
ML, et al. Association between age and Plasmodium falciparum infection
dynamics. Am J Epidemiol. 2018. https://doi.org/10.1093/aje/kwy213
(Epub ahead of print). 55. Bousema T, Okell L, Felger I, Drakeley C. Asymptomatic malaria infections:
detectability, transmissibility and public health relevance. Nat Rev Micro-
biol. 2014;12:833–40. 56. Hay SI, Smith DL, Snow RW. Measuring malaria endemicity from intense
to interrupted transmission. Lancet Infect Dis. 2008;8:369–78. •
fast, convenient online submission
•
thorough peer review by experienced researchers in your field
•
rapid publication on acceptance
•
support for research data, including large and complex data types
•
gold Open Access which fosters wider collaboration and increased citations
maximum visibility for your research: over 100M website views per year
•
At BMC, research is always in progress. Learn more biomedcentral.com/submissions
Ready to submit your research ? Author details
1 Tanzania Commission for AIDS, Zanzibar AIDS Commission (Tanzania),
National Bureau of Statistics (Tanzania), Office of the Chief Government
Statistician (Tanzania), ICF International 2013. Tanzania HIV/AIDS and
Malaria Indicator Survey 2011–2012. Dar es Salaam. 2013. http://dhspr
ogram.com/pubs/pdf/AIS11/AIS11.pdf. Accessed 3 Jul 2017. 50. Kepha S, Nikolay B, Nuwaha F, Mwandawiro CS, Nankabirwa J, Ndibazza
J, et al. Plasmodium falciparum parasitaemia and clinical malaria among
school children living in a high transmission setting in western Kenya. Malar J. 2016;15:157. 25. Ministry of Health, Community Development, Gender, Elderly and
Children (Tanzania), Ministry of Health (Zanzibar), National Bureau of
Statistics (Tanzania), Office of Chief Government Statistician (Tanzania),
ICF International. Tanzania Demographic and Health Survey and Malaria Chacky et al. Malar J (2018) 17:452 Chacky et al. Malar J (2018) 17:452 Choose BMC and benefit from: •
fast, convenient online submission
•
thorough peer review by experienced researchers in your field
•
rapid publication on acceptance
•
support for research data, including large and complex data types
•
gold Open Access which fosters wider collaboration and increased citations
maximum visibility for your research: over 100M website views per year
•
At BMC, research is always in progress. Learn more biomedcentral.com/submissions
Ready to submit your research ? Choose BMC and benefit from:
|
https://openalex.org/W1971922163
|
https://www.scielo.br/j/ref/a/HxLDqqJfFKv9bs8RByXyZqv/?lang=pt&format=pdf
|
Portuguese
| null |
A instabilidade da díade XY: circuitando o dimorfismo sexual normativo
|
Revista Estudos Feministas
| 2,008
|
cc-by
| 2,797
|
260 Estudos Feministas, Florianópolis, 16(1): 147-163, janeiro-abril/2008 A instabilidade da díade XY
A instabilidade da díade XY
A instabilidade da díade XY
A instabilidade da díade XY
A instabilidade da díade XY: circuitando
: circuitando
: circuitando
: circuitando
: circuitando
o dimor
o dimor
o dimor
o dimor
o dimorfismo sexual normativo
fismo sexual normativo
fismo sexual normativo
fismo sexual normativo
fismo sexual normativo El descubrimiento de los
cromosomas sexuales. Un hito en
la historia de la Biología. condições ambientais e nutricionais em que o
embrião se desenvolveria), morfológica
(existiriam partículas nas células que
determinariam o sexo), e fisiológica (seria o
metabolismo celular no ovo o que produziria
um ou outro sexo). A autora mostra como o
descobrimento dos cromossomos sexuais em
1905 fez com que as teorias morfológicas se
impusessem sobre as demais, ao mesmo tempo
que o estudo da herança relacionada com os
cromossomos sexuais foi o ponto de partida da
genética cromossômica que Morgan e seus
colaboradores desenvolveram a partir de 1910,
considerada a segunda revolução genética, se
tomada a de Mendel como a primeira. DELGADO ECHEVERRÍA, Isabel. Como parte
do mesmo objetivo, propõe-se a determinar
onde e como foram realizados e quem realizou
esses descobrimentos e que meios os difundiram. Perseguindo argumentos críticos, polê-
micas e lutas entre diferentes campos do
conhecimento, a autora produz uma teia muito
bem articulada por uma rede de informações
colhidas de documentos históricos, da
bibliografia dos autores e das instituições, e
constrói assim uma produção de sentido que
dá clareza e lógica a uma nova leitura sobre as
teorias em suas várias dimensões, mas em
especial lhes é fiel, visibilizando-as para que
interajam com quem as lê. Seu segundo objetivo é colocar em
manifesto que os diversos conceitos, termos e
explicações da biologia atual que transmitem a
idéia da primazia masculina têm sua origem em
concepções defendidas em séculos anteriores,
assim como se esforça para dissociar o que foi
resultado do trabalho científico do que é a
projeção dos preconceitos sociais. Seu terceiro
objetivo de interesse para as gerações atuais e
futuras é o de constatar a existência de autoria
feminina nos descobrimentos biológicos do
passado. Nesse ponto, apresenta dados de
como a comunidade científica não foi justa com
as mulheres, quanto à concessão de prêmios e
honras ou de como o registro dos seus
descobrimentos ficou oculto, à sombra dos seus
maridos e colaboradores. A motivação para
este último objetivo advém do fato de que uma
das descrições de cromossomos sexuais
publicada em 1905 havia sido realizada por
mulheres. Assim, encontra-se no seu primeiro capítulo
a descrição e apresentação analítica da
tradição herdada até o século XIX, a partir do
mundo antigo e medieval, e os principais
pressupostos do mundo moderno. A autora
demonstra uma série de estudos com contra-
dições teóricas, experimentações ambíguas e
até discordantes demarcadoras do conheci-
mento biológico sobre os sexos. Demonstra
como padrões sociais influenciaram a produção
do conhecimento; como variantes na organiza-
ção sexual e no comportamento de machos e
fêmeas são tratadas como exceções a uma
norma que não se especifica; como as explica-
ções são prolíferas se a espécie em questão
altera formas de reprodução assexual com
indivíduos sexuados, e os contrapõem às
espécies em que machos são maiores do que
as fêmeas; e como as teorias silenciam quando
machos e fêmeas copulam e dão lugar a ovos
fecundados. DELGADO ECHEVERRÍA, Isabel. Naquela época, as
preocupações giravam em torno de diferentes
perspectivas, que são fundamentalmente três:
ambientalista (na qual o sexo dependeria das A posição dessa literatura crítica,
embora com a particularidade de estar nas
ciências chamadas naturais, se aproxima de
outras existentes no campo da determinação
sexual, como é o caso de Nelly Oudshoorn,1 em
suas análises sobre os contextos do desenvol-
vimento dos estudos sobre hormônios, o caso 260 Estudos Feministas, Florianópolis, 16(1): 147-163, janeiro-abril/2008 boa parte das formas de vida que se está
descrevendo. Para isso, faz um rastreamento
sistemático dos últimos anos da história da
biologia, origem das idéias atuais sobre a
determinação do sexo, bem como das chaves
de sua simplificação no processo de transmissão
e divulgação. Segue a trilha dos descobrimentos
dos cromossomos sexuais, para assinalar um
tempo e lugar e para estabelecer compara-
ções. Centra-se nas últimas décadas do século
XIX, considerando que os cromossomos sexuais
foram descritos pela primeira vez em 1905 e nas
primeiras décadas do século XX. de Cyntia Kraus,2 em seus estudos sobre a deter-
minação do sexo, o caso de Tamanini,3 em seu
capítulo de livro sobre a sexagem de embriões,
e de Ilana Lowy e Delphine Gardey,4 em seu
livro sobre a invenção do natural, entre outros
textos importantes no campo da ciência e
gênero. Seguindo o caminho do texto, verifica-se
como a autora apresenta um estudo particular
da história da biologia e do contexto do
descobrimento dos cromossomos sexuais, o que
permite perceber, mais uma vez, o que os
estudos de gênero têm insistentemente
demonstrado: que as diferenças entre os sexos
de homens e mulheres são estabelecidas não
por leis naturais biológicas, e que as diferenças
biológicas não são a origem das diferenças
sociais entre homens e mulheres e nem o
fundamento dos diferentes papéis que se
estabelecem para os indivíduos humanos de um
e de outro sexo. Ao longo de um texto denso e
enciclopédico, percorre o caminho sob a mira
de três objetivos: no primeiro, averigua o que
estimulou a curiosidade necessária para se
chegar a observar ao microscópio cromossomos
concretos, e como se chegou a constatar que
esses cromossomos eram diferentes entre
fêmeas e machos. Mesmo assim acontecendo,
investiga como conviveram em um mesmo
período histórico os descobrimentos sobre os
variados modos de reprodução e os diferentes
sistemas de determinação sexual. DELGADO ECHEVERRÍA, Isabel. Madrid: Consejo de Investigaciones
Científicas, 2007. 734 p. Madrid: Consejo de Investigaciones
Científicas, 2007. 734 p. Isabel Delgado Echeverría nasceu em
Zaragoza em 1960. É licenciada em Ciências
Biológicas pela Universidade Complutense de
Madrid. Trabalha desde 1985 como professora
de Biologia e Geologia no ensino secundário. Sua tese intitulada Las investigaciones sobre la
determinación del sexo y la diferencia sexual
(1891-1985) lhe deu o título de doutora em
Medicina, na Universidade de Zaragoza. Ela
pertence ao grupo Genciana (investigações em
gênero e ciência) do SIEM (Seminário Interdis-
ciplinar de Estudos da Mulher) da mesma
universidade. Em 1999 recebeu o prêmio de
investigação feminista “Concepción Gimeno de
Flaquer”, e em 2003 o prêmio “Dynamis de
Fomento de al Investigación en Historia de la
Medicina e de la Salud”. O tema do dimorfismo sexual normativo
constitui o eixo da preocupação central do livro,
embora existam outros focos que tornam esse
material fascinante. O seu apanhado histórico
seria já em si uma grande contribuição para os
estudos sobre a história da ciência, se
considerada a forma como a autora trabalha
com a riqueza documental e como elabora o
seu método de coleta, armazenamento,
organização e interpretação dos dados. Mas
sua riqueza se amplia muito, ao considerar-se a
original contribuição que ela oferece para os
estudos de gênero e ciência. A primeira razão a
demarcar é o fato de que a autora inclui nesse
seu trabalho a trajetória de várias mulheres
pesquisadoras, como Nettie Maria Stevens. Além
disso, é de extrema relevância e muito instrutivo
o modo como põe em destaque os traços do
androcentrismo, presente nos discursos da maior
parte dos trabalhos analisados. Seu livro El descubrimiento de los
cromosomas sexuales. Un hito en la historia
de la Biología, publicado pelo Consejo de
Investigaciones Científicas em Madrid, no ano
de 2007, com 734 páginas, analisa os trabalhos
científicos publicizados entre 1870 e 1940 em
relação aos problemas biológicos da
determinação do sexo. Estudos Feministas, Florianópolis, 16(1): 147-163, janeiro-abril/2008 261 DELGADO ECHEVERRÍA, Isabel. Para perseguir seus objetivos, a autora
apresenta, nos capítulos 1, 2 e 3, uma revisão
baseada na literatura histórica em que
contextualiza as idéias e teorias referentes à
determinação do sexo registradas na história da
ciência ocidental. Busca as raízes, conceitos e
preconceitos repassando a filosofia grega e A autora persegue ao longo dos capítulos
uma resposta à pergunta sobre como foram
construídos conceitos que se chocam com uma Estudos Feministas, Florianópolis, 16(1): 147-163, janeiro-abril/2008 261 dos de publicações anteriores. Segundo ela
mesma, a busca exaustiva inicial, que incluía os
séculos XIX e XX, lhe permitiu recompilar mais de
duas mil referências bibliográficas, cujo período
de publicação abarca de 1836 a 1985. detalha o século XIX, quando se produziram o
nascimento e o desenvolvimento da biologia
moderna. Analisa as conseqüências da teoria
mendeliana da hereditariedade em relação
com uma possibilidade de determinação
genética do sexo e confluência de distintas vias
de análise do problema a partir do ano de 1900. Sua análise centra-se nos artigos porque
estão mais próximos do processo de
descobrimento. Nesse aspecto, é possível
encontrar meticulosa análise bibliométrica,
sendo referida a quantidade, autores, países,
números e cidade das publicações. Há ainda
gráficos sobre a distribuição cronológica das
referências bibliográficas, o que permite localizar
como certos temas se concentram em
determinados períodos históricos, com a
predominância de certos idiomas de seus países
correspondentes. Essa metodologia bibliométri-
ca dá uma noção geral da literatura encontrada
e ensina sobre uma metodologia nem sempre
tão comum entre nós. Ao compor os desenhos,
não está apenas em consonância com outros
períodos históricos, mas também demonstra um
esforço investigativo revelador de temas novos
e do problema da determinação do sexo entre
as décadas de 1880 e 1930 que é original. p
p
No capítulo 4, detalha sua estratégia de
busca e de análise bibliográfica que a levaram
a recompilar 263 artigos científicos. Ela os
contrasta com a literatura histórica e os analisa
a partir das suas diferentes disposições teóricas
metodológicas, no que se refere às perspectivas
morfológica, ambientalista e fisiológica. Desenvolve esse capítulo, que denomina como
o contexto do descobrimento, a partir de investi-
gações e teorias sobre a fecundação, o
problema dos diferentes modos de reprodução
e a herança na determinação do sexo. DELGADO ECHEVERRÍA, Isabel. É um
capítulo apaixonante, com apresentação
detalhada dos autores do contexto das teorias
da fecundação, dos diferentes modos de
reprodução, da transmissão hereditária na
reprodução e da determinação do sexo. Localiza-se nele o desenvolvimento do
microscópio óptico e um enorme desenvolvi-
mento técnico que permitiu a descrição dos
gametas e gônadas da maior parte dos grupos
de organismos. Nos capítulos 6, 7 e 8, a autora se ocupa
da questão da partenogêneses a partir dos
conhecimentos
estabelecidos
pelas
investigações ocorridas entre 1878 e 1912 e que
colocavam em questão a necessidade da
atuação do espermatozóide. Pacientemente,
escrutina as diferentes polêmicas geradas pela
partenogêneses. Desenvolve a análise sobre a
perspectiva morfológica nessas discussões,
seguida das perspectivas ambientalista e
filosófica que são focadas nos capítulos 8 e 9. Isabel Delgado Echeverria trabalha de modo
organizado as muitas informações e a
magnitude da sua originalidade. Para as questões de gênero, é muito
interessante o modo como a autora constrói a
teia de sua rede e mostra o descobrimento de
óvulos e espermatozóides em todos os grupos
animais, vertebrados e invertebrados, e em seus
análogos, as plantas. Demonstra como a
descrição dos processos de fusão de gametas
na fecundação conduziu, ao final da década
de 1850, a um paradigma único sobre a
reprodução sexual no qual se unificavam os
reinos animal e vegetal, mesmo se, nas
considerações teóricas escritas por diferentes
investigadores, pudessem ser encontrados
pontos de vista mais ou menos mecanicistas,
morfológicos, fisiológicos e matizados com
diferentes graus de adesão ou crítica às teorias
da preformação ou epigêneses. Depreendem-se do seu texto várias
possibilidades analíticas, não apenas para
aumentar a valoração de cada aporte na
determinação do sexo, mas também para fazer
ver a situação desses aportes em relação aos
centros de interesse, às relações de força entre
instituições e países, assim como os principais
temas de debates, a evolução das idéias sobre
a determinação do sexo, o desenvolvimento
das ciências morfológicas e as considerações
ambientais. 262 Estudos Feministas, Florianópolis, 16(1): 147-163, janeiro-abril/2008 Referências bibliográficas
Referências bibliográficas
Referências bibliográficas
Referências bibliográficas
Referências bibliográficas fecundação. Esses estudos morfológicos,
especialmente o descobrimento dos cromos-
somos acessórios e sexuais, junto à hipótese
relativa ao dimorfismo dos gametas e sua
relação com a determinação do sexo, também
fizeram surgir entre os embriologistas a discussão
acerca do momento em que se produzia a
determinação ou fixação do caráter do sexo e
determinaram interesses confluentes entre
citologia e morfologia. A autora mostra como a
embriologia experimental é aplicada à questão
da determinação do sexo e como esses estudos
experimentais confluíram. fecundação. Esses estudos morfológicos,
especialmente o descobrimento dos cromos-
somos acessórios e sexuais, junto à hipótese
relativa ao dimorfismo dos gametas e sua
relação com a determinação do sexo, também
fizeram surgir entre os embriologistas a discussão
acerca do momento em que se produzia a
determinação ou fixação do caráter do sexo e
determinaram interesses confluentes entre
citologia e morfologia. A autora mostra como a
embriologia experimental é aplicada à questão
da determinação do sexo e como esses estudos
experimentais confluíram. KRAUS, Cynthia. “La bicatégorização par sexe à
l’epreuve de la science. Les cas des recher-
ches en biologie sur la détermination du sexe
chez les Humans”. In: LOWY, Ilana; GARDEY,
Delphine (Eds.). L’Invention du naturel, les
sciences et la fabrication du féminin et du
masculin. Paris: Éditions des Archives Contem-
poraines, 2000. p. 187-227. LOWY, Ilana; GARDEY, Delphine (Eds.). L’Invention
de naturel, les sciences et la fabrication
du féminin et du masculin. Paris: Editions des
Archives Contemporâneos, 2000. No capítulo 10, apresenta a participação
espanhola na biologia do sexo e demonstra que,
mesmo se considerados os conhecimentos
tardios nesse país, as contribuições espanholas
se sucederam rapidamente desde o primeiro
terço do século XX, ainda se consideradas as
conseqüências da sua interrupção de 1936 a
1939 por causa da guerra. Sua análise das
publicações espanholas, como o fez em relação
a outros países, permitiu à autora situar o
contexto institucional, as bibliografias dos autores,
a formação acadêmica dos profissionais, suas
relações com diferentes campos do conheci-
mento, bem como destacar as influências das
diferentes escolas de investigação. p
OUDSHOORN, Nelly. On the Making of Sex
Hormones: Research Materials and the
Production of Knowledge. Social Studies of
Science, London, Newbury Park and New
Delhi: Sage Periodical Press, v. 20, 1990. p. 5-
33. ______. DELGADO ECHEVERRÍA, Isabel. O texto também aborda as inúmeras
discussões estabelecidas entre os embriologistas
e fisiólogos que adotaram um ponto de vista
holístico e descartaram a influência dos fatores
ambientais com a adoção de uma perspectiva
“internalista” sobre a base de que o sexo estaria
já determinado no ovo antes e a partir da No capítulo 5, a autora ocupa-se do tema
da partogêneses, analisando documentos
originais a partir de variadas fontes de informação
primária e secundária: bibliografias de manuais
de história, biologia, medicina e ciência,
manuais universitários de diferentes disciplinas,
bibliografias de trabalhos monográficos, teses
de doutorado e licenciatura, catálogos de
diversas bibliotecas. Utiliza-se de redes de
informações a partir de documentos recupera- Referências bibliográficas
Referências bibliográficas
Referências bibliográficas
Referências bibliográficas
Referências bibliográficas “United We Stand: The Pharmaceutical
Industry, Laboratory, and Clinic in the
Development of Sex Hormones Into Scientific
Drugs, 1920-1940.” Science Techonology &
Human Values, London: Sage Periodical
Press, v. 18, n. 1, 1993. p. 5-23. ____. “Hormones, technique et corps: l’arche-
ologie des hormones sexuelles (1923-1940)”. Annales HSS, n. 4-5, p. 775-793, 1998. Para concluir, é preciso dizer que se trata
de um trabalho de riqueza incomensurável,
especialmente, no seu aspecto metodológico
e no modo como o material documental é
tratado, vindo a se constituir, nesse sentido, em
um bom exemplar de aprendizagem acadê-
mica para quem deseja trabalhar com fontes
semelhantes. Mas, além disso, é original e
fundamental para refletir sobre as teorias da
determinação do sexo e suas contribuições para
o campo dos estudos em gênero e ciência. ______. “Au sujet des corps, des technique et
des feminismes”. In: GARDEY, Delphine; LOWY,
Ilana (Eds.). L’invention du naturel, les scien-
ces et la fabrication du feminine et du mas-
culine. Paris: Éditions des Archives Contem-
poraines, 2000. p. 31- 44. OUDSHOORN, Nelly, and KIREJCZYK, Marta. Bodies
of Techonology: Women’s Involvement with
Reproductive Medicine. Georgia: Ohio State
University, 2000. p. 278-303. TAMANINI, Marlene. “¿Tecnología de óvulos y
espermatozoides
para
sexaje
de
embriones?” In: CARVALHO, Marília G. de;
TAMANINI, Marlene. Diversidad cultural,
género y tecnología: un abordage interdis-
ciplinario. Curitiba: Editora UTFPR, 2006. p. 121-158. Marlene Tamanini
Universidade Federal do Paraná Estudos Feministas, Florianópolis, 16(1): 147-163, janeiro-abril/2008 263 Notas
Notas
Notas
Notas
Notas 1 OUDSHOORN, 1990, 1993, 1998 e 2000; e Nelly
OUDSHOORN e Marta KIREJCZYK, 2000. 2 KRAUS, 2000. 3 TAMANINI, 2006. 4 LOWY e GARDEY, 2000. 1 OUDSHOORN, 1990, 1993, 1998 e 2000; e Nelly
OUDSHOORN e Marta KIREJCZYK, 2000. 2 KRAUS, 2000. 3 TAMANINI, 2006. 4 LOWY e GARDEY, 2000. Marlene Tamanini
Universidade Federal do Paraná Estudos Feministas, Florianópolis, 16(1): 147-163, janeiro-abril/2008 263
|
https://openalex.org/W4230060868
|
https://www.qeios.com/read/XIKEM3/pdf
|
English
| null |
Cutaneous Melanoma cM0 TNM Finding v8
|
Definitions
| 2,020
|
cc-by
| 61
|
Cutaneous Melanoma cM0 TNM Finding v8 National Cancer Institute Qeios ID: XIKEM3 · https://doi.org/10.32388/XIKEM3 Qeios · Definition, February 2, 2020 Open Peer Review on Qeios Source National Cancer Institute. Cutaneous Melanoma cM0 TNM Finding v8. NCI Thesaurus. Code C136895. Cutaneous melanoma without evidence of distant metastasis. LDH level is not applicable. (from AJCC 8th Ed.) Qeios ID: XIKEM3 · https://doi.org/10.32388/XIKEM3 1/1
|
https://openalex.org/W2947634950
|
https://europepmc.org/articles/pmc6589449?pdf=render
|
English
| null |
Low-dose aspirin as a promising agent for the prevention of spontaneous preterm birth
|
Evidence based nursing/Evidence-based nursing
| 2,019
|
cc-by
| 1,281
|
Commentary The hypothesis that aspirin might be effective for prevention of spon-
taneous preterm birth is based on overlapping pathophysiology with
preeclampsia. In this respect, two striking elements of the study by
Andrikopoulou et al should be addressed. First, the parent trial did not
show any benefit of low-dose aspirin with regards to preeclampsia.5 The
incidence of preeclampsia was 6.3% in the aspirin group and 4.6% in the
placebo group (RR 0.7; 95% CI 0.6 to 1.0). Second, the median gestational
age at randomisation was 20 weeks. Sensitivity analysis demonstrated
that gestational age at initiation of therapy did not influence the results
with regard to spontaneous preterm birth, while in case of preeclampsia
previous studies have shown that it is best to start with aspirin between
8 and 16 weeks of gestation.6 1Department of Obstetrics & Gynaecology, Amsterdam UMC, Vrije
Universiteit Amsterdam, Amsterdam Reproduction & Development,
de Boelelaan 1117, Amsterdam, the Netherlands, 2Department of
Obstetrics & Gynaecology, Amsterdam UMC, University of Amsterdam,
Amsterdam Reproduction & Development, Meibergdreef 9, Amsterdam,
the Netherlands Correspondence to: Dr. Martijn A Oudijk, Amsterdam UMC, University
of Amsterdam, Department of Obstetrics & Gynaecology, Amsterdam
Reproduction & Development, Meibergdreef 9, Amsterdam, the
Netherlands; m.a.oudijk@amc.uva.nl Andrikopoulou et al suggest that aspirin may either have a different or
additional mechanism of action in the prevention of spontaneous preterm
birth compared with its use in the prevention of preeclampsia. Initiation
of aspirin therapy between 8 and 16 weeks of gestation is important for
prevention of preeclampsia because this is the period that placentation
and transformation of spiral arteries occurs. We speculate that in case of
spontaneous preterm birth the anti-inflammatory properties of aspirin are
beneficial throughout pregnancy. Commentary on: Andrikopoulou M, Purisch SE, Handal-Orefice R,
et al. Low-dose aspirin is associated with reduced spontaneous
preterm birth in nulliparous women. Am J Obstet Gynecol. 2018;219:399.e1–399.e6. The results of studies performed thus far on the effect of aspirin for
prevention of spontaneous preterm birth are promising; however, more
placebo-controlled randomised controlled trials are needed to confirm these
results and to assess in which subgroups it is effective. To gain more insight
in the mechanism of action, we urge research groups to perform trans-
lational research alongside healthcare evaluation studies through biosam-
pling: that is, with blood samples and placenta or placental bed biopsies. Implications for practice and researchi ►
►
At this moment, there is insufficient evidence to implement low-dose
aspirin for the prevention of spontaneous preterm birth. ►
►
Placebo-controlled randomised controlled trials are needed to con-
firm the effectiveness of aspirin for the prevention of spontaneous
preterm birth. Correction notice This article has been corrected since it first published
online. The open access licence type has been amended. Methods The objective of the study by Andrikopoulou et al4 was to evaluate the
effect of low-dose aspirin for the prevention of spontaneous preterm
birth in nulliparous women without comorbidities. All were singleton
and gestations without congenital malformations. This was a secondary
analysis of a placebo-controlled randomised controlled trial preformed
from 1989 to 1991 with preeclampsia as primary outcome.5 Treatment
with aspirin or placebo was initiated between 13 and 25 weeks of gesta-
tion. The primary outcome of the present study was spontaneous preterm
birth <34 weeks of gestation. © Author(s) (or their employer(s)) 2019. Re-use permitted under CC BY. Published by BMJ. © Author(s) (or their employer(s)) 2019. Re-use permitted under CC BY. Published by BMJ. Open access This is an open access article distributed in accordance
with the Creative Commons Attribution 4.0 Unported (CC BY 4.0)
license, which permits others to copy, redistribute, remix, transform and
build upon this work for any purpose, provided the original work is
properly cited, a link to the licence is given, and indication of whether
changes were made. See: https://creativecommons.org/licenses/by/4.0/. Randomised controlled trial A total of 2543 women were included of which 1262 (49.6%) received
60 mg aspirin and 1281 (50.4%) placebo. Spontaneous preterm birth <34
weeks occurred significantly less in the aspirin-group (1.03%) compared
with the placebo-group (2.34%); RR 0.46 (95% CI 0.23 to 0.89). This effect
was independent of time of initiation of therapy (<16 vs ≥16 weeks) and
was also significant after exclusion of women who developed preeclampsia
(n=2). There was no difference in spontaneous preterm birth <37 weeks
of gestation. Low-dose aspirin as a promising
agent for the prevention of
spontaneous preterm birth Anadeijda J E M C Landman,1 Martijn A Oudijk2 Context Preterm birth is the most important cause of perinatal morbidity and
mortality worldwide. Despite the administration of progesterone to
women with a previous spontaneous preterm birth, ~20%–30% will
deliver preterm again.1 Accumulating evidence suggests that uteropla-
cental ischaemia plays an important role in the aetiology of spontaneous
preterm birth, comparable to its role in preeclampsia. As aspirin is proven
to reduce the risk of developing preeclampsia, it is hypothesised that
aspirin might also be effective to prevent spontaneous preterm birth
through its antithrombotic and anti-inflammatory effect. Several studies
have shown promising results.2–4 Competing interests None declared. Provenance and peer review Commissioned; internally peer reviewed. References 1. Jarde A, Lutsiv O, Beyene J, et al. Vaginal progesterone, oral progesterone, 17-OHPC,
cerclage, and pessary for preventing preterm birth in at-risk singleton pregnancies:
an updated systematic review and network meta-analysis. BJOG 2019;126. 82 Evid Based Nurs July 2019 | volume 22 | number 3 | 3. van Vliet EO, Askie LA, Mol BW, et al. Antiplatelet Agents and the Prevention of
Spontaneous Preterm Birth: A Systematic Review and Meta-analysis. Obstet Gynecol
2017;129:327–36. 2. Silver RM, Ahrens K, Wong LF, et al. Low-dose aspirin and preterm birth: a
randomized controlled trial. Obstet Gynecol 2015;125:876–84.
3. van Vliet EO, Askie LA, Mol BW, et al. Antiplatelet Agents and the Prevention of
Spontaneous Preterm Birth: A Systematic Review and Meta-analysis. Obstet Gynecol
2017;129:327–36.
4. Andrikopoulou M, Purisch SE, Handal-Orefice R, et al. Low-dose aspirin is
associated with reduced spontaneous preterm birth in nulliparous women. Am J
Obstet Gynecol 2018;219:399.e1–399.e6. 2. Silver RM, Ahrens K, Wong LF, et al. Low-dose aspirin and preterm bi
randomized controlled trial. Obstet Gynecol 2015;125:876–84. 4. Andrikopoulou M, Purisch SE, Handal-Orefice R, et al. Low-dose aspirin is
associated with reduced spontaneous preterm birth in nulliparous women. Am J Obstet Gynecol 2018;219:399.e1–399.e6. Evid Based Nurs July 2019 | volume 22 | number 3 | Commentary: Women's health and midwifery 2. Silver RM, Ahrens K, Wong LF, et al. Low-dose aspirin and preterm birth: a
randomized controlled trial. Obstet Gynecol 2015;125:876–84. 3. van Vliet EO, Askie LA, Mol BW, et al. Antiplatelet Agents and the Prevention of
Spontaneous Preterm Birth: A Systematic Review and Meta-analysis. Obstet Gynecol
2017;129:327–36. 4. Andrikopoulou M, Purisch SE, Handal-Orefice R, et al. Low-dose aspirin is
associated with reduced spontaneous preterm birth in nulliparous women. Am J
Obstet Gynecol 2018;219:399.e1–399.e6. 5. Sibai BM, Caritis SN, Thom E, et al. Prevention of preeclampsia with
low-dose aspirin in healthy, nulliparous pregnant women. The National
Institute of Child Health and Human Development Network
of Maternal-Fetal Medicine Units. N Engl J Med
1993;329:1213–8. 6. Bujold E, Roberge S, Lacasse Y, et al. Prevention of preeclampsia and intrauterine
growth restriction with aspirin started in early pregnancy: a meta-analysis. Obstet 2. Silver RM, Ahrens K, Wong LF, et al. Low-dose aspirin and pret
randomized controlled trial. Obstet Gynecol 2015;125:876–84. 3. van Vliet EO, Askie LA, Mol BW, et al. Antiplatelet Agents and the Prevention of
Spontaneous Preterm Birth: A Systematic Review and Meta-analysis. Obstet Gynecol
2017;129:327–36. 4. Andrikopoulou M, Purisch SE, Handal-Orefice R, et al. Low-dose aspirin is
associated with reduced spontaneous preterm birth in nulliparous women. Am J 83 Evid Based Nurs July 2019 | volume 22 | number 3 | Evid Based Nurs July 2019 | volume 22 | number 3 |
|
https://openalex.org/W3119165708
|
https://ijponline.biomedcentral.com/track/pdf/10.1186/s13052-020-00935-z
|
English
| null |
The child with medical complexity
|
The Italian Journal of Pediatrics/Italian journal of pediatrics
| 2,021
|
cc-by
| 5,865
|
© The Author(s). 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License,
which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give
appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if
changes were made. The images or other third party material in this article are included in the article's Creative Commons
licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons
licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain
permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the
data made available in this article, unless otherwise stated in a credit line to the data. REVIEW Open Access Abstract Children with medical complexity represent a big challenge for the physicians, their families and the society as well. Although there is no clear definition of this type of patients, they are affected by a chronic, often very severe
condition for their whole life. They also represent a huge cost for the health care system due to their needs of
continuous assistance. In this review we summarized the definitions of child with medical complexity. Then we illustrated the strategies to
treat and take care of these children in order to look at them not as a burden or a cost but as an opportunity to
growth and improve as clinicians and to improve the society, to give them the best life they can live. We also wanted to give voice to the physicians, the parents and the children themselves to really show and
understand what are their experiences and their feelings in dealing with their conditions. We concluded with the
description of one example of children with medical complexity: prematurity. We discussed the progresses in their
treatment so far in order to illustrate what is the future of pediatrics. Since it has been more and more
acknowledged that every child with medical complexity is unique, the future of pediatric is to organize an
individualized approach and to “see things with the eye of a child”. Keywords: Children, Pediatrics, Medical complexity, Individualized medicine, Chronic condition “Complex is a state of the world, complicated is a
state of the mind.”
Donald A. Norman present a univocal solution, but it needs to be globally
considered, analyzing all the elements that compose it
and their interactions. Complex and complicated: often in managerial lan-
guage these two concepts are used interchangeably. One
speaks about a “complex situation” or a “complicated
problem”, using the two concepts as synonyms. But they
are not. The child with medical complexity Manuela Gallo1, Rino Agostiniani2, Roberta Pintus1*
and Vassilios Fanos1,3 Manuela Gallo1, Rino Agostiniani2, Roberta Pintus1*
and Vassilios Fanos1,3 Gallo et al. Italian Journal of Pediatrics (2021) 47:1
https://doi.org/10.1186/s13052-020-00935-z Gallo et al. Italian Journal of Pediatrics (2021) 47:1
https://doi.org/10.1186/s13052-020-00935-z Introduction The term “complexity” derives from the Latin word
“complexus”, meaning something twisted, made by a
multiplicity of parts which are interdependent on one
another. A situation can be considered complex because
it originates from the intertwining of different elements
that interact with each other, creating disorder and
causing uncertainty. In a complex situation, it is difficult
to identify and manage all the variables involved, just as
much as it is essentially impossible to predict its
progresses. A problem that we define complex does not The term “complication” derives from the Latin “cum
plica” from which “complicare”, and it indicates some-
thing bent, wrapped around itself. A problem is compli-
cated when it shows itself as a result of a combination of
parts difficult to codify. Untangling the complication can
be exhausting, but there is a solution anyway. What is
complicated can be reduced to something simpler. We found it necessary to premise these definitions to
highlight the objective difficulty in managing the child
with medical complexity. In English literature they speak
about “complex chronic” or “medically complex” as well. * Correspondence: gomberta@icloud.com
1Università degli Studi di Cagliari, Cagliari, Italy
Full list of author information is available at the end of the article * Correspondence: gomberta@icloud.com
1Università degli Studi di Cagliari, Cagliari, Italy
Full list of author information is available at the end of the article Page 2 of 7 Page 2 of 7 Gallo et al. Italian Journal of Pediatrics (2021) 47:1 Gallo et al. Italian Journal of Pediatrics Gallo et al. Italian Journal of Pediatrics (2021) 47:1 detection of CMC, while the fourth cardinal point is rep-
resented by the “rare frequency” of this condition. We report as an example among the many cases of
child with medical complexity, that concerning a 10
years old child affected by CLOVES Syndrome (Congeni-
tal Lipomatous Overgrowth, Vascular malformations,
and Epidermal nevi): during his life he has undergone
16 major surgical procedures, 40 NMR, needed 14 differ-
ent types of Medical Specialist and he was forced to deal
with many travels with his family. More technically, the Children’s Hospital Associ-
ation, defines the CMC in groups of clinical risk
(CRG) of 5b degree or higher (as shown in Table 1),
including the following categories: Single Dominant
Chronic diseases, Complex Chronic diseases and Neo-
plastic diseases [5]. The most representative diagnoses include: chronic
lung diseases, genetic diseases, prematurity and its com-
plications,
congenital
cardiac
disturbances,
cerebral
palsy, neurological pathologies, orthopedic and respira-
tory diseases, HIV/AIDS, eating or metabolic disorders,
spina bifida, head trauma, sickle cell disease, short bowel
syndrome and congenital anatomical malformations. These diagnoses are often accompanied by one or more
comorbidities, such as developmental delay and intellec-
tual disability. The disability could be permanent or tem-
porary and affects without a doubt the life of the
children such that they cannot take part in the normal
activities that characterize the different phases of their
development. Successively, among all these examples, we
will
analyze
two
problems
among
those
above-
mentioned: prematurity and intellectual disability. Fur-
thermore, we will discuss the role of palliative care for
these children. How big the problem is? In the last years, the epidemiological landscape of the
pediatrics diseases has changed: we are facing a progres-
sive increase of the number of children suffering from
chronic diseases (children with special health care needs
[CSHCN]), mainly due to improvement in the level of
health cares and to the subsequent increase of the sur-
vival rate. Among the CSHCN, those with medical com-
plexity (CMC) are, without a doubt, the most fragile
from the medical point of view and with the most inten-
sive healthcare needs. The management of these patients
needs specific skills and new strategies that should in-
volve the pediatricians in the hospital as much as in the
homecare [1]. In the US, about 3 out of 76 millions of children shows
a medical complexity such that the category of CMC is
increasing at a rate of about 5% per year, exceeding the
growth rate of the children as a whole [1]. The daily assistance of the CMC is, thus, very problem-
atic and it develops on different levels, among which there
are 3 that are fundamental: drugs administration, respira-
tory and nutritional support. The security risks addressed
by the CMC are, for the most part, due to the complex na-
ture of their care rather than the specific risks posed by
the complicated nature of their pathologies. In particular, it has gone from 3.000.000 cases in 2013
to over 3.600.000 in 2017. In Italy, for example, the percentages of hospitalization
in 2012 at the Meyer Pediatric Hospital in Florence, are
represented for the 35.8% by children with congenital
malformation, and for the 22.7% by children with neuro-
developmental disorders. It must be strongly emphasized that, precisely by
virtue of these characteristics, these children highlight
the vulnerabilities of our approach to the cures: frag-
mented and often inefficient [6]. Who are the children with medical complexity? The Maternal and Child Health Bureau (MCHB) defined
the CMC those who are affected by or are at risk of de-
veloping a pathology that can alters the neurological de-
velopment, chronic behavioral or emotional problems
and who need diversified and more intense health care,
compared to that needed by the general pediatric popu-
lation [2]. Furthermore, although CMC represent < 1% of the
pediatric population, they use almost a third of all
pediatric expenses for the health care [7]. Data from US show that the most important items
of healthcare expenditure due to CMC are repre-
sented by the neurologic/neurosurgical diseases for
the 30%, by hematologic pathologic conditions for the
13%, by neonatal intensive care for more than the
10% but, even more interesting data, is that 1 out 4
hospitalization
of
these
little
patients
could
(and
should) be avoided [8]. There is still no clear definition of CMC, nevertheless,
to detect this kind of patients, in the English literature,
the following features are acknowledged and increasingly
accepted [3, 4]: 1) chronic and severe health conditions; Thus, a clear and shared strategy is necessary for the
management of CMC, the aims of which should be: im-
prove the experience of these children in the healthcare
facilities and the quality of the cares, improve the results
(for the children themselves and for their families) and
reduce the future healthcare costs [9]. 2) substantial need of the National Health Care
System; 3) functional limitation, often severe; 4) high utilization of health resources. It should be noted that in Italy, the first three above-
mentioned characteristics are acknowledged for the It should be noted that in Italy, the first three above-
mentioned characteristics are acknowledged for the Page 3 of 7 Gallo et al. Italian Journal of Pediatrics Gallo et al. Who are the children with medical complexity? Italian Journal of Pediatrics (2021) 47:1 Table 1 Classification of the Children’s Hospital Association Table 1 Classification of the Children’s Hospital Association
NON-CHRONIC
(CRG 1,2)
EPISODIC
CHRONIC
(CRG 3,4)
LIFELONG CHRONIC
(CRG 5a,5b)
COMPLEX CHRONIC
(CRG 6,7,9)
MALIGNANCIES
(CRG 8)
Description - Acute
pathological
condition
lasting less
than one year
- Pathological condition
lasting more than one year
that manifests itself in an
episodic manner;
- potentially treatable with
adequate treatment;
- likely not to manifest in
adulthood
- Severe primary pathological
condition, involving an entire
apparatus and more likely to
cause long-term damage
- Severe chronic
pathological
condition involving
two or more
apparatuses or
- chronic pathological
condition with
progressive
deterioration or
- technology-
dependent patho-
logical condition
- Highly malignancy
pathological
condition that
requires timely
treatment
Examples
- Head trauma,
peritonitis, UTI
- Asthma, major depression
- Congenital heart disease, short
bowel syndrome
-Cerebral palsy,
intellectual disability
- Neuroblastoma,
acute myeloid
leukaemia
Taken from “Children and Young Adults with Medical Complexity: Serving an Emerging Population”, Edwin F. Simpser et al. Modified [5]. COMPLEX CHRONIC
(CRG 6,7,9)
MALIGNANCIES
(CRG 8) In this sense, the “NACHRI”, (National Association of
Children’s Hospitals and Related Institutions) recently
elaborated a program that develops on three levels: A recent study showed, interestingly, the significant
change of the health care professionals after a training
course, with regard to the problems that we are illustrat-
ing (see Table 2) [11]. 1. Create a national database to measure the
assistance of CMC, relative costs and share
information The parents A scientific paper of 2019, revealed that the parents of
these little patients have a sense of great awareness of
the difficulties, from the health care professionals, to
take care of their children, but of the importance of their
role as well [12]. In this regard, we report three brief ob-
servations made by as many parents. 2. Create a regional network to share clinical data
with the different health care professionals who
assist these little patients p
3. Create domestic care services to mitigate the
severity of child’s chronic conditions “They are the professionals, we are the professionals of
our children.” Thus, the beginning of the care of CMC must be
precocious, global and extended to the family, the
management
must
be
multidisciplinary
(from
the
interaction to the integration) with a multi-specialized
staff and a pediatric coordination center and the net-
work of care must include the birth centers, the cen-
ters of references, the family pediatrician and the
territorial sanitary garrisons [10]. Many parents perceive the scarce collaboration from
the members of the staff and are suffering for the lack of
communication and interaction among the different
medical specialists. “A doctor often does not ask another doctor.” Furthermore, the parents are open to “negotiate” the
boundaries and the limits of their assistance. “When a nurse, see that you (parent), do the right
thing at the right moment, they are more inclined to
let you do (it).” The experience of the children with medical complexity,
of their parents, of the health care professionals, of the
school and the society Table 2 Characteristics that identify the perception of the child
with medical complexity, before and after a training course
Before the training
After the training
Chronic disease
Chronic condition
Multiple problems and drugs
Need for support for daily activity
Home technology
Technology addition
Follow up
Frequent appointments and
hospitalizations
Multidisciplinary approach
Team approach
Taken from [11]. Modified Table 2 Characteristics that identify the perception of the child
with medical complexity, before and after a training course
Before the training
After the training
Chronic disease
Chronic condition
Multiple problems and drugs
Need for support for daily activity
Home technology
Technology addition
Follow up
Frequent appointments and
hospitalizations
Multidisciplinary approach
Team approach
Taken from [11]. Modified The school The school plays a fundamental role in the difficult
growth path of the CMC and in supporting their
families. The health care professionals Often, the reactions of health care professionals to the
CMC tend to be elusive: “I do not have time to deal with
all this”, “these pathologies are unfamiliar to me”, “I am
not paid to do all this”. Therefore, the CMC switches
from the management by the pediatrician from that by
the specialist. It is very important that the health care
professionals think about their behaviors towards the pa-
tients, in particular those with chronic diseases, and even
more, the little patients with medical complexity. Page 4 of 7 Page 4 of 7 Gallo et al. Italian Journal of Pediatrics (2021) 47:1 Gallo et al. Italian Journal of Pediatrics (2021) 47:1 Gallo et al. Italian Journal of Pediatrics admissions and provides an easier access to care beyond
the state borders, when needed. Thus, the collaboration between the health care pro-
fessionals and the parents is essential, in particular in
case of CMC, whose parents are specialized health care
professionals themselves. In addition to the reinforced
partnerships with the health care professionals, the
needs expressed by the parents include a better commu-
nication with the staff and a greater attention to the con-
tinuity of care. In fact, the participation of the families in
the cures and the assistance of their own child, repre-
sents one of the most significant indicators of quality of
the Healthcare System. One example of children with medical complexity
The preterm newborn In this respect, the CAPHC (Canadian Association of
Paediatric Health Centers), has done an excellent job in
April 2018, creating some Guidelines concerning the
management of these little patients, including the school
environment as well [13]. Let us consider the prematurity and its complications. In
the last 10 years the percentage of prematurity has
remained virtually unchanged (1 out of 10 new births),
while the percentage of very preterm newborns that are
part of CMC, has increased. This increment is strictly
related to, on one hand to the increase of the artificial
fertilization techniques, and on the other hand, to the
improvement of the quality of the care that allows for a
reduction of the perinatal mortality even in case of very
low gestational age and greater outcomes in the long
term as well. In Italy, the scholastic integration of the students
with physical and intellectual disability constitutes a
strength point of our educational system, aiming at
the creation of a welcoming community in which all
students, despite their functional diversity, can experi-
ence individual and social growth. Full inclusion of
the students with disabilities is a goal that the school
pursues through an intense and articulate planning,
valorizing the internal professionalisms and the re-
sources offered by the territory. The MIUR imple-
ments
several
support
measures
to
favor
the
integration: support teachers, funding of projects and
activities for the integration, training initiatives for
support curricular teachers and the administrative,
technical and auxiliary staff [14]. There
is
a
very
spirited
debate
concerning
the
“dawn of life” in the literature: Do we really want
what we want? [16]. It is very difficult to objectively define the limit of the
ability to survive: scientific criteria are formulated in
very imprecise terms and they depend on the available
technological resources. However, it should be stressed that, even in more de-
veloped countries, the care of CMC is too often “react-
ive” rather than “preventive” or “anticipatory”, therefore
the path for an effective and holistic assistance is to date
still very long. In Japan, for instance, they are investing many re-
sources to improve the survival rate of the newborns
born between the 22 and 23 weeks of gestation, although
the knowledge and the medical technologies are limited
for the care of these neonates and on the long-term
outcomes [17]. The patients It is in no way possible to describe in an objective and
timely manner the life of a CMC, their emotions and
their ambitions. Let us therefore leave room for the
words of a patient, an American young adolescent, who
admirably synthesize everything you need to know: “Since the team that takes care of me, taught me the
abilities that I needed, I am independent and I will be
able to go on with my adult life with the necessary sup-
ports. I will soon be an adult with medical complexity
and the way to self-manage, that I learned, will allow me
to satisfy my needs and reach my goals” L. Rodgers. Society and politics Several studies have suggested that the therapies for
the treatment of the preterm infant may have conse-
quences in adult life. For instance, it is known that until
the early 1990s, the majority of preterm newborns, were
extensively treated with corticosteroids to prevent or re-
duce the incidence and severity of chronic lung disease
[18]. In fact, corticosteroids improve respiratory function
and reduce the need for both mechanical ventilation and
oxygen administration, resulting in a decreased mortal-
ity. However,
recent
studies
have
shown
that
the
treatment with corticosteroids may also have negative
long-term effects on the heart, possibly initiated by A detailed examination of this aspect goes beyond our
objectives. Nevertheless, it is interesting to note that some States
have made considerable progresses in attempting to
guarantee an adequate assistance to CMC. For example,
the American society has taken a huge step forward in
2019 by making law the ACE Kids Act (Advancing Care
for Exceptional Kids) [15]. The ACE Kids Act supports a better coordination of
the CMC cares, the home assistance and in the different
places of the community, reduces the unnecessary Gallo et al. Italian Journal of Pediatrics (2021) 47:1 Page 5 of 7 Gallo et al. Italian Journal of Pediatrics but where premature death is still possible (for
example cystic fibrosis, HIV/AIDS, cardiovascular
anomalies, extreme prematurity); but where premature death is still possible (for
example cystic fibrosis, HIV/AIDS, cardiovascular
anomalies, extreme prematurity); inhibiting the cardiomyocyte mitosis, and resulting in a
reduced number of cardiac muscle cells in adulthood. These changes appear to elicit a late systolic dysfunction,
due to compensatory cardiac dilation [19]. category 3: children with progressive conditions
without curative options, where treatment is
exclusively palliative and may commonly extend
over many years (for example neuromuscular or
neurodegenerative disorders, progressive metabolic
disorders, chromosomal abnormalities, advanced
metastatic cancer on first presentation); Moreover, preterm infants, given their low nephrons
number, are considered at a higher risk of congenital oli-
gonephronia, as well as low birth weight neonates, link-
ing body weight at birth with the susceptibility to
develop CKD in adulthood [20]. Many studies have shown that prematurity is a risk fac-
tor for the development of neurological diseases, such as
Parkinson’s and Alzheimer’s disease in later life [21]. Pediatric palliative care In Italy the numbers of patients involved in hospital
stays for LLC and life-threatening diseases eligible for
PPC were 35,734 in 2001, 37,040 in 2002 and 38,641 in
2003. For the patients with non-oncological diseases, the
maximum rate occurred in children aged 0 to 2 years,
with a peak in the first month of life [27]. Life-limiting conditions (LLCs) in children have been de-
fined as conditions for which there is no reasonable hope
of cure and from which children will die. Life-threatening
conditions are those for which curative treatment may be
feasible but can fail, such as cancer [23]. Palliative care for children and young people (PPC)
with life-limiting or life-threatening conditions is an ac-
tive and total approach to care, from the point of diag-
nosis throughout the child’s life and death. It embraces
physical, emotional, social, and spiritual elements, and
focuses on enhancing quality of life for the child/young
person and supporting their family [24]. The need for PPC globally is unknown. Making accur-
ate estimates is important to understand the scope of
the need and to advocate to meet it [28]. These children can be considered part of CMC. The main elements of children’s palliative care services
include: medical care, 24 h nursing care, social care, edu-
cation, play, therapies, short breaks, leisure and emo-
tional support. There needs to be co-ordination of these
services to enable seamless and holistic support for
families. According to the Association for Children’s Palliative
Care (ACT) and the Royal College of Paediatrics and
Child Health (RCPCH), we can identify four categories
of children eligible for PPC: Global institutions and interested people in children’s
health and well-being should be more interested in and
make PPC accessible to all children who need it. category 1: children with life-threatening conditions
for which curative treatment may be possible, but
can fail (for example cancer, organ failure of heart,
liver or kidney, infections); y
category 2: children with conditions requiring long
periods of intensive treatment aimed atprolonging
life and allowing participation in normal activities, Society and politics category 4: children with irreversible, not
progressive conditions with severe disability causing
extreme vulnerability to health complications (for
example severe cerebral palsy, genetic disorders,
congenital malformations, prematurity, brain or
spinal cord injury) [25]. Cortical development is a complex process that follows
a well-defined chronology: neurogenesis and neuronal mi-
gration continue until the end of the second trimester
while synaptogenesis, brain folding, and myelination are in
progress after premature birth. Consequently, preterm
newborns are predisposed to disruptions in cortical con-
nectivity, cell death, and myelination disorders. The nega-
tive effects of these processes may clinically appear only
after a long time, during adolescence or in adult life [22]. Medical and technological progress allow newborns,
children and teenagers suffering from lifelimiting and
life-threatening illnesses to survive, but not necessarily
to recover. Therefore, in recent years, the number of
children needing PPC has been steadily increasing. These and many other examples make us understand
that we need to pay close attention to CMCs. Diseases
that manifest during the early stages of life, such as hap-
pens in preterm babies, can also have important conse-
quences on their future as adults. In fact, complications
of prematurity may influence the quality of life, with ser-
ious long-term consequences in adulthood. Several scientific studies have shown this. A British study estimates that more than 40,000 chil-
dren (0–19 years) in England in 2009/2010 were living
with an LLC [23]. A more recent study shows that these
children are more than 86,000 in 2017/2018 [26]. The
highest requirement of children with LLCs for palliative
care occurs in the first year of life and decreases during
childhood [23, 26]. Conclusions In this review we have outlined some fixed points in the
definition, in the approach and in the experience of Page 6 of 7 Gallo et al. Italian Journal of Pediatrics (2021) 47:1 Page 6 of 7 Gallo et al. Italian Journal of Pediatrics CMC and in the implications of this complexity for fam-
ilies, health professionals and society. 3. Srivastava R, Stone BL, Murphy NA. Hospitalist care of the medically
complex child. Pediatric Clin North Am. 2005;52(4):1165–87. 3. Srivastava R, Stone BL, Murphy NA. Hospitalist care of the medically
complex child. Pediatric Clin North Am. 2005;52(4):1165–87. 4. Cohen E, Kuo DZ, Agrawal R, Berry JG, Bahgat SKM, Simon TD, Srivastava R. Children with medical complexity: an emerging population for clinical and
research initiatives. Pediatrics. 2011;127:529–38. 4. Cohen E, Kuo DZ, Agrawal R, Berry JG, Bahgat SKM, Simon TD, Srivastava R. Children with medical complexity: an emerging population for clinical and
research initiatives. Pediatrics. 2011;127:529–38. The Pediatrics of the future is more and more techno-
logical and advanced but “in medicine, the high tech,
meaning the super-technology, cannot and must not
make us forget the high touch, meaning the human con-
nection” (J. Naisbitt, High Tech/High Touch) [29]. 5. Simpser E, Budo P, PCCA association. Children and Young Adults with
Medical Complexity: Serving an Emerging Population 2016. https://
pediatriccomplexcare.org. 5. Simpser E, Budo P, PCCA association. Children and Young Adults with
Medical Complexity: Serving an Emerging Population 2016. https://
pediatriccomplexcare.org. 6. Schindlel C, Pordes E, Finkenbinder S, Lee K. Safety in children with medical
complexity: our canaries in the coal mine? Curr Treat Options Pediatrics. 2019;5(2):165–82. The goal of the Pediatrician is to take care of the chil-
dren, but also to communicate with them and the family
and establish a therapeutic alliance and, not least, “see
things with the eyes of the child” [30]. 7. Cohen E, Berry JG, Camacho X, Anderson G, Wodchis W, Guttmann A. Patterns and costs of health care use of children with medical complexity. Pediatrics. 2012;130(6):e1463–70. 7. Cohen E, Berry JG, Camacho X, Anderson G, Wodchis W, Guttmann A. Patterns and costs of health care use of children with medical complexity. Pediatrics. 2012;130(6):e1463–70. 8. Berry JG, Hall M, Neff J, Goodman D, Cohen E, Agrawal R, Kuo D, Feudtner
C. Children with medical complexity and Medicaid: spending and cost
savings. Health affairs (Project Hope). 33(12):2199–206. Authors’ contributions 18. Bassareo PP, Abella R, Fanos V, Mercuro G. Biomarkers of corticosteroid-
induced hypertrophic cardiomyopathy in preterm infants: implications for
pathophysiology and prognosis. Front Biosci (Elite Ed). 2010;2:1460–71. Manuela Gallo helped drafting the manuscript. Rino Agostiniani substantially
revised the paper. Roberta Pintus revised the paper and checked the
references. Vassilios Fanos had the original idea and helped drafting the
work and substantially revised the paper. All authors read and approved the
manuscript. 19. Bassareo PP, Namana V, Fanos V, Mercuro G. Preterm birth and risk of heart
failure up to early adulthood. J Am Coll Cardiol. 2017;70(15):1943–4. 19. Bassareo PP, Namana V, Fanos V, Mercuro G. Preterm birth and risk of heart
failure up to early adulthood. J Am Coll Cardiol. 2017;70(15):1943–4. 20. Fanos V, Castagnola M, Faa G. Prolonging nephrogenesis in preterm infants:
a new approach for prevention of kidney disease in adulthood? Iran J
Kidney Dis. 2015 May;9(3):180–5. 20. Fanos V, Castagnola M, Faa G. Prolonging nephrogenesis in preterm infants:
a new approach for prevention of kidney disease in adulthood? Iran J
Kidney Dis. 2015 May;9(3):180–5. (Sanctity
of
life
or
quality
of
life?
Singer,
Pediatrics 1983) (Sanctity
of
life
or
quality
of
life? Singer,
Pediatrics 1983) 12. Rennick JE, St-Sauveur I, Knox AM, Ruddy M. Exploring the experiences of
parent caregivers of children with chronic medical complexity during
pediatric intensive care unit hospitalization: an interpretive descriptive
study. BMC Pediatr. 2019;19(1):272. 12. Rennick JE, St-Sauveur I, Knox AM, Ruddy M. Exploring the experiences of
parent caregivers of children with chronic medical complexity during
pediatric intensive care unit hospitalization: an interpretive descriptive
study. BMC Pediatr. 2019;19(1):272. Author details
1 1Università degli Studi di Cagliari, Cagliari, Italy. 2Area Pediatria e
Neonatologia della Asl Toscana Centro (Firenze), Florence, Italy. 3Terapia
Intensiva Neonatale, AOU e Università degli Studi di Cagliari, Cagliari, Italy. 1Università degli Studi di Cagliari, Cagliari, Italy. 2Area Pediatria e
Neonatologia della Asl Toscana Centro (Firenze), Florence, Italy. 3Terapia
Intensiva Neonatale, AOU e Università degli Studi di Cagliari, Cagliari, Italy. 24. Together for Short Lives. A Guide to Children’s Palliative Care. Bristol, 2018. Available at: https://www.togetherforshortlives.org.uk/wp-content/uploads/2
018/03/TfSL-A-Guide-to-Children%E2%80%99s-Palliative-Care-Fourth-Edition-
FINAL-SINGLE-PAGES.pdf. Received: 28 May 2020 Accepted: 16 November 2020 25. ACT. A guide to the development of children’s palliative care services. Association for Children with Life-Threatening or Terminal Conditions and
their Families (ACT) and the Royal College of Paediatrics and Child Health
(RCPCH). 3rd ed; 2009. Competing interests The authors declare that they have no competing interest. The authors declare that they have no competing interest. 23. Fraser LK, Miller M, Hain R, Norman P, Aldridge J, McKinney PA, Parslow RC. Rising national prevalence of life-limiting conditions in children in England. Pediatrics. 2012;129(4):e923–9. 23. Fraser LK, Miller M, Hain R, Norman P, Aldridge J, McKinney PA, Parslow RC. Rising national prevalence of life-limiting conditions in children in England. Pediatrics. 2012;129(4):e923–9. Availability of data and materials 21. Scafidi J, Fagel DM, Ment LR, Vaccarino FM. Modeling premature brain injury
and recovery. Int J Dev Neurosci. 2009 Dec.;27:863–71. 21. Scafidi J, Fagel DM, Ment LR, Vaccarino FM. Modeling premature brain injury
and recovery. Int J Dev Neurosci. 2009 Dec.;27:863–71. “Data sharing is not applicable to this article as no datasets were generated
or analysed during the current study. 22. Faa G, Manchia M, Pintus R, Gerosa C, Marcialis MA, Fanos V. Fetal
programming of neuropsychiatric disorders. Birth Defects Res C Embryo
Today. 2016 Sep;108(3):207–23. 22. Faa G, Manchia M, Pintus R, Gerosa C, Marcialis MA, Fanos V. Fetal
programming of neuropsychiatric disorders. Birth Defects Res C Embryo
Today. 2016 Sep;108(3):207–23. Abbreviations 13. Canadian Association of Paediatric Health Centres (CAPHC), Complex Care
Community of Practice. CAPHC Guideline for The Management of Medically
Complex Children & Youth Through the Continuum of Care. 2019. 13. Canadian Association of Paediatric Health Centres (CAPHC), Complex Care
Community of Practice. CAPHC Guideline for The Management of Medically
Complex Children & Youth Through the Continuum of Care. 2019. CLOVES: Congenital Lipomatous Overgrowth Vascular malformation and
Epidermal nevi; CSHCN: Children with Special Health Care Needs;
CMC: Children with Medical Complexity; MCHB: Maternal and Child Health
Bureau; CRG: Clinical Risk Group; NACHRI: National Association of Children’s
Hospital and Related Institution; CAPHC: Canadian Association of Pediatric
Health Centers; MIUR: Ministero dell’Università e della Ricerca meaning
Ministry of education, university and research; ACE: Advancing Care for
Expectional kids; LLCs: Life-limiting conditions; PPC: Pediatric Palliative care;
ACT: Association for Children’s Palliative Care; RCPCH: Royal College of
Paediatrics and Child Health 14. MIUR. Linee guida per l'integrazione scolastica degli alunni con disabilità. 2009. Available at: http://www.edscuola.it/archivio/norme/circolari/nota_4_
agosto_09.pdf. Accessed 30 Mar 2020. 14. MIUR. Linee guida per l'integrazione scolastica degli alunni con disabilità. 2009. Available at: http://www.edscuola.it/archivio/norme/circolari/nota_4_
agosto_09.pdf. Accessed 30 Mar 2020. 15. ACE Kids Act: Policy Win of The Decade. Children’s Hospital Association. Available at: https://childhospassoc.maps.arcgis.com/apps/MapJournal/index. html?appid=0e85ef89f3634c46a0e924e41d9281bf. Accessed 07 Oct 2020. 15. ACE Kids Act: Policy Win of The Decade. Children’s Hospital Association. Available at: https://childhospassoc.maps.arcgis.com/apps/MapJournal/index. html?appid=0e85ef89f3634c46a0e924e41d9281bf. Accessed 07 Oct 2020. Expectional kids; LLCs: Life-limiting conditions; PPC: Pediatric Palliative care;
ACT: Association for Children’s Palliative Care; RCPCH: Royal College of
Paediatrics and Child Health 16. Krick JA, Feltman DM. Neonatologists' preferences regarding guidelines for
periviable deliveries: do we really know what we want? J Perinatol. 2019;
39(3):445–52. 16. Krick JA, Feltman DM. Neonatologists' preferences regarding guidelines for
periviable deliveries: do we really know what we want? J Perinatol. 2019;
39(3):445–52. Acknowledgments
not applicable 17. Nishida H. Improved Outcome of Infants Born at less than 24 Weeks of
Gestation in Japan. J Health Med Informat. 2013;S11:001. 17. Nishida H. Improved Outcome of Infants Born at less than 24 Weeks of
Gestation in Japan. J Health Med Informat. 2013;S11:001. Conclusions This is all the more true the more complex the
pathology. We cannot fail to mention, in conclusion, a famous
phrase by Singer: 9. Berry JG, Hall M, Cohen E, O'Neill M, Feudtner C. Ways to identify
children with medical complexity and the importance of why. J Pediatr. 2015;167(2):229–37. 9. Berry JG, Hall M, Cohen E, O'Neill M, Feudtner C. Ways to identify
children with medical complexity and the importance of why. J Pediatr. 2015;167(2):229–37. 10. Corsello G. Complessità assistenziale, documento congiunto SIP-AOPI. https://
www.sip.it/wp-content/uploads/2017/08/Documento-AOPI-SIP.pdf. 2017. 10. Corsello G. Complessità assistenziale, documento congiunto SIP-AOPI. https://
www.sip.it/wp-content/uploads/2017/08/Documento-AOPI-SIP.pdf. 2017. “Every human being has the right to the highest
quality of life he can achieve”. “Every human being has the right to the highest
quality of life he can achieve”. “Every human being has the right to the highest
quality of life he can achieve”. 11. Huth K, Long-Gagne S, Mader J, Sbrocchi A. Understanding the needs of
children with medical complexity. MedEdPORTAL. 2018;14:10709. 11. Huth K, Long-Gagne S, Mader J, Sbrocchi A. Understanding the needs of
children with medical complexity. MedEdPORTAL. 2018;14:10709. References 1. Agostiniani R, Nanni L, Langiano T. Children with medical complexity: the
change in the pediatric epidemiology. J Pediatr Neonat Individual Med. 2014;3(2):e030230. 26. Fraser L, Gibson-Smith D, Jarvis S, Norman P, Parslow R. 'Make every child
count'. Estimating current and future prevalence of children and young
people with life-limiting conditions in the United Kingdom. February 2020. Published Report:https://www.york.ac.uk/media/healthsciences/documents/
research/public-health/mhrc/Prevalence%20reportFinal.pdf. 26. Fraser L, Gibson-Smith D, Jarvis S, Norman P, Parslow R. 'Make every child
count'. Estimating current and future prevalence of children and young
people with life-limiting conditions in the United Kingdom. February 2020. Published Report:https://www.york.ac.uk/media/healthsciences/documents/
research/public-health/mhrc/Prevalence%20reportFinal.pdf. 2. McPherson M, Arango P, Fox H, Lauver C, McManus M, Newacheck PW,
et al. A new definition of children with special health care needs. Pediatrics. 1998;102(1 pt 1):137–40. 2. McPherson M, Arango P, Fox H, Lauver C, McManus M, Newacheck PW,
et al. A new definition of children with special health care needs. Pediatrics. 1998;102(1 pt 1):137–40. Page 7 of 7 Gallo et al. Italian Journal of Pediatrics (2021) 47:1 Gallo et al. Italian Journal of Pediatrics (2021) 47:1 27. Benini F, Trapanotto M, Spizzichino M, Lispi L, et al. Hospitalization in
children eligible for palliative care. J Palliat Med. 2010;13:711–7. 27. Benini F, Trapanotto M, Spizzichino M, Lispi L, et al. Hospitalization in
children eligible for palliative care. J Palliat Med. 2010;13:711–7. . Benini F, Trapanotto M, Spizzichino M, Lispi L, et al. Hospitalization 28. Connor SR, Downing J, Marston J. Estimating the global need for palliative
care for children: a cross- sectional analysis. J Pain Symptom Management. 2017;53(2):171–7. 29. Nasbitt J. Megatrends, grand central pub; 1982. 29. Nasbitt J. Megatrends, grand central pub; 1982. 30. Cadeddu A, D'Aloja E, Faa G, Fanos V, Granese A, Rutelli P. Qualità e sanità:
un dialogo per l'umanizzazione. Milan: Franco Angeli Editore; 2008. Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations. Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations.
|
https://openalex.org/W2265454835
|
https://link.springer.com/content/pdf/10.1007%2Fs41095-015-0026-0.pdf
|
English
| null |
Variance reduction using interframe coherence for animated scenes
|
Computational visual media
| 2,015
|
cc-by
| 4,839
|
1
Institute of Software, Chinese Academy of Sciences,
Beijing 100190, China. E-mail: P. Zhou, zhoupeng20001
@sina.com ( ); Y. Chen, chenyy@ios.ac.cn.
Manuscript received: 2015-10-23; accepted: 2015-10-29 Peng Zhou1 ( ), Yanyun Chen1 Peng Zhou1 ( ), Yanyun Chen1 c⃝The Author(s) 2015. This article is published with open access at Springerlink.com systems based on pure Monte Carlo methods to
produce a noise-free image. Although there are
some caching techniques such as irradiance caching
and photon mapping which can provide smooth
images, they usually suffer from flickering problems
because of bias. Despite having been studied for
decades, variance reduction is still an open area full
of challenges. Abstract In an animated scene, geometry and lighting
often change in an unpredictable way. Rendering
algorithms based on Monte Carlo methods are usually
employed to precisely capture all features of an
animated
scene. However,
Monte
Carlo
methods
typically take a long time to produce a noise-free
image. In this paper, we propose a variance reduction
technique for Monte Carlo methods which exploits
coherence between frames. Firstly, we introduce a
dual cone model to measure the incident coherence
intersecting camera rays in object space. Secondly, we
allocate multiple frame buffers to store image samples
from consecutive frames. Finally, the color of a pixel
in one frame is computed by borrowing samples from
neighboring pixels in current, previous, and subsequent
frames. Our experiments show that noise is greatly
reduced by our method since the number of effective
samples is increased by use of borrowed samples. In this paper,
we propose a novel variance
reduction technique for Monte Carlo methods used
to render animated scenes. We use a dual cone
model to measure the coherence between radiance
samples centered at the intersection of the camera
ray with the scene. Coherence is a metric used to
describe the similarity of radiance samples, which
enables a pixel to reuse samples from its neighboring
pixels with a suitable weight. Sample reuse happens
over a sequence of consecutive frames instead of a
single frame. Thus, multiple frame buffers must be
allocated to store the samples (instead of color) from
each frame. The contribution of our work includes
two parts. Firstly, we introduce a scheme for inter-
frame sample reuse. Secondly, we provide a criterion
to avoid using invalid samples. Keywords
physically based rendering; Monte Carlo
methods; ray tracing; global illumination;
animation 1
Introduction Physically
based
animation
has
widespread
applications in the movie industry, architectural
design, and visualization. In computer graphics,
Monte Carlo methods are the most practical way to
render animated scenes. However, the convergence
of Monte Carlo methods is inherently slow—it
is typically a time consuming task for rendering 1
Institute of Software, Chinese Academy of Sciences,
Beijing 100190, China. E-mail: P. Zhou, zhoupeng20001
@sina.com ( ); Y. Chen, chenyy@ios.ac.cn.
Manuscript received: 2015-10-23; accepted: 2015-10-29 Variance reduction using interframe coherence for animated
scenes Peng Zhou1 ( ), Yanyun Chen1 Computational Visual Media
DOI 10.1007/s41095-015-0026-0 Computational Visual Media
DOI 10.1007/s41095-015-0026-0 Computational Visual Media
DOI 10.1007/s41095-015-0026-0 Computational Visual Media
DOI 10.1007/s41095-015-0026-0 Vol. 1, No. 4, December 2015, 343–349 Research Article Research Article 2
Related work Traced global illumination is mainly used to
produce physically based images in an unbiased
way
for
animated
scenes
with
moving
objects
and dynamic lighting. Refs. [1] and [2] seamlessly
integrate photon mapping and bidirectional path
tracing into a more robust combined rendering
algorithm
via
multiple
importance
sampling,
overcoming shortcomings of traditional bidirectional
path tracing. Ref. [3] proposed gradient-domain
rendering for Monte Carlo image synthesis. They Peng Zhou, Yanyun Chen 344 found
that
estimating
image
gradients
is
also
possible using standard Monte Carlo algorithms,
and
furthermore,
even
without
changing
the
sample
distribution,
this
leads
to
significant
error reduction. Ref. [3] introduced a bidirectional
formulation for light transport and a set of weighting
strategies to significantly reduce the bias in VPL
(virtual point light)-based rendering algorithms. Because of the need to be unbiased, the samples must
be used independently, which limits the possibility
for
reuse
of
samples. Ref. [4]
used
statistical
dependency between the outputs and inputs of a
rendering system to reduce the importance of the
sample values affected by Monte Carlo noise when
applying an image-space, cross-bilateral filter. This
removes only the noise caused by randomness but
preserves important scene details. Our method can
help to greatly improve the performance of these
methods by exploiting redundant information from
samples with coherence in object space. suppress the errors, so that the images for different
frames often have inconsistent color, making the
animation flicker. Our method does not suffer from
a flickering problem, as we use a dual cone model to
give each sample a reasonable reuse weight and rate. Interactive ray tracing focuses on optimizing
acceleration structure such as grids, kd-trees, and
BVHs
(bounding
volume
hierarchy). Ref. [11]
proposed
interactive
frustum-bounded
packet
tracing on a uniform grid. Tracing performance can
be accelerated by a factor of 10 by incrementally
computing the overlap of the frustum with a slice of
grid cells. Ref. [12] proposed a method to efficiently
extend refitting for animated scenes with tree
rotations. Ref. [13] proposed a ray-space hierarchy
method, which can handle truly dynamic scenes
without the need to rebuild or update the scene
hierarchy. Ref. [14] proposed a method to selectively
restructure the BVH for ray tracing dynamic scenes. These methods optimize a different part of the
rendering pipeline from our method, so they play a
complementary role to our method in the rendering
of animated scenes. 2
Related work Cached global illumination is mainly used
to produce smoothed physically based images for
static scenes by caching and interpolating results
of lighting computations. Ref. [5] observed that
indirect irradiance typically varies smoothly. A small
number of sample points is sufficient to calculate
and cache the indirect diffuse irradiance while
keeping the error under a given threshold. Ref. [6]
introduced second order gradients (Hessians) as
a principled error bound for the irradiance; this
approach significantly outperforms the classic split-
sphere heuristic. Ref. [7] first introduced photon
mapping, in which a spatial structure called a
photon map is used to store and estimate indirect
lighting at different locations within the scene. Ref. [8] introduced progressive photon mapping
where the global illumination solution becomes
increasingly accurate and can be visualized to
provide progressive feedback. Ref. [9] introduced
a new formulation of progressive photon mapping
based on a probabilistic approach. Ref. [10] proposed
reversed photon mapping which spreads camera
importance into the scene and evaluates photon
contributions by lookup in the importance map. The interpolating step required inevitably introduces
bias into the image and the cache has to be reused
excessively to pay off. The filtering technique
employed
by
these
methods
cannot
completely 3
Measuring incident coherence using a
dual cone The light transport at a point in object space is
described by the rendering equation: Lo(x,ωωωo) =
Z
Ω
Li(x,ωωωi)f(x,ωωωo,ωωωi)V (x,ωωωi) cos θdωωω
(1) (1) where Lo is the outgoing radiance leaving shading
point x in direction ωωωo, Li is the incident radiance
arriving at x from direction ωωωi, f is the bidirectional
reflectance distribution function (BRDF), and V is
the visibility between the light source and shading
point. The goal of the dual cone model is to provide a tool
for measuring the similarity of lighting conditions
for different locations. The model assumes that all
light sources have non-zero projected area on the
illuminated surface and have constant intensity. For
a given light source, lighting coherence between two
points can be measured as the size of the solid
angle through which the light source is visible from
both locations. Precisely computing the solid angle
for shared visibility can be quite expensive because
the projected shape on the unit sphere can be very 344 344 Variance reduction using interframe coherence for animated scenes 345 blue region in Fig. 1) from the sector: complex. To make the computation practical, in the
dual cone model the projected shape of the visible
part of the light source on the unit sphere is idealised
as a disk that subtends the same solid angle as the
original shape. Soverlap
=
2(Ssector −Striangle)
Ssector
=
πr2α/2π
Striangle
=
d
2
s
r2 −
d
2
2
α
=
2 arccos d
2r Given a location x, the primary cone is defined by
three factors: (i) the direction ωωω to the light source,
(ii) the solid angle A subtended by the light source,
and (iii) the distance R to the light source, which we
call the light source distance. The geometry is shown
in Fig. 1. The secondary cone is constructed for any
other location x+∆x for whose similarity of lighting
conditions we wish to compare with location x. Note
that the secondary cone may not be placed exactly
at x + ∆x. The two cones have exactly the same
height and size, and are parallel to each other. The
metric of similarity is given by the ratio of the area
of the overlapping region to the surface area at the
base of the cone. To be specific, the metric is given
by C = Soverlap/Sdisk, where Soverlap is the overlap
area (the yellow region in Fig. 3
Measuring incident coherence using a
dual cone 1) and Sdisk is the area
of the disk at the cone base. Soverlap consists of two
symmetrical bows, each of which can be computed
by subtracting the area of inner triangle Striangle (the The relationship between half cone angle β and the
solid angle A subtended by the basal disk can be
computed as The relationship between half cone angle β and the
solid angle A subtended by the basal disk can be
computed as A
=
Z 2π
0
Z β
0
sin θdθdφ = 2π(1 −cos β)
β
=
arccos
1 −A
2π
β
=
arccos
1 −A
2π
The overlap ratio C is finally given by The overlap ratio C is finally given by C
=
Soverlap
Sdisk
= 2(Ssector −Striangle)
πr2 C
=
Soverlap
Sdisk
= 2(Ssector −Striangle)
πr2
=
2
π(arccos g −g
p
1 −g2)
(2) (2) π
where g = d/(2r). where g = d/(2r). The projected distance d is given by d = ||∆
∆
∆x −
(ωωω ·∆
∆
∆x)ωωω||, and the disk radius is r = R tan β, which
lets us compute g as p
g
g = ||∆
∆
∆x −(ωωω · ∆
∆
∆x)ωωω||
2R tan β
(3) (3) R
d
r
h
r
d/2
x
x+Dx
R
Primary
Secondary
d R
d
r
h
r
d/2
x
x+Dx
R
Primary
Secondary
d
Fig. 1
The dual cone model. The primary cone is defined by
location x, direction ωωω, solid angle A, and light source distance R. The secondary cone is constructed by moving the primary cone along
vector d, where d is the projected distance between x and x + ∆x in
the plane defined by the base of the primary cone. Figure 2 shows the overlap ratio C for different
lighting environments. Clearly, C increases with
the solid angle A (4π/30, 4π/150, and 4π/1500)
subtended by the light source, and the light source
distance R (1, 2, 4, 8, and 16). It decreases with
the projected distance d. This is consistent with
our expectation: smaller visible light sources casts
harder shadows, and larger light source distances
results in softer shadow. Fig. 1
The dual cone model. The primary cone is defined by
location x, direction ωωω, solid angle A, and light source distance R. 3
Measuring incident coherence using a
dual cone The secondary cone is constructed by moving the primary cone along
vector d, where d is the projected distance between x and x + ∆x in
the plane defined by the base of the primary cone. 4
Setup lighting environment In the first line, xi is the i-th path sample for Monte
Carlo sampling, L(xi) is the incident radiance, f(xi)
is the BRDF, cos θi is a geometric factor, and p(xi)
is probability density function (PDF). In the second
line xij is the j-th neighbor of xi. L(xij), f(xij),
cos θij, and p(xij) have already been evaluated for
xij in a previous pass. The final value of xi is
evaluated by borrowing sampled values from all its
neighbors (instead of completely tracing a path). Cj gives a weight indicating similarity of lighting
conditions between xi and xij. M
X
k=1
Cik sums the
weights from all neighbors of xi. The accuracy of
the dual cone approach is approximately equivalent
to a final gathering process with M rays per pixel,
but only involves the cost of tracing one ray. After
eliminating invalid neighbors, the overlap ratio for
valid neighbors is computed using Eq. (2). The local light source is usually modeled by a
polygon, and in practice, its intensity is specified
by a constant value or a texture map. Local lights
whose intensity is determined by a texture map can
processed as image based lights as described above. Local lights with constant intensity do not need
any change. The approach here demonstrates an
alternative for direct lighting, but for small area light
sources we still recommend the traditional approach
which samples light sources, since it is more efficient
than sampling the BSDF (bidirectional scattering
distribution function). 4.2
Exploiting inter-frame coherence Since the coherence considered is in object space,
camera motion does not change the coherence, so
it is possible to re-project samples from a previous
frame to current frame. However, the camera ray
of one frame may hit a different surface from the
ray of another frame, so visibility must be rechecked
to validate samples from such previous frames. Furthermore, the object and light can also be in
motion, so the visibility of secondary rays associated
with valid samples must also be rechecked. 4
Setup lighting environment The dual cone model ignores the impact of the
incident
light’s
intensity. When
comparing Fig. 2
Overlap ratio for different light source distance R, solid angle A, and projected distance d. Fig. 2
Overlap ratio for different light source distance R, solid angle A, and projected distance d. 346 Peng Zhou, Yanyun Chen =
1
N
N
X
i=1
1
PM
k=1 Cik
M
X
j=1
Cj
L(xij)f(xij) cos θij
p(xij)
=
1
N
N
X
i=1
1
W
M
X
j=1
(Cj · Fij)
(4) two radiance samples, it assumes that the lighting
conditions are constant. The original lighting model
must be converted to a group of area lights with low
radiance variance, in order to meet the assumption. =
1
N
N
X
i=1
1
W
M
X
j=1
(Cj · Fij)
(4) (4) For image based lighting, such as an environment
map, the image is subdivided into an axis-aligned
kd-tree by recursively cutting the light intensity in
half. The leaves of the kd-tree are used as area lights,
whose intensities are approximated by the average of
all pixels each contains. In the worst case, every pixel
will be used as an area light. For extremely smooth
images, the entire image may be used as a single area
light. In the first line, xi is the i-th path sample for Monte
Carlo sampling, L(xi) is the incident radiance, f(xi)
is the BRDF, cos θi is a geometric factor, and p(xi)
is probability density function (PDF). In the second
line xij is the j-th neighbor of xi. L(xij), f(xij),
cos θij, and p(xij) have already been evaluated for
xij in a previous pass. The final value of xi is
evaluated by borrowing sampled values from all its
neighbors (instead of completely tracing a path). Cj gives a weight indicating similarity of lighting
conditions between xi and xij. M
X
k=1
Cik sums the
weights from all neighbors of xi. The accuracy of
the dual cone approach is approximately equivalent
to a final gathering process with M rays per pixel,
but only involves the cost of tracing one ray. After
eliminating invalid neighbors, the overlap ratio for
valid neighbors is computed using Eq. (2). 346 4.3
The rendering framework Algorithm - Progressive refinement
1
Subdivide environment into N directional lights
2
Build light tree
3
Allocate frame buffer queue of size B
4
for every B frames in animation
5
Clear frame buffer queue
6
for i = 1 to n do
7
Run Pass 1
8
Run Pass 2
9
Accumulate image samples from Pass 2
10
into frame buffer
11
end for The dual cone model rendering process has two
passes. 1) Sampling pass. For each pixel in each frame,
we use a single gathering ray to sample radiance for
each pixel, and record the result to an image cache. The record includes:
radiance/direction/length of
sampling ray, solid angle subtended by visible light,
position and surface normal at the camera ray
intersection. 1) Sampling pass. For each pixel in each frame,
we use a single gathering ray to sample radiance for
each pixel, and record the result to an image cache. The record includes:
radiance/direction/length of
sampling ray, solid angle subtended by visible light,
position and surface normal at the camera ray
intersection. We
can
simply
run
the
rendering
algorithm
multiple times, and accumulate image samples from
each iteration into the frame buffer. The result in the
frame buffer will gradually converge to the correct
value by the nature of the Monte Carlo process. The
frame sequence in the animation is split into frame
blocks with B frames. Samples are shared among
all frames in the same block, and coherence between
blocks is ignored. The total memory footprint is B
frame buffers with some extra bytes per pixel. 2) Rendering pass. For each pixel in each frame,
records from N × N pixel blocks centered at the
current pixel position in every frame are used to
determine the final color. First, we trace a ray
for the current pixel to find an intersection i. Then we validate the borrowed record, by checking
normal difference, distance, and visibility. Next, we
compute the overlap ratio C according to Eq. 4.3
The rendering framework (2) to
weight the lighting similarity between intersection i Algorithm - Pass 1
1
for each frame a in frame buffer queue do
2
for each pixel i in frame a do
3
Trace one camera ray from i into scene
4
Record the position and normal of intersection
5
Trace one ray based on BSDF
6
if ray missed, evaluate radiance using
7
chosen directional light, record solid angle
8
else ray is occluded
9
set radiance to zero, record solid angle
10
Record radiance, ray direction and ray length
11
end for
Algorithm - Pass 2
1
for each frame a in frame buffer queue do
2
for each pixel i in frame a do
3
Trace one camera ray to find intersection
4
for each frame b in frame buffer queue do
5
for each pixel j in N × N block centered at i do
6
Get record from pixel j in frame b
7
Check normal, distance and visibility
8
Ignore invalid record
9
Compute C for i and j using Eq. (2)
10
Update pixel i using Eq. (5)
11
end for
12
Save contribution of pixel i in frame a
13
end for
14
end for 4.1
Exploiting intra-frame coherence For each pixel in an image, camera ray samples
from nearby pixels are borrowed. The assumption
lying behind this is that nearby pixels have a high
probability of representing close by points in the
object. However, in some cases this assumption
may fail. First, adjacent camera rays may hit two
different objects that are far away from each other. Second, two camera rays may hit two surfaces whose
orientations are very different. In the first case, we
project the pixel block centered at the current pixel
into object space and use the projected area as a
search region for valid neighboring pixels. To cover
the second case, we decide that if the cosine of the
angle between the two surface normals is no less than
0.9, then the two pixels can borrow sample from each
other. In order to share samples between frames, we
render multiple frames at the same time. To store
consecutive frames, a frame buffer queue is allocated. Frames are arranged in ascending order. The queue
size is left as a parameter for the user. One-sample-
per-pixel rendering is performed for each frame in
the queue. A sample from one frame can be reused
by any frame in the queue. The rendering process
is iteratively performed until the required number of
samples per pixel is met. Equation (4) gives the final contributions to a pixel
after borrowing samples in the current frame. L = 1
N
N
X
i=1
L(xi)f(xi) cos θi
p(xi) Equation (5) gives the final contributions to a pixel 346 Variance reduction using interframe coherence for animated scenes 347 by borrowing samples from multiple frames. and the intersection cached in the record of the j-
th neighbor. Finally we sum the weighted radiance
values from records according to Eq. (5) to give the
final pixel color. L
=
1
N
N
X
i=1
1
W
M
X
j=1
(Cj · Fij)
=
1
N
N
X
i=1
1
W
B
X
b=1
Mb
X
j=1
(Cbj · Fbij)
(5) (5) 4.4
Progressive refinement The two-pass rendering described above is essentially
an approximated version of final gathering with one
sample per pixel, while the number of gathering
rays is determined by the number of available pixel
records. It is a special case of Eq. (5) where N = 1. where M = PB
b=1 Mb. In this modified version of
Eq. (4), B is the size of the frame buffer queue, and
Mb is the number of samples borrowed from frame b. 5
Results and discussion We first applied our method to render a scene
showing the main stadium for the 29th Olympic
Games, consisting of 114,000 polygons. The rendered
result is shown in Fig. 3. The richness of lighting
effects is correctly captured with only 32 samples per
pixel (SPP). This result shows the robustness of our
method for a scene with complex geometry. We then applied our method to render a working
loader which consists of 59,000 polygons, as shown in
Fig. 4. In this scene, the camera and objects are both
in motion. By sharing samples between multiple
consecutive frames, our method produces a smooth
image with only 32 samples per pixel. Plausible
shadows are ensured by rechecking visibility. We also compared our method to image based
filtering and irradiance cache methods. All rendered 348 Peng Zhou, Yanyun Chen the same scene using 32 samples per pixel. The
result is shown in Fig. 5. The image based method
can smooth the image by averaging the contribution
of nearby pixels, but it also blurs details. Using
an irradiance cache smooths the image by reusing
irradiance values in object space, but it has low
frequency artifacts where the sampling rate is not
high enough. Our method borrows and reweighs
samples from neighborhoods without introducing
blurs and artifacts. 3 GB memory. The performance data for all the
experimental results are given in Table 1. Variance reduction using interframe coherence for animated scenes Variance reduction using interframe coherence for animated scenes 349 Table 1
Performance data
Scene
Triangles
Resolution
SPP
Pass 1
Pass 2
Total time
Indoor (Fig. 5)
19,000
720 × 480
32
11.72 s
56.80 s
70.54 s
Loader (Fig. 4)
59,000
720 × 480
32
13.93 s
53.38 s
68.99 s
Stadium (Fig. 3)
114,000
720 × 480
32
50.43 s
61.04 s
119.64 s
1024 × 576
32
75.48 s
87.01 s
171.36 s to animated scenes with both a dynamic camera
and objects. Our experimental results show that
our method can produce much smoother images
than the Monte Carlo method, using very limited
samples and memory footprint. In the future, we will
combine our work with image based noise reduction
methods such as the one proposed by Ref. [4]. [9] Knaus, C.; Zwicker, M. Progressive photon mapping:
A
probabilistic
approach. ACM
Transactions
on
Graphics Vol. 30, No. 3, Article No. 25, 2011. [10] Havran, V.;
Herzog, R.;
Seidel, H.-P. Fast final
gathering via reverse photon mapping. Computer
Graphics Forum Vol. 24, No. 3, 323–332, 2005. [11] Wald, I.; Ize, T.; Kensler, A.; Knoll, A.; Parker, S. G. Ray tracing animated scenes using coherent grid
traversal. ACM Transactions on Graphics Vol. 25, No. 3, 485–493, 2006. Open
Access
This article is distributed under the
terms of the Creative Commons Attribution License which
permits any use, distribution, and reproduction in any
medium, provided the original author(s) and the source are
credited. [12] Kopta, D.; Ize, T.; Spjut, J.; Brunvand, E.; Davis, A.;
Kensler, A. Fast, effective BVH updates for animated
scenes. In:
Proceedings of the ACM SIGGRAPH
Symposium on Interactive 3D Graphics and Games,
197–204, 2012. [13] Roger, D.; Assarsson, U.; Holzschuch, N. Whitted ray-
tracing for dynamic scenes using a ray-space hierarchy
on the GPU. In: Proceedings of the 18th Eurographics
Conference on Rendering Techniques, 99–110, 2007. 6
Conclusions In conclusion,
we have proposed a dual cone
model to reduce variance for rendering animations. In conclusion,
we have proposed a dual cone
model to reduce variance for rendering animations. Our method reduces noise by exploiting incident
coherence of radiance samples. The samples can be
shared not only within one frame, but also between
consecutive frames. The sample record must be
validated before reuse to ensure correct shadows. To
demonstrate its effectivity, we applied our method The
experiment
for
direct
lighting
using
an
environment map ran on a workstation with two
Xeon
R⃝5150 CPUs (dual core,
2.66 GHz) and (a) Path tracing
(b) Intra-frame
(c) Inter-frame
(d) Reference
Fig. 3
Rendering results for the steel structure model of a stadium: (a) path tracing result; (b) smoothed result by exploiting intra-frame
coherence; (c) smoothed result by exploiting inter-frame coherence; (d) reference image. The bottom line gives close-up detail. Images (a)–(c)
were all rendered using 32 samples per pixel. Fig. 3
Rendering results for the steel structure model of a stadium: (a) path tracing result; (b) smoothed result by exploiting intra-frame
coherence; (c) smoothed result by exploiting inter-frame coherence; (d) reference image. The bottom line gives close-up detail. Images (a)–(c)
were all rendered using 32 samples per pixel. Fig. 4
Dynamic scene of a working loader. Four views rendered by our method using 32 samples per pixel. Fig. 4
Dynamic scene of a working loader. Four views rendered by our method using 32 samples per pixel. (Path tracing)
(Bilateral filter)
(Irradiance cache)
(Our method)
(Reference)
Fig. 5
Performance comparison for different methods. Top left: path tracing result. Image A applies bilateral image filtering to the path
tracing result. Image B: result using an irradiance cache. Image C: result using our method. Image D: reference. (Reference) Fig. 5
Performance comparison for different methods. Top left: path tracing result. Image A applies bilateral image filtering to the path
tracing result. Image B: result using an irradiance cache. Image C: result using our method. Image D: reference. 348 348 References [1] Georgiev, I.; Kˇriv´anek, J.; Davidoviˇc, T.; Slusallek, P. Light transport simulation with vertex connection and
merging. ACM Transactions on Graphics Vol. 31, No. 6, Article No. 192, 2012. [14] Yoon, S.-E.; Curtis, S.; Manocha, D. Ray tracing
dynamic
scenes
using
selective
restructuring. In:
Proceedings of the 18th Eurographics Conference on
Rendering Techniques, 73–84, 2007. [2] Hachisuka, T.; Pantaleoni, J.; Jensen, H. W. A path
space extension for robust light transport simulation. ACM Transactions on Graphics Vol. 31, No. 6, Article
No. 191, 2012. system. Peng Zhou is a postdoctoral fellow
in the Institute of Software, Chinese
Academy
of
Sciences. He
received
his Ph.D. degree in computer science
from Shandong University in 2012. His
research
interests
include
physically
based rendering, ray tracing acceleration
structure,
and
creating
rendering [3] Walter, B.; Khungurn, P.; Bala, K. Bidirectional
lightcuts. ACM Transactions on Graphics Vol. 31, No. 4, Article No. 59, 2012. [4] Sen, P.; Darabi, S. On filtering the noise from the
random parameters in Monte Carlo rendering. ACM
Transactions on Graphics Vol. 31, No. 3, Article No. 18, 2012. system. [5] Ward,
G. J.;
Rubinstein,
F. M.;
Clear,
R. D. A ray tracing solution for diffuse interreflection. In:
Proceedings of the 15th Annual Conference on
Computer Graphics and Interactive Techniques, 85–
92, 1988. Yanyun
Chen
is
a
professor
in
the
Institute
of
Software,
Chinese
Academy of Sciences. He received
his
Ph.D. degree
in
January
2000. His research interests include photo-
realistic rendering, non-photo-realistic
rendering, computer animation, virtual
reality, and textures. [6] Schwarzhaupt, J.; Jensen, H. W.; Jarosz, W. Practical
Hessian-based error control for irradiance caching. ACM Transactions on Graphics Vol. 31, No. 6, Article
No. 193, 2012. [7] Jensen, H. W. Global illumination using photon maps. In:
Proceedings of the Eurographics Workshop on
Rendering Techniques, 21–30, 1996. Other papers from this open access journal are available free
of charge from http://www.springer.com/journal/41095. To submit a manuscript,
please go to https://www. editorialmanager.com/cvmj. [8] Hachisuka, T.; Ogaki, S.; Jensen, H. W. Progressive
photon mapping. ACM Transactions on Graphics Vol. 27, No. 5, Article No. 130, 2008.
|
https://openalex.org/W4380722139
|
https://zenodo.org/records/8039777/files/209-212.pdf
|
Indonesian
| null |
TA'LIM JARAYONIDA BOSHLANG'ICH SINFLARDA TEXNOLOGIYA FANINI O'QITISH MUAMMOSINING YORITILISH MAZMUNI
|
Zenodo (CERN European Organization for Nuclear Research)
| 2,023
|
cc-by
| 2,349
|
Foydalanilgan adabiyotlar 1. Oʻzbekiston Respublikasining yangi tahrirdagi «Ta’lim toʻgʻrisida»gi Qonuni. Toshkent. 2020-
yil, 23-sentabr. 1. Oʻzbekiston Respublikasining yangi tahrirdagi «Ta’lim toʻgʻrisida»gi Qonuni. Toshkent. 2020-
yil, 23-sentabr. y
2. Oʻzbekiston Respublikasi prezidenti Sh.Mirziyoyevning 2019-yil 9-iyuldagi«Matematika
ta’limi va fanlarini yanada rivojlantirishni davlat tomonidan qo‘llab-quvvatlash chora-tadbirlari
to‘g‘risida»gi qarori. y
2. Oʻzbekiston Respublikasi prezidenti Sh.Mirziyoyevning 2019-yil 9-iyuldagi«Matematika
ta’limi va fanlarini yanada rivojlantirishni davlat tomonidan qo‘llab-quvvatlash chora-tadbirlari
to‘g‘risida»gi qarori. g q
3. L. O‘rinboyeva va b 1-sinf matematika kitobi. Toshkent-2021. g q
3. L. O‘rinboyeva va b 1-sinf matematika kitobi. Toshkent-2021. y
4. L. O‘rinboyeva va b 2-sinf matematika metodik qo‘llanma kitobi. Toshkent-2021. y
4. L. O‘rinboyeva va b 2-sinf matematika metodik qo‘llanma kitobi. Toshkent-2021. 5. Jumayev M.E. Bolalarda matematika tushunchalarni shakllantirish nazariyasi.-T.: «Ilm-Ziyo»,
005 5. Jumayev M.E. Bolalarda matematika tushunchalarni shakllantirish nazariyasi.-T.: «Ilm-Ziyo»,
2005, 2005, Natijada, oldingi asrning 60-yillari boshidan AQShda so‘ngra boshqa g‘arb
mamlakatlarida ishchanlik o‘yinlari qo‘llanila boshlandi. Ishchanlik o‘yinlari tadqiqotchilari bu
usulni eng asosiy, samarali va tejamli ta’lim metodlaridan iborat deb ta’kidlaganlar. Natijada, oldingi asrning 60-yillari boshidan AQShda so‘ngra boshqa g‘arb
mamlakatlarida ishchanlik o‘yinlari qo‘llanila boshlandi. Ishchanlik o‘yinlari tadqiqotchilari bu
usulni eng asosiy, samarali va tejamli ta’lim metodlaridan iborat deb ta’kidlaganlar. -muammoli vaziyat yaratish; -g‘oyalarni shakllantirish; -eng yaxshi g‘oyalarni tekshirish, baholash va tanlash. g y
g
y
Didaktik o‘yin texnologiyalari orqali amalga oshiriladigan ayrim noana’naviy mashg‘ulot
shakllari: g y
g
y
Didaktik o‘yin texnologiyalari orqali amalga oshiriladigan ayrim noana’naviy mashg‘ulot
shakllari: Ishchanlik o‘yini darsi- mashg‘ulot mavzusi bo‘yicha masalalarni hal etish jarayonida
bolalarning faol ishtirok etishini ta’minlash orqali yangi bilimlarni o‘zlashtirish mashqi. Rolli o‘yin mashg‘uloti - mashg‘ulot mavzusi bo‘yicha masalalarni o‘rganishda bolalarga
oldindan ma’lum rollarni taqsimlash va mashg‘ulot jarayonida shu rolni bajarishlarini tashkil
etish asosida bilimlarni mustahkamlash darsi. Teatrlashtirilgan mashg‘ulot – mashg‘ulot mavzusi bilan bog‘liq sahna ko‘rinishlari
tashkil etish orqali mashg‘ulot mavzusi bo‘yicha chuqur, aniq ma’lumotlar berish darsi. Kompyuter mashg‘uloti - tegishli mashg‘ulot turi bo‘yicha mashg‘ulot mavzusiga doir
kompyuter materiallari (multimedia, virtual o‘quv kursi va shu kabilar) asosida o‘tiladigan
dars. Yarmarka mashg‘uloti – mashg‘ulot mavzusini bo‘laklar bo‘yicha oldindan o‘zlashtirgan
bolalarning o‘zaro muloqot asosida guruhga qiziqarli tushuntirish orqali o‘tiladigan mashg‘ulot. Integral (integratsiyalashgan) mashg‘ulot - bir nechta mashg‘ulot turlariga doir va
integratsiyalash uchun qulay bo‘lgan mavzular bo‘yicha tashkil qilingan mashg‘ulot bo‘lib,
bolalarning turli mashg‘ulot turlarga qiziqishlarini orttirib, ta’lim jarayonidagi faolliklarini
ta’minlaydi. TA’LIM JARAYONIDA BOSHLANG‘ICH SINFLARDA TEXNOLOGIYA FANINI
O‘QITISH MUAMMOSINING YORITILISH MAZMUNI TA’LIM JARAYONIDA BOSHLANG‘ICH SINFLARDA TEXNOLOGIYA FANINI
O‘QITISH MUAMMOSINING YORITILISH MAZMUNI TA’LIM JARAYONIDA BOSHLANG‘ICH SINFLARDA TEXNOLOGIYA FANINI
O‘QITISH MUAMMOSINING YORITILISH MAZMUNI Hojiyeva Nasiba Bahodirovna Annotatsiya. Ushbu ta’lim jarayonida boshlang‘ich sinflarda texnologiya fanini o‘qitish muammosining
yoritilishi mazmuni yoritib berilgan. O‘quvchilarni boshlang‘ich sinfdagi texnologiya ta’limining asosiy vazifalari
mehnatga axloqiy va ruhiy tayorlash, o‘quvchilarni boshlang‘ich politexnik bilimlar bilan qurollantirish, mehnatga
amaliy tayorlashlardan iboratdir. Kalit so‘zlar: texnologiya darslari, kreativ, kompetensiya, ishlab chiqarish, milliy dastur, rivojlantirish,
zamonaviy texnologiya. y
g y
Kirish: O‘zbekistonning mustaqillikka erishgani, har tomonlama jadallik bilan rivojlanib
borayotgani, Respublikamizda amalga oshirilayotgan siyosiy, iqtisodiy va ma’naviy o‘zgarishlar
jamiyatimiz ijtimoiy hayotida tub burilish boshlab berdi. Darhaqiqat, Prezidentimiz
“O‘zbekistonni rivojlangan mamlakatga aylantirishni maqsad qilib qo‘ygan ekanmiz, bunga
faqat jadal islohotlar, ilm-ma’rifat va innovatsiya bilan erisha olamiz. Buning uchun, avvalambor,
tashabbuskor islohotchi bo‘lib maydonga chiqadigan, strategik fikr yuritadigan, bilimli va
malakali yangi avlod kadrlarini tarbiyalashimiz zarur. Shuning uchun ham 209 bog‘chadan boshlab oliy o‘quv yurtigacha – ta’limning barcha bo‘g‘inlarini isloh qilishni
boshladik. Nafaqat yoshlar, balki butun jamiyatimiz a’zolarining bilimi, saviyasini oshirish uchun
avvalo ilm-ma’rifat, yuksak ma’naviyat kerak. Ilm yo‘q joyda qoloqlik, jaholat va albatta, to‘g‘ri
yo‘ldan adashish bo‘ladi” deb ta’kidlab o‘tdilar. Texnologiya darslari o‘quvchilarga tafakkur, diqqat, ijodiy tasavvur etish, mehnatga
muhabbatini rivojlantirishda imkom beradi. Shuningdek texnologiya darslari o‘quvchilarni
dunyoqarashlarini rivojlantirishga zamin yaratadi. Undan tashqari texnologiya ta’limi
o‘quvchilarda aqliy va jismoniy mehnat turlari, mehnat jarayonlari haqida keng tushuncha
hosil qilish, mehnatga oid ko‘nikma va malakalarni rivojlantirish kabi ta’limiy maqsadlarga
xizmat qiladi. Hozirda umumta’lim maktab boshlang‘ich sinflar oldiga qo‘yiladigan talablar ortib
bormoqda. Bu talablar tabiat va jamiyat talablarini bilish maqsadida ta’lim mazmuni fan
texnikaning eng yangi yutuqlari bilan boyitishga, ta’lim tarbiya jarayonida texnologiya
ta’limining ro‘lini o‘stirish va bolalarni tarbiyalash uchun ota-onalarning,o‘qituvchilarning
ma’suliyatini o‘stirishga qaratilgan. Bu o‘z navbatida texnologiya ta’limi oldiga murakkab
ma’suliyatli vazifalar qo‘yadi. Vujudga kelgan imkoniyatlardan mehnat o‘qituvchisi kasbini har
tomonlama rivojlanishi uchun to‘liq imkoniyatini beradi. Mehnat ta’limi mutaxassis tayyorlashni
o‘z maqsadi qilib qo‘ymaydi, balki u o‘quvchi shaxsini rivojlantirish uchun vosita hisoblanadi. q
q
q
y
y
q
j
Boshlang‘ich sinflardagi texnologiya ta’limi tarbiyasi va ma’lumoti, maktabda mehnat
asosini namoyon bo‘lishi va uning o‘zagi bo‘lib, u bolalarning bilimi, ishlab chiqarish faoliyati,
o‘zaro yordam ko‘rsatish, estetik va jismoniy rivojlantirish jarayonidagi mehnat kuchi haratlarini
qamrab oladi. Boshlang‘ich ta’limi texnologiya ma’lumotining tarkibiy qismi hisoblanadi. Ta’lim
jarayonida yigit va qizlar mehnatining nazariy va amaliy hamda samarali, eng oddiy asosiy
negizlari va tarkibiy qismlarini, umumlashgan mehnat malakalarini bilish va amaliy faoliyat
usullarini egallashga qaratilgan. g
g q
g
1932-yilda boshlang‘ich va o‘rta maktablarning o‘quv dasturlari va tartibi to‘g‘risidagi
qarorida maktablarda fan asoslarini fanlardan foydalanib o‘tish ishlab chiqildi. Hojiyeva Nasiba Bahodirovna Boshlang‘ich sinfda o‘quvchilarning amaliy faoliyatlariga asosiy e’tibor beriladi, bunda
o‘quv vaqtining 80-85 % ajratiladi. Boshlang‘ich sinfda o‘quvchilarning amaliy faoliyatlariga asosiy e’tibor beriladi, bunda
o‘quv vaqtining 80-85 % ajratiladi. Xulosa Mamlakatimiz kelajagini qurishda eng katta berilayotgan e’tibor maktablarimizning
infratuzilmasi, ta’limda sifat va samaradorlikni oshirishga qaratilmoqda. Bunday mas`uliyatli
vazifani amalga oshirishda boshlang`ich sinf o`qituvchilarning ham o‘rni, roli benihoya katta. Shuning uchun bugungi kun o`qituvchisi zamonaviy texnologiyalardan foydalana olishi va ayni
damda ushbu texnologiyalarni amaliyotga tadbiq eta bilishi juda muhimdir. O`quvchilarning fikr
doirasi, ongi dunyoqarashini o`stirish, ularni mustaqil fikrlaydigan, o`z fikr-mulohazalarini to‘liq
bayon eta oladigan, xalqaro tadqiqotlarda munosib o‘rinlarni egallay oladigan, erkin
ishritirokchiga aylantirish, bugungi zamon talabining eng muhim masalalaridan biridir. Buning
uchun esa ular ilm olmog‘i, tinmay o‘qishi, o‘z ustida ishlashi va eng asosiysi, yaqin do`sti kitob
bo`lmog`i joizdir. g
j
Boshlang‘ich sinflarda badiiy asarlarni o‘qitish, tahlil qilish, kompozitsiyani yaxlit
anglagan holda matnning asosiy ma’nosini yoritib berish, asarda ifodalangan qahramonlar,
ularning hayot tarzi, voqea tasvirlangan joy nomlari ifodalab nazariy va ilmiy jihatdan to‘g‘ri
asoslanishi, o‘quvchida shu asarni yaxlit tushuna olishi, mulohaza yuritishi asarning mazmunini
anglashiga yordam beradi. Hikoyani o‘qitishdagi nazariy asoslar shuningdek, bugungi kundagi
hikoya va uni o‘qitishdagi yondashuvlar, olimlarning hikoyani o‘qitishdagi turli metodik
izlanishlari bu bobda o‘z aksini topgan. Hikoyaning kelib chiqish tub asoslari, boshlang‘ich ta’lim
darsliklaridagi mazmunan boy ayrim hikoya yozgan adiblarimizning hikoyalaridan namunalar
asoslanib keltirilgan. Umuman olganda, adabiy matn ustida ishlash, tahlil qilish va bu borada
ilgari surilgan fikrlar, ularning nazariy asoslari darsliklardagi topishmoq, qissa, ertak, hikoya,
she’r kabi nasriy va she’riy uslubda yozilgan asarlar, ularning boshlang‘ich sinf darsliklaridagi
qamrovi, farqli jihatlari, o‘qitish metodikasidagi umumiy fikrlar haqida so‘z yuritilgan. 1. O‘zbekiston Respublikasi Prezidenti Sh.Mirziyoyevning 2020 yil 24 yanvarda Oliy Majlisg
tnomasida so‘zlagan nutqi. j
g
q
2. Milliy o‘quv dasturi “Texnologiya” j
g
q
2. Milliy o‘quv dasturi “Texnologiya” 3. Z.D.Rasulova, Sh.H.Quliyeva, A.R.Jo‘rayev “Texnologiya fanini o‘qitish metodikasi” Buxoro-
2020 4. Sanaqulov.H.R. N.Qambarov “Mehnat va uni o‘qitish metodikasi” ma’ruza matni, Namangan,
2013 4. Sanaqulov.H.R. N.Qambarov “Mehnat va uni o‘qitish metodikasi” ma’ruza matni, Namangan,
2013 5. Amonov U. S. O ‘qish darslarida maqol janridan foydalanish usullari va ahamiyati: DOI:
10.53885/edinres. 2021.53. 34.124 US
O
A
Ek
l
h
k
h
k
l
k 6. O.R.Avezov. Ekstrennaya psixologicheskaya pomosh v ekstremalnix situatsiyax. tegrativnoy psixologii 4 (No. 21), 34-37. 7. Nigora
Adizova
Baxtiyorovna. 3-4-sinf
ona
tili
darslarida
qoʻllanadigan
tayanch
kompetensiyalar. Journal of Advanced Research and Stability. Volume: 02 Issue: 01 | 2022 7. Nigora
Adizova
Baxtiyorovna. 3-4-sinf
ona
tili
darslarida
qoʻllanadigan
tayanch
kompetensiyalar. Journal of Advanced Research and Stability. Volume: 02 Issue: 01 | 2022 p
y
y
8. Rakhmonovich, Adizov Bakhtiyor, and Nodira Adizova Bakhtiyorovna. Hojiyeva Nasiba Bahodirovna O‘quv rejasida
boshqa o‘quv fanlari bilan bir qatorda mehnat ta’limi ham mustaqil fan bo‘lib kirdi. q
y
q
q
boshqa o‘quv fanlari bilan bir qatorda mehnat ta’limi ham mustaqil fan bo‘lib kirdi. q
q
q
q
1933-yilda maktablarda mustaqil o‘quv fani sifatidagi mehnat ta’limi tugatildi. q
q
q
q
1933-yilda maktablarda mustaqil o‘quv fani sifatidagi mehnat ta’limi tugatildi. y
q
q
g
g
1952-yilda mehnat ta’limiga jamoatchilik e’tibori maktab boshlang‘ich sinflarda ta’lim va
mehnat tarbiyasini rivojlantirishni zarurligi ko‘rsatildi. 1952-yilda mehnat ta’limiga jamoatchilik e’tibori maktab boshlang‘ich sinflarda ta’lim va
mehnat tarbiyasini rivojlantirishni zarurligi ko‘rsatildi. 1954-yilda mehnat ta’limi boshlang‘ich sinf o‘quv rejasiga o‘sib kelayotgan yosh avlodni
kamol toptirish vazifalaridan biri sifatida kiritilgan. 1954-yilda mehnat ta’limi boshlang‘ich sinf o‘quv rejasiga o‘sib kelayotgan yosh avlodni
kamol toptirish vazifalaridan biri sifatida kiritilgan. p
g
2018-yildan esa mehnat ta’limi texnologiya ta’limi bilan almashtirildi. p
g
2018-yildan esa mehnat ta’limi texnologiya ta’limi bilan almashtirildi. Sharq mutafakkirlarining pedagogik g‘oyalari uzoq o‘tmishga borib taqaladi. O‘rta
Osiyodan o‘tgan qadimiy buyuk Ipak yo‘li Xitoy, Hindiston shaharlari bilan bog‘lanib,
madaniyat va sanoatni rivojlantirishga hamda ilmga bo‘lgan qiziqishga alohoida e’tibor berishni
taqozo etadi. Bu esa o‘z navbatida boshlang‘ich sinf o‘quvchilarini mehnat qilishiga
munosabatini o‘zgartirib raqobatga arzigulik o‘z-o‘ziga ko‘rsatish orqali qishloq xo‘jaligi
maxsulotlarini ishlab chiqarishga va hunarmandchilikka va tikuvchilikka qiziqishlarini
takomillashtiradi. Texnolodiya ta’limi o‘qituvchilar rahbarligida o‘quvchilar tomonidan
bajariladigan aqliy, jismoniy harakatlar jarayonidan iborat bo‘lib, yakuniy natijalarga ularning
mehnat qurollari, vositalari va jarayonlari haqidagi bilimlarini hamda ma’lum sohadagi ishlab
chiqarish mehnatini bajarish uchun zarur amaliy ko‘nikma va malakalarni egallashlariga ongli
ravishda kasb fanlarga hamda jamiyat va shaxs farovonligini rivojlantirishga qaratilgan o‘quv
fanidir. Texnologiya ta’limining mazmuni esa shu natijalarga to‘la bog‘liqdir, ya’ni shu
vazifalarga ko‘ra belgilanadi. Masalan, ta’limni umumiy mehnat bilan birga ko‘rishni ta’minlash
zarurligi boshlang‘ich sinfdagi o‘qitish mazmuniga jumladan, mehnat ta’limi mazmuniga qator
talablar qo‘ydi: q
y
1. Boshlang‘ich sinf o‘quvchilarining faoliyatlari imkoniyatiga qarab unumli mehnatni
o‘z ichiga olish kerak . 1. Boshlang‘ich sinf o‘quvchilarining faoliyatlari imkoniyatiga qarab unumli mehnatni
o‘z ichiga olish kerak . 2. Boshlang‘ich sinf o‘quvchilarining unumli mehnati ularning o‘quv faoliyatlar Boshlang‘ich sinf o‘quvchilarining unumli mehnati ularning o‘quv faoliyatlariga, 210 tarkibiga qo‘shilib ketishlari kerak. Unumli mehnat mazmunida bolalarning yosh imkoniyatlari, mehnat ta’limi vazifalari,
qarish muhitni harakatini hisobga olishi kerak. 3. Unumli mehnat mazmunida bolalarning yosh imkoniyatlari, mehnat ta’limi
ishlab chiqarish muhitni harakatini hisobga olishi kerak. q
g
4. Maktab boshlang‘ich sinflariga ta’limning ishlab chiqarish malakasini berish bilan
tugallanishi kerak. 4. Maktab boshlang‘ich sinflariga ta’limning ishlab chiqarish malakasini berish bilan
tugallanishi kerak. Xulosa "linguistic
classification of toponyms of bukhara district." Emergent: Journal of Educational Discoveries and Lifelong
Learning (EJEDL) 2.09 (2021): 1- 8. Rakhmonovich, Adizov Bakhtiyor, and Nodira Adizova Bakhtiyorovna. "linguistic
classification of toponyms of bukhara district." Emergent: Journal of Educational Discoveries and Lifelong
Learning (EJEDL) 2.09 (2021): 1- 10. Hayitov H. A. Sharq mumtoz adabiyotida ohang talqini //Мировая наука. – 2019. – №. 8. – С. 3-5. 10. Hayitov H. A. Sharq mumtoz adabiyotida ohang talqini //Мировая наука. – 2019. – №. 8. – С. 3-5 211 11. Saidova Mohinur Jonpulatovna, Ibrahimova Mohichehra Furkat Qizi. An integrated approach
to the use of pedagogical technologies in primary school mathematics// Middle European Scientific
Bulletin. Volume 8, January 2021, 174 11. Saidova Mohinur Jonpulatovna, Ibrahimova Mohichehra Furkat Qizi. An integrated approach
to the use of pedagogical technologies in primary school mathematics// Middle European Scientific
Bulletin. Volume 8, January 2021, 174 , J
y
,
12. Ergashovna S. G. Formation of concepts about length in primary school students, developing
length measuring skills. – 2021. J
y
12. Ergashovna S. G. Formation of concepts about length in primary school students, developing
length measuring skills. – 2021. 13. Джураева С. Н., Дустова Д. С. Способы воспитания личных качеств у студентов
педагогической специальности //Academy. – 2019. – №. 6 (45). – С. 96-98 13. Джураева С. Н., Дустова Д. С. Способы воспитания личных качеств у студентов
педагогической специальности //Academy. – 2019. – №. 6 (45). – С. 96-98 y
14. O‘ktamovna X. N. O‘zbek romanlarida tarixiy inversiyani berish usullari va vositalari
//Барқарорлик ва Етакчи Тадқиқотлар онлайн илмий журнали. – 2021. – Т. 1. – №. 5. – С. 430-433. 14. O‘ktamovna X. N. O‘zbek romanlarida tarixiy inversiyani berish usullari va vositalari
//Барқарорлик ва Етакчи Тадқиқотлар онлайн илмий журнали. – 2021. – Т. 1. – №. 5. – С. 430-433. BuxDPI 2-kurs magistrantlari Kalit so‘zlar: ijtimoiy pedagogika, ijtimoiylashuv, ijtimoiy moslashuv, indivuallashuv Kalit so‘zlar: ijtimoiy pedagogika, ijtimoiylashuv, ijtimoiy moslashuv, indivuallashuv
Annotatsiya: ushbu maqolada interfaol mashqlar asosida boshlang‘ich sinf o‘quvchilarida ijtimoiy
ko‘nikmalarini shakllantirishning bir qancha yo‘llari ustida fikr-mulohazalar keltirilib o‘tiladi. j
y p
g g
j
y
j
y
notatsiya: ushbu maqolada interfaol mashqlar asosida boshlang‘ich sinf o‘quvchilarida ijtimoiy
ni shakllantirishning bir qancha yo‘llari ustida fikr-mulohazalar keltirilib o‘tiladi. Annotatsiya: ushbu maqolada interfaol mashqlar asosida boshlang‘ich sinf o‘quvch
ko‘nikmalarini shakllantirishning bir qancha yo‘llari ustida fikr-mulohazalar keltirilib o‘tiladi. Har bir shaxsning jamiyatda yashashi, uning ijtimoiy normalarga rioya qilgan holda
o`ziga o`xshash shaxslar bilan o`rnatadigan murakkab o`zaro munosabatlari va ularning ta’sirida
hosil bo`ladigan hodisalarning pedagogik tabiatini va qonuniyatlarini tushuntirib berish –
ijtimoiy pedagogikaning asosiy vazifasi ekanligi barchamizga ma’lum. Ijtimoiy pedagogikaning
asosiy elementlaridan biri bu – ijtimoiylashuv hodisasi hisoblanadi. Ijtimoiylashuv uzoq davom etadigan va juda murakkab jarayon. Bir tomondan har
qanday jamiyat o`z rivojlanish jarayonida ijtimoiy va axloqiy qadriyatlar, ideallar, axloqiy
me’yorlar va qoidalar tizimini ishlab chiqadi, har bir bola jamiyatda mavjud bo‘lgan qonun-
qoidalarni qabul qilib, o`rganib mazkur jamiyatda yashash, uning to`laqonli a’zosi bo`lish
imkoniga ega bo`ladi. Uning uchun jamiyat u yoki bu shaklda shaxsga maqsadga muvofiq tarzda
ta’sir ko`rsatadi, uni ta’lim va tarbiya doirasida amalgi oshiradi. Ikkinchi tomondan, uning
shakllanishiga turli g`oyalar, ijtimoiy muhit ta’sir ko‘rsatadi. Ijtimoiy moslashuv ijtimoiy muhit talablariga insonning munosabat bildirishidir. SHunday qilib, ijtimoiylashuv (moslashuv) individning ijtimoiy mavjudodga aylanish jarayoni
va natijasidir. Indivuallashuv insonning xali yoshligidayoq paydo bo‘ladigan ob’ektiv ehtiyojlari bila
jamiyatdagi o‘z-o‘zini egallashdir. Bu ehtiyoj: j
Indivuallashuv insonning xali yoshligidayoq paydo bo‘ladigan ob’ektiv ehtiyo Indivuallashuv insonning xali yoshligidayoq paydo bo‘ladigan ob’ektiv ehtiyojlari bilan
bog‘liq jamiyatdagi o‘z-o‘zini egallashdir. Bu ehtiyoj: Indivuallashuv insonning xali yoshligidayoq paydo bo ladigan ob ektiv ehtiyojla
og‘liq jamiyatdagi o‘z-o‘zini egallashdir. Bu ehtiyoj: bog‘liq jamiyatdagi o‘z-o‘zini egallashdir. Bu ehtiyoj: a) o‘z qarashlariga ega bo‘lish; a) o‘z qarashlariga ega bo‘lish; b) o‘ziga xosliklariga ega bo‘lish; v) unga tegishli bo‘lgan masalalarni hal qilish, uning o‘z darajasini aniqlab olishiga xalaqit
beradigan hayotiy holatlarga qarshi turish xohishi sifatida namoyon bo‘ladi. v) unga tegishli bo‘lgan masalalarni hal qilish, uning o‘z darajasini aniqlab olishiga xalaqit
beradigan hayotiy holatlarga qarshi turish xohishi sifatida namoyon bo‘ladi. Yuqoridagi fikr-mulohazalardan kelib chiqqan holda, boshlang‘ich sinf o‘quvchilarining
ijtiomiy ko‘nikmalarini shakllantirishga yordam beruvchi bir qancha interfaol usullarni ko‘rib
chiqsak. Eng keng tarqalgan va ancha samarali usul bu – kichik guruhlarga bo‘lib ishlash usulidir. Kichik guruhlarga bo`linadi va ularga reja asosida guruhlarga bo`lib beriladi hamda T- jadval
asosida yoritib berish topshirig`i beriladi. BuxDPI 2-kurs magistrantlari 212
1- guruh
Rahmdillik va xayr-sahovat faoliyati haqida
ijobiy
salbiy
2- guruh
3-guruh
O`zbekistonda xayr-sahovatning qaysi xususiyatlarini guvohi
bo‘lgansiz
kamchiligi
yutuqlari
Rahmdilillik haqida
kamchiligi
yutuqlari 212
|
https://openalex.org/W4232719286
|
https://link.springer.com/content/pdf/10.1007/s12293-018-0260-y.pdf
|
English
| null |
Editorial
|
Memetic computing
| 2,018
|
public-domain
| 1,442
|
Amir H. Alavi1 · Zhihua Cui2 · Amir H. Gandomi1 · Xiao-Zhi Gao3 · Gai-Ge Wang4 · Meng-Hiot Lim5 Published online: 24 May 2018
© Springer-Verlag GmbH Germany, part of Springer Nature 2018 Published online: 24 May 2018
© Springer-Verlag GmbH Germany, part of Springer Nature 2018 Published online: 24 May 2018
© Springer-Verlag GmbH Germany, part of Springer Nature 2018 Recently, inspired by the migration behavior of monarch
butterflies in nature, a metaheuristic optimization algorithm,
called monarch butterfly optimization (MBO), has been pro-
posed. In the present study by Yanhong Feng et al., a novel
chaotic MBO algorithm (CMBO) is developed, in which
chaos theory is introduced in order to enhance its global
optimization ability. Here, a total of 12 one-dimensional
classical chaotic maps are used to tune two main migra-
tion processes of monarch butterflies. Meanwhile, applying
Gaussian mutation operator to some worst individuals can
effectively prevent the premature convergence of the opti-
mization process. The performance of the CMBO is verified
and analyzed by three groups of large-scale 0-1 knapsack
problems instances. The results obtained demonstrate that
the introduction of appropriate chaotic map and Gaussian
perturbation can significantly improve the solution qual-
ity together with the overall performance of the proposed
CMBO algorithm. The CMBO is capable of outperforming
the standard MBO and eight other state-of-the-art canonical
algorithms. Swarm intelligence is a discipline that deals with the nat-
ural and artificial systems composed of many individuals
that coordinate their activities using decentralized control
and self-organization. Lately, swarm intelligence algorithms
are becoming increasingly important in the face of the com-
plexity of today’s demanding applications. This issue aims
to present a collection of recent advances in the swarm
intelligence algorithms. On the basis of a peer-review pro-
cess, six papers are accepted to be included in the thematic
issue, covering various algorithms including particle swarm
optimization, monarch butterfly optimization, moth search
algorithm, evolutionary computation, and cuckoo search. Surrogate assisted meta-heuristic algorithms have recei-
ved increasing attention over the past years, due to the fact
that numerous real-world optimization problems are com-
putationally expensive. However, most existing surrogate
assisted meta-heuristic algorithms are designed for small
or medium scale problems. In the first paper of this issue,
Chaoli Sun et al. propose a fitness approximation assisted
competitive swarm optimizer for the optimization of large
scale expensive problems. Memetic Computing (2018) 10:121–122
https://doi.org/10.1007/s12293-018-0260-y Memetic Computing (2018) 10:121–122
https://doi.org/10.1007/s12293-018-0260-y B Meng-Hiot Lim
EMHLIM@ntu.edu.sg
1
Michigan State University, East Lansing, USA
2
Taiyuan University of Science and Technology, Taiyuan,
China
3
University of Eastern Finland, Kuopio, Finland
4
Ocean University of China, Qingdao, China
5
Nanyang Technological University, Nanyang Avenue,
Singapore Amir H. Alavi1 · Zhihua Cui2 · Amir H. Gandomi1 · Xiao-Zhi Gao3 · Gai-Ge Wang4 · Meng-Hiot Lim5 Different from regular surrogate
assisted evolutionary algorithms that use a computational
model for approximating the fitness, this paper uses esti-
mates of fitness based on the positional relationship between
the individuals in the competitive swarm optimizer. Empir-
ical study on seven widely used benchmark problems with
100 and 500 decision variables show that the proposed fit-
ness approximation assisted competitive swarm optimizer
can achieve competitive performance on a limited compu-
tational budget. The third paper by Gai-Ge Wang proposes a new kind of
metaheuristic algorithm, called moth search (MS) algorithm. In nature, moths are a family of insects associated with butter-
flies belonging to the order Lepidoptera. In the MS method,
the best moth individual is viewed as the light source. Some
moths that are close to the fittest one always display incli-
nation to fly around their own positions in the form of Lévy
flights. On the contrary, due to phototaxis, the moths that
are comparatively far from the fittest one will tend to fly
towards the best one directly in big step. These two features
correspond to the processes of exploitation and exploration
of most metaheuristic optimization methods. The phototaxis
and Lévy flights of the moths can be used to build up a
general-purpose optimization method. In order to demon-
strate the superiority of its performance, the MS method is
further compared with five other up-to-date metaheuristic
optimization algorithms using an array of experiments on 14
basic benchmarks. The results clearly demonstrate that the
MS significantly outperforms five other methods on most of
the test functions and engineering cases. B Meng-Hiot Lim
EMHLIM@ntu.edu.sg
1
Michigan State University, East Lansing, USA
2
Taiyuan University of Science and Technology, Taiyuan,
China
3
University of Eastern Finland, Kuopio, Finland
4
Ocean University of China, Qingdao, China
5
Nanyang Technological University, Nanyang Avenue,
Singapore 12 3 122 Memetic Computing (2018) 10:121–122 problems. Maoqing Zhang et al. present a hybrid multi-
objective CS (HMOCS) for solving multi-objective opti-
mization problems (MOPs). The HMOCS employs the non-
dominated sorting procedure and a dynamical local search. The former is helpful to generate Pareto fronts, and the latter
focuses on enhancing the local search. In order to prove the
capability of the HMOCS, six well-known benchmark MOPs
are used in their experiments. Simulation results show that
HMOCS outperforms three other multi-objective algorithms
in terms of convergence, spread, and distributions. Amir H. Alavi1 · Zhihua Cui2 · Amir H. Gandomi1 · Xiao-Zhi Gao3 · Gai-Ge Wang4 · Meng-Hiot Lim5 The Generalized Vertex Cover Problem (GVCP) extends
classic vertex cover problems to take both vertex and edge
weights into consideration in the objective function. The
GVCP results in finding a vertex subset such that the
sum of vertex weights together with all the correspond-
ing edge weights is minimized. In this paper, Shuli Hu et
al. propose a hybrid metaheuristic algorithm to solve the
GVCP (MAGVCP for short) that is based on evolutionary
search and iterated neighborhood search. The algorithm uses
population initializing procedure to produce high-quality
solutions, applies a dedicated crossover to generate offspring
solutions, and finally utilizes an iterated best chosen neigh-
borhood search to find better solutions. Experiments carried
on random instances and DIMACS instances verify the effec-
tiveness of the proposed algorithm. It has indeed been a pleasure to edit this special issue. We are grateful to all the authors who submitted their papers
and to the reviewers for their timely and insightful reviews
that have greatly helped improve the quality of this issue. We
hope that the readers find the papers collected in this thematic
issue interesting and helpful. The Guest Editors acknowledge
the financial supports from the Natural Science Foundation
of Shanxi Province under Grant No. 201601D011045. The fifth paper by Heqi Wang et al. presents a robust opti-
mization algorithm based on the hybridization of the krill
herd (KH) and artificial bee colony (ABC) methods and
the information exchange concept. The global optimal solu-
tions found by the proposed hybrid KH and ABC (KHABC)
algorithm are considered as a neighbor food source for
the onlooker bees in the ABC. Thereafter, a local search
is performed by the onlooker bees in order to find a bet-
ter solution around the given neighbor food source. Both
methods, the KH and ABC, share the globally best solu-
tions through the information exchange process between the
krill and bees. Based on the above results, the exchange pro-
cess significantly improves the exploration and exploitation
of the hybrid method. Besides, a focused elitism scheme
is introduced to enhance the performance of the developed
algorithm. The validity of the KHABC method is validated
using 13 unconstrained benchmark functions, a total of 21
CEC 2017 constrained real-parameter optimization prob-
lems, and ten CEC 2011 real world problems (RWPs). The
proposedmethodclearlydemonstratesitspotentialforacom-
petitive optimization tool towards solving both benchmark
functions and real world problems. Notes from Editor-in-Chief f
f
The above six articles make up the Thematic Issue on Novel
Swarm Intelligence Algorithms. To further complement this
thematic issue, we include two more regular issue papers
on applications to complete the issue. The paper by Nodeh
et. al. describes a hybrid algorithm that combines differen-
tial evolution and mutable smart bee algorithm. The work
has practical significance, being applied to optimize the fab-
rication of piezoelectric excited millimetre-sized cantilever
sensor. Such a problem usually involves constraints, and the
authors show that their approach can optimize the design
parameters for the sensors. Another paper by Liu et al. deals
with active object recognition based on sequence of actions. This is achieved by means of camera with adjustable posi-
tion and orientation. They use extreme learning machine to
learn the state representations and the strategy for reinforce-
ment learning, which is validated based on a public-domain
dataset. Cuckoosearch(CS)isarecentlydevelopedmeta-heuristic,
which has shown good search abilities on many optimization Publisher’s Note Springer Nature remains neutral with regard to juris-
dictional claims in published maps and institutional affiliations. 123 123
|
https://openalex.org/W2963614914
|
https://mathematicsinindustry.springeropen.com/track/pdf/10.1186/s13362-018-0057-0
|
English
| null |
On impedance conditions for circular multiperforated acoustic liners
|
Journal of mathematics in industry
| 2,018
|
cc-by
| 11,569
|
R ES EA RCH Open Access © The Author(s) 2018. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License
(http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, pro-
vided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and
indicate if changes were made. *Correspondence:
kschmidt@mathematik.tu-darmstadt.de
1Fachbereich Mathematik, AG
Numerik und Wissenschaftliches
Rechnen, Technische Universität
Darmstadt, Darmstadt, Germany
Full list of author information is
available at the end of the article Kersten Schmidt1*
, Adrien Semin1, Anastasia Thöns-Zueva2 and Friedrich Bake3 Kersten Schmidt1*
, Adrien Semin1, Anastasia Thöns-Zueva2 and Friedrich Bake3 *Correspondence:
kschmidt@mathematik.tu-darmstadt.de
1Fachbereich Mathematik, AG
Numerik und Wissenschaftliches
Rechnen, Technische Universität
Darmstadt, Darmstadt, Germany
Full list of author information is
available at the end of the article Abstract Background: The acoustic damping in gas turbines and aero-engines relies to a great
extent on acoustic liners that consists of a cavity and a perforated face sheet. The
prediction of the impedance of the liners by direct numerical simulation is nowadays
not feasible due to the hundreds to thousands repetitions of tiny holes. We introduce
a procedure to numerically obtain the Rayleigh conductivity for acoustic liners for
viscous gases at rest, and with it define the acoustic impedance of the perforated
sheet. Results: The proposed method decouples the effects that are dominant on different
scales: (a) viscous and incompressible flow at the scale of one hole, (b) inviscid and
incompressible flow at the scale of the hole pattern, and (c) inviscid and compressible
flow at the scale of the wave-length. With the method of matched asymptotic
expansions we couple the different scales and eventually obtain effective impedance
conditions on the macroscopic scale. For this the effective Rayleigh conductivity
results by numerical solution of an instationary Stokes problem in frequency domain
around one hole with prescribed pressure at infinite distance to the aperture. It
depends on hole shape, frequency, mean density and viscosity divided by the area of
the periodicity cell. This enables us to estimate dissipation losses and transmission
properties, that we compare with acoustic measurements in a duct acoustic test rig
with a circular cross-section by the German Aerospace Center in Berlin. Conclusions: A precise and reasonable definition of an effective Rayleigh
conductivity at the scale of one hole is proposed and impedance conditions for the
macroscopic pressure or velocity are derived in a systematic procedure. The
comparison with experiments show that the derived impedance conditions give a
good prediction of the dissipation losses. MSC: 35Q30; 35B27; 74Q15; 76M50 Keywords: Acoustic liner; Perforated plates; Multiscale analysis; Rayleigh
conductivity; Impedance conditions Keywords: Acoustic liner; Perforated plates; Multiscale analysis; Rayleigh
conductivity; Impedance conditions ( 2018) 8:15 ( 2018) 8:15 Schmidt et al. Journal of Mathematics in Industry
https://doi.org/10.1186/s13362-018-0057-0 1 Introduction The safe and stable operation of modern low-emission gas turbines and aero-engines cru-
cially depends on the acoustic damping capability of the combustion system components. The safe and stable operation of modern low-emission gas turbines and aero-engines cru-
cially depends on the acoustic damping capability of the combustion system components. Hereby, so called bias flow liners—consisting of a cavity and a perforated face sheet with
additional cooling air flow—play a significant role. Since decades the damping perfor-
mance prediction of these bias flow liners under all possible flow conditions remains a ( 2018) 8:15 Page 2 of 20 Schmidt et al. Journal of Mathematics in Industry major challenge. However, due to the higher tendency of low-emission, lean burn com-
bustion concepts for combustion instabilities the prediction of the acoustic bias flow liner
impedance and therewith its damping performance is a very important prerequisite for the
engine design process. Several analytical and semi-empirical models for the impedance
description of bias flow liners were developed in the past (see also [1]). This work focuses
on the numerical simulation of the acoustic characteristics of bias flow liners applying
multi-scale modeling. In principal all theoretical approaches are based on the formulation of the Rayleigh con-
ductivity KR [2, 3], which describes the ratio of the fluctuating volume flow Q(t) through
a hole to the driving pressure difference P–(t) – P+(t) across the hole: KR :=
ρ0∂tQ(t)
P–(t) – P+(t),
(1) KR :=
ρ0∂tQ(t)
P–(t) – P+(t), (1) and has the dimensions of length. One major challenge in the model description of the
Rayleigh conductivity represents the definition or the specification of the pressure differ-
ence since, above and below the perforated liner face-sheet the pressure is not necessarily
constant rather a function of the distance from the hole. Here, the present work applying
a multiscale asymptotic model will provide an exactly defined solution. More precisely,
the Rayleigh conductivity of a single hole in an array of holes is distributed over the whole
liner area. In this way the effective Rayleigh conductivity kR = KR
Aδ
(2) kR = KR
Aδ (2) as quotient of the Rayleigh conductivity of one hole and the area Aδ of one periodicity cell
of the array is introduced that has the dimensions of one over length. Using the effective
Rayleigh conductivity the liner impedance can be determined like later shown for example
in Eq. (21). 1 Introduction Indeed, the (effective) Rayleigh conductivity depends on the geometrical parameters,
especially size and shape of the holes and their distances as well as the physical parameters,
noting at first the viscosity and the frequency. Some of these parameters take small values
and we consider them to be scaled with a small parameter δ > 0 in such a way that making
δ smaller the effective Rayleigh conductivity stays essentially constant and tends in the
limit δ →0 to a non-trivial value. 2 Methods We consider an acoustic liner that consist of a wall or part of a wall with a periodic dense
array of equisized and equishaped holes with a characteristic periodicity that is propor-
tional to the small parameter δ. The holes may not be of cylindrical shape and even tilted
in general. For sake of simplicity we consider the perforated wall Ωδ
liner with a circular
cross-section of fixed inner radius Rd, while noting that the proposed procedure to define
the Rayleigh conductivity and impedance conditions do not depend on the choice of the
cross-section, but only on the hole pattern and hole shape and can be directly transfered
to other cross-sections like rectangular. To derive the impedance conditions we let the parameter δ of the hole period tending to
zero—so the number of holes increases accordingly—while the inner and outer diameter ( 2018) 8:15
Page 3 of 20 Schmidt et al. Journal of Mathematics in Industry ( 2018) 8:15
Page 3 of 20 Schmidt et al. Journal of Mathematics in Industry ( 2018) 8:15 ( 2018) 8:15 Page 3 of 20 Page 3 of 20 Schmidt et al. Journal of Mathematics in Industry Figure 1 (a) Simplified geometry of a combustion liner for acoustic studies. (b) Flattened liner. (c) View along
a cross-section Figure 1 (a) Simplified geometry of a combustion liner for acoustic studies. (b) Flattened liner. (c) View along
a cross-section Figure 2 Split of the domain Ω into the two semi-infinite waveguides Ω± and the multi-perforated liner
section Ωc. Impedance transmission conditions on the interface Γliner approximate the behaviour of the
many perforations Figure 2 Split of the domain Ω into the two semi-infinite waveguides Ω± and the multi-perforated liner
section Ωc. Impedance transmission conditions on the interface Γliner approximate the behaviour of the
many perforations of the cross section are scaled like δ2 as well as the thickness of the perforated wall, see
Fig. 1. As δ →0, the holes merge and the domain Ωδ
liner degenerates to an interface Γliner,
on which we will prescribe the impedance conditions representing the correct dissipation
behaviour of the acoustic liner (see Fig. 2). For the circular liner the limit interface domain
Γliner is a cylinder of radius Rd. 2 Methods As it simplifies the derivation and resulting formula of
the impedance conditions greatly we assume for the area Aδ of the periodicity cell of the
periodic array that Aδ = δ2. This liner shall be embedded in a duct domain Ω and the computational domain is
Ωδ := Ω \ Ωδ
liner for every δ > 0, i.e., the duct domain without the multi-perforated wall. On this domain we introduce as viscoacoustic model the linearized compressible Navier-
Stokes equations in frequency domain in a uniform and stagnant media for a source term
f(t,x) = Re(f(x)exp(–ıωt)) with an angular frequency ω > 0: –ıωvδ + 1
ρ0
∇pδ – ν(δ) vδ – ν′(δ)∇divvδ = f,
in Ωδ,
(3a)
–ıωpδ + ρ0c2 divvδ = 0,
in Ωδ,
(3b)
vδ = 0,
on ∂Ωδ,
(3c) (3a) (3c) with the acoustic velocity vδ, the acoustic pressure pδ, the mean density ρ0 > 0, the speed
of sound c, the kinematic and secondary viscosities ν(δ),ν′(δ) > 0. We scale the viscosities
for δ →0 like δ4 such that the size of the viscous boundary layers remain asymptotically
the same at the scale of a single hole. If the duct is modelled to be of infinite extend then
additional conditions at infinity have to be posed. Assuming a channel of constant cross-
section and no incoming wave from infinity it suffices that the pressure decays at infinity, with the acoustic velocity vδ, the acoustic pressure pδ, the mean density ρ0 > 0, the speed
of sound c, the kinematic and secondary viscosities ν(δ),ν′(δ) > 0. We scale the viscosities
for δ →0 like δ4 such that the size of the viscous boundary layers remain asymptotically
the same at the scale of a single hole. If the duct is modelled to be of infinite extend then
additional conditions at infinity have to be posed. Assuming a channel of constant cross-
section and no incoming wave from infinity it suffices that the pressure decays at infinity, ( 2018) 8:15
Page 4 of 20 hematics in Industry ( 2018) 8:15
Page 4 of ( 2018) 8:15 Page 4 of 20 Schmidt et al. 2 Methods Journal of Mathematics in Industry i.e., limz→±∞pδ = 0, which is equivalent to pose Dirichlet-to-Neumann boundary condi-
tions at z = ±R i.e., limz→±∞pδ = 0, which is equivalent to pose Dirichlet-to-Neumann boundary condi-
tions at z = ±R
pδ
√ν curlvδ × n
(±R) – Λ±(ν)
vδ · n
vδ × n
(±R) = 0
(3d) (3d) with n = ±(1,0,0)⊤and any R > 0 large enough, where Λ± can be derived in analogy to 2D
wave-guides [4] based on outgoing guided and evanescent modes. Note, that this condi-
tion is stated only for completeness. As it is independent of the presence of the acoustic
liner the derivation will not be detailed. Moreover, for an incoming wave from infinity
we can always use a reformulation with a source term f, and we assume this source to be
located away from the perforated wall such that f = 0 in a neighbourhood. In the following section we study the solution of the viscoacoustic model in three dif-
ferent geometrical scales beginning at the scale of one hole, pursuing with the scale of
one period of the hole array and concluding with the macroscopic scale, on which the
impedance conditions follow. 2.1 Microscopic scale: the near field around one hole Journal of Mathematics in Industry ( 2018) 8:15 ( 2018) 8:15 Page 5 of 20 Schmidt et al. Journal of Mathematics in Industry where
Ωhole is the scaled domain representing one hole, and we assume 0 ∈
Ωhole. For in-
stance a vertical cylindrical hole of diameter d0δ2 can be represented by
Ωhole = {(r,y,z) ∈
R3 such that 0 ≤r ≤h0 and y2 + z2 < 1
4d2
0}. where
Ωhole is the scaled domain representing one hole, and we assume 0 ∈
Ωhole. For in-
stance a vertical cylindrical hole of diameter d0δ2 can be represented by
Ωhole = {(r,y,z) ∈
R3 such that 0 ≤r ≤h0 and y2 + z2 < 1
4d2
0}. Close to one hole of the perforated liner, we represent the solution (vδ,pδ) of (3a)–(3d)
as as as vδ = δ–2v–2(xΓ ,X) + O
δ–1
,
pδ = p0(xΓ ,X) + O(δ),
(5) (5) where the near field corrector terms (v–2,p0) do not depend on δ. where the near field corrector terms (v–2,p0) do not depend on δ. Now, inserting expansion (5) into the viscoacoustic model (3a)–(3d) and identifying for-
mally terms of same powers of δ results in a product representation of the near-field cor-
rector
v–2(xΓ ,X),p0(xΓ ,X)
= c(xΓ )
˜v(X), ˜p(X)
, where c(xΓ ) allows for a slow variation of near field velocity and pressure along the wall. The near field profiles (˜v, ˜p) are solution of the instationary Stokes problem where c(xΓ ) allows for a slow variation of near field velocity and pressure along the wall. The near field profiles (˜v, ˜p) are solution of the instationary Stokes problem –ıω ˜v + 1
ρ0
∇˜p – ν0 ˜v = 0,
in
Ω,
(6a)
div ˜v = 0,
in
Ω,
(6b)
˜v = 0,
on ∂
Ω,
(6c) (6a) div ˜v = 0,
in
Ω, (6b) (6c) where ∇, div and are the gradient, divergence and Laplace operator in X (cf. [5,
Sect. 2.1.6] in time-domain). The near field velocity profile is incompressible on the scale
of one hole and fulfills together with the near field pressure profile the Stokes equations
with a significant viscosity ν0 at the scale of one hole and the additional term –iω ˜v that
reflects a time shift between excitation and excited fields. 2.1 Microscopic scale: the near field around one hole In the vicinity of one hole that tends to a point xΓ on the interface Γliner we use the local
coordinate X := (r,y,z) = ((r – Rd)/δ2,rθ/δ2,z/δ2). As δ →0, the hole variable X occupies
the whole unbounded domain
Ω defined by (see Fig. 3(a)) In the vicinity of one hole that tends to a point xΓ on the interface Γliner we use the local
coordinate X := (r,y,z) = ((r – Rd)/δ2,rθ/δ2,z/δ2). As δ →0, the hole variable X occupies
the whole unbounded domain
Ω defined by (see Fig. 3(a))
Ω =
(r,y,z) ∈R3 such that r < 0 or r > h0
∪
Ωhole,
(4)
Ω =
(r,y,z) ∈R3 such that r < 0 or r > h0
∪
Ωhole,
(4) (4) Figure 3 (a) Computational domain for the near field problem around a single hole. (b) The near field
pressure (real part) for the liner configuration DC006 (see Table 1) at ω = 2π × 306 s–1 using S = 40. (c), (d) The near field velocity (imaginary part) for the same configuration as (b) and for ω = 2π × 30 s–1. Here, the color coding corresponds to the amplitude and the arrows to the direction of the velocity Figure 3 (a) Computational domain for the near field problem around a single hole. (b) The near field
pressure (real part) for the liner configuration DC006 (see Table 1) at ω = 2π × 306 s–1 using S = 40. (c), (d) The near field velocity (imaginary part) for the same configuration as (b) and for ω = 2π × 30 s–1. Here, the color coding corresponds to the amplitude and the arrows to the direction of the velocity Figure 3 (a) Computational domain for the near field problem around a single hole. (b) The near field
pressure (real part) for the liner configuration DC006 (see Table 1) at ω = 2π × 306 s–1 using S = 40. (c), (d) The near field velocity (imaginary part) for the same configuration as (b) and for ω = 2π × 30 s–1. Here, the color coding corresponds to the amplitude and the arrows to the direction of the velocity Schmidt et al. Journal of Mathematics in Industry ( 2018) 8:15
Page 5 ( 2018) 8:15
Page 5 of 20 Schmidt et al. 2.1 Microscopic scale: the near field around one hole These equations are completed
by Dirichlet jump conditions at infinity where ∇, div and are the gradient, divergence and Laplace operator in X (cf. [5,
Sect. 2.1.6] in time-domain). The near field velocity profile is incompressible on the scale
of one hole and fulfills together with the near field pressure profile the Stokes equations
with a significant viscosity ν0 at the scale of one hole and the additional term –iω ˜v that
reflects a time shift between excitation and excited fields. These equations are completed
by Dirichlet jump conditions at infinity lim
S→∞˜p|Γ±(S) = ±1
2,
(6d) lim
S→∞˜p|Γ±(S) = ±1
2, (6d) that act as a excitation from far away and will be used for the matching with the mesoscopic
scale (see Sect. 2.2). Here, Γ±(S) =
(r,y,z) ∈
Ω,±r > r± and (r – r±)2 + y2 + z2 = S2
,
(7) (7) with r– = 0 and r+ = h0, are the two half-spheres (see Fig. 3(a)) that are moved towards
infinity. with r– = 0 and r+ = h0, are the two half-spheres (see Fig. 3(a)) that are moved towards
infinity. Note that in problem (6b)–(6d) the term –ν′
0∇div ˜v that would appear in the first line
cancels out due to the divergence free condition (6b). Moreover, note that the term –ν0 ˜v
can be replaced by ν0 curlcurl ˜v and so only the vorticity part of the velocity ˜v will exhibit
a viscosity boundary layer as we will see later. Problem (6b)–(6d) is a classical saddle-point problem and admits a unique solution
stated by the following proposition. ( 2018) 8:15
Page 6 of 20 ( 2018) 8:15 Page 6 of 20 Schmidt et al. Journal of Mathematics in Industry Proposition 2.1 There exists a unique solution (˜v, ˜p) ∈(H1(
Ω))3 × V(
Ω) of (6b)–(6d),
where V(
Ω) = {P ∈H1
loc(
Ω) such that ∥∇P∥L2(
Ω) < ∞}. Note, that the pressure space V(
Ω) allows for a constant behavior towards infinity. Note, that the pressure space V(Ω) allows for a constant behavior towards infinity. 2.1 Microscopic scale: the near field around one hole With the near field velocity profile ˜v defined by (6b)–(6d) we can define in analogy to
the Rayleigh conductivity a posteriori the quantity kR := lim
S→∞
ıωρ0
2
Γ+(S)
˜v · n –
Γ–(S)
˜v · n
(8) (8) using the volume flux towards infinity in a symmetric way. Here, n is the outer normal
vector. In this way, the quantity kR is a mapping of a constant near field pressure at infinity
to the flux at infinity. To see the analogy it suffices to consider time harmonic fields vary-
ing like exp(–ıωt), the volume flux Q(t) through the aperture counted positively along the
direction of the er axis to be the same as the volume flux through the surface Γ+(S) (re-
spectively Γ–(S)), counted positively (resp. negatively) along the direction of the normal
vector n, and to compare (1) and (8). Note, that the normal component of the near field velocity profile v decays like 1/S2
towards infinity and combines different behaviour close to and away from the wall (see
Fig. 3(c) and (d)). This behaviour can be rigorously justified with similar techniques as in
[6, 7]. For the usual definition of the Rayleigh conductivity KR it is not evident where the differ-
ence of the pressure—as it varies locally—and the volume flux—as in the original acous-
tic equations the fluid is compressible—shall be evaluated. The quantity kR is, however,
clearly defined by (6b)–(6d) and (8) as the near field pressure tends to constant values for
|X| →∞and as the near field velocity is incompressible. This results from the separation
of the effects at the different length scales, namely viscous incompressible behaviour in
the vicinity of the holes versus inviscid, compressible behaviour away from them, due to
the asymptotic ansatz. As the near field profiles are defined in local coordinates X it has
the dimensions of one over length and we denote it as effective Rayleigh conductivity of
the liner. The definition of the effective Rayleigh conductivity kR can be used for inviscid fluids as
well, for which ν0 = 0, if the no-slip boundary conditions (6c) are replaced by v · n = 0. 2.2 Mesoscopic scale: the hole pattern Pursuing with the scale of one period of the hole array and in the vicinity of one hole that
tends to xΓ , we use the local coordinate X := (R,Y,Z) = ((r – Rd)/δ,rθ/δ,z/δ). We consider
for fixed δ > 0 the infinite periodicity cell (see Fig. 4) Bδ = Bδ
+ ∪Bδ
– ∪δ
Ωhole,
(9) (9) Bδ = Bδ
+ ∪Bδ
– ∪δ
Ωhole, where Bδ
± = {(R,Y,Z) ∈R3 such that |Y – bZ| <
√a
2 ,|Z| <
1
2√a,±R > Rδ
±} with Rδ
– = 0,
Rδ
+ = h0δ are two semi-infinite parallelepipeds whose opposite lateral faces |Z| =
1
2√a and
|Y – bZ| =
√a
2 are considered to be identified with each other such that Bδ
± and so Bδ are
topologically equivalent to a torus. With the cross-section of the periodicity cell Γ (S) =
(R,Y,Z) ∈R3 such that |Y – bZ| <
√a
2 ,|Z| <
1
2√a,R = S Schmidt et al. Journal of Mathematics in Industry ( 2018) 8:15
Page 7 of 20 ( 2018) 8:15 Page 7 of 20 Schmidt et al. Journal of Mathematics in Industry Figure 4 Representation of the periodicity cell Bδ associated to the intermediate scale Figure 4 Representation of the periodicity cell Bδ associated to the intermediate scale the symmetric difference A△B := (A ∪B) \ (A ∩B) the boundary of the periodicity cell is
given as ∂Bδ = (Γ (h0δ) ∪Γ (0))△δ∂
Ωhole. It consists of the wall boundary and the bound-
ary of the hole. The periodicity cell Bδ degenerates as δ →0 and tends to the union B0 of
two semi-infinite parallelepipeds B0
± connected by the point 0, an infinitely small hole. I
id th
i di
f h l
t th
l ti
( δ
δ) f (3 ) (3d) vδ = Vδ
0(xΓ ,X) + O(δ),
pδ = Pδ
0(xΓ ,X) + δPδ
1(xΓ ,X) + O
δ2
(10) vδ = Vδ
0(xΓ ,X) + O(δ), (10) pδ = Pδ
0(xΓ ,X) + δPδ
1(xΓ ,X) + O
δ2 with X ∈Bδ. Inserting expansion (10) in problem (3a)–(3d) and identifying formally the terms of
same powers of δ gives that Pδ
0(xΓ ,X) is constant in X and a separation of variables for
the mesoscopic corrector as (Vδ
0(xΓ ,X),Pδ
1(xΓ ,X)) = c(xΓ )(Vδ(X),Pδ(X)) with the meso-
scopic profile (Vδ,Pδ) satisfying the Darcy-type problem ⎧
⎪⎪⎨
⎪⎪⎩
–ıωVδ + 1
ρ0 ∇Pδ = 0,
in Bδ,
divVδ = 0,
in Bδ,divVδ = 0,
in Bδ,
Vδ · n = 0,
on ∂Bδ. (11) (11) Here, ∇and div are the gradient and divergence in X. The formal identification of terms
of same power in δ can be justified despite the fact that the size of the hole depends on δ
as well. For this an additional scale η for the size of one hole has to be introduced that is
first considered to be independent of δ due to its different meaning and later fixed to δ2. The expansion (10) is then in δ, where the terms of the expansion depend on η. For the
brevity of the article we have chosen directly η = δ2. Note that (11) is equivalent to an homogeneous Laplace problem with Neumann bound-
ary conditions for the pressure profile Pδ, where the velocity profile Vδ can be computed
from. Following [8, Proposition 2.2], we can therefore state the following proposition. Proposition 2.2 For any fixed δ > 0, the kernel of problem (11) is of dimension 2 and
spanned by the functions (Vδ
N ,Pδ
N ) = (0,1) and (Vδ
D,Pδ
D) such that Vδ
D tends to a con-
stant as R →±∞. Moreover, there exists Dδ
∞∈C such that we can choose (Vδ
D,Pδ
D) with Schmidt et al. Journal of Mathematics in Industry ( 2018) 8:15 ( 2018) 8:15 Page 8 of 20 Schmidt et al. Journal of Mathematics in Industry the limit behaviour: the limit behaviour: Vδ
D = 1
ıωeR + o(1),
R →±∞,
Pδ
D = ρ0R ± Dδ
∞+ o(1),
R →±∞. (12) Vδ
D = 1
ıωeR + o(1),
R →±∞,
Pδ
D = ρ0R ± Dδ
∞+ o(1),
R →±∞. (12) It remains to determine the constant Dδ
∞, where we are in particular interested in its
asymptotic behaviour for δ →0. with X ∈Bδ. To obtain this behaviour we will match the mesoscopic
functions Vδ
D and Pδ
D with the near field profiles ˜v and ˜p at half-spheres Γ±(sδ) of radius sδ
for
√
δ < sδ < 2
√
δ centered at the aperture 0. First we note that due to the incompressibility
and the limit behaviour of Vδ
D for its volume flux over the half-spheres it holds ıω
2
Γ+(sδ)
Vδ
D · n –
Γ–(sδ)
Vδ
D · n
= ıω
2 lim
S→∞
Γ (S)
Vδ
D · eR +
Γ (–S)
Vδ
D · eR = 1. Using this equality, definition (8) of the effective Rayleigh conductivity kR, the mesoscopic
to microscopic variable change X = X/δ, and matching of the mesoscopic velocity Vδ
D and
the near field velocity profile ˜v we find that Vδ
D(X) ∼ρ0
kRδ2 ˜v
X
δ
for
√
δ < |X| < 2
√
δ and δ →0, and this relation explains a posteriori the expansion (5). By linearity and using defini-
tion of problems (6b)–(6d) and (11), the gradient of the mesoscopic pressure Pδ
D can be
matched with the gradient of the near field pressure profile as well. Integrating these gra-
dients, using limit (6d) and Proposition 2.2 leads to Pδ
D(X) ∼ρ0
kRδ ˜p
X
δ
∼± ρ0
2kRδ
for
√
δ < |X| < 2
√
δ,±R > 0 and δ →0. As for δ →0 the mesoscopic pressure Pδ
D tends to ρ0R ± Dδ
∞if δ = o(|X|) we conclude
that Dδ
∞=
ρ0
2kRδ + o
δ–1
. (13) Dδ
∞=
ρ0
2kRδ + o
δ–1
. (13) This blow up of the coefficient Dδ
∞as δ →0 in accordance with its numerical computa-
tions based on an asymptotic analysis of (3a)–(3d) with only two scales [9], where the hole
size is considered not as a scale but as a parameter. This blow up of the coefficient Dδ
∞as δ →0 in accordance with its numerical computa-
tions based on an asymptotic analysis of (3a)–(3d) with only two scales [9], where the hole
size is considered not as a scale but as a parameter. 2.3 Macroscopic scale and impedance conditions Finally, away from a vicinity of the layer, the solution (vδ,pδ) of (3a)–(3d) is represented
by vδ(x) = v0(x) + o(1),
pδ(x) = p0(x) + o(1). (14) vδ(x) = v0(x) + o(1),
pδ(x) = p0(x) + o(1). vδ(x) = v0(x) + o(1), (14) pδ(x) = p0(x) + o(1). ( 2018) 8:15
Page 9 of 20 Schmidt et al. Journal of Mathematics in Industry ( 2018) 8:15
Page 9 of 20 ( 2018) 8:15 Schmidt et al. Journal of Mathematics in Industry Page 9 of 20 Inserting this expansion in problem (3a)–(3d) and making a formal identification in terms
of powers of δ gives that (v0,p0) is solution of the classical Helmholtz problem –ıωv0 + 1
ρ0
∇p0 = f,
in Ω \ Γliner,
(15a) –ıωv0 + 1
ρ0
∇p0 = f,
in Ω \ Γliner,
(15a)
–ıωp0 + ρ0c2 divv0 = 0,
in Ω \ Γliner,
(15b) (15a) –ıωp0 + ρ0c2 divv0 = 0,
in Ω \ Γliner,
(15b) (15b) –ıωp0 + ρ0c2 divv0 = 0,
in Ω \ Γliner,
(15b) and a multiscale analysis, for that some theory can also be found in [10] for rigid walls
leads, to the boundary conditions and a multiscale analysis, for that some theory can also be found in [10] for rigid walls
leads, to the boundary conditions v0 · n = 0,
on ∂Ω. (15c) (15c) v0 · n = 0,
on ∂Ω. v0 · n = 0,
on ∂Ω. In analogy to the 2D case [9] it can be shown that the Dirichlet-to-Neumann condition
(3d) degenerates as well and becomes p0(±R) – Λ±(0)v0 · n(±R) = 0,
(15d) p0(±R) – Λ±(0)v0 · n(±R) = 0,
(1 (15d) where the definition of Λ±(0) is based on outgoing guided and evanescent modes (see [11,
Eq. (2.7)]). This problem is completed by jump conditions across the interface Γliner. To obtain the
conditions we match the macroscopic pressure p0 and flux v0 · n in a matching zone at
distance
√
δ to the interface Γliner to the mesoscopic pressure and velocity functions. 2.3 Macroscopic scale and impedance conditions 1), but only on their area Aδ, namely through
ν0 = ν/A2
δ in the computation of the effective Rayleigh conductivity kR. Distinguished limit
Note, that the nature of the impedance condition (19b) is due to
the choice of asymptotic scales. It represents a distinguished limit meaning that different
choice would lead to one of the trivial conditions [p0](xΓ ) = 0 (transparent wall) or ⟨v0 ·
n⟩(xΓ ) = 0 (rigid wall), as it was already stated in [12, Eq. (4.4) and below] for infinitely thin
perforated wall and the Stokes flows. If we would scale the diameter of each hole with ε(δ)
as well as the thickness of the perforated wall such that δ2 = o(ε(δ)) then we would obtain
transparent wall conditions in the limit δ →0, e.g., if the diameter of each hole scales like δ. A contrario, the impedance conditions become rigid wall conditions if we would use the
scaling ε(δ) = o(δ2). Acoustic impedance
The nature of the impedance conditions is known in the literature:
the notion of impedance can be found in the works of Webster in the 1910s [13]. More
precisely, he defines the normalized specified acoustic impedance ζ by (note there is a
complex conjugate and a different sign due to the different choice of the time-dependency
convention) ζ := –
[p0]
cρ0⟨v0 · n⟩
. (20) (20) For the derived impedance conditions (19b) and by identification, the normalized speci-
fied acoustic impedance for perforated walls is given by For the derived impedance conditions (19b) and by identification, the normalized speci-
fied acoustic impedance for perforated walls is given by ζ = ıω
ckR
= ıωkR
c|kR|2 . (21) ζ = ıω
ckR
= ıωkR
c|kR|2 . (21) The resistance Re(ζ) and the reactance Im(ζ) are positive quantities when kR has a positive
real part and a negative imaginary part. Moreover in the inviscid case kR is a positive real
number, so that the normalized specified acoustic impedance ζ is purely a reactance. 2.3 Macroscopic scale and impedance conditions For
the pressure we find p0(x) = CN (xΓ )Pδ
N
x – xΓ
δ
+ δCD(xΓ )Pδ
D
x – xΓ
δ
for
√
δ ≤|x – xΓ | ≤2
√
δ and δ →0,
(16) (16) or
√
δ ≤|x – xΓ | ≤2
√
δ and δ →0,
(16) for
√
δ ≤|x – xΓ | ≤2
√
δ and δ →0, with two functions CN , CD that allow for slow variation along the perforated wall. With
the factor δ the limit δPδ
D( x–xΓ
δ
) for δ →0 becomes ± ρ0
2kR for ±(x – xΓ ) · n > 0, using (13). Subtracting the two limits of (16) for δ →0 we obtain [p0](xΓ ) := lim
δ→0p0(xΓ +
√
δn) – p0(xΓ –
√
δn) = CD(xΓ )ρ0
kR
. (17) (17) Taking the gradient in x on both sides of (16) and using (15a), the assumption that f = 0
close to the perforated wall and (11) we find v0(x) · n = ρ0
ıω∇p0(x) · n = δCD(xΓ ) ρ0
ıω∇Pδ
D
x – xΓ
δ
= CD(xΓ )Vδ
D
x – xΓ
δ
for
√
δ ≤|x – xΓ | ≤2
√
δ and δ →0. (18) (18) As the two limits for Vδ
D for R →±∞coincide by Proposition 2.2 we obtain As the two limits for Vδ
D for R →±∞coincide by Proposition 2.2 we obtain [v0 · n](xΓ ) = 0,
on Γliner. (19a) [v0 · n](xΓ ) = 0,
on Γliner. (19a) ( 2018) 8:15
Page 10 of 20 ( 2018) 8:15 Page 10 of 20 Schmidt et al. Journal of Mathematics in Industry Finally, taking the average of (18), using that limR→±∞Vδ
D =
1
ıωeR (see Proposition 2.2)
and taking the limit δ →0 gives in view of (17) the impedance conditions [p0](xΓ ) = ıωρ0
kR
⟨v0 · n⟩(xΓ ),
on Γliner. (19b) (19b) Note, that the impedance conditions do not depend on the pattern of the holes, more
precisely on the values a and b (see Fig. 1), but only on their area Aδ, namely through
ν0 = ν/A2
δ in the computation of the effective Rayleigh conductivity kR. Note, that the impedance conditions do not depend on the pattern of the holes, more
precisely on the values a and b (see Fig. 3 Results and discussion In this section, we are interested by the numerical computation of the effective Rayleigh
conductivity kR, the computation of dissipation losses in acoustic ducts with the imped-
ance conditions and comparison with data from experimental measurements. 2.3 Macroscopic scale and impedance conditions Formulation in pressure only
One can also remark that Problem (15a)–(15d) can be for-
mulated in terms of pressure only: equations (15a)–(15d) give p0 + ω2
c2 p0 = divf,
in Ω \ Γliner, p0 + ω2
c2 p0 = divf,
in Ω \ Γliner, ∇p0 · n = 0,
on ∂Ω,
(22a) (22a) ∇p0 · n = 0,
on ∂Ω, lim
z→±∞±∂zp0 – ıωρ0T1
±p0 = 0, lim
z→±∞±∂zp0 – ıωρ0T1
±p0 = 0, ( 2018) 8:15
Page 11 of 20 ( 2018) 8:15 Page 11 of 20 Schmidt et al. Journal of Mathematics in Industry and impedance conditions (19a)–(19b) are written in terms of the pressure as
[∇p0 · n](xΓ ) = 0
and
⟨∇p0 · n⟩(xΓ ) = kR[p0](xΓ ). (22b and impedance conditions (19a)–(19b) are written in terms of the pressure as (22b) [∇p0 · n](xΓ ) = 0
and
⟨∇p0 · n⟩(xΓ ) = kR[p0](xΓ ). (22b) This study was done for the inviscid case [14, 15], for which kR is called effective plate com-
pliance. In these papers, the authors derived an equivalent porous condition [15, Eq. (16)]
and effective reflection and transmission coefficients [15, Eq. (5.17–19)], the difference is
due to the choice of the scale for the geometry. 3.1 Numerical computation of kR The effective Rayleigh conductivity kR is defined through the solution of the near field ve-
locity and pressure profiles in the unbounded domain
Ω around a single hole. To compute
kR numerically we truncate the unbounded domain, on which we use the finite element
method for discretization and propose an extrapolation procedure to increase the accu-
racy. First, we define the truncated domain First, we define the truncated domain
Ω(S) =
Ω ∩
max
X – (0,0,0)
,
X – (h0,0,0)
< S
(23) (23) of
Ω for a given truncation radius S > 0.5
d2
0 + h2
0 (see Fig. 3(a)). It has two artificial
boundaries Γ±(S) that are no boundaries of
Ω. We restrict the problem (6b)–(6d) to
Ω(S)
and ∂
Ω(S) ∩∂
Ω, and we approximate the conditions (6d) by setting ˜p|Γ±(S) = ±1
2. ˜p|Γ±(S) = ±1
2. From the resolution of the truncated problem we compute the approximated Rayleigh
conductivity kR(S) taking as well an approximation of (8), namely kR(S) := ıωρ0
2
Γ+(S)
˜v · n –
Γ–(S)
˜v · n
. (24) (24) Its approximated value kR(S) tends to the Rayleigh conductivity kR as 1/S as illustrated in
Fig. 5. This first-order convergence can be explained with a rigorous analysis of the solu-
tion of problem (6b)–(6d) towards infinity using the Mellin transform [16] and showing
that the solution of this problem on Γ±(S) is a superposition of a radial expansion with
respect to 1/S and of a cartesian expansion with terms decaying exponentially with re-
spect to the distance to the boundary. Similar analyses were performed for the Poisson
and Helmholtz problems in conical domains with a rough periodic boundary [6] or per-
forated wall [7]. Its approximated value kR(S) tends to the Rayleigh conductivity kR as 1/S as illustrated in
Fig. 5. This first-order convergence can be explained with a rigorous analysis of the solu-
tion of problem (6b)–(6d) towards infinity using the Mellin transform [16] and showing
that the solution of this problem on Γ±(S) is a superposition of a radial expansion with
respect to 1/S and of a cartesian expansion with terms decaying exponentially with re-
spect to the distance to the boundary. 3.1 Numerical computation of kR Similar analyses were performed for the Poisson
and Helmholtz problems in conical domains with a rough periodic boundary [6] or per-
forated wall [7]. As, more precisely, the Rayleigh conductivity kR can be expanded in powers of 1/S we
use an extrapolation in 1/S of first order approximations kR(S) for different truncation
radia S to obtain a second or higher order approximation of the limit value kR. ( 2018) 8:15 Page 12 of 20 Schmidt et al. Journal of Mathematics in Industry Figure 5 Convergence of the real and imaginary parts of the approximated Rayleigh conductivity kR(S) to its
limit value kR = 4.513 – 1.210ı in dependence of the truncation radius S for the liner DC006 at frequency
f = 306 Hz (see Table 1) Figure 5 Convergence of the real and imaginary parts of the approximated Rayleigh conductivity kR(S) to its
limit value kR = 4.513 – 1.210ı in dependence of the truncation radius S for the liner DC006 at frequency
f = 306 Hz (see Table 1) Table 1 Liner configurations. The length of the liner is L = 60 mm. The value of the viscosity is
ν(δ) = 1.4660 × 10–5 m2/s. For all these configurations b = 0.5√a
Config. Number of holes
(longitudinal,
azimuthal)
Longitudinal
inter-hole distance
δ/√a (mm)
Azimuthal
inter-hole distance
√aδ (mm)
Hole
diameter
d0δ2 (mm)
Liner
thickness
h0δ2 (mm)
σ
%
DC006
(7, 52)
8.5
8.45
1
1
1.1
DC007
(3, 20)
22
21.99
2.5
1
1.0
DC008
(7, 52)
8.5
8.45
2.5
1
6.8
DC009
(3, 20)
22
21.99
1
1
0.2 Table 1 Liner configurations. The length of the liner is L = 60 mm. The value of the viscosity is
ν(δ) = 1.4660 × 10–5 m2/s. For all these configurations b = 0.5√a For the particular case of a straight cylindrical hole that is without loss of generality
centered at y = z = 0, the domain
Ω(S) is invariant under rotation around the r axis as
well as the solution of the problem (6b)–(6d) for the near field profiles. Hence, the finite
element method in two dimensions can be used for the numerical resolution in a 2D axis-
symmetry setting. To resolve the boundary layer of size √ν0/ω on the wall boundary (cf. [5, Sect. 3.1]) we use the hp-adaptive strategy of Schwab and Suri [17] (see the mesh shown
in Fig. 3(b)). 3.1 Numerical computation of kR As Melling model uses an effective viscosity taking into account thermal
conductivity losses we show the impedance for our model with the effective viscosity as well Figure 6 Real and imaginary parts of kR in dependence of the frequency f = ω
2π Figure 6 Real and imaginary parts of kR in dependence of the frequency f = ω
2π Figure 7 Comparison of the impedance with the normalized specific resistance (left) and the normalized
specific reactance (right) computed by our model and Melling model as function of the frequency f = ω
2π for
the liner configuration DC006. As Melling model uses an effective viscosity taking into account thermal
conductivity losses we show the impedance for our model with the effective viscosity as well Figure 7 Comparison of the impedance with the normalized specific resistance (left) and the normalized
specific reactance (right) computed by our model and Melling model as function of the frequency f = ω
2π for
the liner configuration DC006. As Melling model uses an effective viscosity taking into account thermal
conductivity losses we show the impedance for our model with the effective viscosity as well In Fig. 6, we plot the effective Rayleigh conductivity kR as a function of the frequency
f := ω
2π for different liner configurations given in Table 1. As expected, following the re-
mark on the normalized specified acoustic impedance ζ, the real part of kR is positive and
its imaginary part is negative. One can also remark that for liner configurations DC006
and DC007, that have a close value of the porosity σ but quite different hole repartition
and hole diameter, their Rayleigh conductivities differ significantly in both their real and
imaginary part. In Fig. 7, we show the computed normalized specific acoustic impedance ζ for the
liner configuration DC006 in comparison with the Melling model (see [19] and [18, Eq. (12)]), that is given an analytic formula. For the latter an effective kinematic viscosity
˜ν(δ) := 2.179ν(δ) is used that shall incorporate also thermal conductivity losses near a
highly conducting wall, see [20, p. 239] and [1, p. 62]. We plotted the Rayleigh conduc-
tivities obtained from our model with this effective kinematic viscosity. The reactance
predicted by the two models are very close, where the resistence differs by up to 20%. 3.1 Numerical computation of kR The
importance of taking the thermal conductivity losses into account will be seen in compar-
ison with the measurements and be discussed later in Sect. 3.3. 3.1 Numerical computation of kR For four liner configurations, see Table 1, from experimental studies [1, 18] we have
computed the near field velocity and pressure profiles and so the effective Rayleigh con-
ductivity. The relative kinematic viscosity ν0 is computed as quotient of the kinematic vis-
cosity ν = 1.4660 × 10–5 m2/s of air at 15◦C divided by the period δ to the power of four. In Fig. 3(b) and Fig. 3(c) we illustrate the near field pressure and velocity profiles ˜p and ˜v
for the liner DC006 at frequency 306 Hz using a truncation radius S = 40. It is visible that
the pressure decays almost linearly inside the cylindrical hole, but also the behaviour at
distance to the hole. Moreover, the pressure shows close to the rim of the cylinder an edge
singularity (i.e., a corner singularity for the 2D axis-symmetric problem) that is resolved
numerically by the hp-adaptive refinement strategy. The near velocity profile shows a flux
from all sides to and through the hole. It appears that the outward flux of the imaginary
part of ˜v over Γ+(S) is negative (resp. positive over Γ–(S)) corresponding to a positive real
part of the approximate Rayleigh conductivity kR(S) (see (24)) and so of the Rayleigh con-
ductivity kR. This is in line with the inviscid case, where kR is real and positive. Moreover,
we see the higher velocity amplitude inside the hole that decays towards its boundaries. This boundary layer phenomena is more visible for lower frequencies (see Fig. 3(d)), where
one also see a local change of the velocity direction on the wall boundary. ( 2018) 8:15 Page 13 of 20 Schmidt et al. Journal of Mathematics in Industry Figure 6 Real and imaginary parts of kR in dependence of the frequency f = ω
2π
Figure 7 Comparison of the impedance with the normalized specific resistance (left) and the normalized
specific reactance (right) computed by our model and Melling model as function of the frequency f = ω
2π for
the liner configuration DC006. 3.2 Dissipation losses in acoustic ducts
3.2.1 Experimental setup and analysis The experimental study is performed in the duct acoustic test rig with a circular cross-
section (DUCT-C) at the German Aerospace Center in Berlin at ambient conditions. The
setup of the test rig is illustrated in Fig. 8. It allows high precision acoustic measurements ( 2018) 8:15 Page 14 of 20 Schmidt et al. Journal of Mathematics in Industry Figure 8 Schematic setup of the Duct Acoustic Testrig (DUCT-C) with speakers A and B, and microphones
1–12. The anechoic terminations at both ends are not shown Figure 8 Schematic setup of the Duct Acoustic Testrig (DUCT-C) with speakers A and B, and microphones
1–12. The anechoic terminations at both ends are not shown of the damping performance of various liner configurations, including grazing and bias
flow. The test duct consists of two symmetric measurement sections (section 1 and section 2
in Fig. 8) of 1200 mm length each. They have a circular cross-section with a radius Rd of
70 mm. In order to minimize the reflection of sound at the end of the duct back into the
measuring section the test duct is equipped with anechoic terminations at both ends (not
shown in Fig. 8). Their specifications follow the ISO 5136 standard. The damping module
is a chamber of 60 mm. It has a circular cross-section with a radius of 120 mm. A total of 12 microphones are mounted flush with the wall of the test duct. They are in-
stalled at different axial positions upstream and downstream of the damping module and
are distributed exponentially with a higher density towards the damping module. Two mi-
crophones are installed opposite of each other at the same axial position close to the signal
source. As evanescent modes become more prominent in the vicinity of the source, their
influence is reduced significantly by using the average value of these two microphones for
the analysis. This technique helps to reduce the errors for frequencies approaching the
cut-on frequency of the first higher order mode and thus, extends the frequency range for
accurate results. At the end of each section a loudspeaker is mounted at the circumference of the duct
(A and B in Fig. 8). They deliver the test signal for the damping measurements. The sig-
nal used here is a multi-tone sine signal. All tonal components of the signal are in the
plane wave range. 3.2 Dissipation losses in acoustic ducts
3.2.1 Experimental setup and analysis The signal has been calibrated in a way that the amplitude of each tonal
component inside the duct is about 102 dB. The microphones used in these measurements are 1/4′′ G.R.A.S. type 40BP condenser
microphones. Their signals are recorded with a 16 track OROS OR36 data acquisition
system with a sampling frequency of 8192 Hz. The source signals for the loudspeakers
are recorded on the remaining tracks. The test signal is produced by an Agilent 33220A
function generator. The signals are fed through a Dynacord L300 amplifier before they
power the Monacor KU-516 speakers. For each configuration two different sound fields are excited consecutively in two sep-
arate measurements (index a and b). Speaker A is used in the first measurement and in
the second measurement the same signal is fed into speaker B. Then, the data of section
1 and section 2 (index 1 and 2) are analyzed separately. This results in four equations for
the complex sound pressure amplitudes for each section and measurement for j = a,b: ˆp1j(z) = ˆp+
1jeıωz/c + ˆp–
1je–ıωz/c,
(25a)
ˆp2j(z) = ˆp+
2jeıω(z–L)/c + ˆp–
2je–ıω(z–L)/c,
(25b) ˆp1j(z) = ˆp+
1jeıωz/c + ˆp–
1je–ıωz/c,
(25a)
ˆp2j(z) = ˆp+
2jeıω(z–L)/c + ˆp–
2je–ıω(z–L)/c,
(25b) ˆp1j(z) = ˆp+
1jeıωz/c + ˆp–
1je–ıωz/c,
ˆp2j(z) = ˆp+
2jeıω(z–L)/c + ˆp–
2je–ıω(z–L)/c, (25a) (25b) ( 2018) 8:15 Page 15 of 20 Schmidt et al. Journal of Mathematics in Industry ˆp+ and ˆp– are the complex amplitudes of the downstream and upstream traveling
waves. The recorded microphone signals are transformed into the frequency domain using the
method presented by Chung [21]. This method rejects uncorrelated noise, e.g., turbulent
flow noise, from the coherent sound pressure signals. Therefore, the sound pressure spec-
trum of one microphone is determined by calculating the cross-spectral densities between
three signals, where one signal serves as a phase reference. In our case the phase refer-
ence signal is the source signal of the active loudspeaker. As a result we obtain a phase-
correlated complex sound pressure spectrum for each microphone signal. According to Eqs. (25a)–(25b) the measured acoustic signal is a superposition of two
plane waves traveling in opposite direction. In order to determine the downstream and
upstream propagating portions of the wave in each section, a mathematical model is fit-
ted to the acoustic microphone data. This model considers viscous and thermal conduc-
tivity losses at the duct wall. 3.2 Dissipation losses in acoustic ducts
3.2.1 Experimental setup and analysis They are included in the wave number with the following
attenuation factor α as proposed by Kirchhoff[22]: α =
1
cRd
νω
2
1 + γ – 1
√
Pr
(26) (26) with the duct radius r, the speed of sound c, the kinematic viscosity ν, the angular fre-
quency ω (as in Eqs. (3a)–(3d)), the heat capacity ratio γ , and the Prandtl number Pr. As
a result of this least-mean-square fit, the four complex sound pressure amplitudes ˆp+
1, ˆp–
1,
ˆp+
2 and ˆp–
2 are identified at position z = 0 for both measurements. These sound pressure
amplitudes are related to each other via the reflection and transmission coefficients of the
test object. This is illustrated in Fig. 9 for the two different measurements A and B. In
order to calculate the reflection and transmission coefficients r+, r–, t+, and t– from the
sound pressure amplitudes the following four relations can be derived for j = a,b: ˆp–
1j = r+ˆp+
1j + t–ˆp–
2j,
(27a)
ˆp+
2j = r–ˆp–
2j + t+ˆp+
1j. (27b) ˆp–
1j = r+ˆp+
1j + t–ˆp–
2j,
ˆp+
2j = r–ˆp–
2j + t+ˆp+
1j. (27a) (27b) Figure 9 Illustration of the sound filed in the duct for
measurements A and B by means of the sound pressure
amplitudes ˆp, the reflection coefficient r, the transmission
coefficient t, and the end reflection re Figure 9 Illustration of the sound filed in the duct for
measurements A and B by means of the sound pressure
amplitudes ˆp, the reflection coefficient r, the transmission
coefficient t, and the end reflection re Figure 9 Illustration of the sound filed in the duct for ( 2018) 8:15
Page 16 of 20 Schmidt et al. Journal of Mathematics in Industry ( 2018) 8:15
Page 16 of 20 ( 2018) 8:15 Page 16 of 20 Schmidt et al. 3.2 Dissipation losses in acoustic ducts
3.2.1 Experimental setup and analysis Journal of Mathematics in Industry The equations from both measurements are combined and solved for the reflection r+ = ˆp–
1aˆp–
2b – ˆp–
1bˆp–
2a
ˆp+
1aˆp–
2b – ˆp+
1bˆp–
2a
,
r– = ˆp+
2bˆp+
1a – ˆp+
2aˆp+
1b
ˆp+
1aˆp–
2b – ˆp+
1bˆp–
2a
(28) (28) nd transmission coefficients and transmission coefficients t+ = ˆp+
2aˆp–
2b – ˆp+
2bˆp–
2a
ˆp+
1aˆp–
2b – ˆp+
1bˆp–
2a
,
t– = ˆp+
1aˆp–
1b – ˆp+
1bˆp–
1a
ˆp+
1aˆp–
2b – ˆp+
1bˆp–
2a
(29) (29) in downstream and upstream direction, respectively. The advantage of combining the two
measurements is that the resulting coefficients are independent from the reflection of
sound at the duct terminations. These end-reflections are contained in the sound pres-
sure amplitudes, but do not need to be calculated explicitly. Moreover in the case of a
uniform and stagnant flow these coefficients do not depend on the direction we consider,
i.e., r– = r+ and t– = t+. The dissipation of acoustic energy is expressed by the dissipation coefficient. The dis-
sipation coefficient can be calculated directly from the reflection coefficient R and the
transmission coefficient T via an energy balance R± + T± + ± = 1. (30) (30) R± + T± + ± = 1. To compute these coefficients, the integration of the acoustic energy flux in a uniform
and stagnant flow yields a relation between the acoustic pressure p and acoustic power P
quantities(see Blokhintsev [23] and Morfey [24]): P± = πR2
d
2ρ0c
ˆp±2. (31 P± = πR2
d
2ρ0c
ˆp±2. (31) Then, the energy coefficients can be given relative to the pressure coefficients as: Then, the energy coefficients can be given relative to the pressure coefficients as: R+ = P–
1
P+
1
=
r+2,
R– = P+
2
P–
2
=
r–2,
T+ = P+
2
P+
1
=
t+2,
T– = P–
1
P–
2
=
t–2, R+ = P–
1
P+
1
=
r+2,
(32a)
R– = P+
2
P–
2
=
r–2,
(32b)
T+ = P+
2
P+
1
=
t+2,
(32c)
T– = P–
1
P–
2
=
t–2,
(32d) (32a) (32d) where the indices 1 and 2 refer to section 1 and section 2 of the duct as illustrated in Fig. 9. 3.3 Numerical results and comparison with experimental data Figure 10 shows the average dissipation of the different liner configurations (see Table 1)
in the DUCT-C setup (see Fig. 9) as a function of the frequency. The average dissipa-
tion represents a mean value of the dissipation results for the upstream and downstream
acoustic incidence (see Sect. 3.2.1). In a symmetric setup and without grazing flow this is,
of course, equal to the dissipation from either side of excitation. The graphs compare the
experimental values (symbols), the former theoretical model from Melling [19] (dashed
lines) and the here introduced asymptotic model (solid lines). In result, the asymptotic
model indicates a better comparison to the experimental values especially for the con-
figurations DC006 (Fig. 10(a)) and DC008 (Fig. 10(c)) where the Melling model slightly
underestimates the dissipation in the frequency range above approximately 400 Hz. For
the configuration DC007 (Fig. 10(b)) with a porosity of 1.0% and a hole diameter of 2.5 mm
both models (Melling and asymptotic) underestimate the maximum dissipation of approx-
imately 0.4 around 400 Hz revealed in the experimental studies. 3.2 Dissipation losses in acoustic ducts
3.2.1 Experimental setup and analysis The pairs (βj,ψj) and (β′
j,ψ′
j) are solution of a “2D” transverse eigen-
value problem in the wave-guide and liner parts, using the fact that the source term pinc
and the geometry are independent of the angle θ. From the mode matching and assuming
that there is only one propagative mode inside the waveguide, i.e., βj ∈ıR for j ̸= 0, the
energy dissipation coefficient is computed as := 1 –
α+
0
2 +
α–
0
2
,
(35) := 1 –
α+
0
2 +
α–
0
2
, (35) and corresponds to the energy dissipation coefficient D± (Eq. (33)) if both grazing and bias
flows are absent. 3.2 Dissipation losses in acoustic ducts
3.2.1 Experimental setup and analysis With the energy balance (30) follows the definition of the energy dissipation coefficient where the indices 1 and 2 refer to section 1 and section 2 of the duct as illustrated in Fig. 9. With the energy balance (30) follows the definition of the energy dissipation coefficient = 1 –
r±2 +
t±2
. (33) = 1 –
r±2 +
t±2
. (33) This is an integral value of the acoustic energy that is absorbed while a sound wave is
passing the damping module. The dissipation coefficient is used to evaluate the damping
performance of the test object. ( 2018) 8:15
Page 17 of 20 ( 2018) 8:15 Page 17 of 20 Schmidt et al. Journal of Mathematics in Industry 3.2.2 Numerical simulation of dissipation losses 3.2.2 Numerical simulation of dissipation losses This setup is also simulated numerically using the equivalent problem (22a)–(22b) for the
pressure with a source term corresponding to an incoming field pinc(r,θ,z) = exp(ıωz/c)
from the left. The scattered field is computed numerically using the mode matching proce-
dure with N = 5 modes [25]: we seek for the scattered field p0 under the form (see Fig. 2(b)) p0(r,θ,z) = pinc(r,θ,z) +
N–1
j=0
α–
j ψj(r) exp(–ı βj z),
z < 0,
(34a) p0(r,θ,z) = pinc(r,θ,z) +
N–1
j=0
α–
j ψj(r) exp(–ı βj z),
z < 0,
(34a)
N–1 (34a) p0(r,θ,z) =
N–1
j=0
α+
j ψj(r) exp(ı βj z),
z > L,
(34b) (34b) de the waveguide part, and under the form inside the waveguide part, and under the form p0(r,θ,z) =
2N–1
j=0
ψ′
j(r)
α′
j
+ exp
ı β′
j z
+ α′
j
– exp
ı β′
j (L – z)
,
0 < z < L,
(34c) (34c) inside the duct part. The pairs (βj,ψj) and (β′
j,ψ′
j) are solution of a “2D” transverse eigen-
value problem in the wave-guide and liner parts, using the fact that the source term pinc
and the geometry are independent of the angle θ. From the mode matching and assuming
that there is only one propagative mode inside the waveguide, i.e., βj ∈ıR for j ̸= 0, the
energy dissipation coefficient is computed as inside the duct part. 4 Conclusions We
decoupled in a systematic way the effects at different scales and derived impedance con-
ditions for the macroscopic pressure or velocity based on a proper matching of pressure
and velocity at the different scales. In difference to a direct numerical solution for acoustic
liners the overall computation effort is separated into a precomputation of the effective
Rayleigh conductivity impedance conditions, where no holes have to be resolved anymore
by a finite element mesh. The comparison with measurements in the duct acoustic test
rig with a circular cross-section at the German Aerospace Center in Berlin show that the
dissipation losses based on the impedance conditions with effective Rayleigh conductiv-
ity are well predicted. The derivation of the impedance conditions do not depend on the
cylindrical shape of the liner and can be used for others shapes like rectangular profiles. The procedure for the computation of the effective Rayleigh conductivity can not only be
extented to include thermic effects that are currently only heuristically incorporated, but
also nonlinear effects inside the hole that lead to an interaction of frequencies. Acknowledgements
The authors would like to thank Claus Lahiri (Rolls-Royce) for fruitful discussions. The research was partly conducted
during the stay of the first and second author at the TU Berlin and the first author at BTU Cottbus-Senftenberg. 4 Conclusions It has been shown that impedance conditions with one numerically computed parameter—
the effective Rayleigh conductivity—can predict well the dissipation losses of acoustic ( 2018) 8:15 Page 18 of 20 Schmidt et al. Journal of Mathematics in Industry Figure 10 Average dissipation from experiments and numerical modelling plotted over the frequency
comparing models for DC006, DC007, DC008, DC009 Figure 10 Average dissipation from experiments and numerical modelling plotted over the frequency
comparing models for DC006, DC007, DC008, DC009 liners. The effective Rayleigh conductivity can be obtained by solving numerically an in-
stationary Stokes problem in frequency domain of one hole with a scaled viscosity in an
characteristic infinite domain with prescribed pressure at infinity. For the computation
the infinite domain is truncated, where we propose approximative boundary conditions
on the artificial boundaries and an extrapolation procedure to save computation time. We
decoupled in a systematic way the effects at different scales and derived impedance con-
ditions for the macroscopic pressure or velocity based on a proper matching of pressure
and velocity at the different scales. In difference to a direct numerical solution for acoustic
liners the overall computation effort is separated into a precomputation of the effective
Rayleigh conductivity impedance conditions, where no holes have to be resolved anymore
by a finite element mesh. The comparison with measurements in the duct acoustic test
rig with a circular cross-section at the German Aerospace Center in Berlin show that the
dissipation losses based on the impedance conditions with effective Rayleigh conductiv-
ity are well predicted. The derivation of the impedance conditions do not depend on the
cylindrical shape of the liner and can be used for others shapes like rectangular profiles. The procedure for the computation of the effective Rayleigh conductivity can not only be
extented to include thermic effects that are currently only heuristically incorporated, but
also nonlinear effects inside the hole that lead to an interaction of frequencies. liners. The effective Rayleigh conductivity can be obtained by solving numerically an in-
stationary Stokes problem in frequency domain of one hole with a scaled viscosity in an
characteristic infinite domain with prescribed pressure at infinity. For the computation
the infinite domain is truncated, where we propose approximative boundary conditions
on the artificial boundaries and an extrapolation procedure to save computation time. Author details
1 1Fachbereich Mathematik, AG Numerik und Wissenschaftliches Rechnen, Technische Universität Darmstadt, Darmstadt,
Germany. 2Institut für Mathematik, Technische Universität Berlin, Berlin, Germany. 3German Aerospace Center, Institute
of Propulsion Technology, Berlin, Germany. Received: 6 January 2018 Accepted: 10 December 2018 Received: 6 January 2018 Accepted: 10 December 2018 Received: 6 January 2018 Accepted: 10 December 2018 Competing interests The authors declare that they have no competing interests. Authors’ contributions The impedance conditions are mainly derived by the first three authors that also conducted the numerical studies and
wrote the associated sections of the article. The fourth author contributed mainly with the description of the
measurements of the studied liners. All authors participated on the design of the numerical studies and the comparison
with the measurements. All authors read and approved the final manuscript. ( 2018) 8:15 ( 2018) 8:15 ( 2018) 8:15 Schmidt et al. Journal of Mathematics in Industry Funding Funding
The research was supported by Einstein Center for Mathematics Berlin via the research center MATHEON, Mathematics for
Key Technologies, in Berlin as well as the Brandenburgische Technische Universität Cottbus-Senftenberg through the
Early Career Fellowship of the second author. References On the homogenization of the Helmholtz problem with thin perforated walls of
finite length. ESAIM: Math Model Numer Anal. 2018;52(1):29–67. finite length. ESAIM: Math Model Numer Anal. 2018;52(1):29–67. 8. Delourme B, Schmidt K, Semin A. On the homogenization of thin perforated walls of finite length. Asymptot Ana
2016;97(3–4):211–64. 8. Delourme B, Schmidt K, Semin A. On the homogenization of thin perforated walls of finite length. Asymptot Anal. 2016;97(3–4):211–64. 9. Semin A, Schmidt K. On the homogenization of the acoustic wave propagation in perforated ducts of finite leng
for an inviscid and a viscous model. Proc R Soc Lond A. 2018;474(2210):20170708. 9. Semin A, Schmidt K. On the homogenization of the acoustic wave propagation in perforated ducts of finite length
for an inviscid and a viscous model. Proc R Soc Lond A. 2018;474(2210):20170708. 10. Schmidt K, Thöns-Zueva A, Joly P. Asymptotic analysis for acoustics in viscous gases close to rigid walls. Math Mo
Methods Appl Sci. 2014;24(9):1823–55. 10. Schmidt K, Thöns-Zueva A, Joly P. Asymptotic analysis for acoustics in viscous gases close to rigid walls. Math Models
Methods Appl Sci. 2014;24(9):1823–55. 10. Schmidt K, Thöns-Zueva A, Joly P. Asym
Methods Appl Sci. 2014;24(9):1823–55. 11. Goldstein CI. A finite element method for solving Helmholtz type equations in waveguides and other unbounded
domains. Math Comput. 1982;39(160):309–24. 11. Goldstein CI. A finite element method for solvin
domains. Math Comput. 1982;39(160):309–24. 12. Sanchez-Hubert J, Sánchez-Palencia E. Acoustic fluid flow through holes and permeability of perforated walls. J Mat
Anal Appl. 1982;87(2):427–53. 12. Sanchez-Hubert J, Sánchez-Palencia E. Acoustic fluid flow through holes and permeability of perforated walls. J Mat
Anal Appl. 1982;87(2):427–53. pp
13. Webster AG. Acoustical impedance and the theory of horns and of the phonograph. Proc Natl Acad Sci USA. 1919;5(7):275–82. pp
13. Webster AG. Acoustical impedance and the theory of horns and of the phonograph. Proc Natl Acad Sci USA. 1919;5(7):275–82. 14. Bendali A, Fares M, Laurens S, Tordeux S. Numerical study of acoustic multiperforated plates. ESAIM Proc. 2012;37:166–77. 15. Bendali A, Fares M, Piot E, Tordeux S. Mathematical justification of the Rayleigh conductivity model for perforat
plates in acoustics. SIAM J Numer Anal. 2013;73(1):438–59. p
16. Kozlov VA, Maz’ya VG, Rossmann J. Elliptic boundary value problems in domains wi
Mathematical surveys and monographs. vol. 52. Providence: Am. Math. Soc.; 1997. p
16. Kozlov VA, Maz’ya VG, Rossmann J. Elliptic boundary value problems in domains with point singularities. Abbreviation Abbreviation
Not applicable. References References
1. Lahiri C. Acoustic performance of bias flow liners in gas turbine combustors [PhD thesis]. Berlin, Germany: Technische
l
h
d
b l
d
h
dl References
1. Lahiri C. Acoustic performance of bias flow liners in gas turbine combustors [PhD thesis]. Berlin, Germany: Technische References
1. Lahiri C. Acoustic performance of bias flow liners in gas turbine combustors [PhD thesis]. Berlin, Germany: Technische
Uni ersität Berlin 2014 https //depositonce t
berlin de/handle/11303/4567 1. Lahiri C. Acoustic performance of bias flow liners in gas turbine combustors [PhD thesis]. Berlin, Germany: Technische
Universität Berlin; 2014. https://depositonce.tu-berlin.de/handle/11303/4567. 1. Lahiri C. Acoustic performance of bias flow liners in gas turbine combustors [PhD thesis]. Berlin, Germany: Technische
Universität Berlin; 2014. https://depositonce.tu-berlin.de/handle/11303/4567. Universität Berlin; 2014. https://depositonce.tu-berlin.de/handle/11303/4567. Universität Berlin; 2014. https://depositonce.tu-berlin.de/handle/11303/4567. p
p
2. Rayleigh JWS. On the theory of resonance. Philos Trans R Soc Lond. 1871;161:77–118. 2. Rayleigh JWS. On the theory of resonance. Philos Trans R Soc Lon 3. Rayleigh JWS. The theory of sound. vol. 2. New York: Dover; 1945 3. Rayleigh JWS. The theory of sound. vol. 2. New York: Dover; 1945. 4. Semin A, Schmidt K. Absorbing boundary conditions for the viscous acoustic wave equation. Math Methods App
Sci. 2016;39(17):5043–65. 4. Semin A, Schmidt K. Absorbing boundary conditions for the viscous acoustic wave equation. Math Methods Appl
Sci. 2016;39(17):5043–65. 5. Popie V. Modélisation asymptotique de la réponse acoustique de plaques perforées dans un cadre linéaire avec 5. Popie V. Modélisation asymptotique de la réponse acoustique de plaques perforées dans un cadre linéaire avec
étude des effets visqueux [PhD thesis] Toulouse France: Université de Toulouse; 2016 5. Popie V. Modélisation asymptotique de la réponse acoustique de plaques perforées dans
étude des effets visqueux [PhD thesis]. Toulouse, France: Université de Toulouse; 2016. 5. Popie V. Modélisation asymptotique de la réponse acoustique de plaques perforées dan
étude des effets visqueux [PhD thesis]. Toulouse, France: Université de Toulouse; 2016. p
y
p
q
p
q
p q
p
étude des effets visqueux [PhD thesis]. Toulouse, France: Université de Toulouse; 2016. étude des effets visqueux [PhD thesis]. Toulouse, France: Université de Toulouse; 2016. 6. Nazarov SA. The Neumann problem in angular domains with periodic and parabolic perturbations of the boundary. Tr Mosk Mat Obˆs. 2008;69:182–241. 6. Nazarov SA. The Neumann problem in angular domains with periodic and parabolic perturbations of the boundary
Tr Mosk Mat Obˆs. 2008;69:182–241. 7. Semin A, Delourme B, Schmidt K. Acknowledgements
h
h
ld l k Page 19 of 20 Publisher’s Note
S
i
N
i Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Schmidt et al. Journal of Mathematics in Industry ( 2018) 8:15 Schmidt et al. Journal of Mathematics in Industry 24. Morfey CL. Acoustic energy in non-uniform flows. J Sound Vib. 1971;14(2):159–70. 23. Blokhintsev DI. Acoustics of a nonhomogeneous moving medium. NACA Technical Memorandum 1399, 25. Semin A, Thöns-Zueva A, Schmidt K. Simulation of reflection and transmission properties of multiperforated acoustic
liners. In: Quintela P, et al, editors. Progress in industrial mathematics at ECMI 2016. Mathematics in industry. vol. 26.
Cham: Springer; 2017. p. 69–76. References Mathematical surveys and monographs vol 52 Providence: Am Math Soc ; 1997 Mathematical surveys and monographs. vol. 52. Providence: Am. Math. Soc.; 199 17. Schwab C, Suri M. The p and hp versions of the finite element method for problems with boundary layers. Math
Comput. 1996;65(216):1403–30. 17. Schwab C, Suri M. The p and hp versions of the finite element method for problems with boundary layers. Math
Comput. 1996;65(216):1403–30. 18. Lahiri C, Bake F. A review of bias flow liners for acoustic damping in gas turbine combustors. J Sound Vib. 2017;400:564–605. 18. Lahiri C, Bake F. A review of bias flow liners for acoustic damping in gas turbine combustors. J Sound Vib. 2017;400:564–605. 19. Melling TH. The acoustic impendance of perforates at medium and high sound pressure levels. J Sound Vib. 1973;29(1):1–65. 19. Melling TH. The acoustic impendance of perforates at medium and high sound pressure levels. J Sound Vib. 1973;29(1):1–65. 20. Crandall IB. Theory of vibrating systems and sound. New York: Van Nostrand; 1926. 20. Crandall IB. Theory of vibrating systems and sound. New York: Van Nostrand; 1926. 21. Chung JY. Rejection of flow noise using a coherence functio 22. KirchhoffG. Über den Einfluss der Wärmeleitung in einem Gase auf die Schallbewegung. Ann Phys Chem. 1868;210(6):177–93. Page 20 of 20 Page 20 of 20 23. Blokhintsev DI. Acoustics of a nonhomogeneous moving medium. NACA Technical Memorandum 1399,
Washington DC; 1956 Originally published 1946 in Russian language ( 2018) 8:15 ( 2018) 8:15
|
https://openalex.org/W1945070435
|
https://revistas.udesc.br/index.php/palindromo/article/download/4228/2988
|
Portuguese
| null |
Possíveis Leituras de Extensões da Arte nas Ilustrações de Livros: a Pós-Produção nas Imagens de Rébecca Dautremer
|
Palíndromo
| 2,014
|
cc-by
| 7,074
|
PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC 1 Professora do Departamento de Design da Universidade do Estado de Santa Catarina. Resumo Nesse artigo é apresentada uma proposta de leitura de imagens a partir do livro ilustrado
associada à leitura de manifestações artísticas, com foco na formação visual na escola. É
tomado como instrumento de leitura o modelo desenvolvido por Ramalho e Oliveira (2005). Este modelo parte da Semiótica Greimasiana e baseia-se na desconstrução e reelaboração
das imagens e na exploração de suas significações. Como forma de experimentação desta
proposta é apresentado um breve exercício de leitura de ilustrações do livro Diario Segreto
di Pollicino (2010), de Rébecca Dautremer, relacionadas à perspectiva da Arte na Pós-
Produção segundo características levantadas por Bourriaud (2009). Palavras-Chave: Livro Ilustrado. Leitura de imagens. Pós-Produção Possíveis leituras de extensões da arte nas ilustrações de livros:
a Pós-Produção nas imagens de Rébecca Dautremer Anelise Zimmermann1
Universidade do Estado de Santa Catarina Keywords: Picturebook. Picture reading. Post-production. Introdução Vivendo em meio ao que Calvino (1990) preconizou como o Milênio da Visibilidade,
envolvidos pelo "dilúvio das imagens pré-fabricadas", somos alertados: "Em nossa memória
se depositam, por estratos sucessivos, mil estilhaços de imagens, semelhantes a um
depósito de lixo, onde é cada vez menos provável que uma delas adquira relevo". (Ibid., p. 107). A partir de outra perspectiva Bourriaud (2009) chama a atenção para o fato de que
"não estamos saturados de imagens; estamos submetidos à escassez de certas imagens"
(2009, p. 59). Segundo Bourriaud (Ibid.) cabe à arte contemporânea a elaboração de
imagens que despertem o observador para novos enredos e leituras do mundo que o cerca;
imagens estas que se oponham, que proponham, que desconstruam e revelem múltiplas
combinações, envolvendo, provocando e convocando seu leitor a sua reelaboração. Ramalho e Oliveira (2005), entretanto, destaca o fato de que essa participação do
observador, de fato ativa, só é possível quando este possui os recursos mínimos para uma
leitura crítica das imagens, podendo assim ultrapassar os níveis superficiais das
significações. É neste contexto que encontram-se os livros ilustrados, livros compostos por textos
verbais e visuais que, em conjunto, propõem uma história. Estes livros, geralmente
associados ao público infantil, muitas vezes são abandonados tão logo a alfabetização
verbal é considerada completa, ignorando possíveis participações das ilustrações na
elaboração das histórias, como que sendo desnecessárias. Entretanto, a ilustradora
Dautremer ao falar sobre seu trabalho comenta: "Com minhas ilustrações tento construir
uma história que não é redundante ao texto. [...] Tento oferecer uma outra perspectiva sobre
a aventura, levar o leitor a um caminho paralelo que irá enriquecer a sua leitura e abrir
outras portas. (DAUTREMER, 2010, p. 01, tradução nossa). Ignorando tais possibilidades
das ilustrações pais e professores muitas vezes esquecem da importância da aprendizagem
e exercício da leitura visual e sua participação na elaboração das mensagens. Com base em
tais aspectos neste artigo são discutidos os conceitos de "livro infantil" e "livro ilustrado",
bem como questões referentes ao seu público e formas frequentes de leitura. Em seguida, considerando a importância da leitura visual e sua instrumentalização, é
sugerida uma proposta de atividade de leitura de imagens por meio de ilustrações de livros e
sua aproximação com o ensino da arte. PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC Abstract Herein we present a proposal to read images from illustrated books associated with the reading of
artistic manifestations, focusing on visual training at school. It is taken as a tool for reading the model
developed by Ramalho and Oliveira (2005), which is based on Greimas Semiotics by deconstructing
and re-elaborating images to and explore its meanings. As a way of testing this proposal, we propose
a short exercise to read illustrations from Rebecca Dautremer’s Diario Segreto di Pollicino (2010),
related to the perspective of Art in Post-Production, according to characteristics raised by Bourriaud
(2009). Keywords: Picturebook. Picture reading. Post-production. PALÍNDROMO
Nº
10
/
2013
123 PALÍNDROMO
Nº
10
/
2013
123 Introdução Partindo de estudos anteriores (ZIMMERMANN;
RAMALHO e OLIVEIRA, 2007), sugere-se a utilização do modelo de leitura de imagens
proposto pela pesquisa de Ramalho e Oliveira (2005), o qual tem como base a Semiótica PALÍNDROMO
Nº
10
/
2013 PALÍNDROMO
Nº
10
/
2013 124 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC Greimasiana. Este modelo pode ser aplicado aos mais variados tipos de imagens, de forma
bastante didática e clara, portanto também na escola com as mais variadas faixas etárias,
tanto na leitura do livro ilustrado quanto na leitura de produções artísticas. Como exemplo de leitura de imagens são analisados aspectos estéticos e narrativos
das ilustrações de Rébecca Dautremer do livro Diario Segreto di Pollicino (LECHERMEIER,
2010), uma adaptação do clássico Pequeno Polegar. A análise é feita a partir da perspectiva
da Arte na Pós-Produção segundo características levantadas por Bourriaud (2009). Como
fontes de referência à análise foram incluídas à consulta ao Rébecca Dautremer: Artbook
(DAUTREMER, 2009), portfólio de sua trajetória profissional e à entrevistas concedidas pela
ilustradora a publicações do meio literário. estrutura narrativa, seja ela formada por imagens ou palavras. (RAMALHO E OLIVEIRA, 2005).
3 Estes livros foram originalmente publicados em inglês. Em Livro Ilustrado: Palavras e imagens (NIKOLAJEVA;
SCOTT, 2011) consta em nota de rodapé comentários acerca da tradução do termo empregado "livro ilustrado".
"Como no Brasil essa nomenclatura ainda é controversa, optou-se pelas seguintes traduções: picturebook para
"livro ilustrado", illustrated book, picture book e books with pictures para "livro com ilustração". Nessas três
últimas denominações, a obra exemplificada não apresenta inter-relação explícita entre palavra e imagem.
Importante notar que as autoras deste livro distinguem picturebook de picture book, diferentemente de outros
teóricos citados ao longo da obra." [Nota dos editores]. (Ibid., p. 13) 2 Considera-se que o termo texto, dentro da perspectiva da semiótica greimasiana refere-se a toda e
estrutura narrativa, seja ela formada por imagens ou palavras. (RAMALHO E OLIVEIRA, 2005).
3 2 Considera-se que o termo texto, dentro da perspectiva da semiótica greimasiana refere-se a toda e qualquer
estrutura narrativa, seja ela formada por imagens ou palavras. (RAMALHO E OLIVEIRA, 2005).
3 Estes livros foram originalmente publicados em inglês. Em Livro Ilustrado: Palavras e imagens (NIKOLAJEVA;
SCOTT, 2011) consta em nota de rodapé comentários acerca da tradução do termo empregado "livro ilustrado".
"Como no Brasil essa nomenclatura ainda é controversa, optou-se pelas seguintes traduções: picturebook para
"livro ilustrado", illustrated book, picture book e books with pictures para "livro com ilustração". Nessas três
últimas denominações, a obra exemplificada não apresenta inter-relação explícita entre palavra e imagem.
I
t
t
t
t
d
t
li
di ti
i t
b
k d
i t
b
k dif
t
t
d
t 1. O livro ilustrado Em estudos anteriores (ZIMMERMANN; OLIVEIRA, 2007) foi proposto a utilização
de livros ilustrados como recurso para o exercício da leitura visual na escola e sua
aproximação com o ensino da arte. Alguns aspectos já levantados merecem ser retomados
visto que justificam a continuação da pesquisa sobre o tema. Convém salientar a utilização
do termo "livro ilustrado" em substituição ao termo "livro infantil" empregado anteriormente
(Ibid.). A este último é frequentemente atribuído o estereótipo de livro exclusivamente para
crianças, repleto de imagens extremamente coloridas e decorativas com a função de auxiliar
a leitura do texto verbal2 àqueles que estão em período de alfabetização. Partindo desse
ponto de vista muitos jovens leitores abandonam, e muitos são encorajados a isso, os livros
ilustrados a medida que desenvolvem sua leitura verbal, ignorando porém, as possibilidades
da leitura visual através dos livros. A utilização do termo "livro ilustrado", utilizado em traduções de publicações
recentes3 (LINDEN, 2011a; SCOTT e NIKOLAJEVA, 2011) mas ainda pouco empregado no PALÍNDROMO
Nº
10
/
2013 PALÍNDROMO
Nº
10
/
2013 125 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC Brasil, parece mais adequada a esse estudo pois considera que livros com ilustrações não
se destinam exclusivamente ao público infantil, podendo falar a leitores de idades variadas. Um aspecto interessante quanto a este assunto é levantado por Linden (2011b, p. 01), segundo a qual a leitura do livro ilustrado é muitas vezes uma leitura "partilhada", ou
seja, feita conjuntamente por pais e filhos, também chamada de "dupla audiência"
(NIKOLAJEVA; SCOTT, 2011). Tal fato exige que o livro desperte o interesse destes
diferentes leitores, explorando o diálogo entre o texto verbal e o texto visual a partir de
diferentes camadas de significações. O leitor transita entre o verbal e o visual, em uma concatenação sempre
expansiva do entendimento. Cada nova leitura, tanto de palavras como de
imagens, cria pré-requisitos melhores para uma interpretação mais
adequada do todo. Presume-se que as crianças sabem disso por intuição
quando pedem que o mesmo livro seja lido para elas em voz alta repetidas
vezes. Na verdade, elas não leem o mesmo livro; elas penetram cada vez
mais fundo em seu significado. É muito comum os adultos perderem a
capacidade de ler os livros ilustrados dessa maneira, porque ignoram o todo
e encaram as ilustrações como meramente decorativas. (NIKOLAJEVA;
SCOTT, 2011, p. 1. O livro ilustrado 14) Muitas vezes, em uma leitura partilhada, a criança percebe detalhes na história
completamente ignorados pelo adulto, pois enquanto este concentra-se na leitura do texto
verbal, a criança percebe o conjunto do texto verbal - texto visual (LINDEN, 2011b). Também de um modo geral a criança se permite observar com mais atenção às ilustrações,
procurando pistas que possam estar escondidas, ora girando o livro de ponta-cabeça, ora
alterando a sequência das páginas, subvertendo a linearidade da história. Hans Christian Andersen (1805-1875), escritor considerado um dos precursores da
literatura infantil, já trabalhava com a ideia da leitura "partilhada", identificando as
possibilidades e dificuldades de compreensão desse gênero literário pelo público adulto. Segundo anotações feitas em um diário pessoal, Andersen teria ficado irritado com uma
estátua criada em sua homenagem pelo fato de ser representado rodeado por crianças: "Eu
disse [...] que os meus contos eram tanto para os adultos quanto para as crianças, e que
estas últimas compreendiam somente as personagens secundárias, e que só as pessoas
maduras viam e compreendiam tudo." (ANDERSEN, 2004, p. 18). Andersen também
comenta sobre esse mesmo assunto em uma carta escrita a seu amigo Ingemann: '"Pego
uma ideia para adultos e a conto às crianças, sempre me lembrando de que o pai e a mãe
geralmente a escutam e que é preciso dar-lhes assunto sobre o qual pensar". (Ibid., p. 18) PALÍNDROMO
Nº
10
/
2013 PALÍNDROMO
Nº
10
/
2013 126 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC Essa barreira, entre o infantil e o adulto, tem gerado muitos questionamentos sobre
os limites os definem. Um exemplo são as diferentes reações provocadas pelo livro Where
the wild things are (1963), escrito e ilustrado por Maurice Sendak (1928 - 2012). O livro,
logo após sua primeira publicação, recebeu diversos prêmios literários, entre estes o
Caldecott Medal (1964), justamente por quebrar paradigmas do gênero da literatura infantil. Entretanto, nos primeiros anos após seu lançamento, este mesmo livro foi fortemente
proibido nas bibliotecas americanas, bem como sua venda em países da América Latina
(BALDASSARRO, 2011). Às objeções à obra iam desde o comportamento de seus
personagens, considerados em alguns momentos "inadequados", podendo afetar de forma
prejudicial o comportamento de seus leitores4, até às características estéticas das
ilustrações, acusadas de "escuras e sombrias", podendo ser relacionadas à bruxaria e
elementos sobrenaturais. 4 "Um menino jogando uma garrafa foi considerado um comportamento perigoso e Sendak foi acusado de
glorificar a raiva de Max [personagem principal], o que levou os psicólogos a condená-lo como "muito escuro e
assustador." Em março de 1969, na coluna para mulheres do 'Home Journal, o psicólogo infantil Bruno
Bettelheim chamou o livro psicologicamente prejudicial para crianças de 3 a 4 anos. Segundo ele, a idéia de uma
mãe privar uma criança de alimentos era uma forma inadequada de punição, podendo traumatizar os jovens
leitores." (BALDASSARRO, 2011, p. 01) (
,
, p
)
5 Convém salientar que o conceito de infância está em constante transformação (ARIÈS, 1981), interferindo, com
isso, no que pode ser considerado infantil ou não, e mesmo adequado à criança. 1. O livro ilustrado Também a ilustradora contemporânea Dautremer (1971- ) tem suas ilustrações
muitas vezes classificadas como "sombrias", "melancólicas" e "sinistras" (DAUTREMER,
2013). Falando sobre o assunto, Dautremer comenta: “Não vejo por que não falar de
melancolia para as crianças. Eu não me coloco impedimento algum de falar disso, é meu
gosto pessoal." (Ibid., p. 01). Quanto à relação de suas ilustrações com seu público
Dautremer diz: "Não penso especificamente nas crianças quando trabalho, penso em
pessoas de todas as idades. De fato, muitos dos meus livros são comprados por adultos."
(DAUTREMER, 2011, p. 01). Reafirmando essa ideia, Dautremer ainda comenta: "Minhas
imagens são ilustrações claras, legíveis e de fácil compreensão, a partir daí cada um faz o
que quer com estas imagens. Eu não digo 'Ei, eu trabalho para as crianças.' Não mesmo. Eu
não ponho barreira entre pais e filhos. Crio imagens para pessoas ..." Entretanto, no meio literário de uma forma geral, as barreiras ou o estranhamento
entre o adulto e o infantil existem e não parecem bem resolvidas5. Em outras áreas, como
por exemplo no mercado cinematográfico, há tempos foram identificadas as possibilidades
da leitura partilhada entre crianças e adultos, investindo-se cada vez mais nas produções de
animações, lotando salas de cinema com públicos das mais variadas idades. PALÍNDROMO
Nº
10
/
2013 PALÍNDROMO
Nº
10
/
2013 127 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC 6 No caso dos livros ilustrados, mesmo existindo o "original", ou seja, a ilustração original, esta é concebida para
ser reproduzida considerando os aspectos técnicos dessa reprodução. 2. O livro ilustrado e suas possíveis leituras na escola Recuperando
argumentos
anteriores
(ZIMMERMANN;
OLIVEIRA,
2007),
é
interessante mencionar fatores relevantes à escolha do artefato “livro ilustrado” para compor
exercícios de leitura visual e suas possíveis relações com o ensino da arte na escola. Convém lembrar que o livro, por suas características físicas, possibilita a leitura individual ou
coletiva, respeitando o ritmo de seus leitores, permitindo que detalhes sejam retomados com
facilidade, por meio do manuseio simples. Além disso, o livro já está presente na escola,
sendo assim um recurso de fácil acesso ao professor, dispensando a necessidade de
reproduções ou adequações. Embora sendo um múltiplo, assim como a gravura, o livro é
uma reprodução elaborada para ser assim visualizada por seu público, não passando por
nenhum processo de distorção além de seu próprio processo de produção6. No que se refere a possíveis relações entre as ilustrações de livros ilustrados e
produções artísticas é inegável a presença das artes no repertório visual de muitos
ilustradores, a qual transparece em suas imagens, tanto por seus aspectos técnicos, como
narrativos, resgatando ora referências do passado, ora indicando características
contemporâneas (Ibid.). A ilustradora Dautremer, por exemplo, busca referências imagéticas
de pintores do século XVI e XVII, entre eles Brueghel (cerca de 1525-69), Velázquez (1599 -
1660), e Vermeer (1632 - 1675) (SOBRINO, 2011, p. 01), assim como de fotógrafos do
século XIX, como Julia Margaret Cameron (1815 -1879) ou fotógrafos contemporâneos,
entre eles Graciela Iturbide (1942 - ) e Raymond Depardon (1942- ) (CADILHAC, 2010). A proposta aqui apresentada, concentra-se nas características estéticas e
possibilidades narrativas das ilustrações, não aspirando atribuir a estas o caráter de arte ou
mesmo aprofundar a essa discussão. Também não se ignora os riscos da chamada
didatização da literatura, quando "o livro vira tarefa escolar e não um objeto de prazer"
(LINS, 2003, p. 43). O que se propõe, entretanto, é a exploração das possibilidades de
leitura dos livros, estabelecendo-se, quando adequado, relações com produções artísticas e
repertório visual de seus leitores. Para tanto sugere-se o uso da semiótica discursiva e sua vertente derivada dos
estudos de Algirdas Julien Greimas, denominada greimasiana, especificamente o modelo PALÍNDROMO
Nº
10
/
2013 PALÍNDROMO
Nº
10
/
2013 128 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC desenvolvido por Ramalho e Oliveira (2005). 7 Imagem: "consideradas toda e qualquer imagem estética produzido pelo homem, seja ela uma obra de arte ou
não; incluem-se também, além das manifestações do código visual, aquelas pertencentes ao sistema cênico,
musical, audiovisual e mesmo ao verbal. [...] Este conceito abrangente de imagem abarca ainda, sem
discriminação, tanto as produções genericamente classificadas como eruditas, quando as de origem popular e,
igualmente, as que se destinam à massa populacional, desde produtos industriais até os televisivos. (RAMALHO
e OLIVEIRA, 1998, p. 3) 2. O livro ilustrado e suas possíveis leituras na escola Tal sugestão deve-se ao fato deste modelo de
leitura ser adaptável às mais variadas mídias, imagens7 e áreas do conhecimento, podendo
ser aplicado a estudos com alunos de faixas etárias variadas, de forma clara e objetiva. O modelo de leitura de imagens proposto por Ramalho e Oliveira (2005) baseia-se
na decomposição da imagem e sua recomposição, indo das significações mais amplas às
mais específicas e complexas, em um trânsito entre os elementos. Por meio de
procedimentos relacionais buscam-se efeitos de sentido em diferentes camadas de
significações que podem muitas vezes passar despercebidos em uma leitura superficial das
imagens. São inúmeras as trilhas que se entrecruzam no visível da imagem (plano de
expressão) ao mesmo tempo em que tecem a significação (plano de
conteúdo); daí a necessidade de observar minuciosamente toda a imagem,
resgatando os pontos mais relevantes para, a partir deles, recriar,
traduzindo uma teia de elementos e procedimentos significantes que, como
tal, é construída por meio de linhas paralelas, concêntricas, todas
relacionadas. (Ibid., p. 53) A partir dessa prática de leitura é possível que um mesmo livro ilustrado conte
diferentes versões de uma história, também permitindo a leitura "partilhada”, como já
proposto por Andersen (2004). Assim, considera-se que são as experiências do autor,
materializadas por meio de sua obra, somadas às experiências do leitor que irão compor a
história. Quanto a esses aspectos Ramalho e Oliveira explica: Em cada texto visual está registrado um discurso, evidenciando uma visão
específica de seu criador, ou seja, o modo como o autor da obra vive e vê o
mundo também é mostrado na sua criação. A imagem mostra a sua visão
de mundo, suas relações com o seu contexto, além de sua capacidade de
manipulação do código ao qual pertence a imagem. (RAMALHO e
OLIVEIRA, 2005, p. 52) Vale salientar, contudo, que "tão logo o criador termine o seu trabalho, ele não mais
lhe pertence. A imagem passa a falar por si mesma, independentemente do que seu autor
tenha desejado dizer."(Ibid., loc. cit.). Tal argumento remete à fala de Dautremer citada
anteriormente, na qual reconhece a participação do leitor ao dizer "Minhas imagens são PALÍNDROMO
Nº
10
/
2013 129 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC ilustrações claras, legíveis e de fácil compreensão, a partir daí cada um faz o que quer com
estas imagens. (DAUTREMER, 2010, p. 01, tradução nossa). 3. A Pós-Produção nas ilustrações de Dautremer Em um exercício de leitura de imagens a partir da desconstrução e reconstrução de
ilustrações de Dautremer no livro Diario Segreto di Pollicino (2010), algumas articulações
entre sua obra e características da arte contemporânea se revelam. Considerando a
amplitude desta leitura, a análise apresentada a seguir concentra-se em apenas alguns
aspectos relacionados ao que o filósofo, crítico, editor e curador francês Nicolas Borriaud
define como a Arte na Pós-Produção. Também devido à extensão da obra analisada,
aproximadamente 200 páginas praticamente todas ilustradas, apenas algumas destas foram
selecionadas para compor a análise. De forma complementar são também feitas referências à publicação "Rébecca
Dautremer: Artbook" (2009), que contém parte do portfólio da ilustradora, detalhes de seu
processo criativo e supostas cenas de seu cotidiano. Este livro, escrito por Philippe
Lechermeier e ilustrado por Rébecca Dautremer, foi originalmente publicado em francês
com o título Journal secret du Petit Poucet, traduzido em português como Diário Secreto do
Pequeno Polegar. A obra apresenta um reconto do clássico O Pequeno Polegar, uma
adaptação de histórias populares do século XVII, escrito pelo francês Charles Perrault (1628
- 1703) e publicado pela primeira vez em 1697. Apesar de a primeira publicação ser também
ilustrada, foi na reedição de 1862, chamada “Contes de Perrault” que as ilustrações, feitas
por Gustave Doré, ganharam notoriedade. Nessa segunda versão o livro apresenta as
dimensões aproximadas de 19,5 x 24,5 cm, sendo impresso em preto e branco, como as
ilustrações reproduzidas através da xilogravura. O conto ocupa aproximadamente trinta e
quatro páginas do livro, apresentando 10 ilustrações, sendo que, devido às características
dos processos de impressão da época, as ilustrações encontram-se todas em páginas
isoladas, não sendo acompanhadas por texto verbal8. Já na versão de Lechermeier e Dautremer o livro apresenta um formato menor, 21 x
18 cm, entretanto é composto por aproximadamente duzentas páginas, o que faz com que
diferencie-se do formato usual de livros ilustrados, podendo ser associado ao formato de um
pequeno álbum pessoal ou diário, como o próprio título do livro indica: "Diário Secreto do PALÍNDROMO
Nº
10
/
2013 PALÍNDROMO
Nº
10
/
2013 130 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC Pequeno Polegar". As ilustrações aparecem ora intercaladas ao texto, ora misturadas a ele,
em alguns momentos o sobrepondo, interferindo em uma suposta sequência narrativa, e em
outros momentos é o texto, em forma de anotações pessoais, que se sobrepõe à ilustração. 8 Inclusive, nas páginas compostas por ilustrações, a impressão foi feita em apenas um dos lados da folh 3. A Pós-Produção nas ilustrações de Dautremer As técnicas utilizadas na construção das imagens são as mais variadas, passando por
algumas consideradas clássicas como a pintura em guache e nanquim, até técnicas mistas,
envolvendo a colagem e a fotografia. Adentrando a características mais específicas de algumas destas ilustração convém
previamente rever o que o Bourriaud (2009) define como a arte na Pós-Produção. Segundo
ele vivemos hoje um período de "caos cultural", caracterizado pela produção imensurável de
imagens e produtos dos mais variados, aos quais somos expostos constantemente. O
acesso às produções do passado nunca foi tão fácil, muitas vezes misturando-se às
produções atuais. Em meio a esse cenário surge a figura do DJ, que se aproveita das
produções já existentes para elaborar outras composições, questionando concepções de
autoria e novidade. Baseado nestes e em outros argumentos, Bourriaud propõe uma tipologia para a arte
contemporânea, caracterizando-a como a arte da Pós-Produção, da mixagem, do aplique,
do enxerto, do sampleamento e do scratching, configurando possibilidades de
reprogramação do mundo cotidiano e da arte do passado. Tal concepção tem direta relação com as possibilidades que as mídias digitais
atualmente oferecem, através das quais é possível interferir, alterar, mixar e reprogramar
mensagens já veiculadas, concebendo novos enredos. O próprio termo "Pós-Produção"
atribuído à arte por Bourriaud é sinal de apropriação, visto ter sido tomado do meio
cinematográfico, televisivo e fonográfico. Este denomina o tratamento e a reelaboração de
registros por meio de cortes, edições ou efeitos especiais, de forma a construir e reelaborar
mensagens. A ele relacionam-se termos como "reprogramar" e "recodificar", também
presentes no discurso de Bourriaud. Dentro
dessa
perspectiva
são
caracterizadas
muitos
obras
produzidas
principalmente a partir dos anos de 1990, as quais expõem o exagero e a variedade de
oferta cultural presentes no cotidiano. As bases da arte da Pós-Produção tem sua origem com as obras de ready-made de
Duchamp: a apropriação de produtos industriais pela arte e seu deslocamento, inserindo-o PALÍNDROMO
Nº
10
/
2013 131 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC em um novo contexto. "Não se trata mais de fabricar um objeto, mas de escolher entre os
objetos existentes e utilizar ou modificar o item escolhido segundo uma intenção específica"
(Ibid., p.22). A matéria-prima do artista deixa de ser a matéria bruta e passa a ser as
mercadorias, incorporando o mercado de consumo à arte. 3. A Pós-Produção nas ilustrações de Dautremer Entre exemplos na Pós-Produção
estão as obras de Michel Majerus, nas quais são utilizadas embalagens e imagens
publicitárias na construção de grandes painéis. Retomando as ilustrações de Dautremer, também em muitas delas são utilizadas
imagens de embalagens, valendo-se de humor e ironia, como no exemplo da Figura 1. As
embalagens são fictícias, porém, apresentam a estética de embalagens reais antigas. A
ironia se dá pelo fato de que, a esse ponto da história, Pollicino, o personagem que escreve
o diário, fala sobre a escassez de comida que sua família está enfrentando, o que os obriga
a comer sopa de pedra todos os dias. A representação desse cenário por meio de rótulos de
embalagem é uma escolha da ilustradora, ou seja, não consta no texto. Visualizada dessa forma, mesmo as sopas de pedra parecem poder se transformar
em produtos comerciais, ganhando embalagens nas quais seus "benefícios" são
destacados, ironizando, de certa forma, o mercado de consumo onde produtos os mais
absurdos, enchem prateleiras e sempre encontram compradores. As embalagens também aparecem em uma das fotografias de Dautremer com sua
família em uma suposta cena de seu cotidiano: o café da manhã, imagem que está presente
em seu Artbook (Figura 2). Na fotografia, produtos, embalagens e marcas misturam-se em
um cenário de caos e identificação com o consumo contemporâneo. Figura 1: Páginas 19 e 20 do Diario Segreto di Pollicino (LECHERMEIER, 2010)
Figura 2: Página 82 do Rébecca Dautremer: Artbook (DAUTREMER, 2009) Figura 1: Páginas 19 e 20 do Diario Segreto di Pollicino (LECHERMEIER, 2010)
Figura 2: Página 82 do Rébecca Dautremer: Artbook (DAUTREMER, 2009) PALÍNDROMO
Nº
10
/
2013
13 PALÍNDROMO
Nº
10
/
2013 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC Dautremer também se apropria da linguagem gráfica utilizada pela indústria para
mencionar aspectos da vida do personagem Pollicino. Ao apresentar um manual de
montagem de um mobiliário que comporta seis camas empilhadas, Dautremer expõe o fato
de que Pollicino mora em uma casa tão pequena e com tantos moradores que seria inviável
comportar uma cama para cada um (Figura 3). Em seu Artbook (2009) Dautremer revela
referências utilizadas na elaboração dessa ilustração, as quais, de fato, advém de manuais
de produtos industriais (Figura 4). Figura 3: Páginas 16 e 17 do Diario Segreto di Pollicino (LECHERMEIER, 2010)
Figura 4: Página 23 do Rébecca Dautremer: Artbook (DAUTREMER, 2009) Figura 3: Páginas 16 e 17 do Diario Segreto di Pollicino (LECHERMEIER, 2010)
Figura 4: Página 23 do Rébecca Dautremer: Artbook (DAUTREMER, 2009) Na arte da Pós-Produção, além da utilização do mercado de consumo como matéria-
prima, também a produção artística de outras épocas é incorporada à mixagem em novas
obras. A Pós-Produção não nega a história da arte, ela a reconfigura. "Trata-se de tomar
todos os códigos da cultura, todas as formas concretas da vida cotidiana, todas as obras do
patrimônio mundial e colocá-las em funcionamento (....) saber tomar posse e habitá-las"
(BOURRIAUD, 2009, p.14). Assim, observam-se em obras contemporâneas traços
minimalistas, pop e conceituais, entre outras manifestações. Da mesma forma, é interessante observar que Dautremer, mesmo na escolha de
seus trabalhos, recorre com frequência a referências do passado, tendo ilustrado livros
clássicos da literatura infantil, como o Pequeno Polegar e Alice no País das Maravilhas. Também em suas ilustrações Dautremer deixa transparecer a referência a estéticas de
diferentes períodos artísticos, ora por meio de imagens surrealistas (Figura 5), ora pela
estética da Pop Arte (Figura 6), ora por colagens (Figura 7), todas participando da
construção de uma mesma obra. Como já comentado, em entrevista a ilustradora revela PALÍNDROMO
Nº
10
/
2013 PALÍNDROMO
Nº
10
/
2013 133 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC usar frequentemente como referências as obras de Veemer (por suas cores, luz e sombra);
e Brueghel (por suas criaturas fantásticas) (DAUTREMER, 2011). PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC Figure 5: Páginas 100 e 101 do Diario Segreto di Pollicino (LECHERMEIER, 2010)
Figure 6: Detalhe da página 45 do Diario Segreto di Pollicino (Ibid.)
Figure 7: Página 51 do Diario Segreto di Pollicino (Ibid.) Figure 5: Páginas 100 e 101 do Diario Segreto di Pollicino (LECHERMEIER, 2010)
Figure 6: Detalhe da página 45 do Diario Segreto di Pollicino (Ibid.)
Figure 7: Página 51 do Diario Segreto di Pollicino (Ibid.) Outra característica, presente nas obras da Pós-Produção, é sua relação ou possível
comparação a uma feira de usados, nas quais materiais e objetos de variadas proveniências
e períodos são reagrupados. Tanto objetos com algum valor histórico, como produtos de
caráter efêmero são reciclados a espera de novas funções. "A velha máquina de costura
pode se tornar uma mesa de cozinha, um objeto publicitário de 1975 ou um enfeite para a
sala" (BOURRIAUD, 2009, p. 27), ou seja, o seu uso não se restringe à finalidade para a
qual foi criada. Desde as propostas de ready-made de Duchamp se observa que "utilizar um
produto, é, as vezes, trair o seu conceito". (Ibid, p. 21). Tais aspectos são observados no trabalho de Dautremer, que parece resgatar
objetos dos mais variados períodos históricos, inserindo-os em novos contextos. É o que se
percebe nas ilustrações das Figuras 8 e 9, imagens que remetem a um tempo passado, com
um certo requinte, como uma fotografia antiga envolvida por renda sob um papel de parede
(Figura 8) e uma molheira de porcelana com detalhes dourados (Figura 9), as quais dividem
espaço com referências atuais e populares, como a imagem de um pictograma de
"masculino" na porta de um banheiro rudimentar (Figura 3). Confirmando seu desapego à
exatidão das referências, permitindo-se as mais variadas misturas, Dautremer conta: "Não
tenho dúvidas em desenhar uma tomada na parede atrás de um personagem com roupas
medievais" (SOBRINO, 2011). PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC PALÍNDROMO
Nº
10
/
2013 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC Figure 8: Páginas 24 e 25 do Diario Segreto di Pollicino (LECHERMEIER, 2010)
Figure 9: Páginas 112 e 113 do Diario Segreto di Pollicino (Ibid.) Figure 8: Páginas 24 e 25 do Diario Segreto di Pollicino (LECHERMEIER, 2010)
Figure 9: Páginas 112 e 113 do Diario Segreto di Pollicino (Ibid.) Tomando como outra característica da "feira de usados" muitas obras na Pós-
Produção assumem a estética da disposição aparentemente caótica de produtos,
relembrando o caos cultural, o excesso de oferta e o consumismo. Essa forma de exposição
não necessariamente assume o caráter de crítica, como observado nos pressupostos da
Arte Conceitual, mas aproxima arte ao cotidiano, como parte de um grande "ecossistema
cultural" (BOURRIAUD, 2009, p. 48). Esse "caos visual e estético" pode ser observado nas
composições das Figuras 10 e 11, nas quais os mais variados estilos de desenho se
misturam a colagens de origens também diferenciadas. A variação parece proposital, bem
como as sobreposições, as irregularidades, os desencaixes, ou mesmo as rasuras e as
manchas no papel, relacionando a história do cotidiano da personagem a uma estética do
diário pessoal. Figure 10: Páginas da guarda inicial do Diario Segreto di Pollicino (LECHERMEIER, 2010)
Figure 11: Páginas 68 e 69 do Diario Segreto di Pollicino (Ibid.) Figure 10: Páginas da guarda inicial do Diario Segreto di Pollicino (LECHERMEIER, 2010)
Figure 11: Páginas 68 e 69 do Diario Segreto di Pollicino (Ibid.) Essa mixagem de várias origens, o aplique, o enxerto, a feira de usados, faz surgir
um outro usuário desse mundo Pós-Produzido: "o cidadão do espaço público internacional",
"universalmente exótico" - podendo assumir diferentes identidades, as quais se manifestam
de forma dinâmica e mutante, como se observa em obras de Rirkrit Tiravanija
(BOURRIAUD, 2009). De certa forma, Dautremer também se utiliza de características do
homem mutante por meio de personagens tão diversificados que impossibilitam identificar PALÍNDROMO
Nº
10
/
2013 135 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC quais seriam suas possíveis origens, o que leva a entender que poderiam ser de qualquer
lugar (Figura 10, 11 e 12). Suas vestimentas também variam de medievais a
contemporâneas, de ocidentais à orientais, de reais a ficcionais. PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC Da mesma forma, os
cenários criados misturam traços de cidades modernas com vilarejos antigos, ora contando
sobre o passado, ora falando do contemporâneo, todos parte de uma mesma história num
tempo presente para os personagens. Figure 12: Páginas 68 e 69 do Diario Segreto di Pollicino (LECHERMEIER, 2010) Figure 12: Páginas 68 e 69 do Diario Segreto di Pollicino (LECHERMEIER, 2010) Retomando a relação entre o real e o imaginário, para Bourriaud a Pós-Produção
permite que a ficção e a realidade misturem-se pacificamente, questionando seus próprios
limites. A ficção expõe a realidade ao passo que a realidade zomba da ficção. Daí a
necessidade de se mostrar a possibilidade de novos enredos ao mundo que já está criado. O artista Pierre Joseph, ao defender o uso de personagens de quadrinhos e contos de fadas
em sua obra, diz que "eles não nos convidam a fugir da realidade; pelo contrário, essas
imagens que fazem a aprendizagem do real" (BOURRIAUD, 2009, p. 77). É nesse encontro
entre a ficção e o real que a intervenção humana se manifesta na tela "Zorro" de Mauricio
Cattelan (1993). A tela, rasgada três vezes em forma de "z", remete ao personagem de
ficção Zorro, criado em 1919 pelo escritor norte-americano Johnston McCulley. Zorro, um
transgressor e justiceiro, sai da ficção e deixa suas marcas no mundo real, marcando sua
existência e mostrando que nenhuma imagem deve ser intocável. Essa transgressão
também é observada na obra Edizioni dell'Obligo (1991) do mesmo artista, obra na qual
livros escolares têm sua "capa e título modificados por crianças, numa espécie de desforra e
gozação contra todos os programas", como num ato de "delinquência" (Ibid., p. 72). Se observarmos a capa do livro Diario Segreto di Pollicino (Figura 13) também lá
encontramos sinais claros de transgressão, numa manifestação de descontentamento do
personagem com os paradigmas do livro. Pollicino não admite que seu Diário seja escrito PALÍNDROMO
Nº
10
/
2013 PALÍNDROMO
Nº
10
/
2013 136 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC por outra pessoa, pois afinal, é o Diário de Pollicino. Assim, o personagem risca o nome dos
autores e coloca sobre eles seu próprio nome. Outro sinal de transgressão pode ser
observado na ilustração rasgada e rasurada da capa. 9 Em uma publicação recente Nikolajeva e Scott (2011) apresentam uma visão ampla de diversos estudos a
cerca do livro ilustrado, utilizando para isso uma abordagem semiótica com especial enfoque na articulação entre
texto e imagem. Na mesma obra as autoras esmiúçam os elementos compositivos e narrativos do livro ilustrado,
propondo tipologias e ferramentas para análises de palavra-imagem. PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC Tal fato poderia não ser admitido em
um livro, principalmente em uma capa; para muitos tal ato poderia significar um verdadeiro
sacrilégio, afinal, livros não devem ser riscados e muito menos rasgados. Seu lugar são as
estantes, protegidos com o máximo cuidado, mesmo que isso signifique não manuseá-los. Entretanto, Dautremer se permite rasgar a capa de seu livro e autoriza seu personagem a
rasurá-la, numa contravenção que mistura o real e o ficcional. Também nas folhas de guarda do livro Dautremer autoriza a interferência e
novamente a transgressão no espaço atribuído à apresentação dos autores, a qual,
geralmente acontece de maneira formal. Nesta obra suas fotografias são rasgadas e
misturadas a desenhos, como que numa brincadeira e zombaria infantil. Acompanham
insetos colados sobre as folhas com fitas adesivas transparentes. Talvez seja novamente
Pollicino quem está brincando com os autores, transformando-os em personagens e
caricaturas do mundo real, enquanto ele assume o papel de autor e afirma sua existência. Assim como nas obras da Pós-Produção a autoria é questionada, misturando-se os autores
a sua obra. "Os artistas da Pós-Produção não estabeleceram uma diferença de natureza
entre seus trabalhos e os trabalhos dos outros, nem entre seus gestos e os gestos dos
observadores." (BOURRIAUD, 2009, p. 51). É assim que Dautremer e Pollicino se
confundem, tornando também o leitor cúmplice de seus aventuras, pois dividem com ele seu
diário secreto. Figure 13: Capa do Diario Segreto di Pollicino (LECHERMEIER, 2010)
Figura 14: Folhas de guarda do Diario Segreto di Pollicino (Ibid.) Figure 13: Capa do Diario Segreto di Pollicino (LECHERMEIER, 2010)
Figura 14: Folhas de guarda do Diario Segreto di Pollicino (Ibid.) Por fim, é perceptível que mesmo uma breve análise das ilustrações pôde despertar
para diversas possibilidades de leituras, em uma "teia de elementos", como mencionada por PALÍNDROMO
Nº
10
/
2013
13 137 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC Ramalho e Oliveira (2005, p. 53) (2005). Muitos outros aspectos destas imagens poderiam
ser também analisados como cores, formas, composição, ritmo visual e articulação entre
texto visual e texto verbal9, entre outros, proporcionando à leitura outras dimensões. Cabe
aos educadores e leitores reconhecer essas possibilidades, não restringindo-se a leitura
superficial das imagens, adentrando a suas diferentes camadas de sentidos. Considerações Finais Esse artigo apresentou uma proposta de leitura de imagens do livro ilustrado
associada à leitura de manifestações artísticas, que pode ser utilizada na escola,
despertando o aluno para o exercício da leitura visual crítica em seus diferentes suportes. Para tanto, foi utilizado como instrumento de leitura o modelo desenvolvido por Ramalho e
Oliveira (2005), aplicado a breve exercício, no qual foram analisadas ilustrações do livro
Diario Segreto di Pollicino (2010), de Rébecca Dautremer, a partir da perspectiva da Arte na
Pós-Produção (Bourriaud, 2009). A partir desse exercício foi possível observar diversas possibilidades de leituras
visuais a serem realizadas em sala da aula de forma a provocar o aluno a alcançar níveis
mais profundos e complexos de significações, contribuindo com sua formação visual. Essa
formação, que deve estar presente na escola, não envolve apenas o livro ilustrado ou as
manifestações artísticas; ela permite que um repertório visual seja elaborado, servindo de
base para tantas outras leituras. Acredita-se que a leitura crítica das imagens pode auxiliar
na formação de cidadãos mais conscientes de suas escolhas, tornando-os mais exigentes
frente as possibilidades do mundo que os cercam. ANDERSEN, Hans Christian. Contos e histórias: Hans Christian Andersen; introdução, seleção,
tradução, notas e apêndice de Renata Maria Parreira Cordeiro. 2. ed. São Paulo: Landy Editora,
2004. ARIÈS, Phillipe. História Social da criança e da família. 2. ed. Rio de Janeiro: J. Zahar, 1981. ARIÈS, Phillipe. História Social da criança e da família. 2. ed. Rio de Janeiro: J. Zahar, 1981.
ANDERSEN, Hans Christian. Contos e histórias: Hans Christian Andersen; introdução, seleção,
tradução, notas e apêndice de Renata Maria Parreira Cordeiro. 2. ed. São Paulo: Landy Editora,
2004 LINDEN, Sophie van der. Para ler o livro ilustrado. São Paulo: Cosac Naify, 2011a. LINDEN, Sophie van der. Para ler o livro ilustrado. Abr. 2011b. Disponível em:
<http://editora.cosacnaify.com.br/ObraEntrevista/11359/89/Para-ler-o-livro-ilustrado.aspx> Acesso
em: 30 nov. 2013. Entrevista. LINS, Guto. Livro Infantil? Projeto gráfico, Metodologia, Subjetividade. 2. ed. São Paulo: Rosari,
2003. DAUTREMER, Rébecca. Rébecca Dautremer: Artbook. Paris: Le Chêne, 2009. DAUTREMER, Rébecca. Rébecca Dautremer: Artbook. Paris: Le Chêne, 2009. RAMALHO e OLIVEIRA, S. R. Leitura de Imagens para a Educação. 1998. 288f. Tese (Doutorado
em Comunicação e Semiótica) - Programa de Estudos Pós-Graduados em Comunicação e Semiótica,
PUC/SP. São Paulo, 1998. _____. Imagem também se lê. São Paulo: Edições Rosari, 2005. Referências ARIÈS, Phillipe. História Social da criança e da família. 2. ed. Rio de Janeiro: J. Zahar, 1981 ANDERSEN, Hans Christian. Contos e histórias: Hans Christian Andersen; introdução, seleção,
tradução, notas e apêndice de Renata Maria Parreira Cordeiro. 2. ed. São Paulo: Landy Editora,
2004. 9 Em uma publicação recente Nikolajeva e Scott (2011) apresentam uma visão ampla de diversos estudos a
cerca do livro ilustrado, utilizando para isso uma abordagem semiótica com especial enfoque na articulação entre
texto e imagem. Na mesma obra as autoras esmiúçam os elementos compositivos e narrativos do livro ilustrado,
propondo tipologias e ferramentas para análises de palavra-imagem. PALÍNDROMO
Nº
10
/
2013 138 PALÍNDROMO
Nº
10/2013
–
Programa
de
Pós-‐Graduação
em
Artes
Visuais
–
CEART/UDESC BALDASSARRO, R. Wolf. Banned Books Awareness: Where the Wild Things Are by Maurice
Sendak. 2011. Disponível em: <http://bannedbooks.world.edu/2011/04/10/banned-books-awareness-
wild-maurice-sendak/> Acesso em 30 nov. 2013. BOURRIAUD, Nicolas. Pós-Produção: como a arte reprograma o mundo contemporâneo. São
Paulo: Martins, 2009. BALDASSARRO, R. Wolf. Banned Books Awareness: Where the Wild Things Are by Maurice
Sendak. 2011. Disponível em: <http://bannedbooks.world.edu/2011/04/10/banned-books-awareness-
wild-maurice-sendak/> Acesso em 30 nov. 2013. BOURRIAUD, Nicolas. Pós-Produção: como a arte reprograma o mundo contemporâneo. São
Paulo: Martins, 2009. CADILHAC, Julie. Entrevista DAUTREMER, Rébecca. Rébecca Dautremer: the perfection of line and
illustration. BSC News. Entrevista. Feb. 17 de 2010. Disponível
em:<http://bscnews.fr/20100217768/Bavardages/rebecca-dautremer-la-perfection-du-trait-et-de-
lillustration.html> Acesso em 30 nov. 2013. CALVINO, Italo. Seis propostas para o próximo milênio: lições americanas. São Paulo: Cia das
Letras, 1990. CALVINO, Italo. Seis propostas para o próximo milênio: lições americanas. São Paulo: Cia das
Letras, 1990. DAUTREMER, Rébecca. Rébecca Dautremer: uma ilustradora excepcional. Revista Emília Online. out. 2011. Disponível em:< http://www.revistaemilia.com.br/mostra.php?id=64> Acesso em 20 jul. 2013. Entrevista. DAUTREMER, Rébecca. Melancolia na tinta de Rébecca Dautremer. Revista Continente online. Artes Visuais. mar. 2013. Disponível em: <
http://www.revistacontinente.com.br/index.php/component/content/article/54-artes-visuais/7978-
rebecca-dautremer.html> Acesso em 20 jul. 2013. Entrevista. LECHERMEIER, Philippe; DAUTREMER, Rébecca (ilustração). Diario segreto di Pollicino. Milano:
Rizzoli, 2010. LINDEN, Sophie van der. Para ler o livro ilustrado. São Paulo: Cosac Naify, 2011a. _____. Imagem também se lê. São Paulo: Edições Rosari, 2005. SCOTT, Carole; NIKOLAJEVA, Maria. Livro ilustrado: palavras e imagens. São Paulo: Cosac
Naify, 2011. PALÍNDROMO
Nº
10
/
2013
13
ZIMMERMANN, A., RAMALHO e OLIVEIRA, S. R. E. Ilustrações de livros infantis no ensino de artes. In: XVI Encontro da Associação Nacional de Pesquisadores em Artes Plásticas, 2007. I
Congresso Educação, Arte e Cultura, 2007. ZIMMERMANN, Anelise. As ilustrações de livros infantis: o ilustrador, a criança e a
cultura. 2008. 148 p. Dissertação (Mestrado em Artes Visuais) - Programa de Pós-Graduação em
Artes Visuais, CEART/UDESC. Florianópolis, 2008. ZIMMERMANN, A., RAMALHO e OLIVEIRA, S. R. E. Ilustrações de livros infantis no ensino de artes. In: XVI Encontro da Associação Nacional de Pesquisadores em Artes Plásticas, 2007. I
Congresso Educação, Arte e Cultura, 2007. ZIMMERMANN, Anelise. As ilustrações de livros infantis: o ilustrador, a criança e a
cultura. 2008. 148 p. Dissertação (Mestrado em Artes Visuais) - Programa de Pós-Graduação em
Artes Visuais, CEART/UDESC. Florianópolis, 2008. PALÍNDROMO
Nº
10
/
2013
139
|
https://openalex.org/W4243018846
|
https://www.e-compos.org.br/e-compos/article/download/1026/815
|
Portuguese
| null |
Comunicando diferenças: os processos de hibridização a partir da leitura de la différance nos Estudos Culturais
|
E- compós
| 2,015
|
cc-by
| 9,191
|
www.e-compos.org.br
| E-ISSN 1808-2599 | www.e-compos.org.br
| E-ISSN 1808-2599 | Regiane Regina Ribeiro e Anderson Lopes da Silva Regiane Regina Ribeiro e Anderson Lopes da Silva Resumo O artigo discute o processo de hibridização e as múltiplas
leituras do que é ser ou tornar-se híbrido. A abordagem
inicia-se por Jacques Derrida, com a discussão a respeito
da natureza dividida do signo, la différance, e continua
pelos entendimentos consensuais (e alguns dissensos)
para teóricos como Stuart Hall, Homi Bhabha, Martín-
Barbero e García Canclini, localizando, sempre que
possível, as reflexões no contexto da América Latina. Conclui-se que a hibridização é potencialmente criativa e
ao mesmo tempo conturbada, não se limita às fronteiras
geográficas, linguísticas ou de outra ordem, e é fluida,
desestabilizadora, ambígua e quase sempre contraditória. Palavras-Chave
Hibridização. Estudos Culturais. Comunicação das
diferenças. La différance. O artigo discute o processo de hibridização e as múltiplas
leituras do que é ser ou tornar-se híbrido. A abordagem
inicia-se por Jacques Derrida, com a discussão a respeito
da natureza dividida do signo, la différance, e continua
pelos entendimentos consensuais (e alguns dissensos)
para teóricos como Stuart Hall, Homi Bhabha, Martín-
Barbero e García Canclini, localizando, sempre que
possível, as reflexões no contexto da América Latina. Conclui-se que a hibridização é potencialmente criativa e
ao mesmo tempo conturbada, não se limita às fronteiras
geográficas, linguísticas ou de outra ordem, e é fluida,
desestabilizadora, ambígua e quase sempre contraditória. Palavras-Chave Pensar a hibridização é pensar a mistura e a
mestiçagem como características intrínsecas
aos processos de comunicação. Em um ambiente
deslocalizado, no qual a ausência de bases
fixas e geográficas gera fortes características
de glocalização, fragmentação, individualidade,
liquidez de relações, os processos de hibridização
também produzem, de forma constante, a dúvida
e o questionamento acerca da construção híbrida
das identidades e dos sujeitos pós-modernos. A
comunicação e a cultura, entendidas de modo
ousadamente amplo, encontram-se também nessa
trama de relações híbridas. cia a produtividade e o poder inovador de muitas
mestiçagens interculturais (GARCÍA CANCLINI,
2003, p. 4, tradução nossa). cia a produtividade e o poder inovador de muitas
mestiçagens interculturais (GARCÍA CANCLINI,
2003, p. 4, tradução nossa). cia a produtividade e o poder inovador de muitas
mestiçagens interculturais (GARCÍA CANCLINI,
2003, p. 4, tradução nossa). não se caracterizam mais apenas por elencarem
esta ou aquela característica advinda de seus
elementos originários. O terceiro elemento criado
pela mistura de dois outros possui aspectos que o
tornam singular, híbrido e diferente e não apenas
uma “antonomásia” por excelência. não se caracterizam mais apenas por elencarem
esta ou aquela característica advinda de seus
elementos originários. O terceiro elemento criado
pela mistura de dois outros possui aspectos que o
tornam singular, híbrido e diferente e não apenas
uma “antonomásia” por excelência. Assim, a abordagem inicia-se por Jacques Derrida
com a discussão a respeito da natureza dividida
do signo, la différance. Cabe aqui uma ressalva
de que, mesmo J. Derrida não fazendo parte dos
pensadores fundantes dos Estudos Culturais,
grande parte deles o leu, e suas ideias – com
destaque para a compreensão de la différance –
estão explicitamente citadas nas obras de Hall e
Bhabha, por exemplo. Por isso, a proposta aqui
ensejada também passa pelos entendimentos
da hibridização para Stuart Hall, Homi Bhabha,
Martín-Barbero e García Canclini. Com o objetivo de aprofundar tal discussão, este
artigo traz autores e ideias que desenvolvem a
compreensão de hibridização cultural com base
em confrontos, alguns entendimentos mútuos e
experiências que traduzem bem o quão múltiplas
são as leituras feitas com relação ao que é ser ou
tornar-se híbrido. Como afirma García Canclini: A construção linguística (Bakhtin, Bhabha) e so-
cial (Friedman, Hall, Papastergiadis) do conceito
de hibridização colaborou para que a discussão
saísse dos discursos biologicistas e essencialis-
tas da identidade, da autenticidade e da pureza
cultural. Assim como a mestiçagem contrabalan-
ceou as obsessões por manter incontaminado o
sangue ou as raças no século XIX e em várias
etapas do século XX, a hibridização aparece hoje
como o conceito que permite leituras abertas e
plurais das misturas históricas, além de cons-
truir projetos de convivência despojados das
tendências a “resolver” conflitos multidimensio-
nais através de políticas de purificação étnica. [A
hibridização] Contribui para identificar e explicar
as múltiplas alianças fecundas: por exemplo, do
imaginário pré-colombiano com o novo-hispâ-
nico dos colonizadores e logo com o imaginário
das indústrias culturais (Bernand, Gruzinski), da
estética popular com a dos turistas (De Grandis),
das culturas étnicas nacionais com as das me-
trópoles (Bhabha), e com as instituições globais
(Harvey). Os poucos fragmentos escritos de uma
história das hibridizações colocaram em evidên- Revista
Trabalho apresentado ao Grupo de Trabalho Comunicação e Cultura do XXIII Encontro Anual da Compós, na Universidade
Federal do Pará, Belém, de 27 a 30 de maio de 2014. Palavras-Chave Hibridização. Estudos Culturais. Comunicação das
diferenças. La différance. Revista da Associação Nacional dos Programas de Pós-
Entretanto, mais do que lidar com fusões,
acomodações, crioulizações, sincretismos,
traduções e adaptações híbridas, pensar a
hibridização é também pensar em seus resultados,
suas consequências e seus impactos. Em outras
palavras, da mesma maneira que os elementos
primários que cocriam a hibridização são distintos
entre si (como exigência per si para que ela
ocorra), os elementos derivados de tal mistura já Regiane Regina Ribeiro | regianeribeiro5@gmail.com
Doutora e Mestra em Comunicação e Semiótica pela Pontifícia
Universidade Católica de São Paulo – PUC-SP. Docente permanente
do Programa de Pós-Graduação em Comunicação da Universidade
Federal do Paraná – PPGCOM/UFPR. Anderson Lopes da Silva | anderlopps@gmail.com
Mestre em Comunicação pelo Programa de Pós-Graduação em
Comunicação da Universidade Federal do Paraná – PPGCOM/UFPR. Jornalista, Especialista em Comunicação, Cultura e Arte – PUC-PR e
Membro do NEFICS (Núcleo de Estudos em Ficção Seriada) – UFPR/CNPq. www.e-compos.org.br
| E-ISSN 1808-2599 | 2 Os Estudos Culturais como
rede de estudos transdisciplinares sta da Associação Nacional dos Programas de Pós-Graduação em Comunicação | E-com
Definir os Estudos Culturais como uma disciplina
e com objetos extremamente delimitados de
pesquisa seria negar a essência de sua origem
e de seu desenvolvimento. Entendê-lo como
uma interdisciplina e até mesmo como uma
antidisciplina também não se aproxima de
sua compreensão plena. Essas dificuldades
de classificação dos Estudos Culturais são
motivadas, em sua maioria, pelo amplo espectro
de interesses e de temas de seus pesquisadores. Quem afirma isso é García Canclini, que,
ao falar do assunto, utiliza-se da metáfora
da urbanização e das criações de ruas para
tura do XXIII Encontro Anual da Compós na Universidade campo ou outro, como sustenta García Canclini
(2004, p. 84, 119-121). campo ou outro, como sustenta García Canclini
(2004, p. 84, 119-121). exemplificar tais motivações do surgimento dos
Estudos Culturais: Estruturar campos disciplinares foi, do século
XVIII ao XX, como traçar ruas e organizar terri-
tórios autônomos num tempo em que havia a
necessidade de se defender a especificidade
de cada saber frente às totalizações teológicas
e filosóficas. Mas as disciplinas se entusias-
maram com esta tarefa urbana e, por razões
de segurança, começaram a fechar ruas e a
impedir que elas servissem para aquilo que
originalmente haviam sido construídas: pro-
mover a fácil circulação e o transitar de um
bairro ao outro. Os estudos culturais são tenta-
tivas de reabertura de avenidas ou passagens
para impedir que estas se tornem ampliações
privadas de umas poucas casas (GARCÍA CAN-
CLINI, 2004, p. 122, tradução nossa). Cronologicamente, a origem dos Estudos
Culturais manifesta-se na década de 1950,
na Inglaterra, e, de maneira organizada,
apenas em 1964 passa a “existir no escopo da
Academia” com a criação do CCCS (Centre
for Contemporary Cultural Studies), no
departamento de Inglês da Universidade de
Birmingham. Como apregoa Ana Carolina
Escosteguy, é pela influência de três textos
fundamentais que o movimento ganha corpo. Isto é, com The Uses of Literacy (1957), de
Richard Hoggart, Culture and Society (1958), de
Raymond Williams, e The Making of the English
Working-class (1963), de E. P. Thompson, os
Estudos Culturais passam a jogar novas luzes
sobre as conceituações de cultura e suas inter-
relações com o poder, a economia e a história
“dos de baixo” (ESCOSTEGUY, 2010, p. 27-28). Com relação a essa condição de subordinação
e exclusão, García Canclini defende que
o afastamento dos eixos tradicionais da
identidade em um cenário de crescimento
e produção global da cultura não pode ser
considerado como situação desagradável ou
temível, porque vivemos: comunicação e da cultura – tal qual já o faziam
os britânicos –, os latino-americanos refletem
suas ideias em um cenário no qual os conceitos
de nacionalismo, populismo, resistência,
anarquismo, apropriação e ressemantização
eram vivenciados tanto na vida do analista
quanto no decorrer de suas análises. no plano das representações e no imaginário
social e representaram uma forma violenta
de negação da alteridade. Disso emergiram
sujeitos que foram massacrados, que souberam
resistir e que continuam afirmando suas
identidades na sociedade – mesmo obedecendo
a relações assimétricas, de subordinação e de
acentuada exclusão. E tudo isso em um momento em que a matriz
de pensamento das Ciências Sociais estava
cristalizada sob a égide de um marxismo
a-historicizado e de uma visão cristã como
condições sine qua non para se pensar a
comunicação e a cultura nas pesquisas (KUNSCH,
2002, p. 14). Com uma proposta diametralmente
oposta, os Estudos Culturais na América Latina
começam a questionar os liames e as zonas
transfronteiriças da cultura popular e das
indústrias culturais, tentando localizar outros
aspectos que não apenas os buscados pelos
trabalhos de linha marxista-cristã2. Revista da Associação Nacional dos Programas de Pós-Graduação em Comunicação | E-compós, Brasília, v.18, n.1, jan./abr. 2015. 4/18
om relação a essa condição de subordinação
exclusão, García Canclini defende que
afastamento dos eixos tradicionais da
dentidade em um cenário de crescimento
produção global da cultura não pode ser
onsiderado como situação desagradável ou
emível, porque vivemos:
um tempo de fraturas e heterogeneidade, de
segmentação dentro de cada nação e de comu-
nicações fluidas com as ordens transnacionais
da informação, da moda e do saber. Em meio a
esta heterogeneidade encontramos códigos que
nos unificam, ou que ao menos permitem que
nos entendamos (GARCÍA CANCLINI, 1999, p. 85). egundo Pinheiro (1995, p. 16), os signos na
mérica Latina apresentam uma mobilidade
muito grande, o que não significa que seja
iferente em outros lugares do mundo, mas
qui essa mobilidade chega a ser desmesurada. sso se deve à mestiçagem e ao choque dos
rocessos civilizatórios que fazem os signos
de Jesús Martín-Barbero, Néstor García Canclini e,
s, outros nomes possuem relevância, como o de Carlos
Sunkel, José Joaquín Bruner (Chile); Renato Ortiz (Brasil);
COSTEGUY, 2010, p. 47). 2 Os Estudos Culturais como
rede de estudos transdisciplinares Vistos mais como um movimento ou uma
rede de estudos (JOHNSON, 2004) do que um
campo delimitado e com bases epistemológicas
e metodológicas muito restritas, os Estudos
Culturais podem ser entendidos como o campo
no qual a Comunicação Social, a Sociologia,
a Antropologia, a História, a Linguística e
os Estudos Literários ganham um olhar que
não se contenta com um ou com outro objeto
de suas áreas específicas. Pelo contrário, na
origem dos Estudos Culturais as leituras do
mundo, de seus objetos e de seus sujeitos
foram feitas pela lente de pesquisadores que
provinham de distintas áreas. Áreas que,
ao negarem uma ortodoxia científica rígida
demais às pesquisas, se complementavam. Assim, o uso do termo “transdisciplina” se
apresenta adequado, uma vez que a visão
dessa rede de estudos não se restringe a um Revista da Associação Nacional dos Programas de Pós-Graduação em Co
Já no contexto da América Latina, a mera
tradução de “Cultural Studies” para Estudos
Culturais não representa uma transposição
das ideias do grupo britânico para o grupo dos
latinos. Com especificidades que caracterizam o
movimento no continente, os Estudos Culturais
começam a florescer na América Latina durante
a década de 1980, valorizando o receptor e sua
capacidade tanto de resistir como de responder
(visualizando-o como cocriador de mensagens
e de sentidos). Entretanto, mais do que
seguir a visão de um receptor como sujeito da 2 Segundo Ecosteguy, destacam-se nessa perspectiva as reflexões de Jesús Martín-Barbero, Néstor García Canclini e,
posteriormente, Guillermo Orozco Gómez. Além desses três autores, outros nomes possuem relevância, como o de Carlos
Monsiváis, Jorge González, Rossana Reguillo (México); Guillermo Sunkel, José Joaquín Bruner (Chile); Renato Ortiz (Brasil);
Beatriz Sarlo, Aníbal Ford (Argentina) e Rosa Maria Alfaro (Peru). (ECOSTEGUY, 2010, p. 47). apresentarem tal característica. Dessa forma,
parece ficar mais clara a ideia de Pinheiro quando
afirma que se pensa de modo diferente nesse
continente e que não seria possível comportar-
se de outra maneira enquanto descendentes de
uma atitude epistemológica diferente, pois a
mente trabalha os signos, mais através da fricção
de superabundâncias alógenas do que pelos
mecanismos binários de inclusão e de exclusão. configurações de forças. Novas saídas do campo
e diásporas se desenharam, forçando limites e
reconfigurando espaços. Esse processo acarretou
mudanças no interior do repertório cultural dos
povos, e, consequentemente, na ideia tão fortemente
demarcada de “fronteiras culturais” (como é possível
também visualizar na América Latina). configurações de forças. Novas saídas do campo
e diásporas se desenharam, forçando limites e
reconfigurando espaços. Esse processo acarretou
mudanças no interior do repertório cultural dos
povos, e, consequentemente, na ideia tão fortemente
demarcada de “fronteiras culturais” (como é possível
também visualizar na América Latina). dos Programas de Pós-Graduação em Comunicação | E-compós, Brasília, v.18, n.1, jan./abr. 2015. 5/18
Ou seja, os processos migratórios e a
desterritorialização desconstituem o conceito de
comunidade nacional e dificultam a localização
linear de repertório de costumes, mitos e práticas
que formavam um universo simbólico e, em
consequência, configuravam as identidades. Tais repertórios tornaram-se transculturais,
implodindo o conceito de fronteiras delimitadas,
que não têm correspondência exclusiva com os
territórios em que se encontram e tampouco
apresentam com estes uma relação de oposição. Nesse contexto, García Canclini (2003) diz que
a ideia de hibridização cultural está colocada já
na transposição das fronteiras e será percebida
pelo processo transcultural, que caracteriza a
expressão das culturas. Isso determinará uma
construção identitária sempre em trânsito, na
qual a cultura desterritorializada buscará seu
“não lugar”, ocupando de maneira incisiva um
novo locus de expressão. Isso significa que os processos de produção
de sentido nesse continente apresentam uma
diversidade tão exacerbada que tendem a romper
a lógica binária e subverter uma valoração
simplificadora que exclui o raciocínio e produz
uma reação de conforto rápido e cômodo em
um mundo de complexidade crescente. Baggio
apresenta essa dicotomia como algo: [...] do pensamento tradicional, com forte in-
fluência do pensamento grego que foi manti-
do no decorrer da história e que ainda se faz
presente na sociedade contemporânea, é a
criação de linguagem dicotômica pela qual o
humano é percebido, tratado, educado. 3 A questão multicultural:
desafios e perspectivas Pensar a questão multicultural na América Latina
requer um olhar sobre como as relações
interétnicas fortes e constantes construíram
seus espaços. A escravização e a eliminação
física do “outro” estiveram sempre presentes 2 Segundo Ecosteguy, destacam-se nessa perspectiva as reflexões de Jesús Martín-Barbero, Néstor García Canclini e,
posteriormente, Guillermo Orozco Gómez. Além desses três autores, outros nomes possuem relevância, como o de Carlos
Monsiváis, Jorge González, Rossana Reguillo (México); Guillermo Sunkel, José Joaquín Bruner (Chile); Renato Ortiz (Brasil);
Beatriz Sarlo, Aníbal Ford (Argentina) e Rosa Maria Alfaro (Peru). (ECOSTEGUY, 2010, p. 47). de retornar ao lugar de origem, os espaços de
exploração e a articulação das identidades
híbridas devem construir-se com base nas
discussões sobre diversidade, as quais são
produzidas dentro dos espaços de significação
e das estruturas de poder, problematizando-as
e provocando o questionamento em torno da
experimentação de identidades alternativas. só a escrita não dá conta de representar a fala
de forma isonômica e vice-versa”, como também
radicaliza “ainda mais os pressupostos e as
consequências do conceito de representação, tão
caro à filosofia da identidade que predomina no
Ocidente há séculos, desde Platão e Aristóteles até
Saussure e Lévi-Strauss” (BARBOSA, 2012, p. 119). Salientando que la différance não é nem
uma palavra e nem um conceito, o filósofo
retoma os dois sentidos do verbo diferir (verbo
latino differre): o de diferir na temporização
(“temporizar é recorrer, consciente ou
inconscientemente, à mediação temporal e
temporizada de um desvio que suspende a
consumação e a satisfação do desejo ou da
vontade, realizando-o de fato de um modo que
lhe anula ou modera o efeito”) e de diferir com
o sentido de não ser idêntico (dos aspectos
relacionados à “alteridade de dissemelhança”)
(DERRIDA, 1991, p. 38-39). Esse
paradigma antropológico opõe natureza à cul-
tura, fragmenta o humano em razão-emoção,
sujeito-objeto,
academicismo-objetividade,
corpo-alma, matéria-espírito, masculino-femi-
nino, hemisférios cerebrais direito-esquerdo e
o conhecimento a partir de identidade e da não
identidade. (1999, p. 26) Revista da Associação Naciona
Assim, na América Latina, a fronteira se configura
como lugar onde uma cultura se expressa trazendo
em si a marca de uma relação desigual, na qual
a violência é exercida no confronto com o outro,
com a alteridade. E, quando não existe maneira Essa atitude epistemológica sui generis se
intensifica na segunda metade do século XX,
quando se testemunha um grande deslocamento
populacional incentivado pelas dinâmicas
estabelecidas com os novos reagrupamentos e Tal jogo de estratégia tem por norte a dualidade
ausência/presença. E, desse modo, consegue
apresentar a natureza dividida do signo, isto é,
la différance possui o duplo significado de se
sobrepor e, ao mesmo tempo, ser distinto. John
Storey, em “Teoría Cultural y Popular”, comenta
que Derrida, mesmo levando em consideração o
sistema de diferenças como a localização que faz
com que os signos produzam sentido (noção
proposta por Saussure), acrescenta a ele a ideia
de que um “significado sempre é adiado, nunca
está completamente presente, sempre presente
e ausente”. E dá como exemplo o dicionário:
“se buscamos o significado de uma palavra
em um dicionário, encontramos um contínuo
adiamento do significado”, explica Storey (2001,
p. 123, tradução nossa). Um jogo infinito de
reenvios de sentido e de signos que se remetem
na presença/ausência. e “diferante” de diferenças, como coloca Derrida
(1991, p. 43), é uma não origem, uma vez que,
por não ser um fundamento ou um ponto de
partida simplista, de igual forma não possui um
fim ou ponto de chegada: é uma “estratégia sem
finalidade” (DERRIDA, 1991, p. 38). Sobre o assunto, Derrida afirma que la différance
é o que faz com que o movimento da significação
só se torne possível por cada elemento dito
“presente”, ou seja, aquele que aparece sobre a
cena da presença possa se relacionar com outra
coisa que não ele próprio (DERRIDA, 1991, p. 45). Dessa forma, prossegue ele, cada elemento do
presente, guardando em si a marca do elemento
passado, também se deixa “moldar”, refazer-se por
sua relação com o elemento futuro. Assim: É necessário que um intervalo o separe do que
não é ele para que ele seja ele mesmo, mas
esse intervalo que o constitui em presente deve,
no mesmo lance, dividir o presente em si mes-
mo, cindindo, assim, como o presente, tudo o
que a partir dele se pode pensar, ou seja, todo
o ente na nossa língua metafísica, particular-
mente a substância e o sujeito: Esse intervalo
constituindo-se, dividindo-se dinamicamente,
é aquilo a que podemos chamar espaçamento,
devir-espaço do tempo ou devir-tempo do espa-
ço (temporização) (DERRIDA, 1991, p. 45). o filósofo de la différance A definição do sentido de la différance torna-se
complicada já no momento de uma possível
tradução do termo à língua portuguesa. Isso
porque, em francês, o vocábulo produz uma
dupla significação por tratar-se de um neografismo
homófono à palavra différence (diferença, em
português). A troca de um a por um e, como se
poderia esperar, não é uma simples mudança sem
consequências maiores. Quando Jacques Derrida
(1930-2004) proferiu sua palestra “La Différance”,
na Sociedade Francesa de Filosofia, em 27 de
janeiro de 1968, tal “falha silenciosa à ortografia”
foi muito significativa, ou seja, a troca do
“legítimo” e pelo “transgressor” a, como colocam
os tradutores de “Margens da Filosofia” (1991),
trouxe uma crítica à tradição filosófica ocidental,
de modo sutil, que deixava explícito um exemplo
de seus sintomas: o fonocentrismo, ou seja, o
privilégio da fala sobre a escrita. Revista da Associação Nacional dos Programas de Pós-Graduação em Co
Pós-estruturalista, o francês Jacques Derrida,
filósofo da desconstrução, apresenta la différance
como algo que remete ou reenvia para dois
movimentos distintos, a saber: 1) a diferenciação
como a produção de diferenças, alteridades, das
não identidades em um sistema sígnico – pondo
em xeque pressupostos da linguística de Saussure;
e 2) o espaçamento, o desvio, a temporização,
o retardamento, o cálculo sucinto “que faz com
que um sentido seja sempre antecipado ou
restabelecido em posteridade” (BENNINGTON;
DERRIDA, 1996, p. 58). Essa “origem” estruturada O filósofo Lázaro Barbosa destaca que la
différance mostra, dessa maneira “[...] que não 5 A leitura de la différance
por um viés culturalista Mas, afinal, qual a leitura que os Estudos
Culturais fazem da obra de Derrida? Qual
ressignificação é dada pelos teóricos da cultur
acerca de la différance? Continuando no tom
ensaístico que constitui este trabalho, é possív
responder afirmando que Hall e Bhabha são os
dois autores que mais frequentemente fazem u
da desconstrução e do vocabulário derrideano Dessa forma, la différance seria “o movimento
do jogo que produz as diferenças, os efeitos de
diferença”, uma estratégia. Em outras palavras:
la différance “não é mais simplesmente um
conceito, mas a possibilidade de conceitualidade,
do processo e do sistema conceitual em geral”
(SANTIAGO, 1976, p. 23-24). O exemplo das utilizações da obra de Derrida
em Hall é bem mais nítido do que nos outros levado a praticar a desconstrução como modo
crítico de abordar o “real” e o “autêntico”, isto
é, a crítica ao que se toma por realidade e sua
demasiada busca por um discurso científico
limpo de qualquer traço subjetivo. Outro uso de
Derrida por parte de Bhabha é descrito por Souza
pela correlação conceitual. É interessante notar
que as duas conotações de diferir descritas pelo
filósofo francês também são utilizadas pelo crítico
indiano. O conceito de imagem em Bhabha, por
exemplo, é lido como a ‘economia do suplemento’
para Derrida, e é tido como “perigoso”: porque
a imagem em si, como ponto de identidade, é
ambivalente, pois é representação e signo, ou
seja, é sempre “fendida”, tanto espacialmente
(torna presente algo que está ausente) quanto
temporalmente (representa algo que veio antes e,
por isso, torna-se uma repetição) (SOUZA, 2004). autores. Para se ter uma ideia, Hall recusa-se
a pensar a identidade cultural dos povos do
Caribe em concepções binárias ou até mesmo
excludentes: o autor passa a visualizar la
différance como a saída para suas explicações. A compreensão da pós-colonialização em Hall
baseia-se em uma “releitura” de Derrida, uma
vez que ela é entendida como um processo global
fundamentalmente transnacional e transcultural. Lázaro Barbosa comenta que tal “ruptura indica
uma nova leitura das identidades culturais que se
pretendem autônomas e autoproduzidas, pelo jogo
da différance, no qual colonizadores e colonizados
se inscrevem um através do discurso do outro” e,
como consequência dessa ambivalência, eles se
apropriam “mutuamente em suas semelhanças e
diferenças” (BARBOSA, 2012, p. 120). As diferenças que constituem a identidade
cultural caribenha ocorrem por meio de “places
de passage, e significados que são posicionais
e relacionais, sempre em deslize ao longo de
um espectro sem começo nem fim”, explica
Hall (2003, p. 33). E, de maneira mais explícita:
“Naturalmente, o que faço aqui é traduzir da
filosofia à cultura e expandir o conceito de
Derrida sem autorização – embora, espero, não o
faça contra o espírito de seu sentido/propósito”
(HALL, 2003, p. 92). Já Bhabha, ao criticar
algumas metodologias de análise do discurso do
colonizador sobre o colonizado, recorre a Jacques
Derrida pela ideia de não separar aquele que
pesquisa daquilo/daquele que é pesquisado. diferença por meio da qual estabelecem uma
linha de jogo entre duas forças: uma que
empurra e outra que puxa. factuais”. Entretanto, tanto ele como Derrida,
quando lançam mão de la différance em suas
obras, de forma alguma estão “a-historicizando”
seus debates, tendo em vista que “toda vez
que se atribui uma validade intrínseca a algo,
transcendendo limites de tempo e espaço, é
necessário incansavelmente contextualizar e
historicizar tal atribuição”. Mais do que isso:
é preciso “desuniversalizar” e “mostrar” tais
questões como produtos de suas condições
históricas, mas também culturais e ideológicas
produtivas, finaliza Souza (2004). Desse modo, o conceito de fronteiras – como
muro e defesa – sofre uma alteração, passando-
se a considerá-las pontos de contato, espaços de
relações que se (re)organizam pela intervenção
dos sujeitos através delas. O redimensionamento
desses pontos/superfícies de contato, nas
relações, é consequência das mudanças rápidas e
progressivas do conceito tradicional de fronteiras,
por intermédio das quais os indivíduos projetam-
se no corporal e no imaginário. Seu
entendimento, com base na obra derrideana, é Homi Bhabha não apenas faz uso direto de
Derrida, como também o “defende” ao rejeitar
a crítica que é apontada ao filósofo e, por
conseguinte, ao próprio trabalho também. A c
diz respeito ao a-historicismo que, supostame
estaria arraigado nas suas discussões acerca
indeterminação e da ambivalência: tal valoriz
seria desnecessária e irresponsável em relaçã
referentes históricos que rodeiam os seus deb Revista da Associação Nacion
Souza diz que, ao sair em defesa de Derrida e
enfatizar a hibridização que permeia a linguagem,
Homi Bhabha procura mostrar que é impossível
lidar com tais questões tentando caracterizar a
linguagem e seus usos com “valores objetivos e 3 Especificamente porque até mesmo García Canclini, um dos grandes nomes na discussão da hibridização cultural, fala que o
objeto de seu livro não é a “hibridez, e, sim, os processos de hibridação” (GARCÍA CANCLNI, 2011, p. XXVII). Logo, a discussão
centra-se neste artigo não na busca de um tensionamento conceitual que focalize a hibridização em si, mas, de modo
correlato, uma busca que traga os processos hibridizadores sob o foco da comunicação e sua interface com outros campos
(como a cultura). 6 Qual é o espaço da
diferença na comunicação? A contribuição dessa reflexão (para além do
que os autores aqui utilizados já discutem)
está não na problematização do conceito3 do
ponto de vista comunicacional e intercultural. É dizer que: somente a partir do tensionamento
conceitual entre aquilo que é abordado por
García Canclini (2011), por exemplo, e os
objetos empíricos estudados pelo campo da
comunicação4 é que os processos podem ser
correlacionados entre teoria e prática, entre
plano conceitual e plano praxiológico. Em
outras palavras, a conceituação de hibridização
cultural encontra-se no plano das ideias e das
reflexões defendidas pelas múltiplas visões As mídias como processos culturais e
comunicacionais criam seus moldes e seus
sistemas considerados aqui como elementos
de importância na produção de sentido e (re)
construção social midiatizada. Na atualidade,
apresenta-se paralelamente à globalização
econômica a existência de uma globalização de
práticas sociais, em um contexto atravessado
por diferentes mídias que se entrelaçam
configurando um complexo cenário midiatizado. Nesse conjunto de acontecimentos, ganham
força os movimentos/fluxos de sujeitos pela
experiência da miscigenação cultural, da 3 Especificamente porque até mesmo García Canclini, um dos grandes nomes na discussão da hibridização cultural, fala que o
objeto de seu livro não é a “hibridez, e, sim, os processos de hibridação” (GARCÍA CANCLNI, 2011, p. XXVII). Logo, a discussão
centra-se neste artigo não na busca de um tensionamento conceitual que focalize a hibridização em si, mas, de modo
correlato, uma busca que traga os processos hibridizadores sob o foco da comunicação e sua interface com outros campos
(como a cultura). 3 Especificamente porque até mesmo García Canclini, um dos grandes nomes na discussão da hibridização cultural, fala que o
objeto de seu livro não é a “hibridez, e, sim, os processos de hibridação” (GARCÍA CANCLNI, 2011, p. XXVII). Logo, a discussão
centra-se neste artigo não na busca de um tensionamento conceitual que focalize a hibridização em si, mas, de modo
correlato, uma busca que traga os processos hibridizadores sob o foco da comunicação e sua interface com outros campos
(como a cultura). 4 Como as telenovelas, processos de hibridização aliados aos sistemas culturais, a discussão das mediações socioculturais
para além dos meios massivos, entre outros objetos. 4 Como as telenovelas, processos de hibridização aliados aos sistemas culturais, a discussão das mediações socioculturais
para além dos meios massivos, entre outros objetos. 6 Qual é o espaço da
diferença na comunicação? 4 Como as telenovelas, processos de hibridização aliados aos sistemas culturais, a discussão das mediações socioculturais
para além dos meios massivos, entre outros objetos. www.e-compos.org.br
| E-ISSN 1808-2599 | que são de seus antigos países. O destaque da
conceituação de hibridização para Hall encontra-se
no seu entendimento dos processos hibridizadores
como formas potenciais de fontes criativas, isto é,
formas criadoras de novas percepções de mundo. O teórico cultural jamaicano afirma isso dizendo
que “o ‘hibridismo’ e o sincretismo – a fusão entre
diferentes tradições culturais – são uma poderosa
fonte criativa”, que produz inovadoras formas
de cultura, “mais apropriadas à modernidade
tardia que às velhas e contestadas identidades do
passado” (HALL, 2000, p. 91). transcomunicacionais, viabilizadoras dos
processos que impulsionam as culturas híbridas,
as quais, então, liberariam as análises culturais de
seus processos de fundamentalismos identitários
(GARCÍA CANCLINI, 2000). transcomunicacionais, viabilizadoras dos discutidos neste artigo, mas os processos e as
formas de hibridização comentadas por eles
podem ser analisados para além do estudo
seminal antropológico, literário, filosófico,
folclórico e apenas cultural dos autores, ou seja,
eles podem ser lidos na interface da comunicação
e dos estudos culturais, destacando, entre outras
coisas, a especificidade do teor comunicativo de
massa, do consumo cultural e das tensões entre o
antigo e moderno, entre o local e o global, etc. para assumir-se sociocomunicacional, atuando
em redes comunicacionais deslocadas e onde a
diferença marca presença (GARCÍA CANCLINI,
1993, p. 44). Os processos migratórios e a tecnologização
das relações, inseridas em um espaço no
qual convivem simultaneamente todas as
temporalidades históricas, acarretam uma
desarticulação das séries culturais classificadas
como “cultas”, “populares”, e, ainda, “massivas”,
ou, como coloca McCarthy, o “olhar do poder,
suas normas e pressupostos, precisa ser
desconstruído” (1998, p. 156, tradução nossa). A América Latina pode ser vista como o
exemplo mais visível desses novos processos de
produção industrial, eletrônica e informática
que reorganizam o que antes era dividido em
fronteiras do culto e do popular. Martín-Barbero
(2002, p. 146) observa que as indústrias culturais
estão reorganizando não apenas as identidades
subjetivas e coletivas, mas também formas de
diferenciação simbólica. O autor ainda explica
que é justamente pelo estudo sistemático
dessas produções “mestiças” e dos processos
de comunicação massiva que será possível
compreender essas novas demarcações: agora
reorganizadas em uma sociedade também híbrida
e marcada pela diferença. Assim, fronteira tensiona e confronta as
realidades. Essas, por sua vez, são fortalecid
nas discussões raciais, étnicas, de gênero, n
meio ambiente, nas concepções religiosas, n
sexualidade, na política do corpo, na língua,
García Canclini concorda com o fato de que
ser relativizada a noção de identidade. Afirm
os processos de hibridação é esvaziar a ideia
identidade “autêntica”, tal como a concebe u
forte tendência da Antropologia, assim como
alguns enfoques de pesquisadores implicado
com os Estudos Culturais. Dessa maneira, o
autor propõe um deslocamento do objeto de
estudo: da identidade para a heterogeneidad
hibridização interculturais. Isso deu origem ao conceito de “região
transfronteiriça”, segundo o qual coexistem,
em um mesmo grupo, vários códigos simbólicos
que garantem uma identidade multiétnica,
transitória, mutante e migrante, formada por
elementos cruzados de várias culturas. A
definição deixa de ser unicamente socioespacial As grandes cidades, palcos dos processos mais
interessantes de hibridização, apresentam
“fronteiras porosas”, permeáveis às redes 7 A criatividade e o antagonismo
na cultura: os processos
de hibridização em Hall Revista da Associação Nacional dos Programas de Pós-Graduação em Comunicação | E-compós, Brasília, v.18, n.1, jan./abr. 2015. 11/18
o te c at a , que p odu
o ado as o
as
de cultura, “mais apropriadas à modernidade
tardia que às velhas e contestadas identidades do
passado” (HALL, 2000, p. 91). Com base na leitura que Hall faz de Jacques
Derrida, o pesquisador Lázaro Barbosa comenta
que, ao ressaltar toda a dinâmica da hibridização
nos processos multiculturais, diaspóricos e pós-
coloniais, o pesquisador jamaicano pautou sua
busca na “superação de dicotomias envolvendo
os discursos políticos, culturais e acadêmicos”
advindos das tradicionais direita e esquerda. “Em suma, se Hall não pode ser considerado
um teórico derrideano, pelo menos é digno
de atenção pelos argumentos defendidos e
pela disseminação da différance nos estudos
culturais” (BARBOSA, 2012, p. 123). De igual modo, pensar o processo da hibridização,
para o pesquisador, é questionar-se sobre aquilo
que ele chama de “proliferação subalterna
da diferença” (HALL, 2003, p. 60), isto é, a
desestabilização da cultura e as perturbações
sociais que – também vistas na linguagem – são
promovidas pelo embate entre o moderno e o A conceituação de Stuart Hall dada ao hibridismo
confunde-se com sua experiência enquanto reflexo
de um fluxo migratório de caribenhos para a
Inglaterra no século passado. Dito de outro modo,
tal reflexo é projetado em suas obras como o retrato
daquilo que se convencionou chamar de estudos
pós-colonialistas e análises pautadas na diferença,
na desterritorialização e no descolecionamento
de bases tidas como puras e tradicionais. Hall
compreende a hibridização cultural como algo
que “não se refere a indivíduos híbridos, que
podem ser contrastados com os ‘tradicionais’ e
‘modernos’ como sujeitos plenamente formados”. Para ele, o conceito trata-se “de um processo
de tradução cultural, agonístico, uma vez que
nunca se completa, mas que permanece em sua
indecidibilidade” (HALL, 2003, p. 74). Com base na leitura que Hall faz de Jacques
Derrida, o pesquisador Lázaro Barbosa coment
que, ao ressaltar toda a dinâmica da hibridizaç
nos processos multiculturais, diaspóricos e pó
coloniais, o pesquisador jamaicano pautou sua
busca na “superação de dicotomias envolvendo
os discursos políticos, culturais e acadêmicos”
advindos das tradicionais direita e esquerda. “Em suma, se Hall não pode ser considerado
um teórico derrideano, pelo menos é digno
de atenção pelos argumentos defendidos e
pela disseminação da différance nos estudos
culturais” (BARBOSA, 2012, p. 123). antigo, entre o que se entende por culto e não
culto, entre classes superiores e inferiores. suscitadas pelo pós-colonialismo: nesse caso,
tendo a Inglaterra (colonizadora) e a Índia
(colonizada) como o pano de fundo de suas
discussões. Suas ideias e seus conceitos com
relação à hibridização surgem em um contexto
de confronto na literatura de ex-colonizados e ex-
colonizadores; um embate entre qual das formas
de representação literária representaria mais ou
menos fidedignamente a realidade dos nativos. De todos esses apontamentos, há pelo menos uma
certeza postulada por Stuart Hall: entre esses
locais de contato e entre as comunidades e os
sujeitos imaginados (termos tomados de Benedict
Anderson), a diáspora muda os que saem, muda os
que já estão e muda também os que ficaram. Para
entendê-los, é preciso compreender os lugares de
passagem e de diferença, quer dizer, os locais onde
os significados também se tornam relacionais,
posicionais e nunca definitivos. A preocupação com os processos híbridos surge
de sua experiência própria como membro de uma
elite local (os Parsi, de Mumbai), presente em uma
sociedade colonizada pelos ingleses durante dois
séculos. De igual modo, tais discussões iniciam-se
também do objeto de análise de seus primeiros
trabalhos, isto é, o discurso colonial britânico na
Índia do século XIX. Assim, seguindo o pensamento de que a
hibridização é potencialmente criativa e ao
mesmo tempo conturbada, as desestabilizações
da cultura também são potenciais. Pensar nessas
desestabilizações é pôr sob tensão a suposta
neutralidade cultural por parte de um estado liberal,
por exemplo, com o pensamento de uma “cultura
além das culturas” versus os “particularismos”
que se universalizaram de modo hegemônico como
“isto é cultura, isto não é cultura”. É por isso que
Hall aponta a hibridização como o desestabilizador
cultural por excelência: o híbrido não aceita
conceitos fechados nem oposições binárias. Ao criticar a análise de imagens e a análise
ideológica dos textos, Bhabha não traz a mes
abordagem “realista” da literatura e das suas
representações identitárias da cultura e da
hibridização. Ele se utiliza da desconstrução
de Derrida: a crítica de Bhabha está voltada
principalmente à representação violenta que
separa sujeito e objeto, fixando, sempre, o
sujeito em uma posição de inteligibilidade
hierarquicamente privilegiada (SOUZA, 2004 7 A criatividade e o antagonismo
na cultura: os processos
de hibridização em Hall Revista da Associação Nacional dos Progr
De igual modo, pensar o processo da hibridização,
para o pesquisador, é questionar-se sobre aquilo
que ele chama de “proliferação subalterna
da diferença” (HALL, 2003, p. 60), isto é, a
desestabilização da cultura e as perturbações
sociais que – também vistas na linguagem – são
promovidas pelo embate entre o moderno e o E é justamente durante a tradução cultural que
os sujeitos diaspóricos têm diante de si uma
cultura que não as assimila, que não as integra
e, ao mesmo tempo, não perdem completamente
suas identidades originárias: elas ainda guardam,
muitas vezes pela forma oral, características 8 A tradução cultural em Bhabha:
a hibridização vista pelas representações
linguísticas e identitárias Em outras palavras, de acordo com Souza
(2004), é pelo caminho da linguagem que
Homi Bhabha busca a identidade, ou seja, é no
nível do discurso que ele traça sua rota com o Tal qual Stuart Hall, o crítico indiano Homi
Bhabha escreve, pensa e vive as problemáticas aos estereótipos não são considerados na mesma
medida que Bhabha. Diferentemente de García
Canclini, que vê na hibridização um processo
multicultural e capaz de possibilitar o respeito,
a valorização e a tolerância às diversidades
culturais, para Hall e Bhabha isso resulta do
choque, do embate e, por isso, não traz consigo
uma via constante de entendimento, ou seja, a
visão do autor jamaicano e a do autor indiano não
são tão “elogiosas” em relação aos processos de
mestiçagem, de mistura. intuito de compreender as traduções culturais
e a hibridização que as constituem. O enfoque
de Bhabha recai no que estava em jogo, ou
seja, entender “se eram as linguagens usadas
para representar os sujeitos ou [...] a questão
da construção da identidade” que realmente
interessava, explica Souza (2004, p. 114). do entendimento de uma “tradução cultural”, o
crítico indiano menciona que a hibridização é
um modo de conhecimento, um meio de entender
e também de perceber as transformações –
conflitantes – sociais e culturais (BHABHA, 2002
apud SOUZA, 2004, p. 113). Dessa maneira, pensar a hibridização em Bhabha
é partir do conceito de discurso como uma prática
significatória. Em outras palavras, na esfera
linguística a identidade e a sua representação
estão umbilicalmente presentes em um “processo
que postula a significação como uma produção
sistêmica situada dentro de determinados
sistemas e instituições de representação –
ideológicos, históricos, estéticos, políticos” da
tradução cultural (BHABHA, 1984, p. 98). A busca pela identidade, conforme Homi
Bhabha, é sempre agonística porque a
identidade é sempre uma imagem, que continua
sendo imagem, ainda que “muito autêntica”
(por isso, nunca é substancial). Bhabha
explica que o acesso à imagem da identidade
seguindo uma imagem só é possível por meio
de uma negação do sentido de originalidade
ou da ideia de plenitude, “através do princípio
de deslocamento e diferenciação (ausência/
presença; representação/repetição)” que, de
uma maneira ou de outra, torna a realidade
ambígua. E, nesses termos, o processo
relacional da identidade (a imagem) é, de
modo dúbio, uma “substituição metafórica,
uma ilusão de presença” e, por isso mesmo,
uma “fronteira movediça da alteridade na
identidade” (BHABHA, 2010). Assim, quaisquer das tentativas de representação
tornam-se híbridas por conterem traços dos dois
discursos, dos discursos (o do colonizado e do
colonizador) que se misturam em um complexo
jogo de alteridade e distinção, no qual intentar
uma autenticidade é algo impossível. Em uma
definição que antecipa suas discussões acerca ______. Diferentes, desiguales y desconectados:
mapas de la interculturalidad. Gedisa Editorial:
Barcelona, 2004. ______. Diferentes, desiguales y desconectados:
mapas de la interculturalidad. Gedisa Editorial:
Barcelona, 2004. ______. Malestar en los Estudios Culturales. Revista
Fractal, México, v. 2, nº 6, jul./set. 1997, p. 45-60. Disponível em: <http://www.mxfractal.org/F6cancli. html>. Acesso em: 11 set. 2014. 9 Considerações Finais Os Estudos Culturais, com sua flexibilidade e
abertura a outros campos do conhecimento,
possibilitam leituras dos processos hibridizadores
que se complementam e que também se
confrontam em alguns casos. Entretanto, as
múltiplas leituras da hibridização a partir dos
teóricos da cultura possuem algo em comum
que traduz bem a essência de suas ideias: a
hibridização é ambígua e sempre contraditória. E é justamente por seu caráter dúbio, em especial
pela recusa em pensar apenas nos elementos que
possibilitam a hibridização e que se esquecem dos
resultados, que la différance em Jacques Derrida
é lida e transposta nas concepções dos autores
discutidos neste artigo. Em outros termos, pensar
na dubiedade da ausência e presença, além de
se questionar acerca da existência do diferente
na sociedade, faz com que a hibridização seja
tensionada e sempre refletida na vida dos sujeitos
e nas suas formas de enxergar o mundo. Revista da Associação Nacional dos Programas de Pós-Graduação em Comunicação | E-compós, Brasília, v.18, n.1, jan./abr. 2015. Em outra comparação conceitual, agora com
Homi Bhabha, é possível perceber que também
existem preocupações em torno da linguagem
e vinculadas à hibridização por parte de García
Canclini. A linguagem é apontada pelo argentino
como perturbadora da sociedade normativa, como
intransigente quando os assuntos são regras e
convenções. Ele comenta que as lutas semânticas
para neutralizar, perturbar a mensagem dos
outros ou mudar seu significado, e subordinar
os demais à própria lógica, são elementos de
uma pseudorrealidade, de uma encenação de
conflitos entre os atores políticos e sociais,
isto é, entre a história, o Estado, a publicidade,
o mercado e também a luta popular para
sobreviver. Percebe-se, ainda, que, para García
Canclini, mais importante que configurar o termo
na perspectiva da mestiçagem, crioulização,
sincretismo, é construir princípios teóricos e
procedimentos metodológicos que nos ajudem a
entender as diferenças no sentido de perceber o
que se ganha ou se perde ao hibridar-se. Assim, a visão de Stuart Hall com relação à
hibridização cultural aproxima-se muito da
conceituação proposta por Homi Bhabha,
especialmente por usar uma terminologia
idêntica: a tradução cultural. A seu modo, Hall
define tal processo tradutório como o momento
de negociação entre novas e antigas matrizes
culturais, momentos que são vivenciados por
pessoas que, como ele, emigraram de sua terra
natal. Todavia, os aspectos relativos à linguagem e GARCÍA CANCLINI, N. Consumidores e Cidadãos. Conflitos multiculturais da globalização. 4ª ed. Rio de
Janeiro, Editora UFRJ, 1999. Conflitos multiculturais da globalização. 4ª ed. Rio de
Janeiro, Editora UFRJ, 1999. ______. Culturas híbridas: estratégias para entrar
e sair da modernidade. (Trad. Heloísa P. Cintrão e Ana
Regina Lessa). 4ª ed. São Paulo: Edusp, 2011. ______. Culturas híbridas: estratégias para entrar
e sair da modernidade. (Trad. Heloísa P. Cintrão e Ana
Regina Lessa). 4ª ed. São Paulo: Edusp, 2011. ESCOSTEGUY, A. C. Cartografia dos Estudos
Culturais: uma versão latino-americana. Belo
Horizonte: Autêntica, 2001. GARCÍA CANCLINI, N. Consumidores e Cidadãos. Conflitos multiculturais da globalização. 4ª ed. Rio de
Janeiro, Editora UFRJ, 1999. Referências ______. Noticias recientes sobre hibridación. Biblioteca de Documentos Globalizacion. Org. Texto
apresentado no VI Congreso de la SibE, julho de 2000. Disponível em: http://www.globalizacion.org/biblioteca/
CanciliniHibridacionNoticiasRecientes.htm. Acesso em:
22 de outubro de 2014. ______. Noticias recientes sobre hibridación. Biblioteca de Documentos Globalizacion. Org. Texto
apresentado no VI Congreso de la SibE, julho de 2000. Disponível em: http://www.globalizacion.org/biblioteca/
CanciliniHibridacionNoticiasRecientes.htm. Acesso em:
22 de outubro de 2014. BARBOSA, L. Hall leitor de Derrida. Revista
Urutágua, Maringá, nº 26, maio 2012, p.114-124. Disponível em: <http://periodicos.uem.br/ojs/index. php/Urutagua/article/viewFile/14656/9192>. Acesso
em: 14 de janeiro de 2014. Dessa maneira, como pressuposto básico
das reflexões que seguem o viés dos Estudos
Culturais, seria uma aporia, para fazer uso
de termos derrideanos, dar como encerrada
a discussão conceitual sobre a hibridização
cultural, uma vez que verdades incontestáveis
não fazem parte do escopo teórico dessa rede de
estudos. Pelo contrário, o que se intentou neste
trabalho foi mostrar um panorama, obviamente
incompleto, das variadas concepções que os
processos hibridizadores possuem na atualidade. Uma hibridização que não se limita às fronteiras
geográficas, linguísticas ou de outra ordem, mas
que é fluida e desestabilizadora. ESCOSTEGUY, A. C. Cartografia dos Estudos
Culturais: uma versão latino-americana. Belo
Horizonte: Autêntica, 2001. ESCOSTEGUY, A. C. Cartografia dos Estudos
Culturais: uma versão latino-americana. Belo
Horizonte: Autêntica, 2001. HALL, S. A identidade cultural na pós-modernidade.
3ª ed. Rio de Janeiro: DP&A, 2000. BENNINGTON, G.; DERRIDA, J. Jacques Derrida. (Trad. Anamaria Skinner). Rio de Janeiro: Jorge
Zahar, 1996. BHABA, H. Representation and the Colonial Text: a
critical exploration of some forms of mimeticism. In:
GLOVERSMITH, F. (ed.), The theory of reading. Brigthon: Harvester, 1984, p. 93-122. ______. The Third Space. In: RUTHERFORD, J. (ed.),
Identity: community, culture, difference, London:
Lawrence & Wishart Rutherford, 1990, p 207-215. ______. O local da cultura. Belo Horizonte:
Autêntica, 2010. DERRIDA, J. Margens da filosofia. (Trad. Joaquim T. Costa; Antonio M. Magalhães). Campinas: Papirus, 1991. BENNINGTON, G.; DERRIDA, J. Jacques Derrida. (Trad. Anamaria Skinner). Rio de Janeiro: Jorge
Zahar, 1996. BENNINGTON, G.; DERRIDA, J. Jacques Derrida. (Trad. Anamaria Skinner). Rio de Janeiro: Jorge
Zahar, 1996. Revista da Associação Nacional dos Programas de Pós-Graduação em Com
______. Da diáspora: identidades e mediações
culturais. São Paulo: Humanitas, 2003. JOHNSON, R. O que é, afinal, Estudos Culturais? In: SILVA, T. (Trad./Org.). O que é, afinal, Estudos
Culturais? Belo Horizonte: Autêntica, 2004, p. 9-131. KUNSCH, W. L. Introdução: as matrizes marxistas e
cristãs das idéias comunicacionais na América Latina. In: MELO, J. M.; GOBBI, M. C.; KUNSCH, W. L. (Orgs.). Matrizes comunicacionais Latino-americanas
– Marxismo e Cristianismo. São Paulo: Ed. UMESP/
UNESCO, 2002, p. 13-25. MARTÍN-BARBERO, J. Dos meios às mediações:
comunicação, cultura e hegemonia. (Trad. Ronald BHABA, H. Representation and the Colonial Text: a
critical exploration of some forms of mimeticism. In:
GLOVERSMITH, F. (ed.), The theory of reading. Brigthon: Harvester, 1984, p. 93-122. BHABA, H. Representation and the Colonial Text: a
critical exploration of some forms of mimeticism. In:
GLOVERSMITH, F. (ed.), The theory of reading. Brigthon: Harvester, 1984, p. 93-122. ______. The Third Space. In: RUTHERFORD, J. (ed.),
Identity: community, culture, difference, London:
Lawrence & Wishart Rutherford, 1990, p 207-215. ______. O local da cultura. Belo Horizonte:
Autêntica, 2010. DERRIDA, J. Margens da filosofia. (Trad. Joaquim T. Costa; Antonio M. Magalhães). Campinas: Papirus, 1991. DERRIDA, J. Margens da filosofia. (Trad. Joaquim T. Costa; Antonio M. Magalhães). Campinas: Papirus, 1991. Polito; Sérgio Alcides). 6ª ed. Rio de Janeiro: Editora
UFRJ, 2009. ______. Oficio de cartógrafo: travesías
latinoamericanas de la comunicación en la cultura. Santiago, Chile: Fondo de Cultura Económica, 2002. McCARTHY, C. The uses of culture: education and the
limits of ethnic affiliation. New York: Routledge, 1998. OROZCO GÓMEZ, G. Televisión y producción
de significados. Tres ensayos. Universidad de
Guadalajara, Guadalajara, 1994. PINHEIRO, A. HALL, S. A identidade cultural na pós-modernidade.
3ª ed. Rio de Janeiro: DP&A, 2000. Aquém da Identidade e da Oposição:
Formas na cultura mestiça. 2ª ed. Piracicaba: Editora
UNIMEP, 1995. ______. Oficio de cartógrafo: travesías
latinoamericanas de la comunicación en la cultura. Santiago, Chile: Fondo de Cultura Económica, 2002. McCARTHY, C. The uses of culture: education and the
limits of ethnic affiliation. New York: Routledge, 1998. OROZCO GÓMEZ, G. Televisión y producción
de significados. Tres ensayos. Universidad de
Guadalajara, Guadalajara, 1994. Revista da Associação Nacional dos Programas de Pós-Graduação em Comunicação | E-compós, Brasília, v.18, n.1, jan./abr. 2015. 16/18 PINHEIRO, A. Aquém da Identidade e da Oposição:
Formas na cultura mestiça. 2ª ed. Piracicaba: Editora
UNIMEP, 1995. SANTIAGO, S. Glossário de Derrida. Rio: Francisco
Alves, 1976. SOUZA, L. M. T. M. Hibridismo e tradução cultural em
Bhabha. In: ABDALA JÚNIOR, B. (Org.). Margens da
cultura: mestiçagem, hibridismo & outras misturas. São Paulo: Boitempo Editorial, 2004, p. 113-132. SOUZA, L. M. T. M. Hibridismo e tradução cultural em TORRES, S. Crónicas de viaje chicanas: The
Mixquiahuala Letters de Ana Castillo y Paletitas de
guayaba de Erlinda González-Berry. In: DRISCOLL et
al. Límites sociopoliticos y fronteras culturales en
América del Norte. Cidade do México: UNAM/CISAN,
2000, p. 161-177. www.e-compos.org.br
| E-ISSN 1808-2599 | Comunicando diferencias:
los procesos de hibridación
bajo la comprensión de la différance
en los Estudios Culturales Abstract The article discusses the process of hybridization
and the multiple readings of what being or
becoming hybrid is. The approach starts with
Jacques Derrida, with the discussion of the divided
nature of the sign, “la différance”, and continues
by the consensual understandings (and some
dissents) to theorists such as Stuart Hall, Homi
Bhabha, Martín-Barbero and García Canclini –
locating, whenever possible, the reflections on
the context of Latin America. We conclude that
hybridization is potentially creative and at the same
time troubled, not being limited to geographical,
linguistic or any other border, but fluid, unsettling,
ambiguous and almost often controversial. El artículo analiza el proceso de hibridación y
múltiples lecturas de lo que el ser o llegar a ser
híbrido. El enfoque comienza por Jacques Derrida,
con la discusión sobre la naturaleza dividida de la
señal, la différance, y continúa por entendimientos
consensuales (y un poco de la disidencia) a teóricos
como Stuart Hall, Homi Bhabha, Martín-Barbero
y García Canclini - siempre que sea posible con
reflexiones en el contexto de América Latina. Llegamos a la conclusión de que la hibridación
es potencialmente creativo y al mismo tiempo
preocupado, no se limita a las fronteras geográficas,
lingüísticas o de otro tipo, y es fluida, inquietante,
ambigua y, a menudo contradictorias. Revista da Associação Nacional dos Programas de Pós-Graduação em Comunicação | E-compós, Brasília, v.18, n.1, jan./abr. 2015. 17/18 Communicating differences:
hybridization processes from
the la différance conception
in Cultural Studies Comunicando diferencias:
los procesos de hibridación
bajo la comprensión de la différance
en los Estudios Culturales E-COMPÓS | www.e-compos.org.br | E-ISSN 1808-2599 E-COMPÓS | www.e-compos.org.br | E-ISSN 1808-2599 Expediente Revista da Associação Nacional dos Programas
de Pós-Graduação em Comunicação. Brasília, v.18, n.1, jan./abri.. 2015. A identificação das edições, a partir de 2008,
passa a ser volume anual com três números. A revista E-Compós é a publicação científica em formato eletrônico
da Associação Nacional dos Programas de Pós-Graduação em
Comunicação (Compós). Lançada em 2004, tem como principal
finalidade difundir a produção acadêmica de pesquisadores da área
de Comunicação, inseridos em instituições do Brasil e do exterior. Itania Maria Mota Gomes, Universidade Federal da Bahia, Brasil
Janice Caiafa, Universidade Federal do Rio de Janeiro, Brasil
Jiani Adriana Bonin, Universidade do Vale do Rio dos Sinos, Brasil
José Afonso da Silva Junior, Universidade Federal de Pernambuco, Brasil
José Luiz Aidar Prado, Pontifícia Universidade Católica de São Paulo, Brasil
Kati Caetano, Universidade Tuiuti do Paraná, Brasil
Lilian Cristina Monteiro França, Universidade Federal de Sergipe, Brasil
Liziane Soares Guazina, Universidade de Brasília, Brasil
Luíza Mônica Assis da Silva, Universidade de Caxias do Sul, Brasil
Luciana Miranda Costa, Universidade Federal do Pará, Brasil
Malena Segura Contrera, Universidade Paulista, Brasil
Marcel Vieira Barreto Silva, Universidade Federal da Paraíba, Brasil
Maria Ogécia Drigo, Universidade de Sorocaba, Brasil
Maria Ataide Malcher, Universidade Federal do Pará, Brasil
Maria Clotilde Perez Rodrigues, Universidade de São Paulo, Brasil
Maria das Graças Pinto Coelho, Universidade Federal do Rio Grande
do Norte, Brasil Keywords Hibridación. Estudios Culturales. Revista da Associação Nacional dos Programas de Pós-Graduação em Comunicação | E-compós, Br
Recebido em:
02 de março de 2014
Aceito em:
27 de fevereiro de 2015
Communication of differences. La différance. Comunicación de la diferencia. La différance. Recebido em:
02 de março de 2014
Aceito em:
27 de fevereiro de 2015 CONSELHO EDITORIAL Alexandre Rocha da Silva, Universidade Federal do Rio Grande do Sul, Brasil
Alexandre Farbiarz, Universidade Federal Fluminense, Brasil
Ana Carolina Damboriarena Escosteguy, Pontifícia Universidade
Católica do Rio Grande do Sul, Brasil
Ana Carolina Rocha Pessôa Temer, Universidade Federal de Goiás, Brasil
Ana Regina Barros Rego Leal, Universidade Federal do Piauí, Brasil
André Luiz Martins Lemos, Universidade Federal da Bahia, Brasil
Andrea França, Pontifícia Universidade Católica do Rio de Janeiro, Brasil
Antonio Carlos Hohlfeldt, Pontifícia Universidade Católica do Rio Grande
do Sul, Brasil Ana Carolina Rocha Pessôa Temer, Universidade Federal de Goiás, Brasil
Ana Regina Barros Rego Leal, Universidade Federal do Piauí, Brasil
André Luiz Martins Lemos, Universidade Federal da Bahia, Brasil
Andrea França, Pontifícia Universidade Católica do Rio de Janeiro, Brasil
Antonio Carlos Hohlfeldt, Pontifícia Universidade Católica do Rio Grande
do Sul, Brasil Arthur Ituassu, Pontifícia Universidade Católica do Rio de Janeiro, Brasil
Álvaro Larangeira, Universidade Tuiuti do Paraná, Brasil
Ângela Freire Prysthon, Universidade Federal de Pernambuco, Brasil
César Geraldo Guimarães, Universidade Federal de Minas Gerais, Brasil
Cláudio Novaes Pinto Coelho, Faculdade Cásper Líbero, Brasil
Daisi Irmgard Vogel, Universidade Federal de Santa Catarina, Brasil
Daniela Zanetti, Universidade Federal do Espírito Santo, Brasil
Denize Correa Araujo, Universidade Tuiuti do Paraná, Brasil
Eduardo Antonio de Jesus, Pontifícia Universidade Católica
de Minas Gerais, Brasil Mauricio Ribeiro da Silva, Universidade Paulista, Brasil
Mauro de Souza Ventura, Universidade Estadual Paulista, Brasil
Márcio Souza Gonçalves, Universidade do Estado do Rio de Janeiro, Brasil
Micael Maiolino Herschmann, Universidade Federal do Rio de Janeiro, Brasil
Mirna Feitoza Pereira, Universidade Federal do Amazonas, Brasil
Nísia Martins Rosario, Universidade Federal do Rio Grande do Sul, Brasil
Potiguara Mendes Silveira Jr, Universidade Federal de Juiz de Fora, Brasil
Regiane Ribeiro, Universidade Federal do Paraná, Brasil
Rogério Ferraraz, Universidade Anhembi Morumbi, Brasil
Rose Melo Rocha, Escola Superior de Propaganda e Marketing, Brasil
Rozinaldo Antonio Miani, Universidade Estadual de Londrina, Brasil
Sérgio Luiz Gadini, Universidade Estadual de Ponta Grossa, Brasil
Simone Maria Andrade Pereira de Sá, Universidade Federal Fluminense, Brasil
Veneza Mayora Ronsini, Universidade Federal de Santa Maria, Brasil
Walmir Albuquerque Barbosa, Universidade Federal do Amazonas, Brasil Eduardo Vicente, Universidade de São Paulo, Brasil
Elizabeth Moraes Gonçalves, Universidade Metodista de São Paulo, Brasil
Erick Felinto de Oliveira, Universidade do Estado do Rio de Janeiro, Brasil
Francisco Elinaldo Teixeira, Universidade Estadual de Campinas, Brasil
Francisco Paulo Jamil Almeida Marques, Universidade Federal do Ceará, Brasil
Gabriela Reinaldo, Universidade Federal do Ceará, Brasil
Gisela Grangeiro da Silva Castro, Escola Superior de Propaganda
e Marketing, Brasil
Goiamérico Felício Carneiro Santos, Universidade Federal de Goiás, Brasil
Gustavo Daudt Fischer, Unisinos, Brasil
Herom Vargas, Universidade Municipal de São Caetano do Sul, Brasil Revista da Associação Nacional dos Programas de Pós-Gra
COMPÓS | www.compos.org.br
Associação Nacional dos Programas de Pós-Graduação em
Comunicação
Presidente
Eduardo Morettin
Universidade de São Paulo, Brasil
eduardomorettin@usp.br
Vice-presidente
Inês Vitorino
Universidade Federal do Ceará, Brasil
ines@ufc.br
Secretária-Geral
Gislene da Silva
Universidade Federal de Santa Catarina, Brasil
gislenedasilva@gmail.com Revista da Associação Nacional dos Programas de Pós-Gra
COMPÓS | www.compos.org.br
Associação Nacional dos Programas de Pós-Graduação em
Comunicação
Presidente
Eduardo Morettin
Universidade de São Paulo, Brasil
eduardomorettin@usp.br
Vice-presidente
Inês Vitorino
Universidade Federal do Ceará, Brasil
ines@ufc.br
Secretária-Geral
Gislene da Silva
Universidade Federal de Santa Catarina, Brasil
gislenedasilva@gmail.com www.e-compos.org.br
| E-ISSN 1808-2599 | E-COMPÓS | www.e-compos.org.br | E-ISSN 1808-2599 COMISSÃO EDITORIAL COMISSÃO EDITORIAL
Cristiane Freitas Gutfreind
Pontifícia Universidade Católica do Rio Grande do Sul, Brasil
Irene Machado
Universidade de São Paulo, Brasil
Jorge Cardoso Filho
Universidade Federal do Reconcavo da Bahia, Brasil
Universidade Federal da Bahia, Brasil
Revisão de textos | Press Revisão
SECRETÁRIA EXECUTIVA | Helena Stigger
EDITORAÇÃO ELETRÔNICA | Roka Estúdio Cristiane Freitas Gutfreind
Pontifícia Universidade Católica do Rio Grande do Sul, Brasil
Irene Machado
Universidade de São Paulo, Brasil
Jorge Cardoso Filho
Universidade Federal do Reconcavo da Bahia, Brasil
Universidade Federal da Bahia, Brasil
Revisão de textos | Press Revisão
SECRETÁRIA EXECUTIVA | Helena Stigger
EDITORAÇÃO ELETRÔNICA | Roka Estúdio Revisão de textos | Press Revisão
SECRETÁRIA EXECUTIVA | Helena Stigger
EDITORAÇÃO ELETRÔNICA | Roka Estúdio Revisão de textos | Press Revisão
SECRETÁRIA EXECUTIVA | Helena Stigger
EDITORAÇÃO ELETRÔNICA | Roka Estúdio
|
https://openalex.org/W2945016456
|
https://europepmc.org/articles/pmc6896980?pdf=render
|
English
| null |
Examining differences in out-of-hours primary care use in Belgium and the Netherlands: a cross-sectional study
|
European journal of public health
| 2,019
|
cc-by
| 7,442
|
Marleen Smits1,2,, Annelies Colliers3,, Tessa Jansen1, Roy Remmen3,
Stephaan Bartholomeeusen3, Robert Verheij1 1 Netherlands Institute for Health Services Research (Nivel), Utrecht, The Netherlands 1 Netherlands Institute for Health Services Research (Nivel), Utrecht, The Netherlands
2 Scientific Center for Quality of Healthcare (IQ Healthcare), Radboud Institute for Health Sciences, Radboud University
Medical Center, Nijmegen, The Netherlands
3 Department of Primary and Interdisciplinary Care (ELIZA), Centre for General Practice, Faculty of Medicine and Health
Sciences, University of Antwerp, Antwerp, Belgium 1 Netherlands Institute for Health Services Research (Nivel), Utrecht, The Netherlands
2 Scientific Center for Quality of Healthcare (IQ Healthcare), Radboud Institute for Health Sciences, Radboud University
Medical Center, Nijmegen, The Netherlands
3 Department of Primary and Interdisciplinary Care (ELIZA), Centre for General Practice, Faculty of Medicine and Health
Sciences, University of Antwerp, Antwerp, Belgium 1 Netherlands Institute for Health Services Research (Nivel), Utrecht, The Netherlands
2 Scientific Center for Quality of Healthcare (IQ Healthcare), Radboud Institute for Health Sciences, Radboud University
Medical Center, Nijmegen, The Netherlands
3 Department of Primary and Interdisciplinary Care (ELIZA), Centre for General Practice, Faculty of Medicine and Health
Sciences, University of Antwerp, Antwerp, Belgium 1 Netherlands Institute for Health Services Research (Nivel), Utrecht, The Netherlands
2 Scientific Center for Quality of Healthcare (IQ Healthcare), Radboud Institute for Health Sciences, Radboud University
Medical Center, Nijmegen, The Netherlands
3 Department of Primary and Interdisciplinary Care (ELIZA) Centre for General Practice Faculty of Medicine and Health ,
j
g
,
3 Department of Primary and Interdisciplinary Care (ELIZA), Centre for General Practice, Faculty of Medicine
Sciences, University of Antwerp, Antwerp, Belgium These authors contributed equally to this article. Correspondence: Marleen Smits, Radboud University Medical Center, P.O. Box 9101, 114 IQ Healthcare, 6500 HB
Nijmegen, The Netherlands, Tel: +31 (0) 24 36 66264, e-mail: marleen.smits@radboudumc.nl Background: The organizational model of out-of-hours primary care is likely to affect healthcare use. We aimed to
examine differences in the use of general practitioner cooperatives for out-of-hours care in the Netherlands and
Belgium (Flanders) and explore if these are related to organizational differences. Methods: A cross-sectional
observational study using routine electronic health record data of the year 2016 from 77 general practitioner
cooperatives in the Netherlands and 5 general practitioner cooperatives in Belgium (Flanders). Patient age, gender
and health problem were analyzed using descriptive statistics. Results: The number of consultations per 1000
residents was 2.3 times higher in the Netherlands than in Belgium. 1018 Importance for the elderly of discussion in the
family and society about attitude toward organ donation. Transplant Proc 2018;50:523–5. 40 Febrero B, Rı´os A, Martı´nez-Alarco´n L, et al. Information received by secondary
school teaching personnel about organ donation and transplantation: a study in the
southeast of Spain. Transplant Proc 2013;45:3589–91. 31 Gross T, Martinoli S, Spagnoli G, et al. Attitudes and behavior of young European
adults towards the donation of organs—a call for better information. Am J
Transplant 2001;1:74–81. The Author(s) 2019. Published by Oxford University Press on behalf of the European Public Health Association. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/
4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. doi:10.1093/eurpub/ckz083
Advance Access published on 13 May 2019 1018 1018 European Journal of Public Health 32 Rı´os A, Febrero B, Lo´pez-Navas A, et al. From where do our children receive infor-
mation about organ donation and transplantation? Transplant Proc 2010;42:3113–5. 24 Rı´os A, Febrero B, Lo´pez-Navas A, et al. Xenotransplantation of cells for diabetes: Is
there a good acceptance rate among Spanish adolescents? Transplant Proc
2011;43:55–7. 33 Febrero B, Almela J, Rı´os A, et al. Teenagers in rural areas faced with organ donation
and transplantation. Transplant Proc 2018;50:520–2. 25 Almela-Baeza J, Febrero-Sa´nchez B, Pe´rez-Manzano A, Ramı´rez-Romero P. Tema´ticas prosociales e impacto de una intervencio´n educativa Flipped Classroom
relacionada con la promocio´n de la salud mediante la creacio´n relatos audiovisuales. Rev Mediterra´nea Comun 2018;9:75–84. 34 Cardenas V, Thornton JD, Wong KA, et al. Effects of classroom education on
knowledge and attitudes regarding organ donation in ethnically diverse urban high
schools. Clin Transpl 2010;24:784–93. 26 Rı´os A, Febrero B, Lo´pez-Navas A, et al. Evaluation of attitude toward living organ
donation among adolescents in Southeast Spain. Transplant Proc 2012;44:1489–92. 35 Febrero B, Rı´os A, Martı´nez-Alarco´n L, et al. Knowledge of the brain death concept
among adolescents in southeast Spain. Transplant Proc 2013;45:3586–8. 27 Rı´os A, Febrero B, Lo´pez-Navas A, et al. Secondary school teachers’ assessment of
the introduction of an educational program about organ donation and trans-
plantation. Transplant Proc 2011;43:58–60. 36 Rı´os A, Febrero B, Lo´pez-Navas A, et al. Knowledge of the brain-death concept
among secondary school teachers. Transplant Proc 2012;45:1486–8. 37 Rı´os A, Martı´nez-Alarco´n L, Lo´pez-Navas A, et al. Knowledge of the concept of
brain death in the British and Irish population resident in southeast Spain. Transplant Proc 2011; 43:692–4. 28 Piccoli GB, Soragna G, Putaggio S, et al. Efficacy of an educational programme for
secondary school students on opinions on renal transplantation and organ
donation: a randomized controlled trial. Nephrol Dial Transplant 2006;21:499–509. 38 Waldrop DP, Tamburlin JA, Thompson SJ, Simon M. Life and death decisions:
using school-based health education to facilitate family discussion about organ and
tissue donation. Death Stud 2004;28:643–57. 29 Rey JW, Grass V, Barreiros AP, et al. Organ procurement in Germany: a regional
survey among students. Dtsch Med Wochenschr 2012;137:69–73. 30 Weaver M, Spigner C, Pineda M, et al. Knowledge and opinions about organ
donation among urban high school students: pilot test of a health education
program. Clin Transpl 2000;14:292–303. 39 Febrero B, Almela J, Febrero R, et al. Examining differences in out-of-hours primary care use
in Belgium and the Netherlands: a cross-sectional study Marleen Smits1,2,, Annelies Colliers3,, Tessa Jansen1, Roy Remmen3,
Stephaan Bartholomeeusen3, Robert Verheij1 Introduction for completeness of the data, i.e. having registration data from
52 weeks, with at least 500 contacts per week. To determine the
number of residents in the catchment area and the gender and age
distributions of the population, we used data from Statistics
Netherlands (CBS) at the level of four-digit postal code areas
belonging to the adherence area of the GPC. I
n many western countries, organizing 24/7 care for patients is an
important task for primary care, that can be performed in many
different ways.1,2 Out-of-hours (OOH) primary care is continuously
evolving, driven by issues such as growing patient demands, an aging
population, an acceptable work-life balance for general practitioners
(GPs) and financial strains. Therefore, the organization of OOH
primary care is a focus for many healthcare professionals, policy
makers and researchers throughout Europe.3 In Belgium, the data were provided by iCAREdata-database
(Improving Care And Research Electronic Data Trust Antwerp), a
research database linking and collecting routine data from GPCs,
pharmacies and Emergency Departments (EDs).9 In 2016, 7 out of
the 26 Flemish GPCs delivered data to iCAREdata. For this purpose,
five GPCs were included, selected on the basis of completeness of
data. Some of the characteristics of OOH care may have an impact on
patients’ or healthcare professionals’ satisfaction, workload, patient
safety, quality of care and healthcare costs. In addition, it influences
patients’ threshold to seek care and, consequently, the use of OOH
care.4 We extracted patient age (years), gender (male/female), health
problem (ICPC code, International Classification of Primary
Care), date and time and contact type (telephone contact/face-to-
face clinic consultation/home visit). To improve the comparability
of both datasets, we excluded the following contacts: (i) home visits,
because home visits are not always recorded in Belgium, (ii) contacts
with patients aged >75 years, because in Belgium, these patients are
almost always seen during home visits and (iii) Dutch GPC contacts
outside the opening hours of the Belgian GPC: for optimal
comparison we only used data from the weekends (19 p.m. on
Fridays until 7 a.m. on Mondays) in both countries. For the
figures on health problems, there was a second selection criterion
for GPCs, that differed per country. NIVEL Primary Care Database
uses the criterion that only GPCs are included at which a meaningful
ICPC code was recorded for at least 70% of the contacts. Design and population We performed a cross-sectional observational study using routinely
recorded electronic health record data from GPCs. Data were
derived from 77 GPCs spread across the Netherlands, covering a
population of 11 300 000 (67% of the Dutch population), and 5
GPCs in Belgium (region of Flanders; i.e. the Northern part of
Belgium) covering a population of 815 000 (11% of the Flemish
population). Introduction The codes
A97 (no disease) and A99 (other generalized disease) are not
regarded as a meaningful.10 For iCAREdata, GPCs that were
included in the analyses of health problems had an ICPC code
recorded for at least 90% of contacts. Although Belgium and the Netherlands are two neighboring
West-European countries, the developments in OOH care have
not evolved in parallel. In the Netherlands, Dutch GPs reorganized
OOH primary care around the year 2000 shifting from small
rotation groups to large-scale GP cooperatives (GPCs) in which
50–250 GPs take care of populations ranging from 100 000–500
000 citizens.5 Since the last two decades, Belgian OOH care is im-
plementing changes in the region of Flanders based on the Dutch
system as a role model.6 Still there are many differences. The most
pronounced differences for GPCs in Belgium compared with Dutch
GPCs are a lack of a gate keeper role for the GP to secondary or
tertiary care, co-payment by patients and the restricted opening
hours (only weekends and public holidays). In addition, in the
Netherlands, patients are expected to contact the GPC first by
phone, and are triaged by trained professionals using a protocolized
triage system. One of the possibilities for follow-up is to give patients
telephone advice. Self-care advice or postponement of care to
daytime care is possible, while in Belgium the GP should see every
patient and these options are not possible. Table 1 shows the main
organizational differences between the OOH services in Belgium
(Flanders) and the Netherlands. Ethical approval
Belgium We aim to examine differences in the use of GPCs for OOH
primary care in Belgium (Flanders) and the Netherlands and
explore if these are related to organizational differences. The infor-
mation can be useful for healthcare professionals and policy makers
engaged in designing the most optimal organizational model for
OOH primary care. Ethical approval for data extraction from the electronic medical
records in the iCAREdata database was granted by the Ethics
Committee of the University of Antwerp/University Hospital
Antwerp (12/49/404 and 13/34/330). To secure the privacy of infor-
mation about individual patients, a permission for the data
collection and data-linking was obtained from the Committee of
Health of the Commission for the Protection of Privacy (No. 14/
094 n173 and No. 14/194 n133). The Netherlands The study was approved according to the governance code of NIVEL
Primary Care Database, Utrecht (NZR-00318.007). GPCs that par-
ticipate are contractually obliged to: (i) inform their patients about
their participation in NIVEL Primary Care Database and (ii) to
inform patients about the option to opt-out for inclusion of their
data in the database. Dutch law allows the use of electronic health
records for research purposes under certain conditions. Neither
obtaining informed consent from patients nor approval by a
medical ethics committee is obligatory for observational studies
containing no directly identifiable data (Dutch Civil Law, Article
7: 458). The residents in the catchment area of the Dutch GPCs were
representative for the general Dutch population in terms of age
and gender. However, there was a slight over-representation of
residents in strongly urbanized areas.8 In Belgium, both rural and
urban GPCs were represented, and residents were representative for
the Flemish population in terms of age and gender. In both countries, data were only included from patients who did
not object to the use of their data. To guarantee the privacy of the
patients, the researchers had no access to direct identifiable patient
information, such as name, address or citizen service number. Identifiable patient information was converted to a so-called
pseudonym by software of a Trusted Third Party: a non-profit
foundation called ZorgTTP in the Netherlands and a public govern-
mental institution called e-Health in Belgium. Marleen Smits1,2,, Annelies Colliers3,, Tessa Jansen1, Roy Remmen3,
Stephaan Bartholomeeusen3, Robert Verheij1 Excluding telephone consultations, which are
not possible in Belgium, the number of consultations was 1.4 times higher. In Belgium, the top 10 of health
problems was mainly related to infections, while in the Netherlands there were a larger variety of health
problems. In addition, the health problem codes in the Dutch top 10 were more often symptoms, while the
codes in the Belgian top 10 were more often diagnoses. In both countries, a relatively large percentage of GPC
patients were young children and female patients. Conclusion: Differences in the use of general practitioner
cooperatives seem to be related to the gatekeeping role of general practitioners in the Netherlands and to
organizational differences such as telephone triage, medical advice by telephone, financial thresholds and
number of years of experience with the system. The information can benefit policy decisions about the organiza-
tion of out-of-hours primary care. 1019 Examining differences in out-of-hours primary care use in Belgium and the Netherlands The Netherlands
The GP is the point of access to secondary care but
patients in need for highly acute care can go to the
ED without prior contact with the GP or GPC. Also
access via ambulance.
Professionals: emergency physicians, resident physicians
and nurses.
There is a trend of co-location and collaboration
between the ED and the GPC during OOH.
Access with central phone number 112.
Triage by nurses using a triage system (in 40% this is the
same system as the GPC), ambulance manned by nurse
and driver who can give medical assistance.
Nurse on ambulance decides to treat the patient or
transport to hospital.
General practices are closed during OOH.
78% of the patients have contact with their GP at least
once a year.
Average annual consultation rate per individual is 4.4.8
Nearly all hospitals have an ED.
Access with central phone number 112.
Triage by trained non-medical staff, using a
triage system, paramedics, upscaling possible
with medical doctor and ED nurse when
necessary
All patient are transported to the hospital.
General practices are closed during OOH.
79% of the patients have contact with their GP
at least once a year.
Average annual consultation rate per individual
is 4.3.7
Access with central phone number 112.
Triage by trained non-medical staff, using a
triage system, paramedics, upscaling possible
with medical doctor and ED nurse when
necessary
All patient are transported to the hospital.
General practices are closed during OOH.
79% of the patients have contact with their GP
at least once a year.
Average annual consultation rate per individual
is 4.3.7 Daytime general practices
General practice
General practices are closed during OOH.
78% of the patients have contact with their GP at least
once a year. y
Average annual consultation rate per individual is 4.4.8 Data analysis patient can have more than one contact per year. Differences
between the two countries were not tested for statistical significance
because of the large sample size. Figures were produced using R
version 3.4.2. We performed descriptive analyses using statistical software Stata,
version 14 for the Dutch data and Microsoft SQL server 2012 for the
Belgian data. We analyzed patient age, gender and health problems
(ICPC). For the comparison of the age distribution of GPC patients
with the age distribution of the population, we used patients as unit
of analysis. For all other analyses, we used contacts as units of
analysis to gain insight in the total amount of healthcare use; a The Netherlands Type of healthcare professionals
available
GPs, assisted by chauffeurs for home visits
Telephone triage: assessment of
urgency and appropriate
type of care
Patients can walk in or call the GPC in advance. Telephone call always results in face-to-face con-
sultation. No telephone triage. Pilot project in one region with telephone triage by
112 operator, with one central number (1733). Payment GP
Varies between GPCs: mostly fee-for-service (37.5E
per daytime consultation, 56E per daytime home
visit; higher rates in the night), salary per hour or
capitation based. Patient co-payments
Direct payment, with partly reimbursement by
obligatory health insurance or third party
payment (mutuality). Number of inhabitants in
catchment area per GPC
80 000–150 000
Opening hours OOH services
From Friday 7 p.m. until Monday 7 a.m. and on
public holidays. Related out-of-hours acute
healthcare services
Emergency care
Nearly all hospitals have an ED.
Free access, no referral necessary, secondary and
tertiary care facilities. Also access via ambulance.
Professionals: emergency physicians, resident
physicians and nurses. Ambulance care
Access with central phone number 112.
Triage by trained non-medical staff, using a
triage system, paramedics, upscaling possible
with medical doctor and ED nurse when
necessary
All patient are transported to the hospital. Daytime general practices
General practice
General practices are closed during OOH.
79% of the patients have contact with their GP
at least once a year.
Average annual consultation rate per individual
is 4.3.7 GPs, assisted by chauffeurs for home visits (
)
GPs, chauffeurs, triagists (assistants/nurses), sometimes
nurse practitioners or physician assistants who do con-
sultations for some types of health problems. Access via a regional telephone number (only 5–10% walk
in without a call in advance). Telephone triage by triagists, supervised by a GP. Salary per hour (69E per hour for GP practice owners;
variable rates for locum GPs). Free: obligatory national health insurance scheme includes
general practice care. Deductible excess is not applicable
for GP service (OOH as well as daytime care). 100 000–500 000 Free: obligatory national health insurance scheme includes
general practice care. Deductible excess is not applicable
for GP service (OOH as well as daytime care). 100 000–500 000 Daily from 5 p.m. to 8 a.m. and the entire weekend and on
public holidays. From Friday 7 p.m. until Monday 7 a.m. and on
public holidays.
Nearly all hospitals have an ED. Procedure We used data from electronic health records from 1 January to 31
December 2016. In the Netherlands, we used data provided by
NIVEL Primary Care Database. This database includes routinely
recorded electronic health records from general practices, allied
healthcare practices and 85 of the 120 GPCs in the Netherlands.8
Of these 85 GPCs, 77 GPCs were included that satisfied the criteria European Journal of Public Health 1020 Table 1 Characteristics of out-of-hours care in Belgium (Flanders) and the Netherlands Table 1 Characteristics of out-of-hours care in Belgium (Flanders) and the Netherla
Belgium (Flanders)
Out-of-hours primary care
Organizational model(s)
Small rotation groups or large-scale organizations
(50%): general practitioner cooperatives. Description of model
Unrestricted access to any primary, secondary
and tertiary care facility.
26 GPCs in Flanders, 50% of residents are
covered, 80–160 GPs per GPC.
All GPs are obliged to participate in the on call
system irrespective of their practice size.
GPC location usually not near ED.
Consultation in GPC (80%) or home visit (20%). Telephone consultations not provided. The Netherlands Daytime general practices
General practice The Netherlands Large-scale organizations: general practitioner coopera-
tives. A uniform system nation-wide. Large-scale organizations: general practitioner coopera-
tives. A uniform system nation-wide.
GP has a gate keeper role to secondary and tertiary
care. There is a list system: patients are listed as
patients in a particular practice; this is a consequence of
the reimbursment system (combination of capitation
and fee-for-service).
120 GPCs; 50–250 GPs per GPC.
GP practice owners are obliged to do shifts according to
their practice size. All GPs have to do a minimum
number of shifts to maintain registration as a GP. GPs
are 4 h/week on call, and do 85% of shifts themselves
(15% by locum GPs).
65%
of
GPCs
are
co-located
with
Emergency
Department (ED). Type of healthcare professionals
available
GPs, assisted by chauffeurs for home visits
Telephone triage: assessment of
urgency and appropriate
type of care
Patients can walk in or call the GPC in advance. Telephone call always results in face-to-face con-
sultation. No telephone triage. Pilot project in one region with telephone triage by
112 operator, with one central number (1733). Payment GP
Varies between GPCs: mostly fee-for-service (37.5E
per daytime consultation, 56E per daytime home
visit; higher rates in the night), salary per hour or
capitation based. Patient co-payments
Direct payment, with partly reimbursement by
obligatory health insurance or third party
payment (mutuality). Number of inhabitants in
catchment area per GPC
80 000–150 000
Opening hours OOH services
From Friday 7 p.m. until Monday 7 a.m. and on
public holidays. Related out-of-hours acute
healthcare services
Emergency care
Nearly all hospitals have an ED.
Free access, no referral necessary, secondary and
tertiary care facilities. Also access via ambulance.
Professionals: emergency physicians, resident
physicians and nurses. Ambulance care
Access with central phone number 112.
Triage by trained non-medical staff, using a
triage system, paramedics, upscaling possible
with medical doctor and ED nurse when
necessary
All patient are transported to the hospital. Daytime general practices
General practice
General practices are closed during OOH.
79% of the patients have contact with their GP
at least once a year.
Average annual consultation rate per individual
is 4.3.7
Telephone advice (40%), center consultation (50%) or
home visit (10%). Results Moreover, Dutch GPCs are often co-located
with an ED.15 Patients who walk in with the intention to visit the ED
may be referred to the GPC instead. The Belgian GP has no official
gate keeper function and the threshold for patients to visit the ED is
low.16 Having a gatekeeping system could lead to a higher use of
primary care. Finally, the financial barrier to contact the GPC is
lower in the Netherlands than in Belgium. GPC use is financially
covered by the Dutch health insurance system. Belgian patients do
have to pay the GPC and there is only partly reimbursement by the
health insurance. In both countries, a larger percentage of the patients that consult
the GPC are children between the age of 0 and 4 years compared
with the age distribution of the total population. To a lesser extent,
this also holds for the category 25–45 years. In contrast, fewer people
in the age categories from 45 years and above consulted the GPC as
compared with the age distribution of the population (figure 2). Table 2 shows the 10 most frequently used ICPC-codes for face-
to-face clinic consultations at the GPC in Belgium and the
Netherlands. The most frequently used ICPC code for Belgian
face-to-face clinic consultations at the GPC was R74 (acute upper
respiratory infection; 12.7%), while in the Netherlands this code was
used in 4.5% of this type of consultations. In the Netherlands, S18
(Laceration/cut) was the most frequently used ICPC code (6.3%),
compared with 2.5% in Belgium. In Belgium, the first 8 ICPCs in the
top 10 are related to infections, while in the Netherlands the ICPC
codes show a larger variation. In addition, the ICPC codes in the
Dutch top 10 are more often symptoms, while the ICPC codes in the
Belgian top 10 are more often diagnoses. Supplementary table S1 shows the top 10 ICPC codes in telephone
consultations for the Netherlands only, because in Belgium, a
telephone contact would always result in a face-to-face contact
with the GP. The most frequently used ICPC code for Dutch
telephone consultations was A03 (fever; 4.0%). In the Netherlands as well as in Belgium, a relatively large number
of children between the age of 0 and 4 years consult the GPC. Results For the Netherlands, we included 1 440 517 contacts and for
Belgium 46 973 contacts. The mean number of consultations per 1021 Examining differences in out-of-hours primary care use in Belgium and the Netherlands Figure 1 Number of consultations per 1000 residents at the GPC by gender. Belgium: N=46 973 contacts; The Netherlands: N=1 440 517
contacts Figure 1 Number of consultations per 1000 residents at the GPC by gender. Belgium: N=46 973 contacts;
contacts Figure 1 Number of consultations per 1000 residents at the GPC by gender. Belgium: N=46 973 contacts; The Netherlands: N=1 440 517
contacts ons per 1000 residents at the GPC by gender. Belgium: N=46 973 contacts; The Netherlands: N=1 440 517 1000 residents of the GPC regions was 2.3 times higher in the
Netherlands (145 per 1000 residents) than in Belgium (63 per
1000 residents). Excluding telephone consultations, which are not
possible in Belgium, the number of consultations was 1.4 times
higher in the Netherlands (88 per 1000 residents) than in Belgium. In both countries, more female than male patients consulted the
GPC (figure 1). eventually lead to additional face-to-face consultations.11 Belgian law
does not permit medical advice by telephone and all patients are
seen by the GP. Excluding telephone consultations, the number of
consultations was 1.4 times higher in the Netherlands than in
Belgium. Secondly, the system of OOH primary care organized in
GPCs exists much longer in the Netherlands and the GPC is now a
well-known care provider. In the past 15 years, the number of
patients contacting the GPC has increased,5 while ED utilization
decreased.12 In Belgium, when implementing the first GPCs, they
also expected a patient shift from EDs to GPCs. However, patient
numbers at EDs remained stable and increased at GPCs,6,13 probably
because there was still free access to the ED. Thus, the introduction
of a new organization within the healthcare system can lead to more
help seeking of patients and an increased use over time, also called
supplier-induced demand.14 Thirdly, Dutch GPs have a gatekeeping
role with respect to secondary care: to access a medical specialist, a
referral from the GP is needed. Patients can walk in at the ED, but
this is discouraged (e.g. by a national radio campaign) and often
they first contact a GP. Results Previous studies have also reported that relatively many young
children contact OOH primary care,13,17–21 probably as a conse-
quence of the parental needs for reassurance and information on
self-management strategies and mostly related to childhood fever
and common infections.17 In Belgium, there is a long tradition of
home visits, although the numbers are decreasing over time. Largest
part of the home visits are for the elderly. These contacts could be
interesting in the context of the growing group of (frail) elderly.22
Because of the lack of registrations we excluded home visits and
patients >75 years for this study. Main findings and interpretation The use of GPCs for OOH primary care differs between the
Netherlands and Belgium. A prominent difference is that the
number of consultations per 1000 patients is 2.3 times higher in
the Netherlands than in Belgium. The difference in healthcare use
could hypothetically be related to differences in the organization of
the GPCs and national healthcare systems. Firstly, the possibility to
seek medical advice by telephone can lower the threshold for Dutch
patients to contact the GPC. Often they will get self-management
advice (in about 25% of cases).8 Part of these telephone advices will In both countries, women are more likely to seek help from the
GPC than men. This corresponds with the general trend that women
more often use (out-of-hours) primary care.7,8,19,21,23,24 It could also 1022 European Journal of Public Health Figure 2 Age distribution of GPC users and age distribution of total population. Contacts include face-to-face clinic consultations and
telephone consultations. Unit of analysis is patients. Belgium: N=40 152 patients; The Netherlands: N=1 105 932 patients Figure 2 Age distribution of GPC users and age distribution of total population. Contacts include face-to-face clinic consultations and
telephone consultations. Unit of analysis is patients. Main findings and interpretation Belgium: N=40 152 patients; The Netherlands: N=1 105 932 patients Table 2 The ICPC-codes present in the top 10 list of most frequent ICPC codes for face-to-face clinic consultations at the GPC in either
Belgium or the Netherlands: percentage of the total number of face-to-face clinic consultations and position in top 10 (Belgium N = 46 973
contacts; The Netherlands N = 757 607 contacts)
ICPC
Description
Belgium
The Netherlands
%
Nr in top 10a
%
Nr in top 10a
R74
Upper respiratory infection acute
12.73
1
4.47
2
D73
Gastroenteritis presumed infection
4.46
2
1.61
10
R80
Influenza
3.83
3
0.57
–
H71
Acute otitis media/myringitis
3.79
4
2.69
5
R78
Acute bronchitis/bronchiolitis
3.42
5
0.87
–
R76
Tonsillitis acute
3.12
6
1.33
–
U71
Cystitis/urinary infection other
2.92
7
3.70
3
A77
Viral disease, other/NOS
2.77
8
0.45
–
S18
Laceration/cut
2.51
9
6.31
1
D87
Stomach function disorder
2.02
10
0.18
–
L04
Chest symptom/complaint
0.78
–
1.93
7
D06
Abdominal pain localized other
0.18
–
3.24
4
A03
Fever
0.05
–
2.62
6
L12
Hand/finger symptom/complaint
0.05
–
1.71
8
A80
Trauma/injury NOS
0.05
–
1.620.05
9
a: Nr is position of the ICPC code in the top 10 list of most frequent ICPC codes. Note: Excluding telephone consultations, home visits and patients of 75 years and older. Missing ICPC: 4.83% in Belgium and 0.02% in the
Netherlands. Table 2 The ICPC-codes present in the top 10 list of most frequent ICPC codes for face-to-face clinic consultations at the GPC in either
Belgium or the Netherlands: percentage of the total number of face-to-face clinic consultations and position in top 10 (Belgium N = 46 973
contacts; The Netherlands N = 757 607 contacts) a: Nr is position of the ICPC code in the top 10 list of most frequent ICPC codes. Note: Excluding telephone consultations, home visits and patients of 75 years and older. Missing ICPC: 4.83% in Belgium and 0.02% in the
Netherlands. be related to the higher use of the ED by male patients, mostly for
minor trauma.18,25 infections. This is also seen in Belgian general practices during
office hours.27 In the Dutch top 10, many of these infections also
occur, but less frequently, and they are sometimes coded as
symptoms (A03: Fever). Key points The number of consultations in OOH primary care is higher
in the Netherlands than in Belgium. The number of consultations in OOH primary care is higher
in the Netherlands than in Belgium. The types of health problems presented in OOH primary
care differ; consultations in Belgium are mainly related to
infections, while in the Netherlands there are a larger variety
of health problems. By excluding home visits and patients >75 years in both datasets,
we enlarged the comparability. However, with an ageing population
that continuously grows, this group of patients is important and
interesting to study. p
Differences seem to be related to the gatekeeping role of GPs
in the Netherlands and to organizational differences of OOH
care such as telephone triage, medical advice by telephone,
financial thresholds and the number of years of experience
with the system. g
y
Finally, we mainly tried to relate the results to differences in the
organization of OOH primary care. However, other elements of the
broader healthcare system, the health status of the residents and the
national culture could also influence healthcare use. It is known that
there are cultural differences in help-seeking behavior between
countries.30 In the Netherlands, for example, a relatively large
percentage
of
patients
use
the
GPC
for
non-urgent
health
problems.31 This might be associated with the larger total number
of GPC contacts in the Netherlands. It was, however, not possible to
compare the urgency of the contacts between the two countries,
because the lack of triage in Belgian GPCs. y
The results can inform healthcare professionals and policy
makers who are trying to find the most optimal model for
OOH primary care. Conclusions The use of GPCs differs between the Netherlands and Belgium in
terms of number of consultations and types of healthcare problems
presented. Part of the differences may to be related to the strong
primary care system in the Netherlands, with the GP acting as a
gatekeeper to secondary care. Another part may be related to the
organization of the OOH primary care system, such as telephone
triage, medical advice by telephone and financial thresholds. The
results can inform healthcare professionals and policy makers who
are trying to find the most optimal model for OOH primary care, in
the two countries as well as abroad. 9
Colliers A, Bartholomeeusen S, Remmen R, et al. Improving Care And Research
Electronic Data Trust Antwerp (iCAREdata): a research database of linked data on
out-of-hours primary care. BMC Res Notes 2016;9:259. 10 van der Bij S, Khan N, ten Veen P, et al. Improving the quality of EHR recording in
primary care: a data quality feedback tool. J Am Med Inform Assoc 2017;24:81–7. 11 Huibers L, Koetsenruijter J, Grol R, et al. Follow-up after telephone consultations at
out-of-hours primary care. J Am Board Fam Med 2013;26:373–9. 12 Gaakeer MI, van den Brand CL, Gips E, et al. [National developments in Emergency
Departments in the Netherlands: numbers and origins of patients in the period from
2012 to 2015]. Ned Tijdschr Geneeskd 2016;160:D970. 13 Colliers A, Remmen R, Streffer ML, et al. Implementation of a general practitioner
cooperative adjacent to the emergency department of a hospital increases the
caseload for the GPC but not for the emergency department. Acta Clin Belg
2017;72:49–54. 13 Colliers A, Remmen R, Streffer ML, et al. Implementation of a general practitioner
cooperative adjacent to the emergency department of a hospital increases the
caseload for the GPC but not for the emergency department. Acta Clin Belg
2017;72:49–54. Main findings and interpretation The Belgian health care system has a fee-
for service approach, which could lead to a feeling of reciprocity.28,29
In exchange for the payment, the GP feels the need to give
something in return and several patients also expect something in
return, such as a prescription, because of their past experiences.26
Experience with a certain system is the most important factor to
choose the same service the next time. But many infections are
self-limiting and no treatment is necessary. In the Netherlands,
infections are commonly dealt with by giving patients self-care
advice by telephone, and hopefully the patient, when experiencing
the same symptoms in the future, will know what to do. There are differences in the types of health problems presented
between the services in both countries. These differences could be
related to the larger emphasis on primary care in the Netherlands
and frequent co-location of GPCs with an ED, where the GPC treats
a large part of patients that formerly went to the ED.15 In the Dutch
top 10 of health problems, there was a high prevalence of traumata,
such as lacerations and cuts. In Belgium, traumata are mostly
handled at the ED and patients may have learnt to link these types
of problems with this more technical environment.26 This could lead
to a loss of experience and confidence of GPs with these types of
problems, making it more and more a task for secondary care.6,26
The top 10 of diagnoses in Belgian GPCs mostly consists of 1023 Examining differences in out-of-hours primary care use in Belgium and the Netherlands Examining differences in out-of-hours primary care use in Belgium and the Netherlands Implications for research and/or practice Examining differences between countries using information from
electronic health records can lead to an understanding of the rela-
tionship between the organization of healthcare and healthcare util-
ization. In future studies, more countries could be compared and
data from >1 year could be included to examine trends in healthcare
use. It would also be interesting to make an international
comparison of motives and expectations of patients with regard to
contacting the GPC. 2
Leibowitz R, Day S, Dunt D. A systematic review of the effect of different models of
after-hours primary medical care services on clinical outcome, medical workload,
and patient and GP satisfaction. Fam Pract 2003;20:311–7. 3
Huibers L, Philips H, Giesen P, et al. EurOOHnet-the European research network
for out-of-hours primary health care. Eur J Gen Pract 2014;20:229–32. 4
Huibers L, Smits M, Christensen MB, et al. [General practitioner cooperatives in the
Netherlands and Denmark: a comparison]. Tijdschr Gezondheidswet 2017;95:254–7. 5
Smits M, Rutten M, Keizer E, et al. The development and performance of after-
hours primary care in the Netherlands: a narrative review. Ann Intern Med
2017;166:737–42. Most of our findings are explained through hypotheses. It is not
clear if there are causal relationships and it is likely that some factors
interlock. Future research can further develop, confirm or reject
these hypotheses and assess their transferability to other contexts
or countries. For example, Belgian GPCs are currently implementing
telephone triage. This could have an impact on patient flows and
help seeking behavior and will be of interest for future research. 6
Philips H, Remmen R, Van Royen P, et al. What’s the effect of the implementation
of general practitioner cooperatives on caseload? Prospective intervention study on
primary and secondary care. BMC Health Serv Res 2010;10:222. 7
IMA-AIM Atlas. InterMutualistisch Agentschap, Bruxelles, Belgium. Available at:
http://atlas.ima-aim.be/databanken (6 December 2018, date last accessed). 8
NIVEL Primary Care Database. NIVEL, Utrecht, The Netherlands. Available at: https://
www.nivel.nl/nl/nzr/zorgregistraties-eerstelijn (6 December 2018, date last accessed). References 1
Huibers L, Giesen P, Wensing M, Grol R. Out-of-hours care in western countries:
assessment of different organizational models. BMC Health Serv Res 2009;9:105. 1
Huibers L, Giesen P, Wensing M, Grol R. Out-of-hours care in western countries:
assessment of different organizational models. BMC Health Serv Res 2009;9:105. 1
Huibers L, Giesen P, Wensing M, Grol R. Out-of-hours care in western countries:
assessment of different organizational models. BMC Health Serv Res 2009;9:105. Acknowledgements We used routinely recorded electronic health record data, no extra
effort was asked of the GPs. We had access to a large and reliable
sample in both countries. Comparing the two countries was
complicated by the fact that Dutch GPCs have the option to give
a telephone consultation, whereas all patients are seen by a GP in
Belgian GPCs. We do not know how many patients had a face-to-
face clinic consultation after a telephone consultation for the same
health problem. In addition, for the analyses of ICPC codes, the
threshold for inclusion of GPCs differed between the countries,
based on national quality criteria. Moreover, we noticed differences
in the use of ICPC codes by the Dutch and Belgian GPs, which
decreased comparability. Belgian GPs have the tendency to use
diagnosis codes, while Dutch GPs more often use symptom codes. This could be related to differences in the software that helps the GP
choose a ICPC, in training in coding or in cultural differences,
whereby coding a diagnosis is more solution orientated and gives
less the feeling of uncertainty. We thank all GPCs that participate in NIVEL Primary Care Database
and iCAREdata and we thank EurOOHnet for providing a forum for
collaboration. Funding The iCAREdata database is funded by the Research Foundation
Flanders. Conflicts of interest: None declared. Supplementary data Supplementary data are available at EURPUB online. Supplementary data are available at EURPUB online. 1024 European Journal of Public Health 14 Ramlakhan S, Mason S, O’Keeffe C, et al. Primary care services located with EDs: a
review of effectiveness. Emerg Med J 2016;33:495–503. 24 Turner LR, Pearce C, Borg M, et al. Characteristics of patients presenting to an
after-hours clinic: results of a MAGNET analysis. Aust J Prim Health
2017;23:294–9. 15 Rutten M, Vrielink F, Smits M, Giesen P. Patient and care characteristics of self-
referrals treated by the general practitioner cooperative at emergency-care-access-
points in the Netherlands. BMC Fam Pract 2017;18:62. 25 de Valk J, Taal EM, Nijhoff MS, et al. Self-referred patients at the Emergency
Department: patient characteristics, motivations, and willingness to make a
copayment (vol 7, 30, 2014). Int J Emerg Med 2015;8. 16 Avalosse H, Chapelle A, van Sloten F. [Resort to the emergency department of a
hospital: exploration of CM data]. CM Informatie 2015;262:18–25. 26 Philips H, Mahr D, Remmen R, et al. Experience: the most critical factor in choosing
after-hours medical care. Qual Saf Health Care 2010;19:e3. 17 de Bont EG, Loonen N, Hendrix DA, et al. Childhood fever: a qualitative study on
parents’ expectations and experiences during general practice out-of-hours care
consultations. BMC Fam Pract 2015;16:131. 27 Truyers C, Goderis G, Dewitte H, et al. The Intego database: background, methods
and basic results of a Flemish general practice-based continuous morbidity regis-
tration project. BMC Med Inform Decis Mak 2014;14:48. 18 Philips H, Remmen R, De Paepe P, et al. Out of hours care: a profile analysis of
patients attending the emergency department and the general practitioner on call. BMC Fam Pract 2010;11:88. 28 Brookes-Howell L, Hood K, Cooper L, et al. Understanding variation in primary
medical care: a nine-country qualitative study of clinicians’ accounts of the non-
clinical factors that shape antibiotic prescribing decisions for lower respiratory tract
infection. BMJ Open 2012;2:e000796. 19 Kelly SJ, Piercy H, Ibbotson R, Fowler Davis SV. Who attends out-of-hours general
practice appointments? Analysis of a patient cohort accessing new out-of-hours
units. BMJ Open 2018;8:e020308. 29 Colliers A, Coenen S, Remmen R, et al. How do general practitioners and
pharmacists experience antibiotic use in out-of-hours primary care? An exploratory
qualitative interview study to inform a participatory action research project. BMJ
Open 2018;0:e023154. 20 Smits M, Peters Y, Broers S, et al. Supplementary data Association between general practice character-
istics and use of out-of-hours GP cooperatives. BMC Fam Pract 2015;16:52. 21 Buja A, Toffanin R, Rigon S, et al. Out-of-hours primary care services: demands and
patient referral patterns in a Veneto region (Italy) Local Health Authority. Health
Policy 2015;119:437–46. 30 Huibers L, Keizer E, Carlsen A, et al. Help-seeking behaviour outside office hours in
Denmark, the Netherlands, and Switzerland: a questionnaire study exploring
responses to hypothetical cases. BMJ Open 2018;8:e019295. 22 Karen Voigt K, Tache´ S, Klement A, et al. Gaining information about home visits in
primary care: methodological issues from a feasibility study. BMC Fam Pract 2014;15:87. 31 Keizer E, Smits M, Peters Y, et al. Contacts with out-of-hours primary care for
nonurgent problems: patients’ beliefs or deficiencies in healthcare?. BMC Fam Pract
2015;16:157. 23 Huber CA, Rosemann T, Zoller M, et al. Out-of-hours demand in primary care:
frequency, mode of contact and reasons for encounter in Switzerland. J Eval Clin
Pract 2011;17:174–9. p
f
The Author(s) 2019. Published by Oxford University Press on behalf of the European Public Health Association. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/
4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. doi:10.1093/eurpub/ckz081
Advance Access published on 14 May 2019 Ambulatory care-sensitive emergency department
cases: a mixed methods approach to systemize and
analyze cases in Germany Wiebke Schuettig, Leonie Sundmacher Department of Health Services Management, Ludwig Maximilians University Munich, Munich, Germany Department of Health Services Management, Ludwig Maximilians University Munich, Munich, Germany Correspondence: Wiebke Schuettig, Department of Health Services Management, Ludwig Maximilians University Munich,
Schackstrasse 4, 80539 Munich, Germany, Tel: +49 (0) 8921801347, Fax: +49 89 2180 99 1347, e-mail address:
schuettig@bwl.lmu.de Background: Internationally, emergency departments (ED) are treating increasing numbers of patients with conditions
that could have been managed appropriately in ambulatory care (AC) settings. The aim of our study was to develop the
first consensus-based list of AC-sensitive conditions commonly seen in German EDs and explore predictors of these visits. Methods: Our study used a Delphi survey of 30 physicians to compile a list of conditions they agreed were amenable to
AC treatment. The group identified reasons why patients visit EDs instead of AC. We used the results to inform spatial
regression models analysing the association of patient characteristics and attributes of AC with AC-sensitive ED visits
based on 2015 district-level data. Results: Our study provides a list of AC-sensitive conditions based on the German
ED context. Results suggest that, up to the age of 70 years, the older the patients, the less likely they seek EDs for
these conditions. Results of our regression analyses suggest that AC-sensitive ED rates were significantly higher in
districts with lower physician density. Patients’ urgency perception and preferences were identified as main drivers
of AC-sensitive ED visits. Conclusion: Future policy measures should aim to help guide patients through the
healthcare system so that they receive the best care in place that is most appropriate in terms of quality, safety
and continuity of care. A list of AC-sensitive ED conditions can be used as a monitoring instrument and for further
analyses of routine data to inform policy makers seeking to improve resource use and allocation.
|
https://openalex.org/W4317728354
|
https://www.mdpi.com/2073-4336/11/1/7/pdf?version=1580033614
|
English
| null |
Acknowledgment to the Reviewers of Games in 2022
|
Games
| 2,023
|
cc-by
| 765
|
Editorial Games Editorial Office
MDPI, St. Alban-Anlage 66, 4052 Basel, Switzerland
Published: 26 January 2020 The editorial team greatly appreciates the reviewers who have dedicated their considerable time
and expertise to the journal’s rigorous editorial process over the past 12 months, regardless of
whether the papers are finally published or not. In 2019, a total of 49 papers were published in the
journal, with a median time to first decision of 30.72 days and a median time from submission to
publication of 73 days. The editors would like to express their sincere gratitude to the following
reviewers for their generous contribution in 2019: Dahm, Matthias
Dalhaus, Tobias
Danese, Giuseppe
DE MARCO, Giuseppe
Deer, Lachlan
Delaney, Jason
Di, Sharon
Dimant, Eugen
Doruk, Iris
Dürsch, Peter
Ellis, Andrew
Erovenko, Igor
Espinosa, Maria Paz
FARVAQUE, Etienne
Fernández-Revelles, Andrés B. Filson, Darren
Fochmann, Martin
Fooken, Jonas
Frąckiewicz, Piotr
Fu, Jingcheng
Fujita, Toshiyuki
Galliera, Arianna
Gangl, Katharina
Ganzfried, Sam
Ghidoni, Riccardo
Goff, Sandra
Gómez-Miñambres, Joaquín
Gonzalez-Ramirez, María Jimena
Gratton, Gabriele
Grimshaw, Shaun B Abdalzaher, Mohamed S. Adriani, Fabrizio
Akhmetzhanov, Andrei R. Aleksandr, Alekseev
Alm, James
Alt, Marius
Alvarez-Bermejo, Jose Antonio
Ansink, Erik
Aragones, Enriqueta
Archetti, Marco
Arieli, Itai
Attanasi, Giuseppe
Attanasi, Giusppe
Aurell, Alexander
Badicu, Georgian
Bartholomae, Florian
Béal, Sylvain
Bilancini, Ennio
Biondo, Alessio Emanuele
Boonen, Tim J. Brownback, Andy
Bruttel, Lisa
Buchen, Clemens
Cartwright, Edward
Casajus, André
Castillo, Geoffrey
Cavalli, Fausto
Chessa, Michela
Coatney, Kalyn
Cunningham, David E. Editorial www.mdpi.com/journal/games Games 2020, 11, 7; doi:10.3390/g11010007 2 of 3 Games 2020, 11, 7
Gu, Junjian
Gueron, Yves
Gürerk, Özgür
Hanappi, Hardy
Hariskos, Wasilios
Harre, Michael
Harrison, Glenn
Hartman, Bruce
Hedges, Jules
Heine, Florian
Heintz, Christophe
Hilbe, Christian
Hochman, Guy
Hod, Shahar
Hoffer, Adam
Holzman, Ron
Howe, Lance
Hoyer, Britta
Huttegger, Simon
Ikerionwu, Charles
Jaffe, Sonia
Jajac, Nikša
Javarone, Marco Alberto
Javarone, Marco Alberto
Jin, Haiming
Johnson, David
Jung, Sungdo
Kawamura, Kohei
Kirstein, Roland
Klazar, Stanislav
Koch, Alexander
Kölle, Felix
Kolokoltsov, Vassili
Koloniari, Georgia
Kramer, Berber
Krockow, Eva
Kroumi, Dhaker
Kupferman, Orna
Kurokawa, Shun
Kurschilgen, Michael
Lanotte, Ruggero
Lee, Jinkwon
Leland, Jonathan Gu, Junjian
Gueron, Yves
Gürerk, Özgür
Hanappi, Hardy
Hariskos, Wasilios
Harre, Michael
Harrison, Glenn
Hartman, Bruce
Hedges, Jules
Heine, Florian
Heintz, Christophe
Hilbe, Christian
Hochman, Guy
Hod, Shahar
Hoffer, Adam
Holzman, Ron
Howe, Lance
Hoyer, Britta
Huttegger, Simon
Ikerionwu, Charles
Jaffe, Sonia
Jajac, Nikša
Javarone, Marco Alberto
Javarone, Marco Alberto
Jin, Haiming
Johnson, David
Jung, Sungdo
Kawamura, Kohei
Kirstein, Roland
Klazar, Stanislav
Koch, Alexander
Kölle, Felix
Kolokoltsov, Vassili
Koloniari, Georgia
Kramer, Berber
Krockow, Eva
Kroumi, Dhaker
Kupferman, Orna
Kurokawa, Shun
Kurschilgen, Michael
Lanotte, Ruggero
Lee, Jinkwon Louis, Philippos
Mantovani, Marco
Manzoni, Elena
Marini, Marco
Marrouch, Walid
Massey, James
Meloni, Sandro
Millner, Edward
Molinero, Xavier
Munger, Michael C. Musau, Andrew
Naumann, Elias
Nax, Heinrich H. Nicolò, Antonio
Nielsen, Kirby
Norman, Thomas W. L. O’Brien, Michael J. Okada, Isamu
Oncioiu, Ionica
Otjes, Simon
Owen, Guillermo
Pancotto, Francesca
Pawlowitsch, Christina
Pieperhoff, Martina
Pin, Paolo
Podviezko, Askoldas
Prummer, Anja
Puppe, Clemens
Rass, Stefan
Reeck, Crystal
Rettieva, Anna
Robertson, Matthew
Rosa Garcia, Alfonso
Samu, Flóra
SCARTASCINI, CARLOS
Schneider, Frédéric
Schurr, Amos
Schwanebeck, Benjamin
Schwieren, Christiane
Sefton, Martin
Serena, Marco
Shan, Xiaojun (Gene) Louis, Philippos
Mantovani, Marco
Manzoni, Elena
Marini, Marco
Marrouch, Walid
Massey, James
Meloni, Sandro
Millner, Edward
Molinero, Xavier
Munger, Michael C. Musau, Andrew
Naumann, Elias
Nax, Heinrich H. Nicolò, Antonio
Nielsen, Kirby
Norman, Thomas W. L. O’Brien, Michael J. Okada, Isamu
Oncioiu, Ionica
Otjes, Simon
Owen, Guillermo
Pancotto, Francesca
Pawlowitsch, Christina
Pieperhoff, Martina
Pin, Paolo
Podviezko, Askoldas
Prummer, Anja
Puppe, Clemens
Rass, Stefan
Reeck, Crystal
Rettieva, Anna
Robertson, Matthew
Rosa Garcia, Alfonso
Samu, Flóra
SCARTASCINI, CARLOS
Schneider, Frédéric
Schurr, Amos
Schwanebeck, Benjamin
Schwieren, Christiane
Sefton, Martin
Serena, Marco
Shan, Xiaojun (Gene)
Silva, Rui Games 2020, 11, 7
3 of 3
Sirghi, Nicoleta
Sopher, Barry
Spraggon, John
Stashko, Allison
Stoddard, Brock
Swope, Kurtis J.
Te Velde, Vera
Tohme, Fernando
Tröger, Thomas
Tyson, Scott
Ueda, Suguru
Van Basshuysen, Philippe
Van Veldhuizen, Roel
Vesperoni, Alberto
Veszteg, Róbert F.
Veszteg, Róbert Ferenc
Vigors, Belinda
Vlassopoulos, Michael
Vyrastekova, Jana
Wang, Chengsi
Yamarik, Steven
Yang, Yongjie
Ye, Min
Zelizer, Adam
Zusai, Dai
© 2020 by the author. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/). 3 of 3 © 2020 by the author. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/). © 2020 by the author. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).
|
https://openalex.org/W3215314392
|
https://www.researchsquare.com/article/rs-1059888/latest.pdf
|
English
| null |
High Efficiency Laser-Driven Proton Sources Using 3D-Printed Micro-Structure
|
Research Square (Research Square)
| 2,021
|
cc-by
| 7,784
|
High Efficiency Laser-Driven Proton Sources Using
3D-Printed Micro-Structure
Chengyu Qin
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Hui Zhang
(
zhanghui1989@siom.ac.cn
)
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Shun Li
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Nengwen Wang
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Angxiao Li
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Lulin Fan
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Xiaoming Lu
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Jinfeng Li
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Rongjie Xu
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences High Efficiency Laser-Driven Proton Sources Using
3D-Printed Micro-Structure
Chengyu Qin
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Hui Zhang
(
zhanghui1989@siom.ac.cn
)
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Shun Li
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Nengwen Wang
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Angxiao Li
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Lulin Fan
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Xiaoming Lu
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Jinfeng Li
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Rongjie Xu
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences Page 1/18
High Efficiency Laser-Driven Proton Sources Using
3D-Printed Micro-Structure
Chengyu Qin
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Hui Zhang
(
zhanghui1989@siom.ac.cn
)
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Shun Li
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Nengwen Wang
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Angxiao Li
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Lulin Fan
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Xiaoming Lu
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Jinfeng Li
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Rongjie Xu
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Cheng Wang
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Xiaoyan Liang
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences High Efficiency Laser-Driven Proton Sources Using
3D-Printed Micro-Structure gy
Q
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
H i Zh
( h
h i1989
i
) Yuxin Leng Yuxin Leng
State Key Laboratory of High Field Laser Physics,Shanghai Institute of Optics and Fine Mechanics,
Chinese Academy of Sciences
Baifei Shen Lulin Fan Page 1/18 Ruxin Li Chinese Acad Sci, Shanghai Inst Opt & Fine Mech, State Key Lab High Field Laser Phys, Shanghai
201800, Peoples R China. Article Keywords: High efficiency, laser-driven proton sources, 3D-printed micro-structure, 3D-printed microwire-
array, energy conversion efficiency, pre-plasma, experimental conditions Posted Date: November 23rd, 2021 DOI: https://doi.org/10.21203/rs.3.rs-1059888/v1 License:
This work is licensed under a Creative Commons Attribution 4.0 International
License. Read Full License Version of Record: A version of this preprint was published at Communications Physics on May 25th,
2022. See the published version at https://doi.org/10.1038/s42005-022-00900-8. Page 2/18 Page 2/18 Page 2/18 Abstract We applied 3D-printed microwire-array (MWA) structure to boost the energy conversion efficiency of laser
proton acceleration. The advanced nano-printing technique allows precise control on the spacing and
geometrical size of 3D structures at 100-500 nm resolution. Under irradiation of high contrast laser pulse
(15J, 35fs), the MWA target generates over 1.2×1012 protons (> 1MeV) with cut-off energies extending to
25MeV, corresponding to top-end of 8.7% energy conversion efficiency from femtosecond lasers. When
comparing to flat foils the efficiency is enhanced by three times, while the cut-off energy is increased by
30-70% depending on their thicknesses. By precisely controlling the array period via 3D nano-printing, we
found the dependence of proton energy/conversion-efficiency on the spacing of the MWA. The
experimental trend is well reproduced by hydrodynamic and Particle-In-Cell simulations, which reveal for
the first time the modulation of pre-plasma profile induced by laser diffraction within the fine structures. Optimal geometry for laser-proton acceleration is therefore strongly modified. Our work validates the use
of 3D-printed micro-structures to produce high efficiency laser-driven particle sources and pointed out the
new effect in optimizing the experimental conditions. Background In the past two decades, laser-driven proton acceleration has been widely studied for its significance in
applications such as probing high energy density states1, treating cancer therapy2, fast ignition fusion3,
laboratory astrophysics4 and so on. These applications impose certain requirements for the energy and
current intensity of proton beams. By optimizing the laser conditions and thickness of planar targets,
protons could be accelerated up to 85 MeV via the robust target normal sheath acceleration (TNSA)
mechanism5-7. A higher cutoff energy of 94 MeV has been reported via radiation pressure-sheath
acceleration in an ultrathin planar foil8. These sub-100MeV protons are obtained by relatively long laser
pulses of 100’s J energies. For femtosecond lasers, typical cutoff energy of protons lies in 30-70 MeV
using planar targets9-11, since the pulse energy is usually much less than picosecond lasers. Realizing the above-mentioned applications of laser-driven proton sources strongly relies on maximizing
the laser-proton energy conversion efficiency. In laser-foil interaction, reducing target thickness12 is
usually adopted to improve laser-to-proton energy conversion efficiency. In general, plasma targets
irradiated by picosecond lasers behave differently from a femtosecond laser because the instability of
the plasma and self-generated electromagnetic fields expand over time13,14. Consequently, high laser-to-
proton energy conversion efficiency up to 10%-15%5,15 tends to appear in TNSA when using large energy
picosecond laser pulses. The efficiency of 3%-4%11,16 is achieved on femtosecond lasers of much lower
pulse energy, which is still below the theoretical value ~8% predicted by Sentoku et al17. For ultra-short laser pulses, optimizing the target condition to enhance the laser absorption efficiency has
become a promising solution. Employing carbon nanotube foam (CNF) on a planar diamondlike carbon
(DLC) foil has realized triple energy gain for protons18,19. An alternative method is to introduce Page 3/18 periodic/aperiodic nano/micro-structures in front of planar targets20-28. Experimental data indicate that
microstructure can greatly raise the temperature of hot electrons beyond that of the ponderomotive
acceleration29,30. In the case of picosecond relativistic lasers irradiating microstructure targets at large
angles, the yield of electron and x-ray could be increased31,32, but the proton energy from TNSA remains
unchanged27 or even becomes less33. The main reason is that the rising edge of picosecond lasers is
long enough to ionize the surface structure so that the plasma infill could shutter the laser from further
interaction. Considerable energy enhancement for protons has been observed by using defocused
picosecond lasers with normal incidence (1017~1018 W/cm2)25. Background The effect of pre-plasma infill is
suppressed when employing ultrashort femtosecond relativistic lasers28. Judging from the reported
results, when using fine surface structures to improve proton acceleration in TNSA, high contrast
femtosecond lasers with an incidence angle as small as possible is more favorable. We notice that the fast development of three-dimensional (3D) nano-printing technique allows for well-
controlled fabrication of fine solid structures. The two-photon polymerization (TPP) method can readily
achieve lateral resolution ~100 nm and vertical resolution ~500 nm34. Compared to other approaches
such as Si semiconductor-based technique33 and electrochemical deposition method28, 3D nano-printing
can accurately print various complex structures as desired in experiments at micro or sub-micro scale. It
is therefore timely to apply this advanced technique to laser-plasma physics such as to manipulate
ultrafast laser-plasma interaction22, generation of high bright x/gamma-ray35, high power coherent
Terahertz36 and positrons37. In this paper, we use the TPP Nanoscribe 3D printer38 to fabricate microwire array (MWA) onto SiN
flat foils and investigate laser-driven proton acceleration at various spacing conditions of the array. The
MWA structure boosts the laser-to-proton energy conversion efficiency to 8.7% (total number 1.2×1012 for
proton energies more than 1 MeV), about three times more than that of flat foils of optimal thickness. After implementing the MWA structure the cut-off energy of protons is increased by 70% compared to flat
foils of similar thickness (4 µm-thickness). It reaches 25 MeV (MWA targets) at on-target laser intensity of
about 2.2×1020 W/cm2, which is 30% more than that of the optimized flat foil (1 µm-thickness). We
change the spacing of MWA precisely via 3D nano-printing and find out how it influences the proton
acceleration process. Our 2D hydrodynamic simulations point out that the pre-plasma condition of MWA
targets induced by the pre-pulse is significantly different from the flat target case where the density length
scale is almost uniform in the latter. The micro-structures can induce strong diffraction of the incident
pre-pulse, resulting in featured pre-plasma distributions depending on the surface profile. After bringing
the pre-expansion plasma conditions into 2D particle-in-cell (PIC) simulations, we reproduce the
experimental results and find the optimal MWA parameters for proton acceleration. Methods MWA fabrication. The 3D printing machine we employed is based on two-photon polymerization
technique34. A brief schematic diagram is shown in Fig. 1(b). A femto-fiber laser system delivers pulses at Page 4/18 Page 4/18 a center wavelength of 780 fs with a repetition rate of 80 MHz. The light pulses typically exhibit a
duration of 100 fs with an average power of more than 350 mW. The laser is guided into the objective
lens with a magnification of 63×(NA1.4) and focused on the sample surface. The photo-resin is partial
exposed following the designed structure model. To remove unexposed photo-resist, the structures are
developed in the developer liquid followed by careful rinsing with isopropanol (IPA). After UV treatment42
and natural drying in air, solid structures are obtained. The structure is a polymer C14H18O7 with a mass
density of ~1.17 g/cm3. We use the commercial 3D printer of Nanoscribe Photonic Professional GT138 to
fabricate MWAs onto the 1-μm SiN substrates within an area of 400 μm × 400 μm. The diameter and
height of microwires were fixed at d=0.4 μm and h=2.5 μm, while their period varied from p=1.3 μm to p
=3.3 μm. Simulations. The hydrodynamics simulations were performed using the 2D FLASH code48 in Cartesian
coordinate system. A flat-top laser with a steep rising edge irradiated the Al MWA at an incident angle of
5.5°. The material initial temperature was set to 290 K. The laser intensity is 5×109 W/cm2 and ablation
time was 500 ps for Figs. 3(a) and (b). The PIC simulation was performed using the 2D EPOCH code49. The simulation box was 65 μm × 30 μm with mesh cell size equal to 4 nm × 8 nm. The incoming laser
pulse temporal and spatial profiles were both Gaussian, with FWHM equal to 35 fs and 10 μm, Simulations. The hydrodynamics simulations were performed using the 2D FLASH code48 in Cartesian
coordinate system. A flat-top laser with a steep rising edge irradiated the Al MWA at an incident angle of
5.5°. The material initial temperature was set to 290 K. The laser intensity is 5×109 W/cm2 and ablation
time was 500 ps for Figs. 3(a) and (b). The PIC simulation was performed using the 2D EPOCH code49. The simulation box was 65 μm × 30 μm with mesh cell size equal to 4 nm × 8 nm. Methods The incoming laser
pulse temporal and spatial profiles were both Gaussian, with FWHM equal to 35 fs and 10 μm,
respectively. The peak intensity was 2.2×1020 W/cm2. Open boundary condition was used in all
simulations. The flat targets were initialized as a uniform mixture of Si14+ and N7+, both at 12 nc. The
flats coupled with CH contaminant layers at the rear of the film with thickness of 20 nm and electron
density of 45 nc. For MWA targets, the wires were defined as fully ionized CH and the electron density
were 210 nc (mass density ~ 1.17 g/cm3). Each pillar is surrounded with modulated pre-plasma of two
components. Around the tip the pre-plasma forms a semicircular with npre1 = ne1exp(-△r/lh), where
ne1, △r, and lh are the pillar electron density, the distance to the tip center and the corresponding density
scale length, respectively. The pre-plasma density in the rest area takes the form npre2= ne1exp[-
(△y/lh +△x/lx)]+ne2exp(-△ /lf), where ne2,△(x ,y), lx,f ,is the foil electron density, the distance to initial
target boundary and the related scale lengths, respectively. The substrate was the same as the flat
situation. Experimental Results Experimental set-up. The experiments were carried out in a petawatt-class Ti:sapphire laser system with
central wavelength λ = 800 nm, which delivers around 15±2 J on-target energy within 35 fs duration (full
width at half maximum, FWHM) in the current run. About 40% of the laser energy is enclosed in a focal
spot of 10 μm diameter (FWHM), yielding a peak intensity of about (2.2±0.3)×1020 W/cm2. The laser
contrast of amplified spontaneous emission (ASE) pedestal is ~1011 (1010) at 30 (6) ps prior to the main
pulse39, which could induce pre-plasmas with typical scale length of several hundreds of nanometers40. Our micro-structures of MWA targets used here survived the above pre-pulse condition. The sketch of the
experimental setup is shown in Fig. 1(a). The p-polarized laser pulse is focused by a f / 4 off-axis Page 5/18 parabolic mirror onto the target at an incident angle of 5.5°, which is the smallest angle one can set to
avoid light backscattering to the laser chain. A stack of radiochromic films (RCFs) with a 3-mm-diameter
hole in the center are enwrapped with 15 μm-thickness Al foil to shield the stack from debris. It is located
at L1 = 5.3 cm to measure the proton energy spectrum and profile. Several EBT3-type RCFs of high
sensitivity are placed behind the HD-V2s in stack to guarantee the accuracy near the spectral cut-off. The
ion transport code SRIM41 was used to model the range of proton energies stopped in each layer. Following that a Thomson parabola (TP) spectrometer equipped with a BAS-TR image plate is set at L2 =
46 cm. Both of RCF stacks and TP are aiming at the target normal direction. We fix the diameter and height of the micro-wires of MWA targets at d=0.4 μm and h=2.5 μm and vary the
period, i.e., p = 1.3 μm, 1.8 μm, 2.3 μm and 3.3 μm. The diagram of laser direct writing (see Methods)
employed in our experiment is sketched in Figure 1(b). Figures 1(c) and (d) show the scanning electron
microscope (SEM) images of two typical printed MWA structures with p = 1.3 μm and 3.3 μm,
respectively. The MWAs are printed onto 1 μm-thickness (the optimal thickness of flat foils for TNSA
under the current laser condition) SiN foil, covering an area of about 400 μm × 400 μm. Experimental Results This is much
greater than the laser focal spot such that the laser beam could hit the MWA considering the beam jitter. The printed material is polymer with a mass density of 1.17 g/cm3. In addition, ultra violet (UV) treatment
is employed before they are dried to maintain the stability of the structures42. Enhancing proton acceleration. At certain pre-pulse condition there exists an optimal thickness of planar
targets for TNSA43. To identify the enhancement brought by the micro-structure, it is very important to
exclude the effect of target thickening due to the additional structure. Therefore, before doing comparison
we first find out the optimal foil thickness that gives highest proton energy. This was omitted in previous
reports26-28, where the structure target is compared to the planar target with the same thickness of only
the substrate, or the structure (substrate thickness is ignored). We scan over the thickness of planar SiN
layers from 0.2 μm to 4.0 μm and summarize the measured results in Fig. 2(a) (black square). It can be
clearly seen that with the target thickness increasing, the cut-off energy of protons increases first and
then declines. For 1 μm-thickness flat foils, the proton energy reaches maximum of 18~19 MeV. For the case of MWA targets, the substrate thickness remains unchanged at 1 μm. Figure 2(a) also shows
the measured cutoff energy of protons as a function of the microwire period of MWA targets (red circle). It should be noted that the cut-off energy for all period conditions of MWA targets are above the highest
obtained with flat foils. As the array period increases from 1.3 μm to 3.3 μm, the cut-off energy of protons
rises from 20 MeV to 25 MeV. The maximum energy gain of protons is experimentally observed for MWA
targets of 3.3 μm period, which is 30% more than that with 1 μm SiN-foil. The combined substrate and
structure correspond to an effective thickness of about 4 μm for flat SiN-foil. When comparing to the
latter, the MWA energy enhancement is close to 70%. Representative proton spectra obtained from RCF stacks over the whole proton beam and TP
spectrometer for 1 μm SiN foils, and MWA targets with three periods of 1.3 μm, 2.3 μm, and 3.3 μm are
shown in Fig. 2(c). The absolute response of the RCF dosimeter was calibrated following the method Page 6/18 introduced previously44,45. Experimental Results The experimental results of proton spectra from RCF stacks are consistent
with that of TP spectrometer. The total number of protons for MWA targets of 3.3 μm period with energy
more than 1MeV is about 1.2×1012 according to RCF spectrum integral, approximately 2.5 times more
than that of 1 μm SiN foils of 4.8×1011. Here, the Al layer with thickness of 15 μm could minimize
possible dose contribution from other heavy ion species. The presence of this foil limited the minimum
detectable proton energy to around 1 MeV. Further, the laser-to-proton energy conversion efficiency (η) is summarized in Fig. 2(b). As the foil
thickness decreases from 4 μm to 200 nm, a clear increase in conversion efficiency is observed. The
number of 4.5% obtained with 200 nm flat can be interpreted as the appearance of piston-like
acceleration due to the initial density gradient of rear pre-plasma46,47. To exclude this complication, we
limit our discussion in the TNSA dominated regime (thickness ). In this case, the maximum η of planar
SiN is about 2.9±0.2% for 1 μm SiN foils. When equipping MWA on 1 μm SiN foils, the η is significantly
enhanced. The efficiency exhibits similar variation tendency as the cutoff energy such that the
η increases with larger array spacing. The maximum η of MWA target reaches to 8.7% for one shot at
p=3.3 μm, corresponding to a total of 1.1 J for protons with energy > 1 MeV. This is three times that of the
average value for the 1 μm SiN foil thickness. This high conversion efficiency is at the top end of report
values for TNSA driven by Ti:sapphire based 10’s J laser systems11,16, which exceeds the maximum
conversion efficiency predicted in the Ref. [30]. It should be noted that even though the cut-off energy for
1.3 μm MWA targets is almost the same as that of the 1 μm SiN, the energy conversion efficiency is
almost doubled for the former. Both the cut-off energy and conversion efficiency increase with larger
spatial period, indicating that the acceleration does not reach maximum for the MWA periods chosen in
the current run. We therefore perform further simulations to interpret the experimental observation and
more importantly, to find out the optimal condition for laser-proton acceleration. Simulation And Discussion To better understand the experimental results, we carry out hydrodynamic and PIC simulations. The pre-
plasma distribution is determined by 2D hydrodynamic simulations in Cartesian coordinate system using
the FLASH48 code. Limited by the available material in the code, the Al material data is used in numerical
simulation. The 3D-printed-MWA parameters are inputted into FLASH. After the pre-pulse ablating MWA
targets, it can be seen from Fig. 3(b) that the heads of the pillars expand more obviously than their roots. The plasma forms large clouds around the tips, which narrows the gap between the wires. This type of
plasma expansion is more severe for denser pillar arrays. It may even shutter the laser from further
interaction as shown in Fig. 3(a). Figs. 3 (c) and (d) show the mass density distribution along three y-
positions y= 17 μm, 18 μm and 19 μm from Figs. 3 (a) and (b), corresponding to the inside, edge and
outside of the array respectively. Obvious blocking inside the array can be seen in Fig. 3(c), while the
structure remains open for arrays with larger gaps (p=3.3 μm) in Fig. 3 (d). Page 7/18 We further show the laser field distribution when incident onto the MWA structure. As seen in Fig. 3(e), the
laser field is diffracted by the periodic structure such that it is locally intensified around the tips of the
pillars. Modulated laser intensity produces pre-plasma of larger scale length in the light-intensified
vicinity, leading to the featured plasma profile in Figs. 3(a) and (b) Based on the results of FLASH simulations, we set the density distribution of pre-expanding MWA targets
in Fig. 3(f). Each pillar is surrounded with modulated pre-plasma of two components. Around the tip the
pre-plasma forms a semicircular with npre1 = ne1exp(-△r/lh), where ne1, △r, and lh are the pillar electron
density, the distance to the tip center and the corresponding density scale length, respectively. The pre-
plasma density in the rest area takes the form npre2 = ne1exp[-(△y/lh + △x/lx)]+ne2exp(-△ /lf), where
ne2, △(x ,y), lx,f , is the foil electron density, the distance to initial target boundary and the related scale
lengths, respectively. We take the above modulated pre-plasma distribution in the following 2D PIC simulations using the code
EPOCH49. The simulation box size was Wx × Wy = 65 μm × 30 μm with the cell size of dx = 4 and dy = 8
nm. Simulation And Discussion The front surface of flat target is located at x=0, with pre-
plasma of scale length lf = 100 nm. We vary the foil thickness between 0.2 μm, 1 μm, 2 μm and 4 μm. In
particular, for the ultrathin 0.2 μm foil a rear pre-plasma with the same scale length is also added 46. The
related simulation results of cut-off energy and spectra of protons are summarized in Figs. 2(a) and (d),
which are in reasonable agreement with the experimental results. The pre-plasma parameters for MWA targets employed in 2D PIC simulations are based on the
hydrodynamic results as shown in Fig. 3(f). For spatial period of 2.3 and 3.3 μm, we set lh = 350 nm and
lx = 1 μm while the one of the flat foil is lf = 100 nm, according to the modulated pre-pulse intensity by the
periodic structure. At small period of 1.3 μm, lh is set to 100 nm according to the distribution in Fig. 3(a). Here the wires are defined as fully ionized CH and the electron density is 210 nc (mass density ~ 1.17
g/cm3). Page 8/18
From the electron energy spectra in Fig. 4(a) it is seen that the MWA generates much higher temperature
due to the DLA mechanism50. Here the highest temperature appears at p=5.3 μm, which was not tested in
the current experimental run. The corresponding temporal evolution of the longitudinal electric field Ex are
plotted in Fig. 4(b). We see that the peak field strength is largely enhanced due to the MWA. For the
spatial period shown here (3.3 μm), one finds higher peak value for the situation without pre-plasma while
a slow decrease with pre-plasma, indicating stronger proton acceleration for the former. As pointed out in Page 8/18
From the electron energy spectra in Fig. 4(a) it is seen that the MWA generates much higher temperature
due to the DLA mechanism50. Here the highest temperature appears at p=5.3 μm, which was not tested in
the current experimental run. The corresponding temporal evolution of the longitudinal electric field Ex are
plotted in Fig. 4(b). We see that the peak field strength is largely enhanced due to the MWA. For the
spatial period shown here (3.3 μm), one finds higher peak value for the situation without pre-plasma while
a slow decrease with pre-plasma, indicating stronger proton acceleration for the former. Simulation And Discussion Particle-per-cell is set to 9. The p-polarized Gaussian laser pulse is incident at 5.5° from the left side
with a central wavelength of 800 nm of pulse duration of 35fs (FWHM). Its peak intensity reaches 2.2 ×
1020 W/cm2 after being focused to 10 μm spot size. There parameters are the same as experimental
conditions. The targets are fully ionized as cold plasma and keep electrically neutral. The flat targets
consist of Si14+ and N7+ with the same density of 12 nc (total electron density is ne =
252nc, nc = meω2/4 e2 is the critical density). A thin CH contaminant layer of 20 nm and electron density
45 nc is attached to the rear side of the foil. The front surface of flat target is located at x=0, with pre-
plasma of scale length lf = 100 nm. We vary the foil thickness between 0.2 μm, 1 μm, 2 μm and 4 μm. In
particular, for the ultrathin 0.2 μm foil a rear pre-plasma with the same scale length is also added 46. The
related simulation results of cut-off energy and spectra of protons are summarized in Figs. 2(a) and (d),
which are in reasonable agreement with the experimental results. We take the above modulated pre-plasma distribution in the following 2D PIC simulations using the code
EPOCH49. The simulation box size was Wx × Wy = 65 μm × 30 μm with the cell size of dx = 4 and dy = 8
nm. Particle-per-cell is set to 9. The p-polarized Gaussian laser pulse is incident at 5.5° from the left side
with a central wavelength of 800 nm of pulse duration of 35fs (FWHM). Its peak intensity reaches 2.2 ×
1020 W/cm2 after being focused to 10 μm spot size. There parameters are the same as experimental
conditions. The targets are fully ionized as cold plasma and keep electrically neutral. The flat targets
consist of Si14+ and N7+ with the same density of 12 nc (total electron density is ne =
252n
n = m ω2/4 e2 is the critical density) A thin CH contaminant layer of 20 nm and electron density 252nc, nc = meω2/4 e2 is the critical density). A thin CH contaminant layer of 20 nm and electron density
45 nc is attached to the rear side of the foil. Simulation And Discussion As pointed out in Page 8/18 Page 8/18 previous studies27,29,30,51, enhancement of the sheath field results from the higher population of energetic
electrons generated in micro-structures. The scale lengths of lh and lx have different influence on the proton maximum energy, as illustrated in Fig. 4(c), where lf is fixed to 100 nm. When changing lx from 0.5 μm to 5 μm with lh = 350 nm, the proton cut-
off energy remains at 27-28 MeV. The scale length of lf has similar effects when changing within several
hundred nanometers range (not shown here). However, the maximum energies drop from 35 MeV to 22
MeV when slightly increasing lh from 250 nm to 450 nm with lf = 1 μm. It suggests that proton cut-off
energy is more sensitive to the scale length of the wire tip lh, which is closely related to the modulated
plasma expansion resulting from pulse diffraction. In other words, the laser contrast has a significant impact on proton acceleration when using MWA
targets. Figure. 4(d) shows proton maximum energy as a function of the wire period with and without pre-
plasma. When the pre-plasma is absent, proton energy is maximized to 42 MeV at optimal period around
2.3 μm. However, when introducing the modulated pre-plasma, the energy distribution shows slightly
lower peak value of 37 MeV at much larger spatial period 5.3-6.3 μm. This trend agrees well with the
electron temperature in Fig. 4(b) and more importantly, with the experimental observation. Because of the
modulated pre-plasma in MWA, the optimal period for proton acceleration is modified such that one
should increase the spatial period as compared to the case when no pre-plasma is considered. However,
there is also tradeoff of using MWA with larger spacing. For finite laser spot size, decrease of wire
number covered by the laser beam may lead to energy drop for protons 28. In our case, optimized laser-
proton acceleration is achieved when the laser spot size covers 0.7~1.5 unit. In the current experimental
run, we had best results at p=3.3 μm. In addition, the energy conversion efficiency from simulations is plotted in Fig. 4(d) after including the
pre-plasma condition. Since the simulations are 2D, we calibrate the efficiency with the average value of
6.9 % measured in experiment for p=3.3 μm, which gives a fixed ratio for all simulation data. The
efficiency shows similar trend as the cut-off proton energy. Simulation And Discussion The highest efficiency appears for p=5.3 μm. When using the maximum calibration factor (8.7% for p=3.3 μm measured in experiment), the η could
boost to 10.7% with p=5.3 μm (not shown here). Snapshots of longitudinal electric field Ex at t=100 fs from simulations are shown in Figs. 5(a-c)
respectively in three cases. According to comparison, it is observed that the field strength and length
scale are both evidently larger for MWA targets. When introducing pre-expanding plasma, much of the
main laser pulse is reflected while only part of the laser pulse could propagate into structure, as shown in
Fig. 5(b). In this case, direct laser acceleration (DLA) of electrons becomes less effective. The latter is
usually expected in micro-wires/tubes to greatly improve electron temperature29,30,51. Comparing to the
case without pre-plasma (Fig. 5(c)), the peak acceleration field is notably smaller in Fig. 5(b). These
distributions agree with the comparison in Fig. 4(d) for p=3.3 μm. We notice the periodic structure induces
diffraction for the main laser pulse in both cases. The reflected laser field is diffracted along large Page 9/18 Page 9/18 divergence angles while the sheath field is also modulated with significant periodic and divergent
patterns. The difference between each case is also imprinted onto the proton acceleration process, as
shown in Figs. 5(d-f), where the one without pre-plasma generates several streams of protons following
each unit of micro-wire. We did not see the streamed profile in the current experimental run due to the pre-
pulse condition here. This feature could be further measured as an identification of the clean micro-
structures during interaction, with higher beam contrasts enabled by plasma mirrors. All relevant data are available from the corresponding authors upon reasonable request. All relevant data are available from the corresponding authors upon reasonable request. Conclusion In conclusion, using a 3D-nano printer to fabricate MWA structures onto the front surface of flat foils, we
have experimentally demonstrated simultaneous increase of the proton maximum energy and laser-
proton energy conversion efficiency. We identified the effect of pre-plasma modulation caused by the
diffraction of laser pre-pulse in MWA structure, which successfully interprets the experimental results. This is the first time to reveal the role of the scale length lh of the tip of wire structures, which depends on
the wire gap and laser pre-pulse. Adjusting the wire number in the laser focal spot and the wire gap could
further improve proton cutoff energy. References 1 Romagnani, L. et al. Dynamics of electric fields driving the laser acceleration of multi-MeV protons. Phys. Rev. Lett.95, 195001 (2005). 1 Romagnani, L. et al. Dynamics of electric fields driving the laser acceleration of multi-MeV protons. Phys. Rev. Lett.95, 195001 (2005). 2 Bulanov, S. V., Esirkepov, T. Z., Khoroshkov, V. S., Kuznetsov, A. V. & Pegorarod, F. Oncological
hadrontherapy with laser ion accelerators. Phys. Lett. A 299, 240-270 (2002). 3 Roth, M. et al. Fast ignition by intense laser-accelerated proton beams. Phys. Rev. Lett.86, 436-439
(2001). 4 Remington, B. A. High energy density laboratory astrophysics. Plasma Phys. Control. Fusion47, A191-
A203 (2005). 5 Wagner, F. et al. Maximum Proton Energy above 85 MeV from the Relativistic Interaction of Laser
Pulses with Micrometer Thick CH2 Targets. Phys. Rev. Lett.116, 205002 (2016). 6 Snavely, R. A. et al. Intense High-Energy Proton Beams from Petawatt-Laser Irradiation of Solids. Phy. Rev. Let.85, 2945 (2000). Page 10/18 7 Hatchett, S. P. et al. Electron, photon, and ion beams from the relativistic interaction of Petawatt laser
pulses with solid targets. Phys. Plasmas7, 2076-2082 (2000). 8 Higginson, A. et al. Near-100 MeV protons via a laser-driven transparency-enhanced hybrid acceleration
scheme. Nat. Commun.9, 724 (2018). 9 Ogura, K. et al. Proton acceleration to 40 MeV using a high intensity, high contrast optical parametric
chirped-pulse amplification/Ti:sapphire hybrid laser system. Opt. Lett.37, 2868-2870 (2012). 10 Ziegler, T. et al. Proton beam quality enhancement by spectral phase control of a PW-class laser
system. Sci. Rep.11, 7338 (2021). 11 Green, J. S. et al. High efficiency proton beam generation through target thickness control in
femtosecond laser-plasma interactions. Appl. Phys. Lett.104, 214101 (2014). 12 Mackinnon, A. J. et al. Enhancement of proton acceleration by hot-electron recirculation in thin foils
irradiated by ultraintense laser pulses. Phys. Rev. Lett.88, 215006 (2002). 13 Sorokovikova, A. et al. Generation of Superponderomotive Electrons in Multipicosecond Interactions of
Kilojoule Laser Beams with Solid-Density Plasmas. Phys. Rev. Lett.116, 155001 (2016). 14 Iwata, N., Kojima, S., Sentoku, Y., Hata, M. & Mima, K. Plasma density limits for hole boring by intense
laser pulses. Nat. Commun.9, 623 (2018). 15 Brenner, C. M. et al. High energy conversion efficiency in laserproton acceleration by controlling
laserenergy deposition onto thin foil targets. Appl. Phys. Lett.104, 081123 (2014). 16 Nishiuchi, M. et al. Efficient production of a collimated MeV proton beam from a polyimide target
driven by an intense femtosecond laser pulse. Phys. References Plasmas15, 053104 (2008). 17 Sentoku, Y., Cowan, T. E., Kemp, A. & Ruhl, H. High energy proton acceleration in interaction of short
laser pulse with dense plasma target. Phys. Plasmas10, 2009-2015 (2003). 18 Bin, J. H. et al. Enhanced Laser-Driven Ion Acceleration by Superponderomotive Electrons Generated
from Near-Critical-Density Plasma. Phys. Rev. Lett.120, 074801 (2018). 19 Ma, W. J. et al. Laser Acceleration of Highly Energetic Carbon Ions Using a Double-Layer Target
Composed of Slightly Underdense Plasma and Ultrathin Foil. Phys. Rev. Lett.122, 014803 (2019). 20 Margarone, D. et al. Laser-driven proton acceleration enhancement by nanostructured foils. Phys. Rev. Lett.109, 234801 (2012). 21 Ceccotti, T. et al. Evidence of resonant surface-wave excitation in the relativistic regime through
measurements of proton acceleration from grating targets. Phys. Rev. Lett.111, 185001 (2013). Page 11/18 Page 11/18 Page 11/18 Page 11/18 22 Ji, L. L., Snyder, J., Pukhov, A., Freeman, R. R. & Akli, K. U. Towards manipulating relativistic laser
pulses with micro-tube plasma lenses. Sci. Rep.6, 23256 (2016). 23 Floquet, V. et al. Micro-sphere layered targets efficiency in laser driven proton acceleration. J. Appl. Phys.114, 083305 (2013).. 24 Blanco, M., Flores-Arias, M. T., Ruiz, C. & Vranic, M. Table-top laser-based proton acceleration in
nanostructured targets. New J. Phys.19, 033004 (2017). 25 Khaghani, D. et al. Enhancing laser-driven proton acceleration by using micro-pillar arrays at high drive
energy. Sci. Rep.7, 11366 (2017). 26 Dozières, M. et al. Optimization of laser-nanowire target interaction to increase the proton acceleration
efficiency. Plasma Phys. Control. Fusion61, 065016(8pp) (2019). 27 Bailly-Grandvaux, M. et al. Ion acceleration from microstructured targets irradiated by high-intensity
picosecond laser pulses. Phys. Rev. E102, 021201(R) (2020). 28 Vallieres, S. et al. Enhanced laser-driven proton acceleration using nanowire targets. Sci. Rep.11, 2226
(2021). 29 Jiang, S. et al. Microengineering Laser Plasma Interactions at Relativistic Intensities. Phys. Rev. Lett. 116, 085002 (2016). 30 Snyder, J. et al. Relativistic laser driven electron accelerator using micro-channel plasma targets. Phys. Plasmas26, 033110 (2019). 31 Moreau, A. et al. Enhanced electron acceleration in aligned nanowire arrays irradiated at highly
relativistic intensities. Plasma Phys. Control. Fusion62, 014013 (9pp) (2020). 32 Shou, Y. et al. High-efficiency water-window x-ray generation from nanowire array targets irradiated
with femtosecond laser pulses. Opt. Express29, 5427-5436 (2021). 33 Sedov, M. V. et al. Features of the generation of fast particles from microstructured targets irradiated
by high intensity, picosecond laser pulses. Laser and Particle Beams37, 176-183 (2019). 34 Maruo, S. References & Fourkas, J. T. Recent progress in multiphoton microfabrication. Laser & Photon. Rev.2,
100-111 (2008). 35 Yi, L., Pukhov, A., Luu-Thanh, P. & Shen, B. Bright X-Ray Source from a Laser-Driven Microplasma
Waveguide. Phys. Rev. Lett.116, 115001 (2016). 36 Yi, L. & Fulop, T. Coherent Diffraction Radiation of Relativistic Terahertz Pulses from a Laser-Driven
Microplasma Waveguide. Phys. Rev. Lett.123, 094801 (2019). Page 12/18 Page 12/18 37 iang, S. et al. Enhancing positron production using front surface target structures. Appl. Phys. Lett.118,
094101 (2021). 37 iang, S. et al. Enhancing positron production using front surface target structures. Appl. Phys. Lett.118,
094101 (2021). 37 iang, S. et al. Enhancing positron production using front surface target structures. Appl. Phys. Lett.118,
094101 (2021). 38 Nanoscribe. https://www.nanoscribe.com/en/. 38 Nanoscribe. https://www.nanoscribe.com/en/. 38 Nanoscribe. https://www.nanoscribe.com/en/. 39 Lu, X., Zhang, H., Li, J. & Leng, Y. Reducing temporal pedestal in a Ti:sapphire chirped-pulse
amplification system by using a stretcher based on two concave mirrors. Opt. Lett.46, 5320 (2021). 40 Gizzi, L. A. et al. Intense proton acceleration in ultrarelativistic interaction with nanochannels. Phys. Rev. Research2, 033451 (2020). 41 Ziegler, J. F., Ziegler, M. D. & Biersack, J. P. The stopping and range of ions in matter (2010). Nucl. Instrum. Methods Phys. Res. Sect. B268, 1818 (2010). 42 Purtov, J., Verch, A., Rogin, P. & Hensel, R. Improved development procedure to enhance the stability of
microstructures created by two-photon polymerization. Microelectron. Eng.194, 45-50 (2018). 43 Kaluza, M. et al. Influence of the laser prepulse on proton acceleration in thin-foil experiments. Phys. Rev. Lett .93, 045003 (2004). 44 Bin, J. H. et al. Absolute calibration of GafChromic film for very high flux laser driven ion beams. Rev. Sci. Instrum.90, 053301 (2019). 45 Chen, S. N. et al. Absolute dosimetric characterization of Gafchromic EBT3 and HDv2 films using
commercial flat-bed scanners and evaluation of the scanner response function variability. Rev. Sci. Instrum.87, 073301 (2016). 46 Green, J. S. et al. High efficiency proton beam generation through target thickness control in
femtosecond laser-plasma interactions. Appl. Phys. Lett.104, 214101 (2014). 47 Grismayer, T. & Mora, P. Influence of a finite initial ion density gradient on plasma expansion into a
vacuum. Phys. Plasmas13, 032103 (2006). 48 Fryxell, B. et al. FLASH: An adaptive mesh hydrodynamics code for modeling astrophysical
thermonuclear flashes. The Astrophys. J. Supplement Series131, 273 (2000). 49 Arber, T. D. et al. Contemporary particle-in-cell approach to laser plasma modelling. Plasma Phys. Control. Fusion 57,113001 (2015). 50 Pukhov, A., Sheng, Z. M. & Meyer-ter-Vehn, J. Particle acceleration in relativistic laser channels. Phys. Plasmas6, 2847-2854 (1999). 51 Ji, L., Jiang, S., Pukhov, A., Freeman, R. & Akli, K. Exploring novel target structures for manipulating
relativistic laser–plasma interaction. High Power Laser Sci. Eng.5, e14 (2017). Page 13/18 Page 13/18 Figures Figure 1
(a) Sketch of the experimental setup. (b) Schematic diagram of laser direct writing that prepares the MWA
structure: the laser pulses with wavelength of 780 nm and duration of 100 fs are controlled via the
galvanometer system to scan space voxel in the photoresin after the objective lens and the laser pulse
energy is modulated by the acousto optic modulator (AOM). 38 Nanoscribe. https://www.nanoscribe.com/en/. (c) and (d) show the SEM images of MWA
with period of 1.3 μm and 3.3 μm, respectively. Figure 1 (a) Sketch of the experimental setup. (b) Schematic diagram of laser direct writing that prepares the MWA
structure: the laser pulses with wavelength of 780 nm and duration of 100 fs are controlled via the
galvanometer system to scan space voxel in the photoresin after the objective lens and the laser pulse
energy is modulated by the acousto optic modulator (AOM). (c) and (d) show the SEM images of MWA
with period of 1.3 μm and 3.3 μm, respectively. Page 14/18 Figure 2 Figure 2
Cut-off proton energy (a) and energy conversion efficiency (b) as a function of the thickness of planar SiN
foils (black) and the spatial period of MWA targets (red), from experiments (solid dots) and simulations
(solid lines). The error bars are defined by the level of uncertainty in different shots. The conversion
efficiency from laser to protons (kinetic energy >1 MeV) is obtained from the RCFs. Here the high
conversion efficiency for 0.2 μm results from more complicated mechanisms thus is excluded from
comparison. (c) Proton spectra obtained from RCF stacks (scattered points) and TP spectrometer
(dashed lines) for 1 μm SiN foils, and MWA targets with three periods of 1.3 μm, 2.3 μm, and 3.3 μm, Figure 2 Cut-off proton energy (a) and energy conversion efficiency (b) as a function of the thickness of planar SiN
foils (black) and the spatial period of MWA targets (red), from experiments (solid dots) and simulations
(solid lines). The error bars are defined by the level of uncertainty in different shots. The conversion
efficiency from laser to protons (kinetic energy >1 MeV) is obtained from the RCFs. Here the high
conversion efficiency for 0.2 μm results from more complicated mechanisms thus is excluded from
comparison. (c) Proton spectra obtained from RCF stacks (scattered points) and TP spectrometer
(dashed lines) for 1 μm SiN foils, and MWA targets with three periods of 1.3 μm, 2.3 μm, and 3.3 μm, Cut-off proton energy (a) and energy conversion efficiency (b) as a function of the thickness of planar Si
foils (black) and the spatial period of MWA targets (red), from experiments (solid dots) and simulations
(solid lines). The error bars are defined by the level of uncertainty in different shots. The conversion
efficiency from laser to protons (kinetic energy >1 MeV) is obtained from the RCFs. Here the high
conversion efficiency for 0.2 μm results from more complicated mechanisms thus is excluded from
comparison. (c) Proton spectra obtained from RCF stacks (scattered points) and TP spectrometer
(dashed lines) for 1 μm SiN foils, and MWA targets with three periods of 1.3 μm, 2.3 μm, and 3.3 μm, Page 15/18 Page 15/18 Page 15/18 respectively. The horizontal error bar marked for p=3.3 μm as the last sheet of RCF stack changed color in
this shot. (d) The simulated proton spectra corresponding to the conditions from (c). this shot. (d) The simulated proton spectra corresponding to the conditions from (c). Figure 3
Expanded pre-plasma of MWA targets after loading the pre-pulse in FLASH situation for p=1.3 μm (a) and
p=3.3 μm (b). Mass density distribution along y = 17 μm (black), 18 μm (blue) and 19 μm (red) for p=1.3
μm (c) and p=3.3 μm (d) corresponds with (a) and (b), respectively. The diffracted pre-pulse field Ex
within the MWA structure (c). The density distribution of pre-expanding MWA target based on the FLASH
results (d) which is inputted into PIC simulations The inset shows the pre plasma scale lengths where lh Figure 3 Figure 3 Expanded pre-plasma of MWA targets after loading the pre-pulse in FLASH situation for p=1.3 μm (a) and
p=3.3 μm (b). Mass density distribution along y = 17 μm (black), 18 μm (blue) and 19 μm (red) for p=1.3
μm (c) and p=3.3 μm (d) corresponds with (a) and (b), respectively. The diffracted pre-pulse field Ex
within the MWA structure (c). The density distribution of pre-expanding MWA target based on the FLASH
results (d), which is inputted into PIC simulations. The inset shows the pre-plasma scale lengths, where lh
and lx are for the pillars, and lf for the substrate flat. Page 16/18 Figure 4
(a) The temporal evolution of Ex corresponding to Fig. 5(a-c), which is averaged in an area of 2 μm (x)×8
μm (y) behind the substrate. (b) The electron spectra and temperature under pre-plasma conditions. (c)
Proton maximum energy as a function of lh and lx scale length. (d) Proton maximum energy versus wire
period with and without pre-plasma, while the diameter and height of the wire are 0.4 μm and 2.5 μm,
respectively. The conversion efficiencies from the simulations in (d) are calibrated with the average value
measured in experiment for p=3.3 μm which gives a fixed ratio for all simulation data. Figure 4 (a) The temporal evolution of Ex corresponding to Fig. 5(a-c), which is averaged in an area of 2 μm (x)×8
μm (y) behind the substrate. (b) The electron spectra and temperature under pre-plasma conditions. (c)
Proton maximum energy as a function of lh and lx scale length. (d) Proton maximum energy versus wire
period with and without pre-plasma, while the diameter and height of the wire are 0.4 μm and 2.5 μm,
respectively. The conversion efficiencies from the simulations in (d) are calibrated with the average value
measured in experiment for p=3.3 μm which gives a fixed ratio for all simulation data. Page 17/18 Figure 5
The distribution of longitudinal electric field Ex for 1 μm planar target (a) and p=3.3 μm MWA target with
pre-plasma (b) and without pre-plasma (c). The fields are normalized to the incident laser electric field
E0=4.1×1013 V/m. (d-f) Proton energy (MeV) distribution at t=300fs. Figure 5 The distribution of longitudinal electric field Ex for 1 μm planar target (a) and p=3.3 μm MWA target with
pre-plasma (b) and without pre-plasma (c). The fields are normalized to the incident laser electric field
E0=4.1×1013 V/m. (d-f) Proton energy (MeV) distribution at t=300fs. Page 18/18
|
W4283828479.txt
|
https://bcpublication.org/index.php/BM/article/download/1024/1027
|
en
|
Research hotspots and development trend of green marketing in China from 1993 to 2021: knowledge graph analysis based on Citespace
|
BCP business & management
| 2,022
|
cc-by
| 4,825
|
BCP Business & Management
Volume 20 (2022)
IEMSS 2022
Research hotspots and development trend of green marketing
in China from 1993 to 2021: knowledge graph analysis based
on Citespace
Niu Pei*
Nanjing University of Science and Technology, Nanjing, China
*Corresponding author: np_cumt@163.com
Abstract. As the whole world enters the era of environmental protection, the green revolution has
brought vitality to enterprises and society. Carrying out green marketing, producing green products
and setting up green marketing concepts are the hot topics of academic interest, and also a great
opportunity for enterprises. Using the visual analysis software citespace, draw the corresponding
knowledge map, carry out corresponding quantitative research on domestic green marketing
research, and analyze the knowledge structure and evolution pattern of green marketing research
in China from a bibliographic perspective. The results of the study show that there is little cooperation
between authors and institutions in the field of green marketing, . The research themes are mainly
distributed in green marketing, eco-marketing, low carbon economy, green consumption, green
products and other aspects, among which "green marketing" is the biggest node of continuous
attention. Future research should focus on the application and promotion of green marketing in
agricultural products, and at the same time, strengthen the connection and cooperation between
authors and institutions, broaden the breadth and depth of research, and study the development and
application of green marketing through more literature.
Keywords: Green marketing, Research hotspots, Development trend, Citespace knowledge graph,
Visual analytics.
1. Introduction
Originally, the word "green" emerged from the United States in the 1960s. Since then, interest in
its research has increased among economists, sociologists and marketing scholars, leading to its
development in the early 1980s (Leonidou & Leonidou, 2011). In the 1990s, green marketing became
a green movement, and the decade of 1990 was declared "the era of the Green Revolution"
(Vandermerwe & Oliff, 1990). Since then. Green marketing has become an important research field
for scholars (Fuller, 1999; Hartmann et al., 2005; Juwaheer et al., 2012; Peattie, 1995; Polonsky and
Mintu- Wimsatt, 1995). In November 1992, Chinese representatives attended the international
Marketing seminar held in Hong Kong, where domestic scholars first came into contact with the
concept of green marketing and carried out corresponding research [2]. In 1997, the first monograph
green Marketing was published, marking the formation of green marketing theory in China.
With the development and evolution of the research theory in this field, domestic researches on
green marketing have accumulated more achievements and formed a relatively mature system. By
combing through the relevant literature on green marketing research in China, it can be seen that the
research in this field is rich in content, diverse in research topics and involves multi-disciplinary cross
research. Although the existing research has a certain comprehensive analysis of domestic green
marketing [3], the relevant studies are mainly qualitative studies and lack of quantitative studies on
green marketing. Unable to capture the hot spots and trends of green marketing research.
Therefore, this research uses the Citespace information visualization software, and uses the SCI,
CSSCI, Peking University Core and CSCD included in China National Knowledge Infrastructure
(CNKI) as data sources to carry out a corresponding quantitative research on domestic green
marketing research, and draw a corresponding knowledge map. It displays the research hotspots,
research trends and evolution laws in this field with visual information, providing theoretical
reference for follow-up research and practical exploration.
497
BCP Business & Management
Volume 20 (2022)
IEMSS 2022
2. Definition of Green Marketing
Much of our understanding of green marketing comes from the perspective of consumers (Hartman
et al., 2005; Richeye et al., 2014). As a multifaceted concept, green marketing is linked to ecological
marketing, sustainable marketing, environmental strategies, environmental marketing and many other
terms (Abzari et al., 2013; Chamorroet al., 2009; Dangelico & Vocalelli, 2017; Garg, 2015). Peattie
(2010) divides green marketing into three stages: "ecological green marketing", "environmental green
marketing" and "sustainable green marketing". At present, the domestic academic circles have
basically reached a consensus on the definition of green marketing. They believe that a more
comprehensive definition is proposed by Wei Mingxia and Si Linsheng (2001): they believe that
green marketing means that under the requirements of the concept of sustainable development,
enterprises will take responsibility from the society. From the perspective of responsibility,
environmental protection, full use of resources, and long-term development, in the whole process of
product research, development, production, sales, and after-sales service, take corresponding
measures to achieve sustainable consumption of consumers, sustainable production of enterprises,
and complete The balance of three aspects of sustainable development of society. It aims to achieve
the effective allocation of limited resources and pursue the organic coordination of short-term
marketing behaviour and long-term marketing strategies with society, economy, resources and
environment as well as the benign influence on the long-term development of enterprises [3]. It can
be seen that green marketing is more focused on the coordination between the interests of consumers,
enterprises and the social environment. While meeting the needs of consumers, attention should be
paid to the ecologically balanced development of nature.
3. Data sources and research methods
3.1 Data sources and processing
The data sources of this study are mainly selected from journals with high quality literature in
China Knowledge Network (CNKI), including CSSCI, SCI, CSCD and Peking University Chinese
core journals. As Green Marketing is generally considered to be the Chinese translation of English
"Green Marketing". Some scholars also use "Environmental Marketing" (Environmental Marketing),
"lower-carbon Marketing" (low-carbon Marketing) and "Ecological Marketing" and other words to
express. Considering that green marketing, environmental marketing, low carbon marketing and ecomarketing are often blurred by scholars, these four keywords were searched simultaneously. The
search terms were limited to "green marketing" or "environmental marketing" or "eco-marketing" or
"low carbon marketing ". In order to make the data more credible, the data classification proposes
engineering technology, etc., and only retains the economic and management science literature.
Through precise matching, a total of 1300 literatures related to green marketing from 1993 to 2021
were retrieved, and the retrieval time was March 21, 2022. Through manual screening one by one,
irrelevant topics and no-author documents were eliminated, and finally 1040 valid documents were
screened.
3.2 Research method
CiteSpace is a visualization software developed by the famous Chinese American scholar Chaomei
Chen in Java language. It is mainly based on co-citation analysis theory and pathfinding network
algorithm, etc., to measure the literature (collection) of a specific field, in order to explore the key
path of the evolution of the discipline and its knowledge inflection point (represented by key papers),
and through a series of visualization mapping to form the analysis of the potential dynamics of the
discipline evolution and the detection of the frontier of the discipline development [5]. As a dedicated
scientific knowledge mapping tool, CiteSpace has been widely used by relevant research institutions
and personnel in the international scientometrics community since its launch in September 2004.
Therefore, with the help of CiteSpace software, this paper takes the core journals in the field of green
498
BCP Business & Management
Volume 20 (2022)
IEMSS 2022
marketing in China as the research object, and visualises the authors and institutions, research
hotspots and evolution patterns in this research field, so as to objectively show the current situation
and development trend of research in this field. The parameter selection (Selection Criteria) of
Citespace was Top50 per slice, and the running time was December 1, 2021, with a time span of
1993-2021 (Slice Length=1). The Pathfinder function was used for pruning and optimisation to
construct a co-occurrence map of authors, organisations and keywords in the field of green marketing,
and the corresponding clustering map was generated according to the keyword co-occurrence.
4. Analysis of research results
4.1 Analysis of the overall trend of domestic green marketing research
A corresponding analysis of the volume of domestic green marketing publications from 1993-2021,
according to the results of Figure 1, shows that the number of papers published increased year by year
from 1994 to 2001. The number of articles published in 2002 was a breaking point and a turning point,
after which the number of articles tended to first decrease and then increase, reaching a peak of 116
articles in 2007. After that, the number of articles published fluctuated, but the number of articles
published every year reached more than 50. But after 2011, the number of articles published reached
less than 20. However, the corresponding prediction of the trend of the number of articles shows that
the number of articles will show an increasing trend year by year after 2020. The data results are
shown in Figure 1:
Figure 1. Domestic green marketing publication volume, 1993-2021
4.2 Core authors and institutional analysis
(1) Visualization analysis of core authors
The citespace software was used to visualise the core authors, setting the Node types as Author
Author and selecting the top 50 frequency data in each time slice. Run the citespace software and get
the author distribution and cooperation map in the domestic green consumption field, as shown in
Figure 2. In the author distribution graph, the node represents the number of papers published by
authors in this field, and the larger the node, the more papers the authors publish; the line represents
the cooperation between authors, and the thicker the line, the stronger the cooperation between
authors. Conversely, the weaker the collaboration between the authors. By running citespace software,
an author distribution profile with 658 nodes, 204 connections and a network density of 0.0009 was
obtained.
The visualization map of core authors shows that the whole network distribution in the author
distribution map is loose, and there are many published authors, but not many highly productive
authors. By analysing and collating the literature, the top ten authors of the article were obtained,
499
BCP Business & Management
Volume 20 (2022)
IEMSS 2022
namely: Jing Shaoping, Deng Desheng, Li Shu, Wei Mingman, Si Linsheng, Rao Kefu, Shen Genrong,
Zhang Guohua, Zhang Juihuan and Yu Daisong. In the author distribution network map, the number
of connections between authors is also scattered, indicating that there is less cooperation between
authors in the field of green marketing, and a certain scale of cooperation has not yet formed. The
published literature is basically completed independently.
Figure 2. Distribution and co-occurrence mapping of green marketing research authors in China
(2) Visualization analysis of core institutions
In order to further analyze the institutional distribution and cooperation map in the domestic green
marketing research field, Set the Node types to Institution and leave all other parameters untouched.
Run Citespace and get the distribution and cooperation map of domestic green consumption
institutions, as shown in Figure 3. A distribution and cooperation map of institutions with 575 nodes,
154 connections and a network density of 0.0009 was obtained. In this research area, the top ten
institutions in terms of number of publications were School of Economics and Management of Harbin
Engineering University, the Department of Trade and Economics of Zhongnan University of
Economics and Law. Business School of Central South University, the School of Business of Central
South University of Forestry and Technology, the School of Finance and Economics of Anhui
University of Finance and Economics, the School of Economics and Finance of Xi'an Jiaotong
University, the School of Law of Shandong University of Technology, the School of Business
Administration of South China University of Technology, the School of Business of Jilin University
and the School of Economics and Management of Beijing Forestry University. The results of the
study show that there are many institutions issuing articles, but not many articles per institution, and
that the top ten institutions in terms of the number of articles issued are mostly colleges of university
management. In terms of institutional collaboration, institutions are spread out, mostly in a two-point
partnership, suggesting a lack of collaboration between institutions and less collaborative output.
500
BCP Business & Management
Volume 20 (2022)
IEMSS 2022
Figure 3. Distribution and cooperation network map of domestic green marketing research
institutions
5. Research Hotspots and Frontier Analysis
5.1 Research Hotspot Analysis
(1) Keywords analysis of domestic green marketing research
Select the top 50 frequency data in each time slice. Run Citespace to get the keyword distribution
map with 684 nodes, 1320 connections, and 0.0057 network density (as shown in Figure 4). Each
circular node represents a keyword and the frequency of the keyword is indicated by the size of the
circle, the larger the circle the more frequently the keyword appears and the more representative of
the research hotspots in the field of green marketing. The lines between the nodes indicate the cooccurrence (or co-introduction) relationship and their thickness indicates the strength of the cooccurrence (or co-introduction). The colour corresponds to the time of the first co-occurrence (or cointroduction) of the node. The change in colour from cool blue to warm red indicates a change in time
from early to recent.
Figure 4. Green Marketing Research Keyword Co-occurrence Map
501
BCP Business & Management
Volume 20 (2022)
IEMSS 2022
Figure 4 shows that the largest node in the mapping is green marketing, followed by green products,
green consumption, countermeasures, green barriers, consumers, companies, green management, and
marketing. The frequency of keyword appearance, that is, the size of the node, also reflects the
research hotspot in the field of green marketing to a certain extent. In addition, the keyword nodes in
the graph are purple circles indicating that the keyword has a high centrality. From Figure 4, it can
be concluded that the keywords with high centrality in the field of green marketing research are green
marketing, eco-marketing, low carbon economy, green consumption, green products,
countermeasures, green barriers, consumers, marketing strategies, innovation and other high
centrality keywords.
Comprehensive domestic green marketing research in the field of high-frequency keywords and
high-centricity keywords derived from the initial research in the field, ecological marketing, green
marketing, green products, green consumption, marketing and other mainstream research position,
and then promote marketing strategies, green management, ecological civilization, green barriers and
other research, and later with the study of countermeasures, innovation, enterprise and other research
to promote the research of low-carbon economy, supply chain and other research has become a green
marketing research field of research hotspots, and continue to promote the field of in-depth research.
(2) Hot spot cluster analysis of domestic green consumption research
Keyword clustering focuses on reflecting the structural characteristics and thematic distribution
among clusters, highlighting key nodes and important connections, and by doing corresponding
clustering analysis on keywords in the field of green marketing research to reveal the research topics
in this field. Run citespace software to obtain a keyword clustering distribution profile with 684 nodes,
1568 connections and a network density of 0.0067 (as shown in Figure 5). Q value is greater than 0.5,
S value is greater than 0.7, so the clustering is reasonable. Obtained 11 major cluster labels of
domestic green marketing research, including: green marketing, green barriers, green products,
network marketing, green consumption, low-carbon economy, marketing strategies, innovation,
green management, ecological marketing, and marketing.
Figure 5. Cluster map of domestic green marketing research hotspots
Analyze the research topics under each cluster tag, select the first five topics under each cluster
tag, as shown in Table 1. Based on further analysis of the corresponding literature, it is concluded
that green marketing research mainly focuses on traditional marketing, consumers, eco-tourism,
marketing and other research topics; Green barriers are mainly studied around countermeasures,
agricultural products, marketing and enterprises; green products are more concerned with consumer
demand and green marketing of enterprises; online marketing involves knowledge marketing,
relationship marketing, marketing innovation, knowledge economy and other research themes. Green
consumption involves more fields, including green marketing development countermeasures, tourism,
international marketing, etc. The research scope of low carbon economy covers supply chain, low
502
BCP Business & Management
Volume 20 (2022)
IEMSS 2022
carbon marketing, enterprise marketing and other topics; Marketing strategy pays more attention to
the formulation of strategy; Innovative research themes are mainly combined with the development
trend of the new century to determine the corresponding research themes, such as green
entrepreneurship, green management, etc.; green management includes green certification, strategies,
the current situation, hotels and other research themes; eco-marketing mainly focuses on the research
of products and product promotion, etc.; marketing involves various research themes, including
market economy, socialism, state-owned commercial banks, opportunities and other research themes.
Cluster
number
0
1
2
3
4
5
6
7
8
9
10
Table.1. Comparison of power load forecasting of 403 line
Cluster
Identifiers (select the first 5)
size
Green marketing, traditional marketing, consumers, ecotourism,
173
marketing
Green Barriers, Countermeasures, Agricultural Products, Marketing,
70
Business
Green products, consumers, green signs, green consumer demand,
58
corporate green marketing
Internet marketing, knowledge marketing, relationship marketing,
42
marketing innovation, knowledge economy
Green consumption, development strategies, international marketing,
37
promotion, tourism marketing
Low carbon economy, supply chain, evolutionary gaming, low carbon
37
marketing, corporate marketing
Marketing strategy, green logistics, brand marketing,
33
internationalization, localization
Innovation, ecological civilization, new century, green
31
entrepreneurship, green management
28
Green management, green certification, strategy, status, hotels
Eco-marketing, eco-products, green marketing, blockchain,
28
infomercials
Marketing, market economy, socialism, state-owned commercial
23
banks, opportunities
5.2 Research Frontier Analysis
(1)The sudden detection of domestic green marketing research keywords
Keyword burst detection can reflect the research fields with greater influence over a period of time.
A total of 15 emergent words (as shown in Figure 6.) are obtained through the corresponding keyword
emergent detection in the research field of green marketing. The top 10 keywords in terms of
prominence are: green barriers, low carbon economy, consumers, countermeasures, green products,
agricultural products, environmental protection, green labels, marketing strategies, and green logistics.
Judging from the time series of sudden keywords, before 1999, the sudden keywords were:
consumers, green signs, green products, environmental protection, green industry and knowledge
marketing; The emergent words from 2000 to 2010 are: knowledge economy, green management,
green barriers, countermeasures, green logistics; The sudden keywords in 2010 and after are: lowcarbon economy, marketing strategy, low-carbon marketing, agricultural products. According to the
keyword emergence degree and time node analysis, the domestic green marketing research field
mainly focuses on consumers, enterprises, green products and the development of corporate green
marketing strategies. Based on the time cycle of the sudden occurrence of keywords, agricultural
products emerge with the longest cycle (10 years), followed by green management, low carbon
economy and low carbon marketing with a cycle of 6 years, with most cycles concentrated around 4
503
BCP Business & Management
Volume 20 (2022)
IEMSS 2022
years. However, in general, green marketing research in China mainly focuses on consumers,
enterprises and environmental protection. In recent years, due to the national emphasis on green
economy and green products, agricultural products have also become a major research hotspot in the
field of green marketing research. For green and natural agricultural products, enterprises formulate
corresponding green marketing strategies to promote the healthy and balanced development of
enterprises, consumers and the environment, which is a hot topic in the field of green marketing
research.
Figure 6. 1994-2021 Top 15 emergent keywords
(3) The evolution of the frontier of domestic green marketing research and the division of research
stages
By analysing the evolutionary trends of research hotspots, that is, examining the temporal
distribution of high-frequency keywords and their trends, it is possible to grasp the research frontiers
and developments in the field. In this study, the keyword co-occurrence analysis of domestic green
marketing research literature generates Timezone mapping, as shown in Figures 7. Timezone
mapping is mainly a time zone mapping formed by gathering together the same time nodes and
arranging them in chronological order to determine the overall characteristics of the evolution of
green marketing research themes in China. Running citespace, we get a keyword co-occurrence time
zone map with Q values greater than 0.5 and S values greater than 0.7, so the clustering is reasonable.
The analysis in the diagram shows that the research hotspots in green marketing in China have
evolved over time as the research themes have developed and evolved. The Timezone diagram
provides a visual representation of the research hotspots within each time period. The largest node in
the graph is green marketing, which first appeared in 1994 and spans the longest time period and has
the largest flow to other research hotspots. Followed by hot research topics such as green products,
green consumption, countermeasures, and ecological marketing. Judging from the literature and
nodes etc. it is concluded that the domestic green marketing research process is mainly concerned
with the areas of enterprise, consumer, environment and green products. The development of green
marketing in China is broadly divided into three stages according to the hot keywords, emergent
words and the number of literature on green marketing in China in different time periods.
504
BCP Business & Management
Volume 20 (2022)
IEMSS 2022
Figure 7. 1994-2021 Domestic green marketing research keyword co-occurrence Timezone
mapping
Phase 1 (1993-1999): Green marketing has been widely recognised and studied since its
introduction in the 1980s and 1990s. A large number of high-frequency keywords in this research
area appear in this period, such as the appearance of high-frequency keywords such as green
marketing, green products, eco-marketing, green consumption and consumers. In terms of the volume
of literature, this period saw a modest but growing trend in the literature, laying an important
cornerstone for subsequent research in the field.
Phase II (2000-2010): This period was marked by the formulation of policies such as the 11th
Five-Year Plan and the need to change the mode of economic development and develop a green
economy. The number of green marketing studies in China has shown a rapid growth trend, and the
number of articles published has taken up a large proportion. High-frequency keywords such as green
management, agricultural products, green barriers, ecological civilisation and countermeasures
emerge. Began to focus on green marketing strategies and other research.
Phase 3 (2011-2021): In the context of the new economic normal, a green, low-carbon circular
economy has become a reality. After 2012, the ecological crisis caused by the high growth, high
energy consumption development model has intensified and generated new pressure, the haze formed
by the deterioration of the public environment has received global attention, the 18th National
Congress creatively put forward the concept of ecological civilization, which is the echo of the serious
situation in the environmental field, ecological construction was included in the central government's
governance of one of the five major layouts, becoming a basic state policy, China has truly entered
the era of low-carbon green growth.
6. Conclusion and outlook
6.1 Research conclusions
In this study, the core literature in the field of domestic green marketing research in China from
1994-2021 in CNKI was visualised and analysed accordingly by using bibliometric methods with the
help of citespace software. By mapping the corresponding networks, it visualises the research
hotspots and the evolution of research frontiers in the field of green marketing research in China. The
study found that:
In terms of research subjects, there are many authors working in the field of green marketing
research in China, but not many highly productive ones, and there is less cooperation between authors
505
BCP Business & Management
Volume 20 (2022)
IEMSS 2022
and a looser distribution. In terms of research institutions, the number of institutions publishing green
marketing research in China is relatively large, but most of them are independent, with little
cooperation between institutions, and most of the issuing institutions are university management
schools.
In terms of research hotspots, "green marketing" has always been a hot topic of concern for
domestic scholars, followed by green products, green consumption, countermeasures, green barriers,
consumers, enterprises and other topics closely related to green marketing. These themes are clustered
to form 11 categories: green marketing, green barriers, green products, online marketing, green
consumption, low carbon economy, marketing strategy, innovation, green management, ecomarketing, marketing, and each category contains the corresponding research theme. By doing a
sudden detection of keywords to reflect the more influential research areas, the results show that in
recent years, agricultural products have become a hot topic in the field of green research, the
application of green marketing strategies with agricultural products to promote green marketing
research areas. Through the analysis of the frontier evolution and development stage of domestic
green marketing, it is concluded that with the deduction and evolution of time, the research hotspots
in this field are also changing with time, and green marketing has always become a research hotspot
in academia. And according to the keyword co-occurrence Timezone mapping, green marketing
research is divided into three main development stages, each stage with the development of national
development policies and theoretical maturity derived from the corresponding research hotspots, thus
promoting the continuous progress of research in this field.
Through a visual analysis of the domestic green marketing research field, it can be concluded that
the research on green marketing in China is mainly focused on three levels, namely the consumer,
enterprise and environmental levels. At the consumer level, domestic studies have mainly focused on
consumers' mindset and perception of green consumption. At the enterprise level, existing literature
focuses on green marketing strategies, operational models, and strategies for enterprises to cope with
green barriers. At the environmental level, the main emphasis is on the balanced development of
enterprises and the environment, low-carbon and green development.
6.2 Shortcomings and outlook
By combing through the process of domestic research on green marketing, it can be concluded that
since 1993, domestic research on green marketing has become more and more in-depth, but there are
still corresponding research problems.
Since the 21st century, due to the concept of sustainable development and people's increasing
awareness of environmental protection, the market for green agricultural products in China has
continued to develop and grow, and has achieved some remarkable results in the development of
green marketing of agricultural products. However, compared to developed countries, China is still
at the initial stage of development of green marketing for agricultural products. The visual knowledge
map shows that agricultural products have become a hot research topic in the field of green marketing
research, and future research can be expanded accordingly for agricultural products, from the
perspective of enterprises, consumers and environmental protection. In addition, the mapping of
authors and institutions shows that there is less collaboration between authors and institutions. In the
future, academic collaboration and the building of academic teams should be further strengthened,
focusing on collaborative research among scholars from various institutions and different
backgrounds and emphasising multidisciplinary cross-sectional research.
The research in this paper also has corresponding shortcomings. The data sources in this paper are
mainly core journals published in China Knowledge Online, and due to the limitation of data format,
no citation analysis of the literature has been conducted to show the citation and cited relationship
between knowledge. In addition, this study did not include other types of literature, such as
dissertations and books, nor did it include research results published by Chinese scholars in
international journals; future studies can further compare and analyse through a wider range of
literature sources.
506
BCP Business & Management
Volume 20 (2022)
IEMSS 2022
References
[1] Zhang Bing, Lin Yihong. Reflections on the implementation of green marketing strategy in
Chinese enterprises [J]. Enterprise Economy, 2007 (06): 83-85.
[2] Li Mingliang. Research on green marketing in China: a review and outlook [J]. Journal of
Yunnan University of Finance and Economics, 2010, 26(01):125-131.
[3] Ji Shidi, Luo Yuanyuan, Li Xiaoyong. A review of green marketing research in China [J]. China
Business Journal, 2019 (06): 63-66.
[4] Wei Mingxia, Si Linsheng, Sun ShuSheng. Analysis of the basic categories of green marketing
[J]. Jiangxi Social Science, 2001(06): 88-90.
[5] Chen Yue, Chen Chaomei, Liu Zeyuan, Hu Zhigang, Wang Xianwen. Methodological functions
of CiteSpace knowledge graphs [J]. Scientology Research, 2015, 33(02):242-253.
[6] Richey Jr R G, Musgrove C F, Gillison S T, et al. The effects of environmental focus and program
timing on green marketing performance and the moderating role of resource commitment [J].
Industrial Marketing Management, 2014, 43(7): 1246-1257.
[7] Dangelico R M, Vocalelli D. “Green Marketing”: An analysis of definitions, strategy steps, and
tools through a systematic review of the literature [J]. Journal of Cleaner production, 2017, 165:
1263-1279.
[8] Hartmann P, Ibáñez V A, Sainz F J F. Green branding effects on attitude: functional versus
emotional positioning strategies [J]. Marketing intelligence & planning, 2005.
[9] Abzari M, Safari Shad F, Abedi Sharbiyani A A, et al. Studying the effect of green marketing
mix on market share increase [J]. European Online Journal of Natural and Social Sciences:
Proceedings, 2013, 2(3 (s)): pp. 641-653.
[10] Peattie K. Green consumption: behavior and norms [J]. Annual review of environment and
resources, 2010, 35: 195-228.
[11] Chamorro A, Rubio S, Miranda F J. Characteristics of research on green marketing [J]. Business
Strategy and the Environment, 2009, 18(4): 223-239.
[12] Garg, A. (2015). Green marketing for sustainable development: an industry perspective.
Sustainable Development, 23(5), 301-316.
507
|
|
https://openalex.org/W4394825977
|
https://openfuture.pubpub.org/pub/internet-standards/download/pdf
|
English
| null |
Finding the path to a more open internet. A new European approach toward internet standards
| null | 2,024
|
cc-by
| 10,718
|
Open Future
Finding the path to a more
open internet. A new
European approach toward
internet standards Open Future Please share feedback directly in the publication or by writing to Clément Perarnaud
(Clement.Perarnaud@vub.be). Please share feedback directly in the publication or by writing to Clément Perarnaud
(Clement.Perarnaud@vub.be). Cover illustration: Jakub Koźniewski / PanGenerator, CC BY Cover illustration: Jakub Koźniewski / PanGenerator, CC BY Abstract The purpose of this report is to frame a new vision for what could become the future of the European Union’s
(EU) Open Internet agenda, as part of the next European Commission’s mandate in 2024. Though also
acknowledging recent positive steps, this report challenges the consistency of the European approach toward
Open Internet standards and identifies a series of legislations and initiatives that seemingly contradict the
overall stance of the EU in favor of protecting the internet as a global, interoperable, and open network of
networks. The report identifies what could be the main building blocks for a renewed EU approach in view of
remediating those inconsistencies. It lists six specific areas in which the EU could support the development of
more “open” internet standards, referring both to the standards themselves and to the processes by which they
are formulated and adopted. These recommendations are expected to pave the way for an alternative path for
upcoming EU digital policies, conducive to more openness for the future of the internet. Acknowledgments: I thank Zuzanna Warso, Alicja Peszkowska, Paul Keller, and Alek Tarkowski for their
excellent feedback and support throughout the fellowship. I am also grateful to Julien Rossi for his always
insightful thoughts on the EU’s standardization approach and to the anonymous experts who shared their time
and comments on earlier drafts. 1Open Future Open Future Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Introduction The internet is often celebrated for its openness. Usually presented as one of the few critical properties having
propelled the internet’s success over recent decades, the notion of openness remains at the heart of its founding
mythology.1 Though for arguably different reasons, the openness of the internet is supported by a wide range of civil society
organizations, companies, and states, using this concept as a synonym for “free,” “interoperable,” or “market-
friendly.” Though for arguably different reasons, the openness of the internet is supported by a wide range of civil society
organizations, companies, and states, using this concept as a synonym for “free,” “interoperable,” or “market-
friendly.” Despite all of its supporters, there is an emerging consensus that the Open Internet is now under threat. Two
major developments are generally considered, namely the almost unparalleled power gained by a few
technological monopolies over the internet, and the acceleration of the assertiveness of state actors in relation
to its infrastructure and governance. In part justified by the former, the growing assertiveness of states is
evidenced by new claims supporting the protection of their digital sovereignty. Though there are of course
different understandings of what it means in practice, the concept of digital sovereignty generally refers to the 2 Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Open Future ambition that states and governments should reaffirm their authority and protect their self-determination in the
digital sphere.2 Such claims are generally voiced as an alternative to a “hegemonic” status quo,3 at times
identified as the ever-growing power of Big Tech, and/or the broader dominance of powerful states such as the
U.S. or China over the internet. ambition that states and governments should reaffirm their authority and protect their self-determination in the
digital sphere.2 Such claims are generally voiced as an alternative to a “hegemonic” status quo,3 at times
identified as the ever-growing power of Big Tech, and/or the broader dominance of powerful states such as the
U.S. or China over the internet. While India, Russia, and China have been actively developing their own sovereign internet infrastructures4
over recent years, the EU has also become a strong advocate of its own declination of digital sovereignty, and
notably since the beginning of the Commission’s von der Leyen presidency in 2019. Introduction Departing from its traditional market-driven approach applied to digital policies,5 this shift has led to forceful
attempts to curb the power of large technological companies over European digital markets, but also to
increase the EU’s influence over global internet standard-setting processes, as will be investigated throughout
this report. Standards are indeed at the center of many states’ and companies’ strategies to exert global influence and shape
the internet “in their own image.”6 Standards allow for the interoperability between the tens of thousands of
autonomous systems connecting worldwide. Standards have become arguably the most important pieces of the
infrastructure making the internet a global network of networks. Due to their economic and strategic
importance, internet standards are thus inevitably the subject and reflection of intense economic and political
battles. This report studies the implications of having an EU policy agenda characterized by new sovereignty claims in
this domain and explores the specific case of internet standardization. Identifying clear inconsistencies between
the EU’s aspiration to promote Open Internet standards and its most recent digital policy trajectory, the report
proposes a series of avenues to develop a renewed EU vision conducive to more openness for the internet. The remainder of the report is structured as follows. The next section provides an analysis of the EU’s
approach toward internet standards in the context of its recent policy agenda centered since 2019 around the
notion of digital sovereignty. After challenging the trajectory and consistency of its policies, the report
identifies what could be the main building blocks for a renewed EU approach fostering more openness in the
field of internet standardization. Unpacking the EU’s approach toward internet standards: when
digital sovereignty meets the Open Internet. In her State of the Union Address in 2020, the European Commission president Ursula von der Leyen
emphatically voiced the EU’s objective to “secure [its] digital sovereignty” and “lead the way on digital – or it
will have to follow the way of others, who are setting these standards for us.” Those words illustrate the clear
process of “geopoliticization” of EU digital policies7 that could be observed under her presidency, bringing
digital issues in the remits of geopolitical competition. 3 Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Open Future In 2022, the European Commissioner Thierry Breton illustrated this new geopolitical European approach to
standardization by stating that: “In French we say ‘qui fait la norme, détient le marché’: ‘who makes the
standard holds the market.’ […] If we want to ensure Europe's technological sovereignty in crucial disruptive
sectors such as 5G, batteries, hydrogen or quantum technology, we must occupy the field of standard-setting. We must become standard-makers, and not just standard-takers.” This discourse, followed by many others, also underlined that the rationale for the EU approach has been
primarily defensive (“defend the open, decentralized internet”) and framed as a reaction to two main
developments. First, EU discourses on internet standards usually factor proposals voiced by Chinese entities to
transform core protocols on which the internet relies, initiatives that are part of a broader impetus from China
to gain power through international standardization. But if China is now considered as an emerging security
and strategic challenge for the EU, as highlighted by the adoption of the EU toolbox for 5G security in 2020, it
is not the only justification for a stronger EU policy agenda in this area. The EU approach also identifies the
dominance of large U.S.-based technological companies on digital markets as a clear policy issue. It justified
the adoption of a brand new repertoire of legislative measures, such as the 2022 Digital Markets Act (DMA),
as well as other lesser-known initiatives with equally important impacts on internet standards, detailed below. Thus, the policy narrative of the European Commission clearly emphasized the need for the EU to set global
standards in order to achieve its own digital sovereignty. Unpacking the EU’s approach toward internet standards: when
digital sovereignty meets the Open Internet. Though it is difficult to characterize the EU approach
to digital sovereignty, it is usually qualified8 as the EU’s attempt to regain control over the digital field and
develop international leadership capacity. In this context, an over-reliance on standards set outside of the
European continent is presented as clashing with the protection of a global, open, free, and decentralized
internet, in stark contrast with other state actors such as Russia and China, treating digital sovereignty as
equivalent to territorial sovereignty9 over the internet. This discursive connection between standards and digital sovereignty in the European policy agenda has
materialized in a number of areas and through different instruments. The following section presents the most
relevant EU initiatives adopted between 2019 and 2023.10 It also discusses the coherence and consistency of
the overall EU approach in relation to the protection of the internet’s openness. EU standardization strategy and digital policies Over recent years, the EU has been particularly active in the digital policy space, with direct implications for
internet standards. This is primarily illustrated by the 2022 European standardization strategy but also by other
digital policy legislations such as the Digital Markets Act and the eIDAS regulation. In contrast with its predecessor, the 2022 EU standardization strategy explicitly identifies internet
standardization as a key area of interest. It states that “a particularly critical situation relates to internet
standardization to promote a free, open, accessible, inclusive and secure global Internet” and warns about the 4 Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Open Future increasing politicization of “the international standardisation [of] Internet protocols,” which could limit “the
evolution of the global open Internet.” increasing politicization of “the international standardisation [of] Internet protocols,” which could limit “the
evolution of the global open Internet.” One of the pillars of the strategy focuses on coordination between EU institutions and European Member
States, emphasizing the need to “strengthen the EU’s voice in global standardisation.” This part is reflective of
the fact that currently, “EU Member States allocate varying amounts of resources to the international
standardisation bodies, and the European Union as such does not have a formal voice in multilateral fora.” This applies both to international standard-developing organizations (SDOs), such as the Internet Engineering
Task Force (IETF), as well as European standardization organizations (ESOs), as underlined in the next
paragraph. As the only organizations eligible to work on standardization requests from the European Commission, we
know that the European Telecommunications Standards Institute (ETSI), the European Committee for
Standardization (CEN), and the European Committee for Electrotechnical Standardization (CENELEC) are
increasingly visible, as a result of their important role in the implementation phase of recent and upcoming EU
digital legislations (such as the AI Act). Due to their significant importance, the 2022 standardization strategy
indicates the intention of the European executive to restructure the decision-making process of these European
SDOs, and, in particular, ETSI. Indeed, the Commission had recently expressed concerns that ETSI’s decision-
making system granted a disproportionate voting power to “certain corporate interests.” Huawei is, for
instance, a founding member of ETSI’s Industry Specification Group on Securing Artificial Intelligence. EU standardization strategy and digital policies While these recent attempts by the Commission could create momentum in support of a form of regionalization
of global standardization, the inherent contradictions of the EU approach underline, however, the limited
capacity of the EU to isolate itself from global standardization processes. First, recent changes in ETSI’s
leadership are not really indicative of a new trajectory, as a representative from the U.S. company Intel has
become ETSI’s chair in December 2023. In addition, it should be noted that many European standards –
including harmonized standards – directly refer to specifications from global, and arguably U.S.-led, consortia,
underlying the great reliance of the European standardization system on global private SDOs to develop state-
of-the-art specifications. While these recent attempts by the Commission could create momentum in support of a form of regionalization
of global standardization, the inherent contradictions of the EU approach underline, however, the limited
capacity of the EU to isolate itself from global standardization processes. First, recent changes in ETSI’s
leadership are not really indicative of a new trajectory, as a representative from the U.S. company Intel has
become ETSI’s chair in December 2023. In addition, it should be noted that many European standards –
including harmonized standards – directly refer to specifications from global, and arguably U.S.-led, consortia,
underlying the great reliance of the European standardization system on global private SDOs to develop state-
of-the-art specifications. Aside from the standardization strategy, the EU has adopted a few important legislations with direct
implications for internet standards and internet SDOs. The first is the 2022 Digital Markets Act (DMA), one of
the flagship initiatives of the EU to curb the monopoly powers of Big Tech actors (labeled as “gatekeepers”) in
European digital markets. In this context, the DMA requires interoperability between message services from
the most used messaging apps, such as Meta’s Messenger or WhatsApp. Though not formally requested as
such by the European Commission, the actual standardization of an interoperable protocol has been initiated by
a few actors of this industry in the IETF in 2022 as part of the MIMI working group. This example illustrates a
little-known channel through which the EU digital sovereignty agenda is directly influencing the internet
standardization field. It is an important case of an EU sovereignty-oriented legislation directly affecting the
work of the IETF, a traditionally industry-driven organization with a global reach. EU standardization strategy and digital policies In another important EU legislation, the eIDAS regulation, discussions about EU-approved certificate
authorities for web browsers have been on the policy agenda. This followed a proposal by the European
Commission to force web browsers to approve government-approved qualified website authentication
certificates (QWACs), rather than preserving their freedom as to which certificates they could recognize as
secure to protect communications. Justified by cybersecurity goals, this initiative has been challenged12 by
both the technical community and civil society organizations due to the political implications of this change for
privacy, notably because it could allow third parties to intercept and modify traffic without the knowledge or
consent of the sender and recipient.13 Though the compromise adopted in trilogues in November 2023
eventually states that “the requirement to QWACs does not affect browser security policies,” it signals another
clear attempt by the EU to alter internet standards on the grounds of protecting its digital sovereignty. EU standardization strategy and digital policies Despite its ESO status, the Commission argued that ETSI’s governance and membership cannot be understood
as strictly European. Based on this approach, the recent adoption of a targeted amendment to Regulation
1025/2012 on European standardization stated that for an ESO to be eligible for standardzation requests from
the Commission, its internal governance must ensure that the national standardization bodies of European
countries will “hold the decision-making power in each stage of the development of a standard requested.”11
This means the decision-making process of ETSI, which generally consisted of giving one vote per company
(regardless of where it is headquartered), would have to change when dealing with standards requested by the
Commission. This evolution challenges, to a certain extent, the scope and raison d’être of ETSI, which had
projected itself over the years as a global SDO, rather than an European one. This particular focus on ETSI could suggest that the influence of foreign companies in CEN-CENELEC is less
of a challenge. Yet, non-EU companies are often active directly at the level of national standardization bodies,
and their views thus get to be represented regularly at the European level through CEN-CENELEC, though via
more indirect means. The EU standardization strategy may have direct implications for the process of making internet standards, as
ETSI has historically been a major actor in the field of mobile internet standards, but also at the transport layer 5 Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Open Future of the internet in the development of new encryption protocols. From this standpoint, the strategy seems to
conflict with the broader policy discourse of the EU regarding the global and open internet, as it seemingly
pushes in favor of a regionalization of technological standardization that could affect internet infrastructures. of the internet in the development of new encryption protocols. From this standpoint, the strategy seems to
conflict with the broader policy discourse of the EU regarding the global and open internet, as it seemingly
pushes in favor of a regionalization of technological standardization that could affect internet infrastructures. EU research and innovation projects Besides legislations, the EU approach in relation to internet standards has also taken the form of a series of
research and innovation projects. Some can be framed as additional concretizations of the EU digital 6 Finding the path to a more open internet. A new European
approach toward internet standards Open Future sovereignty agenda. sovereignty agenda. The European Commission has, for instance, identified a set of specific internet standards, understood as the
right vehicles for protecting the Open Internet through its new EU Internet Standards Deployment Monitoring
Website. This monitoring website provides deployment indicators of 18 key internet standards that will “help
to secure the Internet and support its constant technological evolution.” This tool is purely descriptive, but
shows the priority given by the Commission toward five categories of standards: 1. Browsing – Web
communication standards, 2. Routing – Mutually Agreed Norms for Routing Security (MANRS), 3. Emailing
– Email communication security standards, 4. Naming – Domain Name System Security Extensions
(DNSSEC), and 5. Addressing – Internet Protocol version 6 (IPv6). In parallel, StandICT, a recent EU-funded project launched in 2018, has aimed to increase European experts'
presence in international standardization in the broad field of ICT. It addresses the fact that funding can be an
important barrier to participation for individuals and small companies (along with technical expertise or
language). This project is now considering internet standardization as a priority, in view to add more European
voices to internet SDOs. Up until now, this funding stream had rarely benefited IETF participants, but there is
now an intention to address this issue. Finally, another important EU-funded project is the DNS4EU initiative. It is not strictly focused on internet
standardization as such, but still refers to a number of key objectives related to internet standards. Featured in
the 2020 EU Cybersecurity Strategy for the Digital Decade, this initiative has the ambition to develop a public
European Domain Name System (DNS) resolver service. The function of a DNS resolver is to “resolve” the
queries of users when they connect to a website, by translating domain names into IP addresses. The DNS
resolver market is known for its rapid concentration, leading a handful of companies – based mostly outside of
the EU – to control a significant portion of the internet traffic. EU research and innovation projects Though the initial intention to deconsolidate the market of DNS resolvers is laudable, this project reveals a
number of inconsistencies in the EU approach. Though it has been introduced on the grounds of increasing
cybersecurity and privacy protection, as well as avoiding an over-reliance on the solutions provided by U.S.-
based companies (Google in the first place), it could nonetheless create a worrying precedent. Indeed, while it
intends to contribute to the normalization of privacy-protecting protocols developed by the internet community
(including the IETF), it is at the same time providing pathways and normalizing discourses for state actors to
directly intervene in the internet architecture.14Despite of its potential implications, it should be acknowledged
that the actual realisation of this new DNS resolver appears to be, to this day, at a standstill. International cooperation and technological diplomacy Aside from these research and innovation projects, the EU has been active in relation to internet standards
through a myriad of initiatives and collaborations carried out with third countries. The 2021 EU Global
Gateway initiative – which mirrors the approach of the Chinese Belt and Road Initiative (BRI) and its Digital 7 Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Open Future Silk Road – is one of them. The EU Global Gateway is expected15 to link investments into technological
infrastructure with the deployment of standards and rules in third countries. The Commission has explicitly
stated that investments in digital infrastructure will be “linked with standards and protocols that support
network security and resilience, interoperability, and an open, plural and secure Internet.” Though the actual
concretizations of this initiative remain limited so far in the area of digital infrastructure, this initiative shows
that, conceptually, the deployment of certain internet standards is now being advanced through digital
infrastructure investments in developing countries (now framed as “partner countries” in EU jargon). The launch of the EU-US Trade and Technology Council (TTC) in 2021 also suggests evolutions in the EU
approach. Though its mandate is broader, this transatlantic mechanism is expected to promote the alignment of
EU-U.S. positions within international standard-setting bodies, including internet SDOs, and for instance
through the new Strategic Standards Information mechanism. Once again, the actual outcome of this new
cooperation stream remains to be seen, but it nonetheless echoes the growing geopolitical nature of internet
standards for the EU. Finally, the European Commission, through DG CONNECT and DG INTPA, has also initiated a series of
lobbying and public relation campaigns toward third countries focusing on internet standards. The EU is
attempting to engage with specific countries,16 mostly on the African continent, in view of upcoming ITU
plenaries – the prime objective being to counter the growing influence of China in this fora. The recent European Commission’s international approach toward internet standards is thus much more
geopolitical than in the past. At the same time, attempts to enhance the EU’s digital sovereignty create clear
frictions with the European vision favoring the promotion of an Open Internet. International cooperation and technological diplomacy If defending the internet’s openness was and remains a common denominator at the European level, this report
points to a series of inconsistencies, some of which are highlighted in the following table, questioning the
coherence of the EU’s policy approach. 8
Table 1: Overview of the recent EU approach in relation to internet standards. Type
Initiative
Key impacts in relation to internet
standards Table 1: Overview of the recent EU approach in relation to internet standards. Type 8 Finding the path to a more open internet. A new European
approach toward internet standards Open Future This diverse set of initiatives highlights the way in which the EU policy agenda has been shaped by new
sovereignty claims in this domain, with admittedly varying levels of ambition and success. It also appears clear
EU legislations and policy strategy
2022 standardization strategy
2022 Digital Markets Act (DMA)
eIDAS regulation
push in favor of a regionalization of
technological standardization, that
could affect internet infrastructures
introduce interoperability provision
directly influencing the work of the
IETF
proposal of EU-approved certificate
authorities for web browsers could
allow third parties to intercept and
modify traffic
EU research and innovation projects
Internet Standards Deployment
Monitoring
StandICT
DNS4EU
provide descriptive deployment
indicators of 18 key internet standards
support increase of European experts'
presence in international
standardization
create a new public DNS resolver, but
also provide pathways and normalize
discourses for state actors to directly
intervene in the internet architecture
International tech diplomacy and global
cooperation
Global Gateway
EU-US TTC
ITU diplomacy
support the deployment of certain
internet standards through digital
infrastructure investments in
developing countries
promote the alignment of EU-U.S. positions within international
standard-setting bodies, including
internet SDOs
counter the growing influence of
China in the ITU through influence
campaigns proposal of EU-approved certificate
authorities for web browsers could
allow third parties to intercept and
modify traffic support increase of European experts'
presence in international
standardization create a new public DNS resolver, but
also provide pathways and normalize
discourses for state actors to directly
intervene in the internet architecture support the deployment of certain
internet standards through digital
infrastructure investments in
developing countries promote the alignment of EU-U.S. Building blocks for a new European approach toward open internet
standards Building blocks for a new European approach toward open internet
standards It is commonly admitted that the policy approach of the EU in relation to the digital economy, and more
broadly to the internet, can have a great impact worldwide. The much-commented on “Brussels Effect”
popularized by Anu Bradford17 is a testimony to the fact that EU norms can have a significant influence well
beyond European borders, in part due to the still significant economic power of its market. With its new agenda promoting digital sovereignty, we have shown how the EU is normalizing a set of
discourses and technical interventions related to the internet that can be construed as a challenge to its
openness. Though in different ways, this is illustrated by recent initiatives and projects such as the DNS4EU,
the eIDAS, or the EU standardization strategy. Having identified areas in which the EU policy approach generates inconsistencies, we now reflect on avenues
for the next European Commission to reimagine its approach in the near future. The report identifies six areas
in which the EU could support the development of more open internet standards, referring both to the standards
themselves and to the processes by which they are formulated and adopted. The first part reflects on what
should be the role – and even responsibility – of the EU in promoting certain types of internet standards. The
second focuses on how to better reflect the public interest in private standardization processes. International cooperation and technological diplomacy positions within international
standard-setting bodies, including
internet SDOs counter the growing influence of
China in the ITU through influence
campaigns This diverse set of initiatives highlights the way in which the EU policy agenda has been shaped by new
sovereignty claims in this domain, with admittedly varying levels of ambition and success. It also appears clear
that, while certain attempts provide for useful policy gestures, others may be seen as incompatible with the
broader public policy message of the EU in relation to the open and global internet. This is especially
problematic as today’s internet is already known to be characterized by a series of patterns that gradually limit
its openness. They include the consolidation of its infrastructures in the hands of a small set of companies, the 9 Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Open Future various claims for more network control expressed by states, and the privatization of decision-making in
relation to global internet standards. In response to these challenges and inconsistencies, the following section lays down the building blocks of
what could be a new European vision conducive to more openness in relation to the internet. Supporting standards conducive to more internet openness As exemplified by the 2022 EU standardization strategy or the EU Global Gateway initiative, the Commission
has recently developed a series of channels by which it aims to support the deployment of certain standards
across the EU and abroad. In this context, specific internet standards are being supported by the European
Commission at technical and political levels. It is currently unclear what the criteria are on which the current selection of standards is based, besides
fostering the growth of the network and its security. Though these objectives are entirely legitimate, this report
argues that the EU would gain from prioritizing a broader set of policy objectives, and thus a more diverse
range of standards. This should be done through the various ways in which the EU can promote the
deployment of internet standards, considering for instance public procurements and investments in digital
infrastructure. 10 10 Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Open Future As fostering interconnectivity should not be an end for itself, one would need to make sure that the internet
standards supported by the EU are aligned with the overarching goal of the development of an open, people-
centered internet. EU-supported internet standards need to have broader objectives, including to foster the de-
consolidation of the internet, push for greater cooperativity, and support, unequivocally, end users’
privacy. These objectives could be operationalized through the various means currently at the disposal of the
EU, including public procurements, and as part of a dedicated strategy on internet standardization. b. Moving away from the interconnection narrative While internet SDOs are formidable machines to create consensus on how to make the internet grow and
“work better” (the official goal of the IETF), they appear much less equipped in relation to other types of
societal or political norms. Discussions within and outside of the human rights protocol considerations (HRPC)
research group of the Internet Research Task Force (IRTF) are a case in point of the relative resistance from
many corporate engineers to see their work framed as political24 and their opposition to the notion that specific
political values can be enshrined in internet standards. Yet, we know that internet standards can constitute substantive political issues25 with direct implications for
state actors, companies, or individuals alike. In this context, drawing on its human-centric inclination with
respect to its digital policies, the EU should support the development and deployment of internet standards that
are not solely focused on connecting more devices and making the network grow. This could be
operationalized, for instance, through dedicated impact assessments. As convincingly argued by Paris et al.,26
instead of being only fixated on the constant growth of global interconnection, standards should be oriented
toward supporting more cooperativity between individuals and local communities, for example. Internet standards have become in many ways “the default infrastructure for society.”27 Limiting latency or
ensuring the security of communications are evidently positive objectives, and arguably in line with the
interests of most internet users. Yet, the – rather understandable – fixation of internet SDOs on the efficiency
and growth of the network prevents discussions on the actual implications of their deployment. The
anthropologist Corinne Cath argues28 in this sense that internet SDOs, such as the IETF, have engineered their
own innocence in relation to the actual use and societal implications of the standards they formulate. The EU approach should recognize this challenge by thinking more in terms of qualitative connectivity rather
than plain quantitative interconnection, being more mindful of the magnitude of both positive and negative
impacts generated by the internet’s expansion across the world. As presented below, this shift would also
require thinking of the internet not only as a critical infrastructure to be made bigger and more efficient but also
recognize it as a space where individuals are increasingly subject to various forms of control and surveillance. a. De-consolidating the internet If it was to convert its internal digital competition policy into action on the global standardization front, the EU
could support the formulation and deployment of standards explicitly aiming for the de-consolidation of the
internet. Internet consolidation refers to a process of increasing control over internet infrastructure and
services18 by a small set of organizations. The internet is known to be increasingly consolidated, or in other
words, moving toward a larger fraction of traffic involving a smaller set of large content providers, social
networks, and content delivery platforms. In the IETF, and within other industry-driven internet SDOs, it is common to see large technological
companies pushing for their own standards to become the norm for the rest of the internet.19 There is a relative
consensus20 about the fact that only a few technological companies effectively have the capabilities to develop
sophisticated technical proposals and push them in internet SDOs. This situation is further amplified by the quasi-total control that large corporate actors leverage over certain
parts of the internet, as a result of the growing consolidation of its many markets and infrastructure (such as
DNS resolvers services as seen above). Thus, supporting standards that directly contribute to the consolidation of the internet should not be on the
European agenda. This does not mean that all standards promoted by large companies have to be considered
negatively by default, especially given the current structure of the private internet governance regime,21 which
structurally empowers large technological companies to be responsible for most internet standards. Even in this constrained realm, the focus of the EU should be to support standards that are aimed precisely at
the de-consolidation of the internet. These efforts should be cognizant of the fact that “even when open
protocols incorporate techniques intended to prevent consolidation, economic and social factors can drive users
to prefer solutions built with or on top of supposedly decentralised technology.”22 Yet, and as indicated
previously, the example provided by the EU Digital Markets Act is an inspiring example of how the crucial
aspiration for interoperability23 can be politically prescribed by public regulators and become addressed as a
result by internet SDOs to address worrying patterns of consolidation. 11 11 Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Open Future c. More encryption, less network control Standards supported by the EU need to empower individuals and end users who are confronted with the
challenges of an increasingly digitized world. If internet SDOs, such as the IETF, have relatively failed to
integrate societal norms and values into their agenda, they have been at the forefront of discussions on
encryption and security. In the wake of the Snowden revelations a decade ago, internet SDOs have developed
sophisticated ways to encrypt and protect communications – which have become standard practices across the
world and which the European Commission has also supported over recent years by promoting IETF’s
DNSSEC specifications to secure DNS data, for instance. 12 Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Open Future Yet, in the meantime, EU Member States and specific Directorates-General (DGs) of the European
Commission (including the DG for Migration and Home Affairs) have promoted a policy agenda that appears
adversarial to the deployment of encryption techniques over the internet, while favoring more network control. This fight against encryption is front and center in the current EU negotiations on the CSAM regulation, for
example. Instead of weakening encryption, requesting backdoors, or considering the use of secure messaging
tools (such as Signal) as a self-incriminating act (see in France), the EU should vocally support people-
centered privacy-protecting protocols. This indirectly relates to discussions occurring in more formal and state-centered internet SDOs, such as the
Technical committee of the International Telecommunication Union (ITU-T). New technical proposals led by
China29 (labeled as “New IP”) underline the ambition of a number of countries to reshape fundamental
protocols on which the internet relies to get more visibility and control over the networks. Countering these
developments has been presented by the EU as an important political priority. The European Commission has
invested significant political capital at the international level to leverage influence over these technical
processes and prevent proposals that favor more state-based network control from becoming a reality. This
report highlights that the direction of the EU’s internal policies in relation to encryption weakens its
simultaneous claims grounded in human rights against state-centered network control in the context of internet
SDOs such as the ITU. c. More encryption, less network control This section thus argues that adding these three new dimensions – de-consolidation of the internet, push for
greater cooperativity, and support of individuals’ privacy and autonomy – would bring added value and greater
coherence to the EU agenda in relation to internet standards. But what also matters is how these standards are adopted and the “openness” of these processes for different
types of stakeholders – and, most importantly, civil society – in contributing to their formulation. a. De-privatizing standard-setting The literature on the feudalization of the internet30 describes at length the power and control accumulated by
large technological companies over the internet, which evolved, according to Tim Wu,31 from “a freely
accessible channel to one strictly controlled by a single corporation or cartel – from open to closed system.” The approach of the EU in relation to technology standardization has been grounded for decades, if not longer,
in the belief that industries are better suited to lead such processes. Micklitz makes the argument32 that “the
relocation of knowledge from the administration to industry is the result of a process that dates back to the late
19th century and begins with industrialization,” underlining the emergence of vast discrepancies in the
technical expertise held within state bureaucracies and large technological companies.33 In relation to the internet, the current legitimacy of this largely private-driven standardization process falls
short. The words of Ben Tarnoff34 and his critique of the industry-dominated internet are essential in this
context: “Understanding how privatization made the modern internet is essential for any movement that seeks
to remake it. Movements must know their enemy. If they expend their energy on the wrong target, the
opportunity for meaningful change is lost. History shows why privatization is the right target, how it forms the
common foundation for the diverse dysfunctions and depredations of the modern internet.” Though a strictly multilateral governance regime for internet standards is evidently not a desirable option from
a human rights standpoint, the EU should support the emergence of an internet standardization regime that is
not captured – or even saturated – by a few multinational corporations. This effort could take the form of
enhanced political exchanges with internet SDOs to discuss how their policies related to inclusion and
transparency could be improved, based on the practices and guidelines developed by the EU through its
European standardization system, for instance. These exchanges could be nurtured through bilateral activities,
similar to those of the Internet Society, aimed at familiarizing policymakers with the work of the IETF, and
vice-versa. This recommendation does not suggest, however, that the general practices of ESOs in relation to participation
and transparency are to be systematically followed by internet SDOs. The openness of the standardization
processes of both ETSI and CEN-CENELEC is notoriously problematic. Neither the minutes of working group
meetings nor their actual composition are made public. Supporting more openness in internet standardization processes In the field of internet standardization, the current status quo – namely the predominant private industry-driven
regime – falls short in terms of democratic accountability, legitimacy, and inclusion. As evidenced by the 2022 standardization strategy and the role given to ESOs in the context of new EU digital
policies, participation and representativeness in technological standardization processes are an important area
of priority for the European Commission. However, as for other domains, the EU’s approach appears
characterized by several inconsistencies resulting from competing political, security, and economic objectives
presented above. The new European Commission approach toward internet standards should support the
opening of new ways to effectively contribute to the making of internet standards for actors that are not large
technological corporations. These efforts should aim at de-privatizing standard-setting, broadening
participation, and politicizing standards. 13 Finding the path to a more open internet. A new European
approach toward internet standards Open Future b. Broadening participation While there are merits to reaffirming the EU’s agency in standard-setting, the EU discourse often fails to
recognize and address the unintended consequences of its own initiatives, such as accelerating the growing
“trend in state-based rule-setting on Internet infrastructure.”35 Instead of normalizing the power of digital
monopolies or state-based approaches to internet infrastructure, the EU should support civil society and citizen
engagement and their effective participation in internet standardization processes. While the project StandICT is a first step in the direction of supporting more European voices (with
presumably limited means) to attend these standardization arenas, the limited scale of this project, which funds
a hundred experts for the whole ICT standardization field, underscores the limits of the current EU approach. Mirroring the comments made by Micklitz36 in 2023 in reference to EU technical standardization policy,
current initiatives designed to secure the meaningful participation of all concerned stakeholders in internet
SDOs looks more “like a fig leaf” than a structural policy. For instance, the input of European civil organizations within IETF is infinitesimal, and greater funding should
be allocated in order to foster the effective participation of civil society and citizens in this area. Also, the EU should not remain strictly focused on the European scale when it comes to participation and
funding support. The EU would benefit from broadening participation in standardization processes to actors
that are usually not included in these technical arenas, regardless of their location, as long as they promote an
internet centered around people and the public interest. a. De-privatizing standard-setting The standards themselves are often protected by
intellectual property rights and are not made available free of charge. Admittedly, this is in stark contrast with
the practices of other SDOs such as the IETF or W3C. Nonetheless, EU policies in relation to the inclusion of
non-commercial stakeholders, granting them special rights and formal membership in the standard
development process, could be an interesting avenue to consider for such SDOs. 14 14 Finding the path to a more open internet. A new European
approach toward internet standards Open Future c. Politicizing standards Internet SDOs, such as the IETF, are not used to answer the normative, political, or societal issues that their
standards may generate – a pattern that also characterizes technological standardization as a whole. Internet
SDOs, like international organisations,37 often depoliticize their work as a way to keep themselves out of
politics. This is reflected in their organizational cultures and the professional habits of their individual
members. This has positive implications – for instance, in terms of efficiency and trust – that fall short in
comparison to the negative aspects, including the dire lack of accountability of these processes. The EU should support the politicization of these technical discussions. Politicizing does not mean making
them subject to the direct control of political governments or state actors. On the contrary, it means formulating
them in the open and raising the public awareness of the political implications of those technical deliberations,
allowing individuals and social groups to be aware and discuss their implications, and support relevant
communities to participate in these arenas. Participation from all the various concerned publics seems essential
before such standards become effectively enacted and gradually shape the invisible infrastructure of the
internet. 15 15 Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Open Future The work of Jean-Christophe Graz and Christophe Hauert has shown how the “importance of socio-
technological choices enacted in standards gives civil society organizations a strong incentive to be involved”
in standard-setting. Despite obvious resource asymmetries, it is common practice to see “reputation-sensitive
firms” accommodating (at the margins) some civil society demands in standardization processes, partly as a
way to legitimize those processes and give them greater credibility. These successes could fuel more
participation from non-commercial actors. Yet, despite these obvious benefits, civil society participation does not have to be imposed at any cost. Indeed,
the risk run by civil society actors to become instrumentalized in industry-driven standardization processes is
real. The participation of civil society actors needs to be supported in contexts where their contribution will be
valued and recognized, and where their positive contribution may trump their indirect support and legitimation
of such private governance arrangements. c. Politicizing standards The fact that almost no European civil society organizations can be found in the list of IETF participants over
recent years is a testimony of the necessary steps to be taken to inform public interest stakeholders that would
clearly benefit from being exposed to these technical conversations. The EU could improve its policies by
communicating (in different languages) about the political issues currently at stake in internet SDOs, or
directly support projects that contribute to the simplification of technical discussions, as exemplified by Article
19’s new Internet Standards Almanac. But awareness is evidently not the only obstacle preventing the
participation of such groups. The lack of technical expertise – and the often-missing involvement of public
interest technologists – within civil society organizations is a major challenge in Europe and beyond. In the
context of a shrinking space for civil society and worries about the lack of funding currently available for
digital rights organizations, the EU should address this challenge through more direct funding and capacity-
building activities directed toward civil society. These six high-level policy recommendations show that the EU would greatly benefit from a renewed policy
approach toward internet standards, with the aim of strengthening the internet’s openness, as further elaborated
in the concluding section. IV. Conclusion: Building an alternative imaginary and trajectory for
the future of the open internet “Making it possible for the world’s computers to talk to one another was an impressive technical achievement. Making this machinic conversation serve an end other than infinite accumulation will be a political one. It may
seem unlikely, but so was the internet. History is filled with improbable turns that look inevitable in retrospect. The future will be too.” (Tarnoff, 2022) The internet is contingent. It is the result of technological and political choices and imaginaries that, over the
course of half a century, gradually shaped its unique structure and the affordances it grants to its multitude of 16 Finding the path to a more open internet. A new European
approach toward internet standards Open Future users – be they states, companies, or individuals. users – be they states, companies, or individuals. The current status quo for internet standard-setting, namely the predominant private industry-driven regime,
falls short in terms of democratic accountability, legitimacy, and inclusion. This model also does not appear
well-equipped to address worrying patterns of market consolidation and power shifts favoring a few large
technological companies. Over recent years, the EU has entered this policy field in various ways, and at times precisely to address these
well-identified challenges. In doing so, this report underlines that the EU has recently attempted to assert itself
into the internet architecture, with both positive and negative implications, while pushing its agenda for EU
digital sovereignty. These attempts have created inconsistencies, and even incompatibilities, between the EU’s
policy objectives of asserting more control through and over the internet, and its overarching goal to protect it
as a global and open network. These inconsistencies highlight the fact that the EU is still torn between different objectives – which limits the
consistency and success of its policies. While a global supporter of multistakeholder internet governance, the
EU is pushing toward a form of regionalization of standard-setting through the 2022 EU standardization
strategy. While promoting the deployment of state-of-the-art encryption protocols, the Commission proposes to
weaken the use of encryption through the recent CSAM legislative proposal. While flexing its regulatory
muscles to curb the power of Big Tech, it does not structurally challenge its predominance in internet standard-
setting but rather normalizes the outcomes of existing industry-driven processes. IV. Conclusion: Building an alternative imaginary and trajectory for
the future of the open internet In 2024, the next European Commission’s mandate will give an important opportunity for the EU to frame a
new vision for what could become the future of the European Open Internet agenda. This report provides
recommendations that are expected to pave the way for an alternative path for EU digital policies, conducive to
more openness for the future of the internet. Table 1 summarizes what should be the main avenues of this new
approach. It contributes to the emerging body of work, supported in part by the Open Future Foundation,
aiming at re-imagining the internet and building infrastructures for the public good. Table 1: Six avenues for a new European approach toward internet standards Table 1: Six avenues for a new European approach toward internet standards
1. Supporting standards fostering more internet openness
De-consolidating the internet
Support the deployment of standards explicitly aiming to
address internet consolidation, and prescribe interoperability
when relevant. Moving away from the interconnection narrative
Support “meaningful connectivity” rather than plain
quantitative interconnection to foster cooperativity through the
networks. 1. Supporting standards fostering more internet openness Support the deployment of standards explicitly aiming to
address internet consolidation, and prescribe interoperability
when relevant. Support “meaningful connectivity” rather than plain
quantitative interconnection to foster cooperativity through the
networks. 17 Finding the path to a more open internet. A new European
approach toward internet standards Open Future Finding the path to a more open internet. A new European
approach toward internet standards Support people-centered privacy-protecting protocols instead
of promoting an adversarial policy approach to the deployment
of encryption techniques. More encryption, less network control Create exchanges with private internet SDOs to discuss how
inclusion and transparency could be improved – for instance,
based on the practices and guidelines developed by the EU. De-privatizing standard-setting Allocate greater funding to foster the effective participation of
civil society and citizens in this area, both at European and
global levels. Improve EU policies to communicate around the political
issues in internet SDOs, and support projects that contribute to
raising the awareness around these technical discussions. Bibliography: Abbate, Janet. Inventing the Internet. MIT Press, 2000. Abbate, Janet. Inventing the Internet. MIT Press, 2000. Andersdotter, Amelia, and Lukasz Olejnik. "Policy strategies for value-based technology standards." Internet
Policy Review 10, no. 3 (2021): 1-26. Arkko, Jari. “The Influence of Internet Architecture on Centralised versus Distributed Internet
Services.” Journal of Cyber Policy 5, no. 1 (2 January 2020): 30–45. Barrinha, André, and George Christou. "Speaking sovereignty: the EU in the cyber domain." European Security
31, no. 3 (2022): 356-376. Bartley, Tim. “Power and the Practice of Transnational Private Regulation.” New Political Economy 27, no. 2
(2022): 188-202. Bellanova, Rocco, Helena Carrapico, and Denis Duez. "Digital/sovereignty and European security integration:
an introduction." European Security 31, no. 3 (2022): 337-355. Bradford, Anu. The Brussels Effect: How the European Union Rules the World. Oxford University Press, 2020. Broeders, Dennis. The public core of the Internet: An international agenda for Internet governance. Amsterdam University Press, 2016. 18 18 Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Open Future Broeders, Dennis, Fabio Cristiano, and Monica Kaminska. "In Search of Digital Sovereignty and Strategic
Autonomy: Normative Power Europe to the Test of Its Geopolitical Ambitions." JCMS: Journal of Common
Market Studies 61, no. 5 (2023): 1261-80. Cath, Corinne. Changing Minds and Machines. Doctoral dissertation, University of Oxford, 2021. Cath, Corinne. Eaten by the Internet. Meatspace Press, 2023. Christou, George, and Seamus Simpson. "Gaining a stake in global internet governance: the EU, ICANN and
strategic norm manipulation." European Journal of Communication 22, no. 2 (2007): 147-164. Claessen, Eva. "Reshaping the internet–the impact of the securitisation of internet infrastructure on approaches
to internet governance: the case of Russia and the EU." Journal of Cyber Policy 5, no. 1 (2020): 140-157. Couture, Stephane, and Sophie Toupin. "What does the notion of ‘sovereignty’ mean when referring to the
digital?" New Media & Society 21, no. 10 (2019): 2305-2322. Daucé, Françoise, and Francesca Musiani. "Infrastructure-embedded control, circumvention and sovereignty in
the Russian Internet: An introduction." First Monday 26, no. 5 (2021). Doctorow, Cory. The Internet Con: How to Seize the Means of Computation. Verso Books, 2023. Graz, Jean-Christophe & Christophe Hauert. “Translating Technical Diplomacy: The Participation of Civil
Society Organisations in International Standardisation.” Global Society 33, no. 2 (2019): 163-183. Haggart, Blayne, Natasha Tusikov, and Jan Aart Scholte, eds. Bibliography: Power and Authority in Internet Governance:
Return of the State? 1st ed. Routledge, 2021. Harcourt, Alison, George Christou, and Seamus Simpson. Global Standard Setting in Internet Governance. Oxford University Press, 2020. Hoffmann, Stacie, Dominique Lazanski, and Emily Taylor. "Standardising the splinternet: how China’s
technical standards could fragment the internet." Journal of Cyber Policy 5, no. 2 (2020): 239-264. Karjalainen, Tyyne. "European Norms Trap? EU Connectivity Policies and the Case of the Global Gateway."
East Asia (2023): 1-24. Mager, Astrid. “European Search? How to counter-imagine and counteract hegemonic search with European
search engine projects.” Big Data & Society 10, no. 1 (2023). Micklitz, Hans-W. The Role of Standards in Future EU Digital Policy Legislation. Report of ANEC and
BEUC, 2023. URL: https://www.beuc.eu/sites/default/files/publications/BEUC-X-2023-
096_The_Role_of_Standards_in_Future_EU_Digital_Policy_Legislation.pdf 19 Finding the path to a more open internet. A new European
approach toward internet standards Open Future Musiani, Francesca. "Infrastructuring digital sovereignty: a research agenda for an infrastructure-based
sociology of digital self-determination practices." Information, Communication & Society 25, no. 6 (2022):
785-800. Musiani, Francesca. "Infrastructuring digital sovereignty: a research agenda for an infrastructure-based
sociology of digital self-determination practices." Information, Communication & Society 25, no. 6 (2022):
785-800. Nanni, Riccardo. "Digital sovereignty and Internet standards: normative implications of public-private relations
among Chinese stakeholders in the Internet Engineering Task Force." Information, Communication & Society
25, no. 16 (2022): 2342-2362. Nanni, Riccardo. "Digital sovereignty and Internet standards: normative implications of public-private relations
among Chinese stakeholders in the Internet Engineering Task Force." Information, Communication & Society
25, no. 16 (2022): 2342-2362. Paris, Britt S., Corinne Cath, and Sarah Myers West. "Radical infrastructure: Building beyond the failures of
past imaginaries for networked communication." New Media & Society (2023). Perarnaud, Clément, and Julien Rossi. "The EU and Internet standards–Beyond the spin, a strategic turn?"
Journal of European Public Policy (2023): 1-25. Perarnaud, Clément, Julien Rossi, Francesca Musiani, and Lucien Castex. "'Splinternets': Addressing the
renewed debate on internet fragmentation." Parlement Européen; Panel for the Future of Science and
Technology (STOA), 2022. DOI: https://data.europa.eu/doi/10.2861/183513 Russell, Andrew L. Open Standards and the Digital Age. Cambridge University Press, 2014. Seidl, Timo, and Luuk Schmitz. "Moving on to not fall behind? Technological sovereignty and the ‘geo-
dirigiste’ turn in EU industrial policy." Journal of European Public Policy (2023): 1-28. Tarnoff, Ben. Internet for the People: The Fight for Our Digital Future. Verso Books, 2022. ten Oever, Niels. Bibliography: Wired Norms: Inscription, resistance, and subversion in the governance of the Internet
infrastructure. Doctoral dissertation, University of Amsterdam, 2020. ten Oever, Niels. "Norm conflict in the governance of transnational and distributed infrastructures: the case of
Internet routing." Globalizations 20, no. 1 (2023): 184-200. Tréguer, Félix. L'utopie déchue. Fayard, 2019. Wu, Tim. The Master Switch: The Rise and Fall of Information Empires. Vintage, 2011. Wu, Tim. The Master Switch: The Rise and Fall of Information Empires. Vintage, 2011. Footnotes 1. Russell, Andrew L. Open Standards and the Digital Age. Cambridge University Press, 2014. ↩
2. Musiani, Francesca. "Infrastructuring digital sovereignty: a research agenda for an infrastructure-based
sociology of digital self-determination practices." Information, Communication & Society 25, no. 6 (2022):
785-800. ↩ 1. Russell, Andrew L. Open Standards and the Digital Age. Cambridge University Press, 2014. ↩
2. Musiani, Francesca. "Infrastructuring digital sovereignty: a research agenda for an infrastructure-based
sociology of digital self-determination practices." Information, Communication & Society 25, no. 6 (2022):
785-800. ↩ 20 Finding the path to a more open internet. A new European
approach toward internet standards Finding the path to a more open internet. A new European
approach toward internet standards Open Future 3. Couture, Stephane, and Sophie Toupin. "What does the notion of ‘sovereignty’ mean when referring to
the digital?" New Media & Society 21, no. 10 (2019): 2305-2322. ↩ 3. Couture, Stephane, and Sophie Toupin. "What does the notion of ‘sovereignty’ mean when referring to
the digital?" New Media & Society 21, no. 10 (2019): 2305-2322. ↩ 4. Nanni, Riccardo. "Digital sovereignty and Internet standards: normative implications of public-private
relations among Chinese stakeholders in the Internet Engineering Task Force." Information, Communication
& Society 25, no. 16 (2022): 2342-2362. ↩ 5. Seidl, Timo, and Luuk Schmitz. "Moving on to not fall behind? Technological sovereignty and the ‘geo-
dirigiste’ turn in EU industrial policy." Journal of European Public Policy (2023): 1-28. ↩
6. Broeders, Dennis. The public core of the internet: An international agenda for internet governance. Amsterdam University Press, 2016. ↩ 7. Broeders, Dennis, Fabio Cristiano, and Monica Kaminska. "In Search of Digital Sovereignty and
Strategic Autonomy: Normative Power Europe to the Test of Its Geopolitical Ambitions." JCMS: Journal of
Common Market Studies 61, no. 5 (2023): 1261–80. ↩ 8. Bellanova, Rocco, Helena Carrapico, and Denis Duez. "Digital/sovereignty and European security
integration: an introduction." European Security 31, no. 3 (2022): 337-355. ↩
9 ibid 9. ibid. ↩ 9. ibid. ↩ 9. b d. 10 This section partly draws on a recent academic publication co authored with Dr Julien Rossi 10. This section partly draws on a recent academic publication, co-authored with Dr. Julien Rossi, 0. This section partly draws on a recent academic publication, co-authored with Dr. Julien Ross investigating the interplay between the EU and internet standards. ↩ investigating the interplay between the EU and internet standards. ↩ 11. Footnotes The Commission expects this measure to grant more power to European actors in ESOs, and particularly
within ETSI, where the national delegation principle does not apply. ↩ 12. Internet Society. “Internet Impact Brief: Mandated Browser Root Certificates in the European Union’s
eIDAS Regulation on the Internet.”. ↩ 13. Other important legislations, such as the Cyber Resilience Act and the Artificial Intelligence Act, also
have a number of implications for technological standard-setting, but they remain out of the scope of this
report, which is focused on internet standards per se. For more see: https://www.ceps.eu/with-the-ai-act-we-
need-to-mind-the-standards-gap/ ↩ 14. Perarnaud, Clément, and Julien Rossi. "The EU and Internet standards–Beyond the spin, a strategic
turn?" Journal of European Public Policy (2023): 1-25. ↩ 15. Karjalainen, Tyyne. "European Norms Trap? EU Connectivity Policies and the Case of the Global
Gateway." East Asia (2023): 1-24. ↩ 21 21 Finding the path to a more open internet. A new European
approach toward internet standards Open Future 16. Perarnaud, Clement, and Julien Rossi. "The EU and Internet standards–Beyond the spin, a strategic
turn?" Journal of European Public Policy (2023): 1-25. ↩ 16. Perarnaud, Clement, and Julien Rossi. "The EU and Internet standards–Beyond the spin, a strategic
turn?" Journal of European Public Policy (2023): 1-25. ↩ 17. Bradford, Anu. The Brussels Effect: How the European Union Rules the World. Oxford University
Press, 2020. ↩ 17. Bradford, Anu. The Brussels Effect: How the European Union Rules the World. Oxford University
Press, 2020. ↩ 18. Arkko, Jari. “The Influence of Internet Architecture on Centralised versus Distributed Internet
Services.” Journal of Cyber Policy 5, no. 1 (2 January 2020): 30–45. ↩ 18. Arkko, Jari. “The Influence of Internet Architecture on Centralised versus Distributed Internet
Services.” Journal of Cyber Policy 5, no. 1 (2 January 2020): 30–45. ↩ 19. Harcourt, Alison, George Christou, and Seamus Simpson. Global Standard Setting in Internet
Governance. Oxford University Press, 2020. ↩ 19. Harcourt, Alison, George Christou, and Seamus Simpson. Global Standard Setting in Internet
Governance. Oxford University Press, 2020. ↩ 20. Cath, Corinne. Eaten by the Internet. Meatspace Press, 2023. ↩ 21. Haggart, Blayne, Natasha Tusikov, and Jan Aart Scholte, eds. Power and Authority in Internet
Governance: Return of the State? 1st ed. Routledge, 2021. ↩ 21. Haggart, Blayne, Natasha Tusikov, and Jan Aart Scholte, eds. Power and Authority in Internet
Governance: Return of the State? 1st ed. Routledge, 2021. ↩ 22. Nottingham, Mark. 1. Wu, Tim. The Master Switch: The Rise and Fall of Information Empires. Vintage, 2011. ↩ Footnotes “Internet Consolidation: What Can Standards Efforts Do?” Internet Draft. Internet
Engineering Task Force, 4 December 2022. https://datatracker.ietf.org/doc/draft-nottingham-avoiding-
internet-centralization/06/. ↩ 22. Nottingham, Mark. “Internet Consolidation: What Can Standards Efforts Do?” Internet Draft. Internet
Engineering Task Force, 4 December 2022. https://datatracker.ietf.org/doc/draft-nottingham-avoiding-
internet-centralization/06/. ↩ 23. Doctorow, Cory. The Internet Con: How to Seize the Means of Computation. Verso Books, 2023. ↩
24. Cath, Corinne. Changing Minds and Machines. Doctoral dissertation, University of Oxford, 2021. ↩
25. DeNardis, Laura. Protocol Politics: The Globalization of Internet Governance. MIT Press, 2009. ↩
26. Paris, Britt S., Corinne Cath, and Sarah Myers West. "Radical infrastructure: Building beyond the
failures of past imaginaries for networked communication." New Media & Society (2023). ↩
27 C h C
i
“E
b
h I
P
i
I
I f
P
Y
R d
” T
h P li 23. Doctorow, Cory. The Internet Con: How to Seize the Means of Computation. Verso Books, 2023. ↩
24. Cath, Corinne. Changing Minds and Machines. Doctoral dissertation, University of Oxford, 2021. ↩
25. DeNardis, Laura. Protocol Politics: The Globalization of Internet Governance. MIT Press, 2009. ↩
26. Paris, Britt S., Corinne Cath, and Sarah Myers West. "Radical infrastructure: Building beyond the
failures of past imaginaries for networked communication." New Media & Society (2023). ↩
27. Cath, Corinne. “Eaten by the Internet: Putting Internet Infrastructure Power on Your Radar.” Tech Policy
Press, 30 October 2023. https://techpolicy.press/eaten-by-the-internet-putting-internet-infrastructure-power-
on-your-radar. ↩ 28. Cath, Corinne. Changing Minds and Machines. Doctoral dissertation, University of Oxford, 2021. ↩
29. Nanni, Riccardo. "Digital sovereignty and Internet standards: normative implications of public-private
relations among Chinese stakeholders in the Internet Engineering Task Force." Information, Communication
& Society 25, no. 16 (2022): 2342-2362. ↩ 28. Cath, Corinne. Changing Minds and Machines. Doctoral dissertation, University of Oxford, 2021. ↩
29. Nanni, Riccardo. "Digital sovereignty and Internet standards: normative implications of public-private
relations among Chinese stakeholders in the Internet Engineering Task Force." Information, Communication
& Society 25, no. 16 (2022): 2342-2362. ↩ 30. Tréguer, Félix. L’utopie Déchue. Fayard, 2019. ↩ 30. Tréguer, Félix. L’utopie Déchue. Fayard, 2019. ↩ 31. Wu, Tim. The Master Switch: The Rise and Fall of Information Empires. Vintage, 2011. ↩ 22 Finding the path to a more open internet. A new European
approach toward internet standards Open Future 32. Micklitz, Hans-W. The Role of Standards in Future EU Digital Policy Legislation. Report of ANEC and
BEUC, 2023. ↩ 32. Micklitz, Hans-W. Footnotes The Role of Standards in Future EU Digital Policy Legislation. Report of ANEC and
BEUC, 2023. ↩ 32. Micklitz, Hans-W. The Role of Standards in Future EU Digital Policy Legislation. Report of ANEC and
BEUC, 2023. ↩ 33. A similar argument is made by Mariana Mazzucato in The Entrepreneurial State. ↩ 33. A similar argument is made by Mariana Mazzucato in The Entrepreneurial State. ↩ 34. Tarnoff, Ben. Internet for the People: The Fight for Our Digital Future. Verso Books, 2022. ↩ 34. Tarnoff, Ben. Internet for the People: The Fight for Our Digital Future. Verso Books, 2022. ↩ 35. ten Oever, Niels. Wired Norms: Inscription, resistance, and subversion in the governance of the Internet
infrastructure. Doctoral dissertation, University of Amsterdam, 2020. ↩ 35. ten Oever, Niels. Wired Norms: Inscription, resistance, and subversion in the governance of the Internet
infrastructure. Doctoral dissertation, University of Amsterdam, 2020. ↩ 36. Micklitz, Hans-W. The Role of Standards in Future EU Digital Policy Legislation. Report of ANEC and
BEUC, 2023. ↩ 36. Micklitz, Hans-W. The Role of Standards in Future EU Digital Policy Legislation. Report of ANEC and
BEUC, 2023. ↩ 37. Louis, Marieke, and Lucile Maertens. Why International Organizations Hate Politics: Depoliticizing the
World. Routledge, Taylor & Francis Group, 2021. ↩ 37. Louis, Marieke, and Lucile Maertens. Why International Organizations Hate Politics: Depoliticizing the
World. Routledge, Taylor & Francis Group, 2021. ↩ 23 23
|
https://openalex.org/W2000312857
|
https://escholarship.org/content/qt07c0q891/qt07c0q891.pdf?t=nu5mak
|
English
| null |
Dependence of century-scale projections of the Greenland ice sheet on its thermal regime
|
Journal of Glaciology/Journal of glaciology
| 2,013
|
cc-by
| 8,969
|
UC Irvine UC Irvine Previously Published Works Title
Dependence of century-scale projections of the Greenland ice sheet on its thermal
regime
li k Copyright Information
This work is made available under the terms of a Creative Commons Attribution License,
availalbe at https://creativecommons.org/licenses/by/4.0/ Peer reviewed Powered by the California Digital Library
University of California UC Irvine
UC Irvine Previously Published Works
Title
Dependence of century-scale projections of the Greenland ice sheet on its thermal
regime
Permalink
https://escholarship.org/uc/item/07c0q891
Journal
Journal of Glaciology, 59(218)
ISSN
0022-1430
Authors
Seroussi, H
Morlighem, M
Rignot, E
et al.
Publication Date
2013
DOI
10.3189/2013jog13j054
Copyright Information
This work is made available under the terms of a Creative Commons Attribution Lic
availalbe at https://creativecommons.org/licenses/by/4.0/
Peer reviewed UC Irvine
UC Irvine Previously Published Works
Title
Dependence of century-scale projections of the Greenland ice sheet on its thermal
regime
Permalink
https://escholarship.org/uc/item/07c0q891
Journal
Journal of Glaciology, 59(218)
ISSN
0022-1430
Authors
Seroussi, H
Morlighem, M
Rignot, E
et al. Publication Date
2013
DOI
10.3189/2013jog13j054
Copyright Information
This work is made available under the terms of a Creative Commons Attribution Lic
availalbe at https://creativecommons.org/licenses/by/4.0/
Peer reviewed UC Irvine
UC Irvine Previously Published Works
Title
Dependence of century-scale projections of the Green
regime
Permalink
https://escholarship.org/uc/item/07c0q891
Journal
Journal of Glaciology, 59(218)
ISSN
0022-1430
Authors
Seroussi, H
Morlighem, M
Rignot, E
et al. Publication Date
2013
DOI
10.3189/2013jog13j054
Copyright Information
This work is made available under the terms of a Crea
availalbe at https://creativecommons.org/licenses/by/
Peer reviewed INTRODUCTION varies between 5 and 40 km, due to intrinsic limitations of
SIA and computational requirements. Previous studies (Ritz
and others, 2001; DeConto and Pollard, 2003; Pollard and
DeConto, 2009; Applegate and others, 2012; Rogozhina and
others, 2012) illustrated the ability of these simplified models
to improve our understanding of ice-sheet paleoclimate, as
well as to evaluate the effect of different model parameters. Recent observations show an accelerated ice loss of the
Greenland ice sheet (Velicogna, 2009; Rignot and others,
2011) during the past few decades, attributed to both
changes in surface mass balance and ice dynamics (Howat
and others, 2007; Rignot and others, 2011). Retreat of outlet
glaciers is observed all around the periphery of the Green-
land ice sheet; however, the acceleration has a high
temporal and spatial variability (McFadden and others,
2011; Moon and others, 2012). Potential triggering mechan-
isms include, among others, increased calving rate (Benn
and others, 2007), intrusion of warm water in the fjords
(Holland and others, 2008), enhanced basal lubrication
(Schoof, 2010) and cryo-hydrologic warming of ice (Phillips
and others, 2010). One major shortcoming of these models is that they
operate at a relatively coarse spatial resolution compared
with the size of outlet glaciers that control the ice discharge,
which affect the total ice-sheet mass balance. Moreover,
these models fail to reproduce observed dynamical changes
of Greenland outlet glaciers where the model assumptions
break down and where glacier size is comparable to the
model spatial resolution (Gillet-Chaulet and others, 2012). In order to better simulate the ice dynamics, several next-
generation numerical models have been developed (Price
and others, 2011; Gillet-Chaulet and others, 2012; Larour
and others, 2012a; Seddik and others, 2012). These models
include improved three-dimensional (3-D) models (e.g. higher-order (HO; Blatter, 1995; Pattyn, 2003) and full-
Stokes (FS) models), and use anisotropic mesh refinement
(Morlighem and others, 2010) to capture narrow outlet
glaciers at the ice-sheet periphery. Contrary to SIA- or SSA-
based models, HO and FS models are too computationally
intensive to perform interglacial spin-ups. In order to perform
realistic simulations and start from a suitable representation
of present-day conditions, these models either rely on spin-
ups of simpler models (Seddik and others, 2012) or use data
assimilation to reproduce present-day conditions (Morligh-
em and others, 2010; Larour and others, 2012a). H. SEROUSSI,1 M. MORLIGHEM,2 E. RIGNOT,2;1 A. KHAZENDAR,1 E. LAROUR,1
J. MOUGINOT2 1Jet Propulsion Laboratory, California Institute of Technology, Pasadena, CA, USA
E-mail: helene.seroussi@jpl.nasa.gov
2Department of Earth System Science, University of California, Irvine, Irvine, CA, USA ABSTRACT. Observations show that the Greenland ice sheet has been losing mass at an increasing rate
over the past few decades, which makes it a major contributor to sea-level rise. Here we use a three-
dimensional higher-order ice-flow model, adaptive mesh refinement and inverse methods to accurately
reproduce the present-day ice flow of the Greenland ice sheet. We investigate the effect of the ice
thermal regime on (1) basal sliding inversion and (2) projections over the next 100 years. We show that
steady-state temperatures based on present-day conditions allow a reasonable representation of the
thermal regime and that both basal conditions and century-scale projections are weakly sensitive to
small changes in the initial temperature field, compared with changes in atmospheric conditions or
basal sliding. We conclude that although more englacial temperature measurements should be acquired
to validate the models, and a better estimation of geothermal heat flux is needed, it is reasonable to use
steady-state temperature profiles for short-term projections, as external forcings remain the main
drivers of the changes occurring in Greenland. Powered by the California Digital Library
University of California eScholarship.org eScholarship.org 1024 Journal of Glaciology, Vol. 59, No. 218, 2013
doi: 10.3189/2013JoG13J054 INTRODUCTION The first
method captures past changes of the ice sheet, but leads to
discrepancies between the thermal regime and the stress
regime. Indeed, the spin-up model and the model that is used
for future scenarios have different spatial resolutions and rely
on different ice-flow approximations, leading to an initial To better understand the behavior of the Greenland ice
sheet, several large-scale ice-flow numerical models have
been employed to simulate the evolution of the ice sheet over
the coming centuries (Greve, 1997a; Ritz and others, 2001;
Greve and others, 2011; Gillet-Chaulet and others, 2012;
Seddik and others, 2012). These models rely on different ice-
flow approximations, parameterizations of physical pro-
cesses and initialization procedures. Most ice-flow models
rely on simplified first-order approximations of the mo-
mentum balance equations, such as the shallow-ice approx-
imation (SIA; Hutter, 1982), the shelfy-stream approximation
(SSA; MacAyeal, 1989; Greve and others, 2011; Aschwan-
den and others, 2012a) or a combination of SIA and SSA
(Bueler and Brown, 2009; Pollard and DeConto, 2009). Initialization of these numerical models generally consists of
running paleoclimate spin-ups over at least the last glacial
cycle, in order to obtain a suitable present-day configuration
(Greve and others, 2011; Aschwanden and others, 2012a). With this method, long-term memory of the ice-sheet
evolution is included in several model variables (e.g. ice
temperature). The spatial resolution of these models usually 1025 Seroussi and others: Greenland thermal regime for future projections horizontal and vertical velocities are decoupled. The ice
viscosity, , is assumed to be isotropic and to follow Glen’s
flow law (Glen, 1955): shock that influences the results (Seddik and others, 2012). The stress regime, and therefore the viscous and basal
heating, is not consistent with the initial temperature. The
second method consists of using present-day geometry of the
ice sheet (ice thickness, surface elevation, ice front position)
and inferring unknown parameters (e.g. basal friction) to
reproduce observed velocities with data assimilation (Mac-
Ayeal, 1993a; Morlighem and others, 2010). As this method
relies solely on present-day conditions, past climate vari-
ability is not accounted for. Temperatures are recovered using
a thermal steady state (Morlighem and others, 2010; Larour
and others, 2012a), even though the ice is not in thermal
equilibrium. INTRODUCTION The ice velocity and geometry computed using
such a method are consistent with present-day observations;
however, inconsistencies between velocity and thickness
datasets lead to ice flux divergence anomalies and models
start by artificially redistributing the glacier mass, not as a
realistic projection but to reconcile the inconsistencies
(Seroussi and others, 2011). ¼
B
2_"
n 1
n
e
±,
ð1Þ ð1Þ where
_"e is the effective strain rate, n is Glen’s law
coefficient (taken as n ¼ 3) and B is the ice hardness. B is
mainly temperature-dependent and we rely on the relation-
ship provided by Cuffey and Paterson (2010, p. 75). The
horizontal velocity is a solution of r 2_"1
ð
Þ ¼ g @s
@x ,
r 2_"2
ð
Þ ¼ g @s
@y ,
ð2Þ ð2Þ with with _"1 ¼
2@u
@x þ @v
@y
1
2
@u
@y þ @v
@x
@u
@z
2
66666664
3
77777775
_"2 ¼
1
2
@u
@y þ @v
@x
@u
@x þ 2@v
@y
@v
@z
2
66666664
3
77777775
,
ð3Þ The ice-sheet thermal regime has a large effect on the ice
flow, as it affects basal melting and therefore the sliding of ice
over its bed, and it also controls ice deformation. Ice-sheet
thermal modeling is difficult due to the uncertainties in basal
heat flux (Greve, 2005). Few deep ice cores are available to
validate the models (Alley and others, 1993; Dansgaard and
others, 1993; Greenland Ice Core Project (GRIP) members,
1993; Meese and others, 1994; Dahl-Jensen and others,
1998; NorthGRIP members, 2004). These ice cores provide
temperature measurements through the ice column, but only
in slow-moving areas of the Greenland ice sheet, near ice
divides. Initializing ice-flow models with an appropriate
thermal regime therefore remains challenging, especially for
HO or FS models that cannot run interglacial spin-ups. ð3Þ where
u, v, w
ð
Þ are the three components of velocity in a
Cartesian coordinate system
x, y, z
ð
Þ, with z the vertical
axis, is the ice density, g is the norm of the acceleration
due to gravity and s is the ice upper surface elevation. Vertical velocity is recovered using the equation of
incompressibility. INTRODUCTION p
y
The basal friction follows a viscous law (Cuffey and
Paterson, 2010); the basal condition between the ice and the
underlying bedrock is Here we assess the effect of the initial thermal regime on
century-scale simulations of the Greenland ice sheet. To
address this question, we (1) compare steady-state modeled
ice temperatures against measurements from three deep ice
cores, (2) analyze the sensitivity of inferred properties at the
ice/bedrock interface to changes in ice temperature, (3) as-
sess the effect of initial temperatures on century-scale
projections and (4) compare the ice mass changes due to
different initial temperatures against those obtained by
changing external forcings, such as basal sliding or atmos-
pheric conditions. We conclude by discussing the influence
of the method employed in calculating the thermal regime
on century-scale projections of ice-sheet evolution. n þ 2Nv
k¼ 0,
ð4Þ ð4Þ where is the stress tensor, n is the outward-pointing
normal vector at the base, 2 is the friction coefficient, N is
the effective pressure, v is the basal velocity and
ð Þk is a
projection operator on the tangent to the bedrock. Effective
pressure is N ¼ gh, with h the height of the ice sheet above
hydrostatic equilibrium as a first-order approximation; a
hydrological model would be necessary to obtain a better
estimate of basal pressure. The effective pressure varies with
time as the ice-sheet geometry evolves. Other boundary
conditions include water pressure at the ice front and air
pressure for land-terminating glaciers, non-penetrability of
ice into the underlying bedrock at the ice/bedrock interface
and a free surface at the ice/air interface. Data ð6Þ We use the datasets provided by the SeaRISE assessment
(Bindschadler and others, 2013). The dataset (http://
websrv.cs.umt.edu/isis/index.php/SeaRISE Assessment) in-
cludes mean annual surface temperature and accumulation
from Ettema and others (2009), basal heat flux from Shapiro
and Ritzwoller (2004) and bedrock topography, ice thickness
and surface elevation from Bamber and others (2001). Observed surface velocities needed for the model initializa-
tion are from Rignot (2012). Ice front position is chosen to
match the velocity datasets and is kept fixed with time. We
treat the entire domain as grounded, to be consistent with
the SeaRISE dataset. Large uncertainties exist in the
geothermal heat flux (Greve, 2005; Rogozhina and others,
2012). We therefore adjust the geothermal flux map from
Shapiro and Ritzwoller (2004) with values similar to Greve
(2005) for Dye3 and GRIP. Geothermal flux values of
20 mW m 2 for the Dye3 site and 60 mW m 2 for the GRIP
site are used (Greve, 2005) and combined with the
underlying geothermal heat flux map in their vicinity. We
choose here to use an exponential decay as we move further
from the drilling site, with an influence area of 250 km. where E is the enthalpy, Es is the enthalpy of pure ice, El is
the enthalpy of pure liquid water at the pressure-melting
point, Tpmp, Ki is the ice diffusivity coefficient, Ci is the ice
heat capacity, k ¼ 1 ! ð
Þki þ !kw is the mixture thermal
conductivity (with ! the water fraction and ki and kw the
thermal conductivity of pure ice and liquid water), k0 is a
small positive constant and is the internal deformation
heat. The case of pure water, where enthalpy is above the
enthalpy of pure liquid water, El, is not considered here. py
p
q
The temperature, T, and water fraction, !, are recovered
as follows: For cold ice, E < Es: For cold ice, E < Es: For cold ice, E < Es:
T ¼ E Es
Ci
þ Tpmp
! ¼ 0:
For temperate ice, Es < E < El:
T ¼ Tpmp
! Model set-up To run a large-scale HO model of Greenland and capture
the narrow and fast-flowing ice streams while maintaining a
reasonable computational cost, we rely on anisotropic mesh
adaptation to limit the number of elements. The element size
is optimized to minimize the interpolation error of surface
velocities (Morlighem and others, 2010). The model hori-
zontal resolution varies between 1 km on the fast ice streams
along the coast and 25 km in the slow-moving regions of the
interior, which results in a two-dimensional triangular mesh
of 64 000 elements. The spatial resolution must be
sufficiently high to allow for a good representation of the
outlet glaciers but coarse enough that the model remains
computationally manageable. A 1 km resolution gives a
reasonable trade-off. The horizontal mesh is then extruded
into 25 non-uniform layers between the bedrock and surface
elevations. Vertical grid spacing is refined towards the
bottom where temperature gradients and vertical shearing
are concentrated (the layer next to the bed is half the height
of the upper layer). The 3-D mesh comprises >1500 000 where s is the domain upper surface, b its lower surface,
ðuobs, vobsÞ the observed horizontal velocity and " is a
minimum velocity used to avoid singularities. The first two
terms in this cost function are the L2 misfit, which measures
the square of the difference between model and obser-
vations, and the square of the logarithmic difference between
model and observations. The combination of these two terms
allows best-fitting of observations on both fast-flowing and
slow-moving areas (Morlighem and others, 2010, 2013). The
last term is a Tikhonov regularization term, which penalizes
uncontrolled oscillations of and prevents overfitting the
data. 1,
2 and
t are non-dimensionalizing constants taken
as
1 ¼ 300,
2 ¼ 1:5 and
t ¼ 10 7 in order to balance
these three terms. The algorithm relies on an exact adjoint
that includes dependency of ice viscosity into the strain rate where s is the domain upper surface, b its lower surface,
ðuobs, vobsÞ the observed horizontal velocity and " is a
minimum velocity used to avoid singularities. The first two
terms in this cost function are the L2 misfit, which measures
the square of the difference between model and obser-
vations, and the square of the logarithmic difference between
model and observations. Data ¼ E Es
L
: For the climate scenario (see below), air temperatures and
surface mass balance for the next 94 years follow the A1B
scenario of the Intergovernmental Panel on Climate Change
(IPCC) Fourth Assessment Report (AR4) (Pachauri and
Reisinger, 2007) and are kept constant for the last 6 years
of the run. Surface mass-balance anomalies compared to
present-day values from Ettema and others (2009) are
applied for the climate run, as described by Bindschadler
and others (2013) and Nowicki and others (2013). Air
temperature and surface mass balance are directly applied at
the upper surface elevation. Contrary to the description of Aschwanden and others
(2012b), our model does not include a layer of the Earth’s
lithosphere, so the geothermal heat flux is directly applied at
the ice base; ice in contact with the bedrock is heated by
geothermal and frictional heat flux (Larour and others,
2012a). Air temperature is imposed on the upper surface. Data assimilation of surface velocities is used to infer the
basal friction coefficient as we try to minimize the misfit
between modeled and measured surface velocities. We
define the following cost function, J : Modeled temperatures are compared with measurements
from three deep ice cores: Dye 3 (Gundestrup and Hansen,
1984; Dahl-Jensen and others, 1998); GRIP (Johnsen and
others, 1995; Dahl-Jensen and others, 1998) and Greenland
Ice Sheet Project 2 (GISP2) (Cuffey and others, 1995; Clow
and others, 1996). J ðu, v, Þ ¼
1
1
2
Z
s
u uobs
ð
Þ2þ v vobs
ð
Þ2d s
þ
2
1
2
Z
s
ln
ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
u2 þ v2
p
þ "
ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
uobs2 þ vobs2
p
þ "
!2
d s
þ
t
1
2
Z
b
r r d b
, ð7Þ MODEL AND METHODS
Model description The enthalpy 1026 1026 Seroussi and others: Greenland thermal regime for future projections equation valid for both cold and temperate ice is (Goldberg and Sergienko, 2011) for the derived gradient and
quasi-Newton (limited-memory BFGS (Broyden–Fletcher–
Goldfarb–Shanno); Nocedal, 1980) for the gradient descent. (Goldberg and Sergienko, 2011) for the derived gradient and
quasi-Newton (limited-memory BFGS (Broyden–Fletcher–
Goldfarb–Shanno); Nocedal, 1980) for the gradient descent.
@E
@t þ v rE
¼
þ
r
Ki
Ci
rE
if E < Es
r
krTpmp þ k0
L rE
if Es < E < El
8
>
>
>
<
>
>
>
:
,
ð6Þ MODEL AND METHODS
Model description The evolution of the surface elevation is dictated by mass
conservation. The mass transport equation reads To model the Greenland ice sheet, we use the ice-sheet
system model (ISSM; Morlighem and others, 2010; Seroussi
and others, 2011; Larour and others, 2012a), an open-source
finite-element software that simulates the thermodynamics
of ice sheets at continental scale. Here we summarize the
main characteristics and equations of the models (a
complete description is given by Larour and others, 2012a). Ice is modeled as a viscous incompressible material. We
use the 3-D HO model (Blatter, 1995; Pattyn, 2003) for the
momentum balance equations. This model is derived from
the FS model by making two assumptions: (1) the horizontal
gradients of vertical velocities are negligible compared with
vertical gradients of horizontal velocity and (2) bridging
effects are negligible. This leads to a 3-D model where To model the Greenland ice sheet, we use the ice-sheet
system model (ISSM; Morlighem and others, 2010; Seroussi
and others, 2011; Larour and others, 2012a), an open-source
finite-element software that simulates the thermodynamics
of ice sheets at continental scale. Here we summarize the
main characteristics and equations of the models (a
complete description is given by Larour and others, 2012a). @H
@t ¼ r Hv
ð
Þ þ _Ms _Mb,
ð5Þ ð5Þ where H is the ice thickness, v is the depth-averaged
horizontal velocity, _Ms is the surface mass balance and _Mb is
the basal melting. This hyperbolic equation is stabilized
using a streamline upwinding finite-element method (Brooks
and Hughes, 1982). Ice is modeled as a viscous incompressible material. We
use the 3-D HO model (Blatter, 1995; Pattyn, 2003) for the
momentum balance equations. This model is derived from
the FS model by making two assumptions: (1) the horizontal
gradients of vertical velocities are negligible compared with
vertical gradients of horizontal velocity and (2) bridging
effects are negligible. This leads to a 3-D model where For the thermal model, we use an enthalpy formulation
(as described in Aschwanden and others, 2012b). This
method allows both temperate and cold ice to be
included in an energy-conserving framework easily, as there
is no need to track the cold/temperate transition surface used
in polythermal models (Greve, 1997a,b). Experiments The three different initial thermal regimes (EXP1, EXP2
and EXP3) are then used to assess the sensitivity of inferred
basal friction to initial thermal regime (Fig. 3). All models are
able to accurately reproduce the surface velocities, with an
average misfit of 10.7, 13.5 and 11.0 m a 1 for EXP1, EXP2
and EXP3, respectively. Differences between steady state and
EXP1 are limited, with slightly slower basal velocities (Fig. 3e
and f) and higher friction coefficients in the interior for EXP1
(Fig. 3i and j). This higher friction compensates for vertical
shearing due to higher temperatures next to the base of the
ice sheet. EXP2 and EXP3 initial temperatures are quite
different from the steady-state temperature field so it is not
surprising to see that results of the inversion are also different. EXP2 leads to significantly higher friction coefficients and
lower basal velocities (Fig. 3g and k), especially in the
interior of the ice sheet. Results for EXP3 show the opposite,
i.e. lower friction coefficient and higher basal velocities
(Fig. 3h and l), but the effect is more limited in this case than
for EXP2. Overall, we notice that basal friction and sliding
seem to be more sensitive to initial temperature conditions in
the interior of the ice sheet than in the periphery. We first analyze the effect of initial temperatures on the
inferred basal friction for a static model. We then assess the
sensitivity of the total ice volume to the initial temperature
regimes for projections of 100 years, using constant present-
day conditions of the atmospheric forcing. We rely on four
different initial ice thermal regimes: (1) steady-state tempera-
tures calculated from the initial velocities; (2) a linear
variation from 38C at the bed to 08C at the surface added to
the steady-state temperatures (EXP1); (3) a warm model,
where the temperature varies linearly between the surface
temperature and the pressure-melting point at the base (ice
is therefore at the pressure-melting point everywhere at the
base) (EXP2); and (4) a cold-ice model, where the tempera-
ture is equal to the surface temperature for the entire ice
column (EXP3). Model set-up The combination of these two terms
allows best-fitting of observations on both fast-flowing and
slow-moving areas (Morlighem and others, 2010, 2013). The
last term is a Tikhonov regularization term, which penalizes
uncontrolled oscillations of and prevents overfitting the
data. 1,
2 and
t are non-dimensionalizing constants taken
as
1 ¼ 300,
2 ¼ 1:5 and
t ¼ 10 7 in order to balance
these three terms. The algorithm relies on an exact adjoint
that includes dependency of ice viscosity into the strain rate 1027 Seroussi and others: Greenland thermal regime for future projections We first compare the modeled temperatures (that were
computed assuming thermal steady state) with temperature
measurements along three deep ice cores (Fig. 2). The
modeled steady-state profiles compare well with the meas-
urements. GRIP and GISP2 profiles are similar, as expected,
as these two ice cores are located only 28 km apart. For these
two profiles, modeled ice temperature in the upper 2000 m is
in good agreement with the measurements. Differences up to
58C are concentrated in the lower part of the ice sheet. Differences for Dye3 are smaller, with a maximum difference
of 48C at the base. Standard errors between observed and
modeled temperatures are 1.38C, 1.88C and 1.98C for Dye3,
GISP2 and GRIP ice cores, respectively. prismatic elements. We use the arbitrary Eulerian–
Lagrangian method (Donea and Belytschko, 1992) to update
the mesh at each time-step of the 100 year simulations: the
horizontal mesh is fixed in time while the vertical repartition
of elements varies according to the ice upper surface. g
pp
We initialize the model using the present-day geometry
(ice thickness, surface elevation, ice front position). We use
an inverse method to estimate the basal friction coefficient,
in order to fit the present-day observed surface velocities
(MacAyeal, 1993b; Morlighem and others, 2010). The
thermal regime is assumed to be in steady state, and we
ensure that velocities and temperatures are consistent by
running a thermomechanical steady state until both fields
converge: at each iteration of the inversion, we compute the
ice temperature and update ice hardness accordingly, to
ensure consistent ice flow and thermal regimes (Morlighem
and others, 2010). The initial temperature computed is
therefore based only on present-day conditions and does not
include past climate history. The steady-state temperatures can be compared with
modeled temperatures from paleoclimate reconstructions of
Aschwanden and others (2012) and Rogozhina and others
(2012). Model set-up The pattern of basal temperatures (Fig. 1c) is similar,
with ice at the pressure-melting point over a large portion of
the Greenland ice sheet and colder ice mainly along the
divides and in the north. Experiments In order to compare the effect of the thermal regime with
that of other forcings, we run two additional 100 year
simulations, using steady-state temperatures: (1) a climate
run following the IPCC-AR4 A1B scenario and (2) a sliding
run, in which basal friction is reduced by one-third over the
entire ice sheet, to simulate enhanced basal melting. The
climate experiment simulates realistic changes, and the
sliding experiment a commonly used scenario (e.g. Gillet-
Chaulet and others, 2012; Bindschadler and others, 2013). We then perform four runs of 100 years using these
different initial thermal regimes. Under constant present-day
conditions and with the initial steady-state temperature, the
model yields an increase in ice mass in the next 100 years of
180 Gt a–1, equivalent to an increase of 0.73% of the initial
ice mass of the ice sheet (Fig. 4). Results of Bindschadler
and others (2013) show that mass increase is a limitation
common to about half of the ice-sheet models that
participated in the SeaRISE experiments, the other half
starting from an initial volume that differs from the present-
day volume. Differences between the steady state and EXP1
are limited, with a similar mass gain (0.73% and 0.72%,
respectively). EXP3 mass change is also close to the steady-
state evolution, with a mass gain of 0.75%. Results from
EXP2 are different from the other three, with a lower mass
increase of only 0.62% in 100 years. These results are further
analyzed below. RESULTS The model initial configuration matches the ice geometry
and velocity well, as expected, since it uses present-day
topography and is initialized with data assimilation of
observed velocities. The difference between observed and
modeled velocities is 11.0 m a 1 on average. Figure 1 shows
the initial ice thickness (Fig. 1a) and modeled velocity
(Fig. 1b), as well as the absolute difference between modeled
and observed velocities (Fig. 1d). The largest differences are
found along the periphery of the ice sheet, where velocities
are highest (almost 1000 m a 1 difference on Peterman
Gletscher). Areas with high friction (Fig. 3i) are concentrated
on mountainous regions, while very low friction is found
under fast ice streams, as expected. Additional inversions
were performed with different initial guesses of basal friction
and led to similar results (not shown here), demonstrating the
limited influence of the initial value. Finally, we run scenarios similar to the SeaRISE initiative
(Bindschadler and others, 2013; Nowicki and others, 2013)
with the steady-state temperature field as initial state. Ice
mass change for the climate and sliding scenarios is also
presented in Figure 4. These two scenarios respectively lead
to a mass gain of 0.50% and 0.36%, which is significantly 1028 Seroussi and others: Greenland thermal regime for future projections ial Greenland configuration: (a) ice thickness, H (Bamber and others, 2001), (b) modeled horizontal surface velocity, V
djusted temperature, T, and (d) absolute misfit between observed and modeled horizontal surface velocity overlaid on
Resolution Imaging Spectroradiometer) Mosaic of Greenland. Initial Greenland configuration: (a) ice thickness, H (Bamber and others, 2001), (b) modeled horizontal surface velocity, V, (
d
d
d d
b
l
f
b
b
d
d
d l d h
l
f
l
l d Fig. 1. Initial Greenland configuration: (a) ice thickness, H (Bamber and others, 2001), (b) modeled horizontal surface velocity, V, (c) basal
pressure-adjusted temperature, T, and (d) absolute misfit between observed and modeled horizontal surface velocity overlaid on a MODIS
(Moderate Resolution Imaging Spectroradiometer) Mosaic of Greenland. lower than in all the previous runs. Our 100 year simulations
are therefore significantly more sensitive to the applied
external forcings than the initial temperature. reasonable amount of time (i.e. 4 days for each run of
100 years). RESULTS Previous studies (Hindmarsh, 2004; Gudmunds-
son, 2008; Morlighem and others, 2010) on both idealized
geometries and real glaciers showed that differences
between HO and FS exist, especially in the grounding line
area (Durand and others, 2009; Morlighem and others,
2010) but remain spatially limited, so using HO for our
simulations should not affect our conclusions. Seroussi and others: Greenland thermal regime for future projections Fig. 2. Observed (black curves) and modeled steady-state (green curves) temperature profiles at Dye3, GISP2 and GRIP drilling sites. Initial
temperatures for EXP1 (pink dot-dashed curves), EXP2 (red dashed curves) and EXP3 (blue dotted curves). Fig. 2. Observed (black curves) and modeled steady-state (green curves) temperature profiles at Dye3, GISP2 and GRIP drilling sites. Initial
temperatures for EXP1 (pink dot-dashed curves), EXP2 (red dashed curves) and EXP3 (blue dotted curves). shear; this larger vertical shear is compensated for by a
lower basal velocity in order to obtain the same surface
velocity. This effect is the foundation of the SIA, that is valid
for most ice-sheet interiors, but does not hold for fast-
flowing outlet glaciers, where sliding dominates ice motion. modeled steady-state temperatures match the observations
along the three deep ice cores reasonably well. The main
features of the temperature profiles (e.g. surface tempera-
ture, basal temperature, basal temperature gradient) are
captured, with a standard error varying between 1.3 and
1.98C, even though differences with deep ice cores reach
58C near the glacier base. These errors correspond to an
average ice rigidity difference of 3.5% for Dye3 and 5% for
GRIP and GISP. In the upper part of the ice, the modeled
temperatures reach a constant value, which is not consistent
with the observations. This is due to the air temperature
history that is not captured here, as opposed to Greve (2005)
and Rogozhina and others (2012): the temperature decrease
caused by the Last Glacial Maximum is not captured by our
steady-state method. Paleoclimate reconstructions, which
account for temporal changes in surface conditions, yield a
better agreement between modeled temperature and meas-
urements along deep ice core (e.g. <38C in Rogozhina and
others, 2012). It is important to note that these deep ice-core
drillings were collected near ice divides, where conduction
is dominant, and might not be representative of areas where
advection is dominant. Comparisons in more dynamic areas
would better inform the steady-state model’s ability to
capture variations, but measurements in these regions are
almost non-existent. The pattern of inferred basal friction based on steady-state
temperatures is consistent with the results of Larour and
others (2012a) and Gillet-Chaulet and others (2012) for
Greenland, and Joughin and others (2001) and Seroussi and
others (2011) for the northeast Greenland ice stream, with
motion mainly due to sliding on fast-flowing areas and
differences from observations concentrated on the fast-
flowing outlet glaciers. DISCUSSION Large-scale high-resolution simulations require intensive
computational resources. In this study, we use a 3-D HO
model, as a trade-off between first-order SSA/SIA and FS. Our model comprises 1500 000 elements to capture outlet
glaciers while running century-scale simulations in a The numerical experiments performed here show that the
assumption of thermal steady state is a viable approximation
for short-term simulations of the Greenland ice sheet. First, Seroussi and others: Greenland thermal regime for future projections 1029 Seroussi and others: Greenland thermal regime for future projections These models use different approx-
imations of FS and different temperature fields (thermal
steady state for Larour and others, 2012a; an interpolation
from a paleoclimate reconstruction for Gillet-Chaulet and
others, 2012) and confirm that ice temperatures have little
impact on the basal friction inferred from surface velocities. Finally, the projection of the ice-sheet mass balance over
the next 100 years shows an increase in mass of 0.73%. This result is not consistent with the current trends that show
a rapid and accelerated decrease of the Greenland ice sheet
mass (e.g. Rignot and others, 2011). However, a mass
increase is not unusual for ice-sheet models of the Green-
land ice sheet. About half the models that participated in the
Greenland SeaRISE experiments show a mass increase for
the control run that keeps present-day conditions constant
(Bindschadler and others, 2013), although they all use
different ice-flow approximations, spatial resolutions and
initialization procedures. The other models show a mass
decrease but start from an initial volume that differs from the
actual volume. Mass increase often happens for models
initialized with inverse methods and is caused by data that
have inconsistencies (Seroussi and others, 2011), incorrect
position of the ice-sheet margin, mesh resolutions too
coarse (Gillet-Chaulet and others, 2012) and/or missing
physical processes, such as ice/ocean interaction and basal Second, for all four temperature fields the basal drag
inversions yield a good agreement between modeled and
observed velocities. The temperature field affects the friction
coefficient pattern, especially with the extreme cold and
warm cases (EXP2 and EXP3), but the basal stress remains
similar in all four cases. This result corroborates the
conclusions of Scha¨fer and others (2012), who showed that
the friction distribution of Vestfonna ice cap, Nord-
austlandet/Svalbard, does not depend significantly on ice
temperature. The surface velocity is optimized to fit obser-
vations and should not vary from one simulation to the next. Warmer ice deforms more easily and leads to more vertical 1030 Seroussi and others: Greenland thermal regime for future projections i
(
d) S
f
l
i i
(
h) b
l
l
i i
d (i l) f i i
ffi i
2
f
h
f
fil
i
l Fig. 3. (a–d) Surface velocities, (e–h) basal velocities and (i–l) friction coefficient, 2, for the four temperature profiles. Seroussi and others: Greenland thermal regime for future projections This is not the case for long interglacial
runs, where temperature plays a major role in ice-sheet
growth; Greve (2005) and Rogozhina and others (2012)
noticed that small variations in geothermal flux led to
differences in ice volume of up to 10%. By contrast, changes
in external forcings, such as atmospheric conditions or basal
friction, lead to significant variations in ice-sheet mass over
100 years, despite the moderate forcings applied in our
forcing experiments. Results from the SeaRISE experiments
(Bindschadler and others, 2013) and Gillet-Chaulet and
others (2012) also show the strong influence of these
forcings over short timescales. Fig. 4. Simulated ice mass change of the Greenland ice sheet (in Gt
and %) for 100 year simulations with different scenarios: constant
climate with steady-state temperature (green), EXP1 temperature
(violet), EXP2 temperature (red), EXP3 temperature (dark blue),
AR4 atmospheric conditions (yellow) and increased basal lubrica-
tion (gray). higher for EXP2 than for the steady state and EXP1 and EXP3
cases. The inversion for EXP2 did not converge as well as the
others, even though the absolute misfit is only slightly higher
for this experiment than for the others (13.5 m a 1 for EXP2
vs 10.7 and 11.0 m a 1 for EXP1 and EXP3). This suggests
that the model is highly sensitive to the initial modeled
velocity and that this parameter has a significantly larger
impact on simulations than changes in initial temperatures. The basal friction law employed here is not temperature-
dependent, and only the effective pressure changes as the
ice-sheet geometry evolves. Projections of the Greenland ice
sheet evolution could be more sensitive to the initial
temperature field if a temperature-dependent friction law
were used, as nonlinear effects could develop. However,
most of the ice sheet is already at the pressure-melting point
(Fig. 1c), which limits the impact of such a law, and our
conclusions should not be significantly affected. A basal
hydrological model that models effective pressure and water
content (de Fleurian, 2013) is under development and will
improve this aspect in the future. g
As mentioned in the previous section, mass increase is
higher for EXP3 than for the steady-state experiment and
lower for EXP1 and EXP2. Seroussi and others: Greenland thermal regime for future projections First column
corresponds to steady-state temperature (a, e, i), second column to EXP1 (b, f, j), third column to EXP2 (c, g, k) and fourth column to EXP3
(d, h, l). Seroussi and others: Greenland thermal regime for future projections 1031 Fig. 4. Simulated ice mass change of the Greenland ice sheet (in Gt
and %) for 100 year simulations with different scenarios: constant
climate with steady-state temperature (green), EXP1 temperature
(violet), EXP2 temperature (red), EXP3 temperature (dark blue),
AR4 atmospheric conditions (yellow) and increased basal lubrica-
tion (gray). hydrology. Models by Seddik and others (2012) and Gillet-
Chaulet and others (2012) show a volume increase of 0.81%
and 0.28% in 100 years, respectively, after an initial
relaxation of 100 and 50 years, respectively. Such a
relaxation allows the reconciliation of inconsistent model
inputs that lead to unphysical flux divergences (Seroussi and
others, 2011) and limit the high rates of thickness change. However, no ice-sheet model is able to replicate the present-
day configuration and evolution today. We did not use such a
relaxation technique here, which could lead to slightly
different initial conditions (ice thickness, initial volume,
driving stress, etc.), in order to ensure that the ice-sheet
response is due only to changes in the ice thermal regime. For century-scale projections, our experiments show that
the initial thermal regime has only a limited effect on the
evolution of the Greenland ice sheet. Ice temperature of
models that do not start with a thermal steady state see their
temperatures gradually adjusted, but this is a slow process
that has a limited effect for short-term simulations. Influence
of geothermal heat flux will have a similar impact. Larour
and others (2012b) showed that large variations of geother-
mal flux affected the ice rigidity by up to 5% in slow-moving
areas, where basal friction and geothermal energy are the
same order of magnitude. This is within the range of the
initial temperature variations that we tested here. We
therefore expect that uncertainties in geothermal heat flux
do not significantly affect the behavior of the ice sheet for
century-scale simulations. This is consistent with the results
of Larour and others (2012b), who concluded that un-
certainties in geothermal heat flux affect glacier mass
balance by <1%. Seroussi and others: Greenland thermal regime for future projections We would expect the opposite: for
identical surface velocities, depth-averaged velocities
should be smaller for warmer ice, as there is more vertical
shear and basal velocity from the inversion shows a lower
basal velocity for EXP2 than for the steady-state case. Ice
discharge is mainly controlled by fast-flowing outlet gla-
ciers, and as motion due to vertical shear only represents a
fraction of the ice velocity in these areas, ice flux is not
affected. Conversely, differences between observed and
modeled velocities have a stronger effect. Contrary to
EXP1 and EXP3, Greenland mass evolution with EXP2 is
quite different from the steady-state case. This is due to the
initial velocity after inversion, which is higher than
observed, especially in the eastern and southeastern part
of Greenland (Fig. 5a), where the difference reaches
50 m a 1 over large areas. In all other cases, the modeled
velocity is usually smaller than the measurements (e.g. EXP3
in Fig. 5b: the difference between observed and modeled
velocities is 0 m a 1 in those same areas). This higher
velocity leads to a discharge at the ice front that is 10% Realistic modeling of the ice thermal regime remains a
major objective and more in situ measurements need to be
acquired to calibrate and validate models. There are few
temperature measurements available in fast-flowing areas,
and uncertainties associated with the geothermal heat flux,
which is a key control on the thermal regime, remain large. Our results indicate that variations of <58C in parts of the ice
sheet in the initial temperature field have a limited effect on
both initializations with data assimilation and ice-sheet
evolution for short-term simulations of large-scale models. Other factors and parameters are of greater importance: an
accurate representation of small outlet glaciers is necessary
to capture the ice discharge; a realistic description of the
ice-sheet present-day configuration with improved bedrock
topography data is needed to reduce uncertainties, as well as
better constrain mass flux and unknown parameters (Ser-
oussi and others, 2011; Gillet-Chaulet and others, 2012). These aspects are more limiting for ice-sheet numerical
modeling and affect sea-level rise projections at the contin-
ental scale more significantly than small variations in the ice
thermal regime. 1032 Seroussi and others: Greenland thermal regime for future projections Fig. 5. Modeled minus observed velocities for (a) EXP2 and (b) EXP3. Modeled velocities are higher than observations in red areas and lower
in blue areas. Fig. 5. Seroussi and others: Greenland thermal regime for future projections Modeled minus observed velocities for (a) EXP2 and (b) EXP3. Modeled velocities are higher than observations in red areas and lower
in blue areas. REFERENCES Alley RB and 10 others (1993) Abrupt increase in Greenland snow
accumulation at the end of the Younger Dryas event. Nature,
362(6420), 527–529 In this study, we have presented simulations performed with
a 3-D thermomechanical HO model of the Greenland ice
sheet. Anisotropic mesh adaptation and data assimilation
allow the numerical model to capture outlet glaciers with
better accuracy and to match the present-day conditions in a
realistic manner. We show that modeled temperatures,
assuming an ice sheet in thermal steady state, are in
agreement with in situ measurements along three deep
ice-core profiles. The impact on ice rigidity is 5% or less,
and such changes do not have a significant influence on
basal friction inversion and ice-sheet evolution for century-
scale projections. The ice-sheet model is far more sensitive
to changes in external forcings, namely basal sliding and
atmospheric conditions, than its initial temperature. Using a
steady-state approach combined with present-day condi-
tions to compute the ice thermal regime is a viable
alternative to spin-up when looking at short-term simula-
tions of the Greenland ice sheet at the continental scale. Applegate PJ, Kirchner N, Stone EJ, Keller K and Greve R (2012) An
assessment of key model parametric uncertainties in projections Applegate PJ, Kirchner N, Stone EJ, Keller K and Greve R (2012) An
assessment of key model parametric uncertainties in projections
of Greenland Ice Sheet behavior. Cryosphere, 6(3), 589–606
(doi: 10.5194/tc-6-589-2012) of Greenland Ice Sheet behavior. Cryosphere, 6(3), 589–606
(doi: 10.5194/tc-6-589-2012) Aschwanden A, Aðalsgeirsdo´ttir G and Khroulev C (2012a)
Hindcasting to measure ice sheet model sensitivity to initial
states. Cryos. Discuss., 6(6), 5069–5094 (doi: 10.5194/tcd-6-
5069-2012) Aschwanden A, Bueler E, Khroulev C and Blatter H (2012b) An
enthalpy formulation for glaciers and ice sheets. J. Glaciol., 58(209), 441–457 (doi: 10.3189/2012JoG11J088) Bamber JL, Layberry RL and Gogineni SP (2001) A new ice
thickness and bed data set for the Greenland ice sheet. 1. Measurement, data reduction, and errors. J. Geophys. Res.,
106(D24), 33 773–33 780 (doi: 10.1029/2001JD900054) Benn DI, Warren CW and Mottram RH (2007) Calving processes
and the dynamics of calving glaciers. Earth-Sci. Rev., 82(3–4),
143–179 (doi: 10.1016/j.earscirev.2007.02.002) Bindschadler RA and 27 others (2013) Ice-sheet model sensitivities
to environmental forcing and their use in projecting future sea
level (the SeaRISE project). J. Glaciol., 59(214), 195–224 (doi:
10.3189/2013JoG12J125) ACKNOWLEDGEMENTS Tellus, 47B(5),
624–629 Dansgaard W and 10 others (1993) Evidence for general instability
of past climate from a 250 kyr ice-core record. Nature,
364(6434), 218–220 (doi: 10.1038/364218a0) Joughin I, Fahnestock M, MacAyeal D, Bamber JL and Gogineni P
(2001) Observation and analysis of ice flow in the largest Green-
land ice stream. J. Geophys. Res., 106(D24), 34 021–34 034
(doi: 10.1029/2001JD900087) De Fleurian B and 6 others (2013) A subglacial hydrological model
dedicated to glacier sliding. Cryos. Discuss., 7(4), 3449–3496
(doi: 10.5194/tcd-7-3449-2013) Larour E, Seroussi H, Morlighem M and Rignot E (2012a)
Continental scale, high order, high spatial resolution, ice sheet
modeling using the Ice Sheet System Model (ISSM). J. Geophys. Res., 117(F1), F01022 (doi: 10.1029/2011JF002140) DeConto RM and Pollard D (2003) Rapid Cenozoic glaciation of
Antarctica induced by declining atmospheric CO2. Nature,
421(6920), 245–248 (doi: 10.1038/nature01290) Donea J and Belytschko T (1992) Advances in computational
mechanics. Nucl. Eng. Des., 134(1), 1–22 (doi: 10.1016/0029-
5493(92)90004-F) Larour E, Morlighem M, Seroussi H, Schiermeier J and Rignot E
(2012b) Ice flow sensitivity to geothermal heat flux of Pine Island
Glacier, Antarctica. J. Geophys. Res., 117(F4), F04023 (doi:
10.1029/2012JF002371) Durand G, Gagliardini O, Zwinger T, Le Meur E and Hindmarsh RCA
(2009) Full Stokes modeling of marine ice sheets: influence of the
grid size. Ann. Glaciol., 50(52), 109–114 (doi: 10.3189/
172756409789624283) MacAyeal DR (1989) Large-scale ice flow over a viscous basal
sediment: theory and application to Ice Stream B, Antarctica. J. Geophys. Res., 94(B4), 4071–4087 (doi: 10.1029/
JB094iB04p04071) Ettema J and 6 others (2009) Higher surface mass balance of the
Greenland ice sheet revealed by high-resolution climate model-
ling. Geophys. Res. Lett., 36(12), L12501 (doi: 10.1029/
2009GL038110) MacAyeal DR (1993a) A tutorial on the use of control methods in
ice-sheet modeling. J. Glaciol., 39(131), 91–98 MacAyeal DR (1993b) Binge/purge oscillations of the Laurentide
ice sheet as a cause of the North Atlantic’s Heinrich events. Paleoceanography, 8(6), 775–784 (doi: 10.1029/93PA02200) Gillet-Chaulet F and 8 others (2012) Greenland Ice Sheet contri-
bution to sea-level rise from a new-generation ice-sheet model. Cryosphere, 6(4), 1561–1576 (doi: 10.5194/tc-6-1561-2012) McFadden EM, Howat IM, Joughin I, Smith BE and Ahn Y (2011)
Changes in the dynamics of marine terminating outlet glaciers in
west Greenland (2000–2009). J. Geophys. Res., 116(F2), F02022
(doi: 10.1029/2010JF001757) Glen JW (1955) The creep of polycrystalline ice. Proc. R. Soc. London, Ser. A, 228(1175), 519–538 (doi: 10.1098/rspa. ACKNOWLEDGEMENTS This work was performed at the Jet Propulsion Laboratory
(JPL), California Institute of Technology, and University of
California, Irvine, under a contract with the NASA Cryo-
spheric Sciences Program. H.S. was supported by an
appointment to the NASA postdoctoral program at the JPL,
administered by Oak Ridge Associated Universities through
a contract with NASA. A.K. was supported by a grant from
NASA’s Cryospheric Sciences Program. We thank the scien-
tific editor, R. Greve, as well as A. Aschwanden and an
anonymous reviewer for comments which helped improve
the clarity of the manuscript. Blatter H (1995) Velocity and stress fields in grounded glaciers: a
simple algorithm for including deviatoric stress gradients. J. Glaciol., 41(138), 333–344 Brooks AN and Hughes TJR (1982) Streamline upwind/Petrov–
Galerkin formulations for convection dominated flows with
particular emphasis on the incompressible Navier–Stokes equa-
tions. Comput. Meth. Appl. Mech. Eng., 32(1–3), 199–259 (doi:
10.1016/0045-7825(82)90071-8) Bueler E and Brown J (2009) Shallow shelf approximation
as a ‘sliding law’ in a thermomechanically coupled ice sheet 1033 Seroussi and others: Greenland thermal regime for future projections modeling. J. Geophys. Res., 109(F1), F01012 (doi: 10.1029/
2003JF000065) modeling. J. Geophys. Res., 109(F1), F01012 (doi: 10.1029/
2003JF000065) model. J. Geophys. Res., 114(F3), F03008 (doi: 10.1029/
2008JF001179) model. J. Geophys. Res., 114(F3), F03008 (doi: 10.1029/
2008JF001179) Clow GD, Saltus RW and Waddington ED (1996) A new high-
precision borehole-temperature logging system used at GISP2,
Greenland, and Taylor Dome, Antarctica. J. Glaciol., 42(142),
576–584 Holland DM, Thomas RH, de Young B, Ribergaard MH and
Lyberth B (2008) Acceleration of Jakobshavn Isbr triggered by
warm subsurface ocean waters. Nature Geosci., 1(10), 659–664
(doi: 10.1038/ngeo316) Cuffey KM and Paterson WSB (2010) The physics of glaciers, 4th
edn. Butterworth-Heinemann, Oxford Howat IM, Joughin IR and Scambos TA (2007) Rapid changes in ice
discharge from Greenland outlet glaciers. Science, 315(5818),
1559–1561 (doi: 10.1126/science.1138478) Cuffey KM, Clow GD, Alley RB, Stuiver M, Waddington ED and
Saltus RW (1995) Large Arctic temperature change at the
Wisconsin–Holocene glacial transition. Science, 270(5235),
455–458 (doi: 10.1126/science.270.5235.455) Hutter K (1982) Dynamics of glaciers and large ice masses. Annu. Rev. Fluid Mech., 14, 87–130 (doi: 10.1146/annurev.fl.14. 010182.000511) Dahl-Jensen D and 6 others (1998) Past temperatures directly from
the Greenland ice sheet. Science, 282(5387), 268–271 (doi:
10.1126/science.282.5387.268) Johnsen SJ, Dahl-Jensen D, Dansgaard W and Gundestrup NS
(1995) Greenland paleotemperatures derived from GRIP bore-
hole temperature and ice core isotope profiles. ACKNOWLEDGEMENTS 1955.0066) Goldberg DN and Sergienko OV (2011) Data assimilation using a
hybrid ice flow model. Cryosphere, 5(2), 315–327 (doi:
10.5194/tc-5-315-2011) Meese DA and 8 others (1994) The accumulation record from the
GISP2 core as an indicator of climate change throughout the
Holocene. Science, 266(5191), 1680–1682 (doi: 10.1126/
science.266.5191.1680) Greenland Ice-Core Project (GRIP) members (1993) Climate instabil-
ity during the last interglacial period recorded in the GRIP ice
core. Nature, 364(6434), 203–207 (doi: 10.1038/364203a0) Moon T, Joughin I, Smith B and Howat I (2012) 21st-century
evolution of Greenland outlet glacier velocities. Science,
336(6081), 576–578 (doi: 10.1126/science.1219985) Greve R (1997a) A continuum-mechanical formulation for shallow
polythermal ice sheets. Philos. Trans. R. Soc. London, Ser. A,
355(1726), 921–974 (doi: 10.1098/rsta.1997.0050 ) Morlighem M, Rignot E, Seroussi H, Larour E, Ben Dhia H and
Aubry D (2010) Spatial patterns of basal drag inferred using
control methods from a full-Stokes and simpler models for Pine
Island Glacier, West Antarctica. Geophys. Res. Lett., 37(14),
L14502 (doi: 10.1029/2010GL043853) Greve R (1997b) Application of a polythermal three-dimensional
ice sheet model to the Greenland ice sheet: response to steady-
state and transient climate scenarios. J. Climate, 10(5), 901–918
(doi: 10.1175/1520-0442(1997)010<0901:AOAPTD>2.0.CO;2) Morlighem M, Seroussi H, Larour E and Rignot E (2013) Inversion of
basal friction in Antarctica using exact and incomplete adjoints
of a higher-order model. J. Geophys. Res., 118(2) (doi: 10.1002/
jgrf.20125) Greve R (2005) Relation of measured basal temperatures and the
spatial distribution of the geothermal heat flux for the Greenland
ice sheet. Ann. Glaciol., 42(1), 424–432 (doi: 10.3189/
172756405781812510) Nocedal J (1980) Updating quasi-Newton matrices with limited
storage. Math. Comput., 35(151), 773–782 (doi: 10.1090/
S0025-5718-1980-0572855-7) Greve R, Saito F and Abe-Ouchi A (2011) Initial results of the
SeaRISE numerical experiments with the models SICOPOLIS
and IcIES for the Greenland ice sheet. Ann. Glaciol., 52(58),
23–30 (doi: 10.3189/172756411797252068) North Greenland Ice Core Project (NorthGRIP) members (2004)
High-resolution record of Northern Hemisphere climate
extending into the last interglacial period. Nature, 431 (7005),
147–151 (doi: 10.1038/nature02805) 23 30 (doi: 10.3189/172756411797252068)
Gudmundsson GH (2008) Analytical solutions for the surface
response to small amplitude perturbations in boundary data in
the shallow-ice-stream approximation. Cryosphere, 2(2), 77–93
(doi: 10.5194/tc-2-77-2008) 147–151 (doi: 10.1038/nature02805) Nowicki S and 30 others (2013) Insights into spatial sensitivities of
ice mass response to environmental change from the SeaRISE ice
sheet modeling project I: Antarctica. J. Geophys. ACKNOWLEDGEMENTS Res., 118(F2),
1002–1024 (doi: 10.1002/jgrf.20081) Gundestrup NS and Hansen BL (1984) Bore-hole survey at Dye 3,
south Greenland. J. Glaciol., 30(106), 282–288 jg
Pachauri RK and Reisinger A eds. (2007) Climate change 2007:
Synthesis Report. Contribution of Working Groups I, II and III to Hindmarsh RCA (2004) A numerical comparison of approx-
imations to the Stokes equations used in ice sheet and glacier 1034 Seroussi and others: Greenland thermal regime for future projections for altitude changes in the Vostok region. J. Geophys. Res.,
106(D23), 31 943–31 964 (doi: 10.1029/2001JD900232) for altitude changes in the Vostok region. J. Geophys. Res.,
106(D23), 31 943–31 964 (doi: 10.1029/2001JD900232) the Fourth Assessment Report of the Intergovernmental Panel
on Climate Change. Intergovernmental Panel on Climate
Change, Geneva Rogozhina I and 6 others (2012) Effects of uncertainties in the
geothermal heat flux distribution on the Greenland Ice Sheet: an
assessment of existing heat flow models. J. Geophys. Res.,
117(F2), F02025 (doi: 10.1029/2011JF002098) Pattyn F (2003) A new three-dimensional higher-order thermo-
mechanical ice-sheet model: basic sensitivity, ice stream
development, and ice flow across subglacial lakes. J. Geophys. development, and ice flow across subglacial lakes. J. Geophys. Res., 108(B8), 2382 (doi: 10.1029/2002JB002329) Scha¨fer M and 8 others (2012) Sensitivity of basal conditions in an
inverse model: Vestfonna ice cap, Nordaustlandet/Svalbard. Cryosphere, 6(4), 771–783 (doi: 10.5194/tc-6-771-2012) Phillips T, Rajaram H and Steffen K (2010) Cryo-hydrologic
warming: a potential mechanism for rapid thermal response of
ice sheets. Geophys. Res. Lett., 37(20), L20503 (doi: 10.1029/
2010GL044397) Schoof C (2010) Ice-sheet acceleration driven by melt supply
variability. Nature, 468(7325), 803–806 (doi: 10.1038/
nature09618) Pollard D and DeConto RM (2009) Modelling West Antarctic ice
sheet growth and collapse through the past five million years. Nature, 458(7236), 329–332 (doi: 10.1038/nature07809) Seddik H, Greve R, Zwinger T, Gillet-Chaulet F and Gagliardini O
(2012) Simulations of the Greenland ice sheet 100 years into the
future with the full Stokes model Elmer/Ice. J. Glaciol., 58(209),
427–440 (doi: 10.3189/2012JoG11J177) Price SF, Payne AJ, Howat IM and Smith BE (2011) Committed sea-
level rise for the next century from Greenland ice sheet
dynamics during the past decade. Proc. Natl Acad. Sci. USA
(PNAS), 108(22), 8978–8983 (doi: 10.1073/pnas.1017313108) Seroussi H and 6 others (2011) Ice flux divergence anomalies on
79north Glacier, Greenland. Geophys. Res. Lett., 38(9), L09501
(doi: 10.1029/2011GL047338) Rignot E (2012) Ice flow in Greenland for the International Polar
Year 2008–2009. Geophys. Res. ACKNOWLEDGEMENTS Lett., 39(11), L11501 (doi:
10.1029/2012GL051634) Shapiro NM and Ritzwoller MH (2004) Inferring surface heat flux
distribution guided by a global seismic model: particular
application to Antarctica. Earth Planet. Sci. Lett., 223(1–2),
213–224 (doi: 10.1016/j.epsl.2004.04.011) Rignot E, Velicogna I, Van den Broeke MR, Monaghan A and
Lenaerts J (2011) Acceleration of the contribution of the
Greenland and Antarctic ice sheets to sea level rise. Geophys. Res. Lett., 38(5), L05503 (doi: 10.1029/2011GL046583) (
j p
)
Velicogna I (2009) Increasing rates of ice mass loss from the
Greenland and Antarctic ice sheets revealed by GRACE. Geophys. Res. Lett.,
36(19),
L19503 (doi:
10.1029/
2009GL040222) Ritz C, Rommelaere V and Dumas C (2001) Modeling the evolution
of Antarctic ice sheet over the last 420 000 years: implications MS received 19 March 2013 and accepted in revised form 23 August 2013
|
https://openalex.org/W2997184163
|
https://europepmc.org/articles/pmc7017149?pdf=render
|
English
| null |
Structure–Function Relationship and Physiological Roles of Transient Receptor Potential Canonical (TRPC) 4 and 5 Channels
|
Cells
| 2,019
|
cc-by
| 14,500
|
Structure–Function Relationship and Physiological
Roles of Transient Receptor Potential Canonical
(TRPC) 4 and 5 Channels Jinsung Kim 1,†
, Juyeon Ko 1,†, Chansik Hong 2,† and Insuk So 1,* Jinsung Kim 1,†
, Juyeon Ko 1,†, Chansik Hong 2,† and Insuk So 1,*
1
Department of Physiology, College of Medicine, Seoul National University, Seoul 03080, Korea;
jinsung_kim@snu.ac.kr (J.K.); juuyeon@snu.ac.kr (J.K.)
2
Department of Physiology, College of Medicine, Chosun University, Gwangju 61452, Korea;
cshong@chosun.ac.kr Jinsung Kim 1,†
, Juyeon Ko 1,†, Chansik Hong 2,† and Insuk So 1,*
1
Department of Physiology, College of Medicine, Seoul National University, Seoul 03080, Korea;
jinsung_kim@snu.ac.kr (J.K.); juuyeon@snu.ac.kr (J.K.)
2
Department of Physiology, College of Medicine, Chosun University, Gwangju 61452, Korea;
cshong@chosun.ac.kr g
*
Correspondence: insuk@snu.ac.kr; Tel.: +82-2-740-8228 †
These authors contributed equally to this work. Received: 30 November 2019; Accepted: 24 December 2019; Published: 27 December 2019 Abstract: The study of the structure–function relationship of ion channels has been one of the
most challenging goals in contemporary physiology. Revelation of the three-dimensional (3D)
structure of ion channels has facilitated our understanding of many of the submolecular mechanisms
inside ion channels, such as selective permeability, voltage dependency, agonist binding, and
inter-subunit multimerization. Identifying the structure–function relationship of the ion channels
is clinically important as well since only such knowledge can imbue potential therapeutics with
practical possibilities. In a sense, recent advances in the understanding of the structure–relationship
of transient receptor potential canonical (TRPC) channels look promising since human TRPC
channels are calcium-permeable, non-selective cation channels expressed in many tissues such as
the gastrointestinal (GI) tract, kidney, heart, vasculature, and brain. TRPC channels are known
to regulate GI contractility and motility, pulmonary hypertension, right ventricular hypertrophy,
podocyte injury, seizure, fear, anxiety-like behavior, and many others. In this article, we tried to
elaborate recent findings of Cryo-EM (cryogenic-electron microscopy) based structural information of
TRPC 4 and 5 channels and domain-specific functions of the channel, such as G-protein mediated
activation mechanism, extracellular modification of the channel, homo/hetero-tetramerization, and
pharmacological gating mechanisms. Keywords: structure–function relationship; transient receptor potential canonical; TRPC cells cells 1. Introduction Transient receptor potential canonical (TRPC) channels constitute non-selective, Ca2+-permeable
channels in mammalian cells [1–3]. There are 7 TRPC subtypes, namely, TRPC1–7, where 6 subtypes
except for TRPC2 can be found in mammalian cells [4]. Based on amino acid sequence similarities,
TRPC1, 4, and 5 channels are grouped as one subfamily and TRPC 3, 6, 7 are grouped as the other [5,6]. The expression pattern of TRPC1 in mammalian tissue is ubiquitous. TRPC4 and 5, however, show
somewhat localized expression profile such as brain [7–18], gastrointestinal tract (GI tract) [2,3,19–22],
ovary [23,24], endometrium [25,26], ventricular myocytes [27–29], and vasculature [30–33]. In such
tissues, TRPC 1, 4, and 5 channels are known to modulate fear or stress response, integrity of
dopaminergic motor neurons such as substantia nigra, hypertrophic cardiomyopathy, endothelial
dysfunction, cholinergic contraction of GI smooth muscle, and many others. Because of the overlapping
expression pattern and capability for heteromerization, channel composition in the plasma membrane Cells 2020, 9, 73; doi:10.3390/cells9010073 www.mdpi.com/journal/cells www.mdpi.com/journal/cells Cells 2020, 9, 73 2 of 24 of native tissues is usually heterogeneous; TRPC4/4, TRPC5/5, TRPC1/4 or TRPC1/5 channels can all
be found. ative tissues is usually heterogeneous; TRPC4/4, TRPC5/5, TRPC1/4 or TRPC1/5 channels can al
ound. of native tissues is usually heterogeneous; TRPC4/4, TRPC5/5, TRPC1/4 or TRPC1/5 channels can all
b f
d Classically, TRPC4 and 5 channels are known to be activated by G-proteins and their downstream
signal transduction pathways, such as Gq-PLC pathway [2,3,21,22,34,35]. Intensive researches have
been done on delineating functional relationship among TRPC4, 5 channels and phosphatidylinositol
4,5-bisphosphate (PIP2) [36–42], intracellular Ca2+ ([Ca2+]i) and protein kinase C (PKC) [43,44]. Recently,
it was suggested that not only downstream signal transduction pathways but active form of alpha
subunits of G-proteins (Gαi or Gαq) per se can activate TRPC4, TRPC5, TRPC1/4, or TRPC1/5 channels
as well [38,39,45,46]. Earlier studies of TRPC channels have gone through some struggles due to
lack of channel-specific agonists or antagonists. In a sense, a recent development of channel-specific
pharmacological tools such as ML-204 [47], (-)-Englerin-A (or simply, Englerin-A) [48] and Pico-145 [49]
has brought great utility to the TRPC community. With such pharmacological treasures, dissecting each
contribution of the TRPC channels among the mixture of channels in plasma membrane is becoming
clearer than ever. In spite of functional utility with which the channel-specific drugs invest, however,
study for mechanism of action of those drugs with structural insight is unfortunately limited. 1. Introduction The great technological advance in the Cryo-EM ‘resolution-revolution’ has helped in revealing
atomic structures [50] of many transmembrane proteins, not to mention TRPC channels [51–53]. Meanwhile, there has been more than 30 years of discussion about functionality of the TRPC channels. Possible molecular candidates that might affect opening or closing of the channel had been numerously
reported, and the domains or amino acids within the channel that might be responsible for the
interaction with such molecular candidates have been rigorously found. In this article, we first tried to reorganize some critical results in functional studies of TRPC1,
4, or 5 channels based on our expertise. As a next step, we elaborated a possible structure–function
relationship in each topic based on the Cryo-EM structure of the channels. Finally, we conclude this
article with challenging questions that still have not been answered even with the Cryo-EM structure. We presume that such questions should be answered in order to fully understand TRPC4, 5, and related
heterotetrameric channels (TRPC1/4 and TRPC1/5). 2. General Architecture of TRPC4 and TRPC5 Channels Like many of voltage-dependent ion channels, TRPC channels share a common architectural
framework of such channels. TRPC channels have six transmembrane segments and two large
cytoplasmic domains, namely, the N-terminal domain and C-terminal domain. At N-terminus, there
are 4 ankyrin-repeat domains (ARD) which have crucial roles in subunit–subunit multimerization,
membrane trafficking, and proper protein folding. Between ARD and the first transmembrane segment
(S1), there exists a helical domain whose axis is somewhat parallel to the inner border of the plasma
membrane: ‘pre-S1 helix’. Between the pre-S1 helix and ARD, 7 α-helices connected with loops can be
found. These are called the ‘helix-loop-helix (HLH)’ domain. The last component in the N-terminal
cytoplasmic domains is the ‘pre-S1 elbow’ which is between the HLH and pre-S1 helix. After the
pre-S1 helix, four transmembrane segments (S1-S4) constitute the voltage-sensor like domain (VSLD)
of the TRPC channels whose functional role and topology resemble the voltage-sensor domain of the
voltage-gated potassium channels [54]. The VSLD of TRPC4 and 5 channels share great similarity with
the VSLD of TRPC3 or TRPC6 channels except S3 [55,56]. S3 of the TRPC3 and TRPC6 channels are
longer than S3 of the TRPC4 and TRPC5 channels, protruding beyond the extracellular borderline of
the plasma membrane. The difference in S3 may reflect some possible discrepancies in extracellular
channel modulation mechanisms between TRPC3, 6, and TRPC4, and 5 channels. The fifth and sixth
transmembrane segment (S5, S6), the extracellular linker sequence between S5 and S6 (S5-S6 linker) and
helical structure within the S5-S6 linker (‘pore helix’), constitute the core framework for the central ion
conduction pathway in a domain-swapped fashion. After S6, a long C-terminal cytoplasmic domain
unfolds where a hydrophobic helical domain comes first. This hydrophobic helical domain has a
TRP signature sequence (EWKFAR), hence called ‘TRP helix’ or ‘TRP box’. The TRP sequence can 3 of 24 Cells 2020, 9, 73 be found not only in TRPC channels but in the TRPM and TRPV superfamily. It is generally known
that a mutation in the TRP sequence could cause drastic effect in the channel function. In the TRPV
channel, for instance, it has been reported that indole nitrogen in tryptophan makes strong hydrogen
bonds with carbonyl oxygen in the S4-S5 linker sequence. Mutation of tryptophan resulted in the
gain-of-function mutant of the TRPV3 (W692G) and TRPV4 (W773R) channels [57,58]. 2. General Architecture of TRPC4 and TRPC5 Channels The effect may
be partly attributable to structural proximity among the TRP helix, S4-S5 linker, and pre-S1 elbow. The next helical structure runs diagonally from the channel periphery towards the inner vestibule. This helical structure is called the ‘rib-helix’ or ‘connecting helix’ and is located distal to the TRP helix. The structure between the TRP helix and the connecting helix is missing in the Cryo-EM structure of
both TRPC4 and TRPC5. The connecting helix of one subunit penetrates the space between the ARD
of the subunit and ARD of the adjacent subunit (Figure 1). Hence it is generally conceived that the
connecting-helix may have an important role in inter-subunit interaction. After the connecting helix, a
longitudinal ‘coiled-coil helix’ or ‘coiled-coil domain’ follows. be found not only in TRPC channels but in the TRPM and TRPV superfamily. It is generally known
that a mutation in the TRP sequence could cause drastic effect in the channel function. In the TRPV
channel, for instance, it has been reported that indole nitrogen in tryptophan makes strong hydrogen
bonds with carbonyl oxygen in the S4-S5 linker sequence. Mutation of tryptophan resulted in the
gain-of-function mutant of the TRPV3 (W692G) and TRPV4 (W773R) channels [57,58]. The effect may
be partly attributable to structural proximity among the TRP helix, S4-S5 linker, and pre-S1 elbow. The next helical structure runs diagonally from the channel periphery towards the inner vestibule. This helical structure is called the ‘rib-helix’ or ‘connecting helix’ and is located distal to the TRP helix. The structure between the TRP helix and the connecting helix is missing in the Cryo-EM structure of
both TRPC4 and TRPC5. The connecting helix of one subunit penetrates the space between the ARD
of the subunit and ARD of the adjacent subunit (Figure 1). Hence it is generally conceived that the
connecting-helix may have an important role in inter-subunit interaction. After the connecting helix, a
longitudinal ‘coiled-coil helix’ or ‘coiled-coil domain’ follows. Based on topology, it is presumed that there are further cytoplasmic domains distal to the
coiled-coil helix, which in this article we call ‘far south domains (FSD)’. Moreover, many of the
key domains or amino acids responsible for channel tetramerization, G-protein mediated gating, or
membrane trafficking, are located in the FSD. We will discuss them in detail later in the article. 4 of 24 Cells 2020, 9, 73 Figure 1. General architecture of the TRPC4 and TRPC5 channels. 3.1. Conduction Pathway and Selectivity A pivotal study describing conduction pathway of the TRPC4 and TRPC5 channels was done in
embryonic brain and human embryonic kidney (HEK) 293 cells stably expressing human muscarinic
receptor subtype 1 (HM1 cell) [15]. When TRPC5 was over-expressed in the HM1 cell, they elicited large,
double-rectifying cationic current in response to extracellular 100 µM carbachol, a potent acetylcholine
analogue. However, mutant TRPC5 channel whose LFW moiety at 575–577th position was changed to
alanine (TRPC5LFW/AAA) failed to elicit any of cationic current in response to muscarinic stimulation. The mutant had no defect in trafficking to plasma membrane, hence suggested as a pore-collapsed
form of the channel. Moreover, co-expression of TRPC5LFW/AAA with wild-type channel severely
d
d
h l
ll
t
f TRPC4
d TRPC5 b t
t TRPC3
ti
th t th TRPC4
d The mutant had no defect in trafficking to plasma membrane, hence suggested as a pore-collapsed
form of the channel. Moreover, co-expression of TRPC5LFW/AAA with wild-type channel severely
reduced whole-cell current of TRPC4 and TRPC5 but not TRPC3, suggesting that the TRPC4 and
TRPC5 channels can share pore architecture in tetrameric assembly, whereas TRPC3 cannot. About 10 years later, similar results were observed [59]. In HEK293 cells expressing muscarinic
receptor subtype 3 (mAChR3), over-expression of both TRPC1 and TRPC4 yielded outward-rectifying
cationic current of heteromeric TRPC1/4 channel in response to 100 µM carbachol. When LFW moiety
of the TRPC1 channel was changed to alanine, the whole-cell current of the TRPC1/4 channel was
abolished. Although the pore may have been collapsed, the TRPC1LFW/AAA channel retained its capacity
for subunit assembly. Föster resonance energy transfer (FRET) signal from TRPC1LFW/AAA-CFP and
TRPC4-YFP was intact in plasma membrane. Cryo-EM structure of TRPC4 and TRPC5 locate the LFW moiety of both channels at pore-helix. At the pore helix, delocalized electrons of π system of aromatic rings of phenylalanine and tryptophan in
LFW moiety seem to have π–π interaction with each other and stabilize the entire pore structure [51,52]. The structures also show that from the selectivity filter to inner gate, arginine, glycine, isoleucine,
arginine, and glutamine may play significant roles in the conduction pathway. In TRPC5, mutations
targeted for those residues abolished current response to extracellular lanthanides (Gd3+). The mutants
showed diverse results in response to extracellular Englerin-A. N584A showed only a minute amount
of the current which is much smaller than wild-type. 3. Ion Conduction Pathway, Selectivity, and Rectification 3.1. Conduction Pathway and Selectivity 2. General Architecture of TRPC4 and TRPC5 Channels (A,B) Tetrameric structures of the
channels. Upper panel: surface map of the channels; Lower panel: cartoon diagram of the channels. (C) Monomers of each channel. The colors of each domain correspond to the domain diagram in
(D,F). (D) Domain diagram of TRPC4 and 5 channels. From the N-terminal cytoplasmic domain to
the C-terminal cytoplasmic domain, ARD, HLH, Pre-S1 elbow, Pre-S1 helix, VSLD, S5, S5-S6 linker,
pore helix, S6, TRP helix, connecting helix, and the coiled-coil domain are shown. (E) Relationship
between ARD and the connecting helix. Blue domains: ARD; Green domains: connecting-helix; Yellow
domains: coiled-coil domains. (F) Amino-acid numbers corresponding to each domain. Figure 1. General architecture of the TRPC4 and TRPC5 channels. (A,B) Tetrameric structures of the
channels. Upper panel: surface map of the channels; Lower panel: cartoon diagram of the channels. (C) Monomers of each channel. The colors of each domain correspond to the domain diagram in
(D,F). (D) Domain diagram of TRPC4 and 5 channels. From the N-terminal cytoplasmic domain to
the C-terminal cytoplasmic domain, ARD, HLH, Pre-S1 elbow, Pre-S1 helix, VSLD, S5, S5-S6 linker,
pore helix, S6, TRP helix, connecting helix, and the coiled-coil domain are shown. (E) Relationship
between ARD and the connecting helix. Blue domains: ARD; Green domains: connecting-helix; Yellow
domains: coiled-coil domains. (F) Amino-acid numbers corresponding to each domain. 5 of 24 Cells 2020, 9, 73 3.2. Rectification One of the most fascinating features of TRPC4 and TRPC5 channels is the shape of I-V curve [34]. In physiological membrane potential (−90 mV to 40 mV), the I-V curves of both channels mimic the
inward rectifier, such as inwardly-rectifying potassium channels. When the membrane potential is
depolarized more than 40 mV, both channels elicit sudden burst of outward current. This unique
feature of I-V curve is often called double-rectifying shape. Extremely low conductance in membrane
potential range of 0 mV to 40 mV is a characteristic feature of the channel and can be used as an
electrophysiological index to identify whether a measured current across the membrane is due to TRPC4,
5 channels or not. The low conductance at depolarized potential (0 mV to 40 mV) of TRPC4, 5 channels
is as important as an experimental index but so much as a physiological implication. It is well known
that TRPC4 or 5 channels show membrane excitatory effect. This may be due to their non-selectivity,
near-zero reversal potential and calcium permeability. But most of all, an inwardly-rectifying I-V
curve takes great part. TRPC4 or 5 channels, a mild membrane depolarizer, can efficiently clamp the
membrane potential at near zero because the channel permits no outward current at positive membrane
potential. Conversely, had TRPC4 or TRPC5 channels somewhat lost their inwardly-rectifying manner
in physiological potential range and become an outward-rectifier, their role would have been changed
to membrane stabilizers rather than depolarizers. This sudden change can be achieved whenever
TRPC1 is incorporated to TRPC4 or TRPC5 channels. In other words, heteromeric TRPC1/4 or TRPC1/5
channels show outwardly-rectifying I-V curves. An extensive review regarding electrophysiological
difference between homomeric and heteromeric channels can be found elsewhere [2,3,34,62]. One of the earliest studies demonstrating molecular mechanism of inward rectification of TRPC5
channel showed that intracellular magnesium ([Mg2+]i) blocks the channel at depolarized potential,
by binding aspartate residue D633 of the channel [63]. Mutation of D633 to asparagine abolished
inward-rectification feature of the channel in single channel conductance level. It is also worthy
to remark that mutation to D633N not only changed I-V shape but critically diminished absolute
amount of inward current through the channel. These results have suggested that the aspartate residue
is crucial for crucial for magnesium binding and inward-rectification, and the stabilization of the
residue by electrostatic interaction with Mg2+ seems to be necessary for the channel to elicit inward
cationic current. 3.1. Conduction Pathway and Selectivity G581A showed large current but fast inactivation
with near zero steady-state conductance was followed. Mutating three amino acids near the putative
inner gate (I621A/N625A/Q629A) elicited large current and slow inactivation was observed [51]
(Table 1). Table 1. Result of site-directed mutagenesis targeted for residues in ion conduction pathway. Residues
Gd3+
Englerin-A
N584A
-
+, minute activation
G581A
-
+++, fast inactivation
I621A/N625A/Q629A
-
+++, slow inactivation Table 1. Result of site-directed mutagenesis targeted for residues in ion conduction pathway. Since both Cryo-EM structures of TRPC4 and TRPC5 show closed state, it is still not clear how
conducting ions interact with amino acids within conduction pathway. Likewise, it is still hard to
explain which residues or cooperation of such residues are responsible for non-selectivity to cations,
namely, permeability ratios such as PCs/PNa or PCa/PNa. If we are to make a model based on conduction
pathways of already-published structures such as TRPV channels [60,61], TRPC may have two
independent gates governing conduction pathway. One may locate near the selectivity filter (upper
gate) and the other may locate near the cytoplasmic end of the conduction pathway (lower gate)
(Figure 2). 6 of 24 Cells 2020, 9, 73 Figure 2. Putative pore models of TRPC4 and 5 channels based on conformation changes shown in
TRPV2 channel. (A) Pore structure of TRPC4 (green) and TRPC5 (blue) channels. (B) Pore structure of
TRPV2 channels. Left panel: closed state; Middle panel: open state with widened lower, helix-bundle-cross
(HBC) gate; Right panel: open state with widened selectivity filter gate (SF gate) and HBC gate. Figure 2. Putative pore models of TRPC4 and 5 channels based on conformation changes shown in
TRPV2 channel. (A) Pore structure of TRPC4 (green) and TRPC5 (blue) channels. (B) Pore structure of
TRPV2 channels. Left panel: closed state; Middle panel: open state with widened lower, helix-bundle-cross
(HBC) gate; Right panel: open state with widened selectivity filter gate (SF gate) and HBC gate. 3.2. Rectification A similar mechanism was also observed in inwardly rectifying potassium channel
where [Mg2+]i [64–66] and intracellular polyamines [67–69] such as spermine and spermidine block
the cytoplasmic entrance of the pore and literally plug it at depolarized potential. The studies of
inwardly rectifying potassium channels have demonstrated that intracellular polyamines can make
stable electrostatic interaction with negatively charged amino acids within the pore since polyamines
are polycationic molecules with 2+~4+ valences in physiological pH [70]. Cells 2020, 9, 73 7 of 24 Interestingly, there had been studies suggesting that polyamines could modulate the activity of
TRPC4 channels. In guinea pig ileal myocytes, Tsvilovskyy et al. showed that cationic current evoked
by muscarinic stimulation (mIcat) was inhibited by extracellular polyamine, especially spermine [71]. The inhibition was not voltage-dependent and I-V curve retained doubly-rectifying shape. Moreover,
they also mentioned that concentration of extracellular polyamines in GI tract was in mM range and
intracellular concentration of polyamines was in µM level. Later, two pivotal studies showed that
molecular candidate for mIcat in ileal myocyte is TRPC4 and TRPC6 channels and that cholinergic
activation, namely, muscarinic receptor subtype 2 and 3 are governing the channel function in the
tissue [20,22]. [
]
The effect of intracellular polyamine to TRPC4 channel was verified later. With mutant TRPC4
channels, Kim et al. showed that glutamate residues at 728th and 729th position are possible binding sites
for intracellular spermine [72]. Intracellular spermine reduced outward current dramatically, leaving
inward current less changed. This asymmetrical current inhibition suggested voltage-dependency
of TRPC4 channel block by intracellular spermine. The claim was supported by the left shift of
conductance curve of TRPC4 channels. A different study with similar experimental setup claimed that
there exist additional binding sites for spermine [73]. In this study, the effects of D629A mutation have
been addressed in TRPC4 channels. D629 is equivalent site to D633 of TRPC5, hence may be responsible
for Mg2+ block and channel stabilization. Like TRPC5, D629A mutation reduced absolute current
of TRPC4 channel. Moreover, spermine could not block D629A channel as efficiently as wild-type,
suggesting that D629 may be involved in spermine binding. All residues deemed to be involved in
spermine-mediated inward-rectification are highlighted (Figure 3) based on Cryo-EM structure of
TRPC4 channel. Most of the residues found in functional studies circumference inner vestibule and
lower border of inner gate, except for the two: E648 and E649. 4.1. General Aspect of Homo- and Hetero-Tetramerization As described in the Introduction, TRPC1 can be found in almost every tissue in mammalians
whereas expression of TRPC4 or TRPC5 is rather specific. Given that the expression of each channel
subunits is tightly regulated, there exists nonzero chance that population of channels composed of
TRPC subunits is heterogeneous. In other words, all the results of possible combinations of subunit
assembly can be observed in a plasma membrane of a single cell. For instance, if a smooth muscle cell
in ileum expresses both TRPC1 and TRPC4, both the heteromeric TRPC1/4 channel and homomeric
TRPC4/4 channel could be found in plasma membrane. One of the earliest studies addressing the issue was done in HEK293 and Chinese hamster ovary
(CHO)-K1 cells using over-expression system of TRPC1, 3, 4, 5, and 6 channels [74]. Co-expression
of dominant-negative form of TRPC6, i.e., pore-collapsed form of TRPC6 (TRPC6LFW/AAA) abolished
TRPC3 current but not TRPC4 and TRPC5 current. The study also designed fluorescence protein tagged
channels of which CFP or YFP tags are linked at C-terminus of the channel. Using such constructs, the
study evaluated which combination is most feasible. As a result, TRPC1, 4, and 5 showed strong FRET
signal with one another. TRPC3 and 6 also showed high FRET signal. In contrast, no FRET signal was
observed between two groups. Co-immunoprecipitation (Co-IP) yielded similar results. As mentioned
earlier, heteromerization between TRPC1, 4, and 5 was also observed in embryonic brain [15]. A recent
study demonstrating neuro-protective effect of TRPC1 also showed that both TRPC1 and TRPC5 are
expressed in substantia nigra and heteromeric population (TRPC1/5) is substantial in normal tissue [13]. They have argued that excessive Ca2+ influx through homomeric TRPC5 channel is the key pathologic
step in Huntington’s disease and the pathologic Ca2+ influx is triggered by reduced expression of
TRPC1, hence increased portion of homomeric TRPC5 in plasma membrane. Based on nearly 20 years of cumulative results, it is now well acknowledged that all TRPC1, 4,
and 5 subunits contribute equally to form a conducting pore in a tetrameric assembly. In a sense,
the difference in major channel characteristics such as activation mechanism, selective permeability,
pharmacological sensitivity, and membrane excitatory effect among TRPC1/4, TRPC1/5, TRPC4/4, and
TRPC5/5 channels could not be ignored. Truly, the strength of electric field exerted by amino acids in
selectivity filter was starkly different between homomeric TRPC4/4 channels and heteromeric TRPC1/4
channels. 3.2. Rectification These residues locate right after TRP
helix and have spatial proximity to pre-S1 elbow. These residues may attract cytosolic spermine and
increase the local concentration of spermine near the channel periphery. Overall effect of E648 and
E649 would be reducing access resistance for the following reaction: [Spermine]cytosol →[Spermine]inner vestibule (1) Figure 3. (Red sticks) Residues responsible for Mg2+ and spermine-mediated inward-rectification of
TRPC4. Distances in Angstrom between D629 residues (18.1) and E728 residues (12.5) are shown. (Blue sticks) Residues responsible for putative inner gate of the channel: I617, N621, and Q625. Figure 3. (Red sticks) Residues responsible for Mg2+ and spermine-mediated inward-rectification of
TRPC4. Distances in Angstrom between D629 residues (18.1) and E728 residues (12.5) are shown. (Blue sticks) Residues responsible for putative inner gate of the channel: I617, N621, and Q625. In a sense, it seems quite surprising that intracellular spermine cannot affect heteromeric
channels, i.e., TRPC1/4 and TRPC1/5 channels [72]. Since the I-V curve of heteromeric channels
is outwardly-rectifying, intracellular spermine may strongly reduce the outward current if they can
exert similar cytoplasmic blocking effect to the heteromeric channel. The stark difference between Cells 2020, 9, 73 8 of 24 homomeric and heteromeric channel may be attributed to different structure of two channels, especially
c-terminal domain and inner vestibule. In summary, the studies of TRPC5 and TRPC4 emphasize that amino acids constituting inner
vestibule are crucial for inward-rectification of the channel. Intracellular magnesium bind to a aspartate
residue which exist in close proximity with one another in tetrameric formation and the attractive
electrostatic interaction between magnesium and aspartate is necessary to elicit stable cationic current
through the channel. Acidic amino acids in 720–740 region of TRPC4 channels are crucial for
spermine-mediated inward-rectification. 4.1. General Aspect of Homo- and Hetero-Tetramerization Namely, the Eisenmann sequence, an estimate for pore field strength, was different between
two channels. Homomeric channels showed weak pore-field strength while heteromeric channel
showed strong pore-field strength [59,75]. 4.2. Molecular Mechanism of Tetramerization Process The studies above are milestone of the endeavor for understanding the tetramerization process of
TRPC channels. Information given by the studies such as the existence of heteromeric TRPC channels
in plasma membrane and an empirical proof for combinatorial number of cases all helped to deepen
our understanding of the topic. Despite of those elegant studies, however, the molecular mechanism
of tetramerization was still unknown. Cells 2020, 9, 73 9 of 24 In the late 2000s, molecular mechanisms of tetramerization among TRPC4 and TRPC5 channels
were addressed from an Austrian group and a Canadian group. Using FRET method and size-exclusion
chromatography, they suggested that a part of 1st ARD (69–98) is crucial for homotetramerization
process of TRPC5 [76], and that parts of 3rd and 4th ARD (87–172) and N-terminus coiled-coil domain
(254–304) are important in the homotetramerization process of TRPC4 [77]. Their arguments were
based on the channel topology at the time. Up-to-date domains are 2nd ARD (69–98) of TRPC5, 2nd and
3rd ARD (87–172), and helix-loop-helix (HLH) domain (254–304) of TRPC4. Based on domain-specific
results, one study postulated two possible models for the homotetramerization process of TRPC4
channels. The first model assumed that each subunit has full capacity for tetramerization and no
further constraints were necessary. The second model assumed that two monomers should form dimer
first, generating two dimers in total. This initial dimerization process is achieved using 3rd ARD
(87–172) according to the model. Then, two dimers are assembled together to form a tetramer by
interactions of HLH domains (254–304). A recent study suggested similar results using FRET method
and thorough electrophysiology. With a number of N-terminal truncation mutants of TRPC4 channels,
the study argued that 98–124 residues in N-terminus (3rd ARD) and 700–728 residues in the C-terminus
(connecting-helix) play a great role in the homotetramerization process of TRPC4 [78]. In theory, the large quaternary structure of TRPC channels would require delicate post-translational
modification in various intracellular organelles, especially in terms of tetrameric assembly. Therefore,
finding out the time and the location of the tetramerization process would be both intriguing and
necessary. Recently, there has been a report that tetramerization process may occur in endoplasmic
reticulum (ER)-Golgi system [79]. When 553rd cysteine residue of TRPC5 was mutated to serine or
alanine, a membrane translocation was severely damaged. Expression level of TRPC5 channel at
plasma membrane was significantly low in TRPC5C553S mutant compared to wild-type (WT) channel. 4.2. Molecular Mechanism of Tetramerization Process Moreover, co-expression of TRPC5C553S mutant even reduced surface expression level of wild-type
channel. The most interesting part of the study is that high FRET signal was detected inside Golgi
complex when TRPC5C553S-CFP and TRPC5WT-YFP were co-expressed. Based on the results, the study
claimed that C553 is crucial for membrane translocation process of TRPC5 and the tetramerization
process may occur at or before Golgi. The molecular mechanistic study for heteromerization process is unfortunately limited despite of
its significance. One study so far represented specific domains responsible for heteromerization process
of TRPC1/4 and TRPC1/5 channels. Using FRET method and electrophysiological recording subjected
to various truncation mutants, Myeong et al. suggested that 700–728 residue (connecting-helix) of
TRPC4 and 707–735 residue (connecting-helix) of TRPC5 are important in heteromerization with
TRPC1 [80]. It is surprising, however, that TRPC1 utilizes different domains for TRPC4 and TRPC5. For TRPC1/4, 725–745 region of TRPC1 is used as an inter-subunit interface, while the 673–725 region
is used for TRPC1/5. Topologic analysis based on sequence alignment and Cryo-EM structure of
TRPC4 and TRPC5 suggests that both regions of TRPC1 correspond to putative connecting-helix of
TRPC1 channel. In summary, both N-terminal and C-terminal cytoplasmic domains seem to have crucial role in
tetramerization process among TRPC1, 4, and 5 channels. As mentioned earlier in general architecture,
ARDs and connecting-helices are of special interest. Last but not least, cysteine residue in the S5-S6
linker seems to have a great role in tetramerization process (Figure 4, Tables 2 and 3). Indeed, two
cysteines in S5-S6 linker, C549/C554 in TRPC4 and C553/C558 in TRPC5, bear many complicated
functions of which we will discuss in the following section. 10 of 24 Cells 2020, 9, 73 Table 2. Summary of domains responsible for homo- or hetero-tetramerization of TRPC 4 channels. y
p
Region (Residue)
Reference
1
2nd and 3rd ARD (87–172)
[77]
2
3rd ARD (98–124)
[78]
3
HLH (254–304)
[77]
4
Connecting Helix (700–728)
[78,80] Table 3. Summary of domains responsible for homo- or heterotetramerization of TRPC 5 channels. Region (Residue)
Reference
1
2nd ARD (69–98)
[76]
2
Connecting Helix (707–735)
[80]
3
C553
[79] Figure 4. Domains responsible for tetramerization process in TRPC4 and TRPC5 channels. (A) Domains
in TRPC4 channels. Blue domains: ARD; Light green domains: HLH; Green domains: connecting helix. (B) Domains in TRPC5 channels. Blue domains: ARD; Green domains: connecting-helix. Figure 4. Domains responsible for tetramerization process in TRPC4 and TRPC5 channels. 4.2. Molecular Mechanism of Tetramerization Process (A) Domains
in TRPC4 channels. Blue domains: ARD; Light green domains: HLH; Green domains: connecting helix. (B) Domains in TRPC5 channels. Blue domains: ARD; Green domains: connecting-helix. 5.1. Functional Studies of Cysteine Modificaiton Two years later, another study reported that
reduction of cysteine residues by extracellular reducing agent such as dithiothreitol (DTT) activates
the TRPC5 channel and that C553 and C558 are target sites for reduction reaction [84]. According to
the study, TRPC5 gains activity when C553 or C558 has free thiol group rather than oxidized form. Another comprehensive study for C553 and C558 came out about 10 years later. In this article, Hong et al. showed that C553S, C558S, and C553S/C558S mutants are all irresponsive to muscarinic stimulation,
extracellular lanthanides, and reducing agent (DTT) [79]. The study suggested that cysteine residues
are involved in tetramerization process and membrane translocation process of TRPC5 channels. In view of whole-cell current of such mutants, findings suggested by one of the latest studies in the
topic looks surprising. Jeong et al. showed that C553S and C558S mutant TRPC5 channels could not
elicit cationic current in response to extracellular Englerin-A [85]. However, double mutant of both
cysteines (C553S/C558S) elicited large Englerin-A-mediated current. As we have seen through, accumulated results of C553 and C558 show some inconsistencies
among one another. The current response to reducing agent, muscarinic stimulation, lanthanides,
Englerin-A and plasma membrane expression level of mutant channels are different from one study to
another. We summarized the key findings regarding such issues in Table 4, from a number of reports,
which we expect to be helpful to gain functional insight. Table 4. Summary for accumulated results investigating outcome of cysteine modification. Table 4. Summary for accumulated results investigating outcome of cysteine modification. 5.1. Functional Studies of Cysteine Modificaiton The thiol group (-SH) of cysteine grants chemical flexibility to the protein. Free thiol group can
undergo numerous chemical reactions and the reaction pathway varies according to the type of reactant
partner, redox potential of reaction compartment, and many others. One of the most studied reaction
of thiol group in a biological system would be formation and dissociation of disulfide bond between
two cysteine residues. It is generally accepted that the first formation of disulfide bridge of a protein
occurs inside ER [81,82]. Since both intracellular and extracellular fluid can contain diverse enzymes
or biomolecules for rigorous redox reaction, however, a preformed disulfide bridge can be broken at Cells 2020, 9, 73 11 of 24 certain condition. Likewise, free thiol groups can be oxidized by cellular oxidants such as glutathione
and form disulfide bridge outside ER. g
Among number of cysteine residues in TRPC channels, two cysteines (C549/C554 of TRPC4
and C553/C558 of TRPC5) have been studied most intensively. The cysteine residues are conserved
in TRPC1, 4, and 5 channels but not in TRPC3 and 6 channels [83]. The earliest study was about
S-nitrosylation of cysteine residues of TRPC5 channel and the outcome of such reaction onto the
channel activity [83]. Yoshida et al. showed that increasing NO concentration using NO-donating
chemical (S-nitroso-N-acetyl-dl-penicillamine, SNAP) induced robust TRPC5 activity [83]. The effect
was independent from cGMP pathways suggesting that increased cytosolic NO may directly interact
with channel protein. A similar effect was observed when a chemical, which can undergo S-nitrosylation
reaction with thiol group (5-nitro-2-PDS), was applied. Using cell-impermeable chemicals, they also
suggested that such S-nitrosylation reaction may occur through a route from cytosolic space to the free
thiol residue of the channel. In conclusion, the study claimed that S-nitrosylation at C553 could result in
the opening of the channel via cytosolic approach and that channel closes when intracellular reducing
agent reduces cysteine residue to –SH. Along with the results above, the study also showed that
despite of their irresponsiveness to S-nitrosylation C553S and C558S can be activated by conventional
activation mechanisms such as muscarinic stimulation. Moreover, the surface level of C553S and
C558S mutant was indifferent from wild-type TRPC5 channel, suggesting that two cysteine residues
rarely affect membrane translocation of the channel. 5.2. Structure–Function Relationship in Cysteine Modification 5.2. Structure–Function Relationship in Cysteine Modification 5.2. Structure–Function Relationship in Cysteine Modification 5.2. Structure–Function Relationship in Cysteine Modification Cryo-EM structure of mouse TRPC5 channel revealed that both C553 and C558 are located in
S5-S6 linker. Likewise, C549 and C554 are also located in S5-S6 linker of TRPC4 channels. In both
structural studies, authors not only revealed elegant Cryo-EM structures of the channels but also
presented some critical electrophysiological recordings [51,52]. In TRPC4, extracellular DTT could
not activate C549A and C554A mutant channels. The result would be probably due to the absence of
active cysteines. When extracellular Englerin-A was applied, both C549A and C554A could not elicit
current in response to the drug. Surprisingly, however, C549A/C554A double mutant elicited strong
cationic current in response to Englerin-A. This result is somewhat in line with previously-mentioned
C553S/C558S case [85]. In TRPC5, C553S, C558S, and C553S/C558S all failed to show significant current
response to DTT or Englerin-A [51]. Based on the aforementioned structure and electrophysiological
recording of TRPC4 channels, Duan et al. examined whether there exists any difference in S5-S6 linker
sequence around two cysteine residues of TRPC4 and TRPC5 channels [51]. As a result, they have
found that both channels share exactly same amino acid sequence between two cysteine residues,
namely, ‘CKGIRC’. However, sequences upstream to first cysteine were different. TRPC4 has ‘ETKGLS’
sequence, while TRPC5 has ‘TRAIDEPNN’ sequence. Based on this speculation, they made chimeric
TRPC5 channel whose S5-S6 linker sequence ‘TRAIDEPNN’ is changed to the one of TRPC4 (‘ETKGLS’)
and examined how this chimeric TRPC5 channel respond to Englerin-A. In short, changing S5-S6 linker
sequence could not make chimeric TRPC5 channel responsive to Englerin-A. Structure gives clear location of key cysteine residues, namely, C549 and C554 in TRPC4 channels,
and C553 and C558 in TRPC5 channels. Moreover, their location—S5-S6 linker—gives good explanation
for their accumulated functional significance. Functional implication, however, looks way more
entangled. It may be due to the cruel fact that cysteine modification is harnessed with various cellular
mechanisms such as redox potential, tetramerization, ER-Golgi-plasma membrane trafficking process
and protein folding. 5.1. Functional Studies of Cysteine Modificaiton Current Response to
Surface Level
La3+ or Gd3+
Englerin-A
Reducing Agent
Muscarinic Stimulation
TRPC5
C553X
Reduced [51]
Reduced [79]
Reduced [51]
Reduced [51]
Reduced [79]
Reduced [79]
Reduced * [83]
Reduced [51]
Reduced [79]
Intact [83]
C558X
Reduced [51]
Reduced [79]
Reduced [51]
Reduced [51]
Reduced [79]
Reduced [79]
Reduced * [83]
Reduced [51]
Reduced † [79]
Intact [83]
C553X/C558X
Reduced [51]
Reduced [79]
Reduced [51]
Intact [85]
Reduced [51]
Reduced [79]
Reduced [79]
Reduced * [83]
Reduced ‡ [85]
Reduced [51]
Reduced [79]
Intact [83]
TRPC4
C549X
N/A
Reduced [53]
Reduced [53]
N/A
N/A
C554X
N/A
Reduced [53]
Reduced [53]
N/A
N/A
C549X/C554X
N/A
Intact [53]
Reduced [53]
N/A
N/A
N/A: Not addressed, X: alanine or serine; * Channel activity measured by Ca2+ indicator and F340/F380 but not
current; † Authors specifically mentioned in the article that only minute amount of surface level was reduced; ‡
overexpression of gain-of-function mutant of Gαi2. Cells 2020, 9, 73 12 of 24 12 of 24 6.1. General Aspect of G-Protein Mediated Gating Mechanism As mentioned earlier in the article, G-protein coupled receptors (GPCRs), heterotrimeric G-proteins,
and G-protein signal transduction pathways are one of the most classic activation mechanisms of
TRPC channels. Among three most prominent subtypes of heterotrimeric G-proteins, namely, Gs, Gi
and Gq, it was Gq-protein pathway and its implication to channel activation mechanism that have
been studied most intensively. However, dissecting Gq pathway alone has been challenging. First,
it is partly due to inherent complexity and dynamics within signal transduction pathway; one must
consider all biomolecules participating in the signal transduction pathway—Gαq, Gβγ, PLC-β, PIP2,
inositol 1,4,5-triphsophate (IP3), DAG, and PKC. Second, participants of signal transduction pathway
are in delicate chemical equilibrium, hence up- or down-regulating one component immediately affects
other components. For example, over-expressing gain-of-function mutant of Gαq may be useful
to attest the effect of Gαq to TRPC channels. The interpretation becomes cumbersome, however,
if at least one confounder participates in. For instance, if a finite amount of PIP2 was necessary
to maintain activity of the channels, long-term effect of over-expressed Gαq would turn out to be
negative to the channel activity regardless of its true nature. However, most of all, it was rapid and
prominent desensitization of the channels that made dissecting downstream components out of overall
effect of the Gq pathway to TRPC channels challenging. With an astonishing consistency, TRPC4 or
5 channels showed prominent desensitization right after strong activation whenever the channels were
activated by Gq-protein coupled receptors (GqPCR, e.g., mAChR1, or 3, histamine receptor subtype
1) [19,21,22,37–43,59,71,72]. As an alternative, a non-specific G-protein activator such as [GTPγS]i has
been used [35,45,46]. Although GTPγS-induced channel current showed less or no desensitization,
non-specificity of the drug still made the discrimination troublesome. Cells 2020, 9, 73 13 of 24 The development of Englerin-A made it possible to record G-protein independent, sustained
TRPC4 and 5 currents [48]. Moreover, a recent advancement in biochemical tools must be also
appreciated such as Gq proteins without PLC-β stimulatory effect, rapamycin-inducible systems, and
voltage-sensitive phosphatases. Those methods all contributed to discriminate components of Gq
downstream pathway and to measure individual effects to TRPC channels. 6.2. Diacylglycerol (DAG) and Protein Kinase C (PKC) Earlier studies showed that only TRPC3 and TRPC6 are sensitive to DAG and can be directly
activated by DAG or OAG [44]. Recent studies, however, showed that TRPC4 and 5 channels also have
sensitivities to DAG and can be activated by it [86,87]. The pre-requisite must be sufficed, however,
if TRPC4, 5 channels are to be activated by DAG. The channels elicited current in response to DAG
only when phosphorylation of threonine at C-terminus (T972 of TRPC5) by PKC had been prevented. This claim was supported by the result that OAG elicited TRPC5 current only when PKC inhibitor
was pretreated. TRPC5T972A mutant channel showed cationic current in response to OAG. The study
also correlated PKC phosphorylation to the PDZ-domain of TRPC4 and 5 channels. The existence of
PDZ-domain (VTTRL) in C-terminus of TRPC4, 5 channels and their interaction with NHERF (Na+/H+
exchanger regulatory factor) protein have been already known. Since NHERF protein could serve as a
scaffolding protein between cytoskeleton and any given protein containing PDZ-domain within, the
verification of interaction per se between PDZ-domain containing TRPC channels and NHERF held
much significance. The functional relevance for such interaction, however, was scarce. Using shRNA
to NHERF proteins, the study has shown that TRPC5 current can be elicited by OAG when NHERF
was down-regulated by shRNA. The involvement of PKC in functional regulation of TRPC5 was also raised in another study but
with different context [43]. What the study has found is that desensitization of carbachol-activated
TRPC5 current was slowed as intracellular calcium increased. Thus, it was supposed that desensitization
process may be calcium-dependent. They also have shown that desensitization process was slowed
down when PKC inhibitors were treated. TRPC5T972A mutation also yielded full activation but no
desensitization under carbachol stimulation. The study concluded that desensitization process shown
in GqPCR activated channels is partly due to PKC phosphorylation of the channel at T972 residue. 6.3. Phosphatidylinositol 4,5-bisphosphate (PIP2) For instance, A-FKBP and FRB-B can
form a dimer (A-FKBP:rapamycin:FRB-B) once the rapamycin is supplied in the system. The binding
of rapamycin onto FKBP or FRB is noncovalent, hence reversible. Like in TRPC5, carbachol (mAChR3) activated TRPC4β current measured in HEK293 cells showed
strong activation followed by desensitization [39]. When PIP2 was added in intracellular solution, no
desensitization of TRPC4β was observed. The study designed Lyn11-FRB and FKBP-Inp54p system
where Lyn11 is a membrane-bound protein and Inp54p is an enzyme that can hydrolyze PIP2 into
PI(4)P. In such a system, rapamycin would recruit Inp54p to the inner border of plasma membrane,
which unless otherwise would be located diffusely in the cytosol. Unlike muscarinic stimulation,
GTPγS-activated TRPC4β current shows slow activation but no prominent desensitization. What the
study has found is that for Lyn11-FRB and FKBP-Inp54p expressing cell, a sudden desensitization
process was observable in GTPγS-activated TRPC4β current as soon as rapamycin was applied. Moreover, the study also showed that the desensitization process of TRPC4β is independent from
phosphorylation of the channel by PKC since the desensitization of the channel was still observable
even in the presence of PKC inhibitor (chelerythrine, GF109203X). The desensitization was also
retained in TRPC4βT887A channel where T887 is equivalent site for T972 in TRPC5, hence putative PKC
phosphorylation site of TRPC4β channel. A different study was reported which shares the similar context that PIP2 is necessary to maintain
the opening of TRPC4β [37]. In this study a voltage-sensitive phosphatase (VSP) system was employed. VSP has an ability to hydrolyze PIP2 into PI(4)P and phosphate and the enzyme performance is
increased once depolarized pulse is applied. The more positive and more frequent the pulse is, the
more enzymatic reaction of PIP2 hydrolysis by VSP can be. Using the VSP system, the study has shown
that the Englerin-A-activated current of TRPC4α, TRPC4β and TRPC5 channels were all inhibited by
activation of VSP by depolarized pulse sequence. The study also monitored PIP2 level and whole-cell
current simultaneously, calculating PIP2 sensitivity (IC50 value for I/Imax) of each channel. PIP2 level
was monitored by FRET signal coming from PH-CFP and PH-YFP. PH domain is a subdomain of PLC-δ
responsible for high affinity of PLC-δ to PIP2. The calculated Kd values for PIP2 showed following
relationship: KdC4β < KdC4α < KdC5. 6.3. Phosphatidylinositol 4,5-bisphosphate (PIP2) There are a number of issues surrounding PIP2 and activity of TRPC1, 4, and 5 channels. One of
the earliest functional implications of PIP2 was addressed in the study that investigated alternative
splicing variants of TRPC4 channels. Both human and mouse TRPC4 channels, like other proteins, do
have natural splicing variants. One variant holds full exon sequence, hence called unspliced isoform or
TRPC4α. The other variant has 84 less amino acids than the former and is called TRPC4β. Alternative
splicing occurs at the C-terminal cytoplasmic domain of the channel (785–868th a.a. with respect to
TRPC4α). Using over-expression system of two splicing variants in HEK293 cells, a study suggested
that PIP2 exerts tonic suppressive effect to TRPC4α [42]. When diC-8, a water soluble analogue of PIP2
was added in intracellular solution, the GTPγS-induced whole-cell current of TRPC4α was reduced. On the other hand, the effect of PIP2 onto TRPC4β was minimal. Therefore, the authors concluded that
785–868 residue of TRPC4α is the discriminant for PIP2 governance to TRPC4 channels and that PIP2
may directly bind to the residue. Other phosphoinositides such as PI(3)P or PI(4)P showed no effect to
TRPC4 channel, hence the effect can be seen as PIP2-specific. Despite of tonic suppressive effect of
PIP2, PIP2 depletion alone could not activate both TRPC4 variants suggesting that the action of PIP2
has much meaning once the channel is opened by other activators. Other studies claimed that PIP2 is involved in desensitization process of TRPC5 channels [41]. In HEK293 cells, the whole-cell current of over-expressed mouse TRPC5 channels was elicited by
muscarinic receptor stimulation (mAChR3). Remark that stimulation of GqPCR strongly activates
TRPC channel but prominent desensitization follows through the activation. In normal conditions, Cells 2020, 9, 73 14 of 24 14 of 24 a mAChR3-activated TRPC5 current showed a similar time-course. When diC-8 was added in
intracellular solution, however, desensitization was significantly slowed. The result implicated that
desensitization can be partly attributed to decrease in PIP2. More direct investigation to desensitization process and PIP2 level was achieved using
rapamycin-inducible system. Rapamycin-inducible system utilizes rapamycin-dependent dimerization
process of two proteins: FKBP (FK506-binding protein) and FRB (FKBP-rapamycin-binding domain). Namely, for two proteins A and B that can be fused with FKBP or FRB, spatial proximity between A
and B can be achieved by making dimers out of FKBP and FRB. 6.3. Phosphatidylinositol 4,5-bisphosphate (PIP2) The study postulated two putative binding pockets for PIP2
within TRPC4β channel as well based on a calculated structural model of TRPC4β using the Cryo-EM
structures of TRPM4 and NOMPC channel at the time as the templates. K419, K664 constituted one
binding pocket and R551, K518, H630 constituted the other pocket (Figure 5). g p
p
g
It has been also addressed whether PIP2 exerts regulatory effect to heteromeric TRPC channels. Using VSP and rapamycin-inducible system, one study showed that desensitization of TRPC1/4β and
1/5 channels shown in carbachol stimulation is due to PIP2 depletion [38]. The paper which measured
Kd value for PIP2 inhibition also showed that desensitization of TRPC1/4α, TRPC1/4β, and TRPC1/5
are due to hydrolysis of PIP2. Moreover, they have shown that dispersed Kd values of TRPC4α, 4β and
5 channels converge to a singular value once TRPC1 is incorporated to the channels. In other words,
Kd values of TRPC1/4α, TRPC1/4β, and TRPC1/5 were not different from one another. A criticism was raised against the notion that desensitization process is solely dependent on
PIP2. One study claimed that there exist additional pathways that must be considered to explain
desensitization process of heteromeric TRPC1/4β and TRPC1/5 channels [88]. The study examined
carbachol-activated TRPC1/4β, TRPC1/5 current in mAChR3-expressing HEK293 cells with and
without PKC inhibitor. The desensitization was slowed down in TRPC1/5 when PKC inhibitor was Cells 2020, 9, 73 15 of 24 pre-incubated but not in TRPC1/4β. They also examined carbachol-activated current of TRPC1/4bT887A
and TRPC1/5T972A. The desensitization of TRPC1/4βT887A was not different from TRPC1/4β but
TRPC1/5T972A showed slower desensitization than TRPC1/5. In summary, it seems clear that PIP2, a plasma membrane constituent, yields a substantial effect to
the activity of TRPC channels. The alternative splicing of TRPC4 resulted in different sensitivity to
PIP2 modulation. Accumulated results suggest that for a sustained opening of the channel, a finite
amount of PIP2 near the channel is necessary. If the PIP2 level is decreased, activated channel seem to
fall into inactivation. PIP2 alone, however, could not activate the channel, suggesting that the effect
of PIP2 is meaningful only if the channel is opened by another activator. For this finding, the help
of direct channel activator, Englerin-A, was paramount. If we apply the context to GqPCR-activated
channels, activation of the channel must have been occurred before PIP2 hydrolysis begin to inactivate
the channel. 6.3. Phosphatidylinositol 4,5-bisphosphate (PIP2) The Gq signal transduction pathway leaves no other choices for activation mechanism
which is upstream to PIP2 hydrolysis: Gαq protein itself. Figure 5. Putative binding pocket for PIP2 in TRPC4 channels. (A) Two binding pockets shown in one
subunit of the channel. (B) First binding pocket consists of K664 and K419. (C) Second binding pocket
consists of R551, K518, and H630. Figure 5. Putative binding pocket for PIP2 in TRPC4 channels. (A) Two binding pockets shown in one
subunit of the channel. (B) First binding pocket consists of K664 and K419. (C) Second binding pocket
consists of R551, K518, and H630. 6.4. Direct Activation by Gαq Protein Early studies questioning the possibility of direct TRPC channel activation by Gαq protein showed
negative results [45,46]. It may be due to the lack of direct pharmacological agonist or antagonist
for Gαq protein. The alternative experimental setup was to heterologously express gain-of-function
mutant of Gαq proteins. Based on autonomic GTPase activity of alpha subunit of heterotrimeric
G-proteins, a gain-of-function mutant of Gαq protein was generated. Once the glutamine residue
crucial for autonomic GTPase activity is mutated to leucine, GTP bound form of alpha subunit, i.e.,
active Gαq could not hydrolyze the GTP, hence remain constitutively active. This Q to L mutation
would surely augment the activity of Gαq protein but might be too strong to see adequate activation
process of TRPC channels. In other words, even if Gαq could directly activate the TRPC channels,
the overall effect of over-expression of Gαq QL protein would turn out negative since all of the PIP2
necessary for maintaining the TRPC activity would have been depleted. The great breakthrough came out with the help of GαqQL/LA mutant. This mutant Gαq protein
has constitutive activity due to QL mutation but cannot interact with PLC-β, a downstream effector
molecule. In rapamycin-inducible system using Lyn11-FRB and FKBP- GαqQL, GTPγS-activated
TRPC4β current was gradually reduced right after rapamycin was introduced [39]. In Lyn11-FRB and
FKBP-GαqQL/LA, however, no change was observable. A similar system was conveyed to attest direct 16 of 24 16 of 24 Cells 2020, 9, 73 effect of Gαq protein onto TRPC1/4β and TRPC1/5 heteromers [38]. In short, rapamycin-induced GαqQL
could not activate TRPC1/4β or TRPC1/5 channels but GαqQL/LA could. Moreover, the interaction
between Gαq and TRPC channels was verified by Co-IP. The reason rapamycin-induced GαqQL could
not activate the channels is that the positive effect of GαqQL to the channel—direct activation—and
negative effect—PIP2 hydrolysis—occurs in a similar time-scale. This may be attributed to high
a turn-over rate of PLC-β or low initial-value of PIP2 at least in micro-domain level. How fast
rapamycin-induced dimerization of FKBP and FBR occurs would be also an important factor as well. Interestingly, multiplying the time-course of both PIP2 level in plasma membrane measured by YFP
signal from PH-YFP and level of GαqQL docked to plasma membrane measured by RFP signal from
GαqQL-RFP yielded highly similar curve to time-course of whole-cell current. 6.5. Direct Activation by Gαi Protein The very notion that G-proteins other than Gq-PLC pathway might activate TRPC channels was
hard to accept at the time probably because there had been enormous studies supporting the latter
pathway. The idea started from an astounding response of TRPC4 and 5 channels to pertussis toxin
(PTX). PTX is one of the most potent and specific blockers of alpha subunit of inhibitory G-proteins
(Gαi). Jeon et al. has found that both TRPC4β and TRPC5 channels elicit large currents when muscarinic
acetylcholine receptor subtype 2 (mAChR2) was stimulated by carbachol [46]. When PTX was treated,
however, mAChR2 could not elicit any current at all. They also have shown that gain-of-function
mutant of Gαi proteins (GαiQL) increases whole-cell current of both channels. This effect was also
abolished by PTX. Later, a more comprehensive study further extended the idea by showing that Gαi2
and Gαi3 are potent, direct activators of TRPC4β and TRPC5 channels, respectively [45]. The study
ruled out the involvement of beta-gamma subunits in Gαi-mediated activation process of the channels. Finally, the study suggested that K715 and R716 residue of TRPC4β are responsible for direct interaction
between Gαi protein. The pathophysiological implication of such activation mechanism was studied
in cultured hippocampal neurons [75]. Over-expression of Gαi2QL and TRPC4β channel resulted
in reduced neurite outgrowth of the neurons. The study claimed that this pathologic response is
due to excessive calcium influx via highly activated TRPC4β channels and subsequent activation of
calcium-calmodulin dependent protein kinase II (CaMKII) downstream pathways. Although Gαi proteins strongly activated TRPC4β channels with direct interaction, they couldn’t
activate TRPC1/4β heteromeric channels [59]. In HEK293 cells expressing both TRPC1 and TRPC4β
channels, a large outwardly-rectifying (TRPC1/4β) current was elicited when mAChR3 stimulation
was employed. On the other hand, only a minute amount of doubly-rectifying current (TRPC4β)
was observable when mAChR2 was stimulated. Over-expression of Gαi2 protein also elicited only
doubly-rectifying whole-cell currents. The difference in sensitivity to muscarinic governance (mAChR2
or mAChR3) between TRPC4/4 and TRPC1/4 channels may explain TRPC4/4-dominance in ileal
myocyte where mAChR2 governance was shown to be more dominant than mAChR3 [20,22]. 6.6. Structure–Function Relationship in G-Protein Mediated Gating Process First, DAG-mediated regulation requires a corresponding lipid-binding pocket in the channel
structure. For example, TRPV and TRPML, which are all activated by lipids—capsaicin and
PI(3,5)P2—show binding pockets for their agonistic lipids [89,90]. Although Cryo-EM structures show
some lipid binding pockets in both TRPC4 and TRPC5, conformation rejects the likelihood of the
pocket being DAG-specific. Even the structure of TRPC3 could not represent DAG-binding pockets
despite long-history for DAG-mediated gating mechanism of the channel [55,56]. One study, however,
proposed a promising result [91]. Using a modeled structure of TRPC3 with TRPV1 as a template,
putative DAG binding domain (G652) was postulated. Since sequence homology around the residue is
high among TRPC channels, we have marked identical glycine residue in TRPC4 channels (Figure 6). Second, two residues bear special importance in terms of PKC-mediated regulation of the channels: Cells 2020, 9, 73 17 of 24 T887 and T972. These residues are located in FSD and missing in Cryo-EM structure of both TRPC4
and TRPC5. 7 and T972. These residues are located in FSD and missing in Cryo-EM structure of both TRPC4
TRPC5. PIP2 holds exceptional position in Gq-mediated gating mechanism. In addition to functional
implication in channel gating mechanism, a physiological relevance is also immense. By the virtue of
calcium influx and membrane excitation, it carries substantial pathophysiological meaning whether
the channel opens in episodic manner due to inherent desensitization or the opened channel sustains
the opening or the channel stays refractory even in the presence of activators. In spite of studies mentioned above, domain-specific functional studies against PIP2-mediated
regulation of TRPC channels are still limited. K419, K664, and R511, K518, H630 are putative
binding sites at least with functional evidence. The difference in PIP2 selectivity between homomeric
channels and heteromeric channels may indicate that binding site for PIP2 might be overlapped
with subunit-subunit interface. Provided that heteromeric subunits locate at cis-position one another
rather than trans-position, PIP2 binding may require cooperative action of different residues in
cis-positioned subunits. In addition to the report that TRPC4α receives tonic suppression by PIP2 but not TRPC4β, a
number of studies also presented results supporting that there exists a difference in characteristics
between two splicing variants. Thus, a structural explanation for such event seems imperative. Unfortunately, however, the 84 amino acids are also included in FSD, hence no structural evidence can
be provided for now. 6.6. Structure–Function Relationship in G-Protein Mediated Gating Process In our best knowledge, no study has been addressed Gαq-binding site within TRPC1, 4, 5 channels
so far. Putative Gαi-binding sites, however, were presented: K715 and R716. Cryo-EM locates both
residues in the connecting-helix. Figure 6. The homologous position of TRPC4 to G652 of TRPC3. (A) Glycine residue in tetrameric
structure. Green domain: pore helices; Red sticks: glycine residues. (B) Left panel: enlarged view of pore
structure and glycine. Subunits behind and in front of the paper is deliberately deleted for viewer’s
preference. Right panel: top-view of pore-helices and glycine residue. Figure 6. The homologous position of TRPC4 to G652 of TRPC3. (A) Glycine residue in tetrameric
structure. Green domain: pore helices; Red sticks: glycine residues. (B) Left panel: enlarged view of pore
structure and glycine. Subunits behind and in front of the paper is deliberately deleted for viewer’s
preference. Right panel: top-view of pore-helices and glycine residue. 7. Englerin-A Mediated Gating Mechanism Englerin-A opens TRPC4/4, 5/5, 1/4 and 1/5 channels in extracellular side. Nanomolar concentration
of Englerin-A is sufficient to activate the channels. As previously mentioned, there were no agonists with
high specificity to TRPC1, 4, 5 channels. The development of this pharmacological treasure bestowed
great utility for channel research especially in discrimination of G-protein signal transduction pathway. Depending on literatures, maximum conductance was reached within 10 to 20 s after Englerin-A was
applied. This suggests that the drug action could be direct to the channel. Although it is unlikely,
whether the increase of conductance in whole-cell current level is due to increased surface level or
not is unknown. In outside-out patch, the increase in single channel conductance by Englerin-A was
reported [48]. Nonetheless, domain-specific study for mechanism of action of Englerin-A is still limited. p
p
y
g
One study addressed possible interaction sites of TRPC5 channels with Englerin-A [85]. Among the
lists of mutants targeted for acid-based sensing domain of the channel, Jeong et al. found that K554, Cells 2020, 9, 73 18 of 24 H594, and E598 of TRPC5 are putative binding sites for Englerin-A [85]. These residues are all located
in pore-helices. From the proximity per se, Englerin-A may open the channel by changing conformation
of the upper gate near selectivity filter. pp
g
y
Based on previously reported putative Englerin-A binding sites, we calculated possible binding
models. The Q386-H394 region had to be constructed since Cryo-EM structure of TRPC5 lacks such
residues. Besides K554, K594, and E598, the additional constraint to the simulation was applied that
extracellular S1-S2 linker and S3-S4 linker has nonzero likelihood of Englerin-A binding. As a primitive
result, the model predicted that both in TRPC4 and TRPC5 there is a strong cooperativity between
cis-positioned subunits. Therefore, we added additional constraint to the model that Englerin-A binds
to TRPC4 channel via K550 in A subunit and H590, E594 in B subunit. Likewise, K554 in B subunit
and H594, E598 in A subunit was selected as binding sites for TRPC5. The model suggests that the
way TRPC4 and TRPC5 channels bind with Englerin-A is different (Figure 7). It also vests great
significance to hydrophobic interaction (V, F, A, Y) in addition to acid-base sensing residues: K, H, and
E. Last but not least, the prediction index showed different amino acids for highest weighting in TRPC1. 8. Concluding Remarks Structural revelation of TRPC channels has clarified the issues or the controversies against the
function of the channels. Still, there are a few challenging problems that need to be solved. One of the
limitations is that given structures all show only closed state of the channels. We do not know yet
how exactly the selectivity filter of the channel exerts its action to conducting ions. It is obscure which
residues within the pore are responsible for stabilization of dehydrated or hydrated conducting ions. Neither the mean occupancy number of conducting ions nor whether the pore profile is multi-ionic or
not is known. Hopefully, the exact roadmap for conformational change of the gates could be addressed
in a near future. Structural analysis of the inward-rectification mechanism of TRPC4 and 5 channels
will also be intriguing. Crystal structure of Mg2+-bound and polyamine-bound inwardly-rectifying
potassium channels could be models for the study [92]. In terms of tetramerization, solving Cryo-EM structure of TRPC1/4 and TRPC1/5 channels
has substantial physiological meaning since incorporation of TRPC1 to TRPC4 and TRPC5 yields
reduced membrane excitability and calcium permeability [93]. Identifying the time and location of
heteromerization process is rather hard to answer with structural methods. Additional functional
studies targeted for protein trafficking process could show promising results. As we have discussed, cysteine residues in the S5-S6 linker hold a number of issues. Finding consistent explanation for inconsistent results would be challenging, indeed. Opening
of the channels by altering cysteines using extracellular reducing agent may constitute an independent
gating mechanism aside from gating through the G-protein pathway or Englerin-A. It is imperative to
solve a structure for channels opened by cysteine modification. The most classical activation mechanism of TRPC channels would be undoubtedly a G-protein
mediated gating. The emerging role of DAG in such gating process requires DAG binding pocket
within not only TRPC3 and 6 channels but also in TRPC4 and 5 channels. The putative binding site
predicted by modeling accompanied meaningful electrophysiological results, hence could be dearly
appreciated. The G602 site may give a useful initial-value for the equation. What makes it hard to
find structure–function relationship in G-protein gating mechanism is that the quintessential domains
such as the PDZ-domain, alternative splicing site, and PKC phosphorylation site are all located in the
far-south domain. A structural flexibility is a foe to Cryo-EM, hence truncation of flexible domains is
usual prerequisite in sample preparation step. 7. Englerin-A Mediated Gating Mechanism The sequence alignment along the pore-region shows significant difference between TRPC4, 5, and
TRPC1. Therefore, binding mechanism could be different in heteromeric channel and in homomeric
channel. Structure of TRPC1/4 or 1/5 heteromeric channel would clarify the issue. Figure 7. Binding models between Englerin-A and TRPC4, 5 channels. (A) Binding model for TRPC4. (B) Binding model for TRPC5. The model claims that not only the hydrogen bonding between hydroxyl
group and polar residues but hydrophobic interaction is important in both cases. Yellow sticks, surfaces:
subunit A; Red sticks, surfaces: subunit B; Blue sticks: Englerin-A (C) Electrophysiological recordings
of TRPC5 with various point-mutations. Basal current of TRPC5 (black bar) and peak current (white
bar) after extracellular application of 100 nM Englerin-A. (D) Sequence alignment of TRPC1, 4, and 5
channels along pore regions. TRPC1 shows significant difference while the TRPC4 and 5 channel share
great similarity. Figure 7. Binding models between Englerin-A and TRPC4, 5 channels. (A) Binding model for TRPC4. (B) Binding model for TRPC5. The model claims that not only the hydrogen bonding between hydroxyl
group and polar residues but hydrophobic interaction is important in both cases. Yellow sticks, surfaces:
subunit A; Red sticks, surfaces: subunit B; Blue sticks: Englerin-A (C) Electrophysiological recordings
of TRPC5 with various point-mutations. Basal current of TRPC5 (black bar) and peak current (white
bar) after extracellular application of 100 nM Englerin-A. (D) Sequence alignment of TRPC1, 4, and 5
channels along pore regions. TRPC1 shows significant difference while the TRPC4 and 5 channel share
great similarity. Cells 2020, 9, 73 19 of 24 19 of 24 Author Contributions: J.K. (Jinsung Kim) wrote the review; J.K. (Juyeon Ko) performed the literature searches;
J.K. (Jinsung Kim) and J.K. (Juyeon Ko) designed the structure of the article and drew the figures; C.H. drew the 8. Concluding Remarks As a functional aspect, however, flexibility of domain
may reflect functional significance. It is difficult to picture a key domain for gating in an immovable
position provided that opening and closing of the channel is a highly dynamic process. Nonetheless,
structures with minimal information of FSD would be greatly helpful to elaborate G-protein mediated
channel gating process. Finding PIP2 binding site within the channel would be equally important as to understand the
channel function and for pathophysiological implication. Disturbance in PIP2 governance to TRPC
channels could result in catastrophic collapse in intracellular calcium homeostasis since PIP2 depletion
seems to be an effective negative feedback to direct activation of Gαq-protein. In a sense, a search
for binding site to Gαq-protein within the channels is necessary. Structures for Gαi-bound TRPC4 or
TRPC5 might be found faster than Gαq since putative binding sites are already known. Although it
was X-ray crystallography that solved the structure rather than Cryo-EM, a study of Gβγ-bound GIRK
(G-protein activated Inwardly-Rectifying K channel) could be viewed as an excellent reference [94]. The development of Englerin-A has delivered unprecedented pharmacological utility. A lot of
studies analyzed functional consequences of ‘Englerin-A activated’ TRPC channels in numerous
experimental conditions. It also implies that such outcomes are not free of confounding-bias unless the
exact mechanism of the drug is addressed. We believe that fast evolving Cryo-EM will bear a fruit in
the near future and that mechanistic study for Englerin-A in parallel will definitely accelerate the pace. Author Contributions: J.K. (Jinsung Kim) wrote the review; J.K. (Juyeon Ko) performed the literature searches;
J.K. (Jinsung Kim) and J.K. (Juyeon Ko) designed the structure of the article and drew the figures; C.H. drew the Cells 2020, 9, 73 20 of 24 20 of 24 figures and revised the writings of the manuscript; I.S. edited the article and supervised the entire process of the
writing. All authors have read and agreed to the published version of the manuscript. figures and revised the writings of the manuscript; I.S. edited the article and supervised the entire process of the
writing. All authors have read and agreed to the published version of the manuscript. Funding: This research was supported by the National Research Foundation of Korea, which is funded by the
Ministry of Science, ICT (Information & Communication Technology), and Future Planning (MSIP) of the Korean
Government (2015R1A6A3A04058395 to C. Hong and 2018R1A4A1023822 to I. References 1. Nilius, B. TRP channels in disease. Biochim. Biophys. Acta 2007, 1772, 805–812. [CrossRef] [PubMed] 2. Freichel, M.;
Tsvilovskyy, V.;
Camacho-Londono, J.E. TRPC4
Handb. Exp. Pharmacol. 2014, 222, 85–128. [CrossRef] [PubMed] 2. Freichel, M.;
Tsvilovskyy, V.;
Camacho-Londono, J.E. TRPC4- and TRPC4-containing channels. Handb. Exp. Pharmacol. 2014, 222, 85–128. [CrossRef] [PubMed] 3. Zholos, A.V. Trpc5. Handb. Exp. Pharmacol. 2014, 222, 129–156. [CrossRef] [PubMed] 4. Flockerzi, V. An introduction on TRP channels. Handb. Exp. Pharmacol. 2007, 1–19. [CrossRef] 5. Montell, C.; Birnbaumer, L.; Flockerzi, V.; Bindels, R.J.; Bruford, E.A.; Caterina, M.J.; Clapham, D.E.;
Harteneck, C.; Heller, S.; Julius, D.; et al. A unified nomenclature for the superfamily of TRP cation channels. Mol. Cell 2002, 9, 229–231. [CrossRef] 6. Minard, A.; Bauer, C.C.; Wright, D.J.; Rubaiy, H.N.; Muraki, K.; Beech, D.J.; Bon, R.S. Remarkable Progress
with Small-Molecule Modulation of TRPC1/4/5 Channels: Implications for Understanding the Channels in
Health and Disease. Cells 2018, 7, 52. [CrossRef] [PubMed] 7. Greka, A.; Navarro, B.; Oancea, E.; Duggan, A.; Clapham, D.E. TRPC5 is a regulator of hippocampal neurite
length and growth cone morphology. Nat. Neurosci. 2003, 6, 837–845. [CrossRef] 8. Faber, E.S.; Sedlak, P.; Vidovic, M.; Sah, P. Synaptic activation of transient receptor potential channels by
metabotropic glutamate receptors in the lateral amygdala. Neuroscience 2006, 137, 781–794. [CrossRef] 9. Goel, M.; Sinkins, W.G.; Schilling, W.P. Selective association of TRPC channel subunits in rat brain
synaptosomes. J. Biol. Chem. 2002, 277, 48303–48310. [CrossRef] 10. Riccio, A.; Medhurst, A.D.; Mattei, C.; Kelsell, R.E.; Calver, A.R.; Randall, A.D.; Benham, C.D.; Pangalos, M.N. mRNA distribution analysis of human TRPC family in CNS and peripheral tissues. Brain Res. Mol. Brain Res. 2002, 109, 95–104. [CrossRef] 11. Okada, T.; Shimizu, S.; Wakamori, M.; Maeda, A.; Kurosaki, T.; Takada, N.; Imoto, K.; Mori, Y. Molecular cloning and functional characterization of a novel receptor-activated TRP Ca2+ channel from
mouse brain. J. Biol. Chem. 1998, 273, 10279–10287. [CrossRef] [PubMed] 12. Sossey-Alaoui, K.; Lyon, J.A.; Jones, L.; Abidi, F.E.; Hartung, A.J.; Hane, B.; Schwartz, C.E.; Stevenson, R.E.;
Srivastava, A.K. Molecular cloning and characterization of TRPC5 (HTRP5), the human homologue of a
mouse brain receptor-activated capacitative Ca2+ entry channel. Genomics 1999, 60, 330–340. [CrossRef]
[PubMed] 13. Hong, C.; Seo, H.; Kwak, M.; Jeon, J.; Jang, J.; Jeong, E.M.; Myeong, J.; Hwang, Y.J.; Ha, K.; Kang, M.J.; et al. Increased TRPC5 glutathionylation contributes to striatal neuron loss in Huntington’s disease. Brain 2015,
138, 3030–3047. [CrossRef] [PubMed] 14. 8. Concluding Remarks So) and by the Education and
Research Encouragement Fund of Seoul National University Hospital (I. So). J. Ko was supported by the BK plus
program from the MSIP. Acknowledgments: We thank Jungeun Lee for her excellent language editing. Acknowledgments: We thank Jungeun Lee for her excellent language editing. Acknowledgments: We thank Jungeun Lee for her excellent language editing. Conflicts of Interest: The authors declare no conflict of interest. Conflicts of Interest: The authors declare no conflict of interest. References Phelan, K.D.; Mock, M.M.; Kretz, O.; Shwe, U.T.; Kozhemyakin, M.; Greenfield, L.J.; Dietrich, A.;
Birnbaumer, L.; Freichel, M.; Flockerzi, V.; et al. Heteromeric canonical transient receptor potential 1
and 4 channels play a critical role in epileptiform burst firing and seizure-induced neurodegeneration. Mol. Pharmacol. 2012, 81, 384–392. [CrossRef] 15. Strubing, C.; Krapivinsky, G.; Krapivinsky, L.; Clapham, D.E. Formation of novel TRPC channels by complex
subunit interactions in embryonic brain. J. Biol. Chem. 2003, 278, 39014–39019. [CrossRef] 16. Song, X.; Zhao, Y.; Narcisse, L.; Duffy, H.; Kress, Y.; Lee, S.; Brosnan, C.F. Canonical transient receptor
potential channel 4 (TRPC4) co-localizes with the scaffolding protein ZO-1 in human fetal astrocytes in
culture. Glia 2005, 49, 418–429. [CrossRef] 21 of 24 21 of 24 Cells 2020, 9, 73 17. Phelan, K.D.; Shwe, U.T.; Abramowitz, J.; Wu, H.; Rhee, S.W.; Howell, M.D.; Gottschall, P.E.; Freichel, M.;
Flockerzi, V.; Birnbaumer, L.; et al. Canonical transient receptor channel 5 (TRPC5) and TRPC1/4 contribute
to seizure and excitotoxicity by distinct cellular mechanisms. Mol. Pharmacol. 2013, 83, 429–438. [CrossRef] 18. Hui, H.; McHugh, D.; Hannan, M.; Zeng, F.; Xu, S.Z.; Khan, S.U.; Levenson, R.; Beech, D.J.; Weiss, J.L. Calcium-sensing mechanism in TRPC5 channels contributing to retardation of neurite outgrowth. J. Physiol. 2006, 572, 165–172. [CrossRef] 19. Lee, K.P.; Jun, J.Y.; Chang, I.Y.; Suh, S.H.; So, I.; Kim, K.W. TRPC4 is an essential component of the nonselective
cation channel activated by muscarinic stimulation in mouse visceral smooth muscle cells. Mol. Cells 2005,
20, 435–441. 20. Sakamoto, T.; Unno, T.; Kitazawa, T.; Taneike, T.; Yamada, M.; Wess, J.; Nishimura, M.; Komori, S. Three distinct
muscarinic signalling pathways for cationic channel activation in mouse gut smooth muscle cells. J. Physiol. 2007, 582, 41–61. [CrossRef] 21. Dresviannikov, A.V.; Bolton, T.B.; Zholos, A.V. Muscarinic receptor-activated cationic channels in murine
ileal myocytes. Br. J. Pharmacol. 2006, 149, 179–187. [CrossRef] [PubMed] 22. Tsvilovskyy, V.V.; Zholos, A.V.; Aberle, T.; Philipp, S.E.; Dietrich, A.; Zhu, M.X.; Birnbaumer, L.; Freichel, M.;
Flockerzi, V. Deletion of TRPC4 and TRPC6 in mice impairs smooth muscle contraction and intestinal motility
in vivo. Gastroenterology 2009, 137, 1415–1424. [CrossRef] [PubMed] 23. De Clercq, K.; Held, K.; Van Bree, R.; Meuleman, C.; Peeraer, K.; Tomassetti, C.; Voets, T.; D’Hooghe, T.;
Vriens, J. Functional expression of transient receptor potential channels in human endometrial stromal cells
during the luteal phase of the menstrual cycle. Hum. Reprod. 2015, 30, 1421–1436. [CrossRef] [PubMed] 24. References Beck, A.; Gotz, V.; Qiao, S.; Weissgerber, P.; Flockerzi, V.; Freichel, M.; Boehm, U. Functional Characterization
of Transient Receptor Potential (TRP) Channel C5 in Female Murine Gonadotropes. Endocrinology 2017, 158,
887–902. [CrossRef] 25. Persoons, E.; Hennes, A.; De Clercq, K.; Van Bree, R.; Vriens, G.; O, D.F.; Peterse, D.; Vanhie, A.; Meuleman, C.;
Voets, T.; et al. Functional Expression of TRP Ion Channels in Endometrial Stromal Cells of Endometriosis
Patients. Int. J. Mol. Sci. 2018, 19, 2467. [CrossRef] 26. De Clercq, K.; Van den Eynde, C.; Hennes, A.; Van Bree, R.; Voets, T.; Vriens, J. The functional expression of
transient receptor potential channels in the mouse endometrium. Hum. Reprod. 2017, 32, 615–630. [CrossRef] 27. Jung, C.; Gene, G.G.; Tomas, M.; Plata, C.; Selent, J.; Pastor, M.; Fandos, C.; Senti, M.; Lucas, G.; Elosua, R.; et al. A gain-of-function SNP in TRPC4 cation channel protects against myocardial infarction. Cardiovasc. Res. 2011, 91, 465–471. [CrossRef] 28. Liu, F.F.; Ma, Z.Y.; Li, D.L.; Feng, J.B.; Zhang, K.; Wang, R.; Zhang, W.; Li, L.; Zhang, Y. Differential expression
of TRPC channels in the left ventricle of spontaneously hypertensive rats. Mol. Biol. Rep. 2010, 37, 2645–2651. [CrossRef] 29. Kojima, A.; Kitagawa, H.; Omatsu-Kanbe, M.; Matsuura, H.; Nosaka, S. Ca2+ paradox injury mediated
through TRPC channels in mouse ventricular myocytes. Br. J. Pharmacol. 2010, 161, 1734–1750. [CrossRef] 30. Poteser, M.; Graziani, A.; Rosker, C.; Eder, P.; Derler, I.; Kahr, H.; Zhu, M.X.; Romanin, C.; Groschner, K. TRPC3 and TRPC4 associate to form a redox-sensitive cation channel. Evidence for expression of native
TRPC3-TRPC4 heteromeric channels in endothelial cells. J. Biol. Chem. 2006, 281, 13588–13595. [CrossRef] 31. Freichel, M.; Suh, S.H.; Pfeifer, A.; Schweig, U.; Trost, C.; Weissgerber, P.; Biel, M.; Philipp, S.; Freise, D.;
Droogmans, G.; et al. Lack of an endothelial store-operated Ca2+ current impairs agonist-dependent
vasorelaxation in TRP4-/- mice. Nat. Cell Biol. 2001, 3, 121–127. [CrossRef] [PubMed] 32. Fantozzi, I.; Zhang, S.; Platoshyn, O.; Remillard, C.V.; Cowling, R.T.; Yuan, J.X. Hypoxia increases AP-1
binding activity by enhancing capacitative Ca2+ entry in human pulmonary artery endothelial cells. Am. J. Physiol. Lung Cell Mol. Physiol. 2003, 285, L1233–L1245. [CrossRef] [PubMed] 33. Murata, T.; Lin, M.I.; Stan, R.V.; Bauer, P.M.; Yu, J.; Sessa, W.C. Genetic evidence supporting caveolae
microdomain regulation of calcium entry in endothelial cells. J. Biol. Chem. 2007, 282, 16631–16643. [CrossRef] [PubMed] 34. Kim, J.; Ko, J.; Myeong, J.; Kwak, M.; Hong, C.; So, I. References TRPC1 as a negative regulator for TRPC4 and TRPC5
channels. Pflug. Arch. 2019, 471, 1045–1053. [CrossRef] [PubMed] 35. Zholos, A.V.; Zholos, A.A.; Bolton, T.B. G-protein-gated TRP-like cationic channel activated by muscarinic
receptors: Effect of potential on single-channel gating. J. Gen. Physiol. 2004, 123, 581–598. [CrossRef]
[PubMed] 22 of 24 Cells 2020, 9, 73 36. Mery, L.; Magnino, F.; Schmidt, K.; Krause, K.H.; Dufour, J.F. Alternative splice variants of hTrp4 differentially
interact with the C-terminal portion of the inositol 1,4,5-trisphosphate receptors. FEBS Lett. 2001, 487,
377–383. [CrossRef] 7. Ko, J.; Myeong, J.; Shin, Y.C.; So, I. Differential PI(4,5)P2 sensitivities of TRPC4, C5 homomeric and TRPC
C1/5 heteromeric channels. Sci. Rep. 2019, 9, 1849. [CrossRef] 38. Myeong, J.; Ko, J.; Kwak, M.; Kim, J.; Woo, J.; Ha, K.; Hong, C.; Yang, D.; Kim, H.J.; Jeon, J.H.; et al. Dual action
of the Galphaq-PLCbeta-PI(4,5)P2 pathway on TRPC1/4 and TRPC1/5 heterotetramers. Sci. Rep. 2018, 8,
12117. [CrossRef] 39. Kim, H.; Jeon, J.P.; Hong, C.; Kim, J.; Myeong, J.; Jeon, J.H.; So, I. An essential role of PI(4,5)P(2) for
maintaining the activity of the transient receptor potential canonical (TRPC)4beta. Pflug. Arch. 2013, 465,
1011–1021. [CrossRef] 40. Schaefer, M.; Plant, T.D.; Stresow, N.; Albrecht, N.; Schultz, G. Functional differences between TRPC4 splice
variants. J. Biol. Chem. 2002, 277, 3752–3759. [CrossRef] 41. Kim, B.J.; Kim, M.T.; Jeon, J.H.; Kim, S.J.; So, I. Involvement of phosphatidylinositol 4,5-bisphosphate in the
desensitization of canonical transient receptor potential 5. Biol. Pharm. Bull. 2008, 31, 1733–1738. [CrossRef]
[PubMed] 42. Otsuguro, K.; Tang, J.; Tang, Y.; Xiao, R.; Freichel, M.; Tsvilovskyy, V.; Ito, S.; Flockerzi, V.; Zhu, M.X.;
Zholos, A.V. Isoform-specific inhibition of TRPC4 channel by phosphatidylinositol 4,5-bisphosphate. J. Biol. Chem. 2008, 283, 10026–10036. [CrossRef] [PubMed] 43. Zhu, M.H.; Chae, M.; Kim, H.J.; Lee, Y.M.; Kim, M.J.; Jin, N.G.; Yang, D.K.; So, I.; Kim, K.W. Desensitization of
canonical transient receptor potential channel 5 by protein kinase C. Am. J. Physiol. Cell Physiol. 2005, 289,
C591–G600. [CrossRef] [PubMed] 44. Hofmann, T.; Obukhov, A.G.; Schaefer, M.; Harteneck, C.; Gudermann, T.; Schultz, G. Direct activation of
human TRPC6 and TRPC3 channels by diacylglycerol. Nature 1999, 397, 259–263. [CrossRef] 45. Jeon, J.P.; Hong, C.; Park, E.J.; Jeon, J.H.; Cho, N.H.; Kim, I.G.; Choe, H.; Muallem, S.; Kim, H.J.; So, I. Selective Galphai subunits as novel direct activators of transient receptor potential canonical (TRPC)4 and
TRPC5 channels. J. Biol. Chem. 2012, 287, 17029–17039. [CrossRef] 46. 56.
Tang, Q.; Guo, W.; Zheng, L.; Wu, J.X.; Liu, M.; Zhou, X.; Zhang, X.; Chen, L. Structure of the receptor-activated
human TRPC6 and TRPC3 ion channels. Cell Res. 2018, 28, 746–755. [CrossRef] References [CrossRef] 23 of 24 23 of 24 Cells 2020, 9, 73 57. Lin, Z.; Chen, Q.; Lee, M.; Cao, X.; Zhang, J.; Ma, D.; Chen, L.; Hu, X.; Wang, H.; Wang, X.; et al. Exome sequencing reveals mutations in TRPV3 as a cause of Olmsted syndrome. Am. J. Hum. Genet. 2012,
90, 558–564. [CrossRef] 58. Teng, J.; Loukin, S.H.; Anishkin, A.; Kung, C. L596-W733 bond between the start of the S4-S5 linker and
the TRP box stabilizes the closed state of TRPV4 channel. Proc. Natl. Acad. Sci. USA 2015, 112, 3386–3391. [CrossRef] 59. Kim, J.; Kwak, M.; Jeon, J.P.; Myeong, J.; Wie, J.; Hong, C.; Kim, S.Y.; Jeon, J.H.; Kim, H.J.; So, I. Isoform- and
receptor-specific channel property of canonical transient receptor potential (TRPC)1/4 channels. Pflug. Arch. 2014, 466, 491–504. [CrossRef] 60. Dosey, T.L.; Wang, Z.; Fan, G.; Zhang, Z.; Serysheva, I.I.; Chiu, W.; Wensel, T.G. Structures of TRPV2 in
distinct conformations provide insight into role of the pore turret. Nat. Struct. Mol. Biol. 2019, 26, 40–49. [CrossRef] 61. Zubcevic, L.; Herzik, M.A., Jr.; Chung, B.C.; Liu, Z.; Lander, G.C.; Lee, S.Y. Cryo-electron microscopy structure
of the TRPV2 ion channel. Nat. Struct. Mol. Biol. 2016, 23, 180–186. [CrossRef] [PubMed] 62. Dietrich, A.; Fahlbusch, M.; Gudermann, T. Classical Transient Receptor Potential 1 (TRPC1
Channel Regulator? Cells 2014, 3, 939–962. [CrossRef] [PubMed] 3. Obukhov, A.G.; Nowycky, M.C. A cytosolic residue mediates Mg2+ block and regulates inward cur
amplitude of a transient receptor potential channel. J. Neurosci. 2005, 25, 1234–1239. [CrossRef] [PubM 64. Ishihara, K.; Mitsuiye, T.; Noma, A.; Takano, M. The Mg2+ block and intrinsic gating underlying inward
rectification of the K+ current in guinea-pig cardiac myocytes. J. Physiol. 1989, 419, 297–320. [CrossRef] 65. Matsuda, H.; Saigusa, A.; Irisawa, H. Ohmic conductance through the inwardly rectifying K channel and
blocking by internal Mg2+. Nature 1987, 325, 156–159. [CrossRef] 66. Stanfield, P.R.; Davies, N.W.; Shelton, P.A.; Sutcliffe, M.J.; Khan, I.A.; Brammar, W.J.; Conley, E.C. A single
aspartate residue is involved in both intrinsic gating and blockage by Mg2+ of the inward rectifier, IRK1. J. Physiol. 1994, 478, 1–6. [CrossRef] 67. Fakler, B.; Brandle, U.; Glowatzki, E.; Weidemann, S.; Zenner, H.P.; Ruppersberg, J.P. Strong voltage-dependent
inward rectification of inward rectifier K+ channels is caused by intracellular spermine. Cell 1995, 80, 149–154. [CrossRef] 68. Lopatin, A.N.; Makhina, E.N.; Nichols, C.G. Potassium channel block by cytoplasmic polyamines as the
mechanism of intrinsic rectification. References Jeon, J.P.; Lee, K.P.; Park, E.J.; Sung, T.S.; Kim, B.J.; Jeon, J.H.; So, I. The specific activation of TRPC
protein subtype. Biochem. Biophys. Res. Commun. 2008, 377, 538–543. [CrossRef] 47. Miller, M.; Shi, J.; Zhu, Y.; Kustov, M.; Tian, J.B.; Stevens, A.; Wu, M.; Xu, J.; Long, S.; Yang, P.; et al. Identification of ML204, a novel potent antagonist that selectively modulates native TRPC4/C5 ion channels. J. Biol. Chem. 2011, 286, 33436–33446. [CrossRef] 48. Akbulut, Y.; Gaunt, H.J.; Muraki, K.; Ludlow, M.J.; Amer, M.S.; Bruns, A.; Vasudev, N.S.; Radtke, L.; Willot, M.;
Hahn, S.; et al. (-)-Englerin A is a potent and selective activator of TRPC4 and TRPC5 calcium channels. Angew Chem. Int. Ed. Engl. 2015, 54, 3787–3791. [CrossRef] 49. Rubaiy, H.N.; Ludlow, M.J.; Bon, R.S.; Beech, D.J. Pico145—Powerful new tool for TRPC1/4/5 channels. Channels (Austin) 2017, 11, 362–364. [CrossRef] 50. Kuhlbrandt, W. Biochemistry. The resolution revolution. Science 2014, 343, 1443–1444. [CrossRef] 51. Duan, J.; Li, J.; Chen, G.L.; Ge, Y.; Liu, J.; Xie, K.; Peng, X.; Zhou, W.; Zhong, J.; Zhang, Y.; et al. Cryo-EM structure of TRPC5 at 2.8-A resolution reveals unique and conserved structural elements essential
for channel function. Sci. Adv. 2019, 5, eaaw7935. [CrossRef] [PubMed] 52. Vinayagam, D.; Mager, T.; Apelbaum, A.; Bothe, A.; Merino, F.; Hofnagel, O.; Gatsogiannis, C.; Raunser, S. Electron cryo-microscopy structure of the canonical TRPC4 ion channel. Elife 2018, 7. [CrossRef] [PubMed] 52. Vinayagam, D.; Mager, T.; Apelbaum, A.; Bothe, A.; Merino, F.; Hofnagel, O.; Gatsogiannis, C.; Raunser, S. Electron cryo-microscopy structure of the canonical TRPC4 ion channel. Elife 2018, 7. [CrossRef] [PubMed]
53. Duan, J.; Li, J.; Zeng, B.; Chen, G.L.; Peng, X.; Zhang, Y.; Wang, J.; Clapham, D.E.; Li, Z.; Zhang, J. Structure of 53. Duan, J.; Li, J.; Zeng, B.; Chen, G.L.; Peng, X.; Zhang, Y.; Wang, J.; Clapham, D.E.; Li, Z.; Zhang, J. Structure of
the mouse TRPC4 ion channel. Nat. Commun. 2018, 9, 3102. [CrossRef] [PubMed] 54. Long, S.B.; Campbell, E.B.; Mackinnon, R. Voltage sensor of Kv1.2: Structural basis of electromechanical
coupling. Science 2005, 309, 903–908. [CrossRef] 55. Fan, C.; Choi, W.; Sun, W.; Du, J.; Lu, W. Structure of the human lipid-gated cation channel TRPC3. Elife 2018, 7. [CrossRef] 56. Tang, Q.; Guo, W.; Zheng, L.; Wu, J.X.; Liu, M.; Zhou, X.; Zhang, X.; Chen, L. Structure of the receptor-activated
human TRPC6 and TRPC3 ion channels. Cell Res. 2018, 28, 746–755. References Nature 1994, 372, 366–369. [CrossRef] 69. Lopatin, A.N.; Makhina, E.N.; Nichols, C.G. The mechanism of inward rectification of potassium channels:
“long-pore plugging” by cytoplasmic polyamines. J. Gen. Physiol. 1995, 106, 923–955. [CrossRef] 70. Pegg, A.E. The function of spermine. IUBMB Life 2014, 66, 8–18. [CrossRef] 71. Tsvilovskyy, V.V.; Zholos, A.V.; Bolton, T.B. Effects of polyamines on the muscarinic receptor-operated
cation current in guinea-pig ileal smooth muscle myocytes. Br. J. Pharmacol. 2004, 143, 968–975. [CrossRef]
[PubMed] 72. Kim, J.; Moon, S.H.; Shin, Y.C.; Jeon, J.H.; Park, K.J.; Lee, K.P.; So, I. Intracellular spermine blocks TRPC4
channel via electrostatic interaction with C-terminal negative amino acids. Pflug. Arch. 2016, 468, 551–561. [CrossRef] [PubMed] 73. Kim, J.; Moon, S.H.; Kim, T.; Ko, J.; Jeon, Y.K.; Shin, Y.C.; Jeon, J.H.; So, I. Analysis of interaction between
intracellular spermine and transient receptor potential canonical 4 channel; multiple candidate sites of
negatively charged amino acids for the inward rectification of transient receptor potential canonical 4. Korean J. Phiysiol. Pharmacol. 2019, in press. 74. Hofmann, T.; Schaefer, M.; Schultz, G.; Gudermann, T. Subunit composition of mammalian transient receptor
potential channels in living cells. Proc. Natl. Acad. Sci. USA 2002, 99, 7461–7466. [CrossRef] [PubMed] 74. Hofmann, T.; Schaefer, M.; Schultz, G.; Gudermann, T. Subunit composition of mammalian transient receptor
potential channels in living cells. Proc. Natl. Acad. Sci. USA 2002, 99, 7461–7466. [CrossRef] [PubMed]
75. Jeon, J.P.; Roh, S.E.; Wie, J.; Kim, J.; Kim, H.; Lee, K.P.; Yang, D.; Jeon, J.H.; Cho, N.H.; Kim, I.G.; et al. 75. Jeon, J.P.; Roh, S.E.; Wie, J.; Kim, J.; Kim, H.; Lee, K.P.; Yang, D.; Jeon, J.H.; Cho, N.H.; Kim, I.G.; et al. Activation of TRPC4beta by Galphai subunit increases Ca2+ selectivity and controls neurite morphogenesis
in cultured hippocampal neuron. Cell Calcium 2013, 54, 307–319. [CrossRef] [PubMed] 76. Schindl, R.; Frischauf, I.; Kahr, H.; Fritsch, R.; Krenn, M.; Derndl, A.; Vales, E.; Muik, M.; Derler, I.;
Groschner, K.; et al. The first ankyrin-like repeat is the minimum indispensable key structure for functional
assembly of homo- and heteromeric TRPC4/TRPC5 channels. Cell Calcium 2008, 43, 260–269. [CrossRef]
[PubMed] 24 of 24 24 of 24 Cells 2020, 9, 73 77. Lepage, P.K.; Lussier, M.P.; McDuff, F.O.; Lavigne, P.; Boulay, G. The self-association of two N-terminal
interaction domains plays an important role in the tetramerization of TRPC4. Cell Calcium 2009, 45, 251–259. [CrossRef] 78. Myeong, J.; Kwak, M.; Hong, C.; Jeon, J.H.; So, I. References Identification of a membrane-targeting domain of the
transient receptor potential canonical (TRPC)4 channel unrelated to its formation of a tetrameric structure. J. Biol. Chem. 2014, 289, 34990–35002. [CrossRef] 79. Hong, C.; Kwak, M.; Myeong, J.; Ha, K.; Wie, J.; Jeon, J.H.; So, I. Extracellular disulfide bridges stabilize
TRPC5 dimerization, trafficking, and activity. Pflug. Arch. 2015, 467, 703–712. [CrossRef] 80. Myeong, J.; Ko, J.; Hong, C.; Yang, D.; Lee, K.P.; Jeon, J.H.; So, I. The interaction domains of transient receptor
potential canonical (TRPC)1/4 and TRPC1/5 heteromultimeric channels. Biochem. Biophys. Res. Commun. 2016, 474, 476–481. [CrossRef] 81. Sevier, C.S.; Kaiser, C.A. Formation and transfer of disulphide bonds in living cells. Nat. Rev. Mol. Cell Biol. 2002, 3, 836–847. [CrossRef] 82. Hille, B. Ionic Channels of Excitable Membranes; Sinauer Associates: Sunderland, MA, USA, 2001. 83. Yoshida, T.; Inoue, R.; Morii, T.; Takahashi, N.; Yamamoto, S.; Hara, Y.; Tominaga, M.; Shimizu, S.; Sato, Y.;
Mori, Y. Nitric oxide activates TRP channels by cysteine S-nitrosylation. Nat. Chem. Biol. 2006, 2, 596–607. [CrossRef] [PubMed] 84. Xu, S.Z.; Sukumar, P.; Zeng, F.; Li, J.; Jairaman, A.; English, A.; Naylor, J.; Ciurtin, C.; Majeed, Y.; Milligan, C.J.;
et al. TRPC channel activation by extracellular thioredoxin. Nature 2008, 451, 69–72. [CrossRef] [PubMed] 84. Xu, S.Z.; Sukumar, P.; Zeng, F.; Li, J.; Jairaman, A.; English, A.; Naylor, J.; Ciurtin, C.; Majeed, Y.; Milligan, C.J.;
et al. TRPC channel activation by extracellular thioredoxin. Nature 2008, 451, 69–72. [CrossRef] [PubMed]
85. Jeong, S.; Ko, J.; Kim, M.; Park, K.C.; Park, E.Y.J.; Kim, J.; Baik, Y.; Wie, J.; Cho, A.E.; Jeon, J.H.; et al. et al. TRPC channel activation by extracellular thioredoxin. Nature 2008, 451, 69 72. [CrossRef] [PubMed]
85. Jeong, S.; Ko, J.; Kim, M.; Park, K.C.; Park, E.Y.J.; Kim, J.; Baik, Y.; Wie, J.; Cho, A.E.; Jeon, J.H.; et al. Englerin A-sensing charged residues for transient receptor potential canonical 5 channel activation. Korean J. Physiol. Pharmacol. 2019, 23, 191–201. [CrossRef] [PubMed] 86. Storch, U.; Forst, A.L.; Pardatscher, F.; Erdogmus, S.; Philipp, M.; Gregoritza, M.; Mederos, Y.S.M.;
Gudermann, T. Dynamic NHERF interaction with TRPC4/5 proteins is required for channel gating by
diacylglycerol. Proc. Natl. Acad. Sci. USA 2017, 114, E37–E46. [CrossRef] [PubMed] 87. Mederos, Y.S.M.; Gudermann, T.; Storch, U. Emerging Roles of Diacylglycerol-Sensitive TRPC4/5 Channels. Cells 2018, 7, 218. [CrossRef] 88. Ko, J.; Myeong, J.; Kwak, M.; Jeon, J.H.; So, I. References Identification of phospholipase C beta downstream effect
on transient receptor potential canonical 1/4, transient receptor potential canonical 1/5 channels. Korean J. Physiol. Pharmacol. 2019, 23, 357–366. [CrossRef] 89. Yang, F.; Xiao, X.; Cheng, W.; Yang, W.; Yu, P.; Song, Z.; Yarov-Yarovoy, V.; Zheng, J. Structural mechanism
underlying capsaicin binding and activation of the TRPV1 ion channel. Nat. Chem. Biol. 2015, 11, 518–524. [CrossRef] 90. Fine, M.; Schmiege, P.; Li, X. Structural basis for PtdInsP2-mediated human TRPML1 regulation. Nat. Commun. 2018, 9, 4192. [CrossRef] 91. Lichtenegger, M.; Tiapko, O.; Svobodova, B.; Stockner, T.; Glasnov, T.N.; Schreibmayer, W.; Platzer, D.;
de la Cruz, G.G.; Krenn, S.; Schober, R.; et al. An optically controlled probe identifies lipid-gating fenestrations
within the TRPC3 channel. Nat. Chem. Biol. 2018, 14, 396–404. [CrossRef] 92. Nishida, M.; MacKinnon, R. Structural basis of inward rectification: cytoplasmic pore of the G protein-gated
inward rectifier GIRK1 at 1.8 A resolution. Cell 2002, 111, 957–965. [CrossRef] 93. Storch, U.; Forst, A.L.; Philipp, M.; Gudermann, T.; Mederos y Schnitzler, M. Transient receptor potential
channel 1 (TRPC1) reduces calcium permeability in heteromeric channel complexes. J. Biol. Chem. 2012, 287,
3530–3540. [CrossRef] [PubMed] 94. Whorton, M.R.; MacKinnon, R. X-ray structure of the mammalian GIRK2-betagamma G-protein complex. Nature 2013, 498, 190–197. [CrossRef] [PubMed] © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/). © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).
|
https://openalex.org/W3158795590
|
https://www.frontiersin.org/articles/10.3389/fmicb.2021.648890/pdf
|
English
| null |
Taxonomy and Phylogeny of Rust Fungi on Hamamelidaceae
|
Frontiers in microbiology
| 2,021
|
cc-by
| 6,757
|
Edited by: Edited by:
Jun-Jun Liu,
Canadian Forest Service, Natural
Resources Canada, Canada Reviewed by:
Ludwig Beenken,
Swiss Federal Institute for Forest,
Snow and Landscape Research
(WSL), Switzerland
Peng Zhao,
Chinese Academy of Sciences, China Chinese Academy of Sciences, China *Correspondence:
Yingmei Liang
liangym@bjfu.edu.cn Specialty section:
This article was submitted to
Microbe and Virus Interactions with
Plants,
a section of the journal
Frontiers in Microbiology
Received: 02 January 2021
Accepted: 11 March 2021
Published: 28 April 2021
Citation:
Liu Y, Cao B, Tian C, Ono Y, Lin W
and Liang Y (2021) Taxonomy
and Phylogeny of Rust Fungi on
Hamamelidaceae. Specialty section:
This article was submitted to
Microbe and Virus Interactions with
Plants, Specialty section:
This article was submitted to
Microbe and Virus Interactions with
Plants,
a section of the journal
Frontiers in Microbiology
Received: 02 January 2021
Accepted: 11 March 2021
Published: 28 April 2021 Keywords: Pucciniales, Puccinia, life cycle, new species and genus, evolution, Asia Taxonomy and Phylogeny of Rust
Fungi on Hamamelidaceae 1 The Key Laboratory for Silviculture and Conservation of Ministry of Education, Beijing Forestry University, Beijing, China,
2 State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China, 3 College
of Education, Ibaraki University, Ibaraki, Japan, 4 Museum of Beijing Forestry University, Beijing Forestry University, Beijing,
China Hamamelidaceae is composed of woody plant taxa of important economic value;
however, reports on diseases affecting these plants are rare. Three kinds of rusts were
studied, of which the first one is characterized by catenulate spores in peridiate columnar
sori on Sycopsis sinensis, the second one produces two-celled pedicellate teliospores in
pulvinate sori on S. sinensis and Corylopsis spp., and the last one produces aeciospores
in cup-shaped aecia on Hamamelis spp. Phylogenetic analyses indicated that the three
species belong to the same genus in Pucciniaceae. The first fungus is distinct in
teliospore morphology, producing one-celled catenulate spores in peridiate columnar
sori and molecular phylogeny from species of other genera. Thus, it is described herein
as a new genus and species Novopuccinia sycopsis-sinensis in Pucciniaceae. The
latter two species were reported as Puccinia corylopsidis and Aecidium hamamelidis,
respectively. However, phylogenetic analysis using ITS and 28S genes has revealed that
these are closely related to the new genus and species. By combining host, distribution,
and evolutionary hypothesis of rust fungi with endocyclic life cycle, these are reclassified
as N. corylopsidis and N. hamamelidis. Taxonomic descriptions, illustrations, and a key
to rust fungal species occurring in Hamamelidaceae in Asia are provided. ORIGINAL RESEARCH
published: 28 April 2021
doi: 10.3389/fmicb.2021.648890 Taxonomy and Phylogeny of Rust
Fungi on Hamamelidaceae
Yun Liu1, Bin Cao2, Chengming Tian1, Yoshitaka Ono3, Weiwei Lin1 and Yingmei Liang4*
1 The Key Laboratory for Silviculture and Conservation of Ministry of Education, Beijing Forestry University, Beijing, China,
2 State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China, 3 College
of Education, Ibaraki University, Ibaraki, Japan, 4 Museum of Beijing Forestry University, Beijing Forestry University, Beijing,
China INTRODUCTION Several Hamamelidaceae plant species are of significant ecological value in the subtropical humid
evergreen broad-leaved forest community (Xu, 2001). Some species such as Sycopsis sinensis also
have special habitat requirements and therefore show scattered distribution in the whole range
of the subtropical humid evergreen broad-leaved forests. S. sinensis has been listed as a class of
protected species in several provinces (Zang and Bian, 2003; Xu et al., 2007). p
p
p
g
Four rust fungi are known to infect trees of Hamamelidaceae in Asia. Aecidium hamamelidis
Dietel (synonym: Puccinia mitriformis S. Ito on Sasa) parasitizes Hamamelis (Hiratsuka and
Sato, 1970; Hiratsuka et al., 1992; Ono, 2017). P. sasicola (Hara) Hino & Katum is heteroecious
but demicyclic, host-alternating between Corylopsis (spermogonial/aecial stages) and Sasamorpha
(uredinial/telial stages) (Hara, 1939, 1952). P. corylopsidis Cummins with two different types of
teliospores is microcyclic on Corylopsis, and P. sakamotoi Hirats. f. & Yoshino with columnar telia
is demicyclic and autoecious on Distylium (Hiratsuka, 1942). Edited by:
Jun-Jun Liu,
Canadian Forest Service, Natural
Resources Canada, Canada
Reviewed by:
Ludwig Beenken,
Swiss Federal Institute for Forest,
Snow and Landscape Research
(WSL), Switzerland
Peng Zhao,
Chinese Academy of Sciences, China
*Correspondence:
Yingmei Liang Phylogenetic Analyses y
g
y
The sequences of the 28S and 5.8S-ITS2 genes from this study and
reference sequences obtained from GenBank (Supplementary
Table 1) were aligned and edited manually using online MAFFT3
(Katoh and Standley, 2013). The alignments were concatenated
for phylogenetic analyses. The alignments were concatenated
for phylogenetic analyses. Maximum parsimony (MP) analyses
were conducted with PAUP v. 4.0b10 (Swofford, 2003), using
1,000 heuristic search replicates with random additions of
sequences with the tree bisection and reconnection (TBR)
branch swapping algorithm. All molecular characters were
unordered and given equal weight; analyses were performed
with gaps treated as missing data. All equally parsimonious
trees found were saved in the MP analyses. Other calculated
parsimony scores were tree length (TL), consistency index
(CI), retention index (RI), and rescaled consistency (RC). MP bootstrap analyses with 1,000 replicates were performed
in the same way, with 10 rounds of replicates of heuristic
search with random addition of sequences and subsequent
TBR branch swapping during each bootstrap replicate. ML
analysis was performed with a GTR + I + G substitution
model selected by MrModeltest v.2.3 (Posada and Crandall,
1998). Branch support was evaluated with a bootstrapping
(BS) method with 1,000 replicates. For Bayesian Inference (BI),
the best-fit model of substitution among those implementable
in MrBayes was estimated separately for each gene using
jModeltest (Darriba et al., 2012) on the CIPRES portal, based
on the Bayesian Information Criterion (BIC). According to the
results of MrModeltest, Bayesian Inference (BI) of the 28S was
performed using MrBayes 3.1.2 (Ronquist and Huelsenbeck,
2003), implementing the GTR + I + G model. BI was
performed by a Markov chain Monte Carlo (MCMC) algorithm
(Rannala and Yang, 1996). The MCMC chains were run for 1
million generations and saved every 1,000 generations. Sequence
alignments were deposited at TreeBASE4 under the accession
number 27,477. 1http://www.indexfungorum.org
2http://www.efloras.org/ Study Site and Specimens Study Site and Specimens
Four rust specimens were collected from diseased S. sinensis on
Mt. Sanqingshan, northeast Jiangxi Province, China (Figure 1),
in 2017. Rust specimens on Hamamelis sp. were collected from
Ibaraki, Japan, by Y. Ono (IBAR). The host tree was identified by
referring to the Flora of China2. These specimens were deposited
at the Museum of the Beijing Forestry University (BJFC), Beijing,
China. Individual specimens are listed under each species in
the Taxonomy section. Photomicrographs of type specimens
of P. corylopsidis were provided by the U. S. National Fungus
Collections (BPI). DNA Extraction and PCR Amplification DNA Extraction and PCR Amplification
Genomic DNA was extracted from the sori by a method
described by Tian et al. (2004). The primer pairs NL1 and
NL4 for nuc rDNA 28S (28S) (O’Donnell, 1993) and Rust2inv
and ITS4rust for Nuc rDNA internal transcribed spacer region
(5.8S-ITS2) (Aime, 2006; Beenken et al., 2012) were used in
this study. The PCR conditions were fully described by Liu
et al. (2018). The PCR amplification products were estimated
visually by electrophoresis in 1.5% agarose gels. DNA sequencing
was performed using an ABI PRISM
R⃝3730xl DNA Analyzer
with BigDye
R⃝
Terminater Kit v. 3.1 (Invitrogen) at the
Shanghai Invitrogen Biological Technology Company Limited
(Beijing, China). In this study, three distinct symptoms with different
sporulation of rust fungi were observed on Hamamelidaceae
trees. Of these, two fungi produce telia; i.e., one produces two-
celled pedicellate spores in pulvinate sori on S. sinensis and
Corylopsis sp., while another generates one-celled catenulate
spores in peridiate columnar sori on S. sinensis. Long intercalary
cells are prominent in this fungus. The other produces
spermogonia and cup-shaped aecia on Hamamelis spp. This
study aimed to determine the taxonomic status and relationship
of three Hamamelidaceae fungi by morphological, molecular
phylogenetic, host, and evolutionary analyses of these endocyclic
rust fungi. A new genus, Novopuccinia, and three species, namely,
N. sycopsis-sinensis sp. nov., N. corylopsidis comb. nov., and
N. hamamelidis comb. nov., are described and presented here in
detail. All of the rust species of Hamamelidaceae are listed, and
their hosts and distribution are provided. 3http://mafft.cbrc.jp/alignment/server/
4www.treebase.org Citation: Liu Y, Cao B, Tian C, Ono Y, Lin W
and Liang Y (2021) Taxonomy
and Phylogeny of Rust Fungi on
Hamamelidaceae. Front. Microbiol. 12:648890. doi: 10.3389/fmicb.2021.648890 April 2021 | Volume 12 | Article 648890 1 Frontiers in Microbiology | www.frontiersin.org Rust Fungi on Hamamelidaceae Liu et al. Liu et al. S-3400 scanning electron microscope (Hitachi, Tokyo, Japan)
operated at 5 kV. S-3400 scanning electron microscope (Hitachi, Tokyo, Japan)
operated at 5 kV. Rust fungi, Pucciniales, are obligate plant parasites and
comprise
one
of
the
largest
groups
in
Basidiomycota. Approximately one half of the nearly 8,000 described rust
species belong to the genus Puccinia Pers., which is mainly
characterized by two-celled teliospores (Index Fungorum)1. Several molecular phylogenetic studies have shown that Puccinia
spp. is a polyphyletic group and some species are misclassified
under this genus name (Aime, 2006; Minnis et al., 2012; Beenken
and Wood, 2015). These recent studies have revealed that the
taxonomy of the species within the huge genus Puccinia is
worth exploring. RESULTS Type species: Novopuccinia sycopsis-sinensis Y. M. Liang &
Y. Liu, sp. nov. Type species: Novopuccinia sycopsis-sinensis Y. M. Liang &
Y. Liu, sp. nov. To confirm the phylogenetic placement of the fungi in
Hamamelidaceae and their relationship, a phylogenetic tree
was established based on 28S (Figure 2). The phylogenetic
results were mostly congruent with recent studies of rust fungi
(Aime and Mctaggart, 2020). However, the support value was
lower because only the 28S gene was used in the analysis. In
this tree, the new genus was clustered with the Pucciniaceae,
which was represented by 14 genera and separated from every
genus. In addition, the 28S and 5.8S-ITS2 genes of 46 rust taxa
from 13 genera of Pucciniaceae were analyzed, using Dasyspora
guianensis as outgroup taxa (Figure 3). The MP analyses of the
combined 28S and ITS contained 899 characters. Of these, 537
characters were constant, 105 variable characters were parsimony
uninformative, and 257 were parsimony informative. There
were 11 equally most parsimonious trees, with the first tree
(TL = 1,183, CI = 0.496, RI = 0.630, RC = 0.314). The
phylogenetic trees obtained from ML and BI analyses with the
MCMC algorithm coincided with the MP tree shown in Figure 3. The rust fungi on Hamamelidaceae form three separate clades
on one big branch and represent three different species, namely,
Novopuccinia corylopsidis, N. hamamelidis, and N. sycopsis-
sinensis (Figure 3). The type species of Puccina, namely,
P. graminis, was placed within the Puccinia/Uromyces subclade. Novopuccinia was distantly related to the clade represented by
type species P. graminis and closely related to Stereostratum, with
lower support value (Figure 3). Etymology: Novopuccinia (Lat.) referring to the new type taxon
in Puccinia-complex. Descriptions: Spermogonia subepidermal and mostly globose. Aecia subepidermal in origin, cupulate with well-developed
peridium (Aecidium-type). Uredinia subepidermal in origin and
becoming erumpent (Uredo-type). Telia subepidermal in origin,
becoming erumpent, either pulvinate or columnar; teliospores
one-celled and catenulate with a long intercalary cell or two-
celled and pedicellate. Habitat
and
distribution:
Currently
only
known
on
Hamamelidaceae in Asia. Notes: Morphologically, the telia/teliospores of Novopuccinia
consist of two types. One is represented by N. sycopsis-sinensis,
which produces one-celled, catenulate teliospores in columnar
telia. It is taxonomically distinct from other morphologically
similar genera in Pucciniaceae, i.e., Ceratocoma, Chardoniella,
Dietelia, and Trichopsora. Novopuccinia (Figure 4), Chardoniella,
and Trichopsora produce hair-like telial columns and long,
pedicel-like intercalary cells between individual teliospores
(Lagerheim, 1891; Kern, 1939). Morphology Rust sori from fresh collections were free-hand sectioned using a
double-edged blade under a binocular microscope. Spores were
scraped from sori on the dried specimens. Thin sections and
spores were mounted in a drop of lactophenol solution on a
microscope slide and observed, measured, and photographed
using a Leica DM3000 upright microscope (Leica, Germany). For scanning electron microscopy (SEM), fragments of dried
leaves with sori and spores were mounted on SEM stubs with
double-sided tape. The samples were coated with gold using a
Hitachi SCD-005 sputter coater and examined with a Hitachi 3http://mafft.cbrc.jp/alignment/server/
4www.treebase.org April 2021 | Volume 12 | Article 648890 Frontiers in Microbiology | www.frontiersin.org 2 Liu et al. Rust Fungi on Hamamelidaceae FIGURE 1 | (A) Type species location of Novopuccinia on Mt. Sanqingshan of Jiangxi Province, China. (B) The indigenous host, Sycopsis sinensis, occurring to the
mountain. FIGURE 1 | (A) Type species location of Novopuccinia on Mt. Sanqingshan of Jiangxi Province, China. (B) The indigenous host, Sycopsis sinensis, occurring to the
mountain. RESULTS In contrast, Ceratocoma and
Dietelia have small, often inconspicuous, intercalary cells
(Buriticá and Hennen, 1980; Buriticá, 1991; Cummins and
Hiratsuka, 2003). The presence of a well-developed peridium
in the telia is also a distinct characteristic of Novopuccinia,
separating it from Ceratocoma, Chardoniella, and Trichopsora,
which produce non-peridiate telia (Cummins and Hiratsuka,
2003; Berndt, 2018). A metabasidium replaces a probasidium
without
substantial
morphological
change
in
Trichopsora,
while Ceratocoma, Chardoniella, and Novopuccinia produce a
metabasidium external to a probasidium. In addition to these Taxonomy Taxonomy
Novopuccinia Y. M. Liang & Y. Liu, gen. nov. MycoBank MB838356 April 2021 | Volume 12 | Article 648890 Frontiers in Microbiology | www.frontiersin.org 3 Liu et al. Rust Fungi on Hamamelidaceae FIGURE 2 | Pucciniales. Phylogram obtained from maximum parsimony, maximum likelihood, and Bayesian analysis of 28S. The tree is rooted with Eocronartium
muscicola. Genera represented by types are blue; numbers above branches indicate bootstrapping support (1,000 replicates) for each node as MP/ML. Thickened
branches indicate PP > 0.95 from the Bayesian inferences. Bars: 20.0 nucleotide substitutions. The new genus proposed is in boldface. morphological distinctions, N. sycopsis-sinensis with one-celled
teliospores is clearly separated from Ceratocoma, Chardoniella,
and Dietelia in the molecular phylogenetic tree (Figures 2, 3:
molecular data of Trichopsora for the analysis was lacking). morphological distinctions, N. sycopsis-sinensis with one-celled
teliospores is clearly separated from Ceratocoma, Chardoniella,
and Dietelia in the molecular phylogenetic tree (Figures 2, 3:
molecular data of Trichopsora for the analysis was lacking). Another telium/teliospore type is similar to those of Puccinia
in two-celled with pedicellate (Figure 4), but it can be
distinguished from almost all other species by thick-walled
teliospores and Hamamelidaceae host. Our phylogenetic results April 2021 | Volume 12 | Article 648890 Frontiers in Microbiology | www.frontiersin.org 4 Liu et al. Rust Fungi on Hamamelidaceae FIGURE 3 | Phylogenetic tree in Pucciniaceae based on maximum parsimony, maximum likelihood, and Bayesian analysis of the ITS and 28S sequences. Dasyspora
guianensis was used as outgroup. Genera represented by types are blue; numbers above branches indicate bootstrapping support (1,000 replicates) for each node
as MP/ML. Thickened branches indicate PP > 0.95 from the Bayesian inferences. Bars: 30.0 nucleotide substitutions. The new genus proposed is in boldface. FIGURE 3 | Phylogenetic tree in Pucciniaceae based on maximum parsimony, maximum likelihood, and Bayesian analysis of the ITS and 28S sequences. Dasyspora
guianensis was used as outgroup. Genera represented by types are blue; numbers above branches indicate bootstrapping support (1,000 replicates) for each node
as MP/ML. Thickened branches indicate PP > 0.95 from the Bayesian inferences. Bars: 30.0 nucleotide substitutions. The new genus proposed is in boldface. Peridium cells rectangular to rounded, 20–31 × 18–23 µm; the
wall colorless, 3–5 µm thick, and rough-surfaced. also supported the independence of this type of rust on
Hamamelidaceae from the large genus Puccinia (Figures 2, 3). Notes: N. Frontiers in Microbiology | www.frontiersin.org Taxonomy sycopsis-sinensis can be easily distinguished from
other similar taxa by elongated pedicel-like intercalary cells, one-
celled, and peridium cells (Figure 4). According to Tranzschel’s
law, the species has an endocyclic life cycle (Buriticá and Hennen,
1980; Buriticá, 1991). Novopuccinia sycopsis-sinensis Y. Liu & Y. M. Liang, sp. nov. Figure 4 g
MycoBank MB 838357 g
MycoBank MB 838357 Typification: CHINA. Mt. Sanqingshan, Jiangxi Province,
28◦53′88′′N, 118◦04′57′′E, 1,340 m alt., April 1, 2017, on
Sycopsis sinensis Oliv, coll. Y.M. Liang (Holotype: BJFC-R02976;
Paratype: BJFC-R02983). Novopuccinia corylopsidis (Cummins) Y. Liu & Y. M. Liang,
comb. nov. Figure 5 Novopuccinia corylopsidis (Cummins) Y. Liu & Y. M. Liang,
comb. nov. Figure 5 Etymology: Named after the host name of the type species. MycoBank MB 838358 MycoBank MB 838358 Descriptions: Spermogonia, aecia, and uredinia unknown. Telia hypophyllous or petiolicolous, orange, hairlike columnar,
exposed, and 2–5 mm long. Teliospores one-celled with elongated
pedicel-like intercalary cells (25 µm long), ellipsoid to obovoid,
20–35 × 9–14 µm, and attenuate toward both ends; cytoplasm
yellow; the wall colorless, 0.5–1 µm thick, and densely verrucose. Basionym: Puccinia corylopsidis Cummins, Mycologia 43(1):
83 (1951) Descriptions: Spermogonia, aecia, and uredinia unknown. Telia mostly hypophyllous, round, pulvinate, 0.1–0.4 mm in
diameter, and blackish brown. Teliospores variable in shape, wall
thickness, and surface structure, including two types. One kind April 2021 | Volume 12 | Article 648890 Frontiers in Microbiology | www.frontiersin.org 5 Liu et al. Rust Fungi on Hamamelidaceae FIGURE 4 | Morphology of Novopuccinia sycopsis-sinensis (BJFC-R02976, holotype). (A) Sori on a leaf and branches. (B) A teliospore, as seen by SEM. (C) Longitudinal section of a teliospore, as seen by LM. (D) Peridium cells, as seen by LM. (E) Teliospores, as seen by LM. (F) Teliospores, as seen by SEM. (G,H)
Basidia as seen by LM. Scale bars: (B,C) = 1 mm; (D,H) = 10 µm. FIGURE 4 | Morphology of Novopuccinia sycopsis-sinensis (BJFC-R02976, holotype). (A) Sori on a leaf and branches. (B) A teliospore, as seen by SEM. (C) Longitudinal section of a teliospore, as seen by LM. (D) Peridium cells, as seen by LM. (E) Teliospores, as seen by LM. (F) Teliospores, as seen by SEM. (G,H
Basidia as seen by LM. Scale bars: (B,C) = 1 mm; (D,H) = 10 µm. to Novopuccinia, with strong support (Figures 2, 3), hence the
new combination in the new genus. Sycopsis sinensis is the new
host found for N. Frontiers in Microbiology | www.frontiersin.org Taxonomy corylopsidis in this study. was regarded as thin side wall and apex wall slightly thickened
or not thickened, 1–2 µm thick, light yellow, oval, 47–65 × 16–
20 µm, not or hardly constricted at septum, the wall nearly
smooth. The other kind was characterized by the wall apically
and unilaterally thickened, 1.5–9 µm thick at sides, up to 18 µm
thick at apex, mostly fusoid to oval, 46–82 × 16–29 µm, not or
hardly constricted at septum, the wall rugose or densely covered
with irregular verrucous wrinkles, and yellowish to brown; the
pedicel hyaline, thick-walled, persistent, and up to 150 µm. One-
celled spores rarely seen. All these variant spores are produced
in the same sorus. Novopuccinia hamamelidis (Dietel) Y. Liu & Y. M. Liang,
comb. nov. Figure 6 Novopuccinia hamamelidis (Dietel) Y. Liu & Y. M. Liang,
comb. nov. Figure 6 MycoBank MB 838359 Scale bars: (A) = 1 mm; (B,E) = 10 µm. RE 5 | Morphology of Novopuccinia corylopsidis (BJFC-R02977, B–E). (A) Information of type specimen. (B) Sori on a leaf. (C,D) T
ospores, as seen by SEM. Scale bars: (A) = 1 mm; (B,E) = 10 µm. (1989), July 27, on Hamamelis japonica Siebold & Zucc., coll. Y. Ono & H. Miura (BJFC-R3805 = IBAR-3291); Tochigi et al. (1990), August 26, on Hamamelis japonica Siebold & Zucc., coll. M. Sawamura and N. Matsumoto (BJFC-R3806 = IBAR-5475). thickening much paler, smooth, or perhaps minutely rugose
on the broad part of the spore; pedicels mostly not collapsing,
colorless, long and tapering, 250 µm long; one-celled spores rare
(For telia stage, cited from Hiratsuka et al., 1992). Habitat
and
distribution:
On
Hamamelis
japonica
(spermogonial/aecial stages) and Sasamorpha spp. (uredinial/
telial stages) in Japan. Notes: Dietel (1899) described this fungus as Aecidium
hamamelidis based on the Aecidium-type Aecia on Hamamelis
plants. It is also related to the bamboo rust P. mitriformis S. Ito reported by Hiratsuka and Sato (1970). We did not observe
the type specimen of A. hamamelidis, but specimens that were
collected from the same region and host with type collection
were researched. For further study, here we designate BJFC-
R3798 as the epitype specimen. Our molecular phylogenetic
analysis recovered A. hamamelidis within the new genus, with
strong support (Figures 2, 3), hence the new combination in
Novopuccinia. g
p
Specimens examined: JAPAN. Ibaraki et al. (1992), August 10,
on Hamamelis japonica Siebold and Zucc (1992) and Higuchi
et al. (1998) (epitype: BJFC-R3798 = IBAR-6666) and June 28,
(BJFC-R3807 = IBAR-6615); June 7, on Hamamelis japonica
Siebold & Zucc., coll. Y. Ono & K. Ishimiya (BJFC-R3799 = BAR-
8075); Ibaraki et al. (1994), May 13, on Hamamelis japonica
Siebold & Zucc., coll. Y. Ono (BJFC-R3800 = IBAR-7170);
August 13, 1992, on Hamamelis japonica Siebold & Zucc., coll. K. Higuchi, K. Suganuma, and K. Hashimoto (BJFC-R3801 = IBAR-
6553); Aomori, Mt. Hakkodasan, July 31, 1992, on Hamamelis
japonica Siebold & Zucc., Isono (1973) (BJFC-R3802 = IBAR-
5567); Gumma, Tone-gun, June 18, on Hamamelis japonica
Siebold & Zucc., coll. Y. Ono (BJFC-R3803 = IBAR-1585);
Tochigi et al. (1992), July 15, on Hamamelis japonica Siebold &
Zucc., coll. Y. Ono (BJFC-R3804 = IBAR-2194); Ibaraki et al. MycoBank MB 838359 MycoBank MB 838359 Basionym: Aecidium hamamelidis Dietel (1899), Bot. Jb. 27: 571 Synonyms: Puccinia mitriformis Ito (1909), J. Coll. Agric. Tohaku Imper. Univ. 3(2): 233 Aecidium hamamelidis Magnus (1901), Ber. Dt. Bot. Ges. 19: 297 Aecidium hamamelidis Magnus (1901), Ber. Dt. Bot. Ges. 19: 297 Habitat and distribution: On Sycopsis sinensis and Corylopsis
sp. in China (Cummins, 1951; Zhuang et al., 2003). Habitat and distribution: On Sycopsis sinensis and Corylopsis
sp. in China (Cummins, 1951; Zhuang et al., 2003). Descriptions: Spermogonia and aecia occur on species of
Hamamelis; spermogonia abaxial, round, 140 µm in diameter. Aecia hypophyllous, cup shape, the cross-section diameter
is about 140–150 µm. Aeciospores obovoid or globose, 20–
32 × 28–30 µm, surface is covered with irregular verrucosa. Peridial cells round, 30 µm in diameter, outer walls smooth,
inner walls moderately rugose with regular papillae. Telia on
abaxial surface of Sasa plants, early exposed, chocolate-brown;
teliospores 95–130 × 18–22 µm, walls usually unilaterally
thickened, 2–3 µm on the thin side, somewhat to much thicker
on opposite side, 40–70 µm apically, golden brown but the apical Specimens examined: China. Lao Shan et al., 1931, April
30, on Corylopsis sp. (type: BPI 058871); Sanqingshan, Jiangxi
Province, 28◦53′95′′N, 118◦04′52′′E, 1,342 m alt. (BJFC-R02977)
and 28◦53′88′′N, 118◦04′57′′, 1,340 m alt. (BJFC-R02978), April
1, 2017, on Sycopsis sinensis Oliv., coll. Y.M. Liang. Notes: The species was first reported on Corylopsis plants
and named P. corylopsidis based on Puccinia-like teliospores
(Cummins, 1951). However, it differs from other Puccinia
species by dimorphic teliospores (Figure 5). Our molecular
phylogenetic analysis indicated that P. corylopsidis closely related April 2021 | Volume 12 | Article 648890 Frontiers in Microbiology | www.frontiersin.org 6 Liu et al. Rust Fungi on Hamamelidaceae FIGURE 5 | Morphology of Novopuccinia corylopsidis (BJFC-R02977, B–E). (A) Information of type specimen. (B) Sori on a leaf. (C,D) Teliospores, as seen by LM. (E) Teliospores, as seen by SEM. Scale bars: (A) = 1 mm; (B,E) = 10 µm. FIGURE 5 | Morphology of Novopuccinia corylopsidis (BJFC-R02977, B–E). (A) Information of type specimen. (B) Sori on a leaf. (C,D) Teliospores, as seen by LM. (E) Teliospores, as seen by SEM. Scale bars: (A) = 1 mm; (B,E) = 10 µm. FIGURE 5 | Morphology of Novopuccinia corylopsidis (BJFC-R02977, B–E). (A) Information of type specimen. (B) Sori on a leaf. (C,D) Teliospores, as seen by LM. (E) Teliospores, as seen by SEM. Frontiers in Microbiology | www.frontiersin.org DISCUSSION No molecular data have been used to analyze
the relationship between these two species and Novopuccinia
on Hamamelidaceae, although we may assume that they are
closely related to N. corylopsidis and N. hamamelidis based on the
observation of similar teliospores, hosts of spermogonial/aecial
stages, and distribution. Further investigations will confirm
this conjecture. The new genus is sister to Stereostratum (Figure 3), but
they differ in teliospore characters. Novopuccinia contains two
teliospore types (Figures 4, 5), while Stereostratum has one
kind of teliospore (Mangus, 1898). Teliospores of N. corylopsidis
and N. hamamelidis are thick-walled and have one germ pore
in each cell (Figure 5; Hiratsuka and Sato, 1970; Hiratsuka
et al., 1992), while Stereostratum spp. have three germ pores
in each teliospore cell, and their wall does not thicken
(Mangus, 1898). The type species of Novopuccinia, namely, N. sycopsis-sinensis,
is microcyclic in the life cycle and produces catenulate, rough-
surfaced teliospores with intercalary cells between them. The
similar life cycle pattern and morphological characteristics
and close phylogenetic relationship to N. corylopsidis and
N. hamamelidis (Figures 2, 3) indicate that N. sycopsis-
sinensis was derived from a hypothetical puccinioid ancestor
by reduction of the life cycle through the endophylloid
pathway (Jackson, 1931; Buriticá and Hennen, 1980). In this
pathway, morphologically Aecidium-type telia of a derived
microcyclic species are produced on a spermogonial/aecial
host plant of a putative ancestral macrocyclic heteroecious
species. Similarly, N. corylopsidis might have been derived
from the same hypothetical ancestor by reduction of the life
cycle through the telioid pathway (Jackson, 1931; Buriticá
and Hennen, 1980). The latter evolutionary pathway is well-
known and is accounted for by Tranzschel’s Law (Shattock
and
Preece,
2000). These
two
evolutionary
phenomena
and resulting correlations between macrocyclic heteroecious
species and microcyclic species have been corroborated to
a slight extent by a molecular phylogenetic method. Early
successful studies include graminicolous puccinias (Zambino
and Szabo, 1993), macrocyclic heteroecious Cronarium and
endocyclic
Peridermium
(Vogler
and
Bruns,
1998),
the
P. monoica species complex (parasitizing on Brassicaceae and
Poaceae) (Roy et al., 1998), and the P. hemerocallidis species DISCUSSION Consequently, several genera have diverged from the
genus Puccinia sensu lato, despite their similarity in two-celled
teliospore morphology such as Allodus (Minnis et al., 2012),
Austropuccinia (Beenken, 2017), and Puccorchidium (Beenken
and Wood, 2015). However, genera that have been separated from
Puccinia based on differences in their morphology appear within
Puccinia, such as Uromyces, Diorchidium, Cumminsiella, and
Miyagia (Figures 2, 3; Aime, 2006; Van Der Merwe et al., 2008;
Beenken and Wood, 2015). New combined species, Novopuccinia
corylopsidis and N. hamamelidis, were defined by combining
their distant relationship to type species of Puccinia clade,
their close relationship with type species N. sycopsis-sinensis
(Figures 2, 3), common hosts, and evolution hypothesis of the
endocyclic life cycle. complex (parasitizing Caprifoliaceae and Xanthorrhoeaceae)
(Chatasiri et al., 2006). Sycopsis sinensis (Hamamelidaceae) has been listed as a
protected species in China based on its ecological value and
rarity (Zang and Bian, 2003; Xu et al., 2007). No disease
has been reported in this species to date. Two distinct
rust fungi on two adjacent S. sinensis trees, N. sycopsis-
sinensis and N. corylopsidis, were found in the present study. N. corylopsidis has earlier been reported on Corylopsis sp. (Cummins, 1951), and now we observe host expansion and
parasitism involving S. sinensis. At the same time, we suspect
that the species had mutated to adapt to the environment,
thus
forming
a
new
rust
N. sycopsis-sinensis
that
also
causes disease. In addition to these three species, two more species have
been reported on Hamamelidaceae in Japan, namely, P. sasicola
(Hara) Hino & Katumoto on C. glabrescens (spermogonial/aecial
stages) and Sasamorpha borealis (telial stage), P. sakamotoi
Hirats. f. & Yoshin on Distylium spp. (Hiratsuka, 1942; Hino and
Katumoto, 1955). The former is heteroecious and demicyclic; its
teliospores are thick-walled, similar to those of N. corylopsidis
and N. hamamelidis (Hino and Katumoto, 1955; Hiratsuka
et al., 1992). However, it differs from the latter two species
by its demicyclic life cycle and hosts during telia stage. The
latter is demicyclic and autoecious on Distylium, cylindrical telia
are similar to N. sycopsis-sinensis (Figure 4), and two-celled
teliospores with thick walls are similar to N. corylopsidis and
N. hamamelidis (Figure 5; Hiratsuka et al., 1992). However,
teliospores of the latter are one-celled with elongated pedicel-
like intercalary cells, while P. sakamotoi produces two-celled
teliospores without intercalary cells (Hiratsuka, 1942). An
autoecious and demicyclic life cycle can also distinguish it from
similar species. DISCUSSION In this study, three rust taxa obtained from Hamamelidaceae were
identified based on morphologic characters and phylogeny based
on the 28S and 5.8S-ITS2 sequences. As a result, Novopuccinia April 2021 | Volume 12 | Article 648890 Frontiers in Microbiology | www.frontiersin.org 7 Liu et al. Rust Fungi on Hamamelidaceae FIGURE 6 | Morphology of Novopuccinia hamamelidis BJFC-R3798 (IBAR6666). (A) Aecia on a leaf. (B) Spermogonia on a leaf. (C) Aeciospores, as seen by LM. (D,E) Aeciospores, as seen by SEM. (F,G) Peridial cells, as seen by SEM. Scale bars: (A,B) = 100 mm; (C,G) = 10 µm. FIGURE 6 | Morphology of Novopuccinia hamamelidis BJFC-R3798 (IBAR6666). (A) Aecia on a leaf. (B) Spermogonia on a leaf. (C) Aeciospores, as seen by LM. (D,E) Aeciospores, as seen by SEM. (F,G) Peridial cells, as seen by SEM. Scale bars: (A,B) = 100 mm; (C,G) = 10 µm. are verrucose and catenulate with intercalary cells. These
taxa are considered to have been derived from a Puccinia–
Uromyces complex by reduction of the life cycle through the
endophylloid pathway; their occurrence on the same host plants
may nonetheless provide clues on the relationship between
microcyclic and long-cycled autoecious rust fungi (Jackson,
1931; Buriticá and Hennen, 1980). Members of this complex
are closely related to the environment and occur mainly in
tropical and other warm regions (Berndt, 2018). The second typified with N. sycopsis-sinensis is proposed as a new genus
in Pucciniaceae. The two known species, P. corylopsidis and
Aecidium hamamelidis, were combined to the new genus
(N. corylopsidis and N. hamamelidis) by the same host plant, the
molecular data, and endocyclic life cycle. Novopuccinia includes two different teliospore types. The
first type is represented by type species N. sycopsis-sinensis
with one-celled teliospores and belongs to the kind of taxa,
whose telia are cupulate, usually with peridia, and teliospores April 2021 | Volume 12 | Article 648890 Frontiers in Microbiology | www.frontiersin.org 8 Rust Fungi on Hamamelidaceae Liu et al. type is similar to Puccinia-like teliospores and represented by
P. corylopsidis and Aecidium hamamelidis (synonym: Puccinia
mitriformis), but phylogenetically closely related to Novopuccinia
sycopsis-sinensis (Figures 2, 3). While teliospore characters
are deemed important to distinguishing genera and families
within Pucciniales (Cummins and Hiratsuka, 2003), molecular
phylogenies show that their morphological characteristics may
not always define higher taxa (Aime, 2006; Beenken and Wood,
2015). Frontiers in Microbiology | www.frontiersin.org REFERENCES Hara, K. (1939). On the life-history of Puccinia sasaecola Hara on Sasamorpha
purpurescens Nakai. Bot. Zool. 7, 1618–1620. Aime, M. C. (2006). Toward resolving family-level relationships in rust
fungi (Uredinales). Mycoscience 47, 112–122. doi: 10.1007/s10267-006-
0281-0 p
p
Hara, K. (1952). Inoculation experiments with Puccinia sasaecola Hara. Ann. Phytopathol. Soc. Jpn. 16:71. doi: 10.3186/jjphytopath.16.71 p
Phytopathol. Soc. Jpn. 16:71. doi: 10.3186/jjphytopath.16.71 Hino, I., and Katumoto, K. (1955). Illustrationes fungorum bambusicolorum III. Bull. Fac. Agr. Yamaguti Univ. 6, 29–68. Aime, M. C., and Mctaggart, A. (2020). A higher-rank classification for rust
fungi, with notes on genera. Fungal Syst. Evol. 7, 21–47. doi: 10.3114/fuse.2021. 07.02 Hiratsuka, N. (1942). Notae Uredinologiae Asiae Orientalis. I. Bot. Mag. 1, 374–
382. doi: 10.15281/jplantres1887.56.374 Beenken, L. (2017). Austropuccinia: a new genus name for the myrtle rust Puccinia
psidii placed within the redefined family Sphaerophragmiaceae (Pucciniales). Phytotaxa 297, 53–61. doi: 10.11646/phytotaxa.297.1.5 Hiratsuka, N., and Sato, S. (1970). Notes on Puccinia mitriformis S. Ito. Reps. Tottori Mycol. Inst. 8, 39–44. Hiratsuka, N., Sato, S., Katsuya, K., Kakishima, M., Hiratsuka, Y Hiratsuka, N., Sato, S., Katsuya, K., Kakishima, M., Hiratsuka, Y., Kaneko, S., et al. (1992). The Rust Flora of Japan. Tsukuba: Tsukuba Shuppankai. Beenken, L., and Wood, A. R. (2015). Puccorchidium and Sphenorchidium, two new
genera of Pucciniales on Annonaceae related to Pucciniapsidii and the genus
Dasyspora. Mycol. Prog. 14, 1–13. doi: 10.1007/s11557-015-1073-8 Ito, K. (1909). On the Uredinales parasitic on the Japanese Gramineae. J. Coll. Agr. Tohoku Imp. Univ. 3, 180–265. Jackson, H. S. (1931). Present evolutionary tendencies and the origin of life cycles
in the Uredinales. Mem. Torrey Bot. Club 18, 5–108. Beenken, L., Zoller, S., and Berndt, R. (2012). Rust fungi on Annonaceae II: the
genus Dasyspora Berk. & M.A. Curtis. Mycologia 104, 659–681. doi: 10.3852/
11-068 Katoh, K., and Standley, D. M. (2013). MAFFT multiple sequence alignment
software version 7: improvements in performance and usability. Mol. Biol. Evol. 30, 772–780. doi: 10.1093/molbev/mst010 Berndt, R. (2018). The Pucciniosiraceae: taxonomy of a polyphyletic family of the
rust fungi (Uredinales). Biosyst. Ecol. Ser. 34, 245–269. Buriticá, P., and Hennen, J. F. (1980). Pucciniosireae (Uredinales, Pucciniaceae). Flora Neotrópica, 24. New York, NY: New York Botanical Garden. Kern, F. D. (1939). Chardoniella—a new genus of the Uredinales. Mycologia 31,
373–375. doi: 10.1080/00275514.1939.12017352 Buriticá, P. C. (1991). Familias del orden Uredinales con ciclo de vida
completamente reducido. Revta Acad Colomb Cienc Exact Fís Nat. 18, 131–148. Lagerheim, G. (1891). ACKNOWLEDGMENTS We
thank
Prof. M. Kakishima,
University
of
Tsukuba,
Tsukuba, Japan, for providing comments relating to this
report. We express our gratitude to Dr. Lisa A. Castlebury
(USDA ARS Mycology and Nematology Genetic Diversity and
Biology Laboratory, Baltimore, United States) for providing
us with photomicrographs of Puccinia corylopsidis for use in
this study. DATA AVAILABILITY STATEMENT The
datasets
presented
in
this
study
can
be
found
in
online repositories. The names of the repository/repositories
and accession number(s) can be found below: https://www. ncbi.nlm.nih.gov/genbank/ MN943642, MN943643, MW136693,
MW136697, MW386782, MW386783, MW394517. FUNDING The National Natural Science Foundation Program of China (No. 31870628) supported this study. The National Natural Science Foundation Program of China (No. 31870628) supported this study. AUTHOR CONTRIBUTIONS The Supplementary Material for this article can be found
online
at:
https://www.frontiersin.org/articles/10.3389/fmicb. 2021.648890/full#supplementary-material YLa and CT conceived and designed the experiments. YLu,
BC, and WL performed the experiments. YLu and YO analyzed KEY TO RUST FUNGAL SPECIES THAT
WERE RECORDED ON THE HOST
FAMILY HAMAMELIDACEAE IN ASIA 1. Life cycle macrocyclic, heteroecious .......................................... 2
1. Life cycle microcyclic or demicyclic and autoecious ............... 3 1. Life cycle microcyclic or demicyclic and autoecious ............... 3
2. Teliospores
two-celled,
pedicellate,
wall
apically
prominently elongated; spermogonial and aecial stages
on
Corylopsis
(Trib. Corylopsideae);
distributed
in
Japan .............................................................. Puccinia sasicola
2. Teliospores
two-celled,
pedicellate,
wall
mostly
unilaterally thickened; spermogonial and aecial stages
on Hamamelis (Trib. Hamamelideae); distributed in
Japan ............................................ Novopuccinia hamamelidis 2. Teliospores
two-celled,
pedicellate,
wall
apically
prominently elongated; spermogonial and aecial stages
on
Corylopsis
(Trib. Corylopsideae);
distributed
in
Japan .............................................................. Puccinia sasicola 2. Teliospores
two-celled,
pedicellate,
wall
apically
prominently elongated; spermogonial and aecial stages
on
Corylopsis
(Trib. Corylopsideae);
distributed
in
Japan .............................................................. Puccinia sasicola p
2. Teliospores
two-celled,
pedicellate,
wall
mostly
unilaterally thickened; spermogonial and aecial stages
on Hamamelis (Trib. Hamamelideae); distributed in
Japan ............................................ Novopuccinia hamamelidis April 2021 | Volume 12 | Article 648890 Frontiers in Microbiology | www.frontiersin.org 9 Rust Fungi on Hamamelidaceae Liu et al. 3. Telia columnar; teliospore one-celled, catenulate; only telia
produced on Sycopsis (Trib. Fothergilleae); distributed in
China ........................................... Novopuccinia sycopsis-sinensis
3. Telia pulvinate .............................................................................. 4
4. Teliospores two-celled, pedicellate, wall mostly unilaterally
thickened;
only
telia
produced
on
Corylopsis
(Trib. Corylopsideae) and Sycopsis (Trib. Distylteae); distributed in
China ................................................... Novopuccinia corylopsidis
4. Teliospores two-celled, pedicellate, wall evenly thickened
at sides; aecia and telia produced on Distylium (Trib. Fothergilleae); distributed in Japan ............................ Puccinia
sakamotoi 3. Telia columnar; teliospore one-celled, catenulate; only telia
produced on Sycopsis (Trib. Fothergilleae); distributed in
China ........................................... Novopuccinia sycopsis-sinensis
3 Telia pulvinate
4 the data. YLu wrote the manuscript. YO and YLa revised
and approved the final version of the manuscript. All authors
contributed extensively to the work presented in this manuscript. 3. Telia columnar; teliospore one-celled, catenulate; only telia
produced on Sycopsis (Trib. Fothergilleae); distributed in
China ........................................... Novopuccinia sycopsis-sinensis
3. Telia pulvinate .............................................................................. 4
4. Teliospores two-celled, pedicellate, wall mostly unilaterally
thickened;
only
telia
produced
on
Corylopsis
(Trib. Corylopsideae) and Sycopsis (Trib. Distylteae); distributed in
China ................................................... Novopuccinia corylopsidis
4. Teliospores two-celled, pedicellate, wall evenly thickened
at sides; aecia and telia produced on Distylium (Trib. Fothergilleae); distributed in Japan ............................ Puccinia
sakamotoi REFERENCES Pucciniosira, Chrysopsora, Alveolaria und Trichopsora, vier
neue Uredineen-Gattungen mit tremelloider Entwickelung. Ber. Deutsch. Bot. Ges. 9, 344–348. Chatasiri, S., Kitade, O., and Ono, Y. (2006). Phylogenetic relationships among
Puccinia hemerocallidis, P. funkiae, and P. patriniae (Uredinales) inferred from
ITS sequence data. Mycoscience 47, 123–129. doi: 10.1007/S10267-006-0283-Y Liu, Y., Cao, B., Tao, S. Q., Tian, C. M., and Liang, Y. M. (2018). Phragmidium
species parasitizing species of Rosaceae inTibet, China, with descriptions
of three new species. Mycol. Prog. 17, 967–988. doi: 10.1007/s11557-018-
1406-5 Cummins, G. B. (1951). Uredinales from continental China collected by S.Y. Cheo. II. Mycologia 43, 78–98. doi: 10.2307/3755549 Cummins, G. B., and Hiratsuka, Y. (2003). Illustrated Genera Of Rust Fungi III. Saint Paul, MN: American Phytopathological Society. Magnus, P. (1901). Ueber einige von J. Bornmüller im Jahre 1900 auf
den canarischen Inseln gesammelte Uredineen. Ber. Dtsch. Bot. Ges. 19,
292–300. Darriba, D., Taboada, G. L., Doallo, R., and Posada, D. (2012). jModelTest 2:
more models, new heuristics and parallel computing. Nat. Methods 9:772. doi:
10.1038/nmeth.2109 Minnis, A. M., McTaggart, A. R., Rossman, A. Y., and Aime, M. C. (2012). Taxonomy of mayapple rust: the genus Allodus resurrected. Mycologia 104,
942–950. doi: 10.3852/11-350 Dietel, P. (1899). Uredineae japonicae I. Bot. Jahrb. Syst. 27, 564–576. Frontiers in Microbiology | www.frontiersin.org April 2021 | Volume 12 | Article 648890 10 Liu et al. Rust Fungi on Hamamelidaceae O’Donnell, K. (1993). “Fusarium and its near relatives,” in The fungal Holomorph:
Mitotic, Meiotic and Pleomophic Speciation in Fungal Systematics, eds D. R. Reynolds and J. W. Taylor (Wallingford: CABI), 225–233. Vogler, D. R., and Bruns, T. D. (1998). Phylogenetic relationships among the pine
stem rust fungi (Cronartium and Peridermium spp.). Mycologia 90, 244–257. doi: 10.1080/00275514.1998.12026904 Xu, K. Y. (2001). Preliminary Ecology Study on Sycopsis sinensis community in
Honhe National Nature Reserve, Hubei Province. Dissertation, Hubei, Central
China Normal University. Ono, Y. (2017). Rust fungi (Pucciniales) in northern Ibaraki prefecture. Bull. Coll. Educ. Ibaraki Univ (Nat. Sci) 66, 37–56. Posada, D., and Crandall, K. A. (1998). Modeltest: testing the model of DNA
substitution. Bioinformatics 14, 817–818. doi: 10.1093/bioinformatics/14.9.817 Xu, S. D., Han, Q. Y., Li, D. X., Guo, Y. D., Li, G., and Li, P. (2007). A large area of
Sycopsis sinensis communities were found in the Dalaoling nature reserve of the
three gorges. Hubei Forestry Sci. Technol. 4, 61–68. Rannala, B., and Yang, Z. (1996). REFERENCES Probability distribution of molecular evolutionary
trees: a new method of phylogenetic inference. J. Molec. Evol. 43, 304–311. doi: 10.1007/BF02338839 three gorges. Hubei Forestry Sci. Technol. 4, 61–68. Zambino, P. J., and Szabo, L. J. (1993). Phylogenetic relationships of selected cereal
and grass rusts based on rDNA sequence analysis. Mycologia 85, 401–414. doi: 10.2307/3760702 Ronquist, F., and Huelsenbeck, J. P. (2003). MrBayes 3: bayesian phylogenetic
inference under mixed models. Bioinformatics 19, 1572–1574. doi: 10.1093/
bioinformatics/btg180 Zang, M., and Bian, X. M. (2003). A study on the rare and threatened plants in Mt. Sanqingshan, Jiangxi Province. J. Wuhan Bot. Res. 21, 515–520. Roy, B. A., Vogler, D. R., Burns, T. D., and Szaro, T. M. (1998). Cryptic species
in the Puccinia monoica complex. Mycologia 90, 846–853. doi: 10.2307/37
61326 Zhuang, J. Y., Wei, S. X., and Wang, Y. C. (2003). Flora Fungorum Sinicorum. Uredinales II, Vol. 19. Beijing: Science Press. Shattock, R. C., and Preece, T. F. (2000). Tranzschel revisited: modern studies of
the relatedness of different rust fungi confirm his law. Mycologist 14, 113–117. doi: 10.1016/S0269-915X(00)80086-5 Conflict of Interest: The authors declare that the research was conducted in the
absence of any commercial or financial relationships that could be construed as a
potential conflict of interest. Swofford,
D. L. (2003). PAUP:
Phylogenetic
Analysis
Using
Parsimony,
and
Other
Methods,
Version
4.0b10. Sunderland, MA, USA: Sinauer
Associates. The reviewer PZ declared a shared affiliation with one of the authors BC to the
handling editor at the time of review. Tian, C. M., Shang, Y. Z., Zhuang, J. Y., Wang, Q., and Kakishima, M. (2004). Morphological and molecular phylogenetic analysis of Melampsora
species on poplars in China. Mycoscience 45, 56–66. doi: 10.1007/S10267-003-
0150-Z Copyright © 2021 Liu, Cao, Tian, Ono, Lin and Liang. This is an open-access article
distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the
original author(s) and the copyright owner(s) are credited and that the original
publication in this journal is cited, in accordance with accepted academic practice. No
use, distribution or reproduction is permitted which does not comply with these terms. Van Der Merwe, M. M., Walker, J., Ericson, L., and Burdon, J. J. (2008). Coevolution with higher taxonomic host groups within the Puccinia / Uromyces
rust lineage obscured by host jumps. Mycol. Res. 112, 1387–1408. Frontiers in Microbiology | www.frontiersin.org REFERENCES doi: 10.1016/
j.mycres.2008.06.027 April 2021 | Volume 12 | Article 648890 Frontiers in Microbiology | www.frontiersin.org 11
|
https://openalex.org/W2147926439
|
https://thericejournal.springeropen.com/counter/pdf/10.1186/1939-8433-5-23
|
English
| null |
An in planta, Agrobacterium-mediated transient gene expression method for inducing gene silencing in rice (Oryza sativa L.) leaves
|
Rice
| 2,012
|
cc-by
| 10,584
|
Abstract Background: Localized introduction and transient expression of T-DNA constructs mediated by agro-infiltration of
leaf tissues has been largely used in dicot plants for analyzing the transitivity and the cell-to cell movement of the
RNAi signal. In cereals, however, the morphology of the leaf and particularly the structure of the leaf epidermis,
prevent infiltration of a bacterial suspension in cells by simple pressure, a method otherwise successful in dicots
leaves. This study aimed at establishing a rapid method for the functional analysis of rice genes based on the
triggering of RNA interference (RNAi) following Agrobacterium-mediated transient transformation of leaves. Background: Localized introduction and transient expression of T-DNA constructs mediated by agro-infiltration of
leaf tissues has been largely used in dicot plants for analyzing the transitivity and the cell-to cell movement of the
RNAi signal. In cereals, however, the morphology of the leaf and particularly the structure of the leaf epidermis,
prevent infiltration of a bacterial suspension in cells by simple pressure, a method otherwise successful in dicots
leaves. This study aimed at establishing a rapid method for the functional analysis of rice genes based on the
triggering of RNA interference (RNAi) following Agrobacterium-mediated transient transformation of leaves. Results: Using an agro-infection protocol combining a wound treatment and a surfactant, we were able to obtain
in a reliable manner transient expression of a T-DNA-borne uidA gene in leaf cells of japonica and indica rice
cultivars. Using this protocol to transiently inhibit gene expression in leaf cells, we introduced hairpin RNA (hpRNA)
T-DNA constructs containing gene specific tags of the phytoene desaturase (OsPDS) and of the SLENDER 1 (OsSLR1)
genes previously proven to trigger RNAi of target genes in stable transformants. SiRNA accumulation was observed
in the agro-infected leaf area for both constructs indicating successful triggering of the silencing signal. Results: Using an agro-infection protocol combining a wound treatment and a surfactant, we were able to obtain
in a reliable manner transient expression of a T-DNA-borne uidA gene in leaf cells of japonica and indica rice
cultivars. Using this protocol to transiently inhibit gene expression in leaf cells, we introduced hairpin RNA (hpRNA)
T-DNA constructs containing gene specific tags of the phytoene desaturase (OsPDS) and of the SLENDER 1 (OsSLR1)
genes previously proven to trigger RNAi of target genes in stable transformants. SiRNA accumulation was observed
in the agro-infected leaf area for both constructs indicating successful triggering of the silencing signal. Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 An in planta, Agrobacterium-mediated transient
gene expression method for inducing gene
silencing in rice (Oryza sativa L.) leaves An in planta, Agrobacterium-mediated transient
gene expression method for inducing gene
silencing in rice (Oryza sativa L.) leaves Aurélie Andrieu1, Jean Christophe Breitler1, Christelle Siré1, Donaldo Meynard1, Pascal Gantet2
and Emmanuel Guiderdoni1* Aurélie Andrieu1, Jean Christophe Breitler1, Christelle Siré1, Donaldo Meynard1, Pascal Gantet2
and Emmanuel Guiderdoni1* © 2012 Andrieu et al.; licensee Springer. This is an Open Access article distributed under the terms of the Creative Commons
Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction
in any medium, provided the original work is properly cited. RESEARCH Open Access Open Access * Correspondence: emmanuel.guiderdoni@cirad.fr
1CIRAD, UMR AGAP, TAA108/03, Av Agropolis, F-34398 Montpellier Cedex 05,
France
Full list of author information is available at the end of the article
© 2012 Andrieu et al.; licensee Springer. This is an Open Access article distributed under the terms of the Creative Commons
Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction
in any medium, provided the original work is properly cited. Abstract Accumulation of secondary siRNA was observed in both stably and transiently transformed leaf tissues expressing
the HpRNA OsSLR1 construct. Gene silencing signalling was investigated in monitoring the parallel time course of
OsPDS-derived mRNA and siRNA accumulation in the agro-infiltrated leaf area and adjacent systemic sectors. The
sensitive RT-Q-PCR method evidenced a consistent, parallel decrease of OsPDS transcripts in both the agroinfiltred
and adjacent tissues, with a time lag for the latter. Conclusions: These results indicate that the method is efficient at inducing gene silencing in the agro-infected leaf
area. The transfer of low amounts of siRNA, probably occurring passively through the symplastic pathway from the
agro-infected area, seemed sufficient to trigger degradation of target transcripts in the adjacent tissues. This
method is therefore well suited to study the cell-to-cell movement of the silencing signal in a monocot plant and
further test the functionality of natural and artificial miRNA expression constructs. Keywords: Agrobacterium tumefaciens, Gene silencing, Leaf agroinfection, OsPDS, OsSRL1, Rice, Transitivity,
Cross silencing Background can be amplified by a phenomenon called transitivity
which increases the initial pool of siRNAs by producing
secondary siRNAs corresponding to sequences located
outside the primary targeted regions of a transcript
(Nishikura 2001; Sijen et al., 2001). Himber et al. (2003)
have proposed a model of RNA silencing cell-to-cell
movement including a central role for transitivity. The
vast majority of experimental data used to formulate this
model of RNA silencing movement over cells and orga-
nism was accumulated in N. benthamiana and A. tha-
liana. Regarding transitivity of RNA
silencing and
signalling,
experience
on
monocotyledonous
plants
remains fairly limited since the report of the conserva-
tion of the transitive RNA silencing machinery in rice by
Miki et al. (2005). In dicots plants, agro-infection methods allow efficient
transfer and transient expression of T-DNA vectors in
leaf cells. Transient assays by agro-infiltration have thus
been increasingly used as a simple and rapid method for
assaying gene function (Wroblewski et al. 2005; Small
2007). Because they provide a rapid, versatile and con-
venient way for achieving a very high level of gene ex-
pression in a distinct and defined zone of the leaf, these
Agrobacterium-mediated transient expression systems
have been largely used for analyzing the induction of
the RNA silencing process, thereby validating gene
function through down regulation of gene expression -
as illustrated recently in grapevine (Bertazzon et al. 2012)- and analyzing the mechanism of transitivity and
movement of RNA silencing in leaves (e.g. Schöb et al. 1997, Voinnet 2001). The goal of this study was first to establish a standar-
dized Agrobacterium transfection protocol for inducing
transient gene expression in rice leaf cells. The second
objective was to illustrate this method in inducing the si-
lencing of two endogenous rice genes (OsPDS and
OsSRL1) to investigate the transitivity and systemic
spreading of the RNAi signal in a monocotyledonous
leaf. In rice, as in other cereals, the establishment of such a
system is hindered by the monocot leaf structure which
includes the presence of an epidermal cuticular wax and
high silicium content preventing infiltration of bacterial
suspension by simple pressure. Microprojectile bom-
bardment has been used as an alternative delivery
method to introduce expression construct in cereal leaf
cells. Plant Material and in planta agroinfection
h
f
(
) Plant Material and in planta agroinfection
Three varieties of rice (Oryza sativa L.) were used in this
study: (i) the sub tropical japonica rice cv. Zhong
Zuo321 from China, (ii) the temperate japonica rice cv. Nipponbare and (iii) the high value indica rice cv. IR64
from IRRI. Bacteria used for rice agroinfection were pla-
ted on solid AB medium (Chilton et al. 1974) containing
50 mg/l kanamycin sulfate and 75 mg/l rifampicin and
incubated at 28°C for 3 days. The bacteria were then col-
lected with a flat spatula and re-suspended in liquid
agroinfection
medium
(R2
Basic
(Sallaud
et
al. 2003) + 10 g/l glucose, 200 μM acetosyringone, 0,01%
Silwet L-77, pH5.5) by vortexing to give an absorbance
at 600 nm between 0.5 and 0.8. The antepenultimate
emerged leaf of plantlets at tillering stage were mecha-
nically wounded with multiple needles and immersed in
the Agrobacterium suspension at 20°C for 30 to 60 min. The plantlets were grown for 2 to 3 days at 20°C and
then placed under containment greenhouse growth con-
ditions. Agrobacterium-mediated transformation of seed
embryo-derived
embryogenic
callus
was
performed
according to Sallaud et al. 2003. Once established and among a range of applications,
such a method could be used for triggering a localized,
hpRNA-mediated silencing process and further study
the transitivity and spreading of the silencing signal
throughout the whole plant. RNA silencing is non-cell
autonomous and operates through different pathways in-
volving separate mechanisms and, probably, distinct sig-
nals (Voinnet 2005a; Brodersen and Voinnet 2006;
Ghildiyal and Zamore 2009). The systemic silencing sig-
nal reflects with many evidences an antiviral defence
mechanism in plants: non-cell autonomous silencing re-
lies on a systemic signal that moves ahead or follows
RNA viruses by the same way of propagation to initiate
RNA silencing and prevent or delay viral infection
(Voinnet 2005b). While RNA-silencing induction and
RNA degradation have been studied in detail, much less
is known about why and how RNA silencing moves from
cell-to-cell and sometimes spreads systemically in plants. The answer to these questions seems to be closely
related to the transitive RNA silencing observed in
C. elegans and plants. In these organisms, RNA silencing Gene specific tag (GST) and T-DNA constructs
The binary vector pC5300, a pCAMBIA 1300 derivative,
had been described elsewhere (Breitler et al. 2004). Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 Page 2 of 12 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 Background As to gene silencing,
bombardment-mediated
introduction of dsRNA corresponding to endogenous
genes or transgenes in cells of maize, barley and wheat
leaves proved to induce interference with gene function
but restricted to the single cell where the delivery is
achieved (Schweizer et al. 2000). This stresses the need
to establish a rapid, versatile and convenient protocol for
achieving a very high level of gene expression in planta -
e.g. in a distinct and defined zone of the cereal leaf. Real-time PCR analysis Total RNA extraction was performed with Tri-Reagent
(Sigma) according to the manufacturer’s instructions. The template DNA was removed by treatment with the
DNA RQ1 RNase-free DNase kit (Promega). Each cDNA
sample was subjected to real-time PCR analysis in trip-
licate. To
normalize
the
variance
among
samples,
OsExp (Os06g11070.1) was used as endogenous control
(Caldana et al. 2007). Relative expression values were
calculated
after
normalizing
against
the
control
cDNA. Primers were designed from 3' end of the
gene using Primer3plus (http://www.bioinformatics.nl/
cgi-bin/primer3plus/primer3plus.cgi) with QPCR para-
meters ‘on’. Each primer pair was checked for their spe-
cificity using the Primer Blaster tool in OryGenesDB
(http://orygenesdb.cirad.fr/tools.html), which were further
confirmed by dissociation curve analysis obtained after Plant Material and in planta agroinfection
h
f
(
) The
reporter plasmid pC5300-UGN bears the uidA gene
under the control of the entire 5’ untranslated region of
the maize polyubiquitin gene Ubi-1 (Christensen and Page 3 of 12 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 (Amersham). The GST probe was PCR products similar
to GST cloned in the pBIOS738 binary vector. The
5’ probes were produced by PCR using the following
set
of
primers:
5’pds,
5’-GGTGCTTCGCAAGTAG
CAGC-3’ and 5’-TACTAAGAAACAATGAAATG-3’ and
5’slr1,
5’-GTGCAAGGACAAGGTGATGG-3’
and
5’-
GCACGCCCACTTCTACGAGT-3’. The 3’ probes were
produced by PCR with the following set of primers used
for RT-PCR analysis 3’-pds, 5’-TTGTGCTCAGTCTG
TAGTGGA-3’ and 5’-TGTGAAGGGATTAAGAGACC-3’
and 3’-slr1, 5’-GGCACAATTGAAGCTTGACG-3’ and
5’-ATAGATGGGCTAGGAGGACCAAG-3’. Following
hybridisation, the
membrane was washed twice for
20 min at 50°C (2x SSC and 2% SDS) and all hybridisa-
tion signals were detected by phosphorimaging (Storm
820, Amersham). Quail 1996) and the polyadenylation sequence from A. tumefaciens nos gene. The hpRNA binary vector pBIOS
738, kindly provided by W. Paul and P. Perez (BIO-
GEMMA, Clermont-Ferrand, France), was constructed
by cloning the intron of rice tubulin1 gene between twin
GatewayTM cassettes in inverted orientation into a
pCAMBIA 2300 binary vector. This hpRNA expression
cassette is under the control of the cassava vein mosaic
virus (CsVMV) promoter (Verdaguer et al. 1998) and
the polyadenylation sequence from A. tumefaciens nos
gene (Figure 1). For the rice PDS gene a 400 bp fragment (Accession
no. AF049356.1, nucleotides no. 660–1040) was used
as GST (forward primer: 5’-GAAGTTTGCTCTTG
GACTTTTGCCA-3’ and reverse primer: 5’-TATTT
GAGTTCCATCGGTAAGTGCA-3’). For the SLR1 gene (Accession no. AB030956), the
GST (nucleotide 969–1469) is a DNA fragment of
500
bp
(forward
primer:
5’-AAGTTCGCCCACTT
CACCGCAAATC-3’
and
reverse
primer:
5’-TCGAG
GAATGAGCCGGAGTTGTGGT-3’). The hpRNA binary
vectors
were
introduced
into
Agrobacterium
strain
EHA105 by electroporation. RNA analysis This protocol was also
used successfully for transient expression of the uidA
gene in the roots of rice plants grown under hydropo-
nics (data not shown). the QPCR reaction. First strand cDNA was synthesized
by reverse transcription using 1.5 μg of total RNA
in
20
μl
of
reaction volume using SuperScriptIII
(Invitrogen) as per manufacturer's instructions. Diluted
cDNA samples (1/10) were used for Real time PCR
analysis with 200 mM of each gene specific primer
mixed with SYBR Green PCR master mix in a final
volume of 15 μl following manufacturer's instructions. The reaction was carried out in 96-well optical reaction
plates (Roche), using the Light Cycler 480 Sequence
Detection System and software (Roche). Primer sequences were: 5’-pds, 5’-GTTCCTGATCGAGT
GAACGATG-3’, 3’-pds, 5’-CGAACATGGTCAACAATAGGC-
3’; 5’-OsExp, 5’-CGGTTAGCTAGAGTTCATGTGAGA-3’, and
3’-OsExp, 5’-ATTGGAGTAGTGGAGTGCCAAA-3’. Analysis of RNAi in HpRNA T-DNA stable transformants
To determine whether RNA silencing can be initiated in
agro-infected rice leaves by our method, we selected two
endogenous gene targets namely Phytoene Desaturase
(OsPDS) and SLENDER 1 (OsSLR1 or OsGAI, a rice
ortholog of the height-regulating gene GAI/RGA/RHT/
D8) because of their well-documented loss-of function
phenotype. PDS has been used extensively to investigate
gene silencing both in dicots (Voinnet 2001) and mono-
cots (Miki et al. 2005). We first analyzed the endogenous
mRNA level present in different tissues of wild type Nip-
ponbare plants. We carried out northern blot analyses
using the sequences of OsPDS and OsSLR1 Gene Spe-
cific Tags (GST) as probes to hybridize sequentially the
same membranes. In callus and leaves of six day-old
seedlings, transcript accumulation of OsPDS and OsSLR1
were rather similar. Contrastingly, in leaves of 5–6 leaf
stage plantlets, OsPDS mRNAs accumulated at a higher
level than those of OsSLR1 (Figure 3A). To determine
the efficacy of the hpRNA construct in triggering RNA
silencing and thereby conducting to the anticipated loss
of function phenotype, stable transgenic rice lines were
produced through co-culture of 25 embryogenic calli of
cv Nipponbare. Analysis of 85 OsPDS RNAi T0 lines
plants showed various levels of PDS mRNA degradation,
leading to phenotypes ranging from wild type to fully
albino plants, in which the PDS gene was found to be
strongly silenced (Figure 3B). Primary 21-nts-siRNAs, a
molecular marker for dsRNA-based gene silencing, were
easily detected in all albino lines which accounted for
54% of the regenerated plants (Figure 3C). RNA analysis Total RNA extraction was performed with Tri-Reagent
(Sigma) and was used for both high-molecular-weight
and small RNA analysis. Total RNA were separated
on
17.5%
denaturing
polyacrylamid
gels
and
then
transferred to nylon membranes (Hybond NX, Amer-
sham). Perfect-Hyb
buffer
(Sigma)
was
used
for
hybridization. Probes were DNA fragments labelled
by
random
priming
incorporation
of
[32
P]dCTP pBIOS738 binary vector
pC5300-UGN binary vector
Tnos
attR2
attR1
ccdB
Os Tubulin
intron
attR1
attR2
ccdB
CmR
pCsVMV
RB
LB
Sc4A
Os Actin
intron
nptII
TCaMV
Tnos
uidA
pUBI
RB
LB
pCaMV
Catalase
intron
hpt
TCaMV
A
B
Ubi1 intron
Figure 1 Schematic maps of the pBIOS738 (W. Paul, Biogemma) and pC5300-UGN T-DNAs. (A) The inverted repeat gateway cassette
cloned between the O. sativa tubulin intron is under the control of the strong constitutive promoter of the cassava vein mosaic virus (CsVMV). Sc4A: promoter of the fourth subunit of the subterranean clover mosaic virus. nptII: neomycin phospho transferase II: Tnos : terminator of the
Agrobacterium nopaline synthase gene: tCaMV : 3’ sequence of the Cauliflower Mosaic Virus (B) The core structure of the pC5300-UGN T-DNA. pUbi: promoter of the maize polyubiquitin 1 gene. uidA: E. coli β-glucuronidase coding sequence. A Os Tubulin
intron B pC5300-UGN binary vector Figure 1 Schematic maps of the pBIOS738 (W. Paul, Biogemma) and pC5300-UGN T-DNAs. (A) The inverted repeat gateway cassette
cloned between the O. sativa tubulin intron is under the control of the strong constitutive promoter of the cassava vein mosaic virus (CsVMV). Sc4A: promoter of the fourth subunit of the subterranean clover mosaic virus. nptII: neomycin phospho transferase II: Tnos : terminator of the
Agrobacterium nopaline synthase gene: tCaMV : 3’ sequence of the Cauliflower Mosaic Virus (B) The core structure of the pC5300-UGN T-DNA. pUbi: promoter of the maize polyubiquitin 1 gene. uidA: E. coli β-glucuronidase coding sequence. Page 4 of 12 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 two-fold concentration of acetosyringone (200 μM) also
increased GUS staining intensity. For all rice genotype
tested, high levels of GUS activity were observed 2 to
4 days following soaking in Agrobacterium cell suspen-
sions of EHA105, LBA4404 and AGL1 strains. GUS
staining was not detected any more after 10 days
(Figure 2C). No GUS staining was observed in un-
wounded
and
soaked
leaves
and
in
wounded
but
unsoaked leaves (not shown). RNA analysis A correlation
was observed between the accumulation of siRNA and
severity of the phenotype. Though a similar number of
rice embryogenic calli was co-cultivated with Agrobac-
terium bearing the OsSLR1 hpRNA construct, only four
plants regenerated from transformed cell lines displayed
a strong mutant phenotype but which sharply contrasted
with that of slr1 (Figure 3B) while other regenerated
lines displayed a normal WT phenotype. Mutant plants
were found to accumulate siRNAs whereas WT regene-
rants did not (Figure 3C). Based on the phenotype Agroinfection of rice leaf tissues with an A. tumefaciens
suspension (C) GUS staining observed after 1, 3, 6 and 10 days post agroinfection (d.p.a) for three different cultivars
of rice. No GUS staining was observed in unwounded and soaked leaves and in wounded but unsoaked leaves (not shown). A
B
12 cm
3 cm A B 1 d.p.a
cv. Nipponbare
3 d.p.a
6 d.p.a
10 d.p.a
C C cv. Zhong Zuo 321 cv. IR64 cv. Nipponbare Figure 2 Agroinfection of rice leaf tissues with A. tumefaciens suspension. Histochemical assays to assess the expression of the uidA gene in
rice tissues were carried out by staining with 5-bromo-4-chloro-3-indolyl-beta-D-glucuronid acid as described by Jefferson (1987). (A) Schematic
representation of the small apparatus carrying many needles used to wound rice leaves. (B) Wounded rice leaves soaking in the Agrobacterium
suspension in containment greenhouse. (C) GUS staining observed after 1, 3, 6 and 10 days post agroinfection (d.p.a) for three different cultivars
of rice. No GUS staining was observed in unwounded and soaked leaves and in wounded but unsoaked leaves (not shown). Figure 2 Agroinfection of rice leaf tissues with A. tumefaciens suspension. Histochemical assays to assess the expression of the uidA gene in
rice tissues were carried out by staining with 5-bromo-4-chloro-3-indolyl-beta-D-glucuronid acid as described by Jefferson (1987). (A) Schematic
representation of the small apparatus carrying many needles used to wound rice leaves. (B) Wounded rice leaves soaking in the Agrobacterium
suspension in containment greenhouse. (C) GUS staining observed after 1, 3, 6 and 10 days post agroinfection (d.p.a) for three different cultivars
of rice. No GUS staining was observed in unwounded and soaked leaves and in wounded but unsoaked leaves (not shown). transcription of the hpRNA construct, we agroinfected
leaves of wild type Nipponbare plants using the T-DNA
construct carrying the OsPDS and OsSLR1 hpRNA
structures. pBIOS738 empty vector was used as control
over the time course experiment. RNA was isolated
from the agro-infected regions of the leaf and analysed
for the accumulation of transcripts and of siRNAs, 2,
4, 8, 12 and 20 days following inoculation (Figure 4A
and B). Each time point involved at least three bio-
logical replicates. In the agro-infected region of the
leaf, OsPDS and OsSLR1 siRNAs were consistently
detected from day 2 post-agroinfection. The image
analysis software, ImageQuantTM TL (Amersham), was
used to quantify siRNAs detected by northern blot-
ting (Figure 4C). Agroinfection of rice leaf tissues with an A. tumefaciens
suspension To establish an efficient method for Agrobacterium in-
fection of rice leaves, we tested different A. tumefaciens
strains
(GV2200,
EHA105,
AGL1,
LBA4404,
and
GV3101) carrying the same binary plasmid as well as
different
infiltration
media
and
temperature,
and
assessed the efficiency of T-DNA transfer by expression
of the carried uidA gene by GUS histochemical assays. The infiltration media and method of Agrobacterium
preparation used for Arabidopsis thaliana floral dip or
Nicotiana leaf agroinfiltration were completely ineffi-
cient on rice leaves whatever temperature or bacterial
cell density applied (data not shown). Only the method
of bacteria preparation used for genetic transformation
of rice embryogenic calli yielded good results, using
hyper-virulent
strains
such
as
EHA105,
AGL1
or
LBA4404. Agrobacterium cells were plated on solid AB
medium containing antibiotic and incubated at 28°C for
3 days. The bacteria were then collected with a flat spa-
tula and diluted in liquid agroinfection medium to opti-
mal bacterial cell density ranging between 0.5 and 0.8. Because A. tumefaciens was not able to penetrate in the
leaf through epidermal cuticle or via sub-stomatal cham-
ber, we built a small apparatus carrying many 600 μm
diameter needles (Figure 2A) to quickly produce a large
number of wounds on the rice leaf surface. To avoid tis-
sue yellowing or wilting, rice leaves were incubated in
the Agrobacterium suspension for 30 to 60 minutes
(Figure 2B). The first experiments, carried out in the
containment greenhouse at 27-28°C, conducted to very
weak uidA transgene expression, upon GUS histoche-
mical
assay
(data
not
shown). Consequently,
the
temperature was reduced to 20°C during incubation. Moreover,
supplementation
of
the
agro-infection
medium with Silwet L-77, a powerful surfactant, proved
necessary to reach a high level, transgene expression. A Page 5 of 12 Andrieu et al. Rice 2012, 5:23 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 http://www.thericejournal.com/content/5/1/23 1 d.p.a
cv. Nipponbare
cv. IR64
cv. Zhong Zuo 321
3 d.p.a
6 d.p.a
10 d.p.a
C
A
B
12 cm
3 cm
Figure 2 Agroinfection of rice leaf tissues with A. tumefaciens suspension. Histochemical assays to assess the expression of the uidA gene in
rice tissues were carried out by staining with 5-bromo-4-chloro-3-indolyl-beta-D-glucuronid acid as described by Jefferson (1987). (A) Schematic
representation of the small apparatus carrying many needles used to wound rice leaves. (B) Wounded rice leaves soaking in the Agrobacterium
suspension in containment greenhouse. Agroinfection of rice leaf tissues with an A. tumefaciens
suspension The OsPDS siRNAs concentration
peaked at day 12 and dramatically decreased to reach
a very low level at day 20, in the two first experi-
ments. On the other hand, we were able to detect
OsPDS siRNA 30 days after agro-infection in a third
experiment (data not shown). Accumulation of the
OsSLR1 siRNAs was still occurring at day 20 though
likely reaching a plateau. In each experiment, a popula-
tion of plants agro-infected was further grown in the
containment greenhouse and phenotypic observations
were regularly performed. These plants displayed the description of slr1 mutants by Ikeda et al. (2001) we
were indeed expecting a constitutive gibberellin response
that leads to a tall phenotype. Though the OsSLR1 is
unique in the rice genome, the sequences of four other
gene members of the GRAS family (namely Os01g45860.1
also
called
SLRL1,
Os05g49990.1,
Os11g03110.1
and
Os12g06540.1) (Additional file 1: Figure S1) share high
similarity with a putative 21nt-siRNA residing in the
OsSLR1 GST sequence (Additional file 2: Figure S2-A). This makes them candidates prone to targeting by OsSLR1
primary siRNA that accumulate in the HpRNAi slr1
mutants (Additional file 2: Figure S2-B) through a me-
chanism called cross-silencing. Hybridization of small
RNA blots of the HpRNAi slr1 mutant with probes spe-
cific to each of these 4 genes evidenced accumulation of 21
nt siRNAs for 3 of them (Additional file 2: Figure S2-C). It
is therefore likely that this cross silencing accounts for
the contrasting phenotype observed in our HpRNAi slr1
mutant with regards to that of the reference disruption
mutant. Analysis of the RNA silencing in leaves following
agroinfection To determine whether agro-infection of leaf cells con-
ducts to local induction of RNA silencing following Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 Page 6 of 12 OsPDS
rRNA
Callus
6 day- old seedling
Leaf sheath
OsSLR1
rRNA
Callus
6 day- old seedling
Leaf sheath
L
L R
L R
Nb
phenotype
Albino
WT
C
B
OsPDS RNAi
mutants
SLR1 RNAi mutants
21 nt
OsPDS RNAi mutants
Nb
SLR1 RNAi mutants
mutant
WT
phenotype
RNA
24 nt
probe
GST
5’+3’
A
Figure 3 Analysis of HpRNA T-DNA stable transformants. (A) Northern blot detection of OsPDS and OsSLR1 mRNA accumulation in different
tissues of wild type plants of cv. Nipponbare. (B) Phenotypes of RNAi mutant lines expressing OsPDS RNAi HpRNA construct. (C) Detection of
21-nts siRNAs in OsPDS and OsSLR1 HpRNA transgenic lines exhibiting or not a mutant phenotype. A probe specific to the gene specific tag (GST)
and a probe representing regions extending 3’ and 5’ from the GST interval have been sequentially used. L (leaf), R (root), Nb (Control
Nipponbare plant). OsPDS
rRNA
Callus
6 day- old seedling
Leaf sheath
OsSLR1
rRNA
Callus
6 day- old seedling
Leaf sheath
B
OsPDS RNAi
mutants
SLR1 RNAi mutants
A A OsPDS
rRNA
Callus
6 day- o
Leaf she
OsSLR1
rRNA
Callus
6 day-
Leaf sh
B
OsPDS RNAi
mutants
SLR1 RNAi mutants B
OsPDS RNAi
mutants
SLR1 RNAi mutants B B
OsPDS RNAi
mutants
SLR L
L R
L R
Nb
phenotype
Albino
WT
C
21 nt
OsPDS RNAi mutants
Nb
SLR1 RNAi mutants
mutant
WT
phenotype
RNA
24 nt
probe
GST
5’+3’ SLR1 RNAi mutants
phenotype C Figure 3 Analysis of HpRNA T-DNA stable transformants. (A) Northern blot detection of OsPDS and OsSLR1 mRNA accumulation in different
tissues of wild type plants of cv. Nipponbare. (B) Phenotypes of RNAi mutant lines expressing OsPDS RNAi HpRNA construct. (C) Detection of
21-nts siRNAs in OsPDS and OsSLR1 HpRNA transgenic lines exhibiting or not a mutant phenotype. A probe specific to the gene specific tag (GST)
and a probe representing regions extending 3’ and 5’ from the GST interval have been sequentially used. L (leaf), R (root), Nb (Control
Nipponbare plant). japonica cv. Nipponbare. Throughout the experiment
and in both cultigens of rice, siRNA were not detected in
the non agro-infected leaves. same phenotype as the control plants throughout their
life cycle. Analysis of the RNA silencing in leaves following
agroinfection No siRNA could be detected in the tissues of
non agro-infected leaves of higher and lower rank, what-
ever the hpRNA construct used, suggesting that very low
or null long distance siRNA movement occurred through
the phloem. In order to test whether the pattern of
siRNA formation could be reproduced in indica rice, we
repeated the agro-infection experiment with the same
construct in leaves of cv. IR64. The siRNA accumulation
pattern, analysed over 30 days post agro-infection, was
fully consistent with that previously observed in the OsPDS and OsSRL1 silencing transitivity in rice leaf
tissues A parallel time course was achieved with leaf tissues of plants agro-infiltrated with the
pBIOS738 empty vector that served as reference value for siRNA accumulation. (C) Quantification of siRNAs detected by northern blotting using
the ImageQuant™TL image analysis software. genes. In both agro-infected leaf areas and in stably
transformed leaf tissues expressing the OsPDS hpRNA
construct, siRNAs corresponding to the region used to
produce the trigger dsRNA were detected but no siRNA
was detected when the regions extending either 5’ or 3’
from the trigger dsRNA borders were used as probe
(Figure 3C). The membranes were re-hybridized with
success using the trigger dsRNA probe to ascertain the
presence of siRNAs on the filters following repeated
cycles of hydridization-dehybridization. Contrastingly,
siRNA corresponding to regions extending 5’ and 3’
from the trigger RNA interval accumulated in stably
and transiently transformed leaf tissues expressing the
OsSLR1 hpRNA construct (Figures 3C and 5A and B). As the probes used were long the possibility of non-
specific cross reaction between the 5’ and 3’ probes and
the
siRNAs
derived
from
the
silencing
triggering
hpRNA could not be ruled out. In order to ascertain
hybridization specificity, the Ambion online siRNA tar-
get finder was used to identify all the potential siRNA
from 5’, GST and 3’ part of OsSLR1 coding sequence. Thirteen, 12, and 5 potential siRNAs were identified in
these 3 respective regions (Figure 5C). It is assumed
that nearly 50% of the siRNAs designed using this tool will achieve a >50% reduction of target gene expression. Considering this, oligo-nucleotides corresponding to
each group were mixed in equimolar ratio and used as
probe to re-hybridize small RNAs from leaves expres-
sing the OsSLR1 HpRNA construct (Figure 5D). Despite
detection of lower signals, these experiments confirmed
results obtained using longer probes. As a glimpse of
Ambion software efficiency, potential siRNAs detected
along the OsSLR1 GST sequence were sequentially used
as probe to re-hybridize RNAs extracted from slr1 mu-
tant lines. Five (41.6%) of 12 potential siRNAs showed a
clear hybridization signal (data not shown). OsPDS and OsSRL1 silencing transitivity in rice leaf
tissues In order to determine whether a transitive RNAi me-
chanism has been initiated in mutant lines and in agro-
infected rice leaves, we hybridized the available blots
with probes corresponding to regions the extending
5’ and 3’ from the trigger dsRNA interval in the two Page 7 of 12 Andrieu et al. Rice 2012, 5:23 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 http://www.thericejournal.com/content/5/1/23 B
2 4 8 12 20
0
0
5000
10000
15000
20000
25000
30000
35000
40000
0
2
4
8
12
20
d.p.a
Relative siRNA level
OsSLR1 siRNA
OsPDS siRNA
0
10000
20000
30000
40000
50000
60000
0
2
4
8
12
20
d.p.a
Relative siRNA level
21 nt
21 nt
A
Ef1
(495 bp)
OsSLR1 (318 bp)
OsPDS (383 bp)
C
Ef1
(495 bp)
2 4 8 12 20
0
2 4 8 12 20
0
2 4 8 12 20
0
d.p.a
d.p.a
d.p.a
d.p.a
Figure 4 Analysis of the agroinfiltrated leaf tissues. (A) Accumulation of OsPDS and OsSLR1 mRNAs revealed by RT-PCR during the time
course experiment. Ef1α mRNA was used as a standard. (B) Time course of OsPDS and OsSLR1 siRNAs accumulation following leaf agroinfection. Detection of siRNAs was performed using a GST probe. Arrow indicates the 21-nt-size-class siRNAs. Fifteen μg of total RNA extracted from leaf
tissues after 2, 4, 8, 12 and 20 days were loaded on gel. A parallel time course was achieved with leaf tissues of plants agro-infiltrated with the
pBIOS738 empty vector that served as reference value for siRNA accumulation. (C) Quantification of siRNAs detected by northern blotting using
the ImageQuant™TL image analysis software. A
Ef1
(495 bp)
OsSLR1 (318 bp)
OsPDS (383 bp)
Ef1
(495 bp)
2 4 8 12 20
0
2 4 8 12 20
0
d.p.a
d.p.a A B 0
5000
10000
15000
20000
25000
30000
35000
40000
0
2
4
8
12
20
d.p.a
Relative siRNA level
C
d.p.a C Figure 4 Analysis of the agroinfiltrated leaf tissues. (A) Accumulation of OsPDS and OsSLR1 mRNAs revealed by RT-PCR during the time
course experiment. Ef1α mRNA was used as a standard. (B) Time course of OsPDS and OsSLR1 siRNAs accumulation following leaf agroinfection. Detection of siRNAs was performed using a GST probe. Arrow indicates the 21-nt-size-class siRNAs. Fifteen μg of total RNA extracted from leaf
tissues after 2, 4, 8, 12 and 20 days were loaded on gel. Variation of OsPDS transcript accumulation along the
agroinfected leaf A
5’
3’
ATG
TAA
5’ probe
890 pb
GST probe
501 pb
3’ probe
368 pb
2 4 8 12 20
C
B
5’ probe
890 pb
GST probe
501 pb
3’ probe
368 pb
21 nt
2 4 8 12 20
C
2 4 8 12 20
C B C
AAGGTCGCTGCCTACTTCGGC
3
AAGCAGATCCCCACGCTGGCC
2
AACTTCGCGGCCGCGGAGGCG
1
AATGCGCCCGCCGTGCCGGTT
0
AAGGGGCCGCGGCGGCGAATG
9
AAGCGGATGCGCACTGGCGGC
8
AACTCCCAGCCGCTGCCGACT
7
AACGCGCCGCTGCCCCCTATC
6
AACCCCTCGGACCTCTCCTCC
5
AAGCTGGAGCAGCTGGAGATG
4
AAGGTGCGGTCGTCCGACATG
3
AAGGTGATGGCGGGGGCGGCG
2
AAGGACAAGGTGATGGCGGGG
1
5‘-SLR1
AAGGTCGCTGCCTACTTCGGC
3
AAGCAGATCCCCACGCTGGCC
2
AACTTCGCGGCCGCGGAGGCG
1
AATGCGCCCGCCGTGCCGGTT
0
AAGGGGCCGCGGCGGCGAATG
9
AAGCGGATGCGCACTGGCGGC
8
AACTCCCAGCCGCTGCCGACT
7
AACGCGCCGCTGCCCCCTATC
6
AACCCCTCGGACCTCTCCTCC
5
AAGCTGGAGCAGCTGGAGATG
4
AAGGTGCGGTCGTCCGACATG
3
AAGGTGATGGCGGGGGCGGCG
2
AAGGACAAGGTGATGGCGGGG
1
5‘-SLR1 C probe
10 30
C
C M
10 30
C
C M
10 30
C
C M
siRNA mix-5’
siRNA mix-GST
siRNA mix-3’
D
21 nt D Figure 5 Analysis of OsSLR1 silencing transitivity in agro infiltrated leaf tissues. (A) Schematic representation of the endogenous OsSLR1
gene with size and position of the different probes used to detect primary and secondary siRNAs. (B) Time-course analysis of primary (GST probe)
and secondary (5’ and 3’ probes) OsSLR1 siRNA after rice leave agroinfection. (C) Sequences of potential siRNAs detected with Ambion software
“siRNAs Target Finder” from 5’, GST and 3’ sequence part of OsSLR1 coding sequence. (D) Time-course analysis of primary (putative siRNAs mix
from GST as probe) and secondary (putative siRNAs mix from 5’ and 3’ part of OsSLR1 coding sequence as probes) OsSLR1 siRNA following rice
leaves agroinfection and in OsSLR1 RNAi mutant leaves (M). to the wild type, the quantity of OsPDS mRNA fell to 29%
two days after the agroinfection, and then reached a mini-
mum of 10% at day 4. mRNA accumulation in agro-
infected tissues reached 17% then 44% of the wild type
value at day 8 and 12 respectively (Figure 6B). Similar
results were observed for adjacent tissues with those
agroinfected with a 4 day-shift for the peak of degradation
of OsPDS mRNA which reaches 10% of the value of the
wild type 8 days following agro-infection. internal control gene to determine the relative abundance
of target gene transcripts in the leaf tissue. OsExp, similar
to its Arabidopsis ortholog (At4g33380), was chosen as the
internal control because this gene showed good stability
and its amplification remains in a log-linear stage at the
same optimal conditions as those of target genes. Variation of OsPDS transcript accumulation along the
agroinfected leaf To investigate the spreading of the RNAi silencing signal
along the rice leaf, we compared the mRNA level of the
target gene in agro-infected and adjacent leaf tissues by
quantitative
RT-PCR
(Figure
6A). As
the
HpRNA
construct-mediated silencing of the OsSLR1 gene was
prone to a complex cross silencing phenomenon that might
be specific to the gene, we focused this experiment on
tissues infiltrated with the OsPDS HpRNA construct. For
q-RT-PCR, the gene of interest is co-amplified with an Page 8 of 12 Andrieu et al. Rice 2012, 5:23 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 http://www.thericejournal.com/content/5/1/23 A
5’
3’
ATG
TAA
5’ probe
890 pb
GST probe
501 pb
3’ probe
368 pb
2 4 8 12 20
C
B
5’ probe
890 pb
GST probe
501 pb
3’ probe
368 pb
21 nt
2 4 8 12 20
C
2 4 8 12 20
C
probe
10 30
C
C M
10 30
C
C M
10 30
C
C M
siRNA mix-5’
siRNA mix-GST
siRNA mix-3’
D
21 nt
C
AAGGTCGCTGCCTACTTCGGC
3
AACTCCGGCTCATTCCTCGAC
AAGCAGATCCCCACGCTGGCC
2
AACCACAACTCCGGCTCATTC
AACTTCGCGGCCGCGGAGGCG
1
AAGGATCGTCACCGTGGTAGA
AATGCGCCCGCCGTGCCGGTT
0
AAGGTCCTGGGCACGGTGCAC
AAGGGGCCGCGGCGGCGAATG
9
AACTCGGTGTTCGAGCTGCAC
AAGCGGATGCGCACTGGCGGC
8
AACGAGGAGCCTGAGGTGATC
AACTCCCAGCCGCTGCCGACT
7
AAGCTTGCCCAGTTCGCGCAC
AACGCGCCGCTGCCCCCTATC
6
AAGGAGGGCTGCCTCACGCTG
AAGCCATCCTCGAGGCTTTCG
AACCCCTCGGACCTCTCCTCC
5
AAACAGGCGAGCACGCTCCTC
AAATCAAGCCATCCTCGAGGC
AAGCTGGAGCAGCTGGAGATG
4
AATGCCTACAAACAGGCGAGC
AATGGCCAGCTCTCCTCCAGG
AAGGTGCGGTCGTCCGACATG
3
AACCGCCTCGGCCGCGCCGGC
AAGCAGGGGATGCAATGGCCA
AAGGTGATGGCGGGGGCGGCG
2
AACGTCGTGGCGTGCGAGGGC
AAGTTCGCCCACTTCACCGC
AAGGACAAGGTGATGGCGGGG
1
3‘-SLR1
GST-SLR1
5‘-SLR1
AAGGTCGCTGCCTACTTCGGC
3
AACTCCGGCTCATTCCTCGAC
AAGCAGATCCCCACGCTGGCC
2
AACCACAACTCCGGCTCATTC
AACTTCGCGGCCGCGGAGGCG
1
AAGGATCGTCACCGTGGTAGA
AATGCGCCCGCCGTGCCGGTT
0
AAGGTCCTGGGCACGGTGCAC
AAGGGGCCGCGGCGGCGAATG
9
AACTCGGTGTTCGAGCTGCAC
AAGCGGATGCGCACTGGCGGC
8
AACGAGGAGCCTGAGGTGATC
AACTCCCAGCCGCTGCCGACT
7
AAGCTTGCCCAGTTCGCGCAC
AACGCGCCGCTGCCCCCTATC
6
AAGGAGGGCTGCCTCACGCTG
AAGCCATCCTCGAGGCTTTCG
AACCCCTCGGACCTCTCCTCC
5
AAACAGGCGAGCACGCTCCTC
AAATCAAGCCATCCTCGAGGC
AAGCTGGAGCAGCTGGAGATG
4
AATGCCTACAAACAGGCGAGC
AATGGCCAGCTCTCCTCCAGG
AAGGTGCGGTCGTCCGACATG
3
AACCGCCTCGGCCGCGCCGGC
AAGCAGGGGATGCAATGGCCA
AAGGTGATGGCGGGGGCGGCG
2
AACGTCGTGGCGTGCGAGGGC
AAGTTCGCCCACTTCACCGC
AAGGACAAGGTGATGGCGGGG
1
3‘-SLR1
GST-SLR1
5‘-SLR1
Figure 5 Analysis of OsSLR1 silencing transitivity in agro infiltrated leaf tissues. (A) Schematic representation of the endogenous OsSLR1
gene with size and position of the different probes used to detect primary and secondary siRNAs. (B) Time-course analysis of primary (GST probe)
and secondary (5’ and 3’ probes) OsSLR1 siRNA after rice leave agroinfection. (C) Sequences of potential siRNAs detected with Ambion software
“siRNAs Target Finder” from 5’, GST and 3’ sequence part of OsSLR1 coding sequence. (D) Time-course analysis of primary (putative siRNAs mix
from GST as probe) and secondary (putative siRNAs mix from 5’ and 3’ part of OsSLR1 coding sequence as probes) OsSLR1 siRNA following rice
leaves agroinfection and in OsSLR1 RNAi mutant leaves (M). Discussion powerful surfactant. After testing different A. tumefaciens
strains as well as different infiltration media and tempera-
tures, assessed by GUS histochemical assays, we have deve-
loped an efficient and reproducible method for agro-
infection of leaves in indica and japonica rice, based on
soaking wounded tissues in EHA105 bacterial solution at
low temperature. The most critical physiological parameter
appeared
to
be
the
post
agro-infiltration
incubation
temperature. The largest and most intensively stained GUS
areas in leaves were observed when rice plants are trans-
ferred at 20°C following agro-infiltration. A hypothesis is
that a 25°C/28°C temperature favors bacterial multiplica-
tion vs. their attachment to plant cells and T-DNA
transfer whereas at 20°C, bacterial multiplication slows
down, thereby favoring plant cell-bacteria interactions. As
reported in the literature in other plants (Batra and Variation of OsPDS transcript accumulation along the
agroinfected leaf Primer sequences were: 5’-pds,
5’-GTTCCTGATCGAGTGAACGATG-3’, 3’-pds, 5’-CGAACATGGTCAACAATAGGC-3’; 5’-OsExp, 5’-CGGTTAGCTAGAGTTCATGTGAGA-3’, and 3’-OsExp,
5’-ATTGGAGTAGTGGAGTGCCAAA-3’. NS, *, **,and *** means non significantly different and statistically different at the 5%, 1% and 0.1% level in a
t-student test respectively. Figure 6 Analysis of the movement of the OsPDS silencing signal from agroinfiltrated leaf tissues. (A) Schematic representation of the
rice leaf. Wounded and agroinfected part of the leaf and adjacent part are indicated. (B) Real-time PCR time course analysis of OsPDS transcript
accumulation in the agro-infiltrated and adjacent region of the leaf. Results of 3 biological replicates are shown. Primer sequences were: 5’-pds,
5’-GTTCCTGATCGAGTGAACGATG-3’, 3’-pds, 5’-CGAACATGGTCAACAATAGGC-3’; 5’-OsExp, 5’-CGGTTAGCTAGAGTTCATGTGAGA-3’, and 3’-OsExp,
5’-ATTGGAGTAGTGGAGTGCCAAA-3’. NS, *, **,and *** means non significantly different and statistically different at the 5%, 1% and 0.1% level in a
t-student test respectively. Variation of OsPDS transcript accumulation along the
agroinfected leaf Analysis
of leaves agroinfected with the OsPDS hpRNA construct
showed a close relationship between hpRNA transient ex-
pression and depression of PDS transcripts level. Compared Andrieu et al. Rice 2012, 5:23 Page 9 of 12 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 http://www.thericejournal.com/content/5/1/23 0
0,2
0,4
0,6
0,8
1
1,2
C
2
4
8
12
Agroinfected tissue area
Adjacent tissue area
Days post agro infection
Relative mRNA levels
*
NS
*
**
***
Agroinfected part
of the leaf ( 10 cm)
Discarded
Adjacent part
of the leaf ( 2 cm)
A
B
Figure 6 Analysis of the movement of the OsPDS silencing signal from agroinfiltrated leaf tissues. (A) Schematic representation of the
rice leaf. Wounded and agroinfected part of the leaf and adjacent part are indicated. (B) Real-time PCR time course analysis of OsPDS transcript
accumulation in the agro-infiltrated and adjacent region of the leaf. Results of 3 biological replicates are shown. Primer sequences were: 5’-pds,
5’-GTTCCTGATCGAGTGAACGATG-3’, 3’-pds, 5’-CGAACATGGTCAACAATAGGC-3’; 5’-OsExp, 5’-CGGTTAGCTAGAGTTCATGTGAGA-3’, and 3’-OsExp,
5’-ATTGGAGTAGTGGAGTGCCAAA-3’. NS, *, **,and *** means non significantly different and statistically different at the 5%, 1% and 0.1% level in a
t-student test respectively. 0
0,2
0,4
0,6
0,8
1
1,2
C
2
4
8
12
Agroinfected tissue area
Adjacent tissue area
Days post agro infection
Relative mRNA levels
*
NS
*
**
***
Agroinfected part
of the leaf ( 10 cm)
Discarded
Adjacent part
of the leaf ( 2 cm)
A
B
Figure 6 Analysis of the movement of the OsPDS silencing signal from agroinfiltrated leaf tissues. (A) Schematic representation of the
rice leaf Wounded and agroinfected part of the leaf and adjacent part are indicated (B) Real time PCR time course analysis of OsPDS transcript Agroinfected part
of the leaf ( 10 cm)
Discarded
Adjacent part
of the leaf ( 2 cm)
A A 0
0,2
0,4
0,6
0,8
1
1,2
C
2
4
8
12
Agroinfected tissue area
Adjacent tissue area
Days post agro infection
Relative mRNA levels
*
NS
*
**
***
B B Days post agro infection Figure 6 Analysis of the movement of the OsPDS silencing signal from agroinfiltrated leaf tissues. (A) Schematic representation of the
rice leaf. Wounded and agroinfected part of the leaf and adjacent part are indicated. (B) Real-time PCR time course analysis of OsPDS transcript
accumulation in the agro-infiltrated and adjacent region of the leaf. Results of 3 biological replicates are shown. An Agrobacterium-mediated transient expression system
for cereal leaves Because they provide a rapid, versatile and convenient way
for achieving a very high level of gene expression in a dis-
tinct and defined zone of leaf, Agrobacterium-mediated
transient expression systems have been chosen for inducing
RNA silencing processes and transitivity analysis in leaves
(Schöb et al. 1997). The first objective of this study was to
evaluate the feasibility of an agro-infiltration approach for
transient expression of a T-DNA construct in leaf tissues of
rice. In cereal species, the leaf structure is known to prevent
infiltration of a bacterial solution by simple pressure. In
order to provide an access of Agrobacterium to the leaf
cells, we used a combination of multiple uniform wound
treatment and supplementation of the suspension with a Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 Page 10 of 12 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 expression of the T-DNA-borne genes occurs only during
the very few days following agroinfection (as attested by
the revelation by histochemical assay of the otherwise ra-
ther stable GUS protein, that dramatically decreased be-
tween 6 to 10 days after agroinfection (data not shown)),
we concluded that RNA silencing was maintained in rice
cells in the absence of the trigger hpRNA. Local initiation
of silencing produced preferentially primary 21-nt siRNA
and maintenance of siRNAs production during cell life
indicated that target genes are actively transcribed. This
result is consistent with previously published data. 21-nt
siRNAs are believed to guide mRNA cleavage, whereas 24-
nt siRNAs are believed to exclusively mediate chromatin
modification and transcriptional silencing by acting in a
RISC-like complex (Brodersen and Voinnet 2006). Kumar 2003; Wydro et al. 2006), we showed that aceto-
syringone was essential for efficient transfection, and the
use of a two-fold concentration compared to our rice
stable transformation procedure increases the efficiency
of in planta agroinfection. As reported in other plants
species (Wroblewski et al. 2005; Chabaud et al. 2003), the
choice of the Agrobacterium strain appeared critical be-
cause of specific host-bacteria interactions (Tzfira and
Citovsky 2002). In Agrobacterium–mediated in planta
transformation, the genotype is generally found to influence
T-DNA transfer and transgene expression. This
was
observed
for
instance
in
Arabidopsis
and
grapevine
(Wroblewski et al. 2005; Zottini et al. 2008; Santos-Rosa
et al. 2008). An Agrobacterium-mediated transient expression system
for cereal leaves However, in our study, all the rice genotypes
tested were found to be amenable to Agrobacterium-
mediated transient expression of the uidA gene in infil-
trated leaf cells. The optimized rice agroinfection proto-
col was also applied with success to sugarcane suggesting
that other graminaceous species are amenable to this
procedure (unpublished results). This method could be
also very useful for transgenic complementation (Van der
Hoorn et al. 2000; Shao et al. 2003; Bertazzon et al. 2012),
promoter analysis (Yang et al. 2000), protein production
(Vaquero et al. 1999) and ascertaining miRNA and
amiRNA processing and target cleavage (Warthmann
et al. 2008) in rice plants. The method is also applicable at
different developmental stages as suggested by similar in-
tensities of GUS staining observed in leaves of plants from
the 5 leaf stage to the flowering stage. Our results indeed
pointed that the developmental stage of the rice plants is
not a critical parameter for an efficient expression of uidA
gene induced in agroinfected leaves, though the efficiency
becomes lower in old leaves than in seedling leaves prob-
ably due to plant resistance to agro-infection acquired at
maturity (Wroblewski et al. 2005; Wydro et al. 2006;
Santos-Rosa et al. 2008). Real-time PCR analysis showed the highest OsPDS tran-
script depletion in the agro-infected leaf area 4 days follow-
ing agro-infiltration. These results are consistent with the
literature which reports a transitory peak of transgene
expression at day 4 following agro-infection in N. Benthamiana and A. thaliana. Decrease of OsPDS
mRNA degradation parallels the decline of transient ex-
pression of the HpRNA construct. Longer term mainten-
ance of the silencing of the OsPDS gene and exhibition of
the
photobleaching
phenotype
would
have
probably
required higher siRNA accumulation in agro infiltrated
tissues. High level siRNA could be obtained in enhancing
Agrobacterium-mediated delivery in leaf cells or through
an amplification of the production of siRNA resulting from
transitivity mechanism (see following section). Using the
same hpRNA construct bearing the OsPDS GST sequence
to initiate RNAi in tobacco leaves following agro-infection,
we observed a similar amplification of the siRNAs and
their maintenance in tissues where the initiation took
place, but resulting in a photobleaching phenotype (unpub-
lished results). The most obvious difference between rice
and tobacco, at the experimental level, resides in the higher
penetration of agrobacteria in leaves and consequently of
bacterial cells in contact with leaf cells. An Agrobacterium-mediated transient expression system
for cereal leaves In order to obtain
an amplification of the degradation of the mRNA and to
observe a phenotype in rice leaves, it might be necessary to
increase the level of agro-infection in rice tissues. HpRNA constructs triggers a maintained RNAi process in
agro-infiltrated leaf tissues In the experiments described here, we have analysed
RNA silencing of two endogenous rice genes in both
agro-infected leaf tissues and stably transformed rice
plants, constitutively expressing an hpRNA construct. In
both situations, we were able to detect siRNAs corre-
sponding to the trigger dsRNA. In stable transgenic lines,
the accumulation of which correlated the penetrance of
the RNAi mutant phenotype, demonstrating the efficiency
of our hpRNA construct in initiating RNA silencing of the
corresponding endogenous gene in rice cells. Transient ex-
pression of hpRNA after leaf agro-infection was sufficient
to trigger RNA silencing. During 10 to 20 days following
rice agro-infection depending on the gene targeted, the
quantity of siRNAs increased in cells then remains stable
or decreased till leaf senescence. Considering that transient Conclusion In conclusion, this work demonstrates that in planta,
Agrobacterium-mediated transient expression of T-DNA
constructs is achievable in rice leaves. The possibility to
trigger RNAi in a localized manner in the rice leaf opens
new perspectives to study the cell-to-cell movement of
the silencing signal in a monocot plant. This method
could also be of particular interest to test the functional-
ity of natural and artificial miRNA expression construct,
in ascertaining miRNA processing and cleavage of target
genes that are expressed in rice leaves. It is indeed often
necessary to assay several amiRNAs, the sequences of
which are based on computational predictions, to find
one which effectively triggers knock down of a target
gene (Warthmann et al. 2008). As this transfection
method has been used to achieve localized GUS expres-
sion in roots of rice plants grown in hydroponics (data
not shown) it should be as well applicable for target
genes expressed in other rice organs. Spreading of the silencing signal to the non-agro
infiltrated leaf tissues Though the current model of cell-to-cell movement of
the RNA silencing signal implies a central role for tran-
sitivity (Himber et al. 2003), we surprisingly detected an
efficient decrease of the OsPDS mRNA level in the leaf
tissue adjacent to the agroinfiltrated area occuring with
a time lag. A possible explanation lies in the transfer of
the siRNA from agro-infected tissues to healthy tissues
through the symplastic pathway as described by Dunoyer
et al. (2010). The vast majority of experimental data
used to formulate the current model of cell-to-cell
movement of RNA silencing signal was accumulated in
N. benthamiana and A. thaliana. The short range move-
ment (up to 10–15 cells) of the silencing signal requires
accumulation of 21nt but not of 24nt siRNA while
the possibility that ssRNA move from one cell to the
other cannot be yet ruled out. The longer range (beyond
10–15
cells)
systemic
signal
appears
to
be
RDR6
dependent since no spreading and no secondary siRNA
is observed in rdr6 background. However, that long
range spreading has so far been only observed in the
RNAi of exogenous transgenes that generally implies ex-
tensive production of secondary siRNA and may result
in
exogenous
DNA
methylation. SiRNA cell-to-cell
movements occur either in a non selective or selective Aurélie Andrieu and Jean Christophe Breitler are first
co authors. HpRNA-mediated induction of OsPDS and OsSLR1
silencing differs by the occurrence of transitivity in both
stably and transiently transformed leaf tissues However, northern blot analysis of total RNA
extracted from wild type plants clearly showed that
transcription level of OsPDS is rather equivalent to
that of OsSLR1 in seedling leaves and is even higher at
a later developmental stage. Along the same line, silen-
cing of the most abundant mRNA in plants, that of
the rubisco small subunit, does not involves transitivity
nor long range spreading of the silencing signal (Vaistij
et al. 2002; Himber et al. 2003) suggesting that the
mRNA level might not be an essential factor in transi-
tive RNA silencing. Experience on the RNA silencing
transitivity in monocotyledonous plants is limited to
the pioneering report of Miki et al. (2005) whose
showed that the transitive RNA silencing machinery is
conserved in rice. In this study, whereas transitive
RNA silencing occurred in both 5’ and 3’ orientations
for
the
exogenous
gfp
gene,
no
transitivity
was
observed
for
the
endogenous
OsPDS
and
OsRAC
genes. Therefore, the cause of transitivity of the RNAi
signal in OsSLR1 remains to be further investigated. manner through plasmodesmata (Hyun et al. 2011). It is
thought that a molecular link exists between RNAi-
mediated DNA methylation and regulation of selective
siRNA movement through plasmodesmata but that link
remains to be elucidated. Cross silencing occurred in HpRNA silencing of OsSLR1
As demonstrated by Miki et al. (2005) with the OsRAC
gene family, a single IR of a specific region shared by se-
veral genes of a same family can simultaneously suppress
the expression of multiple genes. In this work we studied
the possibility of cross extinction of rice genes sharing se-
quence homology outside the sequence of OsSLR1 GST. The results showed that transitivity could be responsible
of multiple endogenous genes knock-down. So the choice
of a highly conserved and specific sequence, is necessary
to precisely suppress only one member in a gene family. HpRNA-mediated induction of OsPDS and OsSLR1
silencing differs by the occurrence of transitivity in both
stably and transiently transformed leaf tissues We were not able to evidence transitivity in both tran-
siently and stably silenced leaf tissues expressing the
OsPDS HpRNA construct. This result is consistent with
previous reports in tobacco (rubisco and pds genes;
Vaistij et al. 2002), Arabidopsis (rubisco and sulfur
genes; Himber et al. 2003) and rice (OsGen-L, Moritoh
et al. 2005; Ospds and OsRAC genes, Miki et al. 2005). Surprisingly, HpRNA-mediated silencing of the OsSLR1 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 Page 11 of 12 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 gene clearly involved transitivity with formation of
siRNA occurring from regions of the endogenous tar-
get mRNA extending 5’ and 3’ from the region con-
taining the trigger siRNA. One possible explanation
for apparent lack of transitive RNA silencing of OsPDS
endogenous gene may lie in a low concentration of
the
RNA
substrates
for
RDR
(Garcia-Perez
et
al. 2004). However, northern blot analysis of total RNA
extracted from wild type plants clearly showed that
transcription level of OsPDS is rather equivalent to
that of OsSLR1 in seedling leaves and is even higher at
a later developmental stage. Along the same line, silen-
cing of the most abundant mRNA in plants, that of
the rubisco small subunit, does not involves transitivity
nor long range spreading of the silencing signal (Vaistij
et al. 2002; Himber et al. 2003) suggesting that the
mRNA level might not be an essential factor in transi-
tive RNA silencing. Experience on the RNA silencing
transitivity in monocotyledonous plants is limited to
the pioneering report of Miki et al. (2005) whose
showed that the transitive RNA silencing machinery is
conserved in rice. In this study, whereas transitive
RNA silencing occurred in both 5’ and 3’ orientations
for
the
exogenous
gfp
gene,
no
transitivity
was
observed
for
the
endogenous
OsPDS
and
OsRAC
genes. Therefore, the cause of transitivity of the RNAi
signal in OsSLR1 remains to be further investigated. gene clearly involved transitivity with formation of
siRNA occurring from regions of the endogenous tar-
get mRNA extending 5’ and 3’ from the region con-
taining the trigger siRNA. One possible explanation
for apparent lack of transitive RNA silencing of OsPDS
endogenous gene may lie in a low concentration of
the
RNA
substrates
for
RDR
(Garcia-Perez
et
al. 2004). Additional file 1: Figure S1. Phylogenetic tree of the members of the
Arabidopsis thaliana and Oryza sativa GRAS transcription factor families
generated through the GreenphylDB GOST tool (http://greenphyl.cirad.fr/
v2/cgi-bin/index.cgi). Position of the four GRAS genes exhibiting a shared
putative siRNA with OsSLR1 are highlighted. Author details
1 1CIRAD, UMR AGAP, TAA108/03, Av Agropolis, F-34398 Montpellier Cedex 05,
France. 2Université Montpellier II, UMR DIADE, F-34398 Montpellier Cedex 05,
France. 1CIRAD, UMR AGAP, TAA108/03, Av Agropolis, F-34398 Montpellier Cedex 05,
France. 2Université Montpellier II, UMR DIADE, F-34398 Montpellier Cedex 05,
France. Schweizer P, Pokorny J, Schulze-Lefert P, Dudler R (2000) Double-stranded RNA
interferes with gene function at the single-cell level in cereals. Plant J 24:895–903 Sijen T, Fleenor J, Simmer F, Thijssen K, Parrish S, Timmons L, Plasterk R, Fire A
(2001) On the role of RNA amplification in dsRNA-triggered gene silencing. Cell 107:465–476 Received: 25 November 2011 Accepted: 12 July 2012
Published: 31 August 2012 References Tzfira T, Citovsky V (2002) Partners-in-infection: host proteins involved in the
transformation of plant cells by Agrobacterium. Cell Biol 12:121–129 Batra S, Kumar S (2003) Agrobacterium-mediated transient GUS gene expression
in buffel grass (Cenchrus ciliaris L.). J Appl Genet 44:449–458 Batra S, Kumar S (2003) Agrobacterium-mediated transient GUS gene expression
in buffel grass (Cenchrus ciliaris L.). J Appl Genet 44:449–458 Vaistij FE, Jones L, Baulcombe DC (2002) Spreading of RNA targeting and
DNA methylation in RNA silencing requires transcription of the target
gene and a putative RNA-dependent RNA polymerase. Plant Cell 14:857–867 Bertazzon N, Raiola A, Castiglioni C, Gardiman M, Angelini E, Borgo M, Ferrari S Bertazzon N, Raiola A, Castiglioni C, Gardiman M, Angelini E, Borgo M, Ferrari S
(2012) Transient silencing of the grapevine gene VvPGIP1 by agroinfiltration
with a construct for RNA interference. Plant Cell Rep 31:133–143 Bertazzon N, Raiola A, Castiglioni C, Gardiman M, Angelini E, Borgo M, Ferrari S
(2012) Transient silencing of the grapevine gene VvPGIP1 by agroinfiltration
with a construct for RNA interference. Plant Cell Rep 31:133–143 (2012) Transient silencing of the grapevine gene VvPGIP1 by agroinfiltration
with a construct for RNA interference. Plant Cell Rep 31:133–143 Vaquero C, Sack M, Chandler J, Drossard J, Schuster F, Monecke M, Schillberg S,
Fischer R (1999) Transient expression of a tumor-specific single-chain
fragment and a chimeric antibody in tobacco leaves. Proc Natl Acad Sci U S A
96:11128–11133 p
Breitler JC, Meynard D, Van Boxtel J, Royer M, Bonnot F, Cambillau L, Guiderdoni Breitler JC, Meynard D, Van Boxtel J, Royer M, Bonnot F, Cambillau L, Guiderdoni
E (2004) A novel 2 T-DNA binary vector allows efficient generation of marker-
free transgenic plants in three elite cultivars of rice (Oryza sativa L.). Transgenic Res 13:271–287 Van der Hoorn RA, Laurent F, Roth R, De Wit PJ (2000) Agroinfiltration is a
versatile tool that facilitates comparative analyses of Avr9/Cf-9-induced
and Avr4/Cf-4-induced necrosis. Mol Plant-Microbe Interact 13:439–446 Brodersen P, Voinnet O (2006) The diversity of RNA silencing pathways in plants. Trends Genet 22:268–280 Brodersen P, Voinnet O (2006) The diversity of RNA silencing pathways in plants. Trends Genet 22:268–280 Caldana C, Scheible W-R, Mueller-Roeber B, Ruzicic S (2007) A quantitative RT-PCR
platform for high-throughput expression profiling of 2,500 rice transcription
factors. Plant Methods 3:7–16 Verdaguer B, de Kochko A, Fux CI, Beachy RN, Fauquet C (1998) Functional
organization of the cassava vein mosaic virus (CsVMV) promoter. Competing interests
The authors declare they have no competing interest. Moritoh S, Miki D, Akiyama M, Kawahara M, Izawa T, Maki H, Shimamoto K
(2005) RNAi-mediated silencing of OsGEN-L (OsGEN-like), a new member of
the RAD2/XPG nuclease family, causes male sterility by defect of microspore
development in rice. Plant Cell Physiol 46:699–715 References Plant Mol
Biol 37:1055–1067 Chabaud M, De Carvalho-Niebel F, Barker DG (2003) Efficient transformation of
Medicago truncatula cv. Jemlong using the hypervirulent Agrobacterium
tumefaciens strain AGL1. Plant Cell Rep 22:46–51 Voinnet O (2001) RNA silencing as a plant immune system against viruses. Trends in Genetics 17:449–459 Voinnet O (2001) RNA silencing as a plant immune system against viruses. Trends in Genetics 17:449–459 Voinnet O (2001) RNA silencing as a plant immune system against viruses. Trends in Genetics 17:449–459
Voinnet O (2005a) Non-cell autonomous RNA silencing. FEBS Letters 579:5858–5871
Voinnet O (2005b) Induction and suppression of RNA silencing: insights from
viral infections. Nat Rev Genet 6:206–220
Warthmann N, Chen H, Ossowski S, Weigel D, Hervé P (2008) Highly Specific
Gene Silencing by Artificial miRNAs in Rice. PLoS One 3:e1829
Wroblewski T, Tomczak A, Michelmore R (2005) Optimization of Agrobacterium
-mediated transient assays of gene expression in lettuce, tomato and
Arabidopsis. Plant Biotech J 3:259–273
Wydro M, Kozubek E, Lehmann P (2006) Optimization of transient Agrobacterium Voinnet O (2005a) Non-cell autonomous RNA silencing. FEBS Letters 579:5858–5871
Voinnet O (2005b) Induction and suppression of RNA silencing: insights from
viral infections. Nat Rev Genet 6:206–220 Voinnet O (2005a) Non-cell autonomous RNA silencing. FEBS Letters 579:5858–5871
Voinnet O (2005b) Induction and suppression of RNA silencing: insights from
viral infections. Nat Rev Genet 6:206–220 Chilton M, Currier T, Farrand S, Bendich A, Gordon M, Nester E (1974)
Agrobacterium tumefaciens DNA and PS8 bacteriophage DNA not detected
in crown gall tumors. Proc Nat Acad Sci USA 71:3672–3676 Warthmann N, Chen H, Ossowski S, Weigel D, Hervé P (2008) Highly Specific
Gene Silencing by Artificial miRNAs in Rice. PLoS One 3:e1829 Christensen A, Quail P (1996) Ubiquitin promoter-based vectors for high level
expression of selectable and/or screenable marker genes in
monocotyledonous plants. Transgenic Res 5:213–218 Wroblewski T, Tomczak A, Michelmore R (2005) Optimization of Agrobacterium
-mediated transient assays of gene expression in lettuce, tomato and
Arabidopsis. Plant Biotech J 3:259–273 Dunoyer P, Schott G, Himber C, Meyer D, Takeda A, Carrington JC, Voinnet O
(2010) Small RNA duplexes function as mobile silencing signals between
plant cells. Science 328:912 Wydro M, Kozubek E, Lehmann P (2006) Optimization of transient Agrobacterium-
mediated gene expression system in leaves of Nicotiana benthamiana. Acta
Biochimica Polonica 53:289–298 Wydro M, Kozubek E, Lehmann P (2006) Optimization of transient Agrobacterium-
mediated gene expression system in leaves of Nicotiana benthamiana. Authors’ contributions AA carried out the work of agroinflitration and molecular analyses and
co-wrote the manuscript. JCB initiated the research, carried out the work of
agroinflitration and molecular analyses with AA and co-wrote the
manuscript. CS assisted JCB in siRNA blotting. DM produced stable
transformants of OsPDS and OsSLR1. PG supervised the work and edited the
manuscript. EG supervised the work and co-wrote the manuscript. All
authors read and approved the final manuscript. Nishikura K (2001) A short primer on RNAi: RNA-directed RNA polymerase acts as
a key catalyst. Cell 107:415–418 Sallaud C, Meynard D, van Boxtel J, Gay C, Bes M, Brizard JP, Larmande P,
Ortega D, Raynal M, Portefaix M, Ouwerkerk PB, Rueb S, Delseny M,
Guiderdoni E (2003) Highly efficient production and characterization of
T-DNA plants for rice (Oryza sativa L.) functional genomics. Theor Appl
Genet 106:1396–1408 Received: 25 November 2011 Accepted: 12 July 2012
Published: 31 August 2012 Small I (2007) RNAi for revealing and engineering plant gene functions. Cur Opin
Biotech 18:148–153 Competing interests Miki D, Itoh R, Shimamoto K (2005) RNA silencing of single and multiple
members in a gene family of rice. Plant Physiol 138:1903–1913 Miki D, Itoh R, Shimamoto K (2005) RNA silencing of single and multiple
members in a gene family of rice. Plant Physiol 138:1903–1913 Acknowledgments Santos-Rosa M, Poutaraud A, Merdinoglu D, Mestre P (2008) Development of a
transient expression system in grapevine via agro-infiltration. Plant Cell Rep
27:1053–1063 We thank the Région Languedoc Roussillon for providing AA with a PhD
grant. We thank Dr W. Paul and Dr P. Perez for the pBIOS 738 plasmid and
AGRIDYNE S.A. company for providing us with Silwet L-77 surfactant. We
thank Dr C. Périn, CIRAD, for valuable advices in the course of this research
and assistance in Q-RT-PCR analyses. Shao F, Golstein C, Ade J, Stoutemyer M, Dixon JE, Innes RW (2003) Cleavage of
Arabidopsis PBS1 by a bacterial type III effector. Science 301:1230–1233 Schöb H, Kunz C, Meins F (1997) Silencing of transgenes introduced into leaves
by agroinfiltration: a simple, rapid method for investigating sequence
requirements for gene silencing. Mol Gen Genet 256:581–585 Additional files Additional file 1: Figure S1. Phylogenetic tree of the members of the
Arabidopsis thaliana and Oryza sativa GRAS transcription factor families
generated through the GreenphylDB GOST tool (http://greenphyl.cirad.fr/
v2/cgi-bin/index.cgi). Position of the four GRAS genes exhibiting a shared
putative siRNA with OsSLR1 are highlighted. Additional file 2: Figure S2. (A) Alignments of the shared putative
21nt-siRNA residing in the sequence of OsSLR1and in those of the four
other members of the GRAS family. (B) Detection of the common
putative 21nt-siRNA in transformation events harbouring the OsSLR1
HpRNA T-DNA construct and exhibiting (mutant) or not (wt) a
phenotype. Nb: a control Nipponbare plant. (C) Detection of siRNAs
specific to four other members of the GRAS family the sequence of
which contains the putative, shared 21nt-siRNA in transformation events Page 12 of 12 Page 12 of 12 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 Andrieu et al. Rice 2012, 5:23
http://www.thericejournal.com/content/5/1/23 ortholog of the height-regulating gene GAI/RGA/RHT/D8. Plant Cell
13(5):999–1010 harbouring the OsSLR1 HpRNA T-DNA construct and exhibiting (mutant)
or not (wt) a phenotype. Nb: Control Nipponbare plant. Jefferson R (1987) Assaying chimeric genes in plants: The GUS gene fusion
system. Plant Mol Biol Rep 5:387–405 References Acta
Biochimica Polonica 53:289–298 Garcia-Perez R, Van Houdt H, Depicker A (2004) Spreading of post-transcriptional
gene silencing along the target gene promotes systemic silencing. Plant J
38:594–602 Yang Y, Li R, Qi M (2000) In vivo analysis of plant promoters and transcription
factors by agroinfiltration of tobacco leaves. Plant J 22:543–551 Ghildiyal M, Zamore PD (2009) Small silencing RNAs an expanding universe. Nat
Rev Genet 10(2):94–108 Zottini M, Barizza E, Costa A, Formentin E, Ruberti C, Carini F, Lo Schiavo F (2008)
Agroinfiltration of grapevine leaves for fast transient assay of gene
expression and for long-term production of stable transformed cells. Plant
cell rep 27:845–854 Himber C, Dunoyer P, Moissiard G, Ritzenthaler C, Voinnet O (2003) Transitivity-
dependent and -independent cell-to-cell movement of RNA silencing. EMBO
J 22:4523–4533 Hyun KT, Uddin MN, Rim Y, Kim JY (2011) Cell-to-cell trafficking of RNA and RNA
silencing through plasmodesmata. Protoplasma 248:101–116 doi:10.1186/1939-8433-5-23
Cite this article as: Andrieu et al.: An in planta, Agrobacterium-mediated
transient gene expression method for inducing gene silencing in rice
(Oryza sativa L.) leaves. Rice 2012 5:23. doi:10.1186/1939-8433-5-23
Cite this article as: Andrieu et al.: An in planta, Agrobacterium-mediated
transient gene expression method for inducing gene silencing in rice
(Oryza sativa L.) leaves. Rice 2012 5:23. Ikeda A, Ueguchi-Tanaka M, Sonoda Y, Kitano H, Koshioka M, Futsuhara Y,
Matsuoka M, Yamaguchi J (2001) Slender rice, a constitutive gibberellin
response mutant, is caused by a null mutation of the SLR1 gene, an Ikeda A, Ueguchi-Tanaka M, Sonoda Y, Kitano H, Koshioka M, Futsuhara Y,
Matsuoka M, Yamaguchi J (2001) Slender rice, a constitutive gibberellin
response mutant, is caused by a null mutation of the SLR1 gene, an
|
https://openalex.org/W2512102792
|
https://figshare.com/ndownloader/files/10263381
|
English
| null |
Phase-Amplitude Separation and Modeling of Spherical Trajectories
|
Journal of computational and graphical statistics
| 2,018
|
cc-by
| 2,395
|
Appendix to “Phase-Amplitude Separation
and Modeling of Spherical Trajectories” 1. The Riemannian structure on S2. 2. Some properties for geodesics on C. 3. The proofs of Lemma 2 and Theorem 1. 4. The derivation of exponential map and inverse exponential map on C. 5. The comparison of two modeling strategies in Section 3.1. 1
Riemannian Structure on S2 To perform the trajectory analysis on the manifold S2, one needs a Riemannian structure on
this manifold. Specially, we need the following tools: (1) geodesic between two points on the
manifold, (2) parallel transport of tangent vectors along the geodesic path, (3) exponential
map, (4) inverse exponential map and (5) Riemannian curvature tensor. We use a simple Euclidean inner product as the Riemannian metric on S2: for any
v1, v2 ∈Tp(M), the metric is defined to be: ⟨v1, v2⟩= vt
1v2. For any two points p, q ∈ 1 S2(p ̸= q) and a tangent vector v ∈Tp(S2), we have the following closed solutions for the
tools we need: S2(p ̸= q) and a tangent vector v ∈Tp(S2), we have the following closed solutions for the
tools we need: 1. Geodesic: The geodesic between p and q is the great circle connecting them: α(t) =
1/sin(θ)(sin(θ(1 −t))p + sin(θt)q), where θ is determined by cos(θ) = ⟨p, q⟩and 0 <
θ < π 1. Geodesic: The geodesic between p and q is the great circle connecting them: α(t) =
1/sin(θ)(sin(θ(1 −t))p + sin(θt)q), where θ is determined by cos(θ) = ⟨p, q⟩and 0 < 1. Geodesic: The geodesic between p and q is the great circle connecting them: α(t) =
1/sin(θ)(sin(θ(1 −t))p + sin(θt)q), where θ is determined by cos(θ) = ⟨p, q⟩and 0 < 2. Parallel Transport: The parallel transport (v)p→q along the shortest geodesic (i.e. great circle) from p to q is given by v −2 ⟨v, q⟩(p + q)/∥p + q∥2. 3. Exponential Map: The exponential map expp(v) is cos(∥v∥)p + sin(∥v∥)v/∥v∥. 3. Exponential Map: The exponential map expp(v) is cos(∥v∥)p + sin(∥v∥)v/∥v∥. 4. Inverse Exponential Map: The inverse exponential map exp−1
p (q) is (q−p cos(θ))θ/ sin(θ),
θ = cos−1(⟨g1, g2⟩). 5. Riemannian Curvature Tensor: For three tangent vectors x, y, z on Tp(S2), the Rie-
mannian curvature tensor R(x, y)(z) = ⟨y, z⟩x −⟨x, z⟩y = −(x × y × z), where ⟨·⟩
denotes the ordinary inner product, and × denotes the cross product. 5. Riemannian Curvature Tensor: For three tangent vectors x, y, z on Tp(S2), the Rie-
mannian curvature tensor R(x, y)(z) = ⟨y, z⟩x −⟨x, z⟩y = −(x × y × z), where ⟨·⟩
denotes the ordinary inner product, and × denotes the cross product. 3
Proof of Lemma 2 Let β be the shortest geodesic joining p1 to p2 and let L : Tp1(S2) →Tp2(S2) be the parallel
translation map induced by β. Let ζ be any other path from p1 to p2 that is disjoint from
β. The Gauss Bonnet Theorem states that the angle of rotation of the parallel translation
map Tp2(S2) →Tp2(S2) induced by the concatenation ζ−1 ∗β is equal to the integral of
the Gaussian curvature over the region enclosed by the loop β ∪ζ. Since the Gaussian
curvature of S2 equals +1 at every point, this implies that this angle of rotation is equal to
the area enclosed by the loop. However, it is well known that of all curves that enclose a
given area, a circle is the shortest! From this, it is easy to prove that if part of your loop
is already given (by the geodesic, as in this case), then the shortest way to fill in the rest
of your arc to enclose a given area is by a circular arc. This proves Lemma 2. 2
Two properties for geodesics on C Suppose that (β(t), q(t)) is a geodesic on C, where t ∈I = [0, 1]. Let L denote the length
of the path β : I →S2. Let ˜β : I →S2 be a constant speed re-parametrization of β. For
each t0 ∈I, define Zt0 : L2(I, T˜β(0)(S2)) →L2(I, T˜β(t0)(S2)) by parallel transporting each
tangent vector along ˜β(t). Then we can define Z : [0, L] × L2(I, T˜β(0)(S2)) →C by Z(s, q) = (˜β(s/L), Z(s/L)(q)). A routine verification shows that if we put the standard
product Riemannian metric on [0, L] × L2(I, T˜β(0)(S2)), then Z is an isometric immersion. Since our original geodesic (β(t), q(t)) is contained in the image of Z, its inverse image under
Z in [0, L] × L2(I, T˜β(0)(S2)) must be a geodesic. But since this latter space is Euclidean
(i.e., we are using the same Riemannian metric at each point), it follows that the inverse
image of our geodesic must be a straight line in this space. It follows immediately that
β(t) must have constant speed and q(t) must be covariantly linear. 2 4
Proof of Theorem 1 Let γ ∈Γ be a warping function, and let γ act on the space C by (x, q)∗γ = (x, (q∗γ)). The
differential of this action is the map T(x,q)(C) →T(x,(q∗γ))(C) given by (u, w) 7→(u, w ∗γ). We prove that this differential preserves our Riemannian inner product (Eqn. 1) as follows:
let (u1, w1) and (u2, w2) be two tangent vectors on T(x,q)(C); it follows that ⟨(u1, w1 ∗γ), (u2, w2 ∗γ)⟩
=
u1 · u2 +
∫1
0
w1(γ(t))
√
˙γ(t)w2(γ(t))
√
˙γ(t)dt
=
u1 · u2 +
∫1
0
w1(γ)w2(γ)dγ
=
u1 · u2 +
∫1
0
w1(s)w2(s)ds
=
⟨(u1, w1), (u2, w2)⟩ Since Γ acts on C by isometries, i.e. preserving the Riemannian inner product, it follows
immediately that it takes geodesics to geodesics, and preserves geodesic distance. It also
follows that it preserves the baselines of these geodesics, i.e. β∗. Since Γ acts on C by isometries, i.e. preserving the Riemannian inner product, it follows
immediately that it takes geodesics to geodesics, and preserves geodesic distance. It also
follows that it preserves the baselines of these geodesics, i.e. β∗. 3 5
Exponential and Inverse Exponential Map on C 5 In this section, we will define the exponential map and inverse exponential map on C . Inverse Exponential Map on C: Given (p1, q1) and (p2, q2), let {(β(s), q(s, ·))|s ∈[0, 1]}
be the geodesic connecting them on C. The inverse exponential map from (p2, q2) to (p1, q1)
is defined to be the mapping from C to T(p1,q1)(C) such that exp−1
(p1,q1)(p2, q2) = (u1, w1),
where u1 ∈Tp1(S2) and u1 ⊥p1 with ∥u1∥= lβ. The expressions for u1 and w1 are given
below: • u1 = lβ
Aθp1
∥Aθp1∥, where Aθ is defined in Algorithm 2 such that β(s) = esAθp1. It is easy
to show that Aθ is an asymmetric matrix, and trace(Aθ) = 0. • u1 = lβ
Aθp1
∥Aθp1∥, where Aθ is defined in Algorithm 2 such that β(s) = esAθp1. It is easy
to show that Aθ is an asymmetric matrix, and trace(Aθ) = 0. • w1 = q∥
2,β −q1 ∈L2([0, 1], Tp1(S2)), where q∥
2,β denotes the backward parallel transport
of q2 along β from p2 to p1. Exponential Map on C: Given a point (p1, q1) in C and a tangent vector (u1, w1) ∈
T(p1,q1)(C), the exponential map is a mapping from T(p1,q1)(C) to C. Furthermore, exp(p1,q1)(s(u1
with s ∈[0, 1], provides a geodesic path on C; we will denote it by (β(s), q(s, ·)). Since the
geodesic is determined by the baseline β, we only need to find an asymmetric matrix A
as described in Algorithm 2 (in the main paper) such that esAp1 = β(s) to determine the
baseline. Here, A has three unknown parameters because trace(A) = 0. To solve for A, we
have the following equations: 1. The vector fields ˙β(s) take the value u1 at s = 0 by definition, and we have ˙β(s)|s=0 =
Ap1. Therefore, the first equation for solving A is Ap1 = u1. (1) (1) 2. According to the geodesic equations derived in Zhang et al. (2015) on C, the second
derivative of the baseline β is determined by: 2. According to the geodesic equations derived in Zhang et al. (2015) on C, the second
derivative of the baseline β is determined by: 2. According to the geodesic equations derived in Zhang et al. 5
Exponential and Inverse Exponential Map on C (2015) on C, the second
derivative of the baseline β is determined by: ∇˙β(s)( ˙β(s)) = −R(q(s), (∇˙β(s)q)(s)) ˙β(s), ∇˙β(s)( ˙β(s)) = −R(q(s), (∇˙β(s)q)(s)) ˙β(s), where R(·, ·) denotes the Riemannian curvature tensor. In the left side, we have
∇˙β(s)( ˙β(s))|s=0 = PTp1(S2)
(
d2
ds2esAp1|s=0
)
, where PTp1(S2)(·) denotes the projection of where R(·, ·) denotes the Riemannian curvature tensor. In the left side, we have
∇˙β(s)( ˙β(s))|s=0 = PTp1(S2)
(
d2
ds2esAp1|s=0
)
, where PTp1(S2)(·) denotes the projection of 4 a vector to the tangent space Tp1(S2). In the right side, given the baseline β, we
know that q(s, ·) = (q1 + sw1)p1→β(s) = (q1)p1→β(s) + s(w1)p1→β(s). Therefore, we
have (∇˙β(s)q)(s) = (w1)p1→β(s). At the point s = 0, R(q(s), (∇˙β(s)q)(s)) ˙β(s)|s=0 =
R(q1, w1) ˙β(0). Therefore, the above equation simplifies at s = 0 to become: PTp1(S2)
( d2
ds2esAp1
)
= −R(q1, w1)u1 . (2) (2) (2) (1) and (2) are used to solve for the asymmetric matrix A as the function of p1, q1, u1
and w1. (1) provides two equations to solve unknown parameters in A since u1 ⊥p1. It is
the same for (2) which provides two additional equations. So there are four equations for
three unknown parameters in A and one is redundant. Given A, the exponential map can
be expressed as exp(p1,q1)(s(u1, w1)) = (esAp1, (q1 + sw1)p1→esAp1). 6
Comparison of Two PCA Modeling Strategies In this section, we compare the two PCA modeling strategy proposed in Section 3.1. While
the first model treats the starting point and the velocity curve separately, the second one
models them jointly. We perform the comparison on both the bird migration and hurricane
track data. Separate Modeling: We calculate and show the first two principal directions for the
two components separately in Fig. 1. In each row, in the left panel, we let the second
component w = 0 and show the two modes of variation in the first component ui, and in
the right panel, we let u = 0 and show the first two modes of variation in wi. The middle
curve in magenta color, with τ = 0, is the mean trajectory. In the parentheses in each row,
we show the percentage of variation that was explained by the first two PCs. Joint Modeling: To better characterize the dependence between the starting point
and velocity curve, we jointly model them. Let U be the data matrix with i-th row being
U(i, :) = (sut
i, wt
i). The covariance matrix KU is calculated. Here we set s =
√
T/10 to
adjust for the effect of using different number of sample points T for the second component
wi. The first two principal directions estimated from the joint model are shown in Fig. 2. From the percentage of variation explained by the first two PCs, we do not see much
improvement after jointly modeling, but we do obtain significantly different PCs comparing 5 First two PCs for u with w = 0 (100%)
First two PCs for w with u = 0 (41.89%)
First two PCs for u with w = 0 (100%)
First two PCs for w with u = 0 (69.44%)
Figure 1: Separate modeling result: PCA for bird migration (first row) and hurricane data
(second row). 6
Comparison of Two PCA Modeling Strategies First two PCs for u with w = 0 (100%) First two PCs for u with w = 0 (100%) First two PCs for w with u = 0 (41.89%) (
)
First two PCs for w with u = 0 (69 44%) First two PCs for w with u = 0 (69.44%) First two PCs for u with w = 0 (100%) First two PCs for w with u = 0 (69.44%) First two PCs for u with w = 0 (100%) Figure 1: Separate modeling result: PCA for bird migration (first row) and hurricane data
(second row). with separate modeling method (due to the combination of the starting point and the
velocity function). Certainly, we would like to see if the joint modeling helps us to better characterize the
variation among the data. We use the wrapped models described in Section 3.2 to generate
random samples after fitting a distribution of the observed data. Fig. 3 displays some
random samples from the estimated parametric Gaussian distributions (for two components
separately). From this result, we discover that the variance in re-sampled trajectories
seems larger than the original ones, which indicates that independently modeling the two
components using Gaussian distribution is not a perfect solution. In Fig. 4 we show the
sampling results of the joint modeling strategy. We assume that the data follow a mixture
K Gaussian distributions. From the random sampling result, we see that the joint modeling
strategy together with the mixture distribution give us a better characterization of observed
data. To select K, we suggest to use AIC scores. 6 First two PCs for bird data (41.90%)
First two PCs for hurricane data (69.43%)
Figure 2: Joint modeling result: PCA for bird migration (left) and hurricane data (right). First two PCs for hurricane data (69.43%) First two PCs for hurricane data (69.43%) First two PCs for bird data (41.90%) Figure 2: Joint modeling result: PCA for bird migration (left) and hurricane data (right). Figure 3: Examples of random sampling from the parametric Gaussian distributions (in
separate modeling scenario). Figure 3: Examples of random sampling from the parametric Gaussian distributions (in
separate modeling scenario). Figure 3: Examples of random sampling from the parametric Gaussian distributions (in
separate modeling scenario). (a) K = 1
(b) K = 2
(c) K = 1
Figure 4: Examples of random sampling from K mixture Gaussian distributions (in joint
modeling scenario). 6
Comparison of Two PCA Modeling Strategies Figure 4: Examples of random sampling from K mixture Gaussian distributions (in joint
modeling scenario). 7 7 References Zhang, Z., Su, J., Klassen, E., Le, H. & Srivastava, A. (2015), ‘Video-based action recog-
nition using rate-invariant analysis of covariance trajectories’, arXiv 1503.06699. 8 8
|
https://openalex.org/W4229556947
|
https://www.qeios.com/read/U7NU8Y/pdf
|
English
| null |
Recombinant Growth Factor
|
Definitions
| 2,020
|
cc-by
| 80
|
Qeios · Definition, February 7, 2020 Open Peer Review on Qeios Open Peer Review on Qeios Recombinant Growth Factor National Cancer Institute National Cancer Institute Qeios ID: U7NU8Y · https://doi.org/10.32388/U7NU8Y Source National Cancer Institute. Recombinant Growth Factor. NCI Thesaurus. Code C530. Formulated therapeutic proteins that are either isolated from endogenous sources or
manufactured in a laboratory. Growth factors bind to specific receptors expressed by
various cell types, thereby stimulating cell differentiation and/or proliferation. (NCI04) Qeios ID: U7NU8Y · https://doi.org/10.32388/U7NU8Y 1/1
|
https://openalex.org/W3045296427
|
https://link.springer.com/content/pdf/10.1007%2F978-3-662-61755-7_16.pdf
|
English
| null |
Measurement based robot cutting as preparation for welding tubular connections
|
Springer eBooks
| 2,020
|
cc-by
| 3,610
|
Measurement based robot cutting as preparation for
welding tubular connections Sascha Lauer1, Tina Ambrosat1, Martin-Christoph Wanner1, Wilko Flügge1,2 1Fraunhofer Research Institution of Large Structures in Production Engineering IGP, Albert-
Einstein-Straße 30, 18059 Rostock, Germany
2Chair of Production Technology, University of Rostock, Albert-Einstein-Straße 2, 18059 Ros-
tock, Germany
{sascha.lauer, tina.ambrosat, martin-christoph.wanner,
wilko.fluegge}@igp.fraunhofer.de 1Fraunhofer Research Institution of Large Structures in Production Engineering IGP, Albert-
Einstein-Straße 30, 18059 Rostock, Germany
2Chair of Production Technology, University of Rostock, Albert-Einstein-Straße 2, 18059 Ros-
tock, Germany
{sascha.lauer, tina.ambrosat, martin-christoph.wanner,
wilko.fluegge}@igp.fraunhofer.de {sascha.lauer, tina.ambrosat, martin-christoph.wanner,
wilko.fluegge}@igp.fraunhofer.de Abstract. In the production of offshore tubular connections, the cutting process
is an important factor. Undefined root gaps between the individual pipe connec-
tions require time-consuming reworking and increased welding effort, resulting
in longer production times and higher costs. The developed procedure is based
on the 3D measurement of the pipes in order to consider the real geometries in
the planning of the cutting process and to optimize the following joining process
by adapting the root gap all over the tubular connection and to get a constant
seam cross section. The basic concept is to build the planned connection virtually
with the measured and real pipe geometries and to plan the flame cutting process
including weld seam preparation on the base of the result. For the measurement
in the production environment, a suitable measuring concept was developed to
return the results directly into the cutting process with the robot. This is the base
for the automation of the tubular connection production. Keywords: offshore structures, sensor-based programming, path planning, ro-
botic cutting © The Editor(s) (if applicable) and The Author(s) 2020
T. Schüppstuhl et al. (Hrsg.), Annals of Scientific Society for Assembly,
Handling and Industrial Robotics, https://doi.org/10.1007/978-3-662-61755-7_16 1
Introduction Jacket structures play an important role in offshore technology. They are used as foun-
dation for converter platforms at sea as well as for wind energy plants. Jackets are com-
plex tubular connections based on pipes (Fig. 1). Compared to other offshore founda-
tion structures, the used pipes have smaller diameters and material thicknesses. Accord-
ingly, jackets can be produced with lower material costs than comparable structures
with similar functions, such as monopiles. However, the complexity results in higher
production costs. An essential cost driver are the deviations in the pipe production, re-
sulting with time-consuming and cost-intensive manual rework in the assembly. © The Editor(s) (if applicable) and The Author(s) 2020
T. Schüppstuhl et al. (Hrsg.), Annals of Scientific Society for Assembly,
Handling and Industrial Robotics, https://doi.org/10.1007/978-3-662-61755-7_16 174 Fig. 1. Jacket structure (left), tubular connection (right) [1] Fig. 1. Jacket structure (left), tubular connection (right) [1] The main reason for the need for reworking was identified as the determination of the
cutting contours without taking into account the production deviations of the real pipes. By measuring the real tube geometries and by not using the CAD geometry, the manu-
facturing process for joining the tubular connections is to be adapted in order to mini-
mize manual reworking and thus reduce manufacturing costs. Fig. 2 shows the manufacturing process of a tubular connection as a process chain. The
process adapts the cutting process to the tolerances in pipe production and serves as the
b
i f
iti
i
d j i i
th
i basis for positioning and joining the pipe. Fig. 2. Process chain for tubular connection production Fig. 2. Process chain for tubular connection production Tubular connections consist of a main member pipe and one or more branch member
pipes (see Fig. 3), whereby the pipe diameters used here varied from 500 mm to
3000 mm and the material thicknesses between 20 mm and 120 mm. Tubular connections consist of a main member pipe and one or more branch member
pipes (see Fig. 3), whereby the pipe diameters used here varied from 500 mm to
3000 mm and the material thicknesses between 20 mm and 120 mm. By flame cutting, the pipe of the branch member is prepared for the joining process. The contour which is calculated by virtually placing the branch member pipe on the
main member pipe is based on the ideal geometries of both pipes. 1
Introduction In addition, the weld
seam preparation is planned for the branch member pipe by using this contour. For
conforming the standards of joining for this tubular connection it requires a defined root
gap between 2 and 8 mm [2], which depends on the different zones over the various
areas of the weld (see I, II and III in Fig. 3). 175 Fig. 3. K-connection with definitions of root gap, dihedral angle (opening angle between sur-
faces) and the zones (I – heel zone; II – side zone; III – toe zone) Fig. 3. K-connection with definitions of root gap, dihedral angle (opening angle between sur
faces) and the zones (I – heel zone; II – side zone; III – toe zone) Due to the production deviation of both pipes, the ideal seam course in the pipe joint
must be adapted to reality. If this is not the case, the joining technology will not be able
to tolerate root gaps, which will lead to serious manual reworking such as grinding of
the weld in the cut surface. The conventional manufacturing of tubular connections
doesn’t consider the real geometries of the main member pipe during weld seam prep-
aration. Only the real pipe geometry of the branch member is used to ensure the quality
of the cut. By planning the contour based on real pipe geometries and by generating constant
seam cross sections in the weld seam preparation, automated welding with the specially
developed orbital welding system in high quality is possible [3]. 2
Description of the developed process In order to achieve these goals, the manufacturing process has to be adapted and the
measurement of the pipes integrated into it. Specifically, the following steps (Fig. 4)
are necessary:
Recording the geometry of the main and the branch member
Determination of the virtual cutting contour
Inclusion of weld seam preparation
Flame cutting of the pipe with a robot Compared to the state of the art, the first two steps reduce the expensive and timecon-
suming re-working by a slightly increasing production planning. 176 Fig. 4. Process steps for cutting with robots
1. digitization
3. seam preparation
4. cutting process
2. cutting contour Fig. 4. Process steps for cutting with robots In the following, the mentioned four steps are examined using an example tubular
connection. In the selected configuration, a branch member pipe with a diameter of
508 mm and a material thickness of 20 mm is placed centrally at an angle of 45° on the
main member pipe with a diameter of 1016 mm. 2.1
Test setup For the execution of tests, a suitable plant design was developed. A welding robot
(KUKA kr5 arc) in combination with a turn and tilt positioner (KUKA DKP400) is
used to extend the working area of the robot. The positioner can handle pipes up to a
mass of 400 kg, which is necessary for the chosen branch member pipe. Fig. 5. Test setup for the robot cutting Fig. 5. Test setup for the robot cutting 177 In order to work with the highest accuracy during the entire process, the coordinate
systems of the tilt and turn positioner and the robot were measured to each other using
an external measuring system (laser tracker). After the robot system was measured, it
was referenced to a selected global system defined by reference marks. This is used
throughout the process. 2.2
Digitization of the pipes Both pipes are digitized in the production environment by a laser scanner. The used
Z+F IMAGER® 5010 has a linearity error of less than 1 mm and a distance noise of
0.5 mm rms at 10 meter [4]. Distances of more than 10 meter to the measuring object
are not provided for in the measuring concept. The main member pipe is measured horizontally. The coordinate system is defined by
the pipe axis, flange plane and a marking of the theoretical intersection point of the pipe
axis with the pipe surface. When measuring the branch member pipe, it is perpendicular to the tilt and turn posi-
tioner. The robot has a pose repeatability of ± 0.06 mm [5], which is used throughout
the process. Instead of measuring the tube from different angles and positions, only one
laser scanner position is required and the pipe is rotated between each scan using the
tilt and turn positioner (see Fig. 6). The registration of the scans takes place in the co-
ordinate system of the tilt and turn positioner, defined by the rotary axis of the table
and a rotary angle. The generated point clouds of the outer surface of the branch mem-
ber pipe are assembled in the evaluation software by turning them back about the de-
fined rotary angle of the positioner. Due to the overlapping areas of the various scans
and the unfavorable impact angle of the laser, data smoothing is necessary. p
g
g
y
Fig. 6. Measuring setup for the measurement of the branch member pipe Fig. 6. Measuring setup for the measurement of the branch member pipe 178 A measurement of the branch member pipe from the inside does not take place, since
the effort seems inappropriate due to the inner diameter and difficult referencing. Dur-
ing preparatory measurements of the pipe geometries, it was determined that the mate-
rial thicknesses of the pipes, with the exception of the weld seam, is approximately
constant. Therefore, the data points of the inner pipe surface are simulated on the base
of the data points of the outer pipe surface using the pipe axis determined by cylinder
segmentation according to [6]. The planned configuration is now virtually simulated
with the point clouds of the main member pipe and the branch member. 2.2
Digitization of the pipes The axis of
branch member pipe is transformed to the z-axis and the axis of the main member pipe
is aligned at the defined angle (see Fig. 7 (left)). The breakthrough point of the z-axis
on the surface of the main member pipe is known from the measurement concept. By
translating the point cloud parallel to the pipe axis, the dimensions of the future socket
are virtually taken into account. are virtually taken into account. Fig. 7. Process flow for determining the virtual intersection contour using the example of the
outer surface of the extension tube, virtual alignment (left), polygonised intersection areas (cen-
ter) and intersection contour (right) Fig. 7. Process flow for determining the virtual intersection contour using the example of the
outer surface of the extension tube, virtual alignment (left), polygonised intersection areas (cen-
ter) and intersection contour (right) Using trigonometric methods, the intersection areas of the point clouds were identi-
fied on the base of the configuration and the points lying within them were polygonised
(see Fig. 7 (center)). From the calculation of the section of the polygonised areas, a 3D
polyline is obtained, with a C³ continuous, closed BSpline curve approximated by its
straight line center points (see Fig. 7 (right)). The results are the curves (inner and outer
contour), in which point cloud of the main member pipe crosses the point cloud of the
branch member pipe. The outer curve corresponds to the seam contour (see Fig. 3), on
which the weld seam preparation takes place. 2.3
Weld seam preparation The dihedral angle is used to determine the seam course [2]. Fig. 8 shows a seam course
corresponding to the standard for the production of a pipe joint with the used configu-
ration. If the dihedral angle is smaller than 50°, a fillet weld is prepared (zone I, Fig. 3),
if it is larger than 90°, a bevel groove is prepared (zone III, Fig. 3). The area in between
is called the transition area (zone II, Fig. 3), because the change between these two
seam types takes place. The course of this seam was defined in such a way that it can
be flame cut with a single move. The challenge is that the seam opening angles which
needs to be cut becomes acute. For later use, this step has to be adapted to a two-part
cut. 179 Fig. 8. Seam definition in the selected configuration, depending on the dihedral angle Fig. 8. Seam definition in the selected configuration, depending on the dihedral angle The shown dihedral angles were determined with the seam contour of an ideal tub-
ular connection with faultlessly pipes. In case of a fillet weld, the main member pipe
and the branch member pipe touch each other at the outer contour. In case of a bevel
groove, the inner contour of the branch member pipe is in contact. Fig. 9. Target layer structure for tubular connection at different areas (branch member – top,
main member – bottom) Fig. 9. Target layer structure for tubular connection at different areas (branch member – top,
main member – bottom) The necessity of an accurately planned cut and the contact between both pipes is
shown in Fig. 9. For a future automated welding process, with the aim of achieving a
uniform layer structure, it is essential to achieve a constant root gap and a defined weld
cross section. Otherwise, the welding parameter has to be adjusted during the entire
joining process. 2.4
Offline programming and Preparation for flame cutting Since the flame cut is placed on the outer surface of the branch member pipe, the cal-
culated points for the robot have to be shifted to the outer surface of the branch member 180 pipe along the cutting direction (see Fig. 10 (left)). For a constant cutting quality, the
torch distance to the cutting surface has to be constant. This is achieved by further
shifting each point along the corresponding cutting direction by an offset. A retransfor-
mation of the data provides the discrete positions and directions of the torch head in the
coordinate system of the tilt and turn positioner. The robot is integrated in this coordi-
nate system. Since the tilt-turn positioner has a higher accuracy than the robot, the turn-
table rotates during the cutting process, while the robot only performs small move-
ments. The robot software interpolates between the position and direction data and cal-
culates the positions of the axis in order to avoid singularity positions. By using syn-
chronized movements from the positioner and the robot, it is possible to cut at constant
speed using all eight axes. p
g
g
Fig. 10. Calculated contour (left) and path planning with the robot in SIEMENS RobotExpert
(right) Fig. 10. Calculated contour (left) and path planning with the robot in SIEMENS RobotExper
(right) This is the base for the offline-programming of robot motion (see Fig. 10, right) and
collision control to prevent crashes between the robot/torch and the pipe. In addition,
motion commands and switching commands for the cutting process, such as start and
stop distances, ignition commands and waiting times, have to be added. 3
Verification and Results 3 The method is used to manufacture the described tubular connection. The cut branch
member pipe, especially the cut surface, is measured with the strip light projection sys-
tem GOM Atos Triple Scan. A 300 x 300 mm² measuring field is used, with which a
measuring accuracy of approx. 0.01 mm can be achieved [7]. First of all the resulting model is compared with an ideal cylinder to determine the
production deviations. On the other hand, the model is virtually placed on the digitized
main member pipe in the defined configuration and aligned by best-fit method, whereby
only rotation around and displacement along the pipe axis are permitted. The root gap
can be derived from this. The results are shown in the form of a measurement in Fig. 11. 181 Fig. 11. Measurement results to illustrate the production deviations of an extension tube (left)
and to illustrate the deviation of the cut surface from the planned one (right). Fig. 11. Measurement results to illustrate the production deviations of an extension tube (left)
and to illustrate the deviation of the cut surface from the planned one (right). The production deviations of the attachment pipe are clearly visible in the left meas-
urement. The tolerance of ±5 mm is maintained [8]. The right measurement shows the
deviations between the virtually planned and the actual cut surface. The maximum de-
viation, this means the largest gap between both pipes, is 1.59 mm. The minimum de-
viation, the largest penetration of the two pipes, is -1.83 mm. They vary by 3.42 mm. Within the bevel groove area (zone III, Fig. 3, upper cut area, Fig. 11) the deviations
differ by less than 1 mm. Within the fillet weld area (zone I, Fig. 3, lower cut area,
Fig. 11), too. The variations in the transition areas are more than 2 mm. The specified
fluctuations of the nominal root gap of 6 mm are adhered to. Despite the production
deviations, the weld seam can be prepared in accordance with the standards. References 1. Brauser, S, Lauer, S.: Mechanisierung eines Mehrlagenschweißprozesses zum Fügen von
Offshore-Rohrverbindungen, 8. Zukunftskonferenz: Wind & Maritim, Rostock 2019 1. Brauser, S, Lauer, S.: Mechanisierung eines Mehrlagenschweißprozesses zum Fügen von
Offshore-Rohrverbindungen, 8. Zukunftskonferenz: Wind & Maritim, Rostock 2019 g
2. American Welding Society: AWS D1.1:Structural Welding Code-Steel, Miami (2000) 3. Herholz, H., Dryba, S., Gründler, M., Wanner, M.-C.: Development of a flexible special
kinematics for fully mechanized production of tubular nodes for offshore foundation struc-
tures. In: Tagungsband des 2. Kongresses Montage Handhabung Industrieroboter, pp. 1-11. Springer, Heidelberg (2017) 4. Zoller + Fröhlich GmbH, Z+F IMAGER 5010 Datenblatt, Bridgewille:Z+F USA 4. Zoller + Fröhlich GmbH, Z+F IMAGER 5010 Datenblatt, Bridgewille:Z+F USA
5. KUKA Roboter GmbH, KUKA Positionierer DKP-400, DKP-400 endlos Spezifikation,
Augsburg 2013 5. KUKA Roboter GmbH, KUKA Positionierer DKP-400, DKP-400 endlos Spezifikation,
Augsburg 2013 6. Rusu, R. B., Cousins,S.: 3D is here: Point Cloud Library (PCL), Menlo Park 2011 7. GOM GmbH, Atos Triple Scan – Industrieller optischer 3D-Digitalisierer, Braunschwe
2017 8. Deutsches Institut für Normung: DIN EN 10216-1:2014-03 Nahtlose Stahlrohre für Druck-
beanspruchungen – Technische Lieferbedingungen – Teil 1:Rohre aus unlegierten Stählen
mit festgelegten Eigenschaften bei Raumtemperatur, Beuth Verlag 2014
9. Ambrosat,T., Lauer,S., Geist,M. Flügge,W.:Bestimmung komplexer Schneidkonturen als
Vorbereitung zum Verschweißen von 3D Rohrstößen GFaI Berlin 2018 8. Deutsches Institut für Normung: DIN EN 10216-1:2014-03 Nahtlose Stahlrohre für Druck-
beanspruchungen – Technische Lieferbedingungen – Teil 1:Rohre aus unlegierten Stählen
mit festgelegten Eigenschaften bei Raumtemperatur, Beuth Verlag 2014 8. Deutsches Institut für Normung: DIN EN 10216-1:2014-03 Nahtlose Stahlrohre für Druck-
beanspruchungen – Technische Lieferbedingungen – Teil 1:Rohre aus unlegierten Stählen
mit festgelegten Eigenschaften bei Raumtemperatur, Beuth Verlag 2014 9. Ambrosat,T., Lauer,S., Geist,M. Flügge,W.:Bestimmung komplexer Schneidkonturen als
Vorbereitung zum Verschweißen von 3D Rohrstößen, GFaI, Berlin 2018 9. Ambrosat,T., Lauer,S., Geist,M. Flügge,W.:Bestimmung komplexer Schneidkonturen als
Vorbereitung zum Verschweißen von 3D Rohrstößen, GFaI, Berlin 2018 Open Access This chapter is licensed under the terms of the Creative Commons Attri-
permits use, sharing, adaptation, distribution and reproduction in any medium or for-
mat, as long as you give appropriate credit to the original author(s) and the source,
provide a link to the Creative Commons license and indicate if changes were made. bution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which The images or other third party material in this chapter are included in the chap-
ter’s Creative Commons license, unless indicated otherwise in a credit line to the
material. 4
Conclusion and outlook It has been proven that the interaction of measurement technology, cutting contour
planning and robot cutting can be successfully applied in practice. Both the tubular
connection configuration and the selected seam configuration are to be understood as
an example. The process can be used for any tubular connection and seam configura-
tion. Further optimization possibilities are in the next process steps of positioning and
joining (see Fig. 2). The challenge in positioning is that the approach point planned in
the configuration, in which the branch member pipe axis meets the axis of the main
member pipe, is calculated and has to be transferred to the real pipe configuration. In order to optimize the joining process, based on a two-part cut, a suitable seam con-
figuration can be developed. Because the seam course is planned and implemented on
the base of the actual data, virtual weld layer planning is possible without further meas-
urement, which additionally accelerates the production process. In particular, this ena-
bles automation of the welding process. It has been proven that the interaction of measurement technology, cutting contour
planning and robot cutting can be successfully applied in practice. Both the tubular
connection configuration and the selected seam configuration are to be understood as
an example. The process can be used for any tubular connection and seam configura-
tion. Further optimization possibilities are in the next process steps of positioning and
joining (see Fig. 2). The challenge in positioning is that the approach point planned in
the configuration, in which the branch member pipe axis meets the axis of the main
member pipe, is calculated and has to be transferred to the real pipe configuration. In order to optimize the joining process, based on a two-part cut, a suitable seam con-
figuration can be developed. Because the seam course is planned and implemented on
the base of the actual data, virtual weld layer planning is possible without further meas-
urement, which additionally accelerates the production process. In particular, this ena-
bles automation of the welding process. 182 References If material is not included in the chapter’s Creative Commons license and
your intended use is not permitted by statutory regulation or exceeds the permitted
use, you will need to obtain permission directly from the copyright holder.
|
https://openalex.org/W2912583419
|
https://europepmc.org/articles/pmc6393874?pdf=render
|
English
| null |
Relation of Redox and Structural Alterations of Rat Skin in the Function of Chronological Aging
|
Oxidative medicine and cellular longevity
| 2,019
|
cc-by
| 9,961
|
Aleksandra Jankovic
,1 Luciano Saso
,2 Aleksandra Korac
,3 and Bato Korac
1,3
1Institute for Biological Research “Sinisa Stankovic”, University of Belgrade, Serbia
2Department of Physiology and Pharmacology “Vittorio Erspamer”, Faculty of Pharmacy and Medicine,
S
i
U i
it
f R
It l Aleksandra Jankovic
,1 Luciano Saso
,2 Aleksandra Korac
,3 and Bato Korac
1,3
1Institute for Biological Research “Sinisa Stankovic”, University of Belgrade, Serbia
2Department of Physiology and Pharmacology “Vittorio Erspamer”, Faculty of Pharmacy and Medicine,
Sapienza University of Rome Italy 1Institute for Biological Research “Sinisa Stankovic”, University of Belgrade, Serbia
2Department of Physiology and Pharmacology “Vittorio Erspamer”, Faculty of Pharmacy and Medicine,
Sapienza University of Rome, Italy 1Institute for Biological Research “Sinisa Stankovic”, University of Belgrade, Serbia
2Department of Physiology and Pharmacology “Vittorio Erspamer”, Faculty of Pharmacy and Medicine,
Sapienza University of Rome, Italy 3Faculty of Biology, University of Belgrade, Serbia Correspondence should be addressed to Bato Korac; koracb@ibiss.bg.ac.rs Received 31 July 2018; Accepted 9 December 2018; Published 14 February 2019 Academic Editor: Ana Cipak Gasparovic Copyright © 2019 Aleksandra Jankovic et al. This is an open access article distributed under the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is
properly cited. Accumulation of oxidative insults on molecular and supramolecular levels could compromise renewal potency and architecture in
the aging skin. To examine and compare morphological and ultrastructural changes with redox alterations during chronological
skin aging, activities of antioxidant defense (AD) enzymes, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase
(GSH-Px), glutathione reductase (GR), thioredoxin reductase (TR), and methionine sulfoxide reductase A (MsrA), and the
markers of oxidative damage of biomolecules—4-hydroxynonenal (HNE) and 8-oxoguanine (8-oxoG)—were examined in the
rat skin during life (from 3 days to 21 months). As compared to adult 3-month-old skin, higher activities of CAT, GSH-Px, and
GR and a decline in expression of MsrA are found in 21-month-old skin. These changes correspond to degenerative changes at
structural and ultrastructural levels in epidermal and dermal compartments, low proliferation capacity, and higher levels of
HNE-modified protein aldehydes (particularly in basal lamina) and 8-oxoG positivity in nuclei and mitochondria in the
sebaceous glands and root sheath. In 3-day-old skin, higher activities of AD enzymes (SOD, CAT, GR, and TR) and MsrA
expression correspond to intensive postnatal development and proliferation. In contrast to 21-month-old skin, a high level of
HNE in young skin is not accompanied by 8-oxoG positivity or any morphological disturbances. Hindawi
Oxidative Medicine and Cellular Longevity
Volume 2019, Article ID 2471312, 12 pages
https://doi.org/10.1155/2019/2471312 Hindawi
Oxidative Medicine and Cellular Longevity
Volume 2019, Article ID 2471312, 12 pages
https://doi.org/10.1155/2019/2471312 Aleksandra Jankovic
,1 Luciano Saso
,2 Aleksandra Korac
,3 and Bato Korac
1,3
1Institute for Biological Research “Sinisa Stankovic”, University of Belgrade, Serbia
2Department of Physiology and Pharmacology “Vittorio Erspamer”, Faculty of Pharmacy and Medicine,
S
i
U i
it
f R
It l Observed results indicate that
increased activity of AD enzymes in elderly rat skin represents the compensatory response to accumulated oxidative damage of
DNA and proteins, accompanied by attenuated repair and proliferative capacity, but in young rats the redox changes are
necessary and inherent with processes which occur during postnatal skin development. Мorphological and ultrastructurаl
changes are in line with the redox profile in the skin of young and old rats. Oxidative Medicine and Cellular Longevity The inconsistency of the results is a conse-
quence of the different in vitro or in vivo studies, examining
different cell lines, tissues, and organs, parameters of oxida-
tive damage and antioxidative defense (AD), as well as the
period of the life cycle during which aging was examined. 2.2. Activity of Antioxidative Defense Enzymes: SOD, CAT,
GSH-Px, GR, and TR. Total specific SOD activity was deter-
mined by a modified method of Misra and Fridovich [22]. A degree of epinephrine autoxidation inhibition by SOD
contained in the examined samples in 0.05 M sodium
carbonate buffer (pH 10.2) was monitored spectrophotomet-
rically at 480 nm. The unit of SOD was defined as the amount
of enzyme that leads to 50% inhibition of the epinephrine
autoxidation rate under the appropriate reaction conditions
at 26°C. Catalase was assayed according to Beutler [23]. The
method is based on the rate of H2O2 degradation by the
action of catalase contained in the examined sample followed
spectrophotometrically at 230 nm in 5 mM EDTA and
Tris-HCl solution (pH 8.0). The catalase activity was calcu-
lated using a molar extinction coefficient for hydrogen perox-
ide (0.071), and values were expressed in units per mg of
protein. One unit of catalase was defined as the amount of
enzyme causing a decay of 1.0 μmol of H2O2 per minute at
25°C, at a 10mM hydrogen peroxide. To determine the activ-
ity of GSH-Px, a spectrophotometric method of Paglia and
Valentine [24] was used. The method is based on the mea-
surement of NADPH consumption (i.e., NADPH oxidation
by GR (Sigma, type III product) at 340 nm). This reaction is
preceded by the action of GSH-Px contained in the samples
examined on t-butyl hydroperoxide (3 mM) as a substrate
in 0.5 phosphate buffer (pH 7.0) at 37°C. The activity was
expressed as nmol NADPH oxidized min−1 mg−1 protein. GR activity was assayed by the method of Glatzle et al. [25]
and expressed as nmol NADPH min−1 mg−1 protein. This
procedure is based on GR-catalyzed reduction of oxidized
glutathione with NADPH, and oxidation of the latter was
determined spectrophotometrically at 340 nm, using 2 mM
GSSG and 0.1 mM NADPH, in phosphate buffer (pH 7.4),
at 37°C. The method of Luthman and Holmgren [26] was
used to determine TR activity. Oxidative Medicine and Cellular Longevity Oxidative Medicine and Cellular Longevity 2 “Siniša Stanković,” Belgrade. Healthy male Wistar rats were
kept in a temperature-controlled room (24 ± 1°C) on a
12/12 reverse light/dark cycle, with standard chow diet and
water ad libitum. Six individuals of the same age were housed
per cage. The rats were killed on the same day by decapitation
at different ages, from 3 days to 21 months. transfer the excitation energy to molecular oxygen, creating
singlet oxygen and other reactive species [6]. Also, the skin
contains a high level of polyunsaturated fatty acids in
membranes (25% of overall amount) and a high level of iron
(it excretes around 0.24-0.6 mg daily) that can additionally
increase oxidative pressure and reactive oxygen species
(ROS) production [7]. The skin was dissected within 3 min of death and thor-
oughly rinsed with physiological saline to remove traces of
blood. Parts of skin from the dorsal side were taken after
cutting the fur with an electric clipper. The hypodermis
was removed with a scalpel, and the remaining skin por-
tions were prepared for enzyme activity analysis and light
and electron microscopy. It is unequivocally demonstrated that ROS are involved
in many pathological conditions in the skin: immune sys-
tem disorders [8], vitiligo [9], psoriasis [10], acne and
ulcerations [11], rheumatoid arthritis [12], skin tumors
[13], and others. Moreover, when affected by exogenous
factors—drugs [14], UV radiation [15, 16], etc.—and
endogenous factors—inflammation [17], ischemia/reperfu-
sion [18], etc.—their production in the skin increases. In
addition,indifferentanimalmodels,morphologicalandstruc-
tural alterations in the epidermis and dermis of aged skin are
clearly shown. For analysis of AD enzyme activities, a portion of the tis-
sue weighing 1.5 g was homogenized (Ultra-Turrax homoge-
niser, Janke und Kunkel Ka/Werke, Staufen, Germany,
0-4°C) in 5 ml 0.25M sucrose, 0.1 mM EDTA, and 50mM
Tris-HCl buffer (pH 7.4). The homogenates were sonicated
and centrifuged as described previously [20]. Supernatants
were used for determination of protein concentration and
AD enzyme activities. The protein concentration in the
supernatant was estimated by the method of Lowry et al. [21]. However, the results of many studies examining the role
of free radicals in the aging process are inconsistent, often
contradictory. Oxidative Medicine and Cellular Longevity The method is based on the
use of TR as a catalyst for the NADPH-dependent reduction In previous decades, only one paper has been published
[19] which has comprehensively examined AD and markers
of oxidative damage in animals as a function of chronological
aging in female hairless mice aged 10 weeks (young) and 63
weeks (old). The authors conclude that neither the epidermis
nor the dermis showed changes in activity/amount of key
enzymatic and nonenzymatic components of AD, as well as
in the lipid hydroperoxide levels between young and old mice
(a histological comparison is absent in this study). In the current study, chronological aging in rat skin
was examined as a process inherent in life, in discrete time
intervals since birth (3rd day) to 21 months. In relation to
the function of aging, we have examined (i) the activities of
major antioxidant enzymes, superoxide dismutase (SOD),
catalase (CAT), glutathione peroxidase (GSH-Px), glutathi-
one reductase (GR), thioredoxin reductase (TR), and
methionine sulfoxide reductase A (MsrA) and (ii) the
markers of oxidative damage, 4-hydroxynonenal (HNE)
and 8-oxoguanine (8-oxoG). We have tried to establish two
additional imperatives in this study: firstly, to examine and
compare
morphological,
structural,
and
ultrastructural
changes with redox alterations during aging in skin and
secondly, to take a look at and discriminate all examined
parameters and processes during life (from 3 days to 21
months), comparing the changes in young (first two weeks
of birth, postnatal period when the skin intensively develops)
and old rats (rats aged 21 months) with those in adult
animals (3 months). 1. Introduction metabolism of fat (by forming depots) and in the metabo-
lism of water and salt [2]. Many theories try to explain aging, at least the molecular
basis of the pathophysiological processes (changes) that
accompany it. One of them is the free radical theory postu-
lated by Denham Harman [1], the universal theory that can
be applied to all types of cells and tissues, particularly to the
skin. The skin is one of the biggest organs in the body,
accounting for 16% of body weight, protects against
mechanical and radiation injuries and the entry of foreign
substances into the body, serves as a sensory organ rich in
nerves, regulates body temperature, and participates in the The skin is a mirror of aging [3]. Aging can be influenced
by endogenous (intrinsic aging) and exogenous (extrinsic
aging) factors. Oxidative pressure in skin which is exposed
to high partial pressure of oxygen from the inside (circulatory
system) and outside (50 times higher concentration) [4] can
be increased by activation of molecular oxygen by absorption
of light [5]. Other components in the skin (keratin proteins,
hemoglobin, porphyrins, carotene, nucleic acids, melanin,
lipoproteins, peptide bonds, and aromatic acids—tyrosine,
tryptophan, histidine, and others) can also absorb light and 3. Results 3.1. Light and Electron Microscopy of Age-Related Structural
Changes in the Rat Skin. The thickening of the epidermis,
with morphological alterations of the stratum basale, the dis-
appearance of interkeratinocytes and dermoepidermal junc-
tions, empty perinuclear spots in keratinocytes, and the
disappearance of the basal lamina were evident with advanc-
ing age (Figures 1 and 2). Besides, the dermal matrix was
more disorganized and avascular with the reduced number
of fibroblasts in old, as compared to mature (adult) and
young skin (Figures 1 and 2). The mitochondrial destruction
was evident in keratinocytes, fibroblasts, and mastocytes
(Figures 1 and 2). 2.4. Immunohistochemistry. Semifine (1 μm thick) sections
were used for proliferating cell nuclear antigen (PCNA)
and MsrA, HNE, and 8-oxoG detection by routine immu-
nohistochemistry. After removal of Araldite with 1% sodium
ethoxide in absolute ethanol (40 min, 37°C), skin sections
were rehydrated and subjected to heat-mediated epitope
retrieval and 3% hydrogen peroxide to block endogenous
peroxidase staining. Subsequently, slides were immersed in
protein block for 30 min at room temperature. After
thorough rinsing, sections were incubated overnight at 4°C
with anti-HNE (1 : 400; ab48506; Abcam, Cambridge, UK),
anti-MsrA
(1μgml−1;
ab16803,
Abcam),
anti-8-oxoG
(1 : 100; ab65548, Abcam), or anti-PCNA (1 : 100; sc7907;
Santa Cruz Biotechnology, Santa Cruz, CA, USA) antibodies. After rinsing in PBS, the sections were incubated with
the avidin-biotin-peroxidase complex (ABC-peroxidase kit,
Vector Labs, Burlingame, CA, USA), according to the manu-
facturer’s instructions. After rinsing in PBS, the peroxidase
reaction was developed using 3′-3′-diaminobenzidine tetra-
hydrochloride (Sigma-Aldrich, St. Louis, MO, USA) in the
presence of hydrogen peroxide for 7 min in the dark. Nega-
tive controls were performed for each set of experiments by
omitting primary antibodies and substituting them with
PBS. All sections were counterstained with haematoxylin,
dehydrated, and mounted for the analysis with an optical
light microscope (Leica DMLB microscope, Leica Microsys-
tems). The quantitative evaluation of antibody staining
intensity in skin sections was analyzed as described previ-
ously [28] using the IHC Profiler plugin and open-resource
digital image analysis software, ImageJ. 3.2. Ultrastructural
Characteristics
of
Fibroblasts
and
Collagen during Lifetime. Numerous elongated fibroblasts
fulfill the dermis of 15-day-old rats (Figure 2(a)). The
extracellular matrix is poorly organized, and sparse thin
collagen bundles are visualized between fine fibroblast
extensions (arrows in Figure 2(a)). The dermis in an adult
3-month-old rat consisted of large fibroblasts with well-
developed rough endoplasmic reticulum (ER). 2. Material and Methods 2.1. Animals and Sample Preparations. The experiments were
approved by the Ethical Committee for the Treatment of
Experimental Animals of the Institute for Biological Research Oxidative Medicine and Cellular Longevity 3 of the disulfide bond in the 5,5′-dithio-bis-nitrosobenzoic
acid (DTNB), with an increase in the absorbance at 412 nm
over the first two minutes. The activity of the enzyme is
determined by the amount of NADPH that is oxidized in a
minute, since 1 mol of NADPH produces 2 mol of thienitro-
benzoate and is expressed as a specific activity in nM
NADPH min−1 mg−1 protein. with 1% sodium ethoxide in absolute ethanol (40 min,
37°C), skin sections were rehydrated and stained with propi-
dium iodide for 10 min. After rinsing in distilled water, the
sections were mounted in Mowiol solution (Polysciences,
Eppelheim, Germany) in order to preserve fluorescence sig-
nal and examined with a Carl Zeiss LSM 510 confocal laser
scanning microscope (Carl Zeiss, Oberkochen, Germany). 2.3. Morphological Analysis by Light and Electron Microscopy. Immediately after isolation, skin samples were fixed in
2.5% glutaraldehyde in 0.1 M Sørensen phosphate buffer
(pH 7.2), postfixed in 2% osmium tetroxide in the same
buffer, routinely dehydrated using increasing concentrations
of ethanol, and embedded in Araldite (Sigma-Aldrich
Laborchemikalien GmbH, Hamburg, Germany) as done
previously [27]. Blocks were trimmed and cut into 1μm
sections using a Leica UC6 ultramicrotome (Leica Micro-
systems, Wetzlar, Germany), mounted on copper grids,
and contrasted in uranyl acetate and lead citrate using Leica
EM STAIN (Leica Microsystems). Sections were examined
on a Philips CM12 transmission electron microscope
(Philips/FEI, Eindhoven, Netherlands) equipped with a
digital camera (SIS MegaView III, Olympus Soft Imaging
Solutions, Münster, Germany). 2.6. Statistics. Statistical analyses were performed by one-way
ANOVA followed by Dunnett’s post hoc test using Prism
version 5.00 software (GraphPad Software, USA). Measure-
ments and quantifications are shown as mean ± SEM. Signif-
icance levels among different groups are indicated within the
figure legends. An asterisk (∗) is used for indicating signifi-
cance levels between an adult rat (3 m) and aging groups
(3d-21 m). The p value < 0.05 was regarded as significant. 3. Results Dilated ER
cisterns are filled with proteinaceous material; the extracellu-
lar matrix is well organized, showing compact and regularly
oriented collagen bundles (Figure 2(b)). However, in the
dermis of a 21-month-old rat, fibroblasts reduced in number
and their long cytoplasmic extensions wrap large collagen
bundles with thicker and tightly packed, but poorly oriented
and electron-lucent, fibers (Figure 2(c)). 3.3. Proliferative Capacity of Skin Cells during Lifetime. Immunohistochemical analysis revealed the highest expres-
sion of PCNA in the skin of postnatal 3-day-old rats
(Figure 3). At this age, a plethora of PCNA-immunopositive
nuclei were evident in the epidermis and dermis (Figure 3(a),
inset). The expression of PCNA was detected in nuclei of
basal keratinocytes, scattered dermal fibroblasts, and cells
of the dermal papilla in adult (3-month-old) rats. The PCNA
immunopositivity was evidently diminished in the epidermis
and dermis of elderly (21-month-old) rat skin (p < 0 01). 2.5. Propidium Iodide Staining. Semithin (1 μm thick) sec-
tions were used for the analysis of chromatin condensation
and apoptotic changes of nuclei. After removal of Araldite 4 Oxidative Medicine and Cellular Longevity 3 d
15 d
1 m
3 m
6 m
12 m
21 m
Figure 1: Light and electron microscopy of age-related structural changes in the rat skin. In the young skin, all epidermal cell layers have a
normal structure and all cells appeared to be healthy (3 days, 15 days, 1 month, and 3 months old). Aging (6-month-, 12-month-, and
21-month-old skin) increases the epidermal layer thickness and the intercellular space throughout the epidermis along with a decrease in
epithelial-dermal junction. In old skin, the basal lamina was partially detached and disrupted from the basal cell layer. Desquamation,
through reduction in intercellular lipid contents, was seen in the upper layer of the stratum corneum. Intracytoplasmic vacuolization in
the keratinocytes increased with aging. Namely, the keratinocyte mitochondria become light and swollen and concentrated around the
nucleus to form an electron-lucent structure—“nuclear halo” (black arrow, 21 m, right insets). The numerous small cytoplasmic vacuoles,
located near the nucleus, were observed in the basal keratinocytes gradually becoming a large cytoplasmic vacuole. Degenerative cells with
a dark cytoplasm (white arrow, 21 m) were also seen in the basal layer of the epidermis. Semifine sections stained with toluidine blue,
magnification: ×100, orig.; transmission electron microscopy, magnification: ×8000, orig. 3. Results 3 d
15 d
1 m
3 m 6 m Figure 1: Light and electron microscopy of age-related structural changes in the rat skin. In the young skin, all epidermal cell layers have a
normal structure and all cells appeared to be healthy (3 days, 15 days, 1 month, and 3 months old). Aging (6-month-, 12-month-, and
21-month-old skin) increases the epidermal layer thickness and the intercellular space throughout the epidermis along with a decrease in
epithelial-dermal junction. In old skin, the basal lamina was partially detached and disrupted from the basal cell layer. Desquamation,
through reduction in intercellular lipid contents, was seen in the upper layer of the stratum corneum. Intracytoplasmic vacuolization in
the keratinocytes increased with aging. Namely, the keratinocyte mitochondria become light and swollen and concentrated around the
nucleus to form an electron-lucent structure—“nuclear halo” (black arrow, 21 m, right insets). The numerous small cytoplasmic vacuoles,
located near the nucleus, were observed in the basal keratinocytes gradually becoming a large cytoplasmic vacuole. Degenerative cells with
a dark cytoplasm (white arrow, 21 m) were also seen in the basal layer of the epidermis. Semifine sections stained with toluidine blue,
magnification: ×100, orig.; transmission electron microscopy, magnification: ×8000, orig. 3.4. Time-Course Changes in the Level of HNE-Modified
Proteins in the Rat Skin. The highest level of HNE-protein
adducts
was
observed
in
postnatal
3-day-old
rats
(Figure 4(b)). This level was higher compared to that in
young adult 3-month-old rat (p < 0 001) and all other ages
(Figures 4(a) and 4(b)). In 3-day-old skin, strong HNE
immunopositivity was observed both in the cytoplasm and
in the nuclei of epidermal and dermal cells (Figure 4(a)). In adult (3-month-old) rat skin, only nuclei in the stratum
basale were faintly HNE-positive. In comparison to young
adult skin, an increased granular, mostly cytoplasmic
reaction for HNE was evident in 21-month-old rat skin
(p < 0 05). Intensive HNE immunoreactivity in the basal
lamina
was
also
apparent
in
some
areas
of
the
21-month-old skin (Figure 4(c), right inset). 3.4. Time-Course Changes in the Level of HNE-Modified
Proteins in the Rat Skin. The highest level of HNE-protein
adducts
was
observed
in
postnatal
3-day-old
rats
(Figure 4(b)). This level was higher compared to that in
young adult 3-month-old rat (p < 0 001) and all other ages
(Figures 4(a) and 4(b)). 3. Results Dilated ER cisterns are filled with proteinaceous material; the extracellular matrix is well organized,
showing compact and regularly oriented collagen bundles (b). In the dermis of 21-month-old rat, the fibroblasts reduced in number and
their long cytoplasmic extensions wrap large collagen bundles with thicker and tightly packed, but poorly oriented and electron-lucent,
fibers (c). Magnification: semifine section, toluidine blue staining: ×100, orig.; framed area analyzed on transmission electron microscopy,
magnification: ×3000, orig.; far right: ×8800, orig. fb fb
fb
(a) (a) fb fb
(b) (b) fb fb
(c) (c) Figure 2: Lifetime ultrastructural characteristics of fibroblasts and collagen. The dermis in the skin of 15-day-old rats is occupied with
numerous elongated fibroblasts (a). The extracellular matrix is poorly organized, and sparse thin collagen bundles are visualized between
fine fibroblast extensions (arrows). The mature skin dermis in adult 3-month-old rat (b) consisted of large fibroblasts with well-developed
rough endoplasmic reticulum (ER). Dilated ER cisterns are filled with proteinaceous material; the extracellular matrix is well organized,
showing compact and regularly oriented collagen bundles (b). In the dermis of 21-month-old rat, the fibroblasts reduced in number and
their long cytoplasmic extensions wrap large collagen bundles with thicker and tightly packed, but poorly oriented and electron-lucent,
fibers (c). Magnification: semifine section, toluidine blue staining: ×100, orig.; framed area analyzed on transmission electron microscopy,
magnification: ×3000, orig.; far right: ×8800, orig. activity of GSH-Px was higher in the oldest groups of rats, as
compared to the young adult group (3 months old). increases in CAT, GSH-Px, and GR are associated with a
decline in protein expression of MsrA, accumulation of
HNE-modified protein aldehydes, particularly in the basal
lamina, and 8-oxoG positivity in nuclei and mitochondria. Such redox profile in old skin is accompanied by degenera-
tive changes at structural and ultrastructural levels in epider-
mal and dermal compartments (thickening of the epidermis,
occurrence of inter- and intracellular empty spaces through-
out the epidermis, disappearance of the basal lamina, loss of
dermoepidermal junction, rarefaction of fibroblasts, and
enlargement and thickening of collagen bundles) and low
renewal skin capacity (according to a decrease in the number
of PCNA-positive cells). In 3-day-old skin, higher SOD,
CAT, GR, and TR activities and MsrA protein expression
correspond to intensive postnatal development and prolifer-
ation. 3. Results In 3-day-old skin, strong HNE
immunopositivity was observed both in the cytoplasm and
in the nuclei of epidermal and dermal cells (Figure 4(a)). In adult (3-month-old) rat skin, only nuclei in the stratum
basale were faintly HNE-positive. In comparison to young
adult skin, an increased granular, mostly cytoplasmic
reaction for HNE was evident in 21-month-old rat skin
(p < 0 05). Intensive HNE immunoreactivity in the basal
lamina
was
also
apparent
in
some
areas
of
the
21-month-old skin (Figure 4(c), right inset). including the stratum basale (Figure 5(a)). In comparison
to young adult skin, immunopositivity of 8-OxoG was
higher in 21-month-old rat skin (p < 0 01) (Figure 5(b)). Besides, the mitochondrial positivity of 8-OxoG is observed
in 21-month-old skin, and this positivity corresponds to
apoptotic positivity observed by propidium iodide staining
in the sebaceous glands and root sheath (Figure 5(c)). 3.6. Age-Dependent Changes of the SOD, CAT, GSH-Px, GR,
and TR Activities in the Skin of Rats. In comparison to adult
(3-month-old) skin, total SOD and TR activities were higher
in the preadult period: SOD activity was higher in 3-day- and
1-month-old groups (p < 0 001 and p < 0 01), while TR was
significantly higher in 3-day-, 15-day- and 1-month-old
groups (p < 0 001) (Figure 6). Thereafter, activities of SOD
and TR remain constant through adulthood and old age. Activities of CAT and GR decrease sharply towards adult-
hood (2-3 m) and mostly remain constant in middle-aged
rat skin but thereafter increase again in the oldest groups
(18-month- and 21-month-old groups). In a similar fashion, 3.5. Analysis of the DNA Lesion and 8-OxoG in the Skin of
Rats during the Lifetime. Aging increases the number of
8-OxoG-immunopositive nuclei in epidermal cells (from
1 m to 21 m), but only in the skin of 21-month-old rats is
immunopositivity of 8-OxoG present in all epidermal layers, 5 Oxidative Medicine and Cellular Longevity 5 fb
fb
fb
(a)
fb
fb
(b)
fb
fb
(c)
Figure 2: Lifetime ultrastructural characteristics of fibroblasts and collagen. The dermis in the skin of 15-day-old rats is occupied with
numerous elongated fibroblasts (a). The extracellular matrix is poorly organized, and sparse thin collagen bundles are visualized between
fine fibroblast extensions (arrows). The mature skin dermis in adult 3-month-old rat (b) consisted of large fibroblasts with well-developed
rough endoplasmic reticulum (ER). 3. Results Moreover, the high level of HNE characterizes young
skin, but it is not accompanied by any signs of oxidative
insults on DNA, cell, or tissue structure levels. The overall
results of this study indicate that “positive” oxidative pres-
sure (or eustress) seems to be integral in adaptive physiology
of a newborn’s skin. Also, progressively compromised redox 3.7. Immunoexpression of MsrA. The highest level of MsrA is
detected in 3-day-old (early postnatal) skin (Figure 7(a)). Strong immunostaining for MsrA is evident in both epider-
mal and dermal skin compartments. The MsrA expression
decreases with passage of life, and the lowest level of MsrA
is observed in the skin of old 21-month-old rats (Figure 7(b)). 4. Discussion The relative mean intensity was determined from 6 images for each aging group (n = 6), and it is
shown graphically (b). Data were presented as mean ± SEM. ∗In respect to 3-month-old rats; ∗p < 0 05; ∗∗p < 0 01. ×100 magnification,
orig.; corner insets show low magnification (×40, orig.) of representative skin sections. of molecular oxidative damage (oxidative modifications of
membrane lipids, DNA, and proteins) accelerates replica-
tive senescence of skin fibroblasts in vitro and aging
in vivo [37–39]. In turn, senescent fibroblasts are a source
of ROS, as reviewed in [40]. Thus, a vicious cycle between
ROS-induced senescence and increased ROS due to accumu-
lation of senescent cells could be established in aged skin. flexibility and/or repair, rather than an overall drop in ROS
scavenging capacity and/or severe oxidative injuries, reflect
acceleration of chronological aging of the skin in adults. The details of the redox states in the skin and their correla-
tions with morphofunctional changes in the function of life-
time are discussed in more detail below. Histology of old animals shows typical aging changes in
the structural organization of the skin, and all of them can
result from the action of free radicals. Damage to the extra-
cellular matrix by free radicals, especially hyaluronic acid
and proteoglycans [30], can result in the thickening of the
epidermis and the empty spaces between keratinocytes, as
noted in old skin (Figure 1). Within the keratinocytes, mito-
chondria become light, swollen, and concentrated around the
nucleus resulting in the occurrence of degenerative changes,
known as perinuclear “halo” formations (Figure 1). This his-
tological phenomenon also occurs in UV-irradiated skin, and
it closely correlates with ROS levels, since pretreatment with
SOD restricts the formation of “halo” spaces [31]. The loss of
normal maturation and irregularities in the epidermal layers,
as noted here in the skin of old rats, is common in photo-
damaged skin and is also connected to free radical action
[15, 32]. In the dermis of old rats, a decline in collagen
fiber content and density is pronounced (Figure 2). It is well
known that increased ROS, as in inflammatory conditions,
can affect collagen density directly by damaging fibers and
indirectly by interfering with collagen synthesis [33] or fibro-
blast proliferation and increasing senescence [34, 35]. In parallel with morphofunctional manifestations of det-
rimental ROS action, we examined the level of HNE-protein
adducts. 4. Discussion Aging studies were mostly based on comparison of AD and
lipid peroxidation in only two age groups of animals [19]
or fibroblasts from young and old donors [29]. In this study,
the relation of redox and structural alterations in skin aging
was examined in light of chronological aging, in discrete time
intervals since birth (3rd day) to 21 months of rat age. In
comparison to adult (3-month-old) skin, higher activities of
enzymes involved in ROS removal are observed in young
(3-day-old) and old (21-month-old) skin. In old skin, Oxidative Medicine and Cellular Longevity 6 3 d
3 m
15 d
1 m
6 m
21 m
3 d
3m
21 m
20 휇m
50 휇m
(a)
Proliferating cell nuclear antigen
0
3 d
15 d
1 m
3 m
6 m 21 m
60
120
Lifetime
PCNA staining quantification
(%)
⁎
⁎⁎
(b)
Figure 3: Expression of proliferating cell nuclear antigen (PCNA) in the skin of 3-day-, 15-day-, 1-month-, 3-month-, 6-month-, and
21-month-old rats. Tissues were subjected to immunohistochemistry using a PCNA-specific antibody. The brown nuclei testify to the
specific staining (a). The quantitative evaluation of PCNA antibody staining intensity in skin sections was analyzed by the IHC Profiler
using the open-source ImageJ program. The relative mean intensity was determined from 6 images for each aging group (n = 6), and it is
shown graphically (b). Data were presented as mean ± SEM. ∗In respect to 3-month-old rats; ∗p < 0 05; ∗∗p < 0 01. ×100 magnification,
orig.; corner insets show low magnification (×40, orig.) of representative skin sections. 15 d
6 m휇
(a) 3 d
3 d
20 휇m
50 휇m 1 m
21 m
21 m Proliferating cell nuclear antigen
0
3 d
15 d
1 m
3 m
6 m 21 m
60
120
Lifetime
g qi
(%)
⁎
⁎⁎
(b) Proliferating cell nuclear antigen PCNA staining quantification
( ) 3 m
3m Lifetime (b) Figure 3: Expression of proliferating cell nuclear antigen (PCNA) in the skin of 3-day-, 15-day-, 1-month-, 3-month-, 6-month-, and
21-month-old rats. Tissues were subjected to immunohistochemistry using a PCNA-specific antibody. The brown nuclei testify to the
specific staining (a). The quantitative evaluation of PCNA antibody staining intensity in skin sections was analyzed by the IHC Profiler
using the open-source ImageJ program. 4. Discussion The immunohistochemical method was used to detect skin tissue proteins modified by 4-hydroxynonenal (HNE)
ubsequent quantitative image analysis (b). The brown staining testifies to the specific reaction. Strong HNE immunopositivity of the ba
amina, along with its partial discontinuation/disruption (black arrows), was observed in 21-month-old rat skin (c, left inset, arrow
Dermoepidermal junctions and the basal cell deep projections into the dermis were also disturbed (c, middle and right, arrows). Representative slides of HNE immunohistochemistry are shown at ×40 (corner inset) and ×100 magnification, orig. The quantitat
valuation of HNE antibody staining intensity in skin sections (b) was analyzed as described in Figure 3. (c) Magnification: semifi
ection, toluidine blue staining: ×100, orig. Insets: left—HNE immunohistochemistry: ×100 orig.; right—transmission electron microsco
bar: 2 μm. The relative mean intensity was determined from 6 images for each aging group (n = 6). Data were presented as mean ± SE
In respect to 3-month-old rats; ∗p < 0 05; ∗∗∗p < 0 001. Oxidative Medicine and Cellular Longevity 3 d
15 d
1 m
3 m
6 m
21 m
3 d
20 휇m
50 휇m
(a) 21 m (a) (a)
6 m
21 m
⁎
2 휇m
(c) ( )
3 d
15 d
1 m
3 m
6 m
21 m
20
40
60
80
Lifetime
4-HNE staining quantifivation
(%)
4-Hydroxynonenal
⁎⁎⁎
⁎
(b)
2 휇m
(c) 3 d
15 d
1 m
3 m
6 m
21 m
20
40
60
80
Lifetime
4-HNE staining quantifivation
(%)
4-Hydroxynonenal
⁎⁎⁎
⁎
(b) (c) (b) Figure 4: Time-course changes in the levels of HNE-modified proteins in the skin of 3-day-, 15-day-, 1-month-, 3-month-, 6-month-, and
21-month-old rats. The immunohistochemical method was used to detect skin tissue proteins modified by 4-hydroxynonenal (HNE) for
subsequent quantitative image analysis (b). The brown staining testifies to the specific reaction. Strong HNE immunopositivity of the basal
lamina, along with its partial discontinuation/disruption (black arrows), was observed in 21-month-old rat skin (c, left inset, arrows). Dermoepidermal junctions and the basal cell deep projections into the dermis were also disturbed (c, middle and right, arrows). (a)
Representative slides of HNE immunohistochemistry are shown at ×40 (corner inset) and ×100 magnification, orig. The quantitative
evaluation of HNE antibody staining intensity in skin sections (b) was analyzed as described in Figure 3. (c) Magnification: semifine
section, toluidine blue staining: ×100, orig. 4. Discussion HNE, produced during the lipid peroxidation pro-
cess, covalently binds to proteins to form Michael adducts
[41]. An increase in HNE-modified proteins is common
upon serial passaging of human keratinocytes [42] or replica-
tive senescence of WI-38 fibroblasts [35] and in the retina of
aged rats [43]. Thus, HNE is an important marker and prop-
agator of lipid, DNA, and protein oxidation [44–46] and is of
particular importance in aging skin. Results of this study
show that in comparison to adult skin, HNE immunopositiv-
ity increases in old age (after 21 months). The observed
increase in HNE immunopositivity in old skin could however
be “diluted,” since skin consists of many layers and struc-
tures, and modifications of some of them may severely inflict
the whole skin architecture. In line with this, strong HNE
immunopositivity detected on the basal lamina, uneven in
some parts of skin, may contribute to its deterioration with
advanced aging (as shown in Figure 4(c)). An increase in HNE-modified proteins with aging could
be regarded as a consequence of diminished antioxidant pro-
tection. However, no significant changes were found in the
case of SOD and TR, and the activities of CAT, GSH-Px,
and GR even show an increase in old (18 m-21 m) rat skin
(Figure 6). Although studies reported species-, strain-, sex-,
and tissue-specific age-related changes in AD [47], the skin The senescent phenotype is characterized by low
expression of proteins required for DNA synthesis and
low proliferation rate [36]. In agreement, the lower number
of PCNA-positive nuclei in the dermis and epidermis is
found in 21-month-old rat skin (Figure 3). Accumulation 7 Oxidative Medicine and Cellular Longevity of old mice [19] and fibroblasts from old human donors [29]
tl
h
t i
d b
h
d
i
d AD
imperfect ROS scavenging or rather it is predominan
d t
i
d b i
i d d
d ti
f HNE
difid
3 d
15 d
1 m
3 m
6 m
21 m
3 d
20 휇m
50 휇m
(a)
3 d
15 d
1 m
3 m
6 m
21 m
20
40
60
80
Lifetime
4-HNE staining quantifivation
(%)
4-Hydroxynonenal
⁎⁎⁎
⁎
(b)
2 휇m
(c)
Figure 4: Time-course changes in the levels of HNE-modified proteins in the skin of 3-day-, 15-day-, 1-month-, 3-month-, 6-month-, a
21-month-old rats. 4. Discussion 20 휇m
3 d
15 d
1 m
3 m
6 m
21 m
8-Oxoguanine
0
3 d
15 d
1 m
3 m
6 m
21 m
20
40
60
Lifetime
8-OxoG staining quantification
(%)
⁎⁎
⁎⁎
⁎⁎⁎ 20 휇m
3 d
15 d
1 m
3 m
6 m
21 m 8-Oxoguanine
0
3 d
15 d
1 m
3 m
6 m
21 m
20
40
60
Lifetime
8-OxoG staining quantification
(%)
⁎⁎
⁎⁎
⁎⁎⁎ Lifetime (a)
3 m (a)
(b)
3 m
21 m
(c) (a) (b) (c) Figure 5: Analysis of the DNA lesion and 8-oxoguanine (8-OxoG) in the skin of rats during the lifetime. Tissues were subjected to
immunohistochemistry using an 8-OxoG-specific antibody. The brown staining testifies to the specific reaction (a). The quantitative
evaluation of 8-OxoG antibody staining intensity in skin sections was analyzed as described in Figure 3 (b). The relative mean intensity
was determined from 6 images for each aging group (n = 6). Data were presented as mean ± SEM. ∗In respect to 3-month-old rats;
∗∗p < 0 01; ∗∗∗p < 0 001. The mitochondrial positivity of 8-OxoG corresponds to apoptotic positivity observed by propidium iodide
staining in the sebaceous glands and root sheath (c, right). Magnification: ×100, orig. correlation with morphofunctional characteristics of the skin
in discrete time intervals during life, including the changes
immediately after birth. Trx-dependent reduction of methionine sulfoxide to methio-
nine [50, 51]. Thus, a decline in MsrA expression in old rats,
as observed here in the skin of old rats (Figure 7) and other
aging tissues or senescent cells [52, 53], suggests that a
decline in protein repair contributes to the accumulation of
oxidized proteins and impaired redox sensitivity/regulation
with chronological skin aging, regardless of overall upregula-
tion of AD enzyme activities. Moreover, a progressive, gener-
alized impairment of oxidative repair could result in an
increase in 8-oxoG positivity in nuclei of basal keratinocytes
and mitochondria in the secretory cells of sebaceous glands
observed in the 21-month-old skin (Figure 5). According to a plethora of PCNA-positive nuclei, the
earliest postnatal development is characterized by the
intensive proliferation rate of skin cells. Intensive prolifer-
ation enables thickening of the outer layer (stratum cor-
neum) and dermis (growth of hair follicles, sebaceous
and sweat glands) as presented in Figures 1–3. 4. Discussion Insets: left—HNE immunohistochemistry: ×100 orig.; right—transmission electron microscopy,
bar: 2 μm. The relative mean intensity was determined from 6 images for each aging group (n = 6). Data were presented as mean ± SEM. ∗In respect to 3-month-old rats; ∗p < 0 05; ∗∗∗p < 0 001. of old mice [19] and fibroblasts from old human donors [29]
are mostly characterized by unchanged or increased AD
enzyme activities. Moreover, fibroblasts from old donors
are more resistant to oxidative pressure from hydrogen per-
oxide, linoleic acid hydroperoxide, or UV light, presumably
because of higher activity of GSH-Px in these cells [29]. It
remains vague however whether the increase in HNE,
observed in the current study in old skin, is associated with imperfect ROS scavenging or rather it is predominantly
determined by impaired degradation of HNE-modified pro-
teins in aging skin, as suggested [42], or by both. In addition,
concomitant with accumulation of oxidative damages on
molecular
and
supramolecular
levels,
the
skin
from
21-month-old rats is characterized by low expression of
MsrA. This enzyme is essential in the maintenance of func-
tional proteins in aerobic cells [48, 49] since it catalyzes Oxidative Medicine and Cellular Longevity 8 8 20 휇m
3 d
15 d
1 m
3 m
6 m
21 m
(a)
8-Oxoguanine
0
3 d
15 d
1 m
3 m
6 m
21 m
20
40
60
Lifetime
8-OxoG staining quantification
(%)
⁎⁎
⁎⁎
⁎⁎⁎
(b)
3 m
21 m
(c)
re 5: Analysis of the DNA lesion and 8-oxoguanine (8-OxoG) in the skin of rats during the lifetime. Tissues were subjected to
unohistochemistry using an 8-OxoG-specific antibody. The brown staining testifies to the specific reaction (a). The quantitative
ation of 8-OxoG antibody staining intensity in skin sections was analyzed as described in Figure 3 (b). The relative mean intensity
determined from 6 images for each aging group (n = 6). Data were presented as mean ± SEM. ∗In respect to 3-month-old rats;
0 01; ∗∗∗p < 0 001. The mitochondrial positivity of 8-OxoG corresponds to apoptotic positivity observed by propidium iodide
ng in the sebaceous glands and root sheath (c, right). Magnification: ×100, orig. 4. Discussion U mg protein−1
0
20
40
60
CAT
f
3 d
15 d
1 m
2 m
3 m
6 m
9 m
12 m
15 m
18 m
21 m
⁎⁎⁎
⁎
⁎⁎⁎ ⁎ 0
20
40
60
80
nM NADPH min−1 mg protein−1
GSH-Px
Lifetime
3 d
15 d
1 m
2 m
3 m
6 m
9 m
12 m
15 m
18 m
21 m
⁎ ⁎⁎ nM NADPH min−1 mg protein−1 ⁎⁎ U mg protein−1 nM NADPH min 1 mg protein 1
0
2
4
6
8
10
TR
Lifetime
3 d
15 d
1 m
2 m
3 m
6 m
9 m
12 m
15 m
18 m
21 m
Lifetime
1
1
1
2
⁎⁎⁎
⁎⁎⁎
⁎⁎⁎ Lifetime Lifetime
0
20
40
60
80
nM NADPH min−1 mg protein−1
GR
Lifetime
3 d
15 d
1 m
2 m
3 m
6 m
9 m
12 m
15 m
18 m
21 m
⁎⁎
⁎
⁎ nM NADPH min−1 mg protein−1 Lifetime Lifetime Lifetime Figure 6: Age-dependent changes of the total superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione
reductase (GR), and thioredoxin reductase (TR) activities in the skin of rats. Data were presented as mean ± SEM. ∗In respect to
3-month-old rats; ∗p < 0 05; ∗∗p < 0 01; ∗∗∗p < 0 001. Figure 6: Age-dependent changes of the total superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione
reductase (GR), and thioredoxin reductase (TR) activities in the skin of rats. Data were presented as mean ± SEM. ∗In respect to
3-month-old rats; ∗p < 0 05; ∗∗p < 0 01; ∗∗∗p < 0 001. 15 d휇 15 d
1 m
3 d
3 m
6 m
21 m
50 휇m
(a)
Methionine sulfoxide reductase A
0
3 d
15 d 1 m 3 m 6 m 21 m
20
40
60
80
Lifetime
MsrA staining quantification
(%)
⁎
⁎
(b)
Figure 7: Expression of methionine sulfoxide reductase A (MsrA) in the skin of 3-day-, 15-day-, 1-month-, 3-month-, 6-month-, and
21-month-old rats. Tissues were subjected to immunohistochemistry using an MsrA-specific antibody. The brown staining testifies to the
specific reaction. The quantitative evaluation of MsrA antibody staining intensity in skin sections (b) was analyzed as described in
Figure 3. The relative mean intensity was determined from 6 images for each aging group (n = 6). 4. Discussion These pro-
cesses are essential in the development of skin barrier
function—defense of the organism against low/high tem-
perature, oxygen, and UV radiation after birth. It seems
that this adaptive period in rats lasts for a few weeks, since
PCNA immunopositivity progressively diminished starting
from 15 days on (Figure 3). A similar time frame exists in To better understand the process of chronological skin
aging, the current time-course study puts forward the
dynamics of changes in the skin redox state and its Oxidative Medicine and Cellular Longevity 9 ⁎⁎⁎
0
10
20
30
40
Lifetime
U mg protein−1
U mg protein−1
0
20
40
60
0
20
40
60
80
0
20
40
60
80
nM NADPH min−1 mg protein−1
nM NADPH min−1 mg protein−1
nM NADPH min−1 mg protein−1
0
2
4
6
8
10
SOD
CAT
GSH-Px
GR
TR
3 d
15 d
1 m
2 m
3 m
6 m
9 m
12 m
15 m
18 m
21 m
Lifetime
3 d
15 d
1 m
2 m
3 m
6 m
9 m
12 m
15 m
18 m
21 m
Lifetime
3 d
15 d
1 m
2 m
3 m
6 m
9 m
12 m
15 m
18 m
21 m
Lifetime
3 d
15 d
1 m
2 m
3 m
6 m
9 m
12 m
15 m
18 m
21 m
Lifetime
3 d
15 d
1 m
2 m
3 m
6 m
9 m
12 m
15 m
18 m
21 m
⁎⁎⁎
⁎⁎
⁎⁎⁎
⁎
⁎⁎⁎ ⁎
⁎ ⁎⁎
⁎⁎⁎
⁎⁎⁎
⁎⁎⁎
⁎⁎
⁎
⁎
Figure 6: Age-dependent changes of the total superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione
reductase (GR), and thioredoxin reductase (TR) activities in the skin of rats. Data were presented as mean ± SEM. ∗In respect to
3-month-old rats; ∗p < 0 05; ∗∗p < 0 01; ∗∗∗p < 0 001. Oxidative Medicine and Cellular Longevity Oxidative Medicine and Cellular Longevity References [1] D. Harman, “Aging: a theory based on free radical and radia-
tion chemistry,” Journal of Gerontology, vol. 11, no. 3,
pp. 298–300, 1956. [2] H. Yoshimura, “Organ system in adaptation: the skin,” in
Handbook of Physiology, C. G. Wilber, Ed., pp. 109–133,
American Physiological Society, Washington, D.C., 1963. p
p y
g
g
g
p
In summary, the results of this study indicate that chro-
nological aging of the skin is associated with slowly advancing
redox “inflexibility” and repair, rather than with an overall
drop in ROS-scavenging capacity. Although similar internal
and external sources may increase ROS in young and senes-
cent skin, the mechanisms for their removal are different,
and so is the result accordingly: proliferation/regeneration
in postnatal skin and senescence and atrophy in the elderly. Understanding the regulation of redox-recycling-redox sys-
tems, like those with reductase and denitrosilase activity
(Msr, GSH/GR, and thioredoxin/thioredoxin reductase) and
genome-repairing systems (such as 8-oxoguanine DNA gly-
cosylase), can advance healthy aging of the skin. [3] G. Nikolakis, E. Makrantonaki, and C. C. Zouboulis, “Skin
mirrors human aging,” Hormone Molecular Biology and Clin-
ical Investigation, vol. 16, no. 1, pp. 13–28, 2013. [4] A. M. Michelson, “Liposomal SOD and its application to skin
diseases and other disorders,” Japanese Journal of Dermatol-
ogy, vol. 96, pp. 1376–1383, 1986. [5] D. Darr and I. Fridovich, “Free radicals in cutaneous biol-
ogy,” Journal of Investigative Dermatology, vol. 102, no. 5,
pp. 671–675, 1994. [6] A. M. Pathak, “Photosensitivity to drugs,” in Drugs and
Enzymes, B. B. Brodie and J. R. Gillette, Eds., vol. 3, pp. 419–
439, Pergamon Press, Czechoslovak Medical Press, 1963. [7] B. A. Jurkiewicz and G. R. Buettner, “Ultraviolet light-induced
free radical formation in skin: an electron paramagnetic reso-
nance study,” Photochemistry and Photobiology, vol. 59,
no. 1, pp. 1–4, 1994. Data Availability mice where the maximal proliferation of dermal fibroblasts
is seen in newborns, but it decreased as early as the third
postnatal week, the same time when the dermis reaches its
maximal thickness [54]. The data used to support the findings of this study are
included within the article. [
]
In parallel, marked HNE immunopositivity in early post-
natal skin (3 days old), as seen in Figure 4, could reflect the
intensive metabolic activity needed for ongoing growth, pro-
liferation, and morphogenesis of epidermal layers and der-
mal structures (like hair follicles and glands). Also, higher
HNE may be connected to the increase in ROS production
imposed by exposure of the skin to a new environment, oxy-
gen, and UV light, following the transition from intrauterine
to extrauterine environment. These external factors seem to
be important sources of lipid peroxides and HNE in skin
[55]. A “physiologically relevant increase” in HNE levels,
triggered by a rise in partial oxygen pressure, could support
proliferation/cell turnover in postnatal skin. Accordingly,
higher levels of HNE and AD in postnatal skin, as compared
to adult skin, are not surprising. The role of HNE in prolifer-
ation and differentiation of cells in vitro is well recognized
but complex, which is dependent on multitude variables,
including the cellular level of HNE, redox environment, and
cell type [56–58]. Thus, additional studies are needed to com-
prehend physiological signaling of HNE in postnatal skin. Conflicts of Interest The authors declare that there is no conflict of interest
regarding the publication of this article. Acknowledgments This work was supported by the Ministry of Education,
Science and Technological Development of the Republic
of Serbia (Grant No. 173055). 4. Discussion Data were presented as mean ± SEM ∗
In respect to 3-month-old rats; ∗p < 0 05. Representative slides are shown at ×40 magnification, orig. 15 d
1 m
3 d
3 m
6 m
21 m
50 휇m
Methionine sulfoxide reductase
0
3 d
15 d 1 m 3 m 6 m 21
20
40
60
80
Lifetime
MsrA staining quantification
(%)
⁎
⁎ 21 m Methionine sulfoxide reductase A
0
3 d
15 d 1 m 3 m 6 m 21 m
20
40
60
80
Lifetime
MsrA staining quantification
(%)
⁎
⁎
(b) 6 m
(a) Methionine sulfoxide reductase A 6 m Methionine sulfoxide reductase A 3 m (b) (a) Figure 7: Expression of methionine sulfoxide reductase A (MsrA) in the skin of 3-day-, 15-day-, 1-month-, 3-month-, 6-month-, and
21-month-old rats. Tissues were subjected to immunohistochemistry using an MsrA-specific antibody. The brown staining testifies to the
specific reaction. The quantitative evaluation of MsrA antibody staining intensity in skin sections (b) was analyzed as described in
Figure 3. The relative mean intensity was determined from 6 images for each aging group (n = 6). Data were presented as mean ± SEM ∗
In respect to 3-month-old rats; ∗p < 0 05. Representative slides are shown at ×40 magnification, orig. 10 5. Conclusion and Perspectives Gilchrest, “Fifty years of skin
aging,” Journal of Investigative Dermatology Symposium Pro-
ceedings, vol. 7, no. 1, pp. 51–58, 2002. [20] A. Jankovic, A. Korac, B. Srdic-Galic et al., “Differences in the
redox status of human visceral and subcutaneous adipose tis-
sues–relationships to obesity and metabolic risk,” Metabolism,
vol. 63, no. 5, pp. 661–671, 2014. [37] Q. Chen, A. Fischer, J. D. Reagan, L. J. Yan, and B. N. Ames,
“Oxidative DNA damage and senescence of human diploid
fibroblast cells,” Proceedings of the National Academy of Sci-
ences of the United States of America, vol. 92, no. 10,
pp. 4337–4341, 1995. [21] O. H. Lowry, N. J. Rosebrough, A. L. Farr, and R. J. Randall,
“Protein measurement with the Folin phenol
reagent,” Journal of Biological Chemistry, vol. 193, no. 1,
pp. 265–275, 1951. [38] P. Dumont, M. Burton, Q. M. Chen et al., “Induction of repli-
cative senescence biomarkers by sublethal oxidative stresses in
normal human fibroblast,” Free Radical Biology and Medicine,
vol. 28, no. 3, pp. 361–373, 2000. [22] H. P. Misra and I. Fridovich, “The role of superoxide anion in
the autoxidation of epinephrine and a simple assay for super-
oxide dismutase,” Journal of Biological Chemistry, vol. 247,
no. 10, pp. 3170–3175, 1972. [39] A. Terman, “Garbage catastrophe theory of aging: imperfect
removal of oxidative damage?,” Redox Report, vol. 6, no. 1,
pp. 15–26, 2001. [23] E. Beutler, “Red cell metabolism: a manual of biochemical
methods,” in Catalase, E. Beutler, Ed., pp. 105-106, Grune
and Stratton Inc, New York, 1982. [40] M. Kumar, W. Seeger, and R. Voswinckel, “Senescen-
ce-associated secretory phenotype and its possible role
in chronic obstructive pulmonary disease,” American Journal
of Respiratory Cell and Molecular Biology, vol. 51, no. 3,
pp. 323–333, 2014. [24] D. E. Paglia and W. N. Valentine, “Studies on the quantitative
and qualitative characterization erythrocyte glutathione per-
oxidases,” Journal of Laboratory and Clinical Medicine,
vol. 70, pp. 74–77, 1967. [41] K. Uchida, “4-Hydroxy-2-nonenal: a product and mediator of
oxidative stress,” Progress in Lipid Research, vol. 42, no. 4,
pp. 318–343, 2003. [25] D. Glatzle, J. P. Vuilleumier, F. Weber, and K. Decker, “Gluta-
thione reductase test with whole blood, a convenient proce-
dure for the assessment of the riboflavin status in humans,”
Experientia, vol. 30, no. 6, pp. 665–667, 1974. [42] I. Petropoulos, M. Conconi, X. 5. Conclusion and Perspectives Emerit and B. Chance, Eds., pp. 328–341,
Birkhauser Verlag, Basel, Switzerland, 1992. [14] L. C. Harber and R. L. Baer, “Pathogenic mechanisms of
drug-induced photosensitivity,” Journal of Investigative Der-
matology, vol. 58, no. 6, pp. 327–342, 1972. [31] I. Emerit, A. M. Michelson, E. Martin, and J. Emerit, “Perinuc-
lear halo formation as an indication of phototoxic effects,”
Dermatologica, vol. 163, no. 4, pp. 295–299, 1981. [15] K. Danno, T. Horio, M. Takigawa, and S. Imamura, “Role of
oxygen intermediates in UV-induced epidermal cell injury,”
Journal of Investigative Dermatology, vol. 83, no. 3, pp. 166–
168, 1984. [32] R. Rosario, G. J. Mark, J. A. Parrish, and M. C. Mihm Jr, “His-
tological changes produced in skin by equally erythemogenic
doses of UVA, UVB, UVC and UVA with psoralens,” British
Journal of Dermatology, vol. 101, no. 3, pp. 299–308, 1979. [16] K. Punnonen, P. Autio, U. Kiistala, and M. Ahotupa, “In-vivo
effects of solar-simulated ultraviolet irradiation on antioxidant
enzymes and lipid peroxidation in human epidermis,” British
Journal of Dermatology, vol. 125, no. 1, pp. 18–20, 1991. [33] J.-P. Borel, J.-C. Monboisse, and G. Bellon, “Inflammation,
collagène et radicaux libres oxygénés,” Médecine Sciences,
vol. 4, no. 5, pp. 304–310, 1988. [17] C. W. Trenam, A. J. Dabbagh, D. R. Blake, and C. J. Morris,
“The role of iron in an acute model of skin inflammation
induced by reactive oxygen species (ROS),” British Journal of
Dermatology, vol. 126, no. 3, pp. 250–256, 1992. [34] A. G. Gunin, N. K. Kornilova, V. V. Petrov, and O. V. Vasil'eva, “Age-related changes in the number and prolifera-
tion of fibroblasts in the human skin,” Advances in Gerontol-
ogy, vol. 24, no. 1, pp. 43–47, 2011. [18] J. R. Stewart, T. Moore, and B. Bennett, “Effect of free-radical
scavengers and hyperbaric oxygen on random-pattern skin
flaps,” Archives of Surgery, vol. 129, no. 9, pp. 982–988, 1994. [35] E. K. Ahmed, A. Rogowska-Wrzesinska, P. Roepstorff, A. L. Bulteau, and B. Friguet, “Protein modification and replicative
senescence of WI-38 human embryonic fibroblasts,” Aging
Cell, vol. 9, no. 2, pp. 252–272, 2010. [19] M. Lopez-Torres, Y. Shindo, and L. Packer, “Effect of age on
antioxidants and molecular markers of oxidative damage in
murine epidermis and dermis,” Journal of Investigative Der-
matology, vol. 102, no. 4, pp. 476–480, 1994. [36] M. Yaar, M. S. Eller, and B. A. 5. Conclusion and Perspectives Without any intention to overestimate the importance of the
free radical theory of aging over others, it is possible to sug-
gest that observed morphological and ultrastructural changes
in old skin are a consequence of elevated oxidative pressure
and that the relation between the increased markers of oxida-
tive damage and the following compensatory elevation of AD
cannot be excluded. A similar conclusion, but in the other
direction, is valid when it comes to young skin. Even though
the free radical theory of aging is limited in providing us with
all of the answers regarding aging, our study shows that
altered redox equilibrium in skin has a role in the develop-
ment of this process throughout life, despite some assump-
tions that Harman’s theory is overcome or dead [59]. Perhaps, the best question would be, How long would we live
with reduced or no AD? Possibly, just like erythroid progen-
itor cells without manganese SOD, we ended up dying before
maturation [60]. In any case, research should be focused on
this direction, when aging and free radical theory of aging
are in question. [8] T. Horio and H. Okamoto, “Oxygen intermediates are
involved in ultraviolet radiation-induced damage of Langer-
hans cells,” Journal of Investigative Dermatology, vol. 88,
no. 6, pp. 699–702, 1987. [9] R. R. Bowers, J. Harmon, S. Prescott, J. Asano, and S. Wynne,
“Fowl model for vitiligo: genetic regulation on the fate of the
melanocytes,” Pigment Cell & Melanoma Research, vol. 3,
pp. 242–248, 1990. [10] J. B. Sedgwick, P. R. Bergstresser, and E. R. Hurd, “Increased
superoxide generation by normal granulocytes incubated in
sera from patients with psoriasis,” Journal of Investigative Der-
matology, vol. 76, no. 3, pp. 158–163, 1981. [11] G. Michaëlsson and L. E. Edqvist, “Erythrocyte glutathione
peroxidase activity in acne vulgaris and the effect of selenium
and vitamin E treatment,” Acta Dermato-Venereologica,
vol. 64, no. 1, pp. 9–14, 1984. [12] I. O. Aruoma and B. Halliwell, “DNA damage and free radi-
cals,” Chemistry in Britain, vol. 35, pp. 149–152, 1991. [13] J. B. Rotstein and T. J. Slaga, “Effect of exogenous glutathione
on
tumor
progression
in
the
murine
skin
multistage Oxidative Medicine and Cellular Longevity 11 glutathione peroxidase activity,” Gerontology, vol. 50, no. 4,
pp. 193–199, 2004. carcinogenesis model,” Carcinogenesis, vol. 9, no. 9, pp. 1547–
1551, 1988. [30] I. Emerit, “Free radical and aging of the skin,” in Free Radicals
and Aging, I. 5. Conclusion and Perspectives Wang et al., “Increase of
oxidatively modified protein is associated with a decrease
of proteasome activity and content in aging epidermal
cells,” The Journals of Gerontology. Series A, Biological Sci-
ences and Medical Sciences, vol. 55, no. 5, pp. B220–B227,
2000. [26] M. Luthman and A. Holmgren, “Rat liver thioredoxin and
thioredoxin reductase: purification and characterization,” Bio-
chemistry, vol. 21, no. 26, pp. 6628–6633, 1982. [27] A. Jankovic, I. Golic, M. Markelic et al., “Two key temporally
distinguishable molecular and cellular components of white
adipose tissue browning during cold acclimation,” Journal of
Physiology, vol. 593, no. 15, pp. 3267–3280, 2015. [43] R. J. Kapphahn, B. M. Giwa, K. M. Berg et al., “Retinal
proteins modified by 4-hydroxynonenal: identification of
molecular targets,” Experimental Eye Research, vol. 83,
no. 1, pp. 165–175, 2006. [28] A. Jankovic, C. Ferreri, M. Filipovic et al., “Targeting the
superoxide/nitric oxide ratio by L-arginine and SOD mimic
in diabetic rat skin,” Free Radical Research, vol. 50, Supple-
ment 1, pp. S51–S63, 2016. [44] M. U. Dianzani, “4-Hydroxynonenal and cell signalling,” Free
Radical Research, vol. 28, no. 6, pp. 553–560, 1998. [29] M. Matsuo, H. Ikeda, T. Sugihara, S. Horiike, Y. Okano, and
H. Masaki, “Resistance of cultured human skin fibroblasts
from old and young donors to oxidative stress and their [45] D. R. Petersen
and
J. A. Doorn,
“Reactions
of
4-hydroxynonenal with proteins and cellular targets,” Free
Radical Biology and Medicine, vol. 37, no. 7, pp. 937–945, 2004. Oxidative Medicine and Cellular Longevity 12 12 [46] R. Schaur, W. Siems, N. Bresgen, and P. Eckl, “4-Hydroxy-no-
nenal-a bioactive lipid peroxidation product,” Biomolecules,
vol. 5, no. 4, pp. 2247–2337, 2015. [47] L. E. Rikans and K. R. Hornbrook, “Lipid peroxidation, antiox-
idant protection and aging,” Biochimica et Biophysica Acta
(BBA) - Molecular Basis of Disease, vol. 1362, no. 2-3,
pp. 116–127, 1997. [48] A. B. Salmon, G. Kim, C. Liu et al., “Effects of transgenic
methionine sulfoxide reductase A (MsrA) expression on life-
span and age-dependent changes in metabolic function in
mice,” Redox Biology, vol. 10, pp. 251–256, 2016. [49] M. R. Noh, K. Y. Kim, S. J. Han, J. I. Kim, H. Y. Kim, and K. M. Park, “Methionine sulfoxide reductase A deficiency exacer-
bates cisplatin-induced nephrotoxicity via increased mito-
chondrial damage and renal cell death,” Antioxidants &
Redox Signaling, vol. 27, no. 11, pp. 727–741, 2017. [50] J. Moskovitz, S. Bar-Noy, W. M. Williams, J. 5. Conclusion and Perspectives Requena, B. S. Berlett, and E. R. Stadtman, “Methionine sulfoxide reductase
(MsrA) is a regulator of antioxidant defense and lifespan in
mammals,” Proceedings of the National Academy of Sciences,
vol. 98, no. 23, pp. 12920–12925, 2001. [51] B. C. Lee, A. Dikiy, H. Y. Kim, and V. N. Gladyshev, “Func-
tions and evolution of selenoprotein methionine sulfoxide
reductases,” Biochimica et Biophysica Acta (BBA) - General
Subjects, vol. 1790, no. 11, pp. 1471–1477, 2009. [52] I. Petropoulos, J. Mary, M. Perichon, and B. Friguet, “Rat
peptide methionine sulphoxide reductase: cloning of the
cDNA, and downregulation of gene expression and enzyme
activity during aging,” Biochemical Journal, vol. 355, no. 3,
pp. 819–825, 2001. [53] C. R. Picot, M. Perichon, J. C. Cintrat, B. Friguet, and
I. Petropoulos, “The peptide methionine sulfoxide reductases,
MsrA and MsrB (hCBS-1), are downregulated during replica-
tive senescence of human WI-38 fibroblasts,” FEBS Letters,
vol. 558, no. 1-3, pp. 74–78, 2004. [54] J. L. Pablos, P. E. Carreira, L. Serrano, P. Del Castillo, and J. J. Gomez-Reino, “Apoptosis and proliferation of fibroblasts dur-
ing postnatal skin development and scleroderma in the
tight-skin mouse,” Journal of Histochemistry & Cytochemistry,
vol. 45, no. 5, pp. 711–719, 1997. [55] E. Niki, “Lipid oxidation in the skin,” Free Radical Research,
vol. 49, no. 7, pp. 827–834, 2015. [56] N. Zarkovic, Z. Ilic, M. Jurin, R. J. Schaur, H. Puhl, and
H. Esterbauer, “Stimulation of HeLa cell growth by physiolog-
ical concentrations of 4-hydroxynonenal,” Cell Biochemistry
and Function, vol. 11, no. 4, pp. 279–286, 1993. [57] J. Ruef, G. N. Rao, F. Li et al., “Induction of rat aortic smooth
muscle cell growth by the lipid peroxidation product 4-hydro-
xy-2-nonenal,” Circulation, vol. 97, no. 11, pp. 1071–1078,
1998. [58] L. Milkovic, A. Cipak Gasparovic, and N. Zarkovic, “Over-
view
on
major
lipid
peroxidation
bioactive
factor
4-hydroxynonenal as pluripotent growth-regulating factor,”
Free Radical Research, vol. 49, no. 7, pp. 850–860, 2015. [59] V. N. Gladyshev, “The free radical theory of aging is dead. Long live the damage theory!,” Antioxidants & Redox Signal-
ing, vol. 20, no. 4, pp. 727–731, 2014. [60] D. Marinkovic, X. Zhang, S. Yalcin et al., “Foxo 3 is required
for the regulation of oxidative stress in erythropoiesis,” Journal
of Clinical Investigation, vol. 117, no. 8, pp. 2133–2144, 2007.
|
https://openalex.org/W4245624592
|
https://peerj.com/articles/cs-414v0.2/submission
|
English
| null |
Peer Review #3 of "A systematic review of security and privacy issues in the internet of medical things; the role of machine learning approaches (v0.1)"
| null | 2,021
|
cc-by
| 38,842
|
A systematic review of Security and Privacy issues in the
Internet of Medical Things; The role of Machine Learning
approaches Shilan S. Hameed
Corresp., 1, 2 , Wan H Hassan
1 , Liza Abdul Latiff
3 , Fahad Ghabban
4 1 Malaysia-Japan International Institute of Technology (MJIIT), Universiti Teknologi Malaysia, Kuala Lumpur, Malaysia
2 Directorate of Information Technology, Koya University, Koya, Kurdistan Region, Iraq
3 Razak Faculty of Technology and Informatics, Universiti Teknologi Malaysia, Kuala Lumpur, Malaysia
4 Information Systems Department, College of Computer Sciences and Engineering, Taibah University, Medina, Saudi Arabia, Saudi Arabia
Corresponding Author: Shilan S. Hameed
Email address: hameed.s@graduate.utm.my 1 Malaysia-Japan International Institute of Technology (MJIIT), Universiti Teknologi Malaysia, Kuala Lumpur, Malaysia
2 Directorate of Information Technology, Koya University, Koya, Kurdistan Region, Iraq
3 Razak Faculty of Technology and Informatics, Universiti Teknologi Malaysia, Kuala Lumpur, Malaysia
4 Information Systems Department, College of Computer Sciences and Engineering, Taibah University, Medina, Saudi Arabia, Saudi Arabia
Corresponding Author: Shilan S. Hameed
Email address: hameed.s@graduate.utm.my Corresponding Author: Shilan S. Hameed
Email address: hameed.s@graduate.utm.my Corresponding Author: Shilan S. Hameed
Email address: hameed.s@graduate.utm.my Background . The Internet of Medical Things (IoMTs) is gradually replacing the traditional healthcare
system. However, little attention has been paid to their security requirements in the development of the
IoMT devices and systems. One of the main reasons can be the difficulty of tuning conventional security
solutions to the IoMT system. Machine Learning (ML) has been successfully employed in the attack
detection and mitigation process. Advanced ML technique can also be a promising approach to address
the existing and anticipated IoMT security and privacy issues. However, because of the existing
challenges of IoMT system, it is imperative to know how these techniques can be effectively utilized to
meet the security and privacy requirements without affecting the IoMT systems quality, services, and
device’s lifespan. Methodology. This paper is devoted to perform a Systematic Literature Review (SLR) on the security
and privacy issues of IoMT and their solutions by ML techniques. The recent research papers
disseminated between 2010 to 2020 are selected from multiple databases and a standardized SLR
method is conducted. A total of 153 papers were reviewed and a critical analysis were conducted on the
selected papers. Furthermore, this review study attempts to highlight the limitation of the current
methods and aims to find possible solutions to them. Thus, a detailed analysis was carried out on the
selected papers through focusing on their methods, advantages, limitations, the utilized tools, and data. Results. Computer Science Computer Science Manuscript to be reviewed Computer Science Computer Science Manuscript to be reviewed Manuscript to be reviewed 1 A systematic review of Sec
2 the Internet of Medical Thin
3 Learning approaches
4
5
Shilan S. Hameed 1,2,*, Wan H Hassan1, Liza A
6
7
1 Malaysia-Japan International Institute of Tech
8
Kuala Lumpur, Malaysia. 9
2 Directorate of Information Technology, Koya
10
3 Razak Faculty of Technology and Informatics
11
Malaysia. 12
4 Information Systems Department, College of
13
University, Medina, Saudi Arabia
14
15
Corresponding author:
16
Shilan S. Hameed
17
UTM KL, Jalan Sultan Yahya Petra, 54100 Ku
18
Email address: hameed.s@graduate.utm.my
19
20
Abstract
21
Background. The Internet of Medical Things (
22
healthcare system. However, little attention has
23
development of the IoMT devices and systems
24
tuning conventional security solutions to the Io
25
successfully employed in the attack detection a
26
can also be a promising approach to address the
27
privacy issues. However, because of the existin
28
know how these techniques can be effectively u
29
requirements without affecting the IoMT system
30
Methodology. This paper is devoted to perform
31
security and privacy issues of IoMT and their s
32
papers disseminated between 2010 to 2020 are
33
standardized SLR method is conducted. A total
34
analysis were conducted on the selected papers
35
highlight the limitation of the current methods
36
a detailed analysis was carried out on the select
37
advantages, limitations, the utilized tools, and d
Computer Science 1 A systematic review of Security and Privacy issues in
2 the Internet of Medical Things; The role of Machine
3 Learning approaches
4
5
Shilan S. Hameed 1,2,*, Wan H Hassan1, Liza Abdul Latiff3 , Fahad Ghabban4
6
7
1 Malaysia-Japan International Institute of Technology (MJIIT), University Teknologi Malaysia,
8
Kuala Lumpur, Malaysia. 9
2 Directorate of Information Technology, Koya University, Koya, Kurdistan Region-F.R., Iraq. 10
3 Razak Faculty of Technology and Informatics, University Teknologi Malaysia, Kuala Lumpur,
11
Malaysia. 12
4 Information Systems Department, College of Computer Sciences and Engineering, Taibah
13
University, Medina, Saudi Arabia
14
15
Corresponding author:
16
Shilan S. Hameed
17
UTM KL, Jalan Sultan Yahya Petra, 54100 Kuala Lumpur, Malaysia. 18
Email address: hameed.s@graduate.utm.my
19
20
Abstract
21
Background. The Internet of Medical Things (IoMTs) is gradually replacing the traditional
22
healthcare system. Computer Science However, little attention has been paid to their security requirements in the
23
development of the IoMT devices and systems. One of the main reasons can be the difficulty of
24
tuning conventional security solutions to the IoMT system. Machine Learning (ML) has been
25
successfully employed in the attack detection and mitigation process. Advanced ML technique
26
can also be a promising approach to address the existing and anticipated IoMT security and
27
privacy issues. However, because of the existing challenges of IoMT system, it is imperative to
28
know how these techniques can be effectively utilized to meet the security and privacy
29
requirements without affecting the IoMT systems quality, services, and device’s lifespan. 30
Methodology. This paper is devoted to perform a Systematic Literature Review (SLR) on the
31
security and privacy issues of IoMT and their solutions by ML techniques. The recent research
32
papers disseminated between 2010 to 2020 are selected from multiple databases and a
33
standardized SLR method is conducted. A total of 153 papers were reviewed and a critical
34
analysis were conducted on the selected papers. Furthermore, this review study attempts to
35
highlight the limitation of the current methods and aims to find possible solutions to them. Thus,
36
a detailed analysis was carried out on the selected papers through focusing on their methods,
37
advantages, limitations, the utilized tools, and data. 1 A systematic review of Security and P
2 the Internet of Medical Things; The rol
3 Learning approaches
4
5
Shilan S. Hameed 1,2,*, Wan H Hassan1, Liza Abdul Latiff3 , Fahad G
6
7
1 Malaysia-Japan International Institute of Technology (MJIIT), Un
8
Kuala Lumpur, Malaysia. 9
2 Directorate of Information Technology, Koya University, Koya, K
10
3 Razak Faculty of Technology and Informatics, University Teknolo
11
Malaysia. 12
4 Information Systems Department, College of Computer Sciences a
13
University, Medina, Saudi Arabia
14
15
Corresponding author:
16
Shilan S. Hameed
17
UTM KL, Jalan Sultan Yahya Petra, 54100 Kuala Lumpur, Malaysi
18
Email address: hameed.s@graduate.utm.my
19
20
Abstract
21
Background. The Internet of Medical Things (IoMTs) is gradually
22
healthcare system. However, little attention has been paid to their se
23
development of the IoMT devices and systems. One of the main rea
24
tuning conventional security solutions to the IoMT system. Computer Science Machin
25
successfully employed in the attack detection and mitigation proces
26
can also be a promising approach to address the existing and anticip
27
privacy issues. However, because of the existing challenges of IoMT
28
know how these techniques can be effectively utilized to meet the se
29
requirements without affecting the IoMT systems quality, services,
30
Methodology. This paper is devoted to perform a Systematic Litera
31
security and privacy issues of IoMT and their solutions by ML techn
32
papers disseminated between 2010 to 2020 are selected from multip
33
standardized SLR method is conducted. A total of 153 papers were
34
analysis were conducted on the selected papers. Furthermore, this re
35
highlight the limitation of the current methods and aims to find poss
36
a detailed analysis was carried out on the selected papers through fo
37
advantages, limitations, the utilized tools, and data. A systematic review of Security and Privacy issues in the
Internet of Medical Things; The role of Machine Learning
approaches It was observed that ML techniques have been significantly deployed for device and network
layer security. Most of the current studies improved traditional metrics while ignored performance
complexity metrics in their evaluations. Their studies environments and utilized data barely represent
IoMT system. Therefore, conventional ML techniques may fail if metrics such as resource complexity and
power usage are not considered. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science 1 A systematic review of Security and Privacy issues in
2 the Internet of Medical Things; The role of Machine
3 Learning approaches 5
Shilan S. Hameed 1,2,*, Wan H Hassan1, Liza Abdul Latiff3 , Fahad Ghabban4 9
2 Directorate of Information Technology, Koya University, Koya, Kurdistan Region-F.R., Iraq. 10
3 Razak Faculty of Technology and Informatics, University Teknologi Malaysia, Kuala Lumpur,
11
Malaysia. Manuscript to be reviewed 38
Results. It was observed that ML techniques have been significantly deployed for device and
39
network layer security. Most of the current studies improved traditional metrics while ignored
40
performance complexity metrics in their evaluations. Their studies environments and utilized
41
data barely represent IoMT system. Therefore, conventional ML techniques may fail if metrics
42
such as resource complexity and power usage are not considered. 43
44
Keywords: The Internet of Medical Things, IoMT, Security and Privacy, Machine Learning
45
Background
46
A. IoMT and its Classification
47
The Internet of Thing (IoT) is a fast growing technology by which the infrastructures,
48
computerized machines, physical things, applications and individuals are allowed to connect,
49
communicate, capture and exchange information through networking (Farahani et al. 2018; Noor
50
& Hassan 2019). Hence, the Internet of Medical Thing (IoMT) is the application of IoT in
51
medicine and healthcare industry (Alsubaei et al. 2019a; He et al. 2018). It is anticipated that
52
with the implementation of IoMT a significant improvement in the efficiency and standard of
53
treatment is achieved thanks to the steady innovations in IoT including the development in
54
microprocessors, biosensor architecture and evolving 5G technologies (Ahad et al. 2019). 55
However, it is hard to maintain a specific architecture as a baseline due to the variety of devices
56
and their different usage. Consequently, various approaches for the IoT architectures and layers
57
were presented in literature (Sethi & Sarangi 2017; Weyrich & Ebert 2015) such as protocol-
58
based architecture (Burhan et al. 2018; Mosenia & Jha 2016; Weyrich & Ebert 2015) and data
59
processing stages known as edge, fog, and cloud based approaches (Escamilla-Ambrosio et al. 60
2018). In the current review, we present two common architectures of the IoMT. In the first
61
architecture, the IoMT is treated as to be composed of three main elements: (i) network of
62
wireless body sensor, which is device layer, (ii) internet connected smart access points
63
(gateway), which is fog layer, and (iii) cloud computing and big data service, which is cloud
64
layer (Rahmani et al. 2018). The second architecture is simply defined by sensing/device layer,
65
network layer, and server (personal/medical) layer (Sun et al. 2019). 20
Abstract 20
Abstract
21
Background. The Internet of Medical Things (IoMTs) is gradually replacing the traditional
22
healthcare system. However, little attention has been paid to their security requirements in the
23
development of the IoMT devices and systems. One of the main reasons can be the difficulty of
24
tuning conventional security solutions to the IoMT system. Machine Learning (ML) has been
25
successfully employed in the attack detection and mitigation process. Advanced ML technique
26
can also be a promising approach to address the existing and anticipated IoMT security and
27
privacy issues. However, because of the existing challenges of IoMT system, it is imperative to
28
know how these techniques can be effectively utilized to meet the security and privacy
29
requirements without affecting the IoMT systems quality, services, and device’s lifespan. 30
Methodology. This paper is devoted to perform a Systematic Literature Review (SLR) on the
31
security and privacy issues of IoMT and their solutions by ML techniques. The recent research
32
papers disseminated between 2010 to 2020 are selected from multiple databases and a
33
standardized SLR method is conducted. A total of 153 papers were reviewed and a critical
34
analysis were conducted on the selected papers. Furthermore, this review study attempts to
35
highlight the limitation of the current methods and aims to find possible solutions to them. Thus,
36
a detailed analysis was carried out on the selected papers through focusing on their methods,
37
advantages, limitations, the utilized tools, and data. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed The second architectu
65
network layer, and server (personal/medical) layer
66
have divided the IoMT architecture into more than
67
adopted for a specific application of IoMT (Elrawy
68
Furthermore, there are four types of smart medical
69
their location of implementation on human body (N
70
brief explanation on the types of IoMT devices and
71
and Drug Administration (FDA), the IoMT devices
72
value. Implantable devices such as Implantable car
73
Electrocardiography (ECG), Electromyography (EM
74
under high risk category Hence they are regulated 38
Results. It was observed that ML techniques have been significantly deployed for device and
39
network layer security. Most of the current studies improved traditional metrics while ignored
40
performance complexity metrics in their evaluations. Their studies environments and utilized
41
data barely represent IoMT system. Therefore, conventional ML techniques may fail if metrics
42
such as resource complexity and power usage are not considered. 3 46
A. IoMT and its Classification
47
The Internet of Thing (IoT) is a fast growing technology by which the infrastructures,
48
computerized machines, physical things, applications and individuals are allowed to connect,
49
communicate, capture and exchange information through networking (Farahani et al. 2018; Noor
50
& Hassan 2019). Hence, the Internet of Medical Thing (IoMT) is the application of IoT in
51
medicine and healthcare industry (Alsubaei et al. 2019a; He et al. 2018). It is anticipated that
52
with the implementation of IoMT a significant improvement in the efficiency and standard of
53
treatment is achieved thanks to the steady innovations in IoT including the development in
54
microprocessors, biosensor architecture and evolving 5G technologies (Ahad et al. 2019). 55
However, it is hard to maintain a specific architecture as a baseline due to the variety of devices
56
and their different usage. Consequently, various approaches for the IoT architectures and layers
57
were presented in literature (Sethi & Sarangi 2017; Weyrich & Ebert 2015) such as protocol-
58
based architecture (Burhan et al. 2018; Mosenia & Jha 2016; Weyrich & Ebert 2015) and data
59
processing stages known as edge, fog, and cloud based approaches (Escamilla-Ambrosio et al. 60
2018). In the current review, we present two common architectures of the IoMT. Manuscript to be reviewed However, some researchers
66
have divided the IoMT architecture into more than three layers based on the requirements
67
adopted for a specific application of IoMT (Elrawy et al. 2018; Grammatikis et al. 2019). 68
Furthermore, there are four types of smart medical devices in IoMT that are categorized based on
69
their location of implementation on human body (Nanayakkara et al. 2019). Table 1 shows a
70
brief explanation on the types of IoMT devices and their properties. According to the U.S. Food
71
and Drug Administration (FDA), the IoMT devices can be also classified based on their risk
72
value. Implantable devices such as Implantable cardioverter-defibrillator (ICD),
73
Electrocardiography (ECG), Electromyography (EMG), and Electroencephalography (EEG) are
74
under high-risk category. Hence, they are regulated and certified by FDA. However, non- 38
Results. It was observed that ML techniques have
39
network layer security. Most of the current studies
40
performance complexity metrics in their evaluation
41
data barely represent IoMT system. Therefore, con
42
such as resource complexity and power usage are n
43
44
Keywords: The Internet of Medical Things, IoMT
45
Background
46
A. IoMT and its Classification
47
The Internet of Thing (IoT) is a fast growing techn
48
computerized machines, physical things, applicatio
49
communicate, capture and exchange information th
50
& Hassan 2019). Hence, the Internet of Medical Th
51
medicine and healthcare industry (Alsubaei et al. 2
52
with the implementation of IoMT a significant imp
53
treatment is achieved thanks to the steady innovatio
54
microprocessors, biosensor architecture and evolvi
55
However, it is hard to maintain a specific architectu
56
and their different usage. Consequently, various ap
57
were presented in literature (Sethi & Sarangi 2017;
58
based architecture (Burhan et al. 2018; Mosenia &
59
processing stages known as edge, fog, and cloud ba
60
2018). In the current review, we present two comm
61
architecture, the IoMT is treated as to be composed
62
wireless body sensor, which is device layer, (ii) int
63
(gateway), which is fog layer, and (iii) cloud comp
64
layer (Rahmani et al. 2018). 79
The types and characteristics of medical devices used in the IoMT system 99
The data analytics are conventionally performed at the cloud. However, cloud computing faces
100
the issues of delay and privacy. For this reason, the term of fog computing at gateways (fog
101
devices) was introduced in 2017 (Rahmani et al. 2018). This approach is used to shift some of
102
the cloud computing works closer to the smart devices aiming at achieving a faster computation
103
with keeping privacy (Dang et al. 2019; Firouzi et al. 2018; Rahmani et al. 2018). It is not
104
deniable that using smart medical devices has made the life easier and healthier. However, there
105
are many safety and security gaps in these devices that put not only the devices at risk but expose
106
a significant risk to the patient’s life (Yaacoub et al. 2020) Manuscript to be reviewed 79
The types and characteristics of medical devices used in the IoMT system. 80
81
Medical devices and biosensors are responsible for capturing body’s vital signs, and transferring
82
a huge real-time raw biological data such as heart rate, brain signal, temperature of the body, and
83
glucose level in blood (Dang et al. 2019). These raw data are accumulated and processed at the
84
personal servers that are either devices located near the patient’s body such as mobile phones,
85
medical programmers, and laptops or devices placed far from body such as gateways and routers. 86
Additionally, personal servers usually include a computing analysis facility, which is coupled
87
with a local archiving database in order to store the initial records of the patient. Furthermore, its
88
warning system alerts the patient whenever an abnormality is observed (Newaz et al. 2020). The
89
communication link between body area network (sensors attached to or close to human body)
90
and personal server is usually employed by low-power wireless networking technologies such as
91
Bluetooth or Bluetooth Low Energy (BLE), Near-Field Communication (NFC), Radio-
92
Frequency IDentification (RFID), Zigbee and Z-Wave. Noteworthy, Bluetooth is mostly used in
93
wearable devices, while RFID and NFC are accommodating an ultra-low energy and short-range
94
communication topology. Hence, they are mostly applicable in the implantable devices (Newaz
95
et al. 2020; Sun et al. 2019). In this way, the aggregated data at the personal server is directed to
96
the medical server. Long-range wireless technologies such as Wi-Fi, GSM and LoRa are also
97
used for the connection between the personal server gateway and medical sever. 98 79
The types and characteristics of medical devices used in the IoMT system. 80
81
Medical devices and biosensors are responsible for capturing body’s vital signs, and transferring
82
a huge real-time raw biological data such as heart rate, brain signal, temperature of the body, and
83
glucose level in blood (Dang et al. 2019). These raw data are accumulated and processed at the
84
personal servers that are either devices located near the patient’s body such as mobile phones,
85
medical programmers, and laptops or devices placed far from body such as gateways and routers. 86
Additionally, personal servers usually include a computing analysis facility, which is coupled
87
with a local archiving database in order to store the initial records of the patient. Manuscript to be reviewed Furthermore, its
88
warning system alerts the patient whenever an abnormality is observed (Newaz et al. 2020). The
89
communication link between body area network (sensors attached to or close to human body)
90
and personal server is usually employed by low-power wireless networking technologies such as
91
Bluetooth or Bluetooth Low Energy (BLE), Near-Field Communication (NFC), Radio-
92
Frequency IDentification (RFID), Zigbee and Z-Wave. Noteworthy, Bluetooth is mostly used in
93
wearable devices, while RFID and NFC are accommodating an ultra-low energy and short-range
94
communication topology. Hence, they are mostly applicable in the implantable devices (Newaz
95
et al. 2020; Sun et al. 2019). In this way, the aggregated data at the personal server is directed to
96
the medical server. Long-range wireless technologies such as Wi-Fi, GSM and LoRa are also
97
used for the connection between the personal server gateway and medical sever. 98 Manuscript to be reviewed In the first
61
architecture, the IoMT is treated as to be composed of three main elements: (i) network of
62
wireless body sensor, which is device layer, (ii) internet connected smart access points
63
(gateway), which is fog layer, and (iii) cloud computing and big data service, which is cloud
64
layer (Rahmani et al. 2018). The second architecture is simply defined by sensing/device layer,
65
network layer, and server (personal/medical) layer (Sun et al. 2019). However, some researchers
66
have divided the IoMT architecture into more than three layers based on the requirements
67
adopted for a specific application of IoMT (Elrawy et al. 2018; Grammatikis et al. 2019). 68
Furthermore, there are four types of smart medical devices in IoMT that are categorized based on
69
their location of implementation on human body (Nanayakkara et al. 2019). Table 1 shows a
70
brief explanation on the types of IoMT devices and their properties. According to the U.S. Food
71
and Drug Administration (FDA), the IoMT devices can be also classified based on their risk
72
value. Implantable devices such as Implantable cardioverter-defibrillator (ICD),
73
Electrocardiography (ECG), Electromyography (EMG), and Electroencephalography (EEG) are
74
under high-risk category. Hence, they are regulated and certified by FDA. However, non-
75
implantable devices such as fitness trackers, and smart watches are under low-risk category. 76
Therefore, they neither require certification nor regulation by the FDA (Jaigirdar et al. 2019). PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) 108
B. Security and Privacy in the IoMT 109
The IoMT devices are vulnerable to traditional and zero-day attacks. This is mainly due to the
110
lack of existing security protocols and measures in the fabrication of the devices in addition to
111
the nature of the devices and IoT network. The devices are very tiny, where their computational
112
resources and batteries cannot handle computation of cryptographic and existing heavy security
113
measures. Furthermore, the network of IoMT is heterogeneous, which is composed of different
114
protocols at each layer, making one security solution not applicable to all devices. It was
115
reported in an estimation made by the Statista (The Statistics Portal) that the number of medical
116
IoT devices in the European Union (EU) could reach 25.8 million devices by 2025 (Schneble &
117
Thamilarasu 2019b). In addition to a steady increase in the number of smart medical devices and
118
benefits from the low price of wireless sensors, the security and privacy issues are become the
119
main concern (He et al. 2018). Besides, when internet connected devices are increased, their PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 120
produced data will eventually increase (Dimitrov 2016; Firouzi et al. 2018; Ma et al. 2017). It
121
was predicted by Statista that IoT devices could produce about 79.4 zettabytes (ZB) by 2025 (S. 122
O'Dea 2020). It is well known that not only the IoMT devices are at risk of cyber-attack, their
123
data are also at a high risk. In fact, privacy issues and data disclosure are the current utmost
124
issues in IoMT infrastructure (Gupta et al. 2020b; Xu et al. 2020).. 120
produced data will eventually increase (Dimitrov 2016; Firouzi et al. 2018; Ma et al. 2017). It
121
was predicted by Statista that IoT devices could produce about 79.4 zettabytes (ZB) by 2025 (S. 122
O'Dea 2020). It is well known that not only the IoMT devices are at risk of cyber-attack, their
123
data are also at a high risk. In fact, privacy issues and data disclosure are the current utmost
124
issues in IoMT infrastructure (Gupta et al. 2020b; Xu et al. 2020).. 125
The IoMT security and privacy requirements are different from the traditional network
126
requirements which are usually referred to as CIA-triad (confidentiality, integrity, and
127
availability). Other metrics such as privacy, and non-repudiation are also essential for the IoMT
128
system (Grammatikis et al. 2019; Nanayakkara et al. 2019; Newaz et al. 2020; Yaacoub et al. 129
2020). The followings are definitions of these metrics used in IoMT system. 125
The IoMT security and privacy requirements are different from the traditional network
126
requirements which are usually referred to as CIA-triad (confidentiality, integrity, and
127
availability). Other metrics such as privacy, and non-repudiation are also essential for the IoMT
128
system (Grammatikis et al. 2019; Nanayakkara et al. 2019; Newaz et al. 2020; Yaacoub et al. 129
2020). The followings are definitions of these metrics used in IoMT system. 130
a)
Confidentiality: It limits unauthorized access to certain information and it guarantees the
131
protection of confidential information. Unauthorized access may lead to data leakage and
132
sometimes life-threatening situations (Alassaf & Gutub 2019). 133
b)
Integrity: It ensures that the data and reading of devices are not altered, deleted, or
134
injected by unauthorized parties. Attacks on integrity such as false data injection on implantable
135
pacemaker may lead to death (Yaacoub et al. 2020). Manuscript to be reviewed 136
c)
Availability: It ensures that the data, computing elements, and communications are
137
accessible and working continuously when they are required by a service. System service
138
interruption poses danger on patients health, considering a surgery room equipped with wireless
139
medical devices (Sun et al. 2019). 136
c)
Availability: It ensures that the data, computing elements, and communications are
137
accessible and working continuously when they are required by a service. System service
138
interruption poses danger on patients health, considering a surgery room equipped with wireless
139
medical devices (Sun et al. 2019). 140
d)
Privacy: It ensures that privacy rules are enforced by the IoMT system and allows users
141
to access their private details (Mosenia & Jha 2016). The collection of confidential health
142
records must conform with legal and ethical laws of privacy such as the General Data Protection 144
(Gupta et al. 2020b; Tovino 2016). These rules ensure the protection of private patient data from
145
disclosure. It is important that Electronic Health Records (EHRs) should not be kept beyond the
146
time of their need. Data should not be breached while stored, transmitted, and used (Hassija et al. 147
2019). Due to open communication channels and neglecting data privacy, the possibility of
148
confidential and private details being released, hacked, or compromised is extremely high. This
149
can be compromised through passive attacks or active attacks (Yaacoub et al. 2020). 150
e)
Non-repudiation: This metric encompasses the ability of the system to validate the
151
presence or absence of an activity (Mosenia & Jha 2016; Yeh 2016). It ensures that the sending
152
node is granted a delivery receipt and the receiver node is granted a proof on the identity of the
153
sender so that none of them is denied in the process (Newaz et al. 2020). 154
A threat that risks any of the aforementioned pillars of IoMT security and privacy is considered
155
an attack. Table 2 shows the categorized attacks based on the IoMT targeted layer with respect to
156
their impacts on the security requirements of IoMT system. It is worth to mention that despite of
157
the reported attacks the appearance of zero-day attacks is a daily possibility. While these attacks
158
are threatening the privacy of patients, it is also resulted in unbearable financial damage and loss
159
of reputation (Sun et al. 2019). Manuscript to be reviewed 160
USA, which infected 400 U.S. sites has resulted in ser
161
2020b). According to a latest analysis by Comparitech
162
industry over $160 million since 2016 (Davis 2020a). 163
on brain implants, lead to death (Rathore et al. 2019). 164
attack on the IoMT system, the emergence and develo
165
become a must. 166
167
Table 2:
168
Attacks on different IoMT layers with their respect
169
170
C. Machine Learning Techniques and their
171
Machine Learning (ML) is a branch of Artificial Intell
172
experiences without being explicitly programmed (Ku
173
IoMT, especially at computing nodes such as cloud/fo
174
and producing meaningful information. Recently, ML
175
areas of IoT and IoMT (Alimadadi et al. 2020; Ardabi
176
2019; Pannu 2015; Pramanik et al. 2017; Xiao et al. 20
177
ways to tackle the security issues of the IoMT (Cui et
178
2020; Pirbhulal et al. 2019), whereas traditional securi
179
from zero-day attacks, and they are resource extensive
180
techniques can learn from massive generated data of Io
181
(Hussain et al. 2020). Hence, data hungry ML techniqu
182
There are three main types of ML algorithms which ca
183
problems, namely supervised, unsupervised, and sem
184
technique deals with labelled data, in which classes or
185
supervised learning which are classification and regres
186
supervised learning are Support Vector Machine, Deci
187
methods are used for the attack and malware detecti
188
Intrusion Detection System (IDS) (Eskandari et al. 202
189
easy to label the data by human, unsupervised techniq
190
on their similar traits (Schneble & Thamilarasu 2019b
191
nearest Neighbor (KNN), Self-Organizing Map (SOM
192
(Gupta et al. 2020a). These methods are effective for a
193
et al. 2019; Bostani & Sheikhan 2017; Eskandari et al. 194
not every unlabeled data is useful for prediction (Rath
195
overcome this issue by incorporating some labelled da
196
younger ML sub-field, in which some data are labelled
197
(Wolfgang 2011) Also semi-supervised models can b 160
USA, which infected 400 U.S. sites has resulted in service disruption and data disclosure
161
2020b). According to a latest analysis by Comparitech, these attacks have cost the health
162
industry over $160 million since 2016 (Davis 2020a). Additionally, it was claimed that a
163
on brain implants, lead to death (Rathore et al. 2019). Manuscript to be reviewed Therefore, with the increased risk o
164
attack on the IoMT system, the emergence and development of robust security solutions
165
become a must. 166
167
Table 2:
168
Attacks on different IoMT layers with their respective impact on security requirem
169
170
C. Machine Learning Techniques and their Evaluation Metrics
171
Machine Learning (ML) is a branch of Artificial Intelligence (AI) which learns from data
172
experiences without being explicitly programmed (Kubat 2017). ML can play a vital role
173
IoMT, especially at computing nodes such as cloud/fog computing, for continuous data a
174
and producing meaningful information. Recently, ML has been interestingly applied in v
175
areas of IoT and IoMT (Alimadadi et al. 2020; Ardabili et al. 2020; Cui et al. 2018; Durg
176
2019; Pannu 2015; Pramanik et al. 2017; Xiao et al. 2018). ML can be effectively used in
177
ways to tackle the security issues of the IoMT (Cui et al. 2018; Gupta et al. 2020a; Pande
178
2020; Pirbhulal et al. 2019), whereas traditional security measures cannot prevent the sys
179
from zero-day attacks, and they are resource extensive for the IoMT systems. Advanced M
180
techniques can learn from massive generated data of IoMT, thereby finding new trends o
181
(Hussain et al. 2020). Hence, data hungry ML techniques can be perfectly tuned with IoM
182
There are three main types of ML algorithms which can be used for solving the IoMT sec
183
problems, namely supervised, unsupervised, and semi-supervised algorithms. The Sup
184
technique deals with labelled data, in which classes or labels are known. There are two fo
185
supervised learning which are classification and regression (Kubat 2017). Examples of
186
supervised learning are Support Vector Machine, Decision Trees and Neural Network. Th
187
methods are used for the attack and malware detection purpose such as signature based
188
Intrusion Detection System (IDS) (Eskandari et al. 2020; RM et al. 2020). Since it is not
189
easy to label the data by human, unsupervised techniques are utilized to categorize them
190
on their similar traits (Schneble & Thamilarasu 2019b). Examples of these techniques are
191
nearest Neighbor (KNN), Self-Organizing Map (SOM), and Latent Dirichlet Allocation (
192
(Gupta et al. 2020a). These methods are effective for anomaly detection of new attacks
193
et al. 2019; Bostani & Sheikhan 2017; Eskandari et al. 2020). Manuscript to be reviewed For example, a recent ransomware attack on hospitals in the PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed Based on the no free lunch
194
not every unlabeled data is useful for prediction (Rathore & Park 2018). Hence, one can
195
overcome this issue by incorporating some labelled data. Semi-Supervised Learning is a
196
younger ML sub-field, in which some data are labelled out of a wide number of learning 160
USA, which infected 400 U.S. sites has resulted in service disruption and data disclosure (Davis
161
2020b). According to a latest analysis by Comparitech, these attacks have cost the healthcare
162
industry over $160 million since 2016 (Davis 2020a). Additionally, it was claimed that attacks
163
on brain implants, lead to death (Rathore et al. 2019). Therefore, with the increased risk of cyber-
164
attack on the IoMT system, the emergence and development of robust security solutions are
165
become a must. 170
C. Machine Learning Techniques and their Evaluation Metrics Semi-Supervised Learning is a
196
younger ML sub-field, in which some data are labelled out of a wide number of learning data
197
(Wolfgang 2011). Also, semi-supervised models can be used for attack detection and to avoid
198
adversarial attacks on ML techniques (Miyato et al. 2018; Rathore & Park 2018). In addition to
199
the main classes of ML, a new branch called Deep learning (DL) has been recently emerged, 170
C. Machine Learning Techniques and their Evaluation Metrics
171
Machine Learning (ML) is a branch of Artificial Intelligence (AI) which learns from data and
172
experiences without being explicitly programmed (Kubat 2017). ML can play a vital role in
173
IoMT, especially at computing nodes such as cloud/fog computing, for continuous data analysis
174
and producing meaningful information. Recently, ML has been interestingly applied in various
175
areas of IoT and IoMT (Alimadadi et al. 2020; Ardabili et al. 2020; Cui et al. 2018; Durga et al. 176
2019; Pannu 2015; Pramanik et al. 2017; Xiao et al. 2018). ML can be effectively used in many
177
ways to tackle the security issues of the IoMT (Cui et al. 2018; Gupta et al. 2020a; Pandey et al. 178
2020; Pirbhulal et al. 2019), whereas traditional security measures cannot prevent the system
179
from zero-day attacks, and they are resource extensive for the IoMT systems. Advanced ML
180
techniques can learn from massive generated data of IoMT, thereby finding new trends of attacks
181
(Hussain et al. 2020). Hence, data hungry ML techniques can be perfectly tuned with IoMT. 182
There are three main types of ML algorithms which can be used for solving the IoMT security
183
problems, namely supervised, unsupervised, and semi-supervised algorithms. The Supervised
184
technique deals with labelled data, in which classes or labels are known. There are two forms of
185
supervised learning which are classification and regression (Kubat 2017). Examples of
186
supervised learning are Support Vector Machine, Decision Trees and Neural Network. These
187
methods are used for the attack and malware detection purpose such as signature based
188
Intrusion Detection System (IDS) (Eskandari et al. 2020; RM et al. 2020). Since it is not always
189
easy to label the data by human, unsupervised techniques are utilized to categorize them based
190
on their similar traits (Schneble & Thamilarasu 2019b). 170
C. Machine Learning Techniques and their Evaluation Metrics 170
C. Machine Learning Techniques and their Evaluation Metrics
171
Machine Learning (ML) is a branch of Artificial Intelligence (AI) which learns from data and
172
experiences without being explicitly programmed (Kubat 2017). ML can play a vital role in
173
IoMT, especially at computing nodes such as cloud/fog computing, for continuous data analysis
174
and producing meaningful information. Recently, ML has been interestingly applied in various
175
areas of IoT and IoMT (Alimadadi et al. 2020; Ardabili et al. 2020; Cui et al. 2018; Durga et al. 176
2019; Pannu 2015; Pramanik et al. 2017; Xiao et al. 2018). ML can be effectively used in many
177
ways to tackle the security issues of the IoMT (Cui et al. 2018; Gupta et al. 2020a; Pandey et al. 178
2020; Pirbhulal et al. 2019), whereas traditional security measures cannot prevent the system
179
from zero-day attacks, and they are resource extensive for the IoMT systems. Advanced ML
180
techniques can learn from massive generated data of IoMT, thereby finding new trends of attacks
181
(Hussain et al. 2020). Hence, data hungry ML techniques can be perfectly tuned with IoMT. 182
There are three main types of ML algorithms which can be used for solving the IoMT security
183
problems, namely supervised, unsupervised, and semi-supervised algorithms. The Supervised
184
technique deals with labelled data, in which classes or labels are known. There are two forms of
185
supervised learning which are classification and regression (Kubat 2017). Examples of
186
supervised learning are Support Vector Machine, Decision Trees and Neural Network. These
187
methods are used for the attack and malware detection purpose such as signature based
188
Intrusion Detection System (IDS) (Eskandari et al. 2020; RM et al. 2020). Since it is not always
189
easy to label the data by human, unsupervised techniques are utilized to categorize them based
190
on their similar traits (Schneble & Thamilarasu 2019b). Examples of these techniques are K-
191
nearest Neighbor (KNN), Self-Organizing Map (SOM), and Latent Dirichlet Allocation (LDA)
192
(Gupta et al. 2020a). These methods are effective for anomaly detection of new attacks (Ahmad
193
et al. 2019; Bostani & Sheikhan 2017; Eskandari et al. 2020). Based on the no free lunch theory,
194
not every unlabeled data is useful for prediction (Rathore & Park 2018). Hence, one can
195
overcome this issue by incorporating some labelled data. 170
C. Machine Learning Techniques and their Evaluation Metrics Examples of these techniques are K-
191
nearest Neighbor (KNN), Self-Organizing Map (SOM), and Latent Dirichlet Allocation (LDA)
192
(Gupta et al. 2020a). These methods are effective for anomaly detection of new attacks (Ahmad
193
et al. 2019; Bostani & Sheikhan 2017; Eskandari et al. 2020). Based on the no free lunch theory,
194
not every unlabeled data is useful for prediction (Rathore & Park 2018). Hence, one can
195
overcome this issue by incorporating some labelled data. Semi-Supervised Learning is a
196
younger ML sub-field, in which some data are labelled out of a wide number of learning data
197
(Wolfgang 2011). Also, semi-supervised models can be used for attack detection and to avoid
198
adversarial attacks on ML techniques (Miyato et al. 2018; Rathore & Park 2018). In addition to
199
the main classes of ML, a new branch called Deep learning (DL) has been recently emerged, PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 200
which is an advanced form of Neural Network. It has several layers of artificial neural network
201
that mimic the working principle of human brain in processing data for detecting objects,
202
recognizing speech, translating languages, and making decisions. DL can perform feature
203
selection/extraction based on its learning process without the need of another method compared
204
to traditional ML techniques that require an additional feature selection (Amanullah et al. 2020). 205
DL comes in different forms, including Convolution Neural Network (CNN), Recurrent Neural
206
Network (RNN), Long-Short Term Memory (LSTM), Auto-Encoders (AE), Boltzmann Machine
207
(BM), Generative Adversarial Networks (GAN), Feed forward Deep Networks (FDN) and Deep
208
Belief Networks (DBN) (Amanullah et al. 2020; Hussain et al. 2020; Saeed et al. 2016). Auto-
209
encoders are usually the best choice of unsupervised learning to be used for anomaly detection
210
(Lopez-Martin et al. 2017).The use of DL for security solutions such as attack detection could be
211
a durable method for slight mutations or new threats due to its ability to extract features
212
(Amanullah et al. 2020; Diro & Chilamkurti 2018; Durga et al. 2019; Ibitoye et al. 2019; Parra et
213
al. 2020; Sollins 2018). On the other hand, the IoMT sensors generate a large amount of data at a
214
fast speed, thereby producing Big data (Cao 2017; Pramanik et al. 2017; Saheb & Izadi 2019)
215
(Marjani et al. 2017). Big data refers to the data that are too large and/or complex to be
216
efficiently handled by conventional technologies and tools (Dimitrov 2016; Hameed et al. 2017;
217
Sadoughi et al. 2020; Vinitha et al. 2018). A big data is usually defined by three key features,
218
known as 3Vs, which are volume, velocity and variety (Amanullah et al. 2020; Russom 2011). It
219
was seen in literature that DL and big data have been used together as DL performs better with
220
increasing the amount of data (Gheisari et al. 2017). Very recently, DL and Big Data were used
221
in combination with new technologies of blockchain and reinforcement learning for security
222
purposes in the industrial IoT systems (Aman et al. 2020; Liang et al. 2020; Liu et al. 2018). 223
Concludingly, the ML methods can be employed in different security solutions such as: 200
which is an advanced form of Neural Network. Manuscript to be reviewed It has several layers of artificial neural network
201
that mimic the working principle of human brain in processing data for detecting objects,
202
recognizing speech, translating languages, and making decisions. DL can perform feature
203
selection/extraction based on its learning process without the need of another method compared
204
to traditional ML techniques that require an additional feature selection (Amanullah et al. 2020). 205
DL comes in different forms, including Convolution Neural Network (CNN), Recurrent Neural
206
Network (RNN), Long-Short Term Memory (LSTM), Auto-Encoders (AE), Boltzmann Machine
207
(BM), Generative Adversarial Networks (GAN), Feed forward Deep Networks (FDN) and Deep
208
Belief Networks (DBN) (Amanullah et al. 2020; Hussain et al. 2020; Saeed et al. 2016). Auto-
209
encoders are usually the best choice of unsupervised learning to be used for anomaly detection
210
(Lopez-Martin et al. 2017).The use of DL for security solutions such as attack detection could be
211
a durable method for slight mutations or new threats due to its ability to extract features
212
(Amanullah et al. 2020; Diro & Chilamkurti 2018; Durga et al. 2019; Ibitoye et al. 2019; Parra et
213
al. 2020; Sollins 2018). On the other hand, the IoMT sensors generate a large amount of data at a
214
fast speed, thereby producing Big data (Cao 2017; Pramanik et al. 2017; Saheb & Izadi 2019)
215
(Marjani et al. 2017). Big data refers to the data that are too large and/or complex to be
216
efficiently handled by conventional technologies and tools (Dimitrov 2016; Hameed et al. 2017;
217
Sadoughi et al. 2020; Vinitha et al. 2018). A big data is usually defined by three key features,
218
known as 3Vs, which are volume, velocity and variety (Amanullah et al. 2020; Russom 2011). It
219
was seen in literature that DL and big data have been used together as DL performs better with
220
increasing the amount of data (Gheisari et al. 2017). Very recently, DL and Big Data were used
221
in combination with new technologies of blockchain and reinforcement learning for security
222
purposes in the industrial IoT systems (Aman et al. 2020; Liang et al. 2020; Liu et al. 2018). Manuscript to be reviewed 223
Concludingly, the ML methods can be employed in different security solutions such as: 223
Concludingly, the ML methods can be employed in different security solut 224
Sensor Anomaly Detection in Medical Devices: Anomaly detection is the process of finding
225
deviations from the normal data. Such anomalies are attributed to the occurrence of phenomena
226
that do not obey a regular process. These irregularities are triggered by abnormal behaviors such
227
as internal assault or false data injection attack (Amanullah et al. 2020), leading to a false reading
228
of the sensors. ML approach in anomaly detection is the most used mechanism for sensor
229
security. Nonetheless, it needs a considerable adoption in order to be implemented in the IoMT
230
(Butun et al. 2015; Hasan et al. 2019). 224
Sensor Anomaly Detection in Medical Devices: Anomaly detection is the process of finding
225
deviations from the normal data. Such anomalies are attributed to the occurrence of phenomena
226
that do not obey a regular process. These irregularities are triggered by abnormal behaviors such
227
as internal assault or false data injection attack (Amanullah et al. 2020), leading to a false reading
228
of the sensors. ML approach in anomaly detection is the most used mechanism for sensor
229
security. Nonetheless, it needs a considerable adoption in order to be implemented in the IoMT
230
(Butun et al. 2015; Hasan et al. 2019). 231
Authentication and Access Control: Authentication is the process of ensuring whether the
232
IoMT user is correct, while authorization is associated with the extent of access given to each
233
individual (patient and healthcare provider)(Aghili et al. 2019b). They are the most effective
234
methods to prevent attacks on confidentiality and integrity of the IoMT data (Aghili et al. 2019b;
235
Nguyen et al. 2019; Yang et al. 2018). Different techniques can be combined in the process of
236
authentication and authorization such as cryptography, ML, and lightweight approaches (Aghili
237
et al. 2019b; Shen et al. 2018; Wu et al. 2018). Details about these methods can be found in (Sun
238
et al. 2019). 231
Authentication and Access Control: Authentication is the process of ensuring whether the
232
IoMT user is correct, while authorization is associated with the extent of access given to each
233
individual (patient and healthcare provider)(Aghili et al. 2019b). Manuscript to be reviewed They are the most effective
234
methods to prevent attacks on confidentiality and integrity of the IoMT data (Aghili et al. 2019b;
235
Nguyen et al. 2019; Yang et al. 2018). Different techniques can be combined in the process of
236
authentication and authorization such as cryptography, ML, and lightweight approaches (Aghili
237
et al. 2019b; Shen et al. 2018; Wu et al. 2018). Details about these methods can be found in (Sun
238
et al. 2019). PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 239
Intrusion and Malware Detection: The most common intrusion detection is network intrusion
240
detection (NIDS) using network traffic analysis. If the log data of sensors and devices are used,
241
becomes host-based intrusion detection (HIDS) (Aluvalu 2020). Both types of intrusion detectio
242
use metrics for finding anomalies (unknown-attacks) and signatures (known-attacks) that do not
243
match with the normal network traffic or sensor data. These detection systems are similar in
244
having an agent for collecting data and a processing unit in the attack detection and reporting th
245
intrusions. However, they are different in: i) Source of data: host-based (Asfaw et al. 2010),
246
network-based (Maleh et al. 2015), hybrid (Ahmad et al. 2019), ii) Method of detection
247
(Abhishek et al. 2018): signature-based (Wang et al. 2018), anomaly-based, and iii) Architectur
248
centralized, and distributed (Anthi et al. 2018; da Costa et al. 2019). Signature based approache
249
can easily detect the known attacks but unable to detect new attacks. On the other hand,
250
anomaly-based IDS find anomalous behavior by learning from current data. Therefore, they can
251
detect new attacks, but they are less accurate and computationally expensive (Arshad et al. 2020
252
Jan et al. 2019; Midi et al. 2017). The recent attacks such as DoS, DDoS and different malware
253
attacks are the most common mitigated attacks by IDSs (Din et al. 2019; Meidan et al. 2018;
254
Roopak et al. 2020). 255
In the process of evaluating ML techniques in terms of accuracy, reliability, and applicability,
256
some special metrics should be considered. Table 3 shows the most used metrics with their
257
definition and formula. 258
259
Table 3:
260
The evaluation metrics used for ML techniques. 261
262
D. IoMT Security and Privacy Solutions using Machine Learning. 263
264
Research Rationale
265
The unique nature of the IoMT system with small size devices, heterogeneous network, and
266
diverse protocols, has made the implementation of traditional security frameworks difficult by
267
the medical companies. This in turn makes the IoMT system suspectable to different attacks. 268
Recent advancement in the techniques and technologies of ML has led to achieve fruitful
269
strategies to tackle the issues of IoMT security. However, IoMT system facing more challenges
270
than ordinary network systems. 271
IoMT devices usually generate a large amount of streaming data (Amanullah et al. 2020). Manuscript to be reviewed The
272
diversity nature of these data along with the limited power and resources of IoMT devices,
273
especially for the implantable medical devices (Sun et al. 2019), impose a high computational
274
burden on the traditional ML techniques, thereby reducing their effective application in the IoM
275
devices. Hence, new strategies are required to apply the ML approaches efficiently. 276
Consequently, understanding the current security and privacy issues of IoMT system with their
277
respective solutions using ML techniques is essential. Furthermore, it is significant to know the
278
effectiveness of current deployed ML techniques and to understand their solution strength 239
Intrusion and Malware Detection: The most common intrusion detection is network intrusion
240
detection (NIDS) using network traffic analysis. If the log data of sensors and devices are used, it
241
becomes host-based intrusion detection (HIDS) (Aluvalu 2020). Both types of intrusion detection
242
use metrics for finding anomalies (unknown-attacks) and signatures (known-attacks) that do not
243
match with the normal network traffic or sensor data. These detection systems are similar in
244
having an agent for collecting data and a processing unit in the attack detection and reporting the
245
intrusions. However, they are different in: i) Source of data: host-based (Asfaw et al. 2010),
246
network-based (Maleh et al. 2015), hybrid (Ahmad et al. 2019), ii) Method of detection
247
(Abhishek et al. 2018): signature-based (Wang et al. 2018), anomaly-based, and iii) Architecture:
248
centralized, and distributed (Anthi et al. 2018; da Costa et al. 2019). Signature based approaches
249
can easily detect the known attacks but unable to detect new attacks. On the other hand,
250
anomaly-based IDS find anomalous behavior by learning from current data. Therefore, they can
251
detect new attacks, but they are less accurate and computationally expensive (Arshad et al. 2020;
252
Jan et al. 2019; Midi et al. 2017). The recent attacks such as DoS, DDoS and different malware
253
attacks are the most common mitigated attacks by IDSs (Din et al. 2019; Meidan et al. 2018;
254
Roopak et al. 2020). D. IoMT Security and Privacy Solutions using Machine Learning. 276
Consequently, understanding the current security and privacy issues of IoMT system with their
277
respective solutions using ML techniques is essential. Furthermore, it is significant to know the
278
effectiveness of current deployed ML techniques and to understand their solution strength PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 279
offered to the challenges of IoMT system so far. We found that little attention has been paid in
280
literature to elaborate on these issues. Therefore, in this work, a Systematic Literature Review
281
(SLR) is presented, in which attempts are made to reveal the strong points and limitations of the
282
works performed in this area followed by establishing robust improvement strategies. As such,
283
three research questions can be generated as follows:
284
RQ1: What is the current state of the art and direction of study in IoMT security using ML? 285
RQ2: What kind of tools and data are used for applying ML techniques in IoMT security? 286
RQ3: How ML techniques are effectively applied and what are their limitations? 287
This study is intended for new researchers in the field, and for those who are keen to know about
288
recent advances and limitations in IoMT security issues followed by their solutions using ML
289
approaches. 290
291
Related Studies
292
In a review performed by Cui et al. (Cui et al. 2018), an overview of ML application in the
293
domain of generic IoT was reported to focus on the main applications of ML and its relevant
294
techniques in IoT. However, the work has partially covered the IoT security solutions with ML. 295
In another study (Tahsien et al. 2020), Tahsien et al. reviewed the architectures of generic IoT
296
and ML-based potential solutions for IoT security. Another comprehensive review on using ML
297
techniques in generic IoT security was also conducted by Hussain et al. (Hussain et al. 2020). 298
The authors discussed major threats to each layer of IoT and reviewed recent works of using ML
299
techniques in securing IoT. Noticeably, the reviewed studies, tools and datasets were not
300
comprehensively elaborated and the IoMT system was not explored. A review study on security
301
and privacy of IoMT was conducted by Sun et al., revealing the security requirements and
302
challenges of IoMT with more focus on authentication and access control (Sun et al. 2019). 303
Moreover, a survey on IoMT security and privacy was carried out by Newaz et al (Newaz et al. 304
2020), in which a detailed discussion was given on the security and privacy threats in healthcare
305
systems. They also presented a subsection on the current solutions for healthcare IoT security. Manuscript to be reviewed 306
On the other hand, a review on IoMT security issues and limitations with details about the
307
attacks and their impact on IoMT was presented, whereas a special attention was paid on
308
lightweight security solutions (Yaacoub et al. 2020). Concludingly, there have been some
309
reviews found in literature about generic IoT security using ML and DL. However, little
310
attention has been paid to ML applications for the IoMT security and privacy. To this end, the
311
current review is intended to address the role of ML technologies in tackling the issues of IoMT
312
security and privacy. That is by carrying out a comprehensive and systematic review on the
313
related works performed in literature. For this reason, it is difficult to compare our work with the
314
existing surveys. However, a fair relevant comparison of the previous reviews with that of the
315
current study is given in Table 4 279
offered to the challenges of IoMT system so far. W
280
literature to elaborate on these issues. Therefore, in
281
(SLR) is presented, in which attempts are made to
282
works performed in this area followed by establish
283
three research questions can be generated as follow
284
RQ1: What is the current state of the art and directi
285
RQ2: What kind of tools and data are used for appl
286
RQ3: How ML techniques are effectively applied
287
This study is intended for new researchers in the fie
288
recent advances and limitations in IoMT security is
289
approaches. 290
291
Related Studies
292
In a review performed by Cui et al. (Cui et al. 2018
293
domain of generic IoT was reported to focus on th
294
techniques in IoT. However, the work has partially
295
In another study (Tahsien et al. 2020), Tahsien et a
296
and ML-based potential solutions for IoT security. 297
techniques in generic IoT security was also conduc
298
The authors discussed major threats to each layer o
299
techniques in securing IoT. Noticeably, the reviewe
300
comprehensively elaborated and the IoMT system
301
and privacy of IoMT was conducted by Sun et al., r
302
challenges of IoMT with more focus on authenticat
303
Moreover, a survey on IoMT security and privacy w
304
2020), in which a detailed discussion was given on
305
systems. 291
Related Studies 292
In a review performed by Cui et al. (Cui et al. 2018), an overview of ML application in the
293
domain of generic IoT was reported to focus on the main applications of ML and its relevant
294
techniques in IoT. However, the work has partially covered the IoT security solutions with ML. 295
In another study (Tahsien et al. 2020), Tahsien et al. reviewed the architectures of generic IoT
296
and ML-based potential solutions for IoT security. Another comprehensive review on using ML
297
techniques in generic IoT security was also conducted by Hussain et al. (Hussain et al. 2020). 298
The authors discussed major threats to each layer of IoT and reviewed recent works of using ML
299
techniques in securing IoT. Noticeably, the reviewed studies, tools and datasets were not
300
comprehensively elaborated and the IoMT system was not explored. A review study on security
301
and privacy of IoMT was conducted by Sun et al., revealing the security requirements and
302
challenges of IoMT with more focus on authentication and access control (Sun et al. 2019). 303
Moreover, a survey on IoMT security and privacy was carried out by Newaz et al (Newaz et al. 304
2020), in which a detailed discussion was given on the security and privacy threats in healthcare
305
systems. They also presented a subsection on the current solutions for healthcare IoT security. 306
On the other hand, a review on IoMT security issues and limitations with details about the
307
attacks and their impact on IoMT was presented, whereas a special attention was paid on
308
lightweight security solutions (Yaacoub et al. 2020). Concludingly, there have been some
309
reviews found in literature about generic IoT security using ML and DL. However, little
310
attention has been paid to ML applications for the IoMT security and privacy. To this end, the
311
current review is intended to address the role of ML technologies in tackling the issues of IoMT
312
security and privacy. That is by carrying out a comprehensive and systematic review on the
313
related works performed in literature. For this reason, it is difficult to compare our work with the
314
existing surveys. However, a fair relevant comparison of the previous reviews with that of the
315
current study is given in Table 4. Manuscript to be reviewed They also presented a subsection on the cu
306
On the other hand, a review on IoMT security issue
307
attacks and their impact on IoMT was presented, w
308
lightweight security solutions (Yaacoub et al. 2020
309
reviews found in literature about generic IoT secur
310
attention has been paid to ML applications for the I
311
current review is intended to address the role of ML
312
security and privacy. That is by carrying out a com
313
related works performed in literature. For this reaso
314
existing surveys. However, a fair relevant comparis
315
current study is given in Table 4 279
offered to the challenges of IoMT system so far. We found that little attention has been paid in
280
literature to elaborate on these issues. Therefore, in this work, a Systematic Literature Review
281
(SLR) is presented, in which attempts are made to reveal the strong points and limitations of the
282
works performed in this area followed by establishing robust improvement strategies. As such,
283
three research questions can be generated as follows: 318
Comparison between this survey and other related surveys. 318
Comparison between this survey and other related surveys. Computer Science Computer Science Manuscript to be reviewed 319
320
321
Survey methodology
322
Research Design
323
In the current work, a systematic research design i
324
questions, searching for relevant papers was starte
325
Web of Science, Springer Link, Scopus, Science D
326
papers were extracted to answer the research quest
327
comprehensively screened and analyzed. Finally, t
328
methods. 329
Database Searching and Research Sele
330
A keyword-based search was applied by using diff
331
Learning, The Internet of Medical Things, Security
332
synonym list was expanded during researching and
333
generated accordingly, as shown in Fig. 1. Boolean
334
combinations were constructed to form all the sear
335
were carried out in April 2020, whereas each RIT
336
databases followed by saving the query results. Th
337
between January 2010 and April 2020. Initially, th
338
searching and if this option was not available, the
339
abstract) was used. The search queries and results
340
5. Noteworthy, a total of 28,155 search results wer
341
titles were then examined for significant and dupli
342
remained. After that, the papers whose mentioned
343
As such, a total of 153 publications were filtered a
344
through the full text of the papers, only 43 papers w
345
conducting a careful review of their contents. The
346
the topics related to the background of the study. T
347
performed equally on the title, abstract, conclusion
348
while the detailed process of the methodology is sh
349
350
Table 5:
351
The searching queries and results achieved fro
352
353
Table 6:
354
The selection criteria applied to the reviewed p
355
356
Fi
1 Th k
d
d i
th R
h I 319
320
321
Survey methodology
322
Research Design
323
In the current work, a systematic research design is generated. After creating a list of resea
324
questions, searching for relevant papers was started from different databases including IEE
325
Web of Science, Springer Link, Scopus, Science Direct. Then, the most specific and relev
326
papers were extracted to answer the research questions. Later on, the selected papers were
327
comprehensively screened and analyzed. Finally, the results were presented using differen
328
methods. 329
Database Searching and Research Selection
330
A keyword-based search was applied by using different forms and combinations of Machi
331
Learning, The Internet of Medical Things, Security and Privacy and their synonyms. Computer Science Then, the
332
synonym list was expanded during researching and a Research Information Template (RIT) was
333
generated accordingly, as shown in Fig. 1. Boolean expressions with all the keyword
334
combinations were constructed to form all the searching possibilities. The literature searches
335
were carried out in April 2020, whereas each RIT query string was checked in the respective
336
databases followed by saving the query results. The period affecting this analysis was justified
337
between January 2010 and April 2020. Initially, the full metadata would have been selected for
338
searching and if this option was not available, the common search choice (keyword, title and
339
abstract) was used. The search queries and results returned by the databases are shown in Table
340
5. Noteworthy, a total of 28,155 search results were returned from all the five databases. The
341
titles were then examined for significant and duplicate papers and a total of 500 papers were
342
remained. After that, the papers whose mentioned IoMT/ML/security and privacy were chosen. 343
As such, a total of 153 publications were filtered and the remainder was removed. After going
344
through the full text of the papers, only 43 papers were selected for critical analysis and
345
conducting a careful review of their contents. The rest of the papers were used while discussing
346
the topics related to the background of the study. The selection criteria given in Table 6 were
347
performed equally on the title, abstract, conclusion, and full paper in all stages of screening,
348
while the detailed process of the methodology is shown in Fig. 2. 349
350
Table 5:
351
The searching queries and results achieved from five different databases. 352
353
Table 6:
354
The selection criteria applied to the reviewed papers. 355
356
Figure 1: The keywords used in the Research Information Template (RIT). 357
358
Figure 2: The search strategy used for selecting the research papers. Computer Science Then
332
synonym list was expanded during researching and a Research Information Template (RIT
333
generated accordingly, as shown in Fig. 1. Boolean expressions with all the keyword
334
combinations were constructed to form all the searching possibilities. The literature search
335
were carried out in April 2020, whereas each RIT query string was checked in the respect
336
databases followed by saving the query results. The period affecting this analysis was just
337
between January 2010 and April 2020. Initially, the full metadata would have been selecte
338
searching and if this option was not available, the common search choice (keyword, title a
339
abstract) was used. The search queries and results returned by the databases are shown in
340
5. Noteworthy, a total of 28,155 search results were returned from all the five databases. T
341
titles were then examined for significant and duplicate papers and a total of 500 papers we
342
remained. After that, the papers whose mentioned IoMT/ML/security and privacy were ch
343
As such, a total of 153 publications were filtered and the remainder was removed. After g
344
through the full text of the papers, only 43 papers were selected for critical analysis and
345
conducting a careful review of their contents. The rest of the papers were used while discu
346
the topics related to the background of the study. The selection criteria given in Table 6 w
347
performed equally on the title, abstract, conclusion, and full paper in all stages of screenin
348
while the detailed process of the methodology is shown in Fig. 2. 349
350
Table 5:
351
The searching queries and results achieved from five different databases. 352 g
323
In the current work, a systematic research design is generated. After creating a list of research
324
questions, searching for relevant papers was started from different databases including IEEE,
325
Web of Science, Springer Link, Scopus, Science Direct. Then, the most specific and relevant
326
papers were extracted to answer the research questions. Later on, the selected papers were
327
comprehensively screened and analyzed. Finally, the results were presented using different
328
methods. 328
methods. 329
Database Searching and Research Selection
330
A keyword-based search was applied by using different forms and combinations of Machine
331
Learning, The Internet of Medical Things, Security and Privacy and their synonyms. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Manuscript to be reviewed Manuscript to be reviewed 359
360
Results and findings
361
In this work, two types of analysis were performed which include a b
362
technical analysis, as discussed in the next subsections. 363
364
Bibliometric Analysis
365
To show the leading countries whose researchers working in the field
366
each individual paper was examined according to the affiliation of au
367
the USA has made 30% of the papers among 21 affiliated countries, a
368
Moreover, it was found that research performed on ML applications i
369
grown steadily from 10 percent in 2015 to a peak value of 80 percent
370
371
372
Figure 3: Geographical distribution of the papers. 373
374
375
Figure 4: Distribution of the papers by year. 376
377
It is worth to mention that the majority of the papers were journal arti
378
the papers, while 21% of the papers were from conferences of the Ins
379
Electronics Engineering (IEEE) and only 6% of the papers were belo
380
in Fig. 5. Furthermore, a significant number of papers (64%) was pub
381
17% was published by Elsevier, as seen in Fig. 6. 382
383
Figure 5: The type of analyzed papers used in the current researc
384
385
Figure 6: Distribution of the papers according to the publishers. 386
387
388
Studies Technical Analysis
389
390
Classification of Studies
391
The vast majority of ML related articles were about supervised ML, w
392
found to report on a combination of supervised and unsupervised ML
393
on unsupervised ML. Deep learning was also used in some of the stu
394
used big data technology. The papers were further categorized based 359
360
Results and findings
361
In this work, two types of analysis were performed which include a bibliometric analysis and
362
technical analysis, as discussed in the next subsections. 363
364
Bibliometric Analysis
365
To show the leading countries whose researchers working in the field of IoMT and its security,
366
each individual paper was examined according to the affiliation of authors. It was observed that
367
the USA has made 30% of the papers among 21 affiliated countries, as shown in Fig. 3. 368
Moreover, it was found that research performed on ML applications in the IoMT security has
369
grown steadily from 10 percent in 2015 to a peak value of 80 percent in 2019, as shown in Fig. 4. 375
Figure 4: Distribution of the papers by year. 377
It is worth to mention that the majority of the papers were journal article, contributing to 73% of
378
the papers, while 21% of the papers were from conferences of the Institute of Electrical
379
Electronics Engineering (IEEE) and only 6% of the papers were belong to book chapters as seen
380
in Fig. 5. Furthermore, a significant number of papers (64%) was published by the IEEE and
381
17% was published by Elsevier, as seen in Fig. 6. 377
It is worth to mention that the majority of the papers were journal article, contributing to 73% of
378
the papers, while 21% of the papers were from conferences of the Institute of Electrical
379
Electronics Engineering (IEEE) and only 6% of the papers were belong to book chapters as seen
380
in Fig. 5. Furthermore, a significant number of papers (64%) was published by the IEEE and
381
17% was published by Elsevier, as seen in Fig. 6. 329
Database Searching and Research Selection 329
Database Searching and Research Selection
330
A keyword-based search was applied by using different forms and combinations of Machine
331
Learning, The Internet of Medical Things, Security and Privacy and their synonyms. Then, the
332
synonym list was expanded during researching and a Research Information Template (RIT) was
333
generated accordingly, as shown in Fig. 1. Boolean expressions with all the keyword
334
combinations were constructed to form all the searching possibilities. The literature searches
335
were carried out in April 2020, whereas each RIT query string was checked in the respective
336
databases followed by saving the query results. The period affecting this analysis was justified
337
between January 2010 and April 2020. Initially, the full metadata would have been selected for
338
searching and if this option was not available, the common search choice (keyword, title and
339
abstract) was used. The search queries and results returned by the databases are shown in Table
340
5. Noteworthy, a total of 28,155 search results were returned from all the five databases. The
341
titles were then examined for significant and duplicate papers and a total of 500 papers were
342
remained. After that, the papers whose mentioned IoMT/ML/security and privacy were chosen. 343
As such, a total of 153 publications were filtered and the remainder was removed. After going
344
through the full text of the papers, only 43 papers were selected for critical analysis and
345
conducting a careful review of their contents. The rest of the papers were used while discussing
346
the topics related to the background of the study. The selection criteria given in Table 6 were
347
performed equally on the title, abstract, conclusion, and full paper in all stages of screening,
348
while the detailed process of the methodology is shown in Fig. 2. 349
350
Table 5: 358
Figure 2: The search strategy used for selecting the research papers. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 370
371
372
Figure 3: Geographical distribution of the papers. 373
374
375
Figure 4: Distribution of the papers by year. 376
377
It is worth to mention that the majority of the papers were journal article, contributing to 73% of
378
the papers, while 21% of the papers were from conferences of the Institute of Electrical
379
Electronics Engineering (IEEE) and only 6% of the papers were belong to book chapters as seen
380
in Fig. 5. Furthermore, a significant number of papers (64%) was published by the IEEE and
381
17% was published by Elsevier, as seen in Fig. 6. 382
383
Figure 5: The type of analyzed papers used in the current research. 384
385
Figure 6: Distribution of the papers according to the publishers. 386
387
388
Studies Technical Analysis
389
390
Classification of Studies
391
The vast majority of ML related articles were about supervised ML, while the rest of papers were
392
found to report on a combination of supervised and unsupervised ML, with few papers to focus
393
on unsupervised ML. Deep learning was also used in some of the studies, while one study has
394
used big data technology. The papers were further categorized based on the type of medical
395
devices intended to get secured. Almost all studies have focused on the security of wearable
396
devices, while few of them elaborated on the security measures for implantable devices. Only 359
360
Results and findings
361
In this work, two types of analysis were perform
362
technical analysis, as discussed in the next subs
363
364
Bibliometric Analysis
365
To show the leading countries whose research
366
each individual paper was examined according
367
the USA has made 30% of the papers among 2
368
Moreover, it was found that research performed
369
grown steadily from 10 percent in 2015 to a pe
370
371
372
Figure 3: Geographical distribution of the p
373
374
375
Figure 4: Distribution of the papers by year. 376
377
It is worth to mention that the majority of the p
378
the papers, while 21% of the papers were from
379
Electronics Engineering (IEEE) and only 6% o
380
in Fig. 5. Furthermore, a significant number of
381
17% was published by Elsevier, as seen in Fig. 364
Bibliometric Analysis 364
Bibliometric Analysis
365
To show the leading countries whose researchers working in the field of IoMT and its security,
366
each individual paper was examined according to the affiliation of authors. It was observed that
367
the USA has made 30% of the papers among 21 affiliated countries, as shown in Fig. 3. 368
Moreover, it was found that research performed on ML applications in the IoMT security has
369
grown steadily from 10 percent in 2015 to a peak value of 80 percent in 2019, as shown in Fig. 4. 370 375
Figure 4: Distribution of the papers by year. 376
377
It is worth to mention that the majority of the papers were journal article, contributing to 73% of
378
the papers, while 21% of the papers were from conferences of the Institute of Electrical
379
Electronics Engineering (IEEE) and only 6% of the papers were belong to book chapters as seen
380
in Fig. 5. Furthermore, a significant number of papers (64%) was published by the IEEE and
381
17% was published by Elsevier, as seen in Fig. 6. 382
383
Figure 5: The type of analyzed papers used in the current research. 384
385
Figure 6: Distribution of the papers according to the publishers. 386
387
388
Studies Technical Analysis
389
390
Classification of Studies
391
The vast majority of ML related articles were about supervised ML, while the rest of papers were
392
found to report on a combination of supervised and unsupervised ML, with few papers to focus
393
on unsupervised ML. Deep learning was also used in some of the studies, while one study has
394
used big data technology. The papers were further categorized based on the type of medical
395
devices intended to get secured. Almost all studies have focused on the security of wearable
396
devices, while few of them elaborated on the security measures for implantable devices. Only
397
one study was found to focus on securing programmer devices. The targeted IoMT layers in most Manuscript to be reviewed 382
383
Figure 5: The type of analyzed papers used i
384
385
Figure 6: Distribution of the papers accordin
386
387
388
Studies Technical Analysis
389
390
Classification of Studies
391
The vast majority of ML related articles were a
392
found to report on a combination of supervised
393
on unsupervised ML. Deep learning was also u
394
used big data technology. The papers were furt
395
devices intended to get secured. Almost all stud
396
devices, while few of them elaborated on the se Manuscript to be reviewed The authors reported an acc
427
98.1% for their system in distinguishing the fake from genuine glucose measuremen
428
reliability of whole framework was improved by 18% in the case of securing one de
429
enhanced by 90% in the case of securing the whole devices. 430
Khan et al. (Khan et al. 2017) proposed a personal server centered (phone based) M
431
based detection mechanism for the detection of multiple intrusions such as forgery
432
data insertions, and data modifications in the ECG data for smart medical devices. T
433
features by DWT method were generated representing a feature set followed by the
434
into sequences. Each sequence’s probability was then calculated. The probability va
435
as a benchmark to decide if any changes have occurred. Analyzed results showed th
436
has a high detection rate with abnormalities of 5% and 10% in the dataset Howeve 398
of the studies were device and sensor layers. Network layer was addressed in some studies, while
399
cloud layer was reported by two paper. Table 7 shows further details on the disseminated studies
400
related to their ML category and types of devices with their targeted layer. 400
related to their ML category and types of devices with their targeted layer. 401
The papers were then classified into different subsections based on the approaches taken to
402
tackle the security issues of the IoMT, as was discussed in the background section. In addition,
403
an in-depth assessment was carried out through a critical analysis of the articles, demonstrating
404
the strong characteristics and limitations of each study. In the following sections, we discuss on
405
the findings of the aforementioned analysis. 385
Figure 6: Distribution of the papers according to the publishers. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) 407
Classification of papers based on ML category, medical device, the IoM 409
Sensor Anomaly Detection for Medical Devices: Herein, the review results of the current
410
subsection related research articles are presented and analyzed, while the highlighted limitations
411
of the studies are given in Table 8. 412
In a work performed by Haque et al. (Haque et al. 2015), a sensor anomaly detection system was
413
proposed to differentiate the true from false alarms. The research used a historic data to be
414
compared with the actual sensed data for prediction, whereas majority voting was used for their
415
distinguishing. Consequently, the error was calculated based on dynamic threshold. The
416
proposed method has been implemented in Java environment, supplied by the SMO regression. 417
The results illustrated that the proposed system had a high Detection Rate and low FPR for three
418
medical datasets. Furthermore, referring to the security of signals from deep brain stimulators,
419
the authors in (Abdaoui et al. 2020) built a system for distinguishing false alarms from legitimate
420
ones and classified the attacks using Raspberry Pi3 and deep learning. It was found that deep
421
learning can show an accuracy of about 97% to learn and predict the fake signals. Also, a web-
422
based application was generated using the web engine (Flask) for that purpose. pp
g
g
g
(
)
p
p
423
Despite the effective application of ML algorithms, they are generating high computational
424
overhead on the low-power embedded frameworks. Rathore et al. (Rathore et al. 2018c)
425
presented a neural network based MLP solution embedded on an FPGA chip system for securing
426
insulin pump devices that are used by diabetic patients. The authors reported an accuracy of
427
98.1% for their system in distinguishing the fake from genuine glucose measurements. The
428
reliability of whole framework was improved by 18% in the case of securing one device and
429
enhanced by 90% in the case of securing the whole devices. 430
Khan et al. (Khan et al. 2017) proposed a personal server centered (phone based) Markov model-
431
based detection mechanism for the detection of multiple intrusions such as forgery attacks, false
432
data insertions, and data modifications in the ECG data for smart medical devices. The extracted
433
features by DWT method were generated representing a feature set followed by their division
434
into sequences. Each sequence’s probability was then calculated. Manuscript to be reviewed 398
of the studies were device and sensor layers. Network layer was addressed in some
399
cloud layer was reported by two paper. Table 7 shows further details on the dissemi
400
related to their ML category and types of devices with their targeted layer. 401
The papers were then classified into different subsections based on the approaches
402
tackle the security issues of the IoMT, as was discussed in the background section. 403
an in-depth assessment was carried out through a critical analysis of the articles, de
404
the strong characteristics and limitations of each study. In the following sections, w
405
the findings of the aforementioned analysis. 406
Table 7:
407
Classification of papers based on ML category, medical device, the IoMT laye
408
409
Sensor Anomaly Detection for Medical Devices: Herein, the review results of the
410
subsection related research articles are presented and analyzed, while the highlighte
411
of the studies are given in Table 8. 412
In a work performed by Haque et al. (Haque et al. 2015), a sensor anomaly detectio
413
proposed to differentiate the true from false alarms. The research used a historic dat
414
compared with the actual sensed data for prediction, whereas majority voting was u
415
distinguishing. Consequently, the error was calculated based on dynamic threshold. 416
proposed method has been implemented in Java environment, supplied by the SMO
417
The results illustrated that the proposed system had a high Detection Rate and low F
418
medical datasets. Furthermore, referring to the security of signals from deep brain s
419
the authors in (Abdaoui et al. 2020) built a system for distinguishing false alarms fr
420
ones and classified the attacks using Raspberry Pi3 and deep learning. It was found
421
learning can show an accuracy of about 97% to learn and predict the fake signals. A
422
based application was generated using the web engine (Flask) for that purpose. 423
Despite the effective application of ML algorithms, they are generating high compu
424
overhead on the low-power embedded frameworks. Rathore et al. (Rathore et al. 20
425
presented a neural network based MLP solution embedded on an FPGA chip system
426
insulin pump devices that are used by diabetic patients. 407
Classification of papers based on ML category, medical device, the IoM The probability value was used
435
as a benchmark to decide if any changes have occurred. Analyzed results showed that the method
436
has a high detection rate with abnormalities of 5% and 10% in the dataset. However, it has a
437
higher TNR with reduced running time for both 5% and 10% abnormalities. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 438
In another study (Gao & Thamilarasu 2017), some ML techniques were used, including decision
439
tree, SVM and K-means, to detect the security attacks in implantable devices. An external
440
detection device was used to monitor the network and the ML classifiers were utilized to detect
441
anomalies on the gateway device for detecting forced device authentication that results in
442
resource depletion of the device. For this purpose, a feature set specific to IMD devices was
443
constructed. Experimental results demonstrated that decision tree-based algorithms achieved the
444
highest detection accuracy, low false positive rate, fast training and prediction speed compared to
445
those of other algorithms. 446
In another study made by Sehatbakhsh et al. (Sehatbakhsh et al. 2018), SYNDROME was
447
proposed. This method can detect code injection attack in a known program which runs on the
448
system in a real time manner. Statistical based methods such as K-S test and external hardware
449
device were used for detecting signal anomaly. The ability of the method was evaluated by
450
implementing control-flow hijack attacks on a real medical device (syringe pump) embedded
451
system. The evaluation results on using four distinct hardware systems have shown that the
452
proposed model can detect all the attacks with 100% TPR and zero false positive, while the
453
detection latency was less than two milliseconds. 454
In a pioneer work carried out by Zhang et al. (Zhang et al. 2013), a security framework was
455
proposed for medical devices monitoring (MedMon). The proposed model was embedded on an
456
external device which listens to all the passed signals coming from or sending to the medical
457
devices by using a multi-layered anomaly detection (behavioral and physical anomalies). The
458
system is useful for those medical devices that do not use encryption. Consequently, the
459
framework either passively notifies the user or actively jams the signal. This solution does not
460
add power overhead on the medical devices without modification to their software and hardware. 461
An insulin delivery device was tested against the proposed method. Results depicted that the
462
system could successfully detect multiple attacks. 463
For the same purpose, a different approach has been proposed by Newaz et al. (Newaz et al. Computer Science Computer Science Manuscript to be reviewed For the validation purpose, real medical
508
physiological datasets were used. It was concluded that the system was able to effectively detect
509
the anomaly. 510
Additionally, measured data of blood glucose sensor was examined in order to detect adversarial
511
and accidental data modification intrusions (Verner & Butvinik 2017). Here, Otsu’s thresholding
512
algorithm was used with extra statistical analysis to create different informative feature vectors. 513
Then, linear SVM with different misclassification parameters were used to classify the feature
514
vectors. The obtained results on huge patient’s data showed 100% precision and 99.22% recall. 515
This demonstrated that data modification attacks can be effectively detected by utilizing the ML 478
can distinguish between the false and true medical functions. Other researchers (Mohamed et al. 479
2019) attempted to improve the detection efficiency by proposing an intrusion cancellation
480
approach, thereby making the anomaly detection in medical devices efficient. This was achieved
481
by using filters for eliminating noises in the medical data followed by detecting intrusions
482
through statistically analyzed amplitude and frequency. Finally, the detected intrusions were
483
removed to execute the anomaly detection in the medical device for diagnostic purpose. The
484
simulation results applied on two sensor data showed that the system has high TPR and
485
comparable FPR. 486
Anomaly or malfunction sensor reading resulted from defective sensor nodes or produced by
487
fraudulent foreign entities can contribute to medical error and even mortality in patients. 488
Researchers (Salem et al. 2014) suggested a system to identify abnormalities in Wireless Body
489
Area Network (WBAN) for pervasive patient and health surveillance. The proposed software
490
combines advanced data mining and deep learning technologies with existing sensor fusion
491
techniques. The suggested model uses Support Vector Machine (SVM) to identify irregular
492
incidents of the received sensor data. When an anomaly is detected, a regularly updated and
493
regressive prediction model is used to differentiate between the stable and faulty readings
494
resulting in a higher TPR and lower FNR. 495
It was recently (Hau & Lupu 2019) emphasized that identification of fake data injections in low
496
dense Wireless Sensor Networks is important for maintaining the integrity of data, particularly in
497
medical IoT systems. Hence, researchers proposed a framework for detecting false data using
498
anomalies in temporal-attribute correlations between sensor measurements. Manuscript to be reviewed This method is
499
successful in detecting attacks when more than one sensor is colluding to record coherent
500
measurements (Hau & Lupu 2019). Nevertheless, with an increase in the number of colluding
501
sensors, the detection efficiency degrades to a point that the detection fails when most of the
502
sensors are colluding. 503
Furthermore, researchers (Nagdeo & Mahapatro 2019) have implemented an ML model to
504
separate anomalous data from legitimate sensed data. This research used a combination of ANN
505
with Ensemble LinReg as a detection technique for abnormalities in WBAN sensors. Firstly,
506
normal, and abnormal health records were classified. Secondly, regression methodology was
507
used to recognize the anomaly and real vital data. For the validation purpose, real medical
508
physiological datasets were used. It was concluded that the system was able to effectively detect
509
the anomaly. 510
Additionally, measured data of blood glucose sensor was examined in order to detect adversarial
511
and accidental data modification intrusions (Verner & Butvinik 2017). Here, Otsu’s thresholding
512
algorithm was used with extra statistical analysis to create different informative feature vectors. 513
Then, linear SVM with different misclassification parameters were used to classify the feature
514
vectors. The obtained results on huge patient’s data showed 100% precision and 99.22% recall. 515
This demonstrated that data modification attacks can be effectively detected by utilizing the ML
516
approach. 478
can distinguish between the false and true medical functions. Other researchers (Mohamed et al. 479
2019) attempted to improve the detection efficiency by proposing an intrusion cancellation
480
approach, thereby making the anomaly detection in medical devices efficient. This was achieved
481
by using filters for eliminating noises in the medical data followed by detecting intrusions
482
through statistically analyzed amplitude and frequency. Finally, the detected intrusions were
483
removed to execute the anomaly detection in the medical device for diagnostic purpose. The
484
simulation results applied on two sensor data showed that the system has high TPR and
485
comparable FPR 495
It was recently (Hau & Lupu 2019) emphasized that identification of fake data injections in low
496
dense Wireless Sensor Networks is important for maintaining the integrity of data, particularly in
497
medical IoT systems. Hence, researchers proposed a framework for detecting false data using
498
anomalies in temporal-attribute correlations between sensor measurements. Manuscript to be reviewed 464
2019) which is based on ML data-driven security framework, called HealthGuard, for detecting
465
three types of malicious activities in a SHS by considering interconnected body function. Here,
466
ML based techniques (Artificial Neural Network, Decision Tree, Random Forest, and k-Nearest
467
Neighbor) were used to interpret the physiological signs in multiple attached SHS instruments
468
and compare them to identify the differences in the person's body functions, thereby
469
differentiating benevolent and malicious behaviors. Moreover, there is no need to have user
470
identification for the medical devices and the framework does not increase any overhead on the
471
sensors while collecting data. The proposed system is trained with physiological data obtained
472
from eight IoMT devices containing 12 genuine events consist of 7 normal patient activities and
473
five disease associated activities. Results showed an accuracy of 91% and F1 score of 90%. 474
Ben Amor et al. (Ben Amor et al. 2020) suggested an anomaly data detection and separation for
475
the mobile smart healthcare. Two steps were implemented in the study, namely a preprocessing
476
step and a real-time processing step. PCA and Correlation Coefficient were used for feature
477
selection and feature extraction. By this, the system can detect false physiological readings and 463
For the same purpose, a different approach has been proposed by Newaz et al. (Newaz et al. 464
2019) which is based on ML data-driven security framework, called HealthGuard, for detecting
465
three types of malicious activities in a SHS by considering interconnected body function. Here,
466
ML based techniques (Artificial Neural Network, Decision Tree, Random Forest, and k-Nearest
467
Neighbor) were used to interpret the physiological signs in multiple attached SHS instruments
468
and compare them to identify the differences in the person's body functions, thereby
diff
i i
b
l
d
li i
b h
i
h
i
d
h PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 478
can distinguish between the false and true medical functions. Other researchers (Mohamed et al. 479
2019) attempted to improve the detection efficiency by proposing an intrusion cancellation
480
approach, thereby making the anomaly detection in medical devices efficient. This was achieved
481
by using filters for eliminating noises in the medical data followed by detecting intrusions
482
through statistically analyzed amplitude and frequency. Finally, the detected intrusions were
483
removed to execute the anomaly detection in the medical device for diagnostic purpose. The
484
simulation results applied on two sensor data showed that the system has high TPR and
485
comparable FPR. 486
Anomaly or malfunction sensor reading resulted from defective sensor nodes or produced by
487
fraudulent foreign entities can contribute to medical error and even mortality in patients. 488
Researchers (Salem et al. 2014) suggested a system to identify abnormalities in Wireless Body
489
Area Network (WBAN) for pervasive patient and health surveillance. The proposed software
490
combines advanced data mining and deep learning technologies with existing sensor fusion
491
techniques. The suggested model uses Support Vector Machine (SVM) to identify irregular
492
incidents of the received sensor data. When an anomaly is detected, a regularly updated and
493
regressive prediction model is used to differentiate between the stable and faulty readings
494
resulting in a higher TPR and lower FNR. 495
It was recently (Hau & Lupu 2019) emphasized that identification of fake data injections in low
496
dense Wireless Sensor Networks is important for maintaining the integrity of data, particularly in
497
medical IoT systems. Hence, researchers proposed a framework for detecting false data using
498
anomalies in temporal-attribute correlations between sensor measurements. This method is
499
successful in detecting attacks when more than one sensor is colluding to record coherent
500
measurements (Hau & Lupu 2019). Nevertheless, with an increase in the number of colluding
501
sensors, the detection efficiency degrades to a point that the detection fails when most of the
502
sensors are colluding. 503
Furthermore, researchers (Nagdeo & Mahapatro 2019) have implemented an ML model to
504
separate anomalous data from legitimate sensed data. This research used a combination of ANN
505
with Ensemble LinReg as a detection technique for abnormalities in WBAN sensors. Firstly,
506
normal, and abnormal health records were classified. Secondly, regression methodology was
507
used to recognize the anomaly and real vital data. Manuscript to be reviewed This method is
499
successful in detecting attacks when more than one sensor is colluding to record coherent
500
measurements (Hau & Lupu 2019). Nevertheless, with an increase in the number of colluding
501
sensors, the detection efficiency degrades to a point that the detection fails when most of the
502
sensors are colluding. 495
It was recently (Hau & Lupu 2019) emphasized that identification of fake data injections in low
496
dense Wireless Sensor Networks is important for maintaining the integrity of data, particularly in
497
medical IoT systems. Hence, researchers proposed a framework for detecting false data using
498
anomalies in temporal-attribute correlations between sensor measurements. This method is
499
successful in detecting attacks when more than one sensor is colluding to record coherent
500
measurements (Hau & Lupu 2019). Nevertheless, with an increase in the number of colluding
501
sensors, the detection efficiency degrades to a point that the detection fails when most of the
502
sensors are colluding. 503
Furthermore, researchers (Nagdeo & Mahapatro 2019) have implemented an ML model to
504
separate anomalous data from legitimate sensed data. This research used a combination of ANN
505
with Ensemble LinReg as a detection technique for abnormalities in WBAN sensors. Firstly,
506
normal, and abnormal health records were classified. Secondly, regression methodology was
507
used to recognize the anomaly and real vital data. For the validation purpose, real medical
508
physiological datasets were used. It was concluded that the system was able to effectively detect
509
the anomaly. 510
Additionally, measured data of blood glucose sensor was examined in order to detect adversarial
511
and accidental data modification intrusions (Verner & Butvinik 2017). Here, Otsu’s thresholding
512
algorithm was used with extra statistical analysis to create different informative feature vectors. 513
Then, linear SVM with different misclassification parameters were used to classify the feature
514
vectors. The obtained results on huge patient’s data showed 100% precision and 99.22% recall. 515
This demonstrated that data modification attacks can be effectively detected by utilizing the ML
516
approach. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed To assess the performance of the algorithm in
535
terms of accuracy and reliability, real and false observations were listed and estimated at the run
536
time. 529
Rathore et al. (Rathore et al. 2019) proposed a Long Short-Term Memory, which is a type of
530
recurrent neural network. It was used for predicting and forecasting different signal patterns of
531
Deep Brain Stimulation (DBS). Usually, Rest Tremor Velocity (RTV) is analyzed to know the
532
neurological disorders intensity. In this way, the authors analyzed RTV values for designing and
533
training the neural network. Multiple attacks have been introduced in the DBS context to
534
simulate and distinguish various attack techniques. To assess the performance of the algorithm in
535
terms of accuracy and reliability, real and false observations were listed and estimated at the run
536
time. Manuscript to be reviewed The
548
suggested methodology is the first effort to use ECG signals to exploit MLP in authentication. 549
The results were verified on the ECG dataset and showed that it is possible to achieve 100% test
550
accuracy with 5-degree coefficients when the authorized person is identified. 551
In a research made by Pirbhulal et al. (Pirbhulal et al. 2019), a biometric protection system based
552
on ML technique was introduced, in which the attributes for learning process were derived from
553
ECG signals. Nevertheless, in the testing process, the user is authenticated by using the unique
554
biometric EIs generated from the ECG and the polynomial approximation coefficients. It was
555
concluded that the proposed system can be used for real-time healthcare implementations. 517
Furthermore, other research groups (Kintzlinger et al. 2020) presented CardiWall system, which
518
is used for detecting and preventing resource depletion attacks against ICDs at programmer
519
device. The system is a collection of six security layers, exploitation of health experts
520
understanding, statistical techniques and one class SVM as the ML technique. To perform the
521
assessment, data were collected over a time span of four years and 775 benevolent clinical
522
commands were used. These were belonging to hundreds of specific patients and 28 malicious
523
clinical prompts established by two cardiac specialists. The evaluation results showed that only
524
two out of the six layers proposed in CardiWall system have provided a high detection
525
capability. One class SVM was failed to obtain a high detection rate because of the problems
526
related to the datasets and a smaller number of features related to the malicious data.. Noticeably,
527
the system achieved an AUC of 94.7% with a true positive rate (TPR) of 91.4% and false
528
positive rate (FPR) of 1%. 529
Rathore et al. (Rathore et al. 2019) proposed a Long Short-Term Memory, which is a type of
530
recurrent neural network. It was used for predicting and forecasting different signal patterns of
531
Deep Brain Stimulation (DBS). Usually, Rest Tremor Velocity (RTV) is analyzed to know the
532
neurological disorders intensity. In this way, the authors analyzed RTV values for designing and
533
training the neural network. Multiple attacks have been introduced in the DBS context to
534
simulate and distinguish various attack techniques. Manuscript to be reviewed 517
Furthermore, other research groups (Kintzlinger et al. 2020) presented CardiWall system, which
518
is used for detecting and preventing resource depletion attacks against ICDs at programmer
519
device. The system is a collection of six security layers, exploitation of health experts
520
understanding, statistical techniques and one class SVM as the ML technique. To perform the
521
assessment, data were collected over a time span of four years and 775 benevolent clinical
522
commands were used. These were belonging to hundreds of specific patients and 28 malicious
523
clinical prompts established by two cardiac specialists. The evaluation results showed that only
524
two out of the six layers proposed in CardiWall system have provided a high detection
525
capability. One class SVM was failed to obtain a high detection rate because of the problems
526
related to the datasets and a smaller number of features related to the malicious data.. Noticeably,
527
the system achieved an AUC of 94.7% with a true positive rate (TPR) of 91.4% and false
528
positive rate (FPR) of 1%. 529
Rathore et al. (Rathore et al. 2019) proposed a Long Short-Term Memory, which is a type of
530
recurrent neural network. It was used for predicting and forecasting different signal patterns of
531
Deep Brain Stimulation (DBS). Usually, Rest Tremor Velocity (RTV) is analyzed to know the
532
neurological disorders intensity. In this way, the authors analyzed RTV values for designing and
533
training the neural network. Multiple attacks have been introduced in the DBS context to
534
simulate and distinguish various attack techniques. To assess the performance of the algorithm in
535
terms of accuracy and reliability, real and false observations were listed and estimated at the run
536
time. 537
538
Table 8:
539
Details of published studies on anomaly and attack detection to the sensors/medical
540
devices. 541
542
Authentication and Access Control: In this section, results and analysis of the relevant papers
543
on this topic are given, while the main features and limitations of the reported works are
544
summarized in Table 9. 545
Rathore et al. (Rathore et al. 2018b) proposed a biometric verification methodology based on
546
ECG, in which the Legendre polynomial extraction and MLP classifier were used for
547
identification and authorization, aiming at securing the data, network, and application layer. 539
Details of published studies on anomaly and attack detection to the sensors/medical
540
devices.
541 542
Authentication and Access Control: In this section, results and analysis of the relevant papers
543
on this topic are given, while the main features and limitations of the reported works are
544
summarized in Table 9. 549
The results were verified on the ECG dataset and showed that it is possible to achieve 100% test
550
accuracy with 5-degree coefficients when the authorized person is identified. 551
In a research made by Pirbhulal et al. (Pirbhulal et al. 2019), a biometric protection system based
552
on ML technique was introduced, in which the attributes for learning process were derived from
553
ECG signals. Nevertheless, in the testing process, the user is authenticated by using the unique
554
biometric EIs generated from the ECG and the polynomial approximation coefficients. It was
555
concluded that the proposed system can be used for real-time healthcare implementations. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 556
In another work carried out by Mawgoud et al. (Mawgoud et al. 2019), an authentication
557
approach was proposed, which is based on SVM. The authentication was established by
558
incorporating both trust management and SVM at the gateway to recognize the frequency of
559
resource-constraint devices and the timing of access. The proposed approach acts to identify
560
several IoMT sensor artifacts based on their pseudo-random exposure in both frequency and time
561
domains. If the value of the time or frequency domain is the same as its distinctive Pseudo-
562
Random Binary Sequence (PRBS), the device will be authenticated by the gateway, in which an
563
average trust value of 1.9 and less than 0.5 value are considered as adversary. Results proved that
564
the method is viable for interacting with the IoMT devices. 558
incorporating both trust management and SVM at the gateway to recognize the frequency of
559
resource-constraint devices and the timing of access. The proposed approach acts to identify
560
several IoMT sensor artifacts based on their pseudo-random exposure in both frequency and time
561
domains. If the value of the time or frequency domain is the same as its distinctive Pseudo-
562
Random Binary Sequence (PRBS), the device will be authenticated by the gateway, in which an
563
average trust value of 1.9 and less than 0.5 value are considered as adversary. Results proved that
564
the method is viable for interacting with the IoMT devices. 565
Furthermore, a study was performed by Barros et al (Barros et al. 2019) aiming at reducing the
566
computational cost, in which the features were extracted from ECG signal by utilizing only
567
fiduciary points measured from the acquisition of the signal. A set of ML techniques such as
568
MLP, RF, SVM and NB were used for the classification purpose. The ECG signal of Stress
569
Recognition Database was employed and it was proved that the method can be used for
570
continuous authentication with lower complexity using less than 10 features. 565
Furthermore, a study was performed by Barros et al (Barros et al. 2019) aiming at reducing the
566
computational cost, in which the features were extracted from ECG signal by utilizing only
567
fiduciary points measured from the acquisition of the signal. A set of ML techniques such as
568
MLP, RF, SVM and NB were used for the classification purpose. Manuscript to be reviewed The ECG signal of Stress
569
Recognition Database was employed and it was proved that the method can be used for
570
continuous authentication with lower complexity using less than 10 features. 566
computational cost, in which the features were extracted from ECG signal by utilizing only
567
fiduciary points measured from the acquisition of the signal. A set of ML techniques such as
568
MLP, RF, SVM and NB were used for the classification purpose. The ECG signal of Stress
569
Recognition Database was employed and it was proved that the method can be used for
570
continuous authentication with lower complexity using less than 10 features. 571
Hospital systems with huge number of patients requires proper authentication method. For that
572
purpose, Zhang et al. (Zhang et al. 2018) proposed a hybrid technique that combines fiducial and
573
non-fiducial features for accuracy improvement for authentication in healthcare systems. Feature
574
selection of 2DPCA was used to enhance the accuracy of Linear Discriminant Analysis (LDA)
575
and improving authentication of smart healthcare along with MapReduce. The PQRST peaks
576
were selected as the main fiducial features. For the ECG user recognition, an incrementing
577
training of the LDA algorithm was considered, utilizing the exposed fiducial and non-fiducial
578
features. The authors reported that detection based on fiduciary feature and Fast Fourier
579
Transform has provided a poor precision. Additionally, they concerned that growing the number
580
of features would lead to additional computing effort. 571
Hospital systems with huge number of patients requires proper authentication method. For that
572
purpose, Zhang et al. (Zhang et al. 2018) proposed a hybrid technique that combines fiducial and
573
non-fiducial features for accuracy improvement for authentication in healthcare systems. Feature
574
selection of 2DPCA was used to enhance the accuracy of Linear Discriminant Analysis (LDA)
575
and improving authentication of smart healthcare along with MapReduce. The PQRST peaks
576
were selected as the main fiducial features. For the ECG user recognition, an incrementing
577
training of the LDA algorithm was considered, utilizing the exposed fiducial and non-fiducial
578
features. The authors reported that detection based on fiduciary feature and Fast Fourier
579
Transform has provided a poor precision. Additionally, they concerned that growing the number
580
of features would lead to additional computing effort. Manuscript to be reviewed 581
Referring back to the individual authentication for daily use of wearable devices, researchers
582
(Shang & Wu 2019) used photoplethysmography (PPG) signals inspired by hand movements for
583
user authentication on smartwatches. To protect from attackers, the software first senses the
584
beginning and stopping point of any new hand gesture and decides if an attacker is present. This
585
was realized by comparing the features derived from the observed hand movements with those
586
obtained from the regular user. The training samples of the usual consumer have been used to
587
create a local outlier factor (LOF) model. The LOF model has the feature of few parameter
588
modification which helps to easily create a new classifier for a new consumer. In addition, LOF
589
can identify anomalies efficiently. This approach resulted in an overall authentication accuracy
590
of 96:31% and an overall true rejection rate of at least 91:64 % against two forms of attacks. 591
Authors claimed that this software-based solution can be integrated into any smartwatch with
592
PPG sensors for multi-factor authentication. 593
On the other hand, Musale et al. (Musale et al. 2019) proposed a lightweight authentication
594
approach using gait biometric for authenticating users of commercial smart watches. This was
595
realized by extracting the statistical features and person’s actions-related features from the 593
On the other hand, Musale et al. (Musale et al. 2019) proposed a lightweight authentication
594
approach using gait biometric for authenticating users of commercial smart watches. This was
595
realized by extracting the statistical features and person’s actions-related features from the PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 596
collected data of sensors, thereby improving both accuracy and efficiency. For this purpose,
597
techniques such as filters with RF, K-NN and MLP classifiers were used. Results demonstrated
598
that the system could achieve a higher accuracy by having a reduced feature and fewer sensed
599
dataset. This in turn renders the theoretical architecture more realistic and easily accessible to
600
wearable IoT devices with minimal processing resources and energy efficiency. 596
collected data of sensors, thereby improving both accuracy and efficiency. For this purpose,
597
techniques such as filters with RF, K-NN and MLP classifiers were used. Results demonstrated
598
that the system could achieve a higher accuracy by having a reduced feature and fewer sensed
599
dataset. This in turn renders the theoretical architecture more realistic and easily accessible to
600
wearable IoT devices with minimal processing resources and energy efficiency. 601
Furthermore, in a study reported by (Vhaduri & Poellabauer 2019), a hybrid approach was
602
implemented for biometric authentication in wearable devices. This was achieved by utilizing a
603
three-combination form of course-grain minute-level biometrics: behavioral (step counts),
604
physiological (heart rate), and hybrid (calorie burn and metabolic equivalent of task). An
605
analysis was performed by using 400 Fitbit consumers data that were obtained from a health
606
research collected over 17 months. A combination of filters (KS-test, PC and SD) with SVM of
607
different settings were used. A high accuracy for both sedentary and non-sedentary with low
608
error rates of 0.05 were achieved when a binary SVM was used. It was also proved that
609
hybridizing the biometrics gives better results even during non-sedentary periods. 610
Mohsen et al. (Mohsen et al. 2019) proposed ECC defined lightweight cooperative
611
authentication scheme to be used in real-time medical wireless sensor networks among
612
physicians/nursing staff, trustworthy databases, sensors and patients. Here, the doctor/nurse can
613
use the system by his/her fingerprint, while patient identity can be verified through continuous
614
monitoring of physiological data (e.g., ECG signals) in every 30 mins to detect the physical theft
615
of the sensor. The server uses PID and ECC feature set as training data and utilizes convolutional
616
neural network. Dynamic identity was applied for user anonymity and mitigating user
617
traceability. Results showed that the scheme is strong against multiple attacks with accepted
618
performance 610
Mohsen et al. (Mohsen et al. Manuscript to be reviewed 636
technique of AES encryption, blockchain, and PSO steganography was used for secure
637
transmission of the data. In the evaluation process, 106 samples were chosen from a database
638
comprising of 6000 samples of FV images. It was seen that the system is strong against spoofing
639
and brute-force attacks, whereby an improvement of 55.56% was achieved in the secure
640
biometric transmission. 641
642
Table 9:
643
The summary of the studies reported on authentication and access control. 644
645
Intrusion and Malware Detection: Herein, the main achieved results on this topic that were
646
reported in literature are given followed by a summarized comparison of the studies, as shown in
647
Table 10. 648
Begli et al. (Begli et al. 2019) designed a security framework for smart healthcare system, which
649
can be specifically useful for protecting wireless sensor network against unauthorized access and
650
network attacks. In their work, a multiagent based layered architecture was first defined. Then,
651
the IDS was applied using SVM, which is proportional to the level of energy and the sensitivity
652
of the available data of each triple group of the agents. Different rules were used for each layer
653
of the multiagent healthcare. Also, attacks related to healthcare, including eavesdropping attacks,
654
were studied in terms of energy usage and computational cost. 655
Moreover, a multivariate correlation analysis was proposed for the IoMT attacks detection (Itten
656
& Vadakkumcheril 2016). Triangular based area maps were utilized for analyzing the incoming
657
data’s features to formulate a correlation among various features. It was seen that the learning
658
model has a high rate of time usage with increased classification accuracy. The proposed method
659
can differentiate both known and unknown DoS attacks from normal network traffic. 660
Additionally, in a study made by (Schneble & Thamilarasu 2019a), a massively distributed
661
intrusion detection based on ML was designed and implemented for the Medical Cyber Physical
662
System. Primarily, the notion of Federated Learning was researched to reduce the connectivity
663
and computing expenses associated with conventional ML algorithms. Evaluation was carried
664
out using real patient records against the attacks including DoS, data manipulation and false data
665
injection. Observational findings showed that the proposed system attained a high accuracy of
666
99% and FPR of 1% including a decreased communication network’s overhead. Manuscript to be reviewed Moreover, they
667
proved that the program could cope with unevenly distributed data and is a flexible approach that
668
utilizes the computational power of multiple mobile devices. In another work (Odesile &
669
Thamilarasu 2017), the authors have introduced a mobile agent-based intrusion detection
670
program for WBAN. Different types of sensor agents, and cluster agents are used . Multiple
671
threats were detected in a distributed and hierarchical framework within the healthcare network. 672
ML methods such as NBC, KNN, SVM, RF and DT were applied to sensor nodes for the
673
purpose of providing precise attack detection, followed by choosing an appropriate approach. 674
The system has been tested to be effective in terms of precision and power usage. It was
675
revealed that there is a lack of sufficient researches on the intrusion detection in the medical 636
technique of AES encryption, blockchai
637
transmission of the data. In the evaluatio
638
comprising of 6000 samples of FV imag
639
and brute-force attacks, whereby an imp
640
biometric transmission. 641
642
Table 9:
643
The summary of the studies reported
644
645
Intrusion and Malware Detection: He
646
reported in literature are given followed
647
Table 10. 648
Begli et al. (Begli et al. 2019) designed
649
can be specifically useful for protecting
650
network attacks. In their work, a multiag
651
the IDS was applied using SVM, which
652
of the available data of each triple group
653
of the multiagent healthcare. Also, attac
654
were studied in terms of energy usage an
655
Moreover, a multivariate correlation ana
656
& Vadakkumcheril 2016). Triangular ba
657
data’s features to formulate a correlation
658
model has a high rate of time usage with
659
can differentiate both known and unkno
660
Additionally, in a study made by (Schne
661
intrusion detection based on ML was de
662
System. Primarily, the notion of Federat
663
and computing expenses associated with
664
out using real patient records against the
665
injection. Observational findings showe
666
99% and FPR of 1% including a decreas
667
proved that the program could cope with
668
utilizes the computational power of mul
669
Thamilarasu 2017), the authors have int
670
program for WBAN. Manuscript to be reviewed 2019) proposed ECC defined lightweight cooperative
611
authentication scheme to be used in real-time medical wireless sensor networks among
612
physicians/nursing staff, trustworthy databases, sensors and patients. Here, the doctor/nurse can
613
use the system by his/her fingerprint, while patient identity can be verified through continuous
614
monitoring of physiological data (e.g., ECG signals) in every 30 mins to detect the physical theft
615
of the sensor. The server uses PID and ECC feature set as training data and utilizes convolutional
616
neural network. Dynamic identity was applied for user anonymity and mitigating user
617
traceability. Results showed that the scheme is strong against multiple attacks with accepted
618
performance. 619
Moreover, Punithavathi et al. (Punithavathi et al. 2019) proposed a lightweight approach for
620
cloud cancelable biometric authentication. As such, privacy problems related to biometric
621
utilization can be tackled by getting advantage of cancelable biometric templates. A random
622
projection method was used for generating the cancelable fingerprint template. Fingerprint
623
images were chosen anonymously from publicly accessible databases. After pre-processing and
624
feature extraction, the lightweight random projection method was used to create cancelable
625
fingerprint models. However, authors mentioned that the work’s scalability in real world has not
626
been tested. 627
Rathore et al. (Rathore et al. 2018a) implemented a lightweight approach for trusted
628
authentication using Dynamic Time Warping (DTW) method. A large database from Physionet
629
was used for validating the method and informative features were extracted from ECG using
630
DTW. It was seen that DTW method showed a higher accuracy compared with that of the non-
631
linear SVM. Additionally, it took less time, while both methods have the same memory
632
complexity. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed Different types of
671
threats were detected in a distributed an
672
ML methods such as NBC, KNN, SVM
673
purpose of providing precise attack dete
674
The system has been tested to be effecti
675
revealed that there is a lack of sufficient 636
technique of AES encryption, blockchain, and PSO steganography was used for secure
637
transmission of the data. In the evaluation process, 106 samples were chosen from a database
638
comprising of 6000 samples of FV images. It was seen that the system is strong against spoofing
639
and brute-force attacks, whereby an improvement of 55.56% was achieved in the secure
640
biometric transmission. 641
642
Table 9:
643
The summary of the studies reported on authentication and access control. 644
645
Intrusion and Malware Detection: Herein, the main achieved results on this topic that were
646
reported in literature are given followed by a summarized comparison of the studies, as shown in
647
Table 10. 648
Begli et al. (Begli et al. 2019) designed a security framework for smart healthcare system, which
649
can be specifically useful for protecting wireless sensor network against unauthorized access and
650
network attacks. In their work, a multiagent based layered architecture was first defined. Then,
651
the IDS was applied using SVM, which is proportional to the level of energy and the sensitivity
652
of the available data of each triple group of the agents. Different rules were used for each layer
653
of the multiagent healthcare. Also, attacks related to healthcare, including eavesdropping attacks,
654
were studied in terms of energy usage and computational cost. 655
Moreover, a multivariate correlation analysis was proposed for the IoMT attacks detection (Itten
656
& Vadakkumcheril 2016). Triangular based area maps were utilized for analyzing the incoming
657
data’s features to formulate a correlation among various features. It was seen that the learning 636
technique of AES encryption, blockchain, and PSO steganography was used for secure
637
transmission of the data. In the evaluation process, 106 samples were chosen from a database
638
comprising of 6000 samples of FV images. It was seen that the system is strong against spoofing
639
and brute-force attacks, whereby an improvement of 55.56% was achieved in the secure
640
biometric transmission. The summary of the studies reported on authentication and access control. 655
Moreover, a multivariate correlation analysis was proposed for the IoMT attacks detection (Itten
656
& Vadakkumcheril 2016). Triangular based area maps were utilized for analyzing the incoming
657
data’s features to formulate a correlation among various features. It was seen that the learning
658
model has a high rate of time usage with increased classification accuracy. The proposed method
659
can differentiate both known and unknown DoS attacks from normal network traffic. 655
Moreover, a multivariate correlation analysis was proposed for the IoMT attacks detection (Itten
656
& Vadakkumcheril 2016). Triangular based area maps were utilized for analyzing the incoming
657
data’s features to formulate a correlation among various features. It was seen that the learning
658
model has a high rate of time usage with increased classification accuracy. The proposed method
659
can differentiate both known and unknown DoS attacks from normal network traffic. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed The measurement criteria used
704
in the study were precision, recall, accuracy, and F1-score. The suggested model achieved
705
positive results across all variables compared to the other techniques. It was claimed that this
706
model can be expanded for detecting several forms of attacks against IoT devices and different
707
databases. 708
Alrashdi et al. (Alrashdi et al. 2019) presented a fog-based attack detection (FBAD) architecture
709
by utilizing an online sequential extreme learning machine (EOS-ELM) collection for
710
monitoring of suspicious behaviors in healthcare system. They proved that the proposed
711
architecture is effectively implemented in the decentralized fog-attack detection by comparing its
712
efficiency to other methods. It was revealed that the decentralized architecture surpassed the 676
cyber-physical system (Schneble & Thamilarasu 2019b). Since real time attack detection is
677
critical for medical devices, the volume of data analyzed by the IDS should be reduced to
678
maintain optimal frequency detection. Therefore, an effective IDS was proposed by addressing
679
the problem of feature selection to reduce the data dimensionality (Schneble & Thamilarasu
680
2019b). The findings showed that Laplacian scoring strategies are effective in optimizing the
681
collection of features with reduced resource usage. 682
Furthermore, for the detection of anomaly based intrusion, a stacked autoencoder (SAE) was
683
proposed by He et al. (He et al. 2019). The SAE was used to extract more informative features
684
and eliminate feature dimensions, which resulted in the reduction of detection overhead. 685
RM et al (RM et al. 2020) applied a hybrid PCA-GWO algorithm for feature selection and DNN
686
classifier for classifying the network attacks. The proposed methodology suits the IoMT devices
687
that are using a unique IP. One-Hot encoding scheme was used for pre-processing the input data. 688
Then, PCA and GWO algorithms were sequentially utilized for further data reduction followed
689
by the use of well-known classifiers for prediction. Results showed that the hybrid PCA-GWO is
690
capable of increasing the detection accuracy by 15%. In addition, the training and classification
691
time was reduced by 32%. 676
cyber-physical system (Schneble & Thamilarasu 2019b). Since real time attack detection is
677
critical for medical devices, the volume of data analyzed by the IDS should be reduced to
678
maintain optimal frequency detection. Manuscript to be reviewed Therefore, an effective IDS was proposed by addressing
679
the problem of feature selection to reduce the data dimensionality (Schneble & Thamilarasu
680
2019b). The findings showed that Laplacian scoring strategies are effective in optimizing the
681
collection of features with reduced resource usage. 685
RM et al (RM et al. 2020) applied a hybrid PCA-GWO algorithm for feature selection and DNN
686
classifier for classifying the network attacks. The proposed methodology suits the IoMT devices
687
that are using a unique IP. One-Hot encoding scheme was used for pre-processing the input data. 688
Then, PCA and GWO algorithms were sequentially utilized for further data reduction followed
689
by the use of well-known classifiers for prediction. Results showed that the hybrid PCA-GWO is
690
capable of increasing the detection accuracy by 15%. In addition, the training and classification
691
time was reduced by 32%. 692
For the same purpose, researchers (Thamilarasu 2016) used a multi-objective GA algorithm for
693
the feature selection of WBAN network attack detection. The experimental results showed that
694
the proposed algorithm could include the useful features for detecting a specific attack in the
695
detection process, thereby decreasing the computational complexity. 696
Asfaw et al. (Asfaw et al. 2010) presented a datamining-based model to provide a host-based
697
anomaly and attack detection method for pervasive healthcare systems. When a mobile is
698
requesting from the server, its message is recorded and fed to the classification model. Then, the
699
model classifies the record as benign or malicious depending on the previously recorded history. 700
Eventually, the classifier holds the reactions in a passive fashion, maintaining each specific
701
record with the disruptive behavior and hence detecting the anomalous events. 702
In a recent work performed by Manimurugan et al. (Manimurugan et al. 2020), Deep Belief
703
Network (DBN) was proposed for attacks detection in the IoMT. The measurement criteria used
704
in the study were precision, recall, accuracy, and F1-score. The suggested model achieved
705
positive results across all variables compared to the other techniques. It was claimed that this
706
model can be expanded for detecting several forms of attacks against IoT devices and different
707
databases. 708
Alrashdi et al. (Alrashdi et al. 2019) presented a fog-based attack detection (FBAD) architecture
709
by utilizing an online sequential extreme learning machine (EOS-ELM) collection for
710
monitoring of suspicious behaviors in healthcare system. Manuscript to be reviewed 676
cyber-physical system (Schneble & Thamilarasu 2019b). Since real time attack detection is
677
critical for medical devices, the volume of data analyzed by the IDS should be reduced to
678
maintain optimal frequency detection. Therefore, an effective IDS was proposed by addressing
679
the problem of feature selection to reduce the data dimensionality (Schneble & Thamilarasu
680
2019b). The findings showed that Laplacian scoring strategies are effective in optimizing the
681
collection of features with reduced resource usage. 682
Furthermore, for the detection of anomaly based intrusion, a stacked autoencoder (SAE) was
683
proposed by He et al. (He et al. 2019). The SAE was used to extract more informative features
684
and eliminate feature dimensions, which resulted in the reduction of detection overhead. 685
RM et al (RM et al. 2020) applied a hybrid PCA-GWO algorithm for feature selection and DNN
686
classifier for classifying the network attacks. The proposed methodology suits the IoMT devices
687
that are using a unique IP. One-Hot encoding scheme was used for pre-processing the input data. 688
Then, PCA and GWO algorithms were sequentially utilized for further data reduction followed
689
by the use of well-known classifiers for prediction. Results showed that the hybrid PCA-GWO is
690
capable of increasing the detection accuracy by 15%. In addition, the training and classification
691
time was reduced by 32%. 692
For the same purpose, researchers (Thamilarasu 2016) used a multi-objective GA algorithm for
693
the feature selection of WBAN network attack detection. The experimental results showed that
694
the proposed algorithm could include the useful features for detecting a specific attack in the
695
detection process, thereby decreasing the computational complexity. 696
Asfaw et al. (Asfaw et al. 2010) presented a datamining-based model to provide a host-based
697
anomaly and attack detection method for pervasive healthcare systems. When a mobile is
698
requesting from the server, its message is recorded and fed to the classification model. Then, the
699
model classifies the record as benign or malicious depending on the previously recorded history. 700
Eventually, the classifier holds the reactions in a passive fashion, maintaining each specific
701
record with the disruptive behavior and hence detecting the anomalous events. 702
In a recent work performed by Manimurugan et al. (Manimurugan et al. 2020), Deep Belief
703
Network (DBN) was proposed for attacks detection in the IoMT. Manuscript to be reviewed They proved that the proposed
711
architecture is effectively implemented in the decentralized fog-attack detection by comparing its
712
efficiency to other methods. It was revealed that the decentralized architecture surpassed the
713
centralized framework in terms of detection time and accuracy of classification. 708
Alrashdi et al. (Alrashdi et al. 2019) presented a fog-based attack detection (FBAD) architecture
709
by utilizing an online sequential extreme learning machine (EOS-ELM) collection for
710
monitoring of suspicious behaviors in healthcare system. They proved that the proposed
711
architecture is effectively implemented in the decentralized fog-attack detection by comparing its
712
efficiency to other methods. It was revealed that the decentralized architecture surpassed the
713
centralized framework in terms of detection time and accuracy of classification. 714
Wazid et al. (Wazid et al. 2019) surveyed the malware detection methods in the IoMT network
715
using ML approaches. They elaborated on how serious the malware attacks are, especially botnet 714
Wazid et al. (Wazid et al. 2019) surveyed the malware detection methods in the IoMT network
715
using ML approaches. They elaborated on how serious the malware attacks are, especially botnet PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed Their study concluded that
744
breaching those identical brain activities are possible if proper security measures are not taken
745
into consideration. 746
747
748
Table 10:
749
Summary of the studies reported on intrusion and malware detection. 750
751
752
Discussion
753
In this section, the main outcomes of the reviewed studies are elaborated and discussed aiming at
754
achieving fruitful answers to each of the research questions, respectively. 716
attacks on the three tiers of security and privacy. In the presence of such attacks, the sensitive
717
data of IoT communication may be disclosed, altered or even may not be available to the
718
authorized users. Hence, in the study various types of malware attacks were explored with their
719
symptoms and a taxonomy for the IoMT security was given. Moreover, ML based malware
720
detection methods were discussed. 721
Furthermore, Fernandez Maimo et al.(Fernandez Maimo et al. 2019) used ML techniques for
722
detecting and classifying ransomware attacks in ICE. The NFV/SDN methods were used to
723
isolate and remove contaminated medical equipment and networks. The method was developed
724
to detect recent malwares such as WannaCry, Petya, BadRabbit and PowerGhost. Techniques
725
such as OC-SVM and Naive Bayes have been proved to detect and classify ransomware infecting
726
ICE with respective accuracy and recall of 92.32% and 99.97% for the OC-SVM in anomaly
727
detection. This is where the Naive Bayes classifier was able to reach a classification accuracy of
728
99.99%. 729
Shakeel et al. (Shakeel et al. 2018) studied a secure data access and transmission in the IoMT
730
through utilizing the Deep-Q-Network (DQN) methodology. Initially, the IoMT system was
731
analyzed by the deep neural network to authenticate and eliminate any possible malware attacks. 732
The traffic attributes of each request were derived and recorded in the database. The output
733
attribute was then analyzed by using the state feature and associated behavior. Then, the deep
734
neural convolution network was used to classify them into malware and regular data. Results
735
showed that the method has achieved a low error of about 0.12 with malware detection rate of
736
98.79%. 729
Shakeel et al. (Shakeel et al. 2018) studied a secure data access and transmission in the IoMT
730
through utilizing the Deep-Q-Network (DQN) methodology. Manuscript to be reviewed 716
attacks on the three tiers of security and privacy. In the presence of such attacks, the sensitive
717
data of IoT communication may be disclosed, altered or even may not be available to the
718
authorized users. Hence, in the study various types of malware attacks were explored with their
719
symptoms and a taxonomy for the IoMT security was given. Moreover, ML based malware
720
detection methods were discussed. 721
Furthermore, Fernandez Maimo et al.(Fernandez Maimo et al. 2019) used ML techniques for
722
detecting and classifying ransomware attacks in ICE. The NFV/SDN methods were used to
723
isolate and remove contaminated medical equipment and networks. The method was developed
724
to detect recent malwares such as WannaCry, Petya, BadRabbit and PowerGhost. Techniques
725
such as OC-SVM and Naive Bayes have been proved to detect and classify ransomware infecting
726
ICE with respective accuracy and recall of 92.32% and 99.97% for the OC-SVM in anomaly
727
detection. This is where the Naive Bayes classifier was able to reach a classification accuracy of
728
99.99%. 729
Shakeel et al. (Shakeel et al. 2018) studied a secure data access and transmission in the IoMT
730
through utilizing the Deep-Q-Network (DQN) methodology. Initially, the IoMT system was
731
analyzed by the deep neural network to authenticate and eliminate any possible malware attacks. 732
The traffic attributes of each request were derived and recorded in the database. The output
733
attribute was then analyzed by using the state feature and associated behavior. Then, the deep
734
neural convolution network was used to classify them into malware and regular data. Results
735
showed that the method has achieved a low error of about 0.12 with malware detection rate of
736
98.79%. 737
Furthermore, with the aim of protecting the brain activity data from attacks in Brain–Computer
738
Interface (BCI) systems such as Electroencephalography (EEG) and fraud BCI program (e.g. 739
game), which lets a malicious entity to obtain the user’s brain activities, researchers
740
demonstrated that a ML method can anticipate relevant identity characteristics by studying
741
resting-state EEG (rsEEG) records of a person's brain activity (Landau et al. 2020). A complete
742
collection of raw rsEEG tests along with dissociation degree and executive function (EF)
743
assessment indicators were used for 162 subjects in the test. 775
RQ 2: What kind of data and tools are used for applying ML techniques in
th
I MT
it ? y
777
Machine Learning methods are data dependent as they learn from these data overtime and decide
778
intelligently based on their learning ability, amount and quality of the data. For this reason, in
779
our study we have paid attention to this matter and we have analyzed all the selected research
780
articles based on the type of data they used for decision making and learning process. It can be
781
seen from Fig. 8 that most of the papers have used historical benchmark data, of which 9% have
782
used network data, while 51% have used sensors and physiological data. This is mainly because
783
most of the methods were to find anomalies in the sensors and to use device authentication as
784
security solution.. On the other hand, 26% of the papers have used simulated or emulated data. 785
However, some of the studies have not given the source of their data or did not mention it at all. 786
For this category, we have given the label not available or not given. This group of papers
787
provides 14% of the whole selected papers. Manuscript to be reviewed Initially, the IoMT system was
731
analyzed by the deep neural network to authenticate and eliminate any possible malware attacks. 732
The traffic attributes of each request were derived and recorded in the database. The output
733
attribute was then analyzed by using the state feature and associated behavior. Then, the deep
734
neural convolution network was used to classify them into malware and regular data. Results
735
showed that the method has achieved a low error of about 0.12 with malware detection rate of
736
98.79%. 735
showed that the method has achieved a low error of about 0.12 with malware detection rate of
736
98.79%. 737
Furthermore, with the aim of protecting the brain activity data from attacks in Brain–Computer
738
Interface (BCI) systems such as Electroencephalography (EEG) and fraud BCI program (e.g. 739
game), which lets a malicious entity to obtain the user’s brain activities, researchers
740
demonstrated that a ML method can anticipate relevant identity characteristics by studying
741
resting-state EEG (rsEEG) records of a person's brain activity (Landau et al. 2020). A complete
742
collection of raw rsEEG tests along with dissociation degree and executive function (EF)
743
assessment indicators were used for 162 subjects in the test. Their study concluded that
744
breaching those identical brain activities are possible if proper security measures are not taken
745
into consideration. 746
747
748
Table 10:
749
Summary of the studies reported on intrusion and malware detection. 750
751
752
Discussion
753
In this section, the main outcomes of the reviewed studies are elaborated and discussed aiming at
754
achieving fruitful answers to each of the research questions, respectively. 753
In this section, the main outcomes of the reviewed studies are elaborated and discussed aiming at
754
achieving fruitful answers to each of the research questions, respectively. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Figure 7: Papers distribution by the direction and problem-solving domain. Figure 7: Papers distribution by the direction and problem-solving domain. Manuscript to be reviewed 755
756
RQ1: What is the current state of the art and direction of study in the IoMT
757
security using ML? 758
One can see from Fig. 7 that 37% of the research articles were devoted to detect anomaly at
759
sensors or medical devices. This indicated that in the past years, the main focus was on detecting
760
the intrusions such as false data injection, resource depletion attacks, behavioral and physical
761
attacks against medical devices. This can be attributed to the fact that these attacks are serious
762
and they lead to significant health issues. For instance, it may lead to death in the case of attacks
763
to the implantable medical devices (Alsubaei et al. 2019b; Hatzivasilis et al. 2019; Yaacoub et al. 764
2020). Computer Science Computer Science 756
RQ1: What is the current state of the art and direction of study in the IoMT
757
security using ML? 756
RQ1: What is the current state of the art and direction of study in the IoMT
757
security using ML? 758
One can see from Fig. 7 that 37% of the research articles were devoted to detect anomaly at
759
sensors or medical devices. This indicated that in the past years, the main focus was on detecting
760
the intrusions such as false data injection, resource depletion attacks, behavioral and physical
761
attacks against medical devices. This can be attributed to the fact that these attacks are serious
762
and they lead to significant health issues. For instance, it may lead to death in the case of attacks
763
to the implantable medical devices (Alsubaei et al. 2019b; Hatzivasilis et al. 2019; Yaacoub et al. 764
2020). 765
Also, intrusion and malware attack detections received a considerable attention, counting for
766
35% of the reported studies (7% for malware and 28% for other attacks). Nevertheless, majority
767
of the papers were found to have focused on NIDS. On the other hand, papers that used different
768
strategies to solve the authentication and access control issues were found to take 28%. However,
769
as authentication is computationally heavy for the IoMT devices, the current direction of
770
research is to apply lightweight mechanisms (Aghili et al. 2019a; Wu et al. 2018) by using sensor
771
physiological data to reduce the computation load on the device. 772
773
Figure 7: Papers distribution by the direction and problem-solving domain. 774
775
RQ 2: What kind of data and tools are used for applying ML techniques in
776
the IoMT security? 777
Machine Learning methods are data dependent as they learn from these data overtime and decide
778
intelligently based on their learning ability, amount and quality of the data. For this reason, in
779
our study we have paid attention to this matter and we have analyzed all the selected research
780
articles based on the type of data they used for decision making and learning process. It can be
781
seen from Fig. 8 that most of the papers have used historical benchmark data, of which 9% have
782
used network data, while 51% have used sensors and physiological data. This is mainly because
783
most of the methods were to find anomalies in the sensors and to use device authentication as
784
security solution.. 756
RQ1: What is the current state of the art and direction of study in the IoMT
757
security using ML? On the other hand, 26% of the papers have used simulated or emulated data. 785
However, some of the studies have not given the source of their data or did not mention it at all. 786
For this category, we have given the label not available or not given. This group of papers
787
provides 14% of the whole selected papers. 788
789
Figure 8. The type of data used by the researchers to conduct their research. 790
791
To further answer the above research question, software and hardware tools used and reported in
792
the reviewed studies are analyzed. Figure 9 shows the used software tools and programs in the
793
reviewed studies. Tools used by the studied works were mainly Network/ Sensor Simulators with
794
ML tools. However, we have excluded those studies that used simulation but did not mention its 765
Also, intrusion and malware attack detections received a considerable attention, counting for
766
35% of the reported studies (7% for malware and 28% for other attacks). Nevertheless, majority
767
of the papers were found to have focused on NIDS. On the other hand, papers that used different
768
strategies to solve the authentication and access control issues were found to take 28%. However,
769
as authentication is computationally heavy for the IoMT devices, the current direction of
770
research is to apply lightweight mechanisms (Aghili et al. 2019a; Wu et al. 2018) by using sensor
771
physiological data to reduce the computation load on the device. 770
research is to apply lightweight mechanisms (Aghili et al. 2019a; Wu et al. 2018) by using sensor
771
physiological data to reduce the computation load on the device. 789
Figure 8. The type of data used by the researchers to conduct their research. 790
791
To further answer the above research question, software and hardware tools used and reported in
792
the reviewed studies are analyzed. Figure 9 shows the used software tools and programs in the
793
reviewed studies. Tools used by the studied works were mainly Network/ Sensor Simulators with
794
ML tools. However, we have excluded those studies that used simulation but did not mention its 791
To further answer the above research question, software and hardware tools used and reported in
792
the reviewed studies are analyzed. Figure 9 shows the used software tools and programs in the
793
reviewed studies. Tools used by the studied works were mainly Network/ Sensor Simulators with 793
reviewed studies. Tools used by the studied works were mainly Network/ Sensor Simulators with
794
ML tools. However, we have excluded those studies that used simulation but did not mention its PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 795
tool. For this reason, the percentage of the simulation tools is less than expected. Furthermore,
796
25% of the studies did not give the tools used in their studies. The remaining were mainly
797
programming tools for ML purpose. We can see that MATLAB has been used more than the rest
798
of the tools. Python also gained good attention in the current years. In addition, Weka tool has
799
been used frequently, which counts for 9% of the studies. Additionally, Keras and Sckit-learn
800
libraries were used with 8% and 6% by the studies, respectively. Those represent open-source
801
libraries are usually used with Python. Moreover, in the reviewed studies, some works have used
802
testbeds and hardware tools, while few of them reported their tools. Figure 10 shows the number
803
of those tools that have been used. It was found that six types of devices and hardware tools were
804
used and reported in the studies. 805
806
Figure 9: Software and tools used by the studies. 807
Figure 10: Devices and hardware tools used in the studies. 808
809
810
RQ 3: How ML techniques are effectively applied by the studies and what
811
are their limitations ? 812
Through a critical analysis of the current works, we can conclude that traditional ML techniques
813
may fail if proper considerations are not given to some metrics such as computational complexity
814
and energy usage. ML performance is reduced when few data are used. However, the IoMT
815
devices are resource limited such as (IMDs) and using a huge data analysis on these devices is
816
resulted in degrading their lifespan. For these reasons, one should take a balance between these
817
two aspects. From our analysis of the reviewed papers, we have found that a small portion of the
818
studies have used lightweight approaches. The pie chart shown in Fig. 11-a illustrates that only
819
14% of the studies have used lightweight approach. Additionally, 12% of the studies have used
820
low complexity approaches, that means it is not exactly lightweight but not heavy. The rest of the
821
studies either did not require this feature in their system or failed to apply it. Furthermore, real
822
time ML techniques are crucial for the IoMT, especially in attack and intrusion detection. 810
RQ 3: How ML techniques are effectively applied by the studies and what
811
are their limitations ? 812
Through a critical analysis of the current works, we can conclude that traditional ML techniques
813
may fail if proper considerations are not given to some metrics such as computational complexity
814
and energy usage. ML performance is reduced when few data are used. However, the IoMT
815
devices are resource limited such as (IMDs) and using a huge data analysis on these devices is
816
resulted in degrading their lifespan. For these reasons, one should take a balance between these
817
two aspects. From our analysis of the reviewed papers, we have found that a small portion of the
818
studies have used lightweight approaches. The pie chart shown in Fig. 11-a illustrates that only
819
14% of the studies have used lightweight approach. Additionally, 12% of the studies have used
820
low complexity approaches, that means it is not exactly lightweight but not heavy. The rest of the
821
studies either did not require this feature in their system or failed to apply it. Furthermore, real
822
time ML techniques are crucial for the IoMT, especially in attack and intrusion detection. Manuscript to be reviewed It was foun
804
used and reported in the studies. 805
806
Figure 9: Software and tools used by the stu
807
Figure 10: Devices and hardware tools used
808
809
810
RQ 3: How ML techniques are effect
811
are their limitations ? 812
Through a critical analysis of the current work
813
may fail if proper considerations are not given
814
and energy usage. ML performance is reduced
815
devices are resource limited such as (IMDs) an
816
resulted in degrading their lifespan. For these
817
two aspects. From our analysis of the reviewed
818
studies have used lightweight approaches. The
819
14% of the studies have used lightweight appr
820
low complexity approaches, that means it is no
821
studies either did not require this feature in the
822
time ML techniques are crucial for the IoMT,
823
Because the IoMT networks are dealing with s
824
from working, a continuous and fast detection
825
that 19% of the studies were used real time ap
826
complex approaches that are not exactly real ti
827
67% of the works were offline. Additionally, t
828
paid less attention to distributed attack detectio
829
studied have used decentralized models, while
830
the methods was considered as a better approa
831
832
Figure 11: Analysis of the studies in terms o
833 Manuscript to be reviewed 823
Because the IoMT networks are dealing with streaming data and some of the devices never stop
824
from working, a continuous and fast detection of attacks are required. It is seen from Fig. 11-b
825
that 19% of the studies were used real time approach, while 12% of the studies have used low
826
complex approaches that are not exactly real time but have approached it. Nevertheless, the other
827
67% of the works were offline. Additionally, the studies focused on network attack detection
828
paid less attention to distributed attack detection. As can be seen in Fig. 12 that only 12% of the
829
studied have used decentralized models, while the rest were centralized. However, a hybrid of
830
the methods was considered as a better approach. 831
832
Figure 11: Analysis of the studies in terms of (a) complexity and (b) real time analysis
833 795
tool. For this reason, the percentage of the simulation tools is less than expected. Furthermore,
796
25% of the studies did not give the tools used in their studies. The remaining were mainly
797
programming tools for ML purpose. We can see that MATLAB has been used more than the rest
798
of the tools. Python also gained good attention in the current years. In addition, Weka tool has
799
been used frequently, which counts for 9% of the studies. Additionally, Keras and Sckit-learn
800
libraries were used with 8% and 6% by the studies, respectively. Those represent open-source
801
libraries are usually used with Python. Moreover, in the reviewed studies, some works have used
802
testbeds and hardware tools, while few of them reported their tools. Figure 10 shows the number
803
of those tools that have been used. It was found that six types of devices and hardware tools were
804
used and reported in the studies. 795
tool. For this reason, the percentage of the sim
796
25% of the studies did not give the tools used
797
programming tools for ML purpose. We can s
798
of the tools. Python also gained good attention
799
been used frequently, which counts for 9% of
800
libraries were used with 8% and 6% by the stu
801
libraries are usually used with Python. Moreov
802
testbeds and hardware tools, while few of them
803
of those tools that have been used. 833
834
Figure 12: Details of the studies in terms of placement.
835 PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 836
837
Based on the studies gaps that were assigned previously, it is found that majority of the studies
838
have focused on improving the traditional evaluation metrics for their ML models such as high
839
(accuracy, recall, precision) and low (FP, FN). However, performance metrics such as memory
840
(space), CPU, time, and energy overhead were neglected. Additionally, most of the techniques of
841
attack detection were performed in an isolated offline environment. However, one should
842
consider simulation/hardware implementation to represent a real IoT setting. Also, decentralized
843
and hybrid approaches are much compatible for the IoMT than centralized one, which was again
844
not common among the studies. Furthermore, we have noticed that the datasets used by the
845
attack and intrusion detection methods were out of date and some of them do not represent the
846
current IoMT system. Moreover, sensitive, and private data usage should be avoided in attack
847
and anomaly detection tasks which was also given a minor attention. Therefore, there should be a
848
trade-off among preserving privacy, high accuracy, and computational complexity. Another
849
challenge that should be considered is the risk of adversarial attacks on ML techniques
850
themselves. An attacker who have enough knowledge on how the ML techniques work with the
851
data can manipulate the data at training or testing stage to fool the ML method. Hence, the
852
proposed methods should be strong enough against the adversarial attacks. 853
854
855
Limitations
856
The underlying drawbacks of this research are: i) Selection bias: This study focuses on literature
857
research, which may have unintentionally omitted most recent non-scholarly advancements or
858
scholarly articles that yet to be published at the time of preparing this study. ii) Publication bias:
859
Some of the studies may have been discarded since their full paper was not accessible. iii)
860
Descriptive bias: Despite the efforts made to present the background comprehensively, this SLR
861
is combining many topics that may need someone to refer to the external sources for a deep
862
understanding. 863
864
Conclusions
865
In this study, a comprehensive Systematic Literature Review (SLT) was given about the IoMT
866
security and privacy issues and how Machine Learning (ML) methods are used for solving them. 867
By examining the content of the study, including methods, good features, limitations, tools, and
868
datasets, the designated research questions were answered. 888
References 889
Abdaoui A, Al-Ali A, Riahi A, Mohamed A, Du X, and Guizani M. 2020. Secure medical treatment with
890
deep learning on embedded board. Energy Efficiency of Medical Devices and Healthcare
891
Applications: Elsevier, 131-151. 892
Abhishek NV, Lim TJ, Sikdar B, and Tandon A. 2018. An intrusion detection system for detecting
893
compromised gateways in clustered IoT networks. 2018 IEEE International Workshop Technical
894
Committee on Communications Quality and Reliability (CQR): IEEE. p 1-6. 895
Aghili SF, Mala H, Kaliyar P, and Conti M. 2019a. SecLAP: Secure and lightweight RFID authentication
896
protocol for Medical IoT. Future Generation Computer Systems 101:621-634. 895
Aghili SF, Mala H, Kaliyar P, and Conti M. 2019a. SecLAP: Secure and lightweight RFID authentication
896
protocol for Medical IoT. Future Generation Computer Systems 101:621-634. 895
Aghili SF, Mala H, Kaliyar P, and Conti M. 2019a. SecLAP: Secure and lightweight RFID authentication
896
protocol for Medical IoT. Future Generation Computer Systems 101:621-634. 898
Aghili SF, Mala H, Shojafar M, and Peris-Lopez P. 2019b. LACO: Lightweight Three-Factor Authentication,
899
Access Control and Ownership Transfer Scheme for E-Health Systems in IoT. Future Generation
900
Computer Systems 96:410-424. https://doi.org/10.1016/j.future.2019.02.020 898
Aghili SF, Mala H, Shojafar M, and Peris-Lopez P. 2019b. LACO: Lightweight Three-Factor Authentication,
899
Access Control and Ownership Transfer Scheme for E-Health Systems in IoT. Future Generation
900
Computer Systems 96:410-424. https://doi.org/10.1016/j.future.2019.02.020 901
Ahad A, Tahir M, and Yau KA. 2019. 5G-Based Smart Healthcare Network: Architecture, Taxonomy,
902
Challenges and Future Research Directions. IEEE Access 7:100747-100762. 903
10.1109/ACCESS.2019.2930628 901
Ahad A, Tahir M, and Yau KA. 2019. 5G-Based Smart Healthcare Network: Architecture, Taxonomy,
902
Challenges and Future Research Directions. IEEE Access 7:100747-100762. 903
10.1109/ACCESS.2019.2930628 904
Ahmad B, Jian W, Ali ZA, Tanvir S, and Khan MSA. 2019. Hybrid anomaly detection by using clustering for
905
wireless sensor network. Wireless Personal Communications 106:1841-1853. 904
Ahmad B, Jian W, Ali ZA, Tanvir S, and Khan MSA. 2019. Hybrid anomaly detection by using clustering for
905
wireless sensor network. Wireless Personal Communications 106:1841-1853. 906
Al Shorman A, Faris H, and Aljarah I. 2019. Unsupervised intelligent system based on one class support
907
vector machine and Grey Wolf optimization for IoT botnet detection. Journal of Ambient
908
Intelligence and Humanized Computing:1-17. 906
Al Shorman A, Faris H, and Aljarah I. 2019. Unsupervised intelligent system based on one class support
907
vector machine and Grey Wolf optimization for IoT botnet detection. Manuscript to be reviewed Findings of this study showed that
869
ML techniques are effective in addressing the IoMT security issues with promising results. 870
Majority of the studies was devoted to device layer or body area network security since attacks
871
on devices such as IMDs are seriously affecting the patient’s health and life. The security
872
solutions for such devices were sensor anomaly detection and device authentication and access
873
control. Furthermore, securing the network layer was seen among the studies that used attack and
874
malware detection strategies. 836
837
Based on the studies gaps that were assigned previously, it is found that majority of the studies
838
have focused on improving the traditional evaluation metrics for their ML models such as high
839
(accuracy, recall, precision) and low (FP, FN). However, performance metrics such as memory
840
(space), CPU, time, and energy overhead were neglected. Additionally, most of the techniques of
841
attack detection were performed in an isolated offline environment. However, one should
842
consider simulation/hardware implementation to represent a real IoT setting. Also, decentralized
843
and hybrid approaches are much compatible for the IoMT than centralized one, which was again
844
not common among the studies. Furthermore, we have noticed that the datasets used by the
845
attack and intrusion detection methods were out of date and some of them do not represent the
846
current IoMT system. Moreover, sensitive, and private data usage should be avoided in attack
847
and anomaly detection tasks which was also given a minor attention. Therefore, there should be a
848
trade-off among preserving privacy, high accuracy, and computational complexity. Another
849
challenge that should be considered is the risk of adversarial attacks on ML techniques
850
themselves. An attacker who have enough knowledge on how the ML techniques work with the
851
data can manipulate the data at training or testing stage to fool the ML method. Hence, the
852
proposed methods should be strong enough against the adversarial attacks. 853 PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 875
Moreover, the tools and environment of the current works are a combination of network
876
simulators and ML tools with more focus on the latter. Additionally, there is a lack of relevant
877
datasets, especially in the intrusion detection. Most of the studies focused on improving the
878
common ML algorithms evaluation metrics such as high accuracy and low FPR. However, since
879
the IoMT devices are characterized with shortage in power and small memory and processor,
880
there should be a balance between security and maintaining resources lifespan during the
881
adaption of these solutions. We have concluded that traditional ML techniques may fail if proper
882
consideration is not given to some metrics such as resource complexity, time complexity, and
883
energy usage. It was noticed that a vast majority of the studies ignored these criteria in the
884
evaluation of their proposed models. Therefore, ML techniques are vital in the application of the
885
IoMT security. However, future studies should focus on how to use ML in a proper way to
886
concede the nature of the IoMT. 887 875
Moreover, the tools and environment of the current works are a combination of network
876
simulators and ML tools with more focus on the latter. Additionally, there is a lack of relevant
877
datasets, especially in the intrusion detection. Most of the studies focused on improving the
878
common ML algorithms evaluation metrics such as high accuracy and low FPR. However, since
879
the IoMT devices are characterized with shortage in power and small memory and processor,
880
there should be a balance between security and maintaining resources lifespan during the
881
adaption of these solutions. We have concluded that traditional ML techniques may fail if proper
882
consideration is not given to some metrics such as resource complexity, time complexity, and
883
energy usage. It was noticed that a vast majority of the studies ignored these criteria in the
884
evaluation of their proposed models. Therefore, ML techniques are vital in the application of the
885
IoMT security. However, future studies should focus on how to use ML in a proper way to
886
concede the nature of the IoMT. 888
References Journal of Ambient
908
Intelligence and Humanized Computing:1-17. 909
Alassaf N, and Gutub A. 2019. Simulating Light-Weight-Cryptography Implementation for IoT Healthcare
910
Data Security Applications. International Journal of E-Health and Medical Communications 10:1-
911
15. 10.4018/ijehmc.2019100101 909
Alassaf N, and Gutub A. 2019. Simulating Light-Weight-Cryptography Implementation for IoT Healthcare
910
Data Security Applications. International Journal of E-Health and Medical Communications 10:1-
911
15. 10.4018/ijehmc.2019100101 912
Alimadadi A, Aryal S, Manandhar I, Munroe PB, Joe B, and Cheng X. 2020. Artificial Intelligence and
913
Machine Learning to Fight COVID-19. American Physiological Society Rockville, MD. 912
Alimadadi A, Aryal S, Manandhar I, Munroe PB, Joe B, and Cheng X. 2020. Artificial Intelligence and
913
Machine Learning to Fight COVID-19. American Physiological Society Rockville, MD. 914
Alrashdi I, Alqazzaz A, Alharthi R, Aloufi E, Zohdy MA, and Ming H. 2019. FBAD: Fog-based attack
915
detection for IoT healthcare in smart cities. 2019 IEEE 10th Annual Ubiquitous Computing,
916
Electronics & Mobile Communication Conference (UEMCON): IEEE. p 0515-0522. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 942
Barros A, Rosário D, Resque P, and Cerqueira E. 2019. Heart of IoT: ECG as biometric sign for
943
authentication and identification. 2019 15th International Wireless Communications & Mobile
944
Computing Conference (IWCMC): IEEE. p 307-312. 942
Barros A, Rosário D, Resque P, and Cerqueira E. 2019. Heart of IoT: ECG as biometric sign for
943
authentication and identification. 2019 15th International Wireless Communications & Mobile
944
Computing Conference (IWCMC): IEEE. p 307-312. 945
Begli M, Derakhshan F, and Karimipour H. 2019. A layered intrusion detection system for critical
946
infrastructure using machine learning. 2019 IEEE 7th International Conference on Smart Energy
947
Grid Engineering (SEGE): IEEE. p 120-124. 945
Begli M, Derakhshan F, and Karimipour H. 2019. A layered intrusion detection system for critical
946
infrastructure using machine learning. 2019 IEEE 7th International Conference on Smart Energy
947
Grid Engineering (SEGE): IEEE. p 120-124. 948
Ben Amor L, Lahyani I, and Jmaiel M. 2020. AUDIT: AnomaloUs data Detection and Isolation approach
949
for mobile healThcare systems. Expert Systems 37:e12390. 950
Bostami B, Ahmed M, and Choudhury S. 2019. False Data Injection Attacks in Internet of Things. 951
Performability in Internet of Things: Springer, 47-58. 952
Bostani H, and Sheikhan M. 2017. Hybrid of anomaly-based and specification-based IDS for Internet of
953
Things using unsupervised OPF based on MapReduce approach. Computer Communications
954
98:52-71. 955
Burhan M, Rehman RA, Khan B, and Kim B-S. 2018. IoT elements, layered architectures and security
956
issues: A comprehensive survey. Sensors 18:2796. 957
Butun I, Kantarci B, and Erol-Kantarci M. 2015. Anomaly detection and privacy preservation in cloud-
958
centric Internet of Things. 2015 IEEE International Conference on Communication Workshop
959
(ICCW): IEEE. p 2610-2615. 2017. Data science: a comprehensive overview. ACM Computing Surveys (CSUR) 50:1-42. 960
Cao L. 2017. Data science: a comprehensive overview. ACM Computing Surveys (C 960
Cao L. 2017. Data science: a comprehensive overview. ACM Computing Surveys (CSUR) 50:1-42. 961
Chakraborty A, Alam M, Dey V, Chattopadhyay A, and Mukhopadhyay D. 2018. Adversarial attacks and
962
defences: A survey. arXiv preprint arXiv:181000069. 961
Chakraborty A, Alam M, Dey V, Chattopadhyay A, and Mukhopadhyay D. 2018. Adversarial attacks and
962
defences: A survey. arXiv preprint arXiv:181000069. 961
Chakraborty A, Alam M, Dey V, Chattopadhyay A, and Mukhopa
962
defences: A survey. arXiv preprint arXiv:181000069. 963
Cui L, Yang S, Chen F, Ming Z, Lu N, and Qin J. 2018. Manuscript to be reviewed 917
Alsubaei F, Abuhussein A, and Shiva S. 2019a. A Framework for Ranking IoMT Solutions Based on
918
Measuring Security and Privacy. In: Arai K, Bhatia R, and Kapoor S, eds. Proceedings of the Future
919
Technologies Conference. Cham: Springer International Publishing Ag, 205-224. 919
Technologies Conference. Cham: Springer International Publishing Ag, 205-224. 920
Alsubaei F, Abuhussein A, and Shiva S. 2019b. Ontology-Based Security Recommendation for the
921
Internet of Medical Things. IEEE Access 7:48948-48960. 10.1109/access.2019.2910087 920
Alsubaei F, Abuhussein A, and Shiva S. 2019b. Ontology-Based Security Recommendation for the
921
Internet of Medical Things. IEEE Access 7:48948-48960. 10.1109/access.2019.2910087 922
Aluvalu R. 2020. Intrusion Detection System for the IoT: A Comprehensive Review. Proceedings of the
923
11th International Conference on Soft Computing and Pattern Recognition (SoCPaR 2019):
924
Springer Nature. p 235. 925
Aman AHM, Hassan WH, Sameen S, Attarbashi ZS, Alizadeh M, and Latiff LA. 2020. IoMT amid COVID-19
926
pandemic: Application, architecture, technology, and security. Journal of Network and Computer
927
Applications:102886. 925
Aman AHM, Hassan WH, Sameen S, Attarbashi ZS, Alizadeh M, and Latiff LA. 2020. IoMT amid COVID-19
926
pandemic: Application, architecture, technology, and security. Journal of Network and Computer
927
Applications:102886. 928
Amanullah MA, Habeeb RAA, Nasaruddin FH, Gani A, Ahmed E, Nainar ASM, Akim NM, and Imran M. 929
2020. Deep learning and big data technologies for IoT security. Computer Communications
930
151:495-517. Williams L, and Burnap P. 2018. Pulse: an adaptive intrusion detection for the internet of t 31
Anthi E, Williams L, and Burnap P. 2018. Pulse: an adaptive intrusion detection for th 932
Ardabili SF, Mosavi A, Ghamisi P, Ferdinand F, Varkonyi-Koczy AR, Reuter U, Rabczuk T, and Atkinson
933
PM. 2020. Covid-19 outbreak prediction with machine learning. Available at SSRN 3580188. 934
Arshad J, Azad MA, Amad R, Salah K, Alazab M, and Iqbal R. 2020. A Review of Performance, Energy and
935
Privacy of Intrusion Detection Systems for IoT. Electronics 9:629. 936
Asfaw B, Bekele D, Eshete B, Villafiorita A, and Weldemariam K. 2010. Host-based anomaly detection for
937
pervasive medical systems. 2010 Fifth International Conference on Risks and Security of Internet
938
and Systems (CRiSIS): IEEE. p 1-8. 939
Bahşi H, Nõmm S, and La Torre FB. 2018. Dimensionality reduction for machine learning based iot botnet
940
detection. 2018 15th International Conference on Control, Automation, Robotics and Vision
941
(ICARCV): IEEE. p 1857-1862. Manuscript to be reviewed A survey on application of machine learning for
964
Internet of Things. International Journal of Machine Learning and Cybernetics 9:1399-1417. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 994
Farroha J. 2019. Security Analysis and Recommendations for AI/ML Enabled Automated Cyber Medical
995
Systems. In: Ahmad F, ed. Big Data: Learning, Analytics, and Applications. Bellingham: Spie-Int
996
Soc Optical Engineering. 997
Fernandez Maimo L, Huertas Celdran A, Perales Gomez AL, Garcia Clemente FJ, Weimer J, and Lee I. 998
2019. Intelligent and dynamic ransomware spread detection and mitigation in integrated clinica
999
environments. Sensors 19:1114. 1000
Firouzi F, Rahmani AM, Mankodiya K, Badaroglu M, Merrett GV, Wong P, and Farahani B. 2018. Internet
1001
of-Things and big data for smarter healthcare: From device to architecture, applications and
1002
analytics. Elsevier. 1003
Gao S, and Thamilarasu G. 2017. Machine-learning classifiers for security in connected medical devices. 1004
2017 26th International Conference on Computer Communication and Networks (ICCCN): IEEE. p
1005
1-5. 1006
Gheisari M, Wang G, and Bhuiyan MZA. 2017. A survey on deep learning in big data. 2017 IEEE
1007
international conference on computational science and engineering (CSE) and IEEE internationa
1008
conference on embedded and ubiquitous computing (EUC): IEEE. p 173-180. 1009
Grammatikis PIR, Sarigiannidis PG, and Moscholios ID. 2019. Securing the Internet of Things: Challenges,
1010
threats and solutions. Internet of Things 5:41-70. 1011
Gupta R, Tanwar S, Tyagi S, and Kumar N. 2020a. Machine learning models for secure data analytics: A
1012
taxonomy and threat model. Computer Communications 153:406-440. 994
Farroha J. 2019. Security Analysis and Recommendations for AI/ML Enabled Automated Cyber Medical
995
Systems. In: Ahmad F, ed. Big Data: Learning, Analytics, and Applications. Bellingham: Spie-Int
996
Soc Optical Engineering. 997
Fernandez Maimo L, Huertas Celdran A, Perales Gomez AL, Garcia Clemente FJ, Weimer J, and Lee I. 998
2019. Intelligent and dynamic ransomware spread detection and mitigation in integrated clinical
999
environments. Sensors 19:1114. 1000
Firouzi F, Rahmani AM, Mankodiya K, Badaroglu M, Merrett GV, Wong P, and Farahani B. 2018. Internet-
1001
of-Things and big data for smarter healthcare: From device to architecture, applications and
1002
analytics. Elsevier. 1003
Gao S, and Thamilarasu G. 2017. Machine-learning classifiers for security in connected medical devices. 1004
2017 26th International Conference on Computer Communication and Networks (ICCCN): IEEE. p
1005
1-5. 1006
Gheisari M, Wang G, and Bhuiyan MZA. 2017. A survey on deep learning in big data. 2017 IEEE
1007
international conference on computational science and engineering (CSE) and IEEE international
1008
conference on embedded and ubiquitous computing (EUC): IEEE. p 173-180. Manuscript to be reviewed 965
da Costa KA, Papa JP, Lisboa CO, Munoz R, and de Albuquerque VHC. 2019. Internet of Things: A survey
966
on machine learning-based intrusion detection approaches. Computer Networks 151:147-157. 965
da Costa KA, Papa JP, Lisboa CO, Munoz R, and de Albuquerque VHC. 2019. Internet of Things: A survey
966
on machine learning-based intrusion detection approaches. Computer Networks 151:147-157. 967
Dang LM, Piran M, Han D, Min K, and Moon H. 2019. A survey on internet of things and cloud computing
968
for healthcare. Electronics 8:768. 969
Davis J. 2020a. Ransomware Attacks Cost Healthcare Sector At Least $160M Since 2016. Available at
970
https://healthitsecurity.com/news/ransomware-attacks-cost-healthcare-sector-at-least-160m-
971
since-2016 (accessed 11/1/2020. 972
Davis J. 2020b. US Ransomware Attacks Doubled in Q3; Healthcare Sector Most Targeted. Available at
973
https://healthitsecurity.com/news/us-ransomware-attacks-doubled-in-q3-healthcare-sector-
974
most-targeted (accessed 2/1/2021. 975
Dimitrov DV. 2016. Medical internet of things and big data in healthcare. Healthcare informatics
976
research 22:156-163. 975
Dimitrov DV. 2016. Medical internet of things and big data in healthcare. Healthcare informatics
976
research 22:156-163. 977
Din IU, Almogren A, Guizani M, and Zuair M. 2019. A Decade of Internet of Things: Analysis in the Light
978
of Healthcare Applications. IEEE Access 7:89967-89979. 10.1109/access.2019.2927082 979
Diro AA, and Chilamkurti N. 2018. Distributed attack detection scheme using deep learning approach for
980
Internet of Things. Future Generation Computer Systems 82:761-768. 981
Durga S, Nag R, and Daniel E. 2019. Survey on machine learning and deep learning algorithms used in
982
internet of things (IoT) healthcare. 2019 3rd International Conference on Computing
983
Methodologies and Communication (ICCMC): IEEE. p 1018-1022. 984
Elrawy MF, Awad AI, and Hamed HF. 2018. Intrusion detection systems for IoT-based smart
985
environments: a survey. Journal of Cloud Computing 7:21. 986
Escamilla-Ambrosio P, Rodríguez-Mota A, Aguirre-Anaya E, Acosta-Bermejo R, and Salinas-Rosales M. 987
2018. Distributing computing in the internet of things: cloud, fog and edge computing overview. 988
NEO 2016: Springer, 87-115. 989
Eskandari M, Janjua ZH, Vecchio M, and Antonelli F. 2020. Passban IDS: An intelligent anomaly based
990
intrusion detection system for IoT edge devices. IEEE Internet of Things Journal. 991
Farahani B, Firouzi F, Chang V, Badaroglu M, Constant N, and Mankodiya K. 2018. Towards fog-driven IoT
992
eHealth: Promises and challenges of IoT in medicine and healthcare. Future Generation
993
Computer Systems 78:659-676. 992
eHealth: Promises and challenges of IoT in medicine and healthcare. Future Generation
993
Computer Systems 78:659-676. Manuscript to be reviewed 1009
Grammatikis PIR, Sarigiannidis PG, and Moscholios ID. 2019. Securing the Internet of Things: Challenges,
1010
threats and solutions. Internet of Things 5:41-70. 1011
Gupta R, Tanwar S, Tyagi S, and Kumar N. 2020a. Machine learning models for secure data analytics: A
1012
taxonomy and threat model. Computer Communications 153:406-440. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 1031
He D, Qiao Q, Gao Y, Zheng J, Chan S, Li J, and Guizani N. 2019. Intrusion Detection Based on Stacked
1032
Autoencoder for Connected Healthcare Systems. IEEE Network 33:64-69. 1033
He D, Ye R, Chan S, Guizani M, and Xu Y. 2018. Privacy in the Internet of Things for smart healthcare. 1034
IEEE Communications Magazine 56:38-44. 1033
He D, Ye R, Chan S, Guizani M, and Xu Y. 2018. Privacy in the Internet of Things for smart healthcare. 1034
IEEE Communications Magazine 56:38-44. 1035
Hei X, Du X, Wu J, and Hu F. 2010. Defending resource depletion attacks on implantable medical devices. 1036
2010 IEEE global telecommunications conference GLOBECOM 2010: IEEE. p 1-5. 1035
Hei X, Du X, Wu J, and Hu F. 2010. Defending resource depletion attacks on implantable medical devices. 1036
2010 IEEE global telecommunications conference GLOBECOM 2010: IEEE. p 1-5. 1037
Hussain F, Hussain R, Hassan SA, and Hossain E. 2020. Machine learning in IoT security: current solutions
1038
and future challenges. IEEE Communications Surveys & Tutorials. 1037
Hussain F, Hussain R, Hassan SA, and Hossain E. 2020. Machine learning in IoT security: current solutions
1038
and future challenges. IEEE Communications Surveys & Tutorials. 1039
Ibitoye O, Shafiq O, and Matrawy A. 2019. Analyzing adversarial attacks against deep learning for
1040
intrusion detection in IoT networks. 2019 IEEE Global Communications Conference
1041
(GLOBECOM): IEEE. p 1-6. 1039
Ibitoye O, Shafiq O, and Matrawy A. 2019. Analyzing adversarial attacks against deep learning for
1040
intrusion detection in IoT networks. 2019 IEEE Global Communications Conference
1041
(GLOBECOM): IEEE. p 1-6. 1042
Itten A, and Vadakkumcheril GT. 2016. ENHANCED INTRUSION DETECTION SYSTEM IN MEDICAL CYBER
1043
PHYSICAL SYSTEMS USING MULTIVARIATE CORRELATION ANALYSIS. 1042
Itten A, and Vadakkumcheril GT. 2016. ENHANCED INTRUSION DETECTION SYSTEM IN MEDICAL CYBER
1043
PHYSICAL SYSTEMS USING MULTIVARIATE CORRELATION ANALYSIS. 1044
Jaigirdar FT, Rudolph C, and Bain C. 2019. Can I Trust the Data I See? A Physician's Concern on Medical
1045
Data in IoT Health Architectures. Proceedings of the Australasian Computer Science Week
1046
Multiconference. p 1-10. 1044
Jaigirdar FT, Rudolph C, and Bain C. 2019. Can I Trust the Data I See? A Physician's Concern on Medical
1045
Data in IoT Health Architectures. Proceedings of the Australasian Computer Science Week
1046
Multiconference. p 1-10. 1047
Jan SU, Ahmed S, Shakhov V, and Koo I. 2019. Manuscript to be reviewed 1013
Gupta S, Venugopal V, Mahajan V, Gaur S, Barnwal M, and Mahajan H. 2020b. HIPAA, GDPR and Best
1014
Practice Guidelines for preserving data security and privacy-What Radiologists should know. 1015
European Congress of Radiology-ECR 2020. 1013
Gupta S, Venugopal V, Mahajan V, Gaur S, Barnwal M, and Mahajan H. 2020b. HIPAA, GDPR and Best
1014
Practice Guidelines for preserving data security and privacy-What Radiologists should know. 1015
European Congress of Radiology-ECR 2020. 1016
Hameed SS, Hassan R, and Muhammad FF. 2017. Selection and classification of gene expression in
1017
autism disorder: Use of a combination of statistical filters and a GBPSO-SVM algorithm. PloS one
1018
12. 1019
Haque SA, Rahman M, and Aziz SM. 2015. Sensor anomaly detection in wireless sensor networks for
1020
healthcare. Sensors 15:8764-8786. 1019
Haque SA, Rahman M, and Aziz SM. 2015. Sensor anomaly detection in wireless sensor networks for
1020
healthcare. Sensors 15:8764-8786. 1021
Hasan M, Islam MM, Zarif MII, and Hashem M. 2019. Attack and anomaly detection in IoT sensors in IoT
1022
sites using machine learning approaches. Internet of Things 7:100059. 1021
Hasan M, Islam MM, Zarif MII, and Hashem M. 2019. Attack and anomaly detection in IoT sensors in IoT
1022
sites using machine learning approaches. Internet of Things 7:100059. 1023
Hassija V, Chamola V, Saxena V, Jain D, Goyal P, and Sikdar B. 2019. A survey on IoT security: application
1024
areas, security threats, and solution architectures. IEEE Access 7:82721-82743. 1025
Hatzivasilis G, Soultatos O, Ioannidis S, Verikoukis C, Demetriou G, Tsatsoulis CI, and Ieee. 2019. Review
1026
of Security and Privacy for the Internet of Medical Things (IoMT) Resolving the protection
1027
concerns for the novel circular economy bioinformatics. 2019 15th International Conference on
1028
Distributed Computing in Sensor Systems. New York: Ieee, 457-464. 1029
Hau Z, and Lupu EC. 2019. Exploiting Correlations to Detect False Data Injections in Low-Density Wireless
1030
Sensor Networks. Proceedings of the 5th on Cyber-Physical System Security Workshop. p 1-12. 1031
He D, Qiao Q, Gao Y, Zheng J, Chan S, Li J, and Guizani N. 2019. Intrusion Detection Based on Stacked
1032
Autoencoder for Connected Healthcare Systems IEEE Network 33:64 69 1029
Hau Z, and Lupu EC. 2019. Exploiting Correlations to Detect False Data Injections in Low-Density Wireless
1030
Sensor Networks. Proceedings of the 5th on Cyber-Physical System Security Workshop. p 1-12. Manuscript to be reviewed 1060
Liang W, Huang W, Long J, Zhang K, Li K-C, and Zhang D. 2020. Deep reinforcement learning for resource
1061
protection and real-time detection in IoT environment. IEEE Internet of Things Journal. 1060
Liang W, Huang W, Long J, Zhang K, Li K-C, and Zhang D. 2020. Deep reinforcement learning for resource
1061
protection and real-time detection in IoT environment. IEEE Internet of Things Journal. 1062
Liu CH, Lin Q, and Wen S. 2018. Blockchain-enabled data collection and sharing for industrial IoT with
1063
deep reinforcement learning. IEEE Transactions on Industrial Informatics 15:3516-3526. 1062
Liu CH, Lin Q, and Wen S. 2018. Blockchain-enabled data collection and sharing for industrial IoT with
1063
deep reinforcement learning. IEEE Transactions on Industrial Informatics 15:3516-3526. 1064
Lopez-Martin M, Carro B, Sanchez-Esguevillas A, and Lloret J. 2017. Conditional variational autoencoder
1065
for prediction and feature recovery applied to intrusion detection in iot. Sensors 17:1967. 1064
Lopez-Martin M, Carro B, Sanchez-Esguevillas A, and Lloret J. 2017. Conditional variational autoencoder
1065
for prediction and feature recovery applied to intrusion detection in iot. Sensors 17:1967. 1066
Ma Y, Wang Y, Yang J, Miao Y, and Li W. 2017. Big Health Application System based on Health Internet of
1067
Things and Big Data. IEEE Access 5:7885-7897. 10.1109/ACCESS.2016.2638449 1066
Ma Y, Wang Y, Yang J, Miao Y, and Li W. 2017. Big Health Application System based on Health Internet of
1067
Things and Big Data. IEEE Access 5:7885-7897. 10.1109/ACCESS.2016.2638449 1068
Maleh Y, Ezzati A, Qasmaoui Y, and Mbida M. 2015. A global hybrid intrusion detection system for
1069
wireless sensor networks. Procedia Computer Science 52:1047-1052. 1070
Manimurugan S, Almutairi S, Aborokbah MM, Chilamkurti N, Ganesan S, and Patan R. 2020. Effective
1071
Attack Detection in Internet of Medical Things Smart Environment using a Deep Belief Neural
1072
Network. IEEE Access. 1073
Marjani M, Nasaruddin F, Gani A, Karim A, Hashem IAT, Siddiqa A, and Yaqoob I. 2017. Big IoT data
1074
analytics: architecture, opportunities, and open research challenges. IEEE Access 5:5247-5261. 1075
Mawgoud AA, Karadawy AI, and Tawfik BS. 2019. A Secure Authentication Technique in Internet of
1076
Medical Things through Machine Learning. arXiv preprint arXiv:191212143. 1077
McMahon E, Williams R, El M, Samtani S, Patton M, and Chen H. 2017. Assessing medical device
1078
vulnerabilities on the Internet of Things. 2017 IEEE International Conference on Intelligence and
1079
Security Informatics (ISI): IEEE. p 176-178. Manuscript to be reviewed Toward a lightweight intrusion detection system for the
1048
internet of things. IEEE Access 7:42450-42471. 1047
Jan SU, Ahmed S, Shakhov V, and Koo I. 2019. Toward a lightweight intrusion detection system for the
1048
internet of things. IEEE Access 7:42450-42471. 1049
Khan FA, Haldar NAH, Ali A, Iftikhar M, Zia TA, and Zomaya AY. 2017. A continuous change detection
1050
mechanism to identify anomalies in ECG signals for WBAN-based healthcare environments. IEEE
1051
Access 5:13531-13544. 1049
Khan FA, Haldar NAH, Ali A, Iftikhar M, Zia TA, and Zomaya AY. 2017. A continuous change detection
1050
mechanism to identify anomalies in ECG signals for WBAN-based healthcare environments. IEEE
1051
Access 5:13531-13544. 1052
Kintzlinger M, Cohen A, Nissim N, Rav-Acha M, Khalameizer V, Elovici Y, Shahar Y, and Katz A. 2020. 1053
CardiWall: A Trusted Firewall for the Detection of Malicious Clinical Programming of Cardiac
1054
Implantable Electronic Devices IEEE Access 8:48123 48140 1055
Kubat M. 2017. An introduction to machine learning: Springer. 1055
Kubat M. 2017. An introduction to machine learning: Springer. 1056
Landau O, Cohen A, Gordon S, and Nissim N. 2020. Mind your privacy: Privacy leakage through BCI
1057
applications using machine learning methods. Knowledge-Based Systems:105932. 1058
Li Q, Li W, Wang J, and Cheng M. 2019. A SQL Injection Detection Method Based on Adaptive Deep
1059
Forest. IEEE Access 7:145385-145394. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 1080
Meidan Y, Bohadana M, Mathov Y, Mirsky Y, Shabtai A, Breitenbacher D, and Elovici Y. 2018. N-BaIoT–
1081
network-based detection of IoT botnet attacks using deep autoencoders. IEEE Pervas Comput 17
1082
(3): 12–22. 1083
Midi D, Rullo A, Mudgerikar A, and Bertino E. 2017. Kalis—A system for knowledge-driven adaptable
1084
intrusion detection for the Internet of Things. 2017 IEEE 37th International Conference on
1085
Distributed Computing Systems (ICDCS): IEEE. p 656-666. 1086
Miyato T, Maeda S-i, Koyama M, and Ishii S. 2018. Virtual adversarial training: a regularization method
1087
for supervised and semi-supervised learning. IEEE transactions on pattern analysis and machine
1088
intelligence 41:1979-1993. 1089
Mohamed MB, Meddeb-Makhlouf A, and Fakhfakh A. 2019. Intrusion cancellation for anomaly detection
1090
in healthcare applications. 2019 15th International Wireless Communications & Mobile
1091
Computing Conference (IWCMC): IEEE. p 313-318. 1092
Mohsen NR, Ying B, and Nayak A. 2019. Authentication Protocol for Real-Time Wearable Medical Sensor
1093
Networks Using Biometrics and Continuous Monitoring. 2019 International Conference on
1094
Internet of Things (iThings) and IEEE Green Computing and Communications (GreenCom) and
1095
IEEE Cyber, Physical and Social Computing (CPSCom) and IEEE Smart Data (SmartData): IEEE. p
1096
1199-1206. 1097
Mohsin A, Zaidan A, Zaidan B, Albahri O, Albahri A, Alsalem M, and Mohammed K. 2019. Based
1098
blockchain-PSO-AES techniques in finger vein biometrics: A novel verification secure framework
1099
for patient authentication. Computer Standards & Interfaces 66:103343. 1100
Mosenia A, and Jha NK. 2016. A comprehensive study of security of internet-of-things. IEEE Transactions
1101
on Emerging Topics in Computing 5:586-602. 1102
Mozaffari-Kermani M, Sur-Kolay S, Raghunathan A, and Jha NK. 2014. Systematic poisoning attacks on
1103
and defenses for machine learning in healthcare. IEEE journal of biomedical and health
1104
informatics 19:1893-1905. 1105
Musale P, Baek D, Werellagama N, Woo SS, and Choi BJ. 2019. You walk, we authenticate: Lightweight
1106
seamless authentication based on gait in wearable IoT systems. IEEE Access 7:37883-37895. 1105
Musale P, Baek D, Werellagama N, Woo SS, and Choi BJ. 2019. You walk, we authenticate: Lightweight
1106
seamless authentication based on gait in wearable IoT systems. IEEE Access 7:37883-37895. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 1107
Nagdeo SK, and Mahapatro J. 2019. Wireless Body Area Network Sensor Faults and Anomalous Data
1108
Detection and Classification using Machine Learning. 2019 IEEE Bombay Section Signature
1109
Conference (IBSSC): IEEE. p 1-6. 1107
Nagdeo SK, and Mahapatro J. 2019. Wireless Body Area Network Sensor Faults and Anomalous Data
1108
Detection and Classification using Machine Learning. 2019 IEEE Bombay Section Signature
1109
Conference (IBSSC): IEEE. p 1-6. 1110
Nanayakkara N, Halgamuge M, and Syed A. 2019. Security and Privacy of Internet of Medical Things
1111
(IoMT) Based Healthcare Applications: A Review. 1110
Nanayakkara N, Halgamuge M, and Syed A. 2019. Security and Privacy of Internet of Medical Things
1111
(IoMT) Based Healthcare Applications: A Review. 1112
Newaz A, Sikder AK, Rahman MA, and Uluagac AS. 2020. A Survey on Security and Privacy Issues in
1113
Modern Healthcare Systems: Attacks and Defenses. arXiv preprint arXiv:200507359. 1112
Newaz A, Sikder AK, Rahman MA, and Uluagac AS. 2020. A Survey on Security and Privacy Issues in
1113
Modern Healthcare Systems: Attacks and Defenses. arXiv preprint arXiv:200507359. 1114
Newaz AI, Sikder AK, Rahman MA, and Uluagac AS. 2019. Healthguard: A machine learning-based
1115
security framework for smart healthcare systems. 2019 Sixth International Conference on Social
1116
Networks Analysis, Management and Security (SNAMS): IEEE. p 389-396. 1114
Newaz AI, Sikder AK, Rahman MA, and Uluagac AS. 2019. Healthguard: A machine learning-based
1115
security framework for smart healthcare systems. 2019 Sixth International Conference on Social
1116
Networks Analysis, Management and Security (SNAMS): IEEE. p 389-396. 1117
Nguyen DC, Pathirana PN, Ding M, and Seneviratne A. 2019. Blockchain for Secure EHRs Sharing of
1118
Mobile Cloud Based E-Health Systems. IEEE Access 7:66792-66806. 1119
10.1109/access.2019.2917555 1117
Nguyen DC, Pathirana PN, Ding M, and Seneviratne A. 2019. Blockchain for Secure EHRs Sharing of
1118
Mobile Cloud Based E-Health Systems. IEEE Access 7:66792-66806. / 1120
Noor MM, and Hassan WH. 2019. Current research on Internet of Things (IoT) security: A survey. 1121
Computer Networks 148:283-294. 1120
Noor MM, and Hassan WH. 2019. Current research on Internet of Things (IoT) security: A survey. 1121
Computer Networks 148:283-294. 1122
Odesile A, and Thamilarasu G. 2017. Distributed intrusion detection using mobile agents in wireless body
1123
area networks. 2017 Seventh International Conference on Emerging Security Technologies (EST):
1124
IEEE. p 144-149. 1125
Pandey P, and Litoriya R. 2020. Manuscript to be reviewed 1170
Sadoughi F, Behmanesh A, and Sayfouri N. 2020. Internet of Things in Medicine: A Systematic Mapping
1171
Study. Journal of Biomedical Informatics:103383. 1172
Saeed A, Ahmadinia A, Javed A, and Larijani H. 2016. Random neural network based intelligent intrusion
1173
detection for wireless sensor networks. Procedia Computer Science 80:2372-2376. 1172
Saeed A, Ahmadinia A, Javed A, and Larijani H. 2016. Random neural network based intelligent intrusion
1173
detection for wireless sensor networks. Procedia Computer Science 80:2372-2376. 1174
Saheb T, and Izadi L. 2019. Paradigm of IoT big data analytics in healthcare industry: a review of scientific
1175
literature and mapping of research trends. Telematics and Informatics. 1174
Saheb T, and Izadi L. 2019. Paradigm of IoT big data analytics in healthcare industry: a review of scientific
1175
literature and mapping of research trends. Telematics and Informatics. 1176
Salem O, Guerassimov A, Mehaoua A, Marcus A, and Furht B. 2014. Anomaly detection in medical
1177
wireless sensor networks using SVM and linear regression models. International Journal of E-
1178
Health and Medical Communications (IJEHMC) 5:20-45. 1176
Salem O, Guerassimov A, Mehaoua A, Marcus A, and Furht B. 2014. Anomaly detection in medical
1177
wireless sensor networks using SVM and linear regression models. International Journal of E-
1178
Health and Medical Communications (IJEHMC) 5:20-45. 1179
Santagati GE, Dave N, and Melodia T. 2020. Design and Performance Evaluation of an Implantable
1180
Ultrasonic Networking Platform for the Internet of Medical Things. IEEE/ACM Transactions on
1181
Networking 28:29-42. 10.1109/TNET.2019.2949805 1179
Santagati GE, Dave N, and Melodia T. 2020. Design and Performance Evaluation of an Implantable
1180
Ultrasonic Networking Platform for the Internet of Medical Things. IEEE/ACM Transactions on
1181
Networking 28:29-42. 10.1109/TNET.2019.2949805 1182
Schneble W, and Thamilarasu G. 2019a. Attack Detection Using Federated Learning in Medical Cyber-
1183
Physical Systems. 1182
Schneble W, and Thamilarasu G. 2019a. Attack Detection Using Federated Learning in Medical Cyber-
1183
Physical Systems. 1184
Schneble W, and Thamilarasu G. 2019b. Attack Detection Using Federated Learning in Medical Cyber-
1185
Physical Systems. IEEE 1186
Schneble W, and Thamilarasu G. 2019c. Optimal Feature Selection for Intrusion Detection in Medical
1187
Cyber-Physical Systems. 2019 11th International Conference on Advanced Computing (ICoAC):
1188
IEEE. p 238-243. 1189
Sehatbakhsh N, Alam M, Nazari A, Zajic A, and Prvulovic M. 2018. Syndrome: Spectral analysis for
1190
anomaly detection on medical iot and embedded devices. Manuscript to be reviewed Elderly care through unusual behavior detection: A disaster management
1126
approach using IoT and intelligence. IBM Journal of Research and Development 64:15:11-15:11. 1127
10.1147/JRD.2019.2947018 1128
Pandey R, Gautam V, Bhagat K, and Sethi T. 2020. A Machine Learning Application for Raising WASH
1129
Awareness in the Times of Covid-19 Pandemic. arXiv preprint arXiv:200307074. Pannu A. 2015. Artificial intelligence and its application in different areas. Artificial Intellig 1130
Pannu A. 2015. Artificial intelligence and its application in different areas. Artificial Intelligence 4:79-84. g
pp
f
g
1131
Parra GDLT, Rad P, Choo K-KR, and Beebe N. 2020. Detecting Internet of Things attacks using distributed
1132
deep learning. Journal of Network and Computer Applications:102662. 1133
Patel N. 2020. Social Engineering As an Evolutionary Threat to Information Security in Healthcare
1134
Organizations. Jurnal Administrasi Kesehatan Indonesia 8. 1135
Pirbhulal S, Pombo N, Felizardo V, Garcia N, Sodhro AH, and Mukhopadhyay SC. 2019. Towards Machine
1136
Learning Enabled Security Framework for IoT-based Healthcare. 2019 13th International
1137
Conference on Sensing Technology (ICST): IEEE. p 1-6. 1138
Pramanik MI, Lau RY, Demirkan H, and Azad MAK. 2017. Smart health: Big data enabled health paradigm
1139
within smart cities. Expert Systems with Applications 87:370-383. 1140
Punithavathi P, Geetha S, Karuppiah M, Islam SH, Hassan MM, and Choo K-KR. 2019. A lightweight
1141
machine learning-based authentication framework for smart IoT devices. Information Sciences
1142
484:255-268. 1143
Qu G, and Yuan L. 2014. Design THINGS for the Internet of Things—An EDA perspective. 2014 IEEE/ACM
1144
international conference on Computer-Aided Design (ICCAD): IEEE. p 411-416. 1145
Rahman MA, and Mohsenian-Rad H. 2012. False data injection attacks with incomplete information
1146
against smart power grids. 2012 IEEE Global Communications Conference (GLOBECOM): IEEE. p
1147
3153-3158. 1148
Rahmani AM, Gia TN, Negash B, Anzanpour A, Azimi I, Jiang M, and Liljeberg P. 2018. Exploiting smart e-
1149
Health gateways at the edge of healthcare Internet-of-Things: A fog computing approach. Future
1150
Generation Computer Systems 78:641-658. 1151
Rathore H, Al-Ali A, Mohamed A, Du X, and Guizani M. 2018a. DTW based authentication for wireless
1152
medical device security. 2018 14th International Wireless Communications & Mobile Computing
1153
Conference (IWCMC): IEEE. p 476-481. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 1154
Rathore H, Al-Ali AK, Mohamed A, Du X, and Guizani M. 2019. A Novel Deep Learning Strategy for
1155
Classifying Different Attack Patterns for Deep Brain Implants. IEEE Access 7:24154-24164. 1156
Rathore H, Fu C, Mohamed A, Al-Ali A, Du X, Guizani M, and Yu Z. 2018b. Multi-layer security scheme for
1157
implantable medical devices. Neural Computing and Applications:1-14. 1156
Rathore H, Fu C, Mohamed A, Al-Ali A, Du X, Guizani M, and Yu Z. 2018b. Multi-layer security scheme fo
1157
implantable medical devices. Neural Computing and Applications:1-14. 1158
Rathore H, Wenzel L, Al-Ali AK, Mohamed A, Du X, and Guizani M. 2018c. Multi-layer perceptron model
1159
on chip for secure diabetic treatment. IEEE Access 6:44718-44730. 1158
Rathore H, Wenzel L, Al-Ali AK, Mohamed A, Du X, and Guizani M. 2018c. Multi-layer perceptron model
1159
on chip for secure diabetic treatment. IEEE Access 6:44718-44730. 1160
Rathore S, and Park JH. 2018. Semi-supervised learning based distributed attack detection framework
1161
for IoT. Applied Soft Computing 72:79-89. 1160
Rathore S, and Park JH. 2018. Semi-supervised learning based distributed attack detection framework
1161
for IoT. Applied Soft Computing 72:79-89. 1162
RM SP, Maddikunta PKR, Parimala M, Koppu S, Reddy T, Chowdhary CL, and Alazab M. 2020. An
1163
effective feature engineering for DNN using hybrid PCA-GWO for intrusion detection in IoMT
1164
architecture. Computer Communications. 1162
RM SP, Maddikunta PKR, Parimala M, Koppu S, Reddy T, Chowdhary CL, and Alazab M. 2020. An
1163
effective feature engineering for DNN using hybrid PCA-GWO for intrusion detection in IoMT
1164
architecture. Computer Communications. 1165
Roopak M, Tian GY, and Chambers J. 2020. Multi-objective-based feature selection for DDoS attack
1166
detection in IoT networks. IET Networks 9:120-127. 1165
Roopak M, Tian GY, and Chambers J. 2020. Multi-objective-based feature selection for DDoS attack
1166
detection in IoT networks. IET Networks 9:120-127. 1167
Russom P. 2011. Big data analytics. TDWI best practices report, fourth quarter 19:1-34. 1167
Russom P. 2011. Big data analytics. TDWI best practices report, fourth quarter 19:1-34. 1168
S. O'Dea S. 2020. Data volume of IoT connected devices worldwide 2018 and 2025. Available at
1169
https://www.statista.com/statistics/1017863/worldwide-iot-connected-devices-data-size/. 1168
S. O'Dea S. 2020. Data volume of IoT connected devices worldwide 2018 and 2025. Available at
1169
https://www.statista.com/statistics/1017863/worldwide-iot-connected-devices-data-size/. 1170
Sadoughi F, Behmanesh A, and Sayfouri N. 2020. Internet of Things in Medicine: A Systematic Mapping
1171
Study. Journal of Biomedical Informatics:103383. Manuscript to be reviewed 2018 IEEE international symposium
1191
on hardware oriented security and trust (HOST): IEEE. p 1-8. 1192
Sethi P, and Sarangi SR. 2017. Internet of things: architectures, protocols, and applications. Journal of
1193
Electrical and Computer Engineering 2017. 1194
Shafiq M, Tian Z, Bashir AK, Du X, and Guizani M. 2020. IoT malicious traffic identification using wrapper-
1195
based feature selection mechanisms. Computers & Security:101863. 1196
Shakeel PM, Baskar S, Dhulipala VS, Mishra S, and Jaber MM. 2018. Maintaining security and privacy in
1197
health care system using learning based deep-Q-networks. Journal of medical systems 42:186. 1198
Shang J, and Wu J. 2019. A Usable Authentication System Using Wrist-worn Photoplethysmography
1199
Sensors on Smartwatches. 2019 IEEE Conference on Communications and Network Security
1200
(CNS): IEEE. p 1-9. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 1201
Shen J, Chang S, Shen J, Liu Q, and Sun X. 2018. A lightweight multi-layer authentication protocol for
1202
wireless body area networks. Future Generation Computer Systems 78:956-963. 1201
Shen J, Chang S, Shen J, Liu Q, and Sun X. 2018. A lightweight multi-layer authentication protocol for
1202
wireless body area networks. Future Generation Computer Systems 78:956-963. 1201
Shen J, Chang S, Shen J, Liu Q, and Sun X. 2018. A lightweight multi-layer authentication protocol for
1202
wireless body area networks. Future Generation Computer Systems 78:956-963. 1204
Sollins KR. 2018. IoT Big Data Security and Privacy vs. Innovation. IEEE Internet Things J. 1204
Sollins KR. 2018. IoT Big Data Security and Privacy vs. Innovation. IEEE Internet Things J. 1205
Spiekermann S. 2015. Ethical IT innovation: A value-based system design appr 1206
Stiawan D, Idris M, Malik RF, Nurmaini S, Alsharif N, and Budiarto R. 2019. Investigating Brute Force
1207
Attack Patterns in IoT Network. Journal of Electrical and Computer Engineering 2019. 1206
Stiawan D, Idris M, Malik RF, Nurmaini S, Alsharif N, and Budiarto R. 2019. Investigating Brute Force
1207
Attack Patterns in IoT Network. Journal of Electrical and Computer Engineering 2019. 1208
Sun Y, Lo FP-W, and Lo B. 2019. Security and Privacy for the Internet of Medical Things Enabled
1209
Healthcare Systems: A Survey. IEEE Access 7:183339-183355. 1208
Sun Y, Lo FP-W, and Lo B. 2019. Security and Privacy for the Internet of Medical Things Enabled
1209
Healthcare Systems: A Survey. IEEE Access 7:183339-183355. 1210
Tahsien SM, Karimipour H, and Spachos P. 2020. Machine learning based solutions for security of
1211
Internet of Things (IoT): A survey. Journal of Network and Computer Applications:102630. 1210
Tahsien SM, Karimipour H, and Spachos P. 2020. Machine learning based solutions for security of
1211
Internet of Things (IoT): A survey. Journal of Network and Computer Applications:102630. 1212
Thamilarasu G. 2016. iDetect: an intelligent intrusion detection system for wireless body area networks. 1213
International Journal of Security and Networks 11:82-93. 1212
Thamilarasu G. 2016. iDetect: an intelligent intrusion detection system for wireless body area networks. 1213
International Journal of Security and Networks 11:82-93. 1214
Tovino SA. 2016. The HIPAA Privacy Rule and the EU GDPR: illustrative comparisons. Seton Hall L Rev
1215
47:973. 1214
Tovino SA. 2016. The HIPAA Privacy Rule and the EU GDPR: illustrative comparisons. Seton Hall L Rev
1215
47:973. 1216
Tseng TW, Wu CT, and Lai F. 2019. Manuscript to be reviewed Threat Analysis for Wearable Health Devices and Environment
1217
Monitoring Internet of Things Integration System. IEEE Access 7:144983-144994. 1218
10.1109/ACCESS.2019.2946081 1216
Tseng TW, Wu CT, and Lai F. 2019. Threat Analysis for Wearable Health Devices and Environment
1217
Monitoring Internet of Things Integration System. IEEE Access 7:144983-144994. 1218
10.1109/ACCESS.2019.2946081 1219
Verner A, and Butvinik D. 2017. A machine learning approach to detecting sensor data modification
1220
intrusions in WBANs. 2017 16th IEEE International Conference on Machine Learning and
1221
Applications (ICMLA): IEEE. p 161-169. 1222
Vhaduri S, and Poellabauer C. 2019. Multi-modal biometric-based implicit authentication of wearable
1223
device users. IEEE Transactions on Information Forensics and Security 14:3116-3125. 1224
Vinitha S, Sweetlin S, Vinusha H, and Sajini S. 2018. Disease prediction using machine learning over big
1225
data. Computer Science & Engineering: An International Jouranl (CSEIJ) 8. 1226
Wang Y, Meng W, Li W, Li J, Liu W-X, and Xiang Y. 2018. A fog-based privacy-preserving approach for
1227
distributed signature-based intrusion detection. Journal of Parallel and Distributed Computing
1228
122:26-35. 1229
Wazid M, Das AK, Rodrigues J, Shetty S, and Park Y. 2019a. IoMT Malware Detection Approaches:
1230
Analysis and Research Challenges. IEEE Access 7:182459-182476. 10.1109/access.2019.2960412 1229
Wazid M, Das AK, Rodrigues J, Shetty S, and Park Y. 2019a. IoMT Malware Detection Approaches:
1230
Analysis and Research Challenges. IEEE Access 7:182459-182476. 10.1109/access.2019.2960412
1231
Wazid M, Das AK, Rodrigues JJ, Shetty S, and Park Y. 2019b. IoMT malware detection approaches: 1230
Analysis and Research Challenges. IEEE Access 7:182459-182476. 10.1109/access.2019.2960412
1231
Wazid M, Das AK, Rodrigues JJ, Shetty S, and Park Y. 2019b. IoMT malware detection approaches:
1232
Analysis and research challenges. IEEE Access 7:182459-182476. 1231
Wazid M, Das AK, Rodrigues JJ, Shetty S, and Park Y. 2019b. IoMT malware detection approaches:
1232
Analysis and research challenges. IEEE Access 7:182459-182476. 1233
Weyrich M, and Ebert C. 2015. Reference architectures for the internet of things. IEEE Software 33:112-
1234
116. 1235
Wolfgang E. 2011. Introduction to artificial intelligence. Translated by Nathanael Black With illustrations
1236
by Florian Mast, Springer-Verlag London Limited. 1237
Wu F, Li X, Sangaiah AK, Xu L, Kumari S, Wu L, and Shen J. 2018. A lightweight and robust two-factor
1238
authentication scheme for personalized healthcare systems using wireless medical sensor
1239
networks. Future Generation Computer Systems 82:727-737. 1240
h
//d i
/10 1016/j f
2017 08 042 1241
Xiao L, Wan X, Lu X, Zhang Y, and Wu D. Manuscript to be reviewed 2018. IoT security techniques based on machine learning: How
1242
do IoT devices use AI to enhance security? IEEE Signal Processing Magazine 35:41-49. y
g
g
g
1243
Xing K, Srinivasan SSR, Jose M, Li J, and Cheng X. 2010. Attacks and countermeasures in sensor networks:
1244
a survey. Network security: Springer, 251-272. 1243
Xing K, Srinivasan SSR, Jose M, Li J, and Cheng X. 2010. Attacks and countermeasures in sensor networks:
1244
a survey. Network security: Springer, 251-272. 1245
Xu G, Lan Y, Zhou W, Huang C, Li W, Zhang W, Zhang G, Ng EYK, Cheng Y, Peng Y, and Che W. 2019. An
1246
IoT-Based Framework of Webvr Visualization for Medical Big Data in Connected Health. IEEE
1247
Access 7:173866-173874. 10.1109/ACCESS.2019.2957149 PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Computer Science Manuscript to be reviewed Manuscript to be reviewed Figure 1 Figure 1: The keywords used in the Research Information Template (RIT). Manuscript to be reviewed 1248
Xu J, Wei L, Wu W, Wang A, Zhang Y, and Zhou F. 2020. Privacy-preserving data integrity verification by
1249
using lightweight streaming authenticated data structures for healthcare cyber–physical system. 1250
Future Generation Computer Systems 108:1287-1296. 1251
https://doi.org/10.1016/j.future.2018.04.018
1252
Yaacoub J-PA, Noura M, Noura HN, Salman O, Yaacoub E, Couturier R, and Chehab A. 2020. Securing
1253
internet of medical things systems: Limitations, issues and recommendations. Future Generation
1254
Computer Systems 105:581-606. https://doi.org/10.1016/j.future.2019.12.028
1255
Yang Y, Zheng X, Guo W, Liu X, and Chang V. 2018. Privacy-preserving fusion of IoT and big data for e-
1256
health. Future Generation Computer Systems 86:1437-1455. 1257
https://doi.org/10.1016/j.future.2018.01.003
1258
Yeh K-H. 2016. BSNCare+: A robust IoT-oriented healthcare system with non-repudiation transactions. 1259
Applied Sciences 6:418. 1260
Zhang M, Raghunathan A, and Jha NK. 2013. MedMon: Securing medical devices through wireless
1261
monitoring and anomaly detection. IEEE Transactions on Biomedical circuits and Systems 7:871-
1262
881. 1263
Zhang Y, Gravina R, Lu H, Villari M, and Fortino G. 2018. PEA: Parallel electrocardiogram-based
1264
authentication for smart healthcare systems. Journal of Network and Computer Applications
1265
117:10-16. 1266
1267
1268
1269
1270 1248
Xu J, Wei L, Wu W, Wang A, Zhang Y, and Zhou F. 2020. Privacy-preserving data integrity verification by
1249
using lightweight streaming authenticated data structures for healthcare cyber–physical system. 1250
Future Generation Computer Systems 108:1287-1296. 1251
https://doi.org/10.1016/j.future.2018.04.018 1252
Yaacoub J-PA, Noura M, Noura HN, Salman O, Yaacoub E, Couturier R, and Chehab A. 2020. Securing
1253
internet of medical things systems: Limitations, issues and recommendations. Future Generation
1254
Computer Systems 105:581-606. https://doi.org/10.1016/j.future.2019.12.028 1255
Yang Y, Zheng X, Guo W, Liu X, and Chang V. 2018. Privacy-preserving fusion of IoT and big data for e-
1256
health. Future Generation Computer Systems 86:1437-1455. 1257
h
//d i
/10 1016/j f
2018 01 003 1258
Yeh K-H. 2016. BSNCare+: A robust IoT-oriented healthcare system with non-repudiation transactions. 1259
Applied Sciences 6:418. 1260
Zhang M, Raghunathan A, and Jha NK. 2013. MedMon: Securing medical devices through wireless
1261
monitoring and anomaly detection. IEEE Transactions on Biomedical circuits and Systems 7:871-
1262
881. 1263
Zhang Y, Gravina R, Lu H, Villari M, and Fortino G. 2018. PEA: Parallel electrocardiogram-based
1264
authentication for smart healthcare systems. Journal of Network and Computer Applications
1265
117:10-16. PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Computer Science Manuscript to be reviewed Figure 1: The keywords used in the Research Information Template (RIT). The entire Mind Map show The keywords used in the Research Information Template (RIT). The rectangle box at the middle represent the main keywords, then the square boxes
represent the derived similar words from the main words. The oval shaped keywords are
dervied from their previous sequare box keywords PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Figure 2 Figure 2: The search strategy used for selecting the research papers based on the
PRISMA guideline. The flow chart represents the procedure of searching in different databases using PRISMA
guideline. It starts from top to bottom showing each step of paper selection and fileteration . PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Figure 3 Figure 3: Geographical distribution of the papers. Computer Science Computer Science Manuscript to be reviewed Manuscript to be reviewed Manuscript to be reviewed PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Figure 4 Figure 4: Distribution of the papers by year. Figure 3: Geographical distribution of the papers. The pie chart shows the percentage of the papers by each country PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Computer Science Manuscript to be reviewed Manuscript to be reviewed Figure 4: Distribution of the papers by year. Each Blue Bar represent the number of papers published in each year and orange line shows
the percentage of the included reviewed papers in each single year PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Computer Science Manuscript to be reviewed Figure 5 Figure 5: The type of analyzed papers used in the current research. The pie chart shows the percentage of the analyzed papers in each catgory of Journal,
Conference, and Book chapter The pie chart shows the percentage of the analyzed papers in each catgory of Journal,
Conference, and Book chapter e pie chart shows the percentage of the analyzed papers in each catgory of Journal,
nference, and Book chapter
PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Computer Science Manuscript to be reviewed Manuscript to be reviewed Computer Science Computer Science Manuscript to be reviewed Manuscript to be reviewed Figure 6 Figure 6: Distribution of the papers according to the publishers. the pie chart line chart shows the percentage of the analyzed papers which are published by
each publisher PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Figure 7 Figure 7: Papers distribution by the direction and problem-solving domain. The pie chart show the direction of studies. The percentage of the papers for each direction
of study is given in the chart PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Computer Science Manuscript to be reviewed Manuscript to be reviewed Figure 8 Figure 8. The type of data used by the researchers to conduct their research. The pie chart shows the percentage of the papers in terms of the type of the data used for
their analysis PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Computer Science Manuscript to be reviewed Manuscript to be reviewed Figure 9 Figure 9: Software and tools used by the studies. Figure 9: Software and tools used by the studies. The pie chart shows the percentage of different tools and software which are used by the
reviewed studies PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Computer Science Manuscript to be reviewed Computer Science Computer Science Manuscript to be reviewed Manuscript to be reviewed Figure 10: Devices and hardware tools used in the studies. Figure 10: Devices and hardware tools used in the studies. The bar chart shows the number of hardware tools used by the analyzed studies . The bar chart shows the number of hardware tools used by the analyzed studies . PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Figure 11 Figure 11: Analysis of the studies in terms of (a)complexity and (b) real time analysis
The pie chart ( a ) shows the percentage of the papers in terms of complexity {lightweight,
heavy, low complex} The pie chart (b) shows the percentage of the analyzed papers based Figure 11: Analysis of the studies in terms of (a)complexity and (b) real time analysis Figure 11: Analysis of the studies in terms of (a)complexity and (b) real time analysis The pie chart ( a ) shows the percentage of the papers in terms of complexity {lightweight,
heavy, low complex} The pie chart (b) shows the percentage of the analyzed papers based
on their real time feature {real time, not real time (heavy), low time complex)} PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Manuscript to be reviewed Manuscript to be reviewed Computer Science Manuscript to be reviewed Table 1(on next page) Figure 12 Figure 12: Details of the studies in terms of placement. the chart shows the percentage of the papers based on their placement the chart shows the percentage of the papers based on their placement PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Table 1(on next page) Table 1(on next page) Table 1: The types and characteristics of medical devices used in the IoMT system. Each row represent different type of medical device and each column represents
characteristic of that device PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Manuscript to be reviewed Table 2(on next page) Table 2(on next page) Manuscript to be reviewed 1
Table 1:
2
The types and characteristics of medical devices used in the IoMT system. Device type
Placement
Example
Risk
Value
Ref. Implantable
H Within the human tissues
deep brain implants, heart
pacemaker and insulin
pump
High
(Santagati et al. 2020)
Wearable
On the human body
smart watches, fitness
devices
Low
(Tseng et al. 2019)
Ambient
Outside the human body
elderly
monitoring
devices in smart home
Low
(Pandey
&
Litoriya 2020)
Stationary
Inside hospitals
medical image processing
devices of MRI and CT-
Scan
Low
(Nanayakkara
et al. 2019; Xu
et al. 2019)
3 1
Table 1: PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Table 2(on next page) Table 2: Attacks on different the IoMT layers with their respective impact on security
requirements Each row reprsenst different type of attack and the rows show their targeted layer, Impact,
and Refrenece PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Manuscript to be reviewed Table 3(on next page) Manuscript to be reviewed 1
Table 2:
2
Attacks on different the IoMT layers with their respective impact on security requirements. Attacks
Impact
Ref. Targeted layer: Server/Database layer
Malware attack
Integrity, Availability
(Wazid et al. 2019)
Ransomware attack
Integrity, Availability
(Fernandez Maimo et al. 2019)
SQL injection
All
(Li et al. 2019, Gupta et al. 2020)
Social Engineering (Reverse
Engineering, Shoulder-surfing)
All
(Yaacoub et al. 2020) (Patel 2020) (He et al. 2018)
Brute Force
Confidentiality, Integrity
(Stiawan et al. 2019)
Adversarial Machine Learning attacks
(Causative (Poisoning and Evasion
attacks), Exploratory)
Confidentiality, Integrity
(Ibitoye, Shafiq and Matrawy 2019), (Mozaffari-Kermani
et al. 2014, Mosenia and Jha 2016), (Chakraborty et al. 2018)
Targeted layer: Network layer
Denial of Service (DoS) and
Distributed DoS (DDoS)
All
(Gupta et al. 2020, Yaacoub et al. 2020) , (Al Shorman,
Faris and Aljarah 2019)
Man in the Middle (MIM) attack
Confidentiality, Integrity
(Yaacoub et al. 2020)
Eavesdropping attack
Confidentiality, Non-repudiation, Privacy
(Gupta et al. 2020) (McMahon et al. 2017)
Replay attack
Confidentiality, Integrity
(Spiekermann 2015, Yaacoub et al. 2020)
botnet attack
Availability, Confidentiality
(Bahşi, Nõmm and La Torre 2018)
Mirai attack
Availability, Confidentiality
Jamming
Availability
(Sun, Lo and Lo 2019)
flooding
Availability
(Gupta et al. 2020, Shafiq et al. 2020, Yaacoub et al. 2020)
Packet Analysis attacks
Integrity, Confidentiality, Non-
repudiation, Privacy
(Tseng, Wu and Lai 2019)
Targeted layer: Device/sensor layer
Physical Sensor/ Node tampering
All
(Xing et al. 2010)
False data Injection
Integrity, Confidentiality, Non-
repudiation, Privacy
(Rahman and Mohsenian-Rad 2012, Bostami, Ahmed and
Choudhury 2019, Farroha 2019, Newaz et al. 2020)
Resource Depletion Attacks (Battery
drain, Sleep deprivation, Buffer
overflow)
Availability
(Hei et al. 2010), (Mosenia and Jha 2016) , (Gupta et al. 2020)
Side-channel
Confidentiality, Non-repudiation, Privacy
(Zhang, Raghunathan and Jha 2013, Newaz et al. 2020)
Hardware Trojan
All
(Qu and Yuan 2014, Mosenia and Jha 2016)
Eavesdropping
Confidentiality, Non-repudiation, Privacy
(Gupta et al. 2020) (McMahon et al. 2017)
3 1
Table 2: 3 PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Table 3(on next page) Table 3: The evaluation metrics used for ML techniques. Each row represents a metric and the columns show their description, and formula Each row represents a metric and the columns show their description, and formula PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Manuscript to be reviewed Manuscript to be reviewed Table 4(on next page) Manuscript to be reviewed 1
Table 3:
2
The evaluation metrics used for ML techniques. Metric
Description
Formula
Accuracy
Determines the performance of the model in recognizing all classes, respectively
𝐴𝑐𝑐=
𝑇𝑃+ 𝑇𝑁
𝑁
Sensitivity
(Recall)
Measures the completeness, which is the percentage of positive predicted samples
to the positive samples in dataset is depicted. 𝑆𝑒𝑛𝑠𝑖𝑡𝑖𝑣𝑖𝑡𝑦=
𝑇𝑃
𝑇𝑃+ 𝐹𝑁
Specificity
(Precision)
Shows the exactness, in which the percentage of correctly positive predictive
samples to all positive predictive samples by the model are calculated. 𝑆𝑝𝑒𝑐𝑖𝑓𝑖𝑐𝑖𝑡𝑦=
𝑇𝑃
𝑇𝑃+ 𝐹𝑃
False Positive
Rate (FPR)
Measures the number of those normal network behaviors which are calculated as
attack. 𝐹𝑃𝑅 = 𝐹𝑃/( 𝐹𝑃+ 𝑇𝑁)
Performance
overhead
It is the calculation of any combination of (memory, CPU, energy) overhead taken
by the ML techniques to perform a task. Big(o)notation for time and
space complexity and Energy
unit for energy usage
Where TP is True Positive, TN is True Negative, N is total number of samples, FP is False Negative FP is False Positive
3 1
Table 3: 4 PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Table 4(on next page) Table 4: Comparison between this survey and other related surveys. Each row show different related study and columns show their features PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Manuscript to be reviewed Table 5(on next page) Manuscript to be reviewed Manuscript to be reviewed 1
Table 4:
2
Comparison between this survey and other related surveys. Year
Ref. IoT
domain
Architecture
Threats
ML
methods
Big data
ML for
IoMT
security
Systematic
analysis
2018
(Cui et al. 2018)
Generic
NA
NA
Discussed
NA
NA
NA
2020
(Tahsien et
al. 2020)
Generic
IoT
architecture
IoT attacks
Discussed
NA
NA
NA
2020
(Hussain et
al. 2020)
Generic
NA
IoT attacks
Discussed
Big data at
cloud
NA
NA
2019
(Sun et al. 2019)
IoMT
IoMT
architecture
IoMT
Security
requirement
NA
NA
Partially
discussed
NA
2020
(Newaz et al. 2020)
IoMT
IoMT
architecture
IoMT attacks
NA
NA
Partially
discussed
NA
2020
(Yaacoub et
al. 2020)
IoMT
IoMT
architecture
IoT attacks
NA
NA
NA
NA
-
This study
IoMT
IoMT
architecture
IoMT
Attacks
Discussed
Discussed
Discussed
Comprehensive
and Systematic
Review
3 1
Table 4: 3 PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed
Computer Science Manuscript to be reviewed
Computer Science Table 5(on next page) Table 5: The searching queries and results achieved from five different databases. Table 5: The searching queries and results achieved from five different databases. Table 5: The searching queries and results achieved from five different databases. Each row shows different queries used for all databases and the columns show their results PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) 2
The searching queries and results achieved from five different databases. esults achieved from five different databases. Computer Science Manuscript to be reviewed Table 6(on next page) Table 6(on next page) Manuscript to be reviewed 1
Table 5: 2
Th
hi
i
d
lt
hi
d f
fi
diff
t d t b
Searching texts versus databases
IEEE
Xplore
Springer
Link
Scopus
Science
Direct
Web of
Science
Machine Learning AND Medical Internet of Things AND Security
89
3431
63
2459
33
Machine Learning AND Medical Internet of Things AND Privacy
37
1093
43
1494
31
Machine Learning AND Medical Internet of Things AND Intrusion
6
336
2
445
3
Machine Learning AND Medical Internet of Things AND Attack
13
1225
2
1203
18
Machine Learning AND IoMT AND Security
9
81
3
929
5
Machine Learning AND IoMT AND Privacy
8
44
3
78
6
Machine Learning AND IoMT AND Intrusion
0
8
1
21
1
Machine Learning AND IoMT AND Attack
0
33
64
5
Machine Learning AND Medical Cyber Physical system AND Security
30
481
5
1038
11
Machine Learning AND Medical Cyber Physical system AND Privacy
8
429
3
693
4
Machine Learning AND Medical Cyber Physical system AND
Intrusion
6
321
3
301
4
Machine Learning AND Medical Cyber Physical system AND Attack
9
494
0
699
11
Machine Learning AND Smart healthcare AND Security
57
1971
12
2134
52
Machine Learning AND Smart healthcare AND Privacy
25
1012
3
1901
36
Machine Learning AND Smart healthcare AND Intrusion
7
334
1
431
21
Machine Learning AND Smart healthcare AND Attack
20
1031
3
1209
20
Total including duplicates
324
12324
147
15099
261 4 PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Table 6(on next page) Table 6: Selection criteria of the papers at final stage Table 6: Selection criteria of the papers at final stage
each row represent a criadera for selecting the papers and the coloumn shows the responc each row represent a criadera for selecting the papers and the coloumn shows the responce PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Manuscript to be reviewed Table 7(on next page) Manuscript to be reviewed 1
Table 6:
2
Selection criteria of the papers at final stage
Criteria#
Questions
Answer
1
Does the paper relevant to the topic? Y/N
2
Does the work propose a machine learning related solution and
method to solve a problem in the IoMT security and/or
privacy? Y/N
3
Is the paper published in scholarly journals, conferences,
books? Y/N
3 1
Table 6:
2
Selection criteria of the papers at final stage
Criteria#
Questions
Answer
1
Does the paper relevant to the topic? Y/N
2
Does the work propose a machine learning related solution and
method to solve a problem in the IoMT security and/or
privacy? Y/N
3
Is the paper published in scholarly journals, conferences,
books? Y/N
3 3 PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Table 7(on next page) Table 7: Classification of papers based on ML category, medical device, and the IoMT
layer. The table shows a matrix represntation of the paper's classification to different categories The table shows a matrix represntation of the paper's classification to different categories PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Manuscript to be reviewed Table 8(on next page) Table 8(on next page) Computer Science Manuscript to be reviewed Table 7:
Classification of papers based on ML category, medical device, and the IoMT layer. ML category - medical device category
IoMT layer- Ref
Supervised ML - Implantable, wearable
Sensor layer- { (Haque, Rahman and Aziz 2015) ,(Abdaoui et al. 2020)
(Rathore et al. 2018), (Khan et al. 2017), (Gao and Thamilarasu 2017), (Newaz
et al. 2019), (Ben Amor, Lahyani and Jmaiel 2020), (Mohamed, Meddeb-
Makhlouf and Fakhfakh 2019), (Salem et al. 2014), (Hau and Lupu 2019),
(Nagdeo and Mahapatro 2019), (Verner and Butvinik 2017) }
All layers-(Rathore et al. 2018b)
Supervised ML- Wearable, smart watches,
smart fitness
Device layer- {(Pirbhulal et al. 2019), (Mawgoud et al. 2019), (Barros et al. 2019), (Zhang et al. 2018), (Shang & Wu 2019), (Musale et al. 2019), (Vhaduri
& Poellabauer 2019), (Mohsen et al. 2019), (Rathore et al. 2018a) }
Network layer- {(Begli, Derakhshan and Karimipour 2019), (Itten and
Vadakkumcheril 2016), (Schneble and Thamilarasu 2019), (Odesile and
Thamilarasu 2017), (RM et al. 2020), (Asfaw et al. 2010), (Alrashdi et al. 2019), (Wazid et al. 2019), (Fernandez Maimo et al. 2019)}
Cloud layer- {(Punithavathi et al. 2019), (Landau et al. 2020)}
Unsupervised ML-Implantable, wearable
Sensor layer- {(Ben Amor, Lahyani and Jmaiel 2020),(Sehatbakhsh et al. 2018), (Zhang, Raghunathan and Jha 2013), (Verner and Butvinik 2017)}
Unsupervised ML-Wearable
Device layer- {(Zhang et al. 2018), (Shang & Wu 2019), (Musale et al. 2019),
(Vhaduri & Poellabauer 2019), (Mohsin et al. 2019)}
Network layer- {(Schneble and Thamilarasu 2019), (He et al. 2019),(RM et al. 2020), (Thamilarasu 2016), Fernandez Maimo et al. 2019)}
Unsupervised ML-ICD, Programmer
Device layer- (Kintzlinger et al. 2020)
Deep learning-Wearable
Device layer-{ (RM et al. 2020), (Manimurugan et al. 2020), (Shakeel et al. 2018), (Rathore et al. 2019), (Mohsin et al. 2019) }
Big data-Wearable
Device layer-(Zhang et al. 2018) PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 2
Table 8:
3
Details of published studies on anomaly and attack detection to the sensors/medical devices
Ref. Methods
Detection type
Good features
Limitations
Tools
Dataset info. (Haque, Rahman
and Aziz 2015)
SMO
Anomaly detection
- high detection rate
- low FPR
- high computation
overhead
Weka
-10 real
datasets
-(MIMIC)data
(Abdaoui et al. 2020)
Deep
Learning
Anomaly based false
alarm detection
-real time
-high accuracy
- high computation
overhead
- high FPR
Tensor flow
and Keras in
Python
John
Radcliffe
Hospital data
(Rathore et al. 2018)
neural
network-
based MLP
Anomaly based false
alarm detection
-real time
-energy efficient
-high accuracy
-reliable
- high memory
requirement due to
training overhead
NI myRIO
FPGA-based
MLP
UCI
Diabetic
dataset
(Khan et al. 2017)
DWT and
Marcov
model
Anomaly based false
data detection
-high detection rate
-high TNR
-real time
- detection rate decreases
when there is too much
attack
- high FNR and FPR
MATLAB
The ECG
dataset from
MIT-
PHYSIOBAN
K
(Gao and
Thamilarasu
2017)
Decision
tree, SVM
and K-
means
Anomaly-based attack
detection
- high accuracy
- low FPR
- low training time
- low prediction time
- no validation
- memory, battery usage is
not considered
- fails in detecting insider
attack
Castalia
Simulation
data
(Sehatbakhsh et
al. 2018)
(K-S test)
on external
hardware
device
malware Anomaly
detection
-high TPR
-low FPR
-low detection latency
-no overhead on the
medical device
- external device needs
maintenance and the
device itself could be
hacked (stolen or lost)
Open Syringe
Pump
Arduino UNO,
Nios-II,
OlimexA13,
and TS-7250
Testbed data
(Zhang,
Raghunathan and
Jha 2013)
a model is
embedded
on an
external
device
multi-layered anomaly
detection. - zero overhead on
battery
- real time
- multiple attacks
detection
- hybrid detection
-protecting only integrity
- external device needs
maintenance and it could
be hacked (stolen or lost)
- lack of behavioral
anomaly detection
experiment
USRP
Glucose
monitoring
and insulin
delivery
systems
Testbed data
(Newaz et al. 2019)
ANN, DT,
RF, and k-
NN
Anomaly detection
Uses medical device
data
-high accuracy
-high F1
-No overhead on the
sensors
- uses body
functioning data
- high training overhead
- performance overhead
not calculated. MATLAB
A set of heath
dataset from
different
sources
(Ben Amor,
Lahyani and
Jmaiel 2020)
PCA and
Correlation
Coefficient
Anomaly based faulty
sensor data detection. Table 8(on next page) Table 8: Details of published studies on anomaly and attack detection to the
sensors/medical devices each row represent each paper under category anomaly and attack detection to the
sensors/medical devices and the columns show their characteristics that are used for
evaluating them PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed - real time
- lightweight
- improved accuracy
- improved FPR
-energy, CPU usage is not
considered
-lacks attack detection at
server and transmission
AUDITmodul
e
Java and R
languages
MIMIC
database
(Mohamed,
Meddeb-
Makhlouf and
Fakhfakh 2019)
Statistical
signal
amplitude
calculation
Anomaly based
intrusion cancellation
- using more than one
type of Sensor type
- high TPR
- not lightweight
-not real time
-high performance
overhead
MATLAB
real
medical ECG
and EMG
datasets
(Salem et al. 2014)
Classifiers
(SVM, RF,
K-NN,
Decision
Trees) and
Regression
Anomaly detection for
sensor physiological
data
- low detection time
- high TPR
- low FNR
-not lightweight
-performance overhead not
calculated
Weka
MIMIC
dataset
(Hau and Lupu
Time series
Anomaly based false
-High detection for
-long detection time
Not given
MIMIC PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Computer Science Manuscript to be reviewed Table 9(on next page) Manuscript to be reviewed 2019)
approach
temporal-
attribute
correlations
data injection attack
detection
single sensor
-acceptable detection
for moderate sensors
-disable when many
sensors increase
(collusion)
dataset
(Nagdeo and
Mahapatro 2019)
combined
ANN with
Ensemble
LinReg
Classification of
anomalous and faulty
sensor physiological
data
- high TPR
- low FPR
Not real-time
Not lightweight
Unknown dataset source
Performance overhead not
calculated
Weka
ECG dataset
(Verner and
Butvinik 2017)
Otsu’s
Thresholdin
g, and
Linear SVM
sensor data
modification detection
- high precision
- high recall
- can be used for other
IoT domains
-not real-time
-not lightweight
-Performance overhead not
calculated
MATLAB
(JDRF)
(CGM)
Clinical Trial
dataset
(Kintzlinger et al. 2020)
statistical
and One
class SVM
Rule and knowledge
based anomaly attack
detection
- high TPR
- low FPR
- high accuracy
- real time
-layers III-V is useless
- heavy training overhead
- performance overhead
not calculated
- learns from only benign
data
Not given
Self-created
clinical data
(Rathore et al. 2019)
Deep
Learning
classifier
Attack prediction
-classify different
attacks
-reduced training time
-not lightweight
-accuracy not given
Keras with
Theano in
Python
Parkin
tremo
datase
Physi 4 PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 1
Table 9:
2
The summary of the studies reported on authentication and access control. Ref. Methods
Good Features
Gaps and limitation
tools
Data info
(Rathore et al. 2018b)
Legendre
approximation
and MLP
-High testing
accuracy
-High computational overhead for IMD. -Not able to detect unknown attacks. MATLAB
Keras
Theano
ECG-ID dataset
(Pirbhulal et
al. 2019)
MLP
-Efficient
-Light weight
-Evaluation not given. -Lacks validation metrics. -MLP is memory intensive. Not given
Not given
(Mawgoud et
al. 2019)
SVM and trust
management
-Low resource
consumption
-Lower detection
time
-Performance overhead not calculated. -Lacks measuring the cryptography security
strength of the algorithm. Not given
Not available
(Barros et al. 2019)
MLP, RF,
SVM and
Naïve bayes
-Low complexity
-High accuracy
- Performance overhead not calculated
-High Training overhead
-Reduced accuracy due to feature selection
-Not easy to implement with different sensors
Weka
Stress
Recognition in
Automobile
Drivers
database
(Zhang et al. 2018)
2DPCA, LDA,
and
MapReduce
-Improved accuracy
-Improved
efficiency
-Performance overhead not calculated. -High training overhead
- Not suitable for tiny IoT devices
Hadoop
MIT-BIH
Database
(Shang & Wu
2019)
LOF model
-Improved accuracy
in acceptance and
rejection
-Not accurate when user’s behavior not stable. -Different performance on different brands of
smartwatch
-Sensors should be attached to body tightly. -Performance overhead not calculated
Samsung
smartwatchTi
zen OS 3.0
Self-created
(Musale et al. 2019)
Statistical
filers, RF,
KNN,
and MLP
-Lightweight
-Higher accuracy
-User-friendly
-Easily deployable
- performance overhead not calculated. - No attack model is considered for gait
authentication. - Environment can affect its accuracy. - Authentication fails if the user is far from
smart home. - Its accuracy decreases in the case of
increasing the users
Motorola
(smartphone)
python
Self-created
(Vhaduri &
Poellabauer
2019)
A combination
of filters
((KS)-test, PC,
SD based
filter ) and
SVM
-Use of hybrid
biometric. -High accuracy
-Low error rate
-Degradation of Non-Sedentary performance
for highly active periods. -Need for retraining. -Cannot detect online attacks
-performance overhead not calculated. MATLAB
NetHealth
Study Dataset
(Mohsen et al. 2019)
Cryptography,
convolutional
Neural
network
(CNN)
-End to end security
-Strong against
multiple attacks
-Lightweight
- Real time
-Communication overhead is still high for
medical devices. -High training overhead. - Performance overhead not calculated. -Some attacks may still occur in that 30 min of
monitoring gap ex; sensor may be relocated and
returned
Not available
Not available
(Punithavathi
et al. Table 9(on next page) Table 9: The summary of the studies reported on authentication and access control. each row represent each paper under category authentication and access control and the
columns show their characteristics that are used for evaluating them PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 2019)
a lightweight
random
projection
technique
-Low complexity
-Lightweight
-High recognition
accuracy
-Rejection rate not calculated. -memory, energy usage is not calculated. -Missing attack validation
OpenCV 3.6
in Python 3.4. DB1 and DB2
of FVC2002
and FVC2004
(Rathore et al. 2018a)
Dynamic
Time Warping
(DTW)
-Lightweight
-Efficient
-High accuracy
-Less complex
-Missing validation against attacks
-Acceptance and Rejection rate not calculated. -Heavy for implantable devices
LabVIEW
MathScript
RT Module
ECG-ID dataset
(Mohsin et al. 2019)
PSO,
Blockchain,
AES
-Higher secure
transmission
-Strong against
spoofing and brute-
-Key distribution in AES adds extra load on the
channels. -Block chain is a heavy and resource extensive
Not given
MMCBNU_600
0 database 1
Table 9: PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) 3
force attacks
-High accuracy
Manuscript to be reviewed
Computer Science Computer Science Manuscript to be reviewed Manuscript to be reviewed Table 10(on next page) Computer Science Manuscript to be reviewed 3 PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed 1
2
Table 10:
3
Summary of the studies reported on intrusion and malware detection. Ref. Method
Type of Intrusion
detection
Good features
Gaps and Limitations
Tools
(software&
hardware)
Datasets
(Begli,
Derakhshan and
Karimipour
2019)
SVM
Anomaly and
signature-based
IDS
-High detection
accuracy
-Satisfied detection
time
- High memory
overhead
- Low detection rate in
misuse analysis
-Outdated dataset
R program
NSL-KDD
(Itten and
Vadakkumcheril
2016)
a multivariate
correlation analysis
Hybrid Anomaly
and rule based
IDS
- Improved
accuracy. - Detecting known
and unknown
attacks. - High learning
overhead
- Unclear
implementation
- Validation process
not given
Not given
Not given
(Schneble and
Thamilarasu
2019)
Federated Learning
Anomaly based
false data
injection, data
modification, DoS
IDS
-Multiple attack
detection
-High accuracy
- Low FPR
- Flexible
- Reduced accuracy
and increased FPR in
some cases
- Weak against
adversarial attacks
- Sci-kit
Learn’s on
Raspberry
Pi’s. - MATLAB
- MIMIC dataset
from PhysioNet
- ECG
(Odesile and
Thamilarasu
2017)
Hierarchical and
distributed classifiers
(NBC, KNN, SVM,
RF and DT)
Anomaly based
IDS
- High accuracy
- Low energy
consumption
- High training time
for some methods
- High FPR
Castalia
WBAN
Self-created
simulated data
(Schneble and
Thamilarasu
2019)
feature selection
(Laplacian scoring)
Signature based
intrusion
detection. - Reduced the
detection time. - Reduced
performance
overhead
- Accuracy reduced
upon selecting more
features. - Lacks detail of
selected features. Not given
MIT-BIH
Arrhythmia
dataset
(He et al. 2019)
SAE is used for
feature selection
Signature based
intrusion
detection. HIDS
- Improved
accuracy and
detection overhead
- High training
overhead
- Not lightweight
Not given
real CHS Source
not given
(RM et al. 2020)
Feature selection
using PCA and
GWO with DL
classifier
Signature based
IDS
NIDS
-high accuracy
low training and
testing time
- High Memory and
CPU overhead. - Limited to only IP
based devices. Not given
NSL-KDD
(Thamilarasu
2016)
multi-objective
genetic algorithm
(GA)
-Signature based
IDS
-HIDS
-F selection
reduced the
complexity
- Not lightweight
- Detection time and
training time not given. MATLAB
Self-created by
simulation
(Asfaw et al. 2010)
Data mining-based
Association rule
mining
-Anomaly based
HIDS
-Ability to detect
anomalous
activity. - Lacks model
evaluation
- Using many
resources
-J2ME
-Java Servlets
-MySQL
Server
-Palm PDA
Self-created
(Manimurugan
et al. 2020)
Deep Belief Network
(DBN)
-Anomaly based
NIDS. -High accuracy
-High precision,
F1, and recall. - FPR and performance
overhead neglected. Manuscript to be reviewed
Computer Science Manuscript to be reviewed
Computer Science Table 10(on next page) Table 10: Summary of the studies reported on intrusion and malware detection. Table 10: Summary of the studies reported on intrusion and malware detection. each row represent each paper under category intrusion and malware detection, the columns
show their characteristics that are used for evaluating them each row represent each paper under category intrusion and malware detection, the columns
show their characteristics that are used for evaluating them PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed - -High training
overhead
MATLAB
CICIDS2017
dataset PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021) Manuscript to be reviewed (Alrashdi et al. 2019)
Ensemble
an online sequential
extreme learning
machine (EOS-
ELM)
-Anomaly based
NIDS. - Lower latency
compared with
cloud-based. - Reduced attack
detection time
-Memory and CPU
usage are not
considered
-Python
(scikit-learn,
Tensorflow,
Keras,
Numpy,
HDF5)
NSL-KDD
dataset
(Wazid et al. 2019)
Machine Learning,
Data mining,
Blockchain
-Signature and
Anomaly based
Malware
detection. Discussed current
IoMT malware
detection
Most papers are
generic IoT especially
Smart phone-based
solutions. Not given
Not given
(Fernandez
Maimo et al. 2019)
NFV / SDN, OC-
SVM , and Naïve
Bayes
-Anomaly and
signature-based
Ransomware
detection. - Real time
- Can detect recent
malware attacks. - Short detection
time
- Not lightweight
- performance
overhead not
calculated
- OpenICE
- OpenStack
-
OpenDaylight
- Python
language
- Scikit-learn
v0.20.0
Self-created real
testbed
dataset
(Shakeel et al. 2018)
Deep-Q-Network
(LDQN)
-Signature based
malware
detection. - High detection
rate
- Low error
- Low energy
consumption
- FPR, Memory usage
are not considered. - Training overhead is
high
NS2
simulator
Simulated data
(Landau et al. 2020)
Different ML
techniques
-Privacy attack
detection. -Improved
performance
-Larger datasets
used
- Still low accuracy
- high error rare
Not given
rsEEG data
4 4 5 5 PeerJ Comput. Sci. reviewing PDF | (CS-2020:10:53665:1:1:NEW 19 Jan 2021)
|
https://openalex.org/W2113371027
|
https://bmcsurg.biomedcentral.com/counter/pdf/10.1186/1471-2482-11-9
|
English
| null |
Enhanced recovery in colorectal surgery: a multicentre study
|
BMC surgery
| 2,011
|
cc-by
| 6,054
|
RESEARCH ARTICLE Open Access © 2011 Ramírez et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons
Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in
any medium, provided the original work is properly cited. Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Enhanced recovery in colorectal surgery:
a multicentre study José M Ramírez1, Juan A Blasco2, José V Roig3, Sergio Maeso-Martínez2*, José E Casal4, Fernando Esteban5,
Daniel Callejo Lic2 and for Spanish working group on fast track surgery Abstract Background: Major colorectal surgery usually requires a hospital stay of more than 12 days. Inadequate pain
management, intestinal dysfunction and immobilisation are the main factors associated with delay in recovery. The
present work assesses the short and medium term results achieved by an enhanced recovery program based on
previously published protocols. Methods: This prospective study, performed at 12 Spanish hospitals in 2008 and 2009, involved 300 patients. All
patients underwent elective colorectal resection for cancer following an enhanced recovery program. The main
elements of this program were: preoperative advice, no colon preparation, provision of carbohydrate-rich drinks
one day prior and on the morning of surgery, goal directed fluid administration, body temperature control during
surgery, avoiding drainages and nasogastric tubes, early mobilisation, and the taking of oral fluids in the early
postoperative period. Perioperative morbidity and mortality data were collected and the length of hospital stay and
protocol compliance recorded. Results: The median age of the patients was 68 years. Fifty-two % of the patients were women. The distribution of
patients by ASA class was: I 10%, II 50% and III 40%. Sixty-four % of interventions were laparoscopic; 15% required
conversion to laparotomy. The majority of patients underwent sigmoidectomy or right hemicolectomy. The overall
compliance to protocol was approximately 65%, but varied widely in its different components. The median length
of postoperative hospital stay was 6 days. Some 3% of patients were readmitted to hospital after discharge; some
7% required repeat surgery during their initial hospitalisation or after readmission. The most common
complications were surgical (24%), followed by septic (11%) or other medical complications (10%). Three patients
(1%) died during follow-up. Some 31% of patients suffered symptoms that delayed their discharge, the most
common being vomiting or nausea (12%), dyspnoea (7%) and fever (5%). Conclusion: The following of this enhanced recovery program posed no risk to patients in terms of morbidity,
mortality and shortened the length of their hospital stay. Overall compliance to protocol was 65%. The following of
this program was of benefit to patients and reduces costs by shortening the length of hospital stay. The
implantation of such programmes is therefore highly recommended. * Correspondence: smaemar@hotmail.com
2Health Technology Assessment, Agencia Laín Entralgo, Madrid, Spain
Full list of author information is available at the end of the article Background morbidity but to the conventional care protocol fol-
lowed. For decades this protocol has hardly been modi-
fied: it therefore does not take into account the
advances that have been made in the perioperative man-
agement of such patients. Major colorectal surgery, i.e., surgery that involves wide
resection of the colon and anastomosis, generally
involves a prolonged hospital stay - on average 12-14
days. A stay of one week is usually the minimum that
can be expected [1]. This prolonged occupation of a
hospital bed is not usually owed to problems of Inadequate pain management, intestinal dysfunction
and immobilisation have been recognised since at least
1997 as among the main factors delaying postoperative
recovery in patients subjected to major surgery [2]. This
led Kehlet et al [3] to propose a series of measures
designed to improve recovery following major colorectal Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Page 2 of 8 There were two meetings with at least two profes-
sionals of each centre, a surgeon and an anaesthetist. During these meetings discussions were held with
national and international experts who assisted the
group in the implementation of the program. These pro-
fessionals were the persons in charge to develop the
program in their centre and to starting the implementa-
tion of the protocol in two months after the second
work session. There were two annual reunions with the
group to supervise and improve the compliance of the
protocol. surgery in their well-known multimodal recovery pro-
gram. Currently, evidence-based multidisciplinary action
protocols of this kind, known initially as fast-track or
better called enhanced recovery programs (ERAS), are
not achieving the degree of implantation hoped for [3]. Some authors suggest this to be due to the organisa-
tional demands they make on surgeons, anaesthetists
and nursing staff, and to social, cultural and economic
realities [4]. surgery in their well-known multimodal recovery pro-
gram. Currently, evidence-based multidisciplinary action
protocols of this kind, known initially as fast-track or
better called enhanced recovery programs (ERAS), are
not achieving the degree of implantation hoped for [3]. Some authors suggest this to be due to the organisa-
tional demands they make on surgeons, anaesthetists
and nursing staff, and to social, cultural and economic
realities [4]. Study design
Thi This prospective study, which involved 300 patients, was
performed between 2008 and 2009. The ERAS used was
developed by the authors on the basis of published pro-
tocols [3,5,8-12]. The variables recorded included perio-
perative
mortality,
length
of
hospital
stay
and
compliance with the protocol. The study was presented to the Hospital Ethical Board
and accepted as this is an observational non-randomised
study based on the best available evidence. The research was conducted conformed to the Hel-
sinki Declaration and to local legislation. Patients gave
informed consent to participate in the study. This study
has been registered in the ISRCTN register with the
number ISRCTN16397735, you can also access to the
registration information by the URL: http://www.con-
trolled-trials.com/ISRCTN16397735. Inclusion and exclusion criteria A) Inclusion criteria. All patients had to be 18 years of
age or over and to be programmed for surgery for color-
ectal cancer without the need for a stoma or any further
surgical procedure. The present work analyses the short and medium
term results returned by an ERAS for colorectal surgery
based on previously published protocols, followed at 12
Spanish hospitals. B) Exclusion criteria: The need for emergency surgery,
an American Society of Anaesthesiologists (ASA) class
of IV, the need for a colostomy or ileostomy, the inabil-
ity to provide informed consent, diabetes, slow evacua-
tion previously documented by a digestive medicine
unit. Participating centres The twelve participating centres were chosen for their
organisational abilities and their experience and interest
in colorectal surgery and patient care. These centres,
distributed around Spain, ranged from large university
hospitals to medium and small area hospitals (the Clín-
ico from Zaragoza, La Paz, Clínico San Carlos and Gre-
gorio Marañón from Madrid, General from Valencia,
Mútua Terrassa, Do Meixoeiro from Vigo, Hospital
d’Igualada, La Mancha Centro from Alcázar de San
Juan, Universitario de Elche, Son Llatzer from Palma de
Mallorca and Fundación Calahorra hospitals) with the
collaboration of Health Technology Assessment Unit
from Agencia Laín Entralgo, Madrid. Background The results that can be achieved with ERAS - reduc-
tions in postoperative morbidity, average length of hos-
pital stay and the consumption of resources - are,
however, significant, and the general implantation of an
ERAS for patients who are to undergo colorectal surgery
is recommendable [5-8]. In some of the clinical practice
guides available at http://www.reducinglengthofstay.org/,
these programmes are considered to represent best clin-
ical practice according to current scientific evidence. Unfortunately, the results communicated regarding sur-
gery in an ERAS context have nearly all come from indi-
vidual institutions, although they include those of four
randomised clinical trial [5,9-11]. Recently the results of
an international study (five hospitals in different coun-
tries, four with no prior experience in ERAS) reporting
on patient follow-up, the degree of acceptance and the
degree of compliance with the protocol of a common
ERAS, have also become available [12]. The results of
these studies [5,9-12] suggest that just making a proto-
col available is insufficient for objectives to be achieved;
changes also need to be made to organisational strate-
gies and the medical professionals involved in pre, intra
and especially postoperative care require support, per-
haps via continuing education. Enhanced recovery program protocol The ERAS required that, during the preoperative period,
patients be given advice, that there be no preparation of
the colon and that patients receive four carbohydrate-
rich drinks (4 × 200 ml) one day prior to surgery plus
two further drinks (2 × 200 ml) on the morning of sur-
gery. During surgery, goal directed fluids were adminis-
tered using oesophageal Doppler monitoring, and
hypothermia and drainages avoided. After surgery, naso-
gastric tubes were not used, early mobilisation was prac-
tised, and oral fluids administered early. Table 1 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Page 3 of 8 summarises the protocol characteristics of the ERAS
f ll
d
postoperative vomiting and nausea, and the use of epi-
d
l
h
Table 1 Protocol characteristics of the followed enhanced recovery program
Time
Procedure
Preoperative
a. Provision of verbal and written information to patients regarding the ERAS. Collection of signed consent. b. Malnourished patients to receive hyperproteic supplement at least twice per day during the week before surgery. Day before surgery
a. No colon preparation. b. Normal food in the morning. Liquids on demand during the evening. Four bricks of carbohydrate-rich Nutricia Preop®
to be taken during the evening (total: 800 ml). c. Prophylaxis for pulmonary thromboembolism following normal practice. d. Antibiotic prophylaxis following normal practice. Day of surgery (before
surgery)
a. Two hours before surgery: provision of two bricks of Nutricia Preop® (total: 400 ml). b. Antibiotic prophylaxis. Operating room
A. Surgeons: no drainage; nasogastric tube, if needed, to be removed before extubation; if possible use transverse or
curved incisions in open surgery. B Anaesthetists:
-Maintenance: Oxygen/air with FiO2 >80%. -Monitoring: routine. Only use arterial/central catheter if unavoidable. -Fluids: maintenance with Hartmann (5 cc/kg/h). Bolus of gelofusine (250 cc). Maintain Hb > 8.0 g/dl. - Optimise stroke volume via oesophageal Doppler:
-250 cc of fluid in bolus; if SV > 10% repeat until this figure is not reached. Provide no further bolus unless SV falls or
there is blood loss. -If hypotension remains after SV correction, use a vasoconstrictor. -Consider use of inotropic agents if peak velocity descends and clinical signs suggest ventricular function deficit. -Temperature: use liquid heater and heating blanket. Day of surgery (recovery
room)
-Mask with high oxygen flow for 2 h independent of saturation. Follow with nasal cannulae to maintain SpO2 > 95%. -Maintain mean blood pressure >65 mmHg. Enhanced recovery program protocol If blood pressure low provide 250 cc gelofusine and reassess. Day of surgery (ward)
-In the evening sit patient in seat for at least 2 h. -Liquid diet (800-1000 ml). Include two bricks of high protein/high calorie hospital dietary preparation (specific for
postoperative period). -Minimum diuresis (500 cc in first 24 h). -Analgesia: 1 g paracetamol/6 h. Postoperative day 1
-Liquid diet, at least 2 l, including 3 bricks of high protein/high calorie hospital dietary preparation. -Mobilisation; patient seated at least 6 h per day. -Suspend IV fluid if tolerated. Heparin injection for maintaining patency of intermittent infusion devices. -Maintain epidural analgesia pump if one in place. -Paracetamol 1 g/6 h. -Lactulose 1 sachet/12 h (preferably magnesium-based). -Assess meeting of discharge criteria: only oral analgesia, mobilisation reaching presurgical level, toleration of solid food,
gases passed, stools passed, no nausea, and patient agrees to discharge. Postoperative day 2
-Suspend epidural catheter; begin with NSAIDS; diet soft/normal; mobilisation on demand; remove urinary catheter and
assess meeting of discharge criteria. Postoperative day 3
-Check general status; assess meeting of discharge criteria and take decision in this respect. Follow up
-Telephone monitoring for 48 h. -First out-patient visit 10-14 days after surgery. SV: stroke volume; IV: intravenous; Hb: haemoglobin. SV: stroke volume; IV: intravenous; Hb: haemoglobin. postoperative vomiting and nausea, and the use of epi-
dural anaesthesia. summarises the protocol characteristics of the ERAS
followed. Although not explicitly required by the ERAS, the fol-
lowing variables were also recorded: opioid-free pain
control, the use of prophylactic medication for Patients were discharged following the criteria estab-
lished by the ERAS protocol. All patients were followed-
up for at least three months. Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Page 4 of 8 Page 4 of 8 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 An online database http://www.ftsurgery.com/ was pre-
pared for the collection of data from the different centres. The median age of the patients was 69 years (35-88);
51.5% were women (152 patients). The patients were
distributed by ASA class as follows: I 10.2% (30), II
50.2% (148) and III 39.7% (117). Patients, surgery and postoperative treatment Patients, surgery and postoperative treatment
Of all patients who initially met criteria for inclusion, 16
were excluded, 9 patients were in intensive care at the
discretion of the anaesthetist and 7 received an
unscheduled ileostomy. A total of 300 patients were
finally included. Main features of the protocol and their compliance rates
Table 3 shows the compliance rates for the main fea-
tures outlined in the ERAS protocol, as well as for the
other features not explicitly included in the protocol. Table 2 shows the characteristic of the patients
included, the surgical techniques used and the surgical
procedures followed. Table 3 Compliance with perioperative treatment and
postoperative recovery after colon resection within the
context of the enhanced recovery program followed
Perioperative treatment variable mentioned in ERAS protocol
Number Percentage
Preoperative
Perioperative information
290
99
No colon preparation
246
82.8
Carbohydrate-rich drinks on day before surgery1 187
65.2
Carbohydrate -rich drinks before surgery2
182
63
Surgery
Goal directed fluids (Cardio-Q)3
138
46.3
No hypothermia
242
84.3
No drainage
146
53.3
Postoperative
No nasogastric tube
222
77.1
Early mobilisation
128
44.6
Early taking of fluids by mouth4
116
40.6
Variable not explicitly mentioned in ERAS protocol
Number Percentage
Opioid-free pain control5
206
78.3
Prophylactic medication for nausea and
vomiting
76
26.8
Epidural anaesthesia
106
38.8
1: 40.3% received two, 59.7% received three or four; 2: 14.8% received one,
85.2% received two; 3: Mean fluid volume received was 1742.5 ml; 4: 46.8%
received one, 50% received two, and 3.2% received more than two; 5: 42.8%
of patients used patient controlled analgesia. Table 3 Compliance with perioperative treatment and
postoperative recovery after colon resection within the
context of the enhanced recovery program followed Table 3 Compliance with perioperative treatment and
postoperative recovery after colon resection within the
context of the enhanced recovery program followed
Perioperative treatment variable mentioned in ERAS protocol
Number Percentage
Preoperative
Perioperative information
290
99
No colon preparation
246
82.8
Carbohydrate-rich drinks on day before surgery1 187
65.2
Carbohydrate -rich drinks before surgery2
182
63
Surgery
Goal directed fluids (Cardio-Q)3
138
46.3
No hypothermia
242
84.3
No drainage
146
53.3
Postoperative
No nasogastric tube
222
77.1
Early mobilisation
128
44.6
Early taking of fluids by mouth4
116
40.6 Data analysis Some 63.6% (171) of the patients underwent laparo-
scopic surgery and 36.4% (98) open surgery. Some
15.3% of the laparoscopically intervened patients
required conversion to laparotomy (22). Dichotomous variables were recorded as absolute fre-
quencies (number of cases) and relative frequencies
(percentages). It should be noted that this work reports
frequencies and percentages with respect to the data
made available; therefore they do not always refer to all
patients. Continuous variables were recorded as means
and standard deviations (SD) or median plus maximum
and minimum values, depending on whether or not
their distribution was normal (determined by the Kol-
mogorov-Smirnov test). All analyses were made using
18 version SPSS software. Sigmoidectomy and right hemicolectomy made up the
majority of procedures performed (62.5%). Some 5.2% of
patients (15) were subjected to preoperative radiother-
apy. Laparotomy was medial in 88.8% and transversal in
11.2% of patients so intervened. The mean preoperative hospital stay was 0.82 days
(SD: 1.93). The mean duration of surgery was 155.4
min. The median length of time spent in the recovery
room was 240 min (60-1500). Results Patients, surgery and postoperative treatment Table 2 Patient characteristics and surgical techniques
and procedures followed Table 2 Patient characteristics and surgical techniques
and procedures followed Table 2 Patient characteristics and surgical techniques
and procedures followed
Patients characteristics (n = 300)
Values
Age (years)
69
35-88
Sex
Female
152
51.5
Male
143
48.5
Preoperative stay (days)
0.82
1.93
Surgical risk: ASA
I
30
10.2
II
148
50.2
III
117
39.7
Surgical technique
Laparoscopy
171
63.6
Conventional
98
36.4
Surgical procedure
Right hemicolectomy
98
35.6
Sigmoidectomy
74
26.9
Anterior resection
67
24.4
Left hemicolectomy
27
9.8
Transverse resection
5
1.8
Subtotal colectomy
4
1.5
Q
i
i
i bl
d
di
l
i i
d
i Variable not explicitly mentioned in ERAS protocol
Number Percentage
Opioid-free pain control5
206
78.3
Prophylactic medication for nausea and
vomiting
76
26.8
Epidural anaesthesia
106
38.8
1: 40.3% received two, 59.7% received three or four; 2: 14.8% received one,
85.2% received two; 3: Mean fluid volume received was 1742.5 ml; 4: 46.8%
received one, 50% received two, and 3.2% received more than two; 5: 42.8%
of patients used patient controlled analgesia. Variable not explicitly mentioned in ERAS protocol
Number Percentage
Opioid-free pain control5
206
78.3
Prophylactic medication for nausea and
vomiting
76
26.8
Epidural anaesthesia
106
38.8 Variable not explicitly mentioned in ERAS protocol Quantitative variables are expressed as medians plus minimum and maximum
values; qualitative variables are expressed as absolute numbers and
percentages. Preoperative stay is expressed as mean and standard deviation. Page 5 of 8 Page 5 of 8 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Ramírez et al. Table 2 Patient characteristics and surgical techniques
and procedures followed BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Table 4 Local and general morbidity following colon
resection within the enhanced recovery program
followed
Total (n = 300)
Number
Percentage
Surgical complications
71
23.7
Wound infection
36
12
Paralytic ileus
30
10
Anastomosis leakage
13
4.3
Perioperative haemorrhage
5
1.7
Evisceration
3
1
Perforation
0
0
Other surgical complications
10
3.3
Septic complications
33
11
Abdominal abscess
13
4.3
Urine infection
11
3.7
Respiratory infection
11
3.7
Peritonitis
7
2.3
Sepsis
3
1
Catheter sepsis
2
0.7
Necrotising fascitis
0
0
Other septic complications
3
1
Medical complications
29
9.7
Respiratory distress
8
2.7
Respiratory failure
8
2.7
Cardiac complication
5
1.7
Pulmonary oedema
3
1
Acute urine retention
3
1
Pulmonary embolism
1
0.3
Myocardial infarction
0
0
Cerebrovascular accident
0
0
Venous thrombosis
0
0
Other medical complications
10
3.3
All complications
89
29.7
Mortality
3
1
Note: some patients presented more than one symptom. Table 4 Local and general morbidity following colon
resection within the enhanced recovery program
followed Table 4 Local and general morbidity following colon
resection within the enhanced recovery program
followed The overall compliance to protocol was approximately
65%, but varied widely in its different components. Postoperative morbidity/mortality Table 4 summarises the complications encountered. The
most common were surgical (23.7% of patients; 71), fol-
lowed by septic 11% (33) and other medical complica-
tions
9.7%
(29). The
most
common
surgical
complications were wound infection (12%; 36), paralytic
ileus (10%; 30; median paralysis time 3 days [2-8]), ana-
stomosis leakage (4.3%; 13) and perioperative haemor-
rhage
(1.7%;
5). The
most
common
medical Figure 1 Length of postoperative stay of patients who
underwent colon surgery within the context of the present
enhanced recovery program. Note: some patients presented more than one symptom. complications were respiratory distress (2.7%; 8),
respiratory failure (2.7%%; 8) and cardiac arrhythmia or
ischemia (1.7%; 5). Finally, the most common septic
complications were abdominal abscess (4.3%; 13), urine
infection (3.7%; 11), respiratory infection (3.7%; 11) and
peritonitis (2.3%; 7). Three patients (1%), all of whom required repeat sur-
gery, died. Two of these died during their hospital stay
due to multi-organ failure and sepsis, and one from can-
cer following discharge. Figure 1 Length of postoperative stay of patients who
underwent colon surgery within the context of the present
enhanced recovery program. Postoperative hospital stay and readmission rate The median length of postoperative hospital stay was 6
days (3-89 days). Figure 1 shows the distribution of
length of postoperative stay. The mean postoperative
length of stay according to surgical procedure was: right
hemicolectomy and sigmoidectomy 8 days, left hemico-
lectomy 6 days, and transverse resection and subtotal
colectomy 5 days. Some 2.7% of the patients (8) were readmitted follow-
ing discharge for medical or surgical reasons. Three
patients re-presented with febrile syndrome, two with
nosocomial pneumonia, and three with pulmonary
thromboembolism, diarrhoea with hyponatremia or
abdominal wall abscess. Some 7% of the patients (21) required repeat surgery. The causes included dehiscence (12 patients), fistula or
anastomatic leakage (three patients), evisceration (two
patients), and abdominal pain, haemoperitoneum, ische-
mia and intestinal occlusion (one patient each). The
median post-repeat-operative length of hospital stay was
6 days (1-20). Discussion
h
f h Each of the steps outlined in the present ERAS is based
on scientific evidence. For example, patient education is
reported to be important in the response to surgery. Now-classic studies [3] have shown that informed
patients require less analgesia in the postoperative per-
iod and indeed experience significantly less pain than
uninformed patients. More recent work has shown that
adequate preoperative information reduces patient anxi-
ety before surgery and may also hasten postsurgical
recovery [3,13,14]. The items of the protocol with less compliance were
early oral fluid administration, goal-directed fluid ther-
apy and early mobilization. The reason why these items
obtained different compliance with the protocol could
be the taste of oral fluid, rejection by patients, unavail-
ability of devices and temporary employment of some
healthcare providers involved in the ERAS. Probably the
implementation could improve involving and training all
the professionals who assist the patients included in the
protocol and identifying these patients with signboards
on bedside. y
A number of studies on programmed colon surgery
have brought into doubt the need for preoperative
mechanical cleansing of the intestine [3,15]. The need
for strict preoperative fasting has also been recently
questioned. Most clinical practice guides suggest a per-
iod of absolute fasting of between two and six hours,
but recent studies indicate that taking a carbohydrate-
rich drink before surgery may reduce the endocrine
catabolic response and improve insulin resistance [3,16],
improving surgical results and hastening recovery. The
present ERAS included the administration of carbohy-
drate-rich drinks (4 × 200 ml) one day prior to surgery
plus two further such drinks (2 × 200 ml) on the morn-
ing of surgery. The protocol also included the adminis-
tration of goal directed fluids made possible by the
standard use of oesophageal Doppler monitoring
[3,17-21], temperature control to avoid hypothermia [3],
and the non-routine use of a nasogastric tube; meta-
analyses of several trials suggest the latter may reduce
pulmonary complications [3,22]. A further measure was Missing values were most important for the variable
surgical approach with 6.2% of them, being lower in the
other variables. The rates of complications and mortality recorded were
similar to those reported by other authors in randomized
controlled trials [5,8-11]. In the present work the most
common complication was wound infection. These pro-
grammes do not, therefore, appear to place the patient at
any extra risk. Symptoms delaying discharge Table 5 records the symptoms that delayed discharge. Some 31.3% (94) of all patients presented some such Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Page 6 of 8 Page 6 of 8 Table 5 Symptoms delaying discharge
Total (n = 300)
Number of patients
Percentage
Vomiting or nausea
36
12
Fatigue
20
6.7
Fever of unknown origin
14
4.7
Constipation
9
3
Dizziness
6
2
Pain
6
2
Urine retention
5
1.7
Depression, confusion
4
1.3
Diarrhoea
3
1
Scant diuresis
3
1
Other symptoms
26
8.7
Total
94
31.3
Note: some patients presented more than one symptom. Table 5 Symptoms delaying discharge
Total (n = 300)
Number of patients
Percentage
Vomiting or nausea
36
12
Fatigue
20
6.7
Fever of unknown origin
14
4.7
Constipation
9
3
Dizziness
6
2
Pain
6
2
Urine retention
5
1.7
Depression, confusion
4
1.3
Diarrhoea
3
1
Scant diuresis
3
1
Other symptoms
26
8.7
Total
94
31.3
Note: some patients presented more than one symptom. Table 5 Symptoms delaying discharge the avoidance of routinely using drainage; several rando-
mised trials have suggested that drainage is of no benefit
[3,23,24]. Drainage can be avoided in most patients or
limited to a short period, facilitating early mobilisation
[3], a measure also called for by the ERAS followed. Finally, although taking food orally is commonly limited
in the postoperative period, a number of studies have
shown that it is safe even after colon surgery involving
anastomosis [3,25,26]; it was therefore included in the
present ERAS. Additional variables not explicitly included in the
ERAS were also measured: use of prophylactic medica-
tion for postoperative vomiting and nausea, use of epi-
dural anaesthesia [3], and opioid-free pain control, it has
been reported that opioid-free or opioid-reduced analge-
sia may hasten recovery [3]. Note: some patients presented more than one symptom. When protocols such as the present are implanted, the
goal is that there should be full compliance with all
measures outlined. However, full compliance is com-
monly very difficult to achieve [12]. In the present work
the overall compliance rate was 65%, but varied widely
in its different components. Patients received informa-
tion in nearly all cases, while compliance with the provi-
sion of early postoperative fluids seemed particularly
difficult. symptom, 22% (66) suffered one, 6.3% (19) suffered two,
2.7% (8) suffered three, and 0.3% (1) suffered four. Symptoms delaying discharge The
most common symptoms were vomiting (12%; 36), dys-
pnoea (6.7%; 20) and fever of unknown origin (4.7%; 14). Acknowledgements
Th
h
h
k The authors thank specially to all members of the Spanish working group
on fast track surgery: Emilio Maseda, Rafael Uña, Damián García-Olmo
(Hospital Universitario La Paz, Madrid, Spain); Carlos Moreno, Marina
Manzanera, Francisco A Quezada, Telesfora Sempere (Hospital General La
Mancha Centro, Alcázar de San Juan, Spain); Eva Llacer, Carlos Maristany,
Arantxa Muñoz, Albert Navarro (Hospital Universitario Mútua Terrassa,
Terrassa, Spain); Javier Cerdán, Gonzalo Sanz, Pedro Moral, Fernando Esteban
(Hospital Clínico San Carlos, Madrid, Spain); Pablo Royo, Vicente Aguilella,
Julia Guillen, Tomás Ruiz, José Cuartero, Mariano Martínez, José M Ramírez
(Hospital Clínico Universitario Lozano Blesa, Zaragoza, Spain); Enrique
Moncada, Manuel Núñez, Luis C Luna, José E Casal (Hospital Do Meixoeiro,
Vigo, Spain); Roger Cabezali, Carlos Emparan, Pablo Soriano, Javier Isla
(Fundación Hospital Calahorra, Calahorra, Spain); Antonio Arroyo, Alessandro
Garcea, Pilar Serrano, José L Muñoz, Elena Miranda (Hospital General
Universitario de Elche, Elche, Spain); José V. Roig, Francisco Villalba, Antonio
Salvador, Alfonso Garcia-Fabrique (Hospital General Universitario de Valencia,
Valencia, Spain); Luis M Jiménez, Elena Monge, Irene Hidalgo, Emilio Del
Valle (Hospital General Universitario Gregorio Marañón, Madrid, Spain); Rafael
Morales, José Noguera (Hospital Son Llatzer, Palma de Mallorca, Spain);
Xavier Viñas, Enric Macarulla, Victor Murga, Ana Pedregosa (Hospital
d’Igualada, Igualada, Spain); Juan A Blasco, Sergio Maeso-Martínez and Daniel
Callejo (Health Technology Assessment, Agencia Laín Entralgo, Madrid,
Spain). The authors thank Adrian Burton for help with the English language
manuscript. 10. Gatt M, Anderson AD, Reddy BS, Hayward-Sampson P, Tring IC, MacFie J:
Randomized clinical trial of multimodal optimization of surgical care in
patients undergoing major colonic resection. Br J Surg 2005,
92:1354-1362. 10. Gatt M, Anderson AD, Reddy BS, Hayward-Sampson P, Tring IC, MacFie J:
Randomized clinical trial of multimodal optimization of surgical care in
patients undergoing major colonic resection. Br J Surg 2005,
92:1354-1362. 11. Khoo CK, Vickery CJ, Forsyth N, Vinall NS, Eyre-Brook IA: A prospective
randomized controlled trial of multimodal perioperative management
protocol in patients undergoing elective colorectal resection for cancer. Ann Surg 2007, 245:867-872. 12. Maessen J, Dejong CH, Hausel J, Nygren J, Lassen K, Andersen J, Kessels AG,
Revhaug A, Kehlet H, Ljungqvist O, et al: A protocol is not enough to
implement an enhanced recovery programme for colorectal resection. Br
J Surg 2007, 94:224-231. 13. Daltroy LH, Morlino CI, Eaton HM, Poss R, Liang MH: Preoperative
education for total hip and knee replacement patients. Arthritis Care Res
1998, 11:469-478. 14. Discussion
h
f h Our results are similar to previous multi-
center studies [27,28] in terms of surgical complications,
mortality and readmission rate. The surgical complica-
tion rate was 24% compared to the 14.1% reported by
Schwenk et al [27] and 20% reported by Braumann et al
[28]. The mortality rate was 1% compared to the 0.8%
reported by Schwenk et al [27] and 0,4% reported by
Braumann et al [28]. The readmission rate was 2.7%
compared to the 3.9% reported by Schwenk et al [27] and
4% reported by Braumann et al [28]. Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Page 7 of 8 Page 7 of 8 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Finally, the use of the present ERAS was associated
with a preoperative hospital stay of fewer than 24 h and
an overall mean stay of 6 days. In other recently pub-
lished Spanish multicenter study, including data from 50
hospitals, the mean postoperative stay after colorectal
resection was 12.36 days [29]. p
g
The authors declare that they have no competing interests. Received: 7 October 2010 Accepted: 14 April 2011
Published: 14 April 2011 Received: 7 October 2010 Accepted: 14 April 2011
Published: 14 April 2011 Received: 7 October 2010 Accepted: 14 April 2011
Published: 14 April 2011 g
y
22. Cheatham ML, Chapman WC, Key SP, Sawyers JL: A meta-analysis of
selective versus routine nasogastric decompression after elective
laparotomy. Ann Surg 1995, 221:469-476. 22. Cheatham ML, Chapman WC, Key SP, Sawyers JL: A meta-analysis of
selective versus routine nasogastric decompression after elective
laparotomy. Ann Surg 1995, 221:469-476. Acknowledgements
Th
h
h
k Klafta JM, Roizen MF: Current understanding of patients’ attitudes toward
and preparation for anesthesia: a review. Anesth Analg 1996,
83:1314-1321. 15. Burke P, Mealy K, Gillen P, Joyce W, Traynor O, Hyland J: Requirement for
bowel preparation in colorectal surgery. Br J Surg 1994, 81:907-910. 16. Nygren J, Soop M, Thorell A, Sree NK, Ljungqvist O: Preoperative oral
carbohydrates and postoperative insulin resistance. Clin Nutr 1999,
18:117-120. Authors’ contributions Authors contributions
Design: JMR, JVR, JEC, FE, JAB, DC and SM. Data collection: JMR, JVR, JEC and FE. Data analysing: SM, DC, JAB and JMR. Writing of manuscript: SM, JAB, JMR and DC. All authors have read and approve the final manuscript. 19. Mowatt G, Houston G, Hernandez R, de VR, Fraser C, Cuthbertson B, Vale L:
Systematic review of the clinical effectiveness and cost-effectiveness of
oesophageal Doppler monitoring in critically ill and high-risk surgical
patients. Health Technol Assess 2009, 13:1-118. Design: JMR, JVR, JEC, FE, JAB, DC and SM. Design: JMR, JVR, JEC, FE, JAB, DC and SM. Data collection: JMR, JVR, JEC and FE. g
Data collection: JMR, JVR, JEC and FE. Data analysing: SM, DC, JAB and JMR. Writing of manuscript: SM, JAB, JMR and DC. 20. Noblett SE, Snowden CP, Shenton BK, Horgan AF: Randomized clinical trial
assessing the effect of Doppler-optimized fluid management on
outcome after elective colorectal resection. Br J Surg 2006, 93:1069-1076. All authors have read and approve the final manuscript. All authors have read and approve the final manuscript. Conclusion The present ERAS posed no risk to patients in terms of
morbidity, mortality and shortened their hospital stay. The present results show that these programmes can be
of benefit to patients and, by reducing hospital costs,
may benefit society as a whole. p
g
g
8. Stephen AE, Berger DL: Shortened length of stay and hospital cost
reduction with implementation of an accelerated clinical care pathway
after elective colon resection. Surgery 2003, 133:277-282. 9. Delaney CP, Zutshi M, Senagore AJ, Remzi FH, Hammel J, Fazio VW:
Prospective, randomized, controlled trial between a pathway of
controlled rehabilitation with early ambulation and diet and traditional
postoperative care after laparotomy and intestinal resection. Dis Colon
Rectum 2003, 46:851-859. Author details
1
f 17. Conway DH, Mayall R, bdul-Latif MS, Gilligan S, Tackaberry C: Randomised
controlled trial investigating the influence of intravenous fluid titration
using oesophageal Doppler monitoring during bowel surgery. Anaesthesia 2002, 57:845-849. 1Department of Colorectal Surgery, Hospital Clínico Universitario Lozano
Blesa, Zaragoza, Spain. 2Health Technology Assessment, Agencia Laín
Entralgo, Madrid, Spain. 3Department of Surgery, Hospital General
Universitario de Valencia, Valencia, Spain. 4Department of Surgery, Hospital
Do Meixoeiro, Vigo, Spain. 5Department of Surgery, Hospital Clínico San
Carlos, Madrid, Spain. 18. Gan TJ, Soppitt A, Maroof M, el-Moalem H, Robertson KM, Moretti E,
Dwane P, Glass PS: Goal-directed intraoperative fluid administration
reduces length of hospital stay after major surgery. Anesthesiology 2002,
97:820-826. 18. Gan TJ, Soppitt A, Maroof M, el-Moalem H, Robertson KM, Moretti E,
Dwane P, Glass PS: Goal-directed intraoperative fluid administration
reduces length of hospital stay after major surgery. Anesthesiology 2002,
97:820-826. 1.
Schoetz DJ Jr, Bockler M, Rosenblatt MS, Malhotra S, Roberts PL, Murray JJ,
Coller JA, Rusin LC: “Ideal” length of stay after colectomy: whose ideal?
Dis Colon Rectum 1997, 40:806-810. Competing interests 21. Wakeling HG, McFall MR, Jenkins CS, Woods WG, Miles WF, Barclay GR,
Fleming SC: Intraoperative oesophageal Doppler guided fluid
management shortens postoperative hospital stay after major bowel
surgery. Br J Anaesth 2005, 95:634-642. References 23. Merad F, Yahchouchi E, Hay JM, Fingerhut A, Laborde Y, Langlois-Zantain O:
Prophylactic abdominal drainage after elective colonic resection and
suprapromontory anastomosis: a multicenter study controlled by 23. Merad F, Yahchouchi E, Hay JM, Fingerhut A, Laborde Y, Langlois-Zantain O:
Prophylactic abdominal drainage after elective colonic resection and
suprapromontory anastomosis: a multicenter study controlled by Page 8 of 8 Page 8 of 8 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 randomization. French Associations for Surgical Research. Arch Surg 1998,
133:309-314. randomization. French Associations for Surgical Research. Arch Surg 1998,
133:309-314. 24. Urbach DR, Kennedy ED, Cohen MM: Colon and rectal anastomoses do
not require routine drainage: a systematic review and meta-analysis. Ann
Surg 1999, 229:174-180. 25. Carr CS, Ling KD, Boulos P, Singer M: Randomised trial of safety and
efficacy of immediate postoperative enteral feeding in patients
undergoing gastrointestinal resection. BMJ 1996, 312:869-871. 26. Di Fronzo LA, Cymerman J, O’Connell TX: Factors affecting early
postoperative feeding following elective open colon resection. Arch Surg
1999, 134:941-945. 27. Schwenk W, Gunther N, Wendling P, Schmid M, Probst W, Kipfmuller K,
Rumstadt B, Walz MK, Engemann R, Junghans T: “Fast-track” rehabilitation
for elective colonic surgery in Germany–prospective observational data
from a multi-centre quality assurance programme. Int J Colorectal Dis
2008, 23:93-99. 28. Braumann C, Guenther N, Wendling P, Engemann R, Germer CT, Probst W,
Mayer HP, Rehnisch B, Schmid M, Nagel K, Schwenk W, Fast-Track Colon II
Quality Assurance Group: Multimodal perioperative rehabilitation in
elective conventional resection of colonic cancer: results from the
German Multicenter Quality Assurance Program ‘Fast-Track Colon II’. Dig
Surg 2009, 26:123-9. 29. Rodríguez-Cuellar E, Ruiz López P, Romero Simó M, Landa García JI, Roig
Vila JV, Ortiz Hurtado H: Analysis of the quality of surgical treatment of
colorectal cancer, in 2008. A national study. Cir Esp 2010, 88:238-46. Ramírez et al. BMC Surgery 2011, 11:9
http://www.biomedcentral.com/1471-2482/11/9 Pre-publication history The pre-publication history for this paper can be accessed here:
http://www.biomedcentral.com/1471-2482/11/9/prepub The pre-publication history for this paper can be accessed here:
http://www.biomedcentral.com/1471-2482/11/9/prepub doi:10.1186/1471-2482-11-9
Cite this article as: Ramírez et al.: Enhanced recovery in colorectal
surgery: a multicentre study. BMC Surgery 2011 11:9. doi:10.1186/1471-2482-11-9
Cite this article as: Ramírez et al.: Enhanced recovery in colorectal
surgery: a multicentre study. BMC Surgery 2011 11:9. doi:10.1186/1471-2482-11-9
Cite this article as: Ramírez et al.: Enhanced recovery in colorectal
surgery: a multicentre study. BMC Surgery 2011 11:9. Submit your next manuscript to BioMed Central
and take full advantage of:
• Convenient online submission
• Thorough peer review
• No space constraints or color figure charges
• Immediate publication on acceptance
• Inclusion in PubMed, CAS, Scopus and Google Scholar
• Research which is freely available for redistribution
Submit your manuscript at
www.biomedcentral.com/submit
|
https://openalex.org/W2922241779
|
http://e-journal.lp2m.uinjambi.ac.id/ojp/index.php/iltizam/article/download/90/41
|
Indonesian
| null |
PENGARUH INVESTASI DAN PENGELUARAN PEMERINTAH TERHADAP PERTUMBUHAN EKONOMI PROVINSI JAMBI (STUDI TAHUN 2012-2015)
|
Iltizam Journal of Shariah Economic Research
| 2,017
|
cc-by-sa
| 6,286
|
1 | Iltizam Journal Of Shariah Economic Research, Vol. 1, No. 1, 2017 1 | Iltizam Journal Of Shariah Economic Research, Vol. 1, No. 1, 2017 Abstract Studi ini bertujuan untuk menentukan pengaruh investasi dan pengeluaran
pemerintah terhadap pertumbuhan ekonomi Provinsi Jambi dari tahun
2012 sampai tahun 2015. Metode penelitian yang digunakan adalah
pendekatan kuantitatif. Sebagai tujuan penelitian diantaranya adalah untuk
mengetahui pengaruh investasi terhadap pertumbuhan ekonomi, pengaruh
pengeluaran pemerintah terhadap pertumbuhan ekonomi serta pengaruh
investasi dan pengeluaran pemerintah terhadap pertumbuhan ekonomi
Provinsi Jambi. Berdasarkan penelitian yang dilakukan diperoleh hasil dan
kesimpulan sebagai berikut: pertama, bahwa investasi berpengaruh
signifikan terhadap pertumbuhan ekonomi Provinsi Jambi, kedua
pengeluaran pemerintah berpengaruh signifikan terhadap pertumbuhan
ekonomi Provinsi Jambi, ketiga secara simultan investasi dan pengeluaran
pemerintah berpengaruh signifikan terhadap pertumbuhan ekonomi
Provinsi Jambi, dengan nilai R Square yang diperoleh 0,492, artinya variabel
investasi dan pengeluaran pemerintah mempengaruhi pertumbuhan
ekonomi Provinsi Jambi sebesar 49,2 % sedangkan sisanya 50,8 %
pertumbuhan ekonomi Provinsi Jambi dipengaruhi oleh faktor lain. PENGARUH INVESTASI DAN PENGELUARAN PEMERINTAH
TERHADAP PERTUMBUHAN EKONOMI PROVINSI JAMBI
(STUDI TAHUN 2012-2015) Ambok Pangiuk
Fakultas Ekonomi dan Bisnis Islam UIN STS Jambi
ambokpangiuk1975@gmail.com Kata Kunci : Pertumbuhan Ekonomi, Investasi dan Pengeluaran Pemerintah 3 Rozalinda, Ekonomi Islam Teori dan Aplikasinya pada Aktivitas Ekonomi, (Jakarta: Raja Grafindo Persada,
2014), hlm. 139 1 Mohammad Rizal Mubaroq dkk, “Pengaruh Investasi Pemerintah, Tenaga Kerja, dan Desentralisasi Fiskal
Terhadap Pertumbuhan Ekonomi Kabupaten Di Indonesia Tahun 2007 – 2010”, Jurnal M. Rizal M Met UNPAD, (Januari
2013), hlm. 1 )
2 BPS Provinsi Jambi, Analisis Pertumbuhan Ekonomi Provinsi Jambi 2014, (Jambi, BPS Provinsi Jambi, 2014),
hlm. 6 ),
4 Moh.Arsjad Anwar dkk, Ekonomi Indonesia, Masalah dan prosfek 1989/1990, (Jakarta: Universitas
Indonesia Press, 1989), hlm. 47 q
,
g
,
g
j ,
Pertumbuhan Ekonomi Kabupaten Di Indonesia Tahun 2007 – 2010”, Jurnal M. Rizal M Met UNPAD, (Januari
m. 1 3 Rozalinda, Ekonomi Islam Teori dan Aplikasinya pada Aktivitas Ekonomi, (Jakarta: Raja Grafindo Persada,
2014), hlm. 139
4 Moh.Arsjad Anwar dkk, Ekonomi Indonesia, Masalah dan prosfek 1989/1990, (Jakarta: Universitas
Indonesia Press 1989) hlm 47 1 Mohammad Rizal Mubaroq dkk, “Pengaruh Investasi Pemerintah, Tenaga Kerja, dan Desentralisasi Fiskal PENDAHULUAN Pertumbuhan ekonomi merupakan ukuran
atas perkembangan atau
kemajuan perekonomian dari suatu negara atau wilayah karena berkaitan erat
dengan kegiatan ekonomi masyarakat khususnya dalam hal peningkatan produksi Iltizam Journal Of Shariah Economic Research, Vol. 1, No. 1, 2017
Fakultas Ekonomi dan Bisnis Islam UIN Sulthan Thaha Saifuddin Jambi
E-ISSN: 2598-2540, P-ISSN: 2598-2222
Availible Online http://e-journal.lp2m.uinjambi.ac.id/ojp/index.php/iltizam Iltizam Journal Of Shariah Economic Research, Vol. 1, No. 1, 2017 J
Fakultas Ekonomi dan Bisnis Islam UIN Sulthan Thaha Saifuddin Jambi
E-ISSN: 2598-2540, P-ISSN: 2598-2222 J
Fakultas Ekonomi dan Bisnis Islam UIN Sulthan Thaha Saifuddin Jambi
E-ISSN: 2598-2540, P-ISSN: 2598-2222 ,
Availible Online http://e-journal.lp2m.uinjambi.ac.id/ojp/index.php/iltizam A m b o k P a n g i u k - P e n g a r u h I n v e s t a s i | 2 barang dan jasa. Peningkatan produksi tersebut kemudian diharapkan dapat
memberikan efek yang mampu meningkatkan kesejahteraan masyarakat.1 Pertumbuhan ekonomi dalam sistem pemerintahan daerah biasanya di
indikasikan dengan meningkatnya produksi barang dan jasa yang diukur melalui
Produk Domestik Regional Bruto (PDRB).PDRB didefinisikan sebagai nilai
tambah bruto seluruh barang dan jasa yang tercipta atau dihasilkan diwilayah
domestik suatu negara yang timbul akibat berbagai aktivitas ekonomi dalam suatu
periode tertentu.2 Kegiatan ekonomi dalam suatu negara atau wilayah akan optimal
jika terdapat aktifitas pemerintah didalamnya, karena pemerintah diberikan
kewenangan dalam mengatur pendapatan melalui penarikan pajak pendapatan
BUMN, selain itu pemerintah juga diberikan kewenangan untuk membelanjakan
anggaran untuk kepentingan masyarakat misalnya memberikan subsidi dan
melakukan pembangunan infrastuktur.3 Pengeluaran pemerintah merupakan indikator besarnya kegiatan pemerintah,
apabila semakin banyak kegiatan pemerintah maka semakin besar pula pengeluaran
pemerintah yang bersangkutan, namun ini bukanlah berarti bahwa pengeluaran
pemerintah harus selalu ditingkatkan tanpa memperhitungkan aspek efisiensinya. Pengeluaran pemerintah dapat mempengaruhi aktivitas ekonomi dikarenakan
pengeluaran pemerintah ini akan dapat menciptakan berbagai prasarana yang
dibutuhkan dalam proses pembangunan.4 Pengeluaran pemerintah yang tercermin
dalam Anggaran Pendapatan dan Belanja Daerah (APBD) bersumber dari bantuan
pusat dan Pendapatan Asli Daerah (PAD).APBD ini merupakan bentuk dari
akumulasi modal pemerintah yang digunakan untuk mendorong pertumbuhan
ekonomi suatu daerah. Adapun sasaran penggunaan pengeluaran pemerintah Pengeluaran pemerintah merupakan indikator besarnya kegiatan pemerintah,
apabila semakin banyak kegiatan pemerintah maka semakin besar pula pengeluaran
pemerintah yang bersangkutan, namun ini bukanlah berarti bahwa pengeluaran
pemerintah harus selalu ditingkatkan tanpa memperhitungkan aspek efisiensinya. 3 | Iltizam Journal Of Shariah Economic Research, Vol. 1, No. 5 Murdifin Haming & Salim Basalamah, Studi Kelayakan Investasi Proyek & Bisnis, (Jakarta: Bumi Aksara,
2010), hlm. 5
6 Sadono Sukirno, Ekonomi Pembangunan, (Jakarta: Kencana Prenadamedia Group, 2006), hlm. 257 5 Murdifin Haming & Salim Basalamah, Studi Kelayakan Investasi Proyek & Bisnis, (Jakarta: Bumi Aksara,
2010), hlm. 5 m. 5
6 Sadono Sukirno, Ekonomi Pembangunan, (Jakarta: Kencana Prenadamedia Group, 2006), hlm. 257 5 Murdifin Haming & Salim Basalamah, Studi Kelayakan Investasi Proyek & Bisnis, (Jakarta: Bumi Aksara,
5 2010), hlm. 5
6 Sadono Sukirno, Ekonomi Pembangunan, (Jakarta: Kencana Prenadamedia Group, 2006), hlm. 257 PENDAHULUAN 1, 2017 3 adalah untuk membiayai pembangunan di bidang sarana dan prasarana yang dapat
menunjang kelancaran usaha dan pemenuhan pelayanan masyarakat. Selain dari pada pengeluaran pemerintah Investasi yang memadai juga akan
berdampak positif terhadap kondisi perekonomian, karena dengan adanya investasi
yang memadai maka modal akan tersedia, semakin banyak investor yang
menanamkan modalnya tentunya akan meningkatkan produksi yang nantinya akan
menghasilkan output yang tinggi, serta akan menambah pendapatan daerah dari
pajak yang dibayarkan kepada pemerintah. Investasi merupakan keputusan
mengeluarkan dana pada saat sekarang ini untuk membeli aktiva riil ataupun aktiva
keuangan dengan tujuan untuk mendapatkan penghasilan yang lebih besar dimasa
yang akan datang.5 Pembentukan modal dipandang sebagai pengeluaran yang akan
menambah kesanggupan suatu perekonomian untuk menghasilkan barang, maupun
sebagai pengeluaran yang akan menambah permintaan efektif seluruh masyarakat.6 Adapun Indikator mengenai pengaruh investasi dan pengeluaran pemerintah
terhadap pertumbuhan ekonomi yang dibahas pada penelitian ini adalah sebagai
berikut : Indikator Pengaruh Investasi dan Pengeluaran Pemerintah terhadap
Pertumbuhan Ekonomi
Investasi
Pengeluaran
Pemerintah
Pertumbuhan Ekonomi
1. Sektor
Primer
:Perkebunan
Kelapa
Sawit
dan
Industri
Pengolahan,
Pertambangan,
Peternakan, Pengusahaan
Hutan Tanaman Industri
2. Sektor
Sekunder
:Industri Crumb Rubber,
Industri Kertas, Industri
Belanja Langsung:
- Belanja Pegawai
- Belanja Barang &
Jasa
- Belanja Modal
Indikator
pengukuran
pertumbuhan ekonomi :
- PDRB
atas
dasar
harga konstan
- PDRB
atas
dasar
harga berlaku
- Pendapatan Perkapita
Belanja
tidak Indikator Pengaruh Investasi dan Pengeluaran Pemerintah terhadap
Pertumbuhan Ekonomi A m b o k P a n g i u k - P e n g a r u h I n v e s t a s i | 4 Makanan, Industri Kimia
3. PENDAHULUAN Sektor
Tersier:Perdagangan
Jasa Penunjang
PertambanganListrik,
gas, dan air, Hotel dan
Restoran, Jasa Lainnya,
Perumahan, Jasa
Penyiaran Televisi
Langsung:
- Belanja Pegawai
- Belanja Subsidi
- Belanja Hibah
- Belanja
Bantuan
Sosial
- Belanja
bantuan
keuangan kepada
Prov/kab/kota
dan
pemerintah
desa
- Belanja
tidak
terduga
Sumber: BPS Provinsi Jambi & BPMD Untuk memberikan gambaran mengenai kondisi Investasi, Pengeluaran
Pemerintah dan pertumbuhan ekonomi Provinsi Jambi, maka dapat dilihat dalam
tabel berikut: Data Realisasi Investasi, Pengeluaran Pemerintah dan
Pertumbuhan Ekonomi Provinsi Jambi
No
Tahun
Investasi
Pengeluaran
Pemerintah
Pertumbuhan Ekonomi
1
2012
19.950.000,00
1.363.636,00
7,44
2
2013
25.000.000,00
1.740.363,00
7,88
3
2014
26.000.000,00
1.721.517,00
7,93
4
2015
30.000.000,00
1.741.503,00
4,95
Sumber: BPS Provinsi Jambi (data diolah) Data Realisasi Investasi, Pengeluaran Pemerintah dan
Pertumbuhan Ekonomi Provinsi Jambi Pada tabel tersebut terlihat bahwa realisasi investasi dari tahun 2012 hingga
tahun 2015 terus mengalami peningkatan, yang artinya investor memiliki
pandangan prospek yang baik terhadap keadaan investasi di Provinsi Jambi hingga
banyak investor yang tertarik untuk melakukan penanaman modal atau berinvestasi
di Provinsi Jambi. 5 | Iltizam Journal Of Shariah Economic Research, Vol. 1, No. 1, 2017 Pada tabel tersebut juga terlihat bahwa pengeluaran pemerintah pada tahun
2012 dan 2013 mengalami peningkatan, sedangkan ditahun 2014 pengeluaran
pemerintah sedikit mengalami penurunan dari tahun sebelumnya, dan selanjutnya
pengeluaran pemerintah kembali meningkat pada tahun 2015. Data Realisasi
Pengeluaran pemerintah yang kami sajikan disini adalah pengeluaran pemerintah
yang berupa belanja langsung, karena belanja langsung merupakan pengeluaran
yang lebih menekankan aspek efisiensi dan efektifitas dalam penggunaannya. Semakin tingginya pengeluaran yang dilakukan pemerintah menunjukkan bahwa
pemerintah terus berupaya untuk mensejahterakan rakyatnya dan untuk melakukan
pembangunan agar kondisi perekonomian Provinsi Jambi semakin meningkat. Sementara itu pada tabel juga terlihat bahwakondisi perekonomian Provinsi
Jambi dari tahun 2012 hingga tahun 2015 berfluktuasi, pertumbuhan ekonomi dari
tahun 2012 hingga tahun 2014 terus meningkat, dan peningkatan tertinggi adalah
pada tahun 2014 yaitu sebesar 7,93 % akan tetapi pertumbuhan ekonomi Provinsi
Jambi kembali melemah pada tahun 2015. Menurut Harrod dan Domar yang dikutip oleh Sayid syekh mengemukakan
bahwa investasi merupakan pengeluaran yang mempengaruhi kegiatan ekonomi,
maka investasi akan menambah jumlah barang modal didalam suatu masyarakat.7
Begitu pula Ibnu Khaldun mengatakan bahwasisi pengeluaran keuangan publik
merupakan hal yang sangat penting, karena pengeluaran pemerintah memiliki
peranan penting terhadap aktivitas ekonomi. Semakin banyak yang dibelanjakan
oleh pemerintah, semakin baik akibatnya bagi perekonomian.8 Pada umumnya para ekonom sepakat kalau tingkat investasi berkorelasi
positif dengan tingkat pertumbuhan ekonomi. 8 Adiwarman Azwar Karim, Sejarah Pemikiran Ekonomi Islam, (Jakarta: Raja Grafindo Persada, 2012), hlm. 8 Adiwarman Azwar Karim, Sejarah Pemikiran Ekonomi Islam, (Jakarta: Raja Grafindo Persada, 2012), hlm. 7 Sayid Syekh, Sekilas Pengantar Ilmu Ekonomi dan Pengantar Ekonomi Islam, (Jakarta: GP Press Group, 2013
100 7 Sayid Syekh, Sekilas Pengantar Ilmu Ekonomi dan Pengantar Ekonomi Islam, (Jakarta: GP Press Group, 2013),
hlm. 100 y
y
,
g
g
, (J
p,
),
hlm. 100
8 Adi
A
K i
S j
h P
iki
Ek
i I l
(J k
R j G fi d
P
d
2012) hl hlm. 100
8 Adiwarman Azwar Karim Sejarah Pemikiran Ekonomi Islam (Jakarta: Raja Grafindo Persada
2012) hlm 8 Adiwarman Azwar Karim, Sejarah Pemikiran Ekonomi Islam, (Jakarta: Raja Grafindo Persada, 20
410 Sayid Syekh, Sekilas Pengantar Ilmu Ekonomi dan Pengantar Ekonomi Islam, (Jakarta: GP Press Group, 2013),
hlm. 100
8 Adiwarman Azwar Karim, Sejarah Pemikiran Ekonomi Islam, (Jakarta: Raja Grafindo Persada, 2012), hlm. 7 Sayid Syekh, Sekilas Pengantar Ilmu Ekonomi dan Pengantar Ekonomi Islam, (Jakarta: GP Press Group, 2013),
hlm. 100
8 Adiwarman Azwar Karim, Sejarah Pemikiran Ekonomi Islam, (Jakarta: Raja Grafindo Persada, 2012), hlm.
410 PENDAHULUAN Secara sederhana, tingkat investasi
yang tinggi akan meningkatkan kapasitas produksi, yang pada akhirnya berujung
pada peningkatan pendapatan masyarakat. Teori juga mengatakan bahwa kenaikan 410 A m b o k P a n g i u k - P e n g a r u h I n v e s t a s i | 6 investasi dapat mendorong terjadinya pertumbuhan ekonomi.9 Akan tetapi fakta
yang terjadi di Provinsi Jambi menunjukkan bahwa pada saat investasi mengalami
pertumbuhan yang tinggi, tidak langsung dapat mendorong pertumbuhan ekonomi,
dan begitu pula sama halnya dengan pengeluaran pemerintah yang ketika
mengalami peningkatan ternyata tidak secara langsung meningkatkan pertumbuhan
ekonomi, begitu pula sebaliknya ketika investasi dan pengeluaran pemerintah
menurun tidak langsung mengakibatkan kondisi pertumbuhan ekonomi melemah. Hal ini terbukti dengan adanya data investasi dan pengeluaran pemerintah yang
mengalami peningkatan akan tetapi kondisi perekonomian Provinsi Jambi saat ini
mulai melemah atau mengalami penurunan dari tahun sebelumnya. p
,
10Kamus lengkap 10 Milyar, Djamaries, (Jakarta: Citra Harta Prima) hlm. 171 9 Efrizal Hasan, Syamsul Amar, Ali Anis, “Pengaruh Investasi, Angakatan kerja dan Pengeluaran Pemerintah
terhadap Pertumbuhan Ekonomi diProvinsi Sumatra Barat”, hlm. 1 g
p
y
j
(J
)
11 Ana Rokhmatussa’dyah & Suratman, Hukum Investasi & Pasar Modal, (Jakarta: Sinar Grafika, 2011), hlm p
,
10Kamus lengkap 10 Milyar, Djamaries, (Jakarta: Citra Harta Prima) hlm. 171 g
p
y ,
j
, (J
)
11 Ana Rokhmatussa’dyah & Suratman, Hukum Investasi & Pasar Modal, (Jak j
11 Ana Rokhmatussa’dyah & Suratman, Hukum Investasi & Pasar Modal, (Jakarta: Sinar Grafika, 2011), hlm. (J
j
)
13 Harry A. P. Sitaniapessy, “Pengaruh pengeluaran pemerintah terhadap PDRB dan PAD”, Jurnal Economia,
volume 9, nomor 1, (April 2013), hlm. 40 12 Salim dan Budi Sutrisno, Hukum Investasi di Indonesia, (Jakarta: Raja Grafindo Persada, 2008), hlm. 36-38 ( p
)
14 Badan Pusat Statistik Provinsi Jambi, Analisis Pertumbuhan Ekonomi Provinsi Jambi 2013, (Jambi: BP
nsi Jambi, 2013), hlm. 24 alim dan Budi Sutrisno, Hukum Investasi di Indonesia, (Jakarta: Raja Grafindo Persada, 2008), hlm. 36-38 1. Investasi Investasi berasal dari kata
Invest
yang berarti menanam atau
menginvestasikan uang atau modal.10 Dalam pasal 1 ayat (1) Undang-undang
nomor 25 tahun 2007 tentang penanaman modal disebutkan bahwa
penanaman modal diartikan sebagai segala bentuk kegiatan penanaman
modal, baik oleh penanam modal dalam negeri maupun penanam modal asing
untuk melakukan usaha diwilayah negara Republik Indonesia. Secara umum investasi atau penanaman modal dapat diartikan sebagai
suatu kegiatan yang dilakukan baik oleh pihak pribadi (Natural Person) maupun
badan hukum (Juridical Person) dalam upaya untuk meningkatkan dan/ atau
mempertahankan nilai modalnya, baik yang berbentuk uang tunai (Cash
Money), peralatan, aset tidak bergerak, hak atas kekayaan intelektual maupun
keahlian.11 3 7 | Iltizam Journal Of Shariah Economic Research, Vol. 1, No. 1, 2017 Investasi berdasarkan pembiayaannya merupakan investasi yang
didasarkan pada asal-usul investasi itu diperoleh. Investasi ini dibagi menjadi
dua macam, yaitu : Investasi berdasarkan pembiayaannya merupakan investasi yang
didasarkan pada asal-usul investasi itu diperoleh. Investasi ini dibagi menjadi
dua macam, yaitu : 1) Investasi yang bersumber dari modal asing (PMA), merupakan investasi
yang bersumber dari pembiayaan luar negeri. 2) Investasi yang bersumber dari modal dalam negeri (PMDN), merupakan
investasi yang bersumber dari pembiayaan dalam negeri.12 2) Investasi yang bersumber dari modal dalam negeri (PMDN), merupakan
investasi yang bersumber dari pembiayaan dalam negeri.12 2. Pengeluaran Pemerintah Pengeluaran pemerintah (goverment expenditure) adalah bagian dari
kebijakan fiskal yaitu suatu tindakan pemerintah untuk mengatur jalannya
perekonomian
dengan
cara
menentukan
besarnya
penerimaan
dan
pengeluaran pemerintah setiap tahunnya, yang tercermin dalam dokumen
Anggaran Pendapatan Belanja Negara (APBN) untuk nasional dan Anggaran
Pendapatan Belanja Daerah (APBD) untuk daerah atau regional. 15http://ekaputralamury.blogspot.co.id/2012/12/perbedaan-belanja-pegawai-pada-belanja.html,
akses
tanggal 21 januari 2016, pukul: 11.04
16 muslimpoliticians.blogspot.co.id/2011/10/pengertian-pengertian-pada-struktur.html, akses tanggal 12
januari 2016, pukul 11.43
17 Moh.Arsjad Anwar dkk, Ekonomi Indonesia, Masalah dan prosfek 1989/1990, (Jakarta: Universitas
Indonesia Press, 1989), hlm. 50
18 Sadono Sukirno, Ekonomi Pembangunan, Edisi Kedua, (Jakarta: Prenadamedia Group, 2006), hlm. 9
19 Sjahrir, Analisis Ekonomi Indonesia, (Jakarta: Gramedia Pustaka Utama, 1991), hlm. 50 1. Investasi Tujuan dari
kebijakan fiskal ini adalah dalam rangka menstabilkan harga, tingkat output,
maupun kesempatan kerja dan memacu atau mendorong pertumbuhan
ekonomi.13 Pengeluaran pemerintah adalah pengeluaran yang sifatnya habis dipakai
dalam proses produksi setelah dikurangi dengan penjumlahan barang dan jasa
yang dihasilkan oleh pemerintah, yang terdiri dari pembelian barang dan jasa
(belanja barang), pembayaran balas jasa pegawai (belanja pegawai), dan
penyusutan barang modal, dikurangi dengan hasil penjualan barang dan jasa
(output pasar) pemerintah yang tidak dipisahkan dari kegiatan pemerintah.14 Adapun komponen pengeluaran pemerintah ini terbagi atas belanja
langsung dan belanja tidak langsung, Belanja langsung adalah belanja yang
terkait dengan produktivitas kegiatan atau langsung dengan tujuan organisasi, A m b o k P a n g i u k - P e n g a r u h I n v e s t a s i | 8 8 misalnya belanja pegawai, honor yang merupakan sesuatu yang harus
dibayarkan oleh pemerintah kepada pegawai tetapi apabila pegawai tidak
melakukan pekerjaannya maka upah tidak akan dibayarkan.15 Berdasarkan Peraturan Menteri Dalam Negeri Nomor 13 Tahun 2006
Tentang Pedoman Pengelolaan Keuangan Daerah, mengenai belanja langsung
yang terdapat dalam Pasal 50, Kelompok belanja langsung dari suatu kegiatan
dibagi menurut jenis belanja yang terdiri dari belanja pegawai, belanja barang
dan jasa serta belanja modal.16 Belanja tidak langsung adalah belanja yang
secara tidak langsung terkait dengan produktivitas atau tujuan organisasi. pengeluaran tidak langsung meliputi: Belanja pegawai, belanja barang, subsidi
daerah otonom, pembayaran bunga dan cicilan hutang serta pengeluaran rutin
lainnya.17 19 Sjahrir, Analisis Ekonomi Indonesia, (Jakarta: Gramedia Pustaka Utama, 1991), hlm. 50 ,
),
18 Sadono Sukirno, Ekonomi Pembangunan, Edisi Kedua, (Jakarta: Prenadamedia Group, 2006), hlm. 9 3.
Pertumbuhan Ekonomi Pertumbuhan ekonomi adalah suatu ukuran kuantitatif yang
menggambarkan perkembangan suatu perekonomian dalam suatu tahun
tertentu apabila dibandingkan dengan tahun sebelumnya.18 Untuk menghitung
tingkat pertumbuhan ekonomi akan selalu digunakan rumus: g =
x 100
GDP1-GDP0
GDP0 g =
x 100
GDP1-GDP0
GDP0 Keterangan: Keterangan: g adalah tingkat (persentase) pertumbuhan ekonomi g adalah tingkat (persentase) pertumbuhan ekonomi
GDP1 adalah pendapatan nasional riil yaitu pendapatan nasional yang dihitung
pada harga tetap yang dicapai dalam suatu tahun
GDP0 adalah pendapatan nasional riil pada tahun sebelumnya.19 g adalah tingkat (persentase) pertumbuhan ekonomi
GDP1 adalah pendapatan nasional riil yaitu pendapatan nasional yang dihitung
pada harga tetap yang dicapai dalam suatu tahun GDP0 adalah pendapatan nasional riil pada tahun sebelumnya.19 9 | Iltizam Journal Of Shariah Economic Research, Vol. 1, No. 1, 2017 HASIL PENELITIAN DAN PEMBAHASAN Penelitian ini diukur menggunakan metode penelitian kuantitatif deskriptif,
yang secara deskriptif peneliti menjelaskan fenomena atau fakta yang ditemui
penulis dengan cara menggambarkan data sekunder mengenai kondisi investasi,
pengeluaran pemerintah dan pertumbuhan ekonomi Provinsi Jambi pada tahun
yang bersangkutan dengan bantuan tabel, grafik ataupun diagram. Sedangkan secara kuantitatif peneliti akan melakukan analisis statistik
dengan menggunakan metode regresi linier berganda, pengujian statistik dan uji
asumsi klasik yang akan menganalisis data sekunder yang telah diperoleh dari
sumbernya dengan menggunakan bantuan perangkat lunak (Software). Dengan
adanya pengujian ini akan diperoleh hasil mengenai pengaruh dan besaran
pengaruh antara variabel investasi dan pengeluaran pemerintah terhadap variabel
pertumbuhan ekonomi Provinsi Jambi. Setelah dilakukan pengolahan data dengan menggunakan bantuan perangkat
lunak (softwere) maka diperoleh hasil analisis regresi linier berganda sebagai berikut : Hasil Analisis Regresi Linier Berganda Hasil Analisis Regresi Linier Berganda
Variable
Coefficient
Std. Error
t-Statistic
Prob. C
7.416.681
0.032554
2.278.241
0.0000
LOG(INV?)
0.095354
0.017309
5.509.091
0.0000
LOG(GE?)
0.003171
0.001414
2.243.193
0.0330
Sumber: Data diolah Dari hasil output menggunakan softwere tersebut diperoleh persamaan sebagai
berikut: Log Y= Log 7.416.681 + Log 0.095354 * Inv + Log 0.003171 * GE Log Y= Log 7.416.681 + Log 0.095354 * Inv + Log 0.003171 * GE Nilai koefisien dari masing-masing variabel dapat diartikan sebagai berikut : 1. Nilai konstanta (a) sebesar 7.416.681, artinya jika Investasi dan pengeluaran
pemerintah = 0 maka pertumbuhan ekonomi sebesar 7.416.681, dengan asumsi
cateris paribus. 1. Nilai konstanta (a) sebesar 7.416.681, artinya jika Investasi dan pengeluaran
pemerintah = 0 maka pertumbuhan ekonomi sebesar 7.416.681, dengan asumsi
cateris paribus. A m b o k P a n g i u k - P e n g a r u h I n v e s t a s i | 10 10 2. Nilai koefisien elastisitas investasi sebesar 9,53 % yang memberikan arti bahwa
setiap kenaikan nilai investasi sebesar 1 % akan direspon oleh peningkatan
pertumbuhan ekonomi sebesar 9,53 % dengan asumsi cateris paribus. Hasil ini
sesuai dengan penelitian sebelumnya yang dilakukan oleh Efrizal hasan, Syamsul
anwar dan Ali anis dalam jurnal yang berjudul pengaruh investasi, tenaga kerja
dan pengeluaran pemerintah terhadap pertumbuhan ekonomi Provinsi Sumatra
Barat. 2. Nilai koefisien elastisitas investasi sebesar 9,53 % yang memberikan arti bahwa
setiap kenaikan nilai investasi sebesar 1 % akan direspon oleh peningkatan
pertumbuhan ekonomi sebesar 9,53 % dengan asumsi cateris paribus. Hasil ini
sesuai dengan penelitian sebelumnya yang dilakukan oleh Efrizal hasan, Syamsul
anwar dan Ali anis dalam jurnal yang berjudul pengaruh investasi, tenaga kerja
dan pengeluaran pemerintah terhadap pertumbuhan ekonomi Provinsi Sumatra
Barat. 3. Nilai koefisien elastisitas pengeluaran pemerintah sebesar 0,31 % yang
memberikan arti bahwa setiap kenaikan nilai pengeluaran pemerintah sebesar 1
% akan direspon oleh peningkatan pertumbuhan ekonomi sebesar 0,31 %
dengan asumsi cateris paribus. Hasil ini sesuai dengan temuan penelitian yang
dilakukan oleh Sayekti Suindyah D dalam jurnalnya yang berjudul pengaruh
investasi, tenaga kerja dan pengeluaran pemerintah terhadap pertumbuhan
ekonomi Provinsi Jawa Timur. Adapun untuk analisis dari masing-masing kabupaten/kota diperolah
output sebagai berikut: Analisis Regresi Linier Berganda Perkabupaten
Fixed Effects (Cross)
_KERINCI—C
0.005550
_MERANGIN—C
0.001824
_SAROLANGUN—C
0.001071
_BATANGHARI—C
0.000573
_MUARAJAMBI—C
-0.001264
_TANJABTIMUR—C
-0.001600
_TANJABBARAT—C
-0.005707
_TEBO—C
0.001790
_BUNGO—C
4.86E-05
_KOTAJAMBI—C
-0.002286
Sumber: Data diolah Analisis Regresi Linier Berganda Perkabupaten Analisis Regresi Linier Berganda Perkabupaten Dari tabel tersebut dapat diketahui bahwa Investasi dan pengeluaran
pemerintah
memiliki
pengaruh
terhadap
pertumbuhan
ekonomi
perkabupaten/kota diProvinsi Jambi dengan besaran pengaruh yang berbeda-beda. 11 | Iltizam Journal Of Shariah Economic Research, Vol. 1, No. 1, 2017 1. Nilai koefisien elastisitas untuk kabupaten Kerinci adalah sebesar 0,005550
artinya jika Investasi dan pengeluaran pemerintah = 0 maka pertumbuhan
ekonomi kabupaten Kerinciakan naik sebesar 0,55 %, dengan asumsi cateris
paribus. Log Y= Log 7.416.681 + Log 0.095354 * Inv + Log 0.003171 * GE 2. Nilai koefisien elastisitas untuk kabupaten Merangin adalah sebesar 0,001824
artinya jika Investasi dan pengeluaran pemerintah = 0 maka pertumbuhan
ekonomi kabupaten Meranginakan naik sebesar 0,18 %, dengan asumsi cateris
paribus. 2. Nilai koefisien elastisitas untuk kabupaten Merangin adalah sebesar 0,001824
artinya jika Investasi dan pengeluaran pemerintah = 0 maka pertumbuhan
ekonomi kabupaten Meranginakan naik sebesar 0,18 %, dengan asumsi cateris
paribus. 3. Nilai koefisien elastisitas untuk kabupaten Sarolangun adalah sebesar 0,001071
artinya jika Investasi dan pengeluaran pemerintah = 0 maka pertumbuhan
ekonomi kabupaten Sarolangunakan naik sebesar 0,10 %, dengan asumsi cateris
paribus. 4. Nilai koefisien elastisitas untuk kabupaten Batanghari adalah sebesar 0,000573
artinya jika Investasi dan pengeluaran pemerintah = 0 maka pertumbuhan
ekonomi kabupaten Batanghari akan naik sebesar 0,05 %, dengan asumsi cateris
paribus. 5. Nilai koefisien elastisitas untuk kabupaten Muaro Jambi adalah sebesar -
0,001264 artinya jika Investasi dan pengeluaran pemerintah = 0 maka
pertumbuhan ekonomi kabupaten Muaro Jambi akan turun sebesar 0,12 %,
dengan asumsi cateris paribus. 6. Nilai koefisien elastisitas untuk kabupaten Tanjab Timur adalah sebesar -
0,001600 artinya jika Investasi dan pengeluaran pemerintah = 0 maka
pertumbuhan ekonomi kabupaten Tanjab Timur akan turun sebesar 0,16 %,
dengan asumsi cateris paribus. 7. Nilai koefisien elastisitas untuk kabupaten Tanjab Barat adalah sebesar -
0,005707 artinya jika Investasi dan pengeluaran pemerintah = 0 maka
pertumbuhan ekonomi kabupaten Tanjab Barat akan turun sebesar 0,57 %,
dengan asumsi cateris paribus. 7. Nilai koefisien elastisitas untuk kabupaten Tanjab Barat adalah sebesar -
0,005707 artinya jika Investasi dan pengeluaran pemerintah = 0 maka
pertumbuhan ekonomi kabupaten Tanjab Barat akan turun sebesar 0,57 %,
dengan asumsi cateris paribus. A m b o k P a n g i u k - P e n g a r u h I n v e s t a s i | 12 12 8. Nilai koefisien elastisitas untuk kabupaten Tebo adalah sebesar 0,001790
artinya jika Investasi dan pengeluaran pemerintah = 0 maka pertumbuhan
ekonomi kabupaten Tebo akan naik sebesar 0,17 %, dengan asumsi cateris
paribus. 9. Nilai koefisien elastisitas untuk kabupaten Bungo adalah sebesar 4,8605 artinya
jika Investasi dan pengeluaran pemerintah = 0 maka pertumbuhan ekonomi
kabupaten Bungo akan naik sebesar 4,86 %, dengan asumsi cateris paribus. 9. Nilai koefisien elastisitas untuk kabupaten Bungo adalah sebesar 4,8605 artinya
jika Investasi dan pengeluaran pemerintah = 0 maka pertumbuhan ekonomi
kabupaten Bungo akan naik sebesar 4,86 %, dengan asumsi cateris paribus. 10. 1. Uji Normalitas Uji normalitas digunakan untuk mengetahui apakah dalam penelitian
berdistribusi normal atau tidak.20Dasar pengambilan keputusannya adalah nilai
Jarque Bera (JB) yang diperoleh harus lebih kecil dari nilai X2tabel. Uji normalitas digunakan untuk mengetahui apakah dalam penelitian
berdistribusi normal atau tidak.20Dasar pengambilan keputusannya adalah nilai
Jarque Bera (JB) yang diperoleh harus lebih kecil dari nilai X2tabel. Setelah dilakukan pengujian pada data, maka diperoleh nilai Jarque Bera (JB)
yang lebih kecil dari nilai X2tabel (0,293 < 5,991), maka dapat disimpulkan bahwa
residual berdistribusi normal. (JB) Setelah dilakukan pengujian pada data, maka diperoleh nilai Jarque Bera (JB)
yang lebih kecil dari nilai X2tabel (0,293 < 5,991), maka dapat disimpulkan bahwa
residual berdistribusi normal. 20 Steffi Sigilipu, “Pengaruh Penerapan Informasi Akuntansi Manajemen dan Sistem Pengukuran Kinerja Terhadap
Kinerja Manajerial”, Jurnal EMBA Vol. 1 No. 3 (Juni 2013), hlm. 244 Log Y= Log 7.416.681 + Log 0.095354 * Inv + Log 0.003171 * GE Fixed Effect lebih
baik dari Random
Effect
Sumber: Data diolah Perbandingan Model
Pooled OLS
Fixed Effect
Random Effect
F hitung= 247,065
F hitung = 481,688
Hausman
Test(X2hitung)
=
65,147
F tabel 5% = 3,32
F tabel 5% = 3,32
X2tabel = 5,991
F hitung > F tabel
maka signifikan
F hitung > F tabel
maka signifikan
X2hitung
>
X2tabel
maka
signifikan
Hasil uji Chow test antara Pooled OLS dengan
Fixed Effect : F hitung = 481,688
F tabel 5% = 3,32
F hitung > F tabel maka signifikan artinya
estimasi model dengan menggunakan fixed effect
lebih baik dibandingkan dengan pooled OLS. Fixed Effect lebih
baik dari Random
Effect
Sumber: Data diolah Perbandingan Model Perbandingan Model Setelah dilakukan uji asumsi klasik terhadap data-data yang telah
dikumpulkan maka diperoleh hasil output sebagai berikut: 1. Uji Normalitas 1. Uji Normalitas Log Y= Log 7.416.681 + Log 0.095354 * Inv + Log 0.003171 * GE Nilai koefisien elastisitas untuk Kota Jambi adalah sebesar -0,002286 artinya
jika Investasi dan pengeluaran pemerintah = 0 maka pertumbuhan ekonomi
Kota Jambi akan turun sebesar 0,22 %, dengan asumsi cateris paribus. 10. Nilai koefisien elastisitas untuk Kota Jambi adalah sebesar -0,002286 artinya
jika Investasi dan pengeluaran pemerintah = 0 maka pertumbuhan ekonomi
Kota Jambi akan turun sebesar 0,22 %, dengan asumsi cateris paribus. Setelah dilakukan pengolahan data dari 10 kabupaten/kota tersebut dapat
diketahui bahwa pertumbuhan ekonomi kabupaten/kota yang paling tinggi
dipengaruhi oleh investasi dan pengeluaran pemerintah adalah kabupaten Tanjung
Jabung Barat, Kota Jambi, Kabupaten Tanjung Jabung Timur dan Kabupaten
Muaro Jambi. Dalam penelitian dengan menggunakan data panel, harus dilakukan uji
Hausman yang bertujuan untuk mengetahui apakah model pengaruh tetap atau
model pengaruh acak. Statistik hasil uji Hausman dapat dilihat pada tabel berikut: Hasil Uji Hausman
Model
Chi-Square
Chi-Square Tabel 5%
df=2
Prob. Fixed dan Random
65,147
5,991
0,0000
Sumber: Data diolah Hasil Uji Hausman
Model
Chi-Square
Chi-Square Tabel 5%
df=2
Prob. Fixed dan Random
65,147
5,991
0,0000
Sumber: Data diolah Hasil uji Hausman memperoleh nilai X2hitung > X2tabel (65,147 >5,991)
pada taraf signifikansi 5% atau nilai –p hasil uji Hausman sebesar 0,000 < 0,05. Sehingga dapat dinyatakan bahwa model yang menggambarkan pengaruh investasi
dan pengeluaran pemerintah terhadap pertumbuhan ekonomi adalah model tetap
(Fixed). Hasil uji mengindikasikan bahwa analisis data panel dengan pengaruh tetap 13 | Iltizam Journal Of Shariah Economic Research, Vol. 1, No. 1, 2017 13 lebih baik dibanding dengan model pengaruh acak. Adapun hasil uji perbandingan
ketiga model tersebut adalah sebagai berikut: Perbandingan Model
Pooled OLS
Fixed Effect
Random Effect
F hitung= 247,065
F hitung = 481,688
Hausman
Test(X2hitung)
=
65,147
F tabel 5% = 3,32
F tabel 5% = 3,32
X2tabel = 5,991
F hitung > F tabel
maka signifikan
F hitung > F tabel
maka signifikan
X2hitung
>
X2tabel
maka
signifikan
Hasil uji Chow test antara Pooled OLS dengan
Fixed Effect : F hitung = 481,688
F tabel 5% = 3,32
F hitung > F tabel maka signifikan artinya
estimasi model dengan menggunakan fixed effect
lebih baik dibandingkan dengan pooled OLS. a. Persamaan Regresi: Log Y= Log a0 + Log * Inv + Log * GE b. Estimasi Regresi : b. Estimasi Regresi : X1 = b0+ Log * GE
X2 = b0+ Log * INV X1 = b0+ Log * GE
X2 = b0+ Log * INV Untuk persamaan (1) nilai R2 adalah sebesar 0,4925 selanjutnya disebut R2
1
Untuk persamaan (2) nilai R2 adalah sebesar 0,9900 selanjutnya disebut R2
11
Untuk persamaan (3) nilai R2 adalah sebesar 0,9895 selanjutnya disebut R2
12
Diketahui bahwa nilai R2
1< R2
11 , R2
12 (0,4925 < 0,9900, 0,9895) artinya model
ini tidak mengandung gejala multikolinieritas. 4. Uji Autokorelasi 2. Uji Heteroskedastisitas Pada uji Heteroskedastisitas diperoleh hasil p value obs*- Square atau biasa
disebut X2tabelyang lebih besar daria. Yaitu 5,991 > 0,05, artinya dengan
tingkat
keyakinan
95%
model
regresi
ini
dikatakan
bebas
gejala A m b o k P a n g i u k - P e n g a r u h I n v e s t a s i | 14 14 heteroskedastisitas. Dengan demikian pada data investasi dan pengeluaran
pemerintah tidak terdapat gejala heteroskedastisitas. 3. Uji Multikolinieritas 3. Uji Multikolinieritas 3. Uji Multikolinieritas Tahapan pengujian dengan pendekatan korelasi parsial dengan tahapan
sebagai berikut : a. Persamaan Regresi: a. Persamaan Regresi: 4. Uji Autokorelasi Uji ini untuk melihat terjadinya anggota-anggota dari serangkaian
pengamatan yang tersusun dalam rangkaian waktu. Uji autokorelasi dilihat
melalui nilai Durbin-Watson. Nilai DW yang diperoleh adalah sebesar 1,
776.Nilai du dengan k = 2 dan n = 40 adalah 1,600, sedangkan nilai 4-du (4-
1,600) adalah 2,400, sehingga klasifikasi nilai DW berada pada interval yaitu
1,600 < 1,776 < 2,400 hal inimenunjukkan bahwa pada model regresi ini tidak
terdapat gejala autokorelasi. Setelah dilakukan uji statistik terhadap data penelitian, maka diperoleh
hasil sebagai berikut: 1. Koefisien Determinasi (R2) 1. Koefisien Determinasi (R2) Koefisien determinasi (R2) bertujuan mengukur seberapa jauh kemampuan
model dalam menerangkan variasi variabel terikat. Dalam penelitian ini koefisien 15 | Iltizam Journal Of Shariah Economic Research, Vol. 1, No. 1, 2017 15 determinasi untuk mengukur seberapa jauh kemampuan variabel bebas (investasi
dan pengeluaran pemerintah) menjelaskan variabel terikat (pertumbuhan
ekonomi Provinsi Jambi). Nilai Koefisien determinasi (R2) dari hasil regresi linier berganda adalah
sebesar 0,492. Artinya 49,2 % keragaman investasi dan pengeluaran pemerintah
mempengaruhi keragaman tingkat pertumbuhan ekonomi Provinsi Jambi. Sedangkan sisanya sebesar 50,8 % dipengaruhi oleh keragaman lain, yang tidak
disebutkan dalam model ini. 2. Uji Parsial (Uji T) Uji parsial dengan alpha = 5 % dan derajat kebebasan n-k-1 (40-2-1)
diperoleh t tabel sebesar 2,042. Dari hasil perhitungan dengan regresi linier
berganda untuk pengujian pengaruh investasi terhadap pertumbuhan ekonomi
diperoleh nilai koefisien regresi parsial = 0,095354 sedangkan standar error
diperoleh = 0,017309, sehingga diperoleh nilai t hitung sebesar 5,509 dengan
p=0,000. Nilai t tabel pada taraf signifikansi 5 % adalah 2,042. Dikarenakan
nilai t hitung > t tabel (5,509 > 2,042) artinya Investasi secara parsial
berpengaruh signifikan terhadap pertumbuhan ekonomi Provinsi Jambi. Sedangkan dari hasil perhitungan regresi linier berganda untuk pengujian
pengaruh pengeluaran pemerintah terhadap pertumbuhan ekonomi diperoleh
nilai koefisien regresi parsial = 0,003171 sedangkan standar error diperoleh =
0,001414, sehingga diperoleh nilai t hitung sebesar 2,243 dengan p= 0,033. Nilai t tabel pada taraf signifikansi 5 % adalah 2,042. Dikarenakan nilai t hitung
> t tabel (2,243 > 2,024) artinya pengeluaran pemerintah secara
parsial
berpengaruh signifikan terhadap pertumbuhan ekonomi Provinsi Jambi. . Uji Simultan (Uji F) 3. Uji Simultan (Uji F) Uji simultan dengan alpha = 5 % dan derajat kebebasan (k, N-k-1) maka
(2, 40-2-1) sehingga diperoleh F tabel sebesar 3,32. Dari hasil perhitungan diperoleh nilai F hitung sebesar 247,065 dengan
p=0,026. Dikarekan F hitung > F tabel (247,065 > 3,32) artinya investasi dan A m b o k P a n g i u k - P e n g a r u h I n v e s t a s i | 16 16 pengeluaran pemerintah secara bersama-sama (simultan) berpengaruh terhadap
pertumbuhan ekonomi Provinsi Jambi. Setelah dilakukan pengujian pada data yang dikumpulkan maka
diperoleh hasil bahwa dalam pengujian secara parsial investasi sebagai variabel X1
berpengaruh signifikan terhadap pertumbuhan ekonomi hal ini menunjukkan
bahwa investasi yang selalu meningkat tiap tahunnya telah mampu menopang
pertumbuhan ekonomi, akan tetapi investasi di Provinsi Jambi masih harus
ditingkatkan karena pada data investasi perkabupaten diprovinsi Jambi terlihat
penanaman investasi yang dilakukan belum merata keseluruh kabupaten,
penanaman modal yang dilakukan investor hanya ada pada beberapa kabupaten
seperti Tanjung Jabung Barat, Muaro Jambi dan Muaro Bungo sementara pada
kabupaten Kerinci dan Muaro Tebo investasi terlihat belum memadai. Begitupula dengan pengeluaran pemerintah pada 9 kabupaten dan 1 kota
di Provinsi Jambi yang setiap tahun mengalami peningkatan, hasil yang diperoleh
adalah pengeluaran pemerintah berpengaruh positif yang signifikan terhadap
pertumbuhan ekonomi Provinsi Jambi, artinya pengeluaran pemerintah telah dapat
berkontribusi terhadap kondisi pertumbuhan ekonomi. Pengeluaran pemerintah
yang selalu berusaha ditingkatkan pemerintah dengan tetap memperhatikan aspek
efisiensinya, yang berupa belanja langsung dapat memicu meningkatnya
kesejahteraan masyarakat, salah satunya dengan pembayaran gaji pegawai dengan
demikian pendapatan perkapita masyarakat akan meningkat dan hal ini akan
meningkatkan pertumbuhan ekonomi. KESIMPULAN Setelah melakukan uji asumsi klasik maupun uji statistik dengan menggunakan
metode regresi linier berganda dan menganalisis data sekunder mengenai pengaruh
investasi dan pengeluaran pemerintah terhadap pertumbuhan ekonomi Provinsi
Jambi dari tahun 2012-2015, maka dapat disimpulkan bahwa : 17 | Iltizam Journal Of Shariah Economic Research, Vol. 1, No. 1, 2017 a). Pengaruh Investasi terhadap Pertumbuhan Ekonomi Provinsi Jambi. a). Pengaruh Investasi terhadap Pertumbuhan Ekonomi Provinsi Jambi. Analisis mengenai pengaruh investasi terhadap pertumbuhan ekonomi Provinsi
Jambi menunjukkan hasil yang positif signifikan, sebagaimana ditunjukkan
pada besaran pengaruh yang ditimbulkan oleh variabel Investasi, Nilai koefisien
elastisitasnya sebesar 0,0953 jika Investasi mengalami kenaikan 1 % maka
pertumbuhan ekonomi yang ditunjukkan dengan PDRB berdasarkan harga
konstan tahun 2000 akan naik sebesar 9,53 % dengan asumsi cateris paribus atau
variabel yang lain dianggap tetap. b).Pengaruh pengeluaran pemerintah terhadap Pertumbuhan Ekonomi Provinsi
Jambi dalam analisis mengenai pengaruh pengeluaran pemerintah terhadap
pertumbuhan ekonomi Provinsi Jambi menunjukkan hasil yang positif
signifikan. Diperoleh hasil koefisien elastisitas sebesar 0,031, artinya jika
pengeluaran pemerintah mengalami kenaikan 1 % maka pertumbuhan ekonomi
akan naik sebesar 0,31 % dengan asumsi cateris paribus atau variabel yang lain
dianggap tetap. c).Pengaruh Investasi dan Pengeluaran pemerintah terhadap pertumbuhan
ekonomi Provinsi Jambi. Dari hasil analisis yang dilakukan diperoleh nilai
konstanta sebesar 7.416.681 artinya, jika investasi dan pengeluaran pemerintah
= 0 maka tingkat pertumbuhan ekonomi yang ditunjukkan dengan PDRB atas
dasar harga konstan tahun 2000 adalah sebesar 7.416.681,- dengan asumsi
cateris paribus atau variabel lain dianggap tetap. Setelah dilakukan uji f (uji
simultan) diperoleh nilai signifikan 0,026 yang lebih kecil dari taraf keyakinan
0,05, yang artinya investasi dan pengeluaran pemerintah secara simultan
(bersama-sama)
berpengaruh signifikan terhadap pertumbuhan ekonomi
Provinsi Jambi. c).Pengaruh Investasi dan Pengeluaran pemerintah terhadap pertumbuhan
ekonomi Provinsi Jambi. Dari hasil analisis yang dilakukan diperoleh nilai
konstanta sebesar 7.416.681 artinya, jika investasi dan pengeluaran pemerintah d).R square yang diperoleh adalah sebesar 0,492 yang berarti bahwa 49,2%
keragaman investasi dan Pengeluaran pemerintah mempengaruhi keragaman
tingkat pertumbuhan ekonomi Provinsi Jambi. Sedangkan sisanya sebesar 50,8
% dipengaruhi oleh keragaman lain, yang tidak disebutkan dalam model ini. A m b o k P a n g i u k - P e n g a r u h I n v e s t a s i | 18 18 DAFTAR PUSTAKA Alim dan Budi Sutrisno, Hukum Investasi di Indonesia, Jakarta: Raja Grafindo
Persada, 2008 Ana Rokhmatussa’dyah & Suratman, Hukum Investasi & Pasar Modal, Jakarta: Sinar
Grafika, 2011 Adi Raharjo, “Pengaruh Pengeluaran Pemerintah, investasi swasta dan angkatan kerja
terhadap pertumbuhan ekonomi tahun 1982-2003 (studi kasus dikota Semarang)”,
Tesis Universitas Diponegoro Semarang, (2006) Anton Hermanto Gunawan, Anggaran Pemerintah dan Inflasi di Indonesia, Jakarta:
Gramedia Pustaka Utama, 1991 Arsjad Anwar dkk, Ekonomi Indonesia, Masalah dan prosfek 1989/1990, Jakarta:
Universitas Indonesia Press, 1989 Badan Pusat Statistik Provinsi Jambi, Analisis Pertumbuhan Ekonomi Provinsi Jambi
2013, Jambi: BPS Provinsi Jambi, 2013 2013, Jambi: BPS Provinsi Jambi, 2013
Bambang Juanda, Ekonometrika Pemodelan dan Pendugaan, Bogor: IPB Press, 2009
Burhan Bungin, Metode Penelitian Sosial dan Ekonomi, Jakarta: Kencana, 2013
BPS Provinsi Jambi, Jambi dalam angka Tahun 2012, Jambi: BPS Provinsi Jambi,
2012
BPS, Statistik Daerah Provinsi Jambi 2013, Jambi, BPS Provinsi Jambi, 2013
BPS Provinsi Jambi, Jambi dalam angka Tahun 2014, Jambi: BPS Provinsi Jambi,
2014
BPS Provinsi Jambi, Analisis Pertumbuhan Ekonomi Provinsi Jambi 2014, Jambi, BPS
Provinsi Jambi, 2014
Djamaries, Kamus lengkap 10 Milyar, Jakarta: Citra Harta Prima
Digilib.uinsuska.ac.id/8212/1/BAB%201,%20V,%20DAFTAR%PUSTAKA.pdf. Akses tanggal 09 Nopember 2015, pukul 14.35
Efrizal Hasan, Syamsul Amar, Ali Anis, “Pengaruh Investasi, Angakatan kerja dan
Pengeluaran Pemerintah terhadap Pertumbuhan Ekonomi diProvinsi Sumatra Barat”. Bambang Juanda, Ekonometrika Pemodelan dan Pendugaan, Bogor: IPB Press, 2009
Burhan Bungin, Metode Penelitian Sosial dan Ekonomi, Jakarta: Kencana, 2013 Hasan, Syamsul Amar, Ali Anis, “Pengaruh Investasi, Angakatan kerja dan
engeluaran Pemerintah terhadap Pertumbuhan Ekonomi diProvinsi Sumatra Barat”. 19 | Iltizam Journal Of Shariah Economic Research, Vol. 1, No. 1, 2017 Harry A. P. Sitaniapessy, “Pengaruh pengeluaran pemerintah terhadap PDRB dan PAD”,
Jurnal Economia, volume 9, nomor 1, (April 2013) Harry A. P. Sitaniapessy, “Pengaruh pengeluaran pemerintah terhadap PDRB dan PAD”,
Jurnal Economia, volume 9, nomor 1, (April 2013) http://ekaputralamury.blogspot.co.id/2012/12/perbedaan-belanja-pegawai-pada-
belanja.html, akses tanggal 21 januari 2016, pukul: 11.04 http://www.stiualhikmah.ac.id/index.php/kecerdasan-finansial/188-investasi-
dalam-pandangan-al-qur-an-sunnah, akses tanggal 11 06 2016, pukul 14.34
Jhingan, Ekonomi Pembangunan dan Perencanaan, Jakarta: Rajawali Press, 2012
Lailahamkha.blogspot.co.id, Akses tanggal 9 juni 2016, pukul 05.43 Murdifin Haming & Salim Basalamah, Studi Kelayakan Investasi Proyek &
Bisnis,Jakarta: Bumi Aksara, 2010 Michael P. Todaro & Stephen C. Smith, Pembangunan Ekonomi, Edisi Kesembilan,
Indonesia, PT. Gelora Aksara Pratama, 2006 Mohammad Rizal Mubaroq dkk, “Pengaruh Investasi Pemerintah, Tenaga Kerja, dan
Desentralisasi Fiskal Terhadap Pertumbuhan Ekonomi Kabupaten Di Indonesia Tahun
2007 – 2010”, Jurnal M. DAFTAR PUSTAKA Rizal M Met UNPAD, (Januari 2013) Muslimpoliticians.blogspot.co.id/2011/10/pengertian-pengertian-pada-
struktur.html, akses tanggal 12 januari 2016, pukul 11.43. Peraturan Daerah Provinsi Jambi Nomor 2 Tahun 2009 Peraturan Daerah Provinsi Jambi Nomor 2 Tahun 2009 J
Richard T. Gill, Ekonomi Pembangunan Dulu dan Sekarang, Jakarta: Ghalia Indonesia,
1983
Rosmawati Sinuraya, “Analisis Pengaruh Pengeluaran Pemerintah terhadap pertumbuhan
ekonomi kabupaten Karo”, Skripsi Universitas Sumatra Utara, (2010)
Rencana Pembangunan Jangka Panjang Daerah Provinsi Jambi Tahun 2005-2025
Rencana Pembangunan Jangka Menengah Provinsi Jambi tahun 2011-2015
Sadono Sukirno, Ekonomi Pembangunan, Edisi Kedua, Jakarta: Prenada Media
Group, 2006
Sadono Sukirno, Makro Ekonomi Modern, Jakarta: Raja Grafindo Persada, 2007
Sadono Sukirno, Pengantar Ekonomi Makro, Jakarta: Graha Media, 2010
Sayid Syekh, Pengantar Makro Ekonomi, Jakarta: Gaung Persada Press Group, 2014 Richard T. Gill, Ekonomi Pembangunan Dulu dan Sekarang, Jakarta: Ghalia Indonesia,
1983 1983 Rencana Pembangunan Jangka Panjang Daerah Provinsi Jambi Tahun 2005-2025
Rencana Pembangunan Jangka Menengah Provinsi Jambi tahun 2011-2015 Rencana Pembangunan Jangka Menengah Provinsi Jambi tahun 2011-2015 A m b o k P a n g i u k - P e n g a r u h I n v e s t a s i | 20 20 Sayid Syekh, Sekilas Pengantar Ilmu Ekonomi dan Pengantar Ekonomi Islam, Jakarta: GP
Press Group 2013 Press Group, 2013
Sjahrir, Analisis Ekonomi Indonesia, Jakarta: Gramedia Pustaka Utama, 1991
Sayekti Suindyah D, “Pengaruh Investasi, tenaga Kerja dan Pengeluaran pemerintah terhadap
pertumbuhan ekonomi diprovinsi jawa timur, 2009”, Tesis Universitas Darul ‘Ulum
Jombang (2009) Steffi Sigilipu, “Pengaruh Penerapan Informasi Akuntansi Manajemen dan Sistem
Pengukuran Kinerja Terhadap Kinerja Manajerial”, Jurnal EMBA Vol. 1 No. 3
(Juni 2013) Tim Penyusun, Pedoman Penulisan Skripsi, Jambi: Syariah press, 2014 Unit Statistik, Survey dan Liaison, Statistik Ekonomi Keuangan Daerah, Jambi: Bank
Indonesia, 2015 Usman Husaini & Purnomo Setiady akbar, Pengantar Statistika, Edisi Kedua Jakarta:
Bumi Aksara, 2012 W. Riawan Tjandra, Hukum Keuangan Negara, Jakarta: Grasindo, 2006.
|
https://openalex.org/W2570896964
|
https://link.springer.com/content/pdf/10.1007%2Fs10694-016-0645-8.pdf
|
English
| null |
Increasing the Simulation Performance of Large-Scale Evacuations Using Parallel Computing Techniques Based on Domain Decomposition
|
Fire technology
| 2,017
|
cc-by
| 20,376
|
Keywords: Parallel computing, Evacuation, Evacuation simulation, Real-time, Large-scale Keywords: Parallel computing, Evacuation, Evacuation simulation, Real-time, Large-scale * Correspondence should be addressed to: E. R. Galea, E-mail: e.r.galea@gre.ac.uk Increasing the Simulation Performance
of Large-Scale Evacuations Using Parallel
Computing Techniques Based on Domain
Decomposition A. Grandison, Y. Cavanagh, P. J. Lawrence and E. R. Galea*, Fire Safety
Engineering Group, University of Greenwich, London SE10 9LS, UK A. Grandison, Y. Cavanagh, P. J. Lawrence and E. R. Galea*, Fire Safety
Engineering Group, University of Greenwich, London SE10 9LS, UK Fire Technology, 53, 1399–1438, 2017
2017 The Author(s). This article is published with open access at Springerlink.com.
Manufactured in The United States
DOI: 10.1007/s10694-016-0645-8 Fire Technology, 53, 1399–1438, 2017
2017 The Author(s). This article is published with open access at Springerlink.com. Manufactured in The United States
DOI: 10.1007/s10694-016-0645-8 Fire Technology, 53, 1399–1438, 2017
2017 The Author(s). This article is published with open access at Springerlink.com. Manufactured in The United States
DOI: 10.1007/s10694-016-0645-8 Fire Technology, 53, 1399–1438, 2017 Received: 4 May 2016/Accepted: 23 December 2016 Abstract. Evacuation simulation has the potential to be used as part of a decision
support system during large-scale incidents to provide advice to incident comman-
ders. To be viable in these applications, it is essential that the simulation can run
many times faster than real time. Parallel processing is a method of reducing run
times for very large computational simulations by distributing the workload amongst
a number of processors. This paper presents the development of a parallel version of
the rule based evacuation simulation software buildingEXODUS using domain
decomposition. Four Case Studies (CS) were tested using a cluster, consisting of 10
Intel Core 2 Duo (dual core) 3.16 GHz CPUs. CS-1 involved an idealised large
geometry, with 20 exits, intended to illustrate the peak computational speed up per-
formance of the parallel implementation, the population consisted of 100,000 agents;
the peak computational speedup (PCS) was 14.6 and the peak real-time speedup
(PRTS) was 4.0. CS-2 was a long area with a single exit area with a population of
100,000 agents; the PCS was 13.2 and the PRTS was 17.2. CS-3 was a 50 storey high
rise building with a population of 8000/16,000 agents; the PCS was 2.48/4.49 and the
PRTS was 17.9/12.9. CS-4 is a large realistic urban area with 60,000/120,000 agents;
the PCS was 5.3/6.89 and the PRTS was 5.31/3.0. This type of computational perfor-
mance opens evacuation simulation to a range of new innovative application areas
such as real-time incident support, dynamic signage in smart buildings and virtual
training environments. 1. Introduction The smoke and
fire detector data identifies the most closely matching scenarios from a large data-
base of precomputed fire simulations. The GETAWAY system relies on the build-
ingEXODUS evacuation simulation software [9–12] to run evacuation simulations
representing all the alternative evacuation strategies and identify the best route
out of the structure given the evolving situation. Such a system is potentially lim-
ited by the size of the scenarios that can be simulated and the time required to
perform not one, but many simulations. While there are many considerations
which must be taken into account when applying such models to live incidents,
one of the first concerns the speed of computation. In this case the insight that
could be obtained from an evacuation model is only useful if that information can
be used to affect the ongoing incident. Thus for this type of application to be use-
ful, it is essential that the simulation can be run many times faster than real time. Faster runtimes are also useful in the commercial environment where the mod-
elling can be performed more quickly or to a greater level of detail than would be
possible using a conventional non-parallel version of the software. In addition to
faster runtimes the parallel implementation also allows the possibility of running
larger problems than was previously possible. Evacuation simulation tools capable
of simulating large scale events have been used to forensically analyse past tra-
gedies such as the Love Parade in 2010 [13, 14] and the Hajj in 1990 [15, 16] and
2015 [17]. However, the large scale of these events, both in terms of area and
number of people, limits the level of detail that can be represented within these
evacuation simulations. The use of the parallel implementation not only results in
faster runtimes but also enables the possibility of running larger problems at a
higher level of detail than previously possible due to the extra memory available
across the processing units. Similarly, the greater speed of computation and
greater detail that can be achieved using a parallel implementation make these
tools better suited to the planning of large-scale events [18]. Parallel computing techniques are one way of reducing run times for very large
computational simulations. Other ways of reducing runtime can be achieved by
using macroscopic models, hybrid models [19], improved algorithms and faster
hardware. 1. Introduction The use of evacuation/pedestrian modelling is well established as part of building
design to ensure that buildings meet performance based safety and comfort crite-
ria [1, 2]. Another possible use for these models is to provide data, in addition to
sensor information [3, 4], to a decision support system that in turn provides live
operational advice to incident commanders while disasters are actually unfolding
[5]. Live decision support systems that have been suggested include: the FireGrid 1 1 1400 Fire Technology 2017 Fire Technology 2017 [6, 7] system that was designed to aid emergency responders using a combination
of artificial intelligence, sensor data, and predictive computation; the EU FP7
GETAWAY project [8] developed a system where the emergency signage within a
complex building can adapt to a developing hazardous environment and direct
occupants to their optimal exit point using information from smoke and fire
detectors, live CCTV streams, and fire and evacuation simulation. The smoke and
fire detector data identifies the most closely matching scenarios from a large data-
base of precomputed fire simulations. The GETAWAY system relies on the build-
ingEXODUS evacuation simulation software [9–12] to run evacuation simulations
representing all the alternative evacuation strategies and identify the best route
out of the structure given the evolving situation. Such a system is potentially lim-
ited by the size of the scenarios that can be simulated and the time required to
perform not one, but many simulations. While there are many considerations
which must be taken into account when applying such models to live incidents,
one of the first concerns the speed of computation. In this case the insight that
could be obtained from an evacuation model is only useful if that information can
be used to affect the ongoing incident. Thus for this type of application to be use-
ful, it is essential that the simulation can be run many times faster than real time. [6, 7] system that was designed to aid emergency responders using a combination
of artificial intelligence, sensor data, and predictive computation; the EU FP7
GETAWAY project [8] developed a system where the emergency signage within a
complex building can adapt to a developing hazardous environment and direct
occupants to their optimal exit point using information from smoke and fire
detectors, live CCTV streams, and fire and evacuation simulation. 1. Introduction A typical parallel implementation distributes a simulation across a
number of computers making use of the available memory and processing capabil-
ities of all the computers in the cluster or network. There are two main benefits
associated with this approach. Firstly, larger scenarios can be modelled than
would be possible with an evacuation model running on a single computer. This
allows the simulation of very large building complexes or potentially even large
urban spaces to be modelled in far greater detail than was previously possible. Secondly, large scenarios can run more quickly than was previously possible and
potentially much faster than real time. 1401 Increasing the Simulation Performance of Large-Scale Evacuations Increasing the Simulation Performance of Large-Scale Evacuations Most pedestrian evacuation models can be described as macroscopic or micro-
scopic although some models can be described as mesoscopic [20–22]. The macro-
scopic approach (also known as coarse network models [1, 2, 23]) do not
represent individuals but treat the population more like a fluid. They can repre-
sent very large scale evacuations and compute them in short timeframes on a sin-
gle PC without the need for parallel computation. However, macroscopic models
cannot easily represent detailed human behaviour and the interaction of individu-
als [21], e.g. it is difficult to examine contra-flow. The alternative microscopic
approach models the crowd as a collection of interacting agents and so potentially
has the ability to represent and predict interactive behaviours. There are three
commonly used approaches to representing agent behaviour within microscopic
evacuation models: Cellular Automata (CA) [24], Social Forces based models [25–
27], and rule based models [9–12, 28, 29]. CA methods have been parallelised
across multiple processors [30] but CA methods are particularly well suited to par-
allelisation on a SIMD (Single Instruction Multiple Data) platform [31] such as a
General Purpose Graphical Processing Unit (GPGPU) [30, 32, 33] or a Field Pro-
grammable Gate Array (FPGA) [32] due to each grid cell being computed in the
same fashion but with different data. However if complex interactions/behaviours
are to be represented involving agents utilising information obtained from beyond
their immediate vicinity the effectiveness of SIMD parallelism will be affected. Parallelisation can also be applied to social forces based models [34, 35]. In evacu-
ation models utilising the Social Forces approach the movement of agents is gov-
erned by equations representing virtual forces within a continuous spatial domain. 1. Introduction At the boundary of the domain partitions, each sub-domain must com-
municate with its neighbouring sub-domain to transfer agents from one sub-do-
main (and therefore computer) to another sub-domain (computer) as they move
through the environment. Using this strategy, many computers will potentially be
responsible for handling the movement of a particular agent through the environ-
ment. This was the strategy adopted in other parallel implementations [30, 32–35]. In this work a domain decomposition approach is applied to the rule based
evacuation simulation software buildingEXODUS [9–12]. The parallel implemen-
tation is implemented on distributed memory cluster computer networks and a
series of tests evacuation simulations are used to gauge the real time and compu-
tational speedup achieved. 1. Introduction Social Forces models appear to be very computationally intensive and so can
potentially greatly benefit from a parallel implementation. For example, Steffen
et al. [35] noted that an evacuation simulation of a stadium involving 20,000 occu-
pants using a parallel implementation of a Social Forces based evacuation model
could be performed approximately five times faster than real time using a parallel
computer utilising 180 processors. The work presented in this paper differs from the earlier work on parallelisation
of evacuation models by exploring the parallel implementation of a rule based
evacuation model. Given the differences in the formulation of rule based evacua-
tion models and the computational costs associated with rule based models com-
pared with CA and Social Force models, it is unclear how effective parallelisation
can be for this type of model. Furthermore, most of the earlier parallel implemen-
tations were simply concerned with reducing the execution time of the simulation
[30, 33, 34] but not necessarily as part of a live incident, where significantly faster
than real time performance is required. Two implementations were specifically
concerned with running faster than real time for use in a live incident [32, 35]. The Giitsidis et al. [32] model ran on specialist hardware (FPGA) is reported to
achieve run times over 107 times faster than real time when applied to a small air-
craft carrying 150 passengers. It is unclear, from the available literature, if this
model could be extended to simulating larger environments. Steffen et al. [35]
demonstrated their parallel social force evacuation model on a sports stadium. The simulation of the stadium evacuation involving 20,000 pedestrians could run 1402 Fire Technology 2017 Fire Technology 2017 approximately in real-time when 24 processors were used and when 180 processors
were used could run approximately five times faster than real time. Before evacuation simulation could be considered useful for real time applica-
tions, sufficient speedup over real time must be achieved in order to allow incident
managers sufficient time to assess the information and suggest potential mitigation
strategies. Even if the management system is automated, sufficient time must be
available to implement the alternative strategies and for the alternative strategies
to be effective. Thus it is suggested that speedups of at least an order of magni-
tude (10+ times) over real time would be required before evacuation simulation
could begin to be considered useful in real time applications. 1. Introduction There are two potential strategies for parallelisation of the evacuation simula-
tion; these are population decomposition and domain decomposition. Population decomposition sub-divides the population with each processing
thread responsible for a particular population sub-group i.e. an individual will
always be simulated by the same processing thread. This approach is potentially
attractive on a single multi-core/multi-CPU shared memory (SM) based PC using
OpenMP [36] or multithreading. However, extending this approach to a dis-
tributed set of computers (i.e. a cluster) has a number of shortcomings. Each com-
puter must accommodate the entire geometry which would restrict the maximum
size geometry that could be simulated no matter how many computers were used. Also there is potentially a large amount of interaction between the agents simu-
lated on different computers which may incur additional code complexity and high
communication costs between the computers. p
Domain decomposition is based on a systematic partitioning of the problem
domain (geometry) onto a number of sub-domains (sub-geometries) and is the
method generally used in parallelising Computational Fluid Dynamics (CFD)
based simulations including fire simulations [37–40]. Each sub-domain is com-
puted on a separate processor and runs its own copy of the evacuation simulation
software. At the boundary of the domain partitions, each sub-domain must com-
municate with its neighbouring sub-domain to transfer agents from one sub-do-
main (and therefore computer) to another sub-domain (computer) as they move
through the environment. Using this strategy, many computers will potentially be
responsible for handling the movement of a particular agent through the environ-
ment. This was the strategy adopted in other parallel implementations [30, 32–35]. In this work a domain decomposition approach is applied to the rule based
evacuation simulation software buildingEXODUS [9–12]. The parallel implemen-
tation is implemented on distributed memory cluster computer networks and a
series of tests evacuation simulations are used to gauge the real time and compu-
tational speedup achieved. Domain decomposition is based on a systematic partitioning of the problem
domain (geometry) onto a number of sub-domains (sub-geometries) and is the
method generally used in parallelising Computational Fluid Dynamics (CFD)
based simulations including fire simulations [37–40]. Each sub-domain is com-
puted on a separate processor and runs its own copy of the evacuation simulation
software. 2. Parallel Implementation The implementation effort required for an advanced egress model can be substan-
tial before considering the parallel processing. It was therefore decided to base the
parallel implementation on an existing advanced egress code, buildingEXODUS. Increasing the Simulation Performance of Large-Scale Evacuations 1403 buildingEXODUS is a well validated software product and the source code was
readily available to the authors. The buildingEXODUS software has been described many times in the literature
[9–12] and so only a brief description is provided here. EXODUS is a suite of
software tools designed to simulate the evacuation of large numbers of people
from complex enclosures. The software is a multi-agent simulation environment
that utilises a two-dimensional grid of nodes to represent space. The software
takes into consideration people–people, people-fire and people-structure interac-
tions. The model tracks the trajectory of each individual as they make their way
out of the enclosure, or are overcome by fire hazards such as heat, smoke and
toxic gases. The behaviour and movement of each individual agent is determined
by a set of heuristics or rules. The spatial grid maps out the geometry of the
building, locating exits, internal compartments, obstacles, etc. The grid is made up
of nodes and arcs with each node representing a small region of space and each
arc representing the distance between each node. Individuals travel from node to
node along the arcs. In creating a parallel implementation of an existing serial code that has a wide
international user base there are a number of core requirements that the parallel
implementation must satisfy in order to make the parallel version of the software
both flexible and easy to adopt by the existing user base. These include the follow-
ing considerations: There should be no difference between the input or output files for the serial
and parallel implementations of the software. This will allow applications to be
designed and the results visualised using the familiar serial components of the
software. The parallel implementation should work on any number of processors. Minimal additional investment (in time and hardware/software) should be
required by the engineer to effectively run the software. The system should
work on a conventional set of PCs attached via a standard LAN as well as a
dedicated cluster. The parallel implementation is intended to function in a Microsoft Windows
environment. 2. Parallel Implementation An additional requirement imposed by the software developers was that there
should only be one EXODUS source code i.e. separate parallel and serial source
codes would not be developed. Having a single software source code is desirable
in order to minimise the effort required to maintain the product. Thus the parallel
code was developed by modifying the existing serial source code. There are a number of parallel processing technologies available that include
multi-core CPUs, networked computers (e.g. specialised clusters or general office
networked computers), GPGPU and FPGA. Although GPGPU and FPGA coding looks attractive for evacuation simulation
[32], due to their impressive performance compared to CPUs, it is difficult to
apply to a rule based evacuation model where each agent will follow its own
branching path of execution within the software making it unsuitable, at present, 1404 Fire Technology 2017 Fire Technology 2017 for a SIMD type processor [31] which can only perform a single execution branch. Another issue is the complexity of the buildingEXODUS software which has hun-
dreds of thousands lines of code making writing a GPGPU or FPGA version a
major undertaking even if it was suitable. buildingEXODUS software is suitable for a Multiple Instruction Multiple Data
(MIMD) [31] style of parallel processing as MIMD systems are able to simultane-
ously execute multiple execution pathways. Both multi-core CPUs and networked
computers are MIMD systems. MIMD systems are able to simulate different
branches of execution which is required for the rule-based movement used within
buildingEXODUS. The de-facto Application Programming Interface (API) for distributed parallel
processing is MPI (Message Passing Interface) [41] and the version created by
Argonne National Laboratory, MPICH2 [42] for Windows, was used to facilitate
the parallel implementation. Using this API reduces the amount of programming
effort required to make a parallel implementation that works on both multi-core
CPUs and networked computers. 2.1. Software Parallelisation Strategy Domain decomposition was the chosen parallelisation strategy for this implemen-
tation as it favours the Distributed Memory (DM) model of parallel processing
and was the strategy adopted in other parallel implementations [30, 32–35]. In PC
parlance this is a network (or cluster) of PCs linked together via some form of
Local Area Network (LAN). It should be further noted that the program written
for a DM system would also run on a SM machine removing the need for two
separate parallel versions. The decomposition concept is illustrated in Fig. 1 where
the domain is simply split into two sub-domains; however the domain can be split
into many sub-domains. The code was designed using the Single Program Multiple Data (SPMD) paradigm. With this methodology only one executable is used and a copy of the executable is
launched on each processor which operates on its own part of the problem domain. Typically there is one process that handles the I/O and problem distribution, this is
the master process (process 0). The other processes are exactly the same as one
another and only differ slightly from the master; the master process was also responsi-
ble for visualisation and collection of the results to a single output file. 2.2. Agent Transfer and Movement Across Sub-domain Boundaries In creating the sub-domains a series of Halo nodes are added to each partition
boundary of each sub-domain (see Fig. 1). Essentially, Halo nodes are an addi-
tional series of nodes that are added to the partition boundaries of a sub-domain
that copy the information stored in the boundary nodes of the adjoining sub-do-
mains. We refer to the additional series of nodes added to the partition boundary
as Outer Halo nodes, while the series of nodes within the sub-domain on the par-
tition boundary are called Inner Halo nodes. The Halo nodes are used to pass
information between neighbouring sub-domains concerning the movement of indi-
viduals between the neighbouring sub-domains as illustrated in Fig. 2. Here the 1405 Increasing the Simulation Performance of Large-Scale Evacuations Inner Halo /
Boundary nodes
Outer Halo
Inner halo of one sub-geometry is mapped
to the outer halo of adjoining sub-geometry
Partition geometry
into sub-geometries
and add halo nodes
Figure 1. Example decomposition of a computational domain into
two sub-domains. 2.1. Software Parallelisation Strategy Inner halo of one sub-geometry is mapped
to the outer halo of adjoining sub-geometry Inner halo of one sub-geometry is mapped
to the outer halo of adjoining sub-geometry Inner Halo /
Boundary nodes Boundary nodes
Outer Halo Figure 1.
Example decomposition of a computational domain into
two sub-domains. (b) Agent enters Inner Halo
(d) Agent now moves on
neighbouring sub-domain (a) Agent approaches sub-domain boundary
(c) Agent moves to Outer Halo (b) (d) Agent now moves on
neighbouring sub-domain Figure 2. The movement of an agent across sub-domains in the par-
allel version of EXODUS. Figure 1.
Example decomposition of a computational domain into
two sub-domains. Figure 1. Example decomposition of a computational domain into
two sub-domains. domain has been split into two sub-domains with each sub-domain being placed
on a separate computer. Each computer is responsible for performing the compu-
tations for the evacuation simulation for the agents who are located on their
assigned sub-domain. In the example shown in Fig. 2, the agent is moving from
left to right and must transfer from one sub-domain to the other. The movement
across sub-domain boundaries can be briefly explained as follows: (a) In Fig. 2a, the agent is approaching the sub-domain boundary but is currently
controlled by the left hand sub-domain (LHSD). (a) In Fig. 2a, the agent is approaching the sub-domain boundary but is currently
controlled by the left hand sub-domain (LHSD). (b) As the agent enters the Inner Halo region of the LHSD (see Fig. 2b), the
LHSD now sends a message concerning the agent stood on the Inner Halo
node to the right hand sub-domain (RHSD) and the RHSD now creates a
copy of the agent on its Outer halo node. (b) As the agent enters the Inner Halo region of the LHSD (see Fig. 2b), the
LHSD now sends a message concerning the agent stood on the Inner Halo
node to the right hand sub-domain (RHSD) and the RHSD now creates a
copy of the agent on its Outer halo node. (c) As the agent moves onto the Outer Halo region of the LHSD (see Fig. 2c) this
information is sent to the RHSD which updates the agent on its side to move
to its Inner Halo region. At this point the agent is now controlled by the
RHSD. (d) In Fig. 2d, the agent now continues its journey on the RHSD and as it moves
away from the boundary the LHSD is instructed to delete its copy of the
agent as that copy is no longer needed. (d) In Fig. 2d, the agent now continues its journey on the RHSD and as it moves
away from the boundary the LHSD is instructed to delete its copy of the
agent as that copy is no longer needed. 1406 Fire Technology 2017 (b) Agent enters Inner Halo
(a) Agent approaches sub-domain boundary
(c) Agent moves to Outer Halo
(d) Agent now moves on
neighbouring sub-domain
Figure 2. The movement of an agent across sub-domains in the par-
allel version of EXODUS. Figure 2.
The movement of an agent across sub-domains in the par-
allel version of EXODUS. 2.2.1. Synchronisation In the above explanation the communication model has
been simplified. It is not the case that one computer can simply send a message to
another. The communication needs to be arranged to ensure that the appropriate
computer is ready to receive a message and the corresponding computer is sending
the message. Failure to correctly orchestrate this communication can lead to dead-
lock. This occurs when both computers are waiting to receive a message from
each other but they will wait forever as the message will not be sent until a mes-
sage is received from the other computer. This was solved by using a convention
based on the sub-domain numbering and using partial non-blocking communica-
tion. All sub-domains are uniquely labelled 0 to N-1, where N is the total number
of sub-domains. This leads to the general case that each sub-domain will have
boundaries with both lower and higher numbered sub-geometries. These boundaries can be classified as low (neighbouring sub-domain has a
lower number) and high boundaries (neighbouring sub-domain has a higher num-
ber). In order to remain synchronised every sub-domain initiates a ‘receive’ from
its low boundaries and simultaneously ‘sends’ update messages to its high bound-
aries. All boundaries wait until they have successfully received a message from
their low boundaries. Once this has been achieved the computers initiates a ‘re-
ceive’ from the high boundaries while simultaneously sending update messages to
the low boundaries. 2.2.2. Parallel Movement Algorithm Within EXODUS the overall simulation is
governed by the global Simulation Clock which ticks every 1/12 s. On even ticks
agents decide what their future actions will be; this is generally a movement 1407 Increasing the Simulation Performance of Large-Scale Evacuations towards the nearest exit, on odd ticks the movement is performed [9]. The popula-
tion is looped over until all the PETs (Personal Elapsed Time) of each of the
agents has incremented beyond the Simulation Clock due to a movement to the
next node or in congested flows a decision to wait in its current position until the
next tick of the global Simulation Clock. The global Simulation Clock keeps tick-
ing until the scenario finishes. The algorithm for the parallel implementation is essentially the same as that for
the serial implementation except that the population is looped over in sub-groups
related to their proximity to sub-domain boundaries. 1. Simulation Clock is incremented 1/12 s and the ticker is incremented by one
tick. Figure 2.
The movement of an agent across sub-domains in the par-
allel version of EXODUS. These are the high-boundary
group, the low-boundary group, and the non-boundary group. The boundary
groups are further sub-divided into communication groups and computation
groups. A communication group consists of agents that are on the inter sub-do-
main boundary and consists of agents on the inner and outer halo nodes of a sub-
domain. A computation group consists of agents on the inner halo nodes and the
neighbouring set of nodes within the sub-domain. The non-boundary computation
group consists of all the agents that are not members of a boundary computation
group within the sub-domain. This arrangement ensures that agents in the non-
boundary group cannot move directly into the high/low boundary communication
groups when computation and communication are overlapped in steps 3 and 5 in
the algorithm below. In Fig. 3, 4, 5 6 and 7 the computation groups are repre-
sented by a dashed line box, a sending communication group is represented by a
continuous line box, and a receiving communication group is represented by a
dash-dot line box. Below is the logic used in the parallel movement algorithm, with the additional
steps required by the parallel algorithm highlighted in italics (steps 3–7). 1. Simulation Clock is incremented 1/12 s and the ticker is incremented by one
tick. Figure 3. Example of communication and computation in step 3 of
parallel movement algorithm. Figure 3. Example of communication and computation in step 3 of
parallel movement algorithm. 1408 Fire Technology 2017 2. If the number of simulation ticks is even then movement options for individu
als are calculated then go to 1. Else if the number of simulation ticks is od
Figure 4. Example of computation in step 4 of parallel movement
algorithm. Figure 5. Example of communication and computation in step 5 of
parallel movement algorithm. Figure 6. Example of computation in step 6 of parallel movement
algorithm. Figure 4. Example of computation in step 4 of parallel movement
algorithm. Figure 5. Example of communication and computation in step 5 of
parallel movement algorithm. Figure 4. Example of computation in step 4 of parallel movement
algorithm. 2. If the number of simulation ticks is even then movement options for individ
als are calculated then go to 1. Else if the number of simulation ticks is od
then the selected movement will be performed in steps 3–7, continue. Figure 5. Figure 2.
The movement of an agent across sub-domains in the par-
allel version of EXODUS. In the case of agent C that moved into
the high-boundary communication group a new copy of agent C is created on the
j + 1 sub-domain. Agent B in the non-boundary group was computed whilst the
communication was taking place. 4. The agents located around the low boundaries of a sub-domain are computed for
potential movement. In Fig. 4 agent D is seen to move away from the boundary
and will no longer be part of the low-boundary communication group but will be
part of the low-boundary computation group. Agent E crosses over to sub-domain
j and leaves the low-boundary computation group of sub-domain (j + 1) although
it is still a member of the low-boundary communication group of sub-domain
(j + 1). 5. The agents on low boundaries are sent to neighbouring sub-domains whilst at the
same time the agents are received from high boundaries. Whilst the computers
wait to receive/send data from one another, further members of the non-boundary
group are computed for potential movement until this communication is complete. In Fig. 5 agent D’s movement is communicated to the high-boundary communica-
tion group of sub-domain (j - 1) and is deleted from sub-domain (j - 1). Agent
E’s position is communicated from sub-domain (j + 1) to sub-domain j and its
positon is updated on sub-domain j. Agent G and F’s movements in the non-
boundary group are performed during the communication. 6. The agents located around the high boundaries of a sub-domain are computed for
potential movement. In Fig. 6 agent H moves from the high-boundary computa-
tion group across the boundary onto the outer halo nodes of sub-domain (j - 1). 6. The agents located around the high boundaries of a sub-domain are computed for
potential movement. In Fig. 6 agent H moves from the high-boundary computa-
tion group across the boundary onto the outer halo nodes of sub-domain (j - 1). 6. The agents located around the high boundaries of a sub-domain are computed for
potential movement. In Fig. 6 agent H moves from the high-boundary computa-
tion group across the boundary onto the outer halo nodes of sub-domain (j - 1). g
p
y
f
(j
)
7. Any remaining members of the non-boundary group that were not processed dur-
ing step 3 or step 5 are computed for potential movement. In Fig. 7 it can be seen 7. Figure 2.
The movement of an agent across sub-domains in the par-
allel version of EXODUS. Example of communication and computation in step 5 of
parallel movement algorithm. Figure 6. Example of computation in step 6 of parallel movement
algorithm. Figure 5. Example of communication and computation in step 5 of
parallel movement algorithm. Figure 6. Example of computation in step 6 of parallel movement
algorithm. 2. If the number of simulation ticks is even then movement options for individu-
als are calculated then go to 1. Else if the number of simulation ticks is odd,
then the selected movement will be performed in steps 3–7, continue. 2. If the number of simulation ticks is even then movement options for individu-
als are calculated then go to 1. Else if the number of simulation ticks is odd,
then the selected movement will be performed in steps 3–7, continue. Increasing the Simulation Performance of Large-Scale Evacuations 1409 Figure 7. Example of computation in step 7 of parallel movement
algorithm. Figure 7. Example of computation in step 7 of parallel movement
algorithm. 3. The agents on high boundaries are sent to neighbouring sub-domains whilst at the
same time other agents are received on the low boundaries. Whilst the computers
wait to receive/send data from one another, members of the non-boundary group
are computed for potential movement until this communication is complete. This
is illustrated in Fig. 3. In a previous iteration or time-step agents A and C had
been moved to their locations. In the case of agent A this required an update to
its position in the lower boundary group. In the case of agent C that moved into
the high-boundary communication group a new copy of agent C is created on the
j + 1 sub-domain. Agent B in the non-boundary group was computed whilst the
communication was taking place. 3. The agents on high boundaries are sent to neighbouring sub-domains whilst at the
same time other agents are received on the low boundaries. Whilst the computers
wait to receive/send data from one another, members of the non-boundary group
are computed for potential movement until this communication is complete. This
is illustrated in Fig. 3. In a previous iteration or time-step agents A and C had
been moved to their locations. In the case of agent A this required an update to
its position in the lower boundary group. Figure 2.
The movement of an agent across sub-domains in the par-
allel version of EXODUS. Any remaining members of the non-boundary group that were not processed dur-
ing step 3 or step 5 are computed for potential movement. In Fig. 7 it can be seen 1410 Fire Technology 2017 Fire Technology 2017 that various agents of the non-boundary group are moved. At this point agent H
has not been updated on sub-domain (j). This update will occur when step 3 of
the algorithm is performed again. that various agents of the non-boundary group are moved. At this point agent H
has not been updated on sub-domain (j). This update will occur when step 3 of
the algorithm is performed again. 8. Due to movement conflicts between agents some of the agents will need to be
reprocessed. Go to step 3 until all the remaining agents’ PETs have been incre-
mented. 8. Due to movement conflicts between agents some of the agents will need to be
reprocessed. Go to step 3 until all the remaining agents’ PETs have been incre-
mented. 9. Go to 1 until all the evacuation scenario has finished. 9. Go to 1 until all the evacuation scenario has finished. One of the important developments in the parallel implementation of EXODUS
was the ability to overlap communication with calculation. This overlapping was
critical to obtaining good speedups as without this development the parallel and
thus runtime performance would have been significantly poorer. By overlapping
the communication and computation the order in which agents are processed can
be changed and due to the use of pseudo-random numbers in the software for
some decision making meant that no two runs were the same due to the addi-
tional randomisation now created by the network. This change in ordering is just
as valid as any other particular order but unlike the serial version of EXODUS,
the simulation cannot be repeated to give exactly the same result. An exact repeat
simulation could be obtained by removing the overlapping of communication and
computation at the expense of run time performance; though not generally useful
it was helpful to make the software run in this mode for the purposes of develop-
ment and debugging. 2.3. Decomposition Strategy The decomposition strategy determines how many sub-domains to create and how
the sub-domains should be distributed to the various computers within the cluster/
network. A poor decomposition, or partition, can lead to poor load balancing on
the computers. This means that some of the computers utilised for parallel pro-
cessing are under-utilised and therefore the wall-clock time to run the simulation
is adversely effected. However, unlike CFD fire simulation, where a single domain
decomposition strategy may be appropriate for virtually all fire scenarios, for
evacuation simulation, the decomposition strategy is scenario specific. In the worst
case scenario, at least in terms of parallel performance, it is possible that the par-
allel performance will be no better and possibly worse than the serial performance
no matter how many computers are used if a poor decomposition is selected. Consider a simple example involving a rectangular geometry in which there is
an exit at either end of the geometry and the population is uniformly distributed
throughout the environment. The population is further assumed to be smoothly
flowing with no congestion. If each agent moves towards their nearest exit, half
the population will move to the exit on the left and half will move to the exit on
the right. In this case the ideal partition for a network of two computers would
involve simply splitting the domain in half, with one half associated with the left
exit and the other half associated with the right exit. However if the scenario involved the right exit being non-viable, the simple par-
titioning would be inappropriate as the computer allocated the right part of the 1411 Increasing the Simulation Performance of Large-Scale Evacuations domain would have increasingly less work to do as the population progressively
moves toward the only viable exit located on the left. A point would be reached
when the processor handling the right domain would be idle while the processor
handling the left domain would still be working hard. A simplified analytical calculation of computational performance can be devised
if communication overheads and memory access speed are ignored. It is assumed
that z agents are uniformly distributed in the domain and travel with the same
speed and direction. The length of the domain is l. 2.3. Decomposition Strategy If a single computer was used
to compute the evacuation then the total work performed by that processor is,
(number of agents (z)) 9 (average distance travelled (l/2)), zl/2. In the case of two
processors using the decomposition suggested the calculation is split into 2 parts. Whilst the right hand domain is being emptied, into the left hand sub-domain, the
work performed by the busy processor is, (number of agents on busy processor (z/
2)) 9 (distance travelled (l/2)), zl/4. The second processor is now idle and the
work done in emptying the remaining sub-domain is, (number of agents on busy
processor (z/2)) 9 (average distance travelled (l/4), zl/8. The total work done by
the busy processor is therefore 3zl/8. Computational Speedup (Sp) [43] is defined
as the time taken to run the simulation on one processor (t1) divided by the time
taken to run on P processors/computers (tp) i.e. SP = t1/tp. Assuming that the
work done is proportional to the time taken then the maximum computational
speedup of two processors over one processor for this decomposition would be 4/
3 (1.33). Another difficulty with this simple partition is that it only applies to two pro-
cessors and cannot be generalised for any number of processors. If the population
was not initially uniformly distributed throughout the domain this would also lead
to a computational load imbalance. These problems can be mitigated by using multiple sub-domains per computer
as illustrated in Fig. 8. In this decomposition the diagonally shaded areas are
computed on one processor and the crosshatched shaded areas are computed on
another. Using this scheme the workload is more evenly balanced. As the popula-
tion moves toward the exit both processors are generally kept busy, it is only
when the geometry has emptied to the last sub-domain that the second processor
becomes idle. By generalising the analysis that was performed previously the fol-
lowing analytical computational speedup Eqs. 1 for 2 processors can be obtained
when the domain is split into N equal sized sub-domains, where N is a multiple of
two. SN
2 ¼
2N
N þ 1
ð1Þ SN
2 ¼
2N
N þ 1 SN
2 ¼
2N
N þ 1
ð1Þ ð1Þ So if the domain is split into 6 sub-domains as illustrated in Fig. 2.3. Decomposition Strategy 8 the analytical
speedup is 12/7 (1.71) and as N tends to infinity it can be seen that the analytical
speedup tends to 2. This methodology can be extended to using any number of processors and on
any arbitrary geometry using any decomposition strategy. For the above partition
for two processors the partition can be represented as 121212, if three processors 1412 Fire Technology 2017 Exit
2
1
2
2
1
1 were being used then the partition would be represented as 123123. With this
technique the computational load on each computer is kept well balanced and the
load is more evenly balanced with more partitions per processor. Equation 1 can
be generalised (see Fig. 9) to include any number of processors as well (Eq. 2). Exit
2
1
2
2
1
1
Figure 8. Evacuation domain consisting of rectangular geometry, one
exit and uniform population distribution split into multiple sub-do-
mains and allocated to two processors. Exit were being used then the partition would be represented as 123123. With this
technique the computational load on each computer is kept well balanced and the
load is more evenly balanced with more partitions per processor. Equation 1 can
be generalised (see Fig. 9) to include any number of processors as well (Eq. 2). The derivation of Eq. 2 and Eq. 5 is described in Appendix A. Equation 2 is used
to calculate the analytical computational speedups in case study 2. Figure 8. Evacuation domain consisting of rectangular geometry, one
exit and uniform population distribution split into multiple sub-do-
mains and allocated to two processors. Figure 8. Evacuation domain consisting of rectangular geometry, one
exit and uniform population distribution split into multiple sub-do-
mains and allocated to two processors. were being used then the partition would be represented as 123123. With this
technique the computational load on each computer is kept well balanced and the
load is more evenly balanced with more partitions per processor. Equation 1 can
be generalised (see Fig. 9) to include any number of processors as well (Eq. 2). The derivation of Eq. 2 and Eq. 5 is described in Appendix A. Equation 2 is used
to calculate the analytical computational speedups in case study 2. 2.3. Decomposition Strategy SN
P ¼
N 2P
N N þ P 1
ð
Þ þ KP K2 HðKÞP
ð2Þ SN
P ¼
N 2P
N N þ P 1
ð
Þ þ KP K2 HðKÞP ð2Þ where where K ¼ N mod P
ð3Þ
H K
ð Þ ¼
0;
if K ¼ 0
1;
if K > 0
ð4Þ K ¼ N mod P K ¼ N mod P K ¼ N mod P ð3Þ H K
ð Þ ¼
0;
if K ¼ 0
1;
if K > 0
ð H K
ð Þ ¼
0;
if K ¼ 0
1;
if K > 0
ð4Þ If K = 0 then Eq. 2 simplifies to the following Eq. 5. If K = 0 then Eq. 2 simplifies to the following Eq. 5. If K = 0 then Eq. 2 simplifies to the following Eq. 5. If K = 0 then Eq. 2 simplifies to the following Eq. 5. SN
P ¼
NP
N þ P 1
ð5Þ SN
P ¼
NP
N þ P 1 ð5Þ It can be seen from this particular example (Eq. 5) that speedup improves as the
number of sub-domains increases. Equation 5 is not generally applicable but illus-
trates the advantage of the multiple sub-domains per processor decomposition
strategy. It can be seen from this particular example (Eq. 5) that speedup improves as the
number of sub-domains increases. Equation 5 is not generally applicable but illus-
trates the advantage of the multiple sub-domains per processor decomposition
strategy. However the disadvantage of this technique is the increase in communications
and hence communication costs associated with an increasing number of sub-do-
mains. The increasing cost is due to the increase in the number of sub-domain
boundaries which require agents to be transferred across. This communication has
an overhead and there will be a point where the advantage of better computa- Increasing the Simulation Performance of Large-Scale Evacuations 1413 Exit
P
1
2
N-K-P+1
1
P
1
2
P
K
1
2
N
N-K+1
Figure 9. A generalised domain decomposition for N sub-domains
and P processors where K = N (mod P) and N P for a long area with
a single exit point. P
1
2
N-K-P+1
P
P
K
1
2
N
N-K+1 N Figure 9. 2.3. Decomposition Strategy A generalised domain decomposition for N sub-domains
and P processors where K = N (mod P) and N P for a long area with
a single exit point. tional load balance is outweighed by the extra communication cost. There is
therefore an optimal number of sub-domains which will give a peak speedup per-
formance. This balance between better load balancing that is achieved by having
more sub-domains, the number of processors utilised and the communication
overhead is explored later in case study 2. The general strategy used is to have
multiple sub-domains per processor that are distributed in such a way as to even
out the work load throughout the simulation. 3. Case Studies To demonstrate the parallel implementation four different cases were examined
(see Table 1). It is not the intention of these demonstration cases to provide any
substantive analysis of evacuation behaviour but to demonstrate the performance
improvements possible using the parallel implementation of buildingEXODUS. The first two cases are theoretical in nature and have been designed to gain a
greater understanding of the performance of the parallel implementation. The sec-
ond two cases are based on realistic scenarios; a high-rise building and a large
public gathering in a large urban space. Earlier results produced by a prototype
implementation of the parallel buildingEXODUS [44] have been improved using
the latest version of the parallel implementation described in this paper. The cases demonstrated here have been tested on a 64bit-Windows cluster con-
sisting of 109 Intel Core 2 Duo (dual core) 3.16 GHz based computers connected
via a 1Gbit Ethernet switch. The software is stochastic in nature and needs to be run a number of times,
generating a distribution of predicted evacuation times, which would also result in
a small distribution of runtimes. However, for the purposes of this demonstration
work only, the software was configured to operate in a deterministic manner. Thus, in situations where some movement decisions/conflicts are sometimes deter-
mined by random selection, they have been resolved in a deterministic manner. This was done to eliminate variability between simulations. This would further
ensure that the runtimes of the simulations were consistent. Each case study con-
figuration was run three times and it was found that there was less than a second Fire Technology 2017 1414 Table 1
Case Studies Summary for the Parallel Implementation Table 1
Case Studies Summary for the Parallel Implementation
Case
Population size
Area (m2)
Exits
1. Idealised large geometry
100,000
100,000
20
2. Long open area
100,000
100,000
1
3. 50 Floor high-rise building
8000/16,000
90,000
8
4. Large public area
60,000/120,000
46,000
14 Table 1
Case Studies Summary for the Parallel Implementation Studies Summary for the Parallel Implementation difference in runtime for any particular configuration. The predicted evacuation
time for each case study was identical across the serial and parallel configurations. The results have been split into single core and dual core results. 3. Case Studies It has been
found in previous work that better speedups are obtained on two single processors
connected via a network compared to two processors (cores) on a shared memory
machine [38, 39]; typically dual cores operate at a computational speedup of 1.7–
1.9 compared to a single core. This effect is due to memory bus contention on the
shared memory computer with both processes trying to simultaneously access the
single memory bus within the computer. However, in some cases this memory bus
contention is so high that the use of two processors in a shared memory computer
produces little, if any, speedup compared to a single processor. In the remainder of this paper two types of speedup will be discussed; these are
computational speedup and real-time speedup. Computational speedup compares the
run time of the parallel implementation against the serial or single processor run
time [36]. Real-time speedup, or speed up over real time, compares the run time of
the parallel or serial implementation of the software against the time required to
complete the evacuation as predicted by the software. More detailed tables of
results are available in Appendix B. 3.1. Case 1: Idealised Large Geometry This test case is intended to represent an ideal case for the parallel implementa-
tion. It has been designed so that there is no sub-domain boundary interaction
and that the problem is well load balanced throughout the entire simulation. This
test was devised to explore the upper limits of speedup potential possible with
parallel buildingEXODUS. The geometry is 4000 m long and 25 m wide producing an area of 100,000 m2. There are twenty 5 m wide exit points located along one side of the geometry,
each separated by 200 m with the first and last exits being 100 m from each end
of the geometry. A population of 100,000 agents (1 person/m2) is uniformly dis-
tributed throughout the domain and move towards their nearest exit point. In order to ensure the most optimal decomposition the domain is split into 20
equal sub-domains (each sub-domain has 5000 agents) and these are allocated to
each processor by taking the modulus of the sub-domain number with the total
number of processors used. Due to the layout of the geometry the population
does not have to cross between processor boundaries to reach an exit point. 1415 ncreasing the Simulation Performance of Large-Scale Evacuations Increasing the Simulation Performance of Large-Scale Evacuations The following configurations were used to test the speedup potential: 1, 2, 3, 4,
5, 7 and 10 single cores; 1, 2, 3, 4, 5 and 10 dual cores. Configurations that have
not been represented here would not give any speedup improvements on the con-
figurations that have been simulated. For example with 5 (single core) computers
each computer would have 4 sub-domains to process; if 6 (single core) computers
were utilised then 4 computers would have 3 sub-domains but 2 computers would
have 4 sub-domains and this would be limit the speedup to being the same as 5
(single core) computers. The analytical computational speedup in this instance is
calculated by dividing the total number of sub-domains by the maximum number
of sub-domains residing on any individual processor. The predicted evacuation
time for this scenario was 14 min 26 s and was consistent across all the parallel
simulations and the serial version of buildingEXODUS. The computational
speedup for the various permutations are presented in Fig. 10. An impressive computational speedup of 10.9 was achieved using 10 computers
compared to a single computer. 3.1. Case 1: Idealised Large Geometry An examination of all the single core results
shows that a super-linear speedup, a speedup greater than the number of proces-
sors, is achieved for all the configurations. It is difficult to say precisely why a
super-linear speedup has resulted, but could be due to hardware effects [45, 46],
such as the increase in overall processor cache size. The maximum computational speedup of 13.9 achieved with 10 dual core com-
puters is less than the analytical speedup of 20. The analytical speedup is based on
the premise that doubling the processor/core count would double the speedup. This ignores the inter-processor communication cost and also the effect of cores
sharing the memory bus when the computer is utilising dual cores. These speedups
are good and are unlikely to be achieved for most practical scenarios. They do Figure 10. Analytical and actual computational speedup for the ide-
alised large geometry (Case 1). Figure 10. Analytical and actual computational speedup for the ide-
alised large geometry (Case 1). 1416 Fire Technology 2017 however demonstrate a peak performance that can be achieved from this type of
application. however demonstrate a peak performance that can be achieved from this type of
application. In a real scenario near peak performance will be achieved in the initial stages of
the simulation as there is likely to be a good load balance due to the initial popu-
lation distribution being reasonably uniform. However, as the simulation pro-
gresses it is expected that the parallel efficiency will decrease as the load becomes
unbalanced due to the movement of people. It is also worth noting that on a single processor the simulation was some 3.5
times slower than real time for this particular simulation i.e. on a single processor
the run time was 3.5 times slower than the time required for the actual evacua-
tion. However, using 10 dual core processors produced a real-time speedup of 4. 3.2. Case 2: Long Open Area As the number of processor/cores is increased there is a greater Increasing the Simulation Performance of Large-Scale Evacuations 1417 Figure 11. Single (a) and Dual (b) core actual and analytical compu-
tational Speedup for the Long Open Area (Case 2). Figure 11. Single (a) and Dual (b) core actual and analytical compu-
tational Speedup for the Long Open Area (Case 2). divergence between the analytical speedups and therefore load balances. When 10
dual core computers are used the disadvantage of increased communication for
the 100 sub-domain partition compared to the 20 sub-domain partition is far out-
weighed by the improved load balance achieved. The run time for a single core is 1.2 times faster than the predicted evacuation
time. The real-time speedup for 10 dual core processors using the 100 sub-domain
partition is 17.2. Additional processors should further improve the speedup. 3.2. Case 2: Long Open Area This scenario consists of 100,000 agents in an open rectangular geometry measur-
ing 100 m by 1000 m producing an area of 100,000 m2 resulting in a crowd den-
sity of 1 person/m2. The crowd moves to the left to exit the geometry. The left
side of the geometry is completely open, creating a 100 m wide exit. This case can
be considered as a realisation of the analytical study in Sect. 2.3. The geometry was sliced into N equally sized sub-domains in the same fashion
as illustrated in Fig. 9. For this case the domain was split into 20, 50 and 100
sub-domains respectively. Each of these partitions was examined using 1–10 com-
puters in both single and dual core processor modes and this resulted in 60 total
permutations for this one case. The predicted evacuation time for this geometry is 14 min 20 s. This predicted
time was consistent across the various parallel implementations and with the serial
version of buildingEXODUS. This particular simulation was designed to represent a non-congested exit flow
thus leading to a short overall predicted evacuation time with minimal computa-
tional requirements. The computational speedup for single and dual core configu-
rations are depicted in Fig. 11. The analytical speedup is calculated using Eq. 2. Theoretically 100 sub-partitions should give the best speedup performance for
all the possible permutations due to the best load balance being maintained
throughout the simulation. In practice there are other factors that influence the
performance including the cost of communication, which increases with additional
sub-domains, and hardware effects such as increased cache size and, in the case of
multi-cores, a shared memory bus. The use of a shared memory bus and increas-
ing communication cost both reduce the actual performance from the analytical
prediction. Conversely the increased cache size made available by increasing the
number of computers improves the performance beyond the analytical prediction. The actual speedup performance is therefore a combination of the load balance
and these other factors. When two computers are used, either in single or dual
core mode, the best partition is 20 sub-domains for this particular problem. This
is due to the analytical load balance being comparable to the other partitions but
the communication cost is far lower than the partitions with a higher number of
sub-domains. 3.3. Case 3: High-Rise Building The high-rise building scenario consists of 50 floors with four emergency exit
staircases and a floor area for each floor of 1800 m2. The staircases are not
equally spaced within the building core leading to some of the staircases attracting
more people than others. This test case was run with two population sizes of 8000
agents (160 people per floor) and 16,000 agents (320 people per floor) that were
uniformly distributed throughout the building on each floor. Unlike the previous
case, this scenario involves a great deal of congestion as agents attempt to gain
access to the stairs and as they traverse down the stairs. The predicted evacuation
time for this case was 50 min for 8000 agents and 1 h 30 min for 16,000 agents. The partitioning strategy adopted for this high rise building was based on divid-
ing the building into 25 sub-domains vertically i.e. two floors per sub-domain. These sub-domains are then divided horizontally into four quadrants of equal
floor area with a staircase associated with each quadrant. In total there were 200
sub-domains used in the analysis. The computational speedup obtained for both the single core and dual core
processors are presented in Fig. 12. We note that the computational speedup pro-
duced for the 16,000 agent population is significantly higher than the 8000 popu-
lation. Clearly, the performance of the parallel implementation improves with
increasing problem size. This is a well-known phenomenon with most parallel pro-
cessing problems where the speedup performance improves with increasing prob- 1418 Fire Technology 2017 Fire Technology 2017 lem size [47]. This is due to the relative decrease of communication time with
increasing computation size; this decrease is partly attributable to the fact that
network communications have a fixed start-up cost in addition to the cost of
sending the actual data. For this particular problem it is also related to the fact
that increasing the population size by a factor of two will approximately increase
the communication cost by a factor of two. However, the computational cost has
increased by a factor of 4.5 due to the increased congestion. We also note from Fig. 12 that the performance for the 8000 population has
tailed offas more processors are added to the cluster, with no improvements being
derived from adding more than eight processors to the cluster. 3.3. Case 3: High-Rise Building However, for the
16,000 population further improvements in performance could be derived by add-
ing additional computers (single core) to the cluster beyond 10, but the return
gained from adding additional computers is diminishing. Using 8 single core computers, the evacuation of the 8000 population can be
simulated in 2 min 48 s. The predicted evacuation time is 50 min 7 s. This repre-
sents a real-time speedup of 17.9. The 16,000 population requires 1 h 30 min to
evacuate and using 10 processors the evacuation can be simulated in just over
7 min, representing a real-time speedup factor of 12.9. From Fig. 12 it is noted that the dual core simulations can return poorer per-
formance than the single core simulations. Furthermore, unlike the single core
simulations, the speedup reaches a peak for the 8000 population with 7 processors
and 8 processors for the 16,000 population. Adding additional computers beyond
these critical values actually diminishes performance. As noted earlier, the perfor-
mance of the parallel implementation improves with increasing problem size. As
the dual core computers have a greater computational performance, they require a Figure 12. Actual computational speedup for the high-rise building
(Case 3). Figure 12. Actual computational speedup for the high-rise building
(Case 3). Increasing the Simulation Performance of Large-Scale Evacuations 1419 larger problem size to make better use of the number of computers available in
the cluster. larger problem size to make better use of the number of computers available in
the cluster. 3.4. Case 4: Large Urban Space This case study is a rough approximation of the Trafalgar Square public area in
central
London,
UK. The
usable
pedestrian
area
measures
approximately
46,000 m2 and there are 14 exit routes from the domain (see Fig. 13). Two scenarios were created by populating the domain with 60,000 agents (1.3
p/m2) and 120,000 agents (2.6 p/m2). Unlike the other test cases the agents were
assigned exit points prior to the evacuation leading to more congestion than might
be expected from a more usual scenario where individuals leave via their nearest
exit point. The exit allocation is roughly proportional to the size of each exit and
the individuals allocated to a particular exit are randomly located in the domain. The exit allocation used in this study is not intended to be an accurate representa-
tion of actual behaviour in large urban places but is simply used to test the paral-
lel implementation when agents will be contra-flowing and not simply exiting by
their ‘nearest’ exit. The predicted overall evacuation time for 60,000 people is 10 min 27 s, and for
120,000 people is 21 min 49 s. It should be noted due to the lack of real data and
hypothetical exiting conditions for the individuals that no firm conclusions should
be drawn concerning the evacuation of Trafalgar Square from these simulations. This test case is purely designed to investigate the performance of the parallel
implementation on a large urban space. Figure 13. Example partition for large urban space (Case 4) with
five computers and 20 partitions. Figure 13. Example partition for large urban space (Case 4) with
five computers and 20 partitions. 1420 Fire Technology 2017 Fire Technology 2017 For this case the Metis [48] partitioning algorithm was utilised along with mul-
tiple sub-domains per processor. Metis is generally used for partitioning computa-
tional grids used in Computational Fluid Dynamics (CFD) and Finite Element
Analysis (FEA) for parallel processing. Metis divides the domain into a number
of sub-domains whilst minimizing the size of the sub-domain boundaries. For this
case 10, 20, and 32 sub-domains were created. An example partition is illustrated
in Fig. 13, where each coloured region represents each of 20 sub-domains within
the partition and each coloured sub-domain was allocated to one of five comput-
ers. A visualisation of the evacuation of 60,000 agents is provided in Fig. 3.4. Case 4: Large Urban Space 14 at 3
time slices and the congestion at a number of junctions is clearly shown. The com-
putational speedups for parallel buildingEXODUS on this test case are depicted in
Fig. 15 for a population sizes of 60,000 and 120,000 agents. Results for dual cores
are limited to 10 dual core processors due to time constraints and hardware avail-
ability. While the performance derived from using 20 partitions with single cores
appears erratic it does produce the overall best single core performance for 60,000
agents returning a speedup of 4.73 from 10 processors. Clearly, as in Case 2, the
performance is dependent on the number of partitions used. Using the dual core
processors and 20 partitions a speedup of 5.3 from 10 processors is achieved. It should be noted that the single processor performance is essentially equiva-
lent to real time i.e. it takes as long to compute the evacuation as it does to actu-
ally perform the evacuation. Using 10 single core processors with 20 partitions,
the run time is some 4.8 times faster than real time and runs in just over 2 min. From Fig. 15 we note for the single core simulations, as the number of proces-
sors increases, the speedup also increases. Using 10 dual core processors with 20
partitions, the run time is some 5.31 times faster than real time and runs in just
under 2 min. Using the best partition suggested by the study involving the population of
60,000 individuals the 20 sub-domain case was re-run using a population size of
120,000 agents. (a) Start t = 0 minutes
(b) t =2 minutes
(c) t = 5 minutes
Figure 14. Visualisation of the large public area evacuation (Case 4)
at 0, 2, and 5 min. (c) t = 5 minutes (b) t =2 minutes Figure 14. Visualisation of the large public area evacuation (Case 4)
at 0, 2, and 5 min. Increasing the Simulation Performance of Large-Scale Evacuations 1421 Figure 15. Actual computational speedup graph for the large urban
space (Case 4). Figure 15. Actual computational speedup graph for the large urban
space (Case 4). As is expected, the larger problem size produces a greater computational
speedup. Using single cores, the run time on a single processor is more than twice
as long as the predicted evacuation time. 3.4. Case 4: Large Urban Space However, using 10 single core processors
produces a speedup of 5.94 resulting in 10 single core processors running 2.6 times
faster than real time. Using 10 dual core processors the speedup is a factor of 6.89
on the single core processor time. This results in the 10 dual core processors run-
ning 3 times faster than real time. In Fig. 15 there is an uncharacteristic drop offin performance for nine proces-
sors when 20 sub-domains are used. This is due to the nature of the distribution
of sub-domains on the computers. It is likely that one or two computers are sub-
stantially busier than the rest of the computers due to a poor load balance at
some stage of the simulation. Although this result is notable it can be seen that
the multiple sub-domains per processor is at least partially successful in maintain-
ing speedup on this non-idealised scenario where it is not possible a priori to
determine where the population load balance would exist. 4. Discussion In all cases, the results suggest that better performance, in terms of greater speed-
ups and reduced run times could be achieved by adding additional processors
however; there is a practical limit to the improved performance that can be 1422 Fire Technology 2017 Fire Technology 2017 achieved. This limit in performance is dependent on the complexity of the prob-
lem, the domain decomposition, the problem size, the computing hardware, and
the networking. More complex problems tend to perform poorly compared to
simpler problems. The domain decomposition is a compromise between minimis-
ing communication, e.g. less sub-domains, and increasing load balance, e.g. more
sub-domains. Larger population sizes tend to return better computational perfor-
mance compared to smaller population sizes on the same domain, although smal-
ler population sizes are more likely to return higher real-time speedups. Faster
computing hardware although likely to return improved real-time speedup is likely
to return a poorer computational speedup when compared to slower computing
hardware. More ‘‘distributed’’ processors tend to perform better than the equiva-
lent ‘‘shared’’ processors. Faster networking will improve both computational and
real-time speedups. Examining the cases performed it can be seen that computational speedup gen-
erally improves with larger numbers of agents. This can be seen for case 3 and 4
where that the computational speedup improves particularly for case 3 when the
population size is doubled. For case 3 the load balance should be close to ideal
but is not realised due to the comparatively high communication costs compared
to the amount of computation. When the population is doubled the amount of
computation (time taken) increases by a factor of 4.5 for case 3. This is due to the
increase in congestion that needs to be resolved when there are higher population
densities. The total evacuation time increases by a smaller factor, and therefore
means that although the computational speedup has significantly improved, the
real time speedup will be reduced in this case. For case 4 there is less of an
improvement as the computational speed up highly influenced by the poor load
balance in that instance. Increasing the number of partitions will generally lead to an improved load bal-
ance when the movement of agents is ‘predictable’ as seen in case 2. When the
workload is unpredictable, as in case 4 due to the high level of contra-flow, then
increased partitioning will not always lead to a better load-balance. 4. Discussion From the sin-
gle core results it can be seen that no particular partition is substantively better
than the others. The assignment of sub-domains to processors did not take into
account the relative computational loads on the sub-domains. It was not possible
a priori to determine how to optimally distribute these sub-domains amongst the
processors. In addition to this the size, shape and location of the sub-domains was
not optimised with regard to the load-balance. As the number of processors increases the computational speedup generally
increases although there is a diminishing return with an increased number of pro-
cessors evident for case 3. It is apparent that the computational speedup increases
with the number of processors but there needs to be a sufficient population size to
offset the necessary communications. For case 4 there is a significant dip in com-
putational speedup for 9 processors when 120,000 agents are simulated. As was
previously mentioned the assignment of sub-domains to processors does not take
into account the workload of each sub-domain and has resulted in a particularly
poor load balance in this instance. 1423 Increasing the Simulation Performance of Large-Scale Evacuations Although the strategy of multiple sub-domains per processor has been some-
what successful it can be seen that for case 4 additional work is needed on the
partitioning strategy to improve the computational speedup. Apart from possibly
improving the intitial partition some form of dynamic load balancing is required
that can optimise the workload, per processor, whilst the scenario is running. Dynamic load balancing has been performed in parallel processing for a range of
problems, e.g. fire modelling [39]. The dynamic load balancing could either be per-
formed by: a) reassigning sub-domains from an initially defined partition depen-
dent on population/workload distribution or b) the entire partition could be
recalculated dependent on the evolving population/workload distribution. While the computational speedup is of some interest, the most important con-
sideration is the wall clock time or run time. How long it takes to run an evacua-
tion simulation is one of the key considerations which will determine how
practical it is to use in applications such as an advice tool for incident comman-
ders for large building based scenarios, as part of an interactive emergency sig-
nage system, a training tool in an interactive desk top environment and as a
planning tool for large scale urban applications. ET predicted total evacuation time (m:s), SPRT single processor run time (m:s), BPRT best parallel run time (m:s),
C-SP computational speedup (=SPRT/BPRT), RT-SPS real time speedup (single processor) (=ET/SPRT), RT-SPP
real time speedup (parallel) (=ET/BPRT) 4. Discussion The performance enhancements achieved by the parallel implementation on the
various test cases are summarised in Table 2. As can be seen the performance of
the parallel implementation is complex and dependent on the type of problem
being addressed. Computational speedups (parallel performance over single processor perfor-
mance) ranging from 2.59 to 13.99 were achieved while real-time speedups (par-
allel performance over real-time) of 3.09 to 17.99 were achieved for the various
problem types. Indeed, Case 3—50 floor high-rise building with 8000 agents—re-
turned the worst computational speedup (2.59) but returned the best speedup
over real-time (17.99). (
)
Some of the problems represent very large cases and are extremely computa-
tionally demanding, requiring moderately long real evacuation times with single
processor execution times equal to or exceeding real evacuation times. These are
Case 1 (idealised large geometry case) and the Case 4 (large urban space). For
these problems, it is very difficult to achieve high real-time speedups because the Table 2
Comparison of Runtime Performance of the Four Case Studies
Case
Agents
ET
SPRT
BPRT
C-SP
RT-SPS
RT-SPP
1
100,000
14:26
51:01
3:40
13.9
0.28
3.9
2
100,000
14:20
11:32
0:50
13.8
1.24
17.2
3
8000
50:02
6:57
2:48
2.5
7.2
17.9
3
16,000
90:02
31:26
7:02
4.5
2.9
12.8
4
60,000
10:27
10:25
1:58
5.3
1.0
5.3
4
120,000
21:49
50:13
7:17
6.9
0.43
3.0
ET predicted total evacuation time (m:s), SPRT single processor run time (m:s), BPRT best parallel run time (m:s),
C-SP computational speedup (=SPRT/BPRT), RT-SPS real time speedup (single processor) (=ET/SPRT), RT-SPP
real time speedup (parallel) (=ET/BPRT) ET predicted total evacuation time (m:s), SPRT single processor run time (m:s), BPRT best parallel run time (m:s),
C-SP computational speedup (=SPRT/BPRT), RT-SPS real time speedup (single processor) (=ET/SPRT), RT-SPP
real time speedup (parallel) (=ET/BPRT) 1424 Fire Technology 2017 Fire Technology 2017 single processor performance is already significantly slower than real-time. For
example, in Case 1, the single processor performance is 3.59 slower than real-time
while in Case 4, the single processor performance is 2.39 slower than real time. These cases will produce very good computational speedups (13.99 and 6.99 for
Case 1 and 4 respectively), but relatively poor real-time speedups (3.99 and
5.39 for Case 1 and 4 respectively). 4. Discussion Thus for these applications, the parallel
implementation is very useful for an engineer using the software as part of a
design analysis as it reduces the time required to run the simulations but it is not
very useful for real-time applications which require the software to run many
times faster than real-time. Other problems represent large real cases but are less computationally demand-
ing, requiring very long real evacuation times with single processor execution
times less than real evacuation times. These are Case 3a and Case 3b (high-rise
building with 8000 and 16,000 agents). For these problems, it is easier to achieve
high real-time speedups as the single processor performance is already faster than
real-time. For example, the single processor performance in Case 3a is 2.39 faster
than real-time while in Case 3b it is 2.99 faster than real-time. These cases pro-
duced relatively poor computational speedups (2.59 and 4.59 for Case 3a and 3b
respectively), but relatively good real-time speedups (17.99 and 12.99 for Case 3a
and 3b respectively). Thus these types of cases lend themselves to live applications
where it is essential that computation be performed much faster than real-time e.g. dynamic signage or live incident support. The results obtained from the idealised cases, cases 1 and 2, suggest that much
better speedups could have been obtained for the case 3. Case 2 is of particular
interest as it demonstrates the balance between computation vs communication. Case 2 has a high amount of communication but it does not highly impact the
performance of the parallel implementation on the hardware configuration used
here and is probably due to the successful overlapping of communication and
computation. It also indicates that the relatively poor computational speedups
seen in case 4 are due to poor load balance rather than communication cost. While the parallel implementation has achieved significant performance gains,
further gains in the real-time performance of the buildingEXODUS software are
desirable and indeed achievable. These performance enhancements can be cate-
gorised into hardware and software improvements. Hardware based improvements
include, using faster computers, more computers, and the possibility of using better
networking technology. The networking technology used in the current implementa-
tion utilises standard equipment and as such suffers from comparatively high net-
work latency. The use of specialist network cards and switches could improve the
speedup performance by reducing the latency in the communications. 4. Discussion Improved
data transfer speeds, achieved by for example using high-speed optical fibre cables,
can also enhance the parallel performance by improving the bandwidth between
processors. The main disadvantage of this approach is the comparatively high price
associated with the equipment compared to conventional equipment. Software based improvements include serial algorithmic changes, utilisation of
‘hybrid discretisation’ [19], improved partitioning strategies and dynamic load bal-
ancing. It is possible that further improvements can be made to the serial version Increasing the Simulation Performance of Large-Scale Evacuations 1425 of buildingEXODUS that would increase the runtime performance of the software
which would feed into the parallel implementation of buildingEXODUS. Although this paper has focused on the buildingEXODUS evacuation software
most of the technology could be utilised by other evacuation models. The multiple
sub-domains per processor approach is very generalizable and the derived analyti-
cal expressions could be useful for model developers testing their own software. The concept of halo regions could be used for CA based models and continuous
models. The movement algorithm and the use of boundary and non-boundary
proximity groupings of agents could also be adapted. 5. Conclusion A parallel implementation of the rule based evacuation simulation software build-
ingEXODUS using the technique of domain decomposition has been achieved and
successfully demonstrated in a range of application cases from simple unidirectional
motion to large complex buildings and open spaces. The computational speedups
derived using a 10 processor system over the performance of a single processor var-
ied from 2.59 to 13.99, with real-time speedups varying from 3.09 to 17.99. In all cases examined improved performance, in terms of enhanced speedups and
reduced run-times could be achieved by adding additional processors however; there
is a practical limit to the improved performance that can be achieved. This limit in
performance is dependent on the complexity of the problem, the domain decomposi-
tion, the problem size, the computing hardware, and the networking. The complexity of the problem both in terms of geometry and agent behaviour
will tend to have improved computational speedup with decreasing complexity. Good parallel performance seen with simpler problems may not necessarily be
repeated on more complex problems. The domain decomposition affects the parallel performance, both in terms of
computational speedup and real time speedup, generally improves with the num-
ber of partitions as this generally improves the load balance across processors. However, this can be affected by the complexity of the movement behaviour of
the agents. Where agent movement is not known a priori but evolves due to the
nature of the simulation simply increasing the number of partitions may not nec-
essarily lead to improved speedup. The size of the problem affects the parallel performance, in terms of computa-
tional speedup, which generally increases with the number of agents. However,
real time performance may decrease in situations where the increase in the number
of agents leads to an increase in the population density. The computing hardware will affect the performance. Generally a more ‘‘dis-
tributed’’ computer system will offer a higher parallel performance than an equiv-
alent less distributed system. For example 2 networked single cores provide a
greater performance than a single dual core computer. This is due to the increase
in cache and memory bandwidth offered by the more distributed systems. All
modern CPUs are now multicore but it would be better to have a distributed set
of multicore CPUs rather than a set of multicore CPUs on a single computer. Open Access This article is distributed under the terms of the Creative Commons Attribution
4.0 International License (http://creativecommons.org/licenses/by/4.0/), which per-
mits unrestricted use, distribution, and reproduction in any medium, provided you
give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. 5. Conclusion 1426 Fire Technology 2017 Fire Technology 2017 The networking hardware is significant as communications between sub-do-
mains is a critical factor in determining parallel performance, it is suggested that
the faster the communications between CPUs the better the parallel performance. Thus the faster the networking and the lower the latency within the cluster, the
better will be the parallel performance. Unfortunately, even with good computational speedup the real-time speedup
could be insufficient for real-time applications. In cases where significant real-time
speedups cannot be significantly increased through enhanced hardware (more pro-
cessors, faster processors and faster networking) then other methods will be required
to increase the real-time speedup. This could be achieved by optimising the software
and using hybrid strategies that mix coarse and fine modelling techniques. For innovative evacuation model application areas such as dynamic signage in
smart buildings, real-time incident support and virtual training environments, it is
suggested that real-time speedups of at least an order of magnitude must be
achieved before they can be considered viable. Parallel computing approaches
demonstrated in this paper have generated up to an 18 fold real-time speedup for
some problems. This approach, together with improved algorithms and improved
hardware performance offers a means to address these novel applications. Furthermore, from the cases studied in this paper (particularly case 4), it is sug-
gested that to achieve even better parallel performance requires improving the
load balancing across the processors. This would need to incorporate a dynamic
load balancing algorithm as the work load is continuously varying over time. Acknowledgements The authors acknowledge the UK Home Office CBRN Science and Technology
Programme for financial support enabling the development of the concept for the
parallel implementation of the buildingEXODUS evacuation software. Appendix A: Derivation of Analytical Speedup
performance for Case Study 2 with N sub-domain
and P processors A long area with exiting on the left open side of the domain is considered. It is
assumed that the domain is split into N equally sized sub-domains and are
assigned to P processors as illustrated in Fig. 16. It is assumed that N ‡ P. It is
assumed that all agents travel with the same speed and flow out of the left hand Increasing the Simulation Performance of Large-Scale Evacuations 1427 Exit
P
1
2
N-P
N-2P+1
1
P
1
2
P
i=2
i=N/P
P
1
2
N
N-P+1
i=1
(a)
(a)
Figure 16. A domain decomposition for N sub-domains and P proces-
sors where N mod P = 0. P
1
2
N-P
N-2P+1
i=2
P
1
2
N
N-P+1
i=1
(a)
(a) N P Figure 16. A domain decomposition for N sub-domains and P proces-
sors where N mod P = 0. side with no congestion. The agents are uniformly distributed throughout the
domain. There are assumed to be no communications cost involved in moving the
occupants to the neighbouring sub-domain. side with no congestion. The agents are uniformly distributed throughout the
domain. There are assumed to be no communications cost involved in moving the
occupants to the neighbouring sub-domain. By considering that processor 1 (controlling the solid black sub-domains in the
figure below), will always be running at 100% and assuming the processor
throughput is independent of memory access patterns it is possible to calculate the
amount of work performed by that processor. In this analysis let: l = length of the domain; z = the total initial population size; N = the num-
ber of sub-domains; P = the number of processors. Simplified Derivation In this initial analysis it is assumed that N is an integer multiple of P to simplify
the derivation. Therefore there are N/P regions consisting of P sub-domains label-
led 1 to P. The work performed by processor 1 whilst area (a) is emptied is the number of
agents being computed by processor 1 9 (distance travelled by an agent) this is
represented by:- worka ¼
z
P
l
N
worka ¼
z
P
l
N
ð6Þ ð6Þ worka ¼
P
N
ð6Þ
Work to empty region i = 1 is:
work1 ¼ z l P 1
ð
Þ þ 1
z þ
z z
l
ð7Þ Work to empty region i = 1 is: Work to empty region i = 1 is: Work to empty region i = 1 is: Work to empty region i = 1 is: worki ¼
N
P i 1
ð
Þ
z
N
l
N P 1
ð
Þ
þ 1
2
N
P i 1
ð
Þ
z
N þ
N
P i 1
ð
Þ
z
N z
N
l
N
ð10Þ
worki ¼ zl
N2
N
P i 1
ð
Þ
P 1
ð
Þ þ 1
2
N
P i 1
ð
Þ
þ
N
P i
ð11Þ worki ¼
N
P i 1
ð
Þ
z
N
l
N P 1
ð
Þ
þ 1
2
N
P i 1
ð
Þ
z
N þ
N
P i 1
ð
Þ
z
N z
N
l
N
ð10Þ ð10Þ
þ 1
2
N
P i 1
ð
Þ
z
N þ
N
P i 1
ð
Þ
z
N z
N
l
N
ð10Þ worki ¼ zl
N2
N
P i 1
ð
Þ
P 1
ð
Þ þ 1
2
N
P i 1
ð
Þ
þ
N
P i
ð11 worki ¼ zl
N2
N
P i 1
ð
Þ
P 1
ð
Þ þ 1
2
N
P i 1
ð
Þ
þ
N
P i
ð11Þ ð11Þ Therefore the total work done by processor whilst clearing all regions is: workN; P
total ¼ zl
N 2
X
N=P
i¼1
N
P i 1
ð
Þ
P 1
ð
Þ þ 1
2
N
P i 1
ð
Þ
þ
N
P i
ð12Þ ð12Þ The work needed to empty the domain with a single serial processor (number of
agents 9 average distance travelled) is The work needed to empty the domain with a single serial processor (number of
agents 9 average distance travelled) is workserial
total ¼ zl
2
ð13Þ workserial
total ¼ zl
2
ð13Þ workserial
total ¼ zl
2 ð13Þ The work performed by a processor is assumed to be linearly related to the time
taken to perform the simulation. Work to empty region i = 1 is: work1 ¼ z
P
l
N P 1
ð
Þ þ 1
2
z
P þ
z
P z
N
l
N
ð7Þ ð7Þ First part of Eq. (7) is emptying from P to 2 in region i = 1 (or N to N -
P + 2), the second part represents emptying sub-domain 1 (or N - P + 1) in
region i = 1. g
For i = 2 region a similar analysis applies but the population size being
worked on by processor 1 has reduced by 1 sub-domain to Fire Technology 2017 Fire Technology 2017 1428 Fire Technology 2017 Fire Technology 2017 Fire Technology 2017 z
P z
N
¼
N
P P
P
z
N ¼
N
P 1
z
N
ð8Þ ð8Þ Similarly the population size worked on by processor 1 for the ith region can be
expressed as Similarly the population size worked on by processor 1 for the ith region can be
expressed as N
P i 1
ð
Þ
z
N
ð9Þ ð9Þ Therefore substituting Z/P in (2) with (9) yields the work done by processor 1
whilst the ith region is emptied. Therefore substituting Z/P in (2) with (9) yields the work done by processor 1
whilst the ith region is emptied. Work to empty region i = 1 is: P 1
2
P
N=P
i¼1
1
"
#
ð16Þ Substituting the series summations Substituting the series summations Substituting the series summations Substituting the series summations speedup ¼
1
2
N2
N2
P 2 1
2
N
P
N
P þ 1
þ N
P
P N
2P
ð17Þ
mplifies to
NP speedup ¼
1
2
N2
N2
P 2 1
2
N
P
N
P þ 1
þ N
P
P N
2P
ð17Þ
mplifies to
speedup ¼
NP
N þ P 1
ð18Þ
E
(18) it
b
th t if N P th
d
ill t
d t
P If N
P speedup ¼
1
2
N2
N2
P 2 1
2
N
P
N
P þ 1
þ N
P
P N
2P
ð17Þ
lifi speedup ¼
1
2
N2
N2
P 2 1
2
N
P
N
P þ 1
þ N
P
P N
2P ð17Þ Simplifies to speedup ¼
NP
N þ P 1
ð18Þ speedup ¼
NP
N þ P 1 ð18Þ From Eq. (18) it can be seen that if N P then speedup will tend to P. If N = P
then then the speedup will tend to P/2. From Eq. (18) it can be seen that if N P then speedup will tend to P. If N = P
then then the speedup will tend to P/2. Work to empty region i = 1 is: So the (computational) speedup over a single processor using P processors over
N sub-domains can be calculated speedup ¼ workserial
total
workN; P
total
ð14Þ speedup ¼ workserial
total
workN; P
total ð14Þ Substituting (12) and (13) into (14) leads to (15) Substituting (12) and (13) into (14) leads to (15) Substituting (12) and (13) into (14) leads to (15) Increasing the Simulation Performance of Large-Scale Evacuations 1429 speedup ¼
1
2
N2
P
N=P
i¼1
N
P i 1
ð
Þ
P 1
ð
Þ þ 1
2
N
P i 1
ð
Þ
þ
N
P i
ð15Þ speedup ¼
1
2
N2
P
N=P
i¼1
N
P i 1
ð
Þ
P 1
ð
Þ þ 1
2
N
P i 1
ð
Þ
þ
N
P i
ð15Þ
speedup ¼
1
2
N2
P
N=P
i¼1
N
P P
N=P
i¼1
i þ P
N=P
i¼1
1
! P 1
2
P
N=P
i¼1
1
"
#
ð16Þ ¼
1
2
N2
P
N=P
i¼1
N
P i 1
ð
Þ
P 1
ð
Þ þ 1
2
N
P i 1
ð
Þ
þ
N
P i
ð15Þ ð15Þ speedup ¼
1
2
N2
P
N=P
i¼1
N
P P
N=P
i¼1
i þ P
N=P
i¼1
1
! P 1
2
P
N=P
i¼1
1
"
#
ð16Þ ð16Þ p ¼
2
N2
P
N=P
i¼1
N
P P
N=P
i¼1
i þ P
N=P
i¼1
1
! General Derivation applicable to Any Number of Sub-domains (N) In the more general case when N is not an integer multiple of P (see Fig. 9) the
analysis is essentially the same although the first region (i = 1) needs to be given
special treatment. The work performed by processor 1 whilst area (a) is emptied is number of peo-
ple being computed by processor 1 9 (distance travelled by an occupant) (19). worka ¼ B
l
N
ð19Þ
ere, worka ¼ B
l
N
ð19Þ where, where, K ¼ N mod P
ð20 K ¼ N mod P
ð20Þ K ¼ N mod P K ¼ N mod P
ð20Þ
M ¼ N K
ð21Þ
H K
ð Þ ¼
0;
if K ¼ 0
1;
if K > 0
ð22Þ K ¼ N mod P ð20Þ ð21Þ M ¼ N K H K
ð Þ ¼
0;
if K ¼ 0
1;
if K > 0
ð22Þ H K
ð Þ ¼
0;
if K ¼ 0
1;
if K > 0
¼
0;
if K ¼ 0
1;
if K > 0
ð22Þ ð22Þ Fire Technology 2017 1430 Fire Technology 2017 Fire Technology 2017 ¼ z
N
M
P þ HðKÞ
ð23Þ B ¼ z
N
M
P þ HðKÞ
ð23Þ In the previous example, i = 1 region there was necessarily P sub-domains. The
number of sub-domains in region 1 is now K. All other regions have P sub-do-
mains. There are [M/P + H(K)] regions. [
/
(
)]
g
The work performed by processor 1 whilst region i = 1 is emptied is: work1 ¼ B l
N P þ K 1 HðKÞP
ð
Þ þ 1
2 B þ
B z
N
l
N
ð24Þ ð24Þ Equation (24) is analogous to (7) with the first part modified due to the number
of sub-domains being reduced to K. P and H(K)P are introduced so the equation
is valid when K = 0. Equation (24) simplifies to work1 ¼ B l
N P þ K HðKÞP
ð
Þ z
2N
l
N
ð25Þ ð25Þ This equation can be generalised for the ith region (27) by substituting the popu-
lation size processed by processor 1 for the ith region (26) into (25) and noting
K = 0 for i > 1. General Derivation applicable to Any Number of Sub-domains (N) This equation can be generalised for the ith region (27) by substituting the popu-
lation size processed by processor 1 for the ith region (26) into (25) and noting
K = 0 for i > 1. B ði 1Þ z
N
ð26Þ B ði 1Þ z
N
ð26Þ worki ¼
B ði 1Þ z
N
l
N P 1
2
z
N
l
N
ð27Þ ð27Þ Total work (28) is the summation of emptying all the regions. The first term in
(28) represents emptying the i = 1 region and the summation term represents
emptying regions 2 to (M/P + H(K)) worktotal ¼
B l
N P þ K HðKÞP
ð
Þ z
2N
l
N
þ
X
M=PþHðKÞ
i¼2
B ði 1Þ z
N
l
N P 1
2
z
N
l
N
ð28Þ ð28Þ This simplifies to (29), note the change in lower summation limit to i = 1. worktotal ¼ B l
N K HðKÞP
ð
Þ þ
X
M=PþHðKÞ
i¼1
B ði 1Þ z
N
l
N P 1
2
z
N
l
N
ð29Þ ð29Þ Increasing the Simulation Performance of Large-Scale Evacuations Increasing the Simulation Performance of Large-Scale Evacuations 1431 g ¼ M
P þ HðKÞ
ð30Þ g ¼ M
P þ HðKÞ ð30Þ Introducing (30) into (29) and simplifying leads to (31). worktotal ¼ zl
N
g
N K HðKÞP
ð
Þ þ
g
N P þ P
N 1
2N
X
g
i¼1
1 P
N
X
g
i¼1
i
! General Derivation applicable to Any Number of Sub-domains (N) Case 1: Idealised Large Geometry
See Tables 3 and 4. Case 1: Idealised Large Geometry
See Tables 3 and 4. Case 2: Long Open Area
See Tables 5, 6, 7 and 8. Table 3
Single Core Timings and Computational Speedup for Case 1
No. computers
Time
taken (m:s)
Max
sub-domains per
processor (MSDPP)
Analytical
computational
Speedup
(=20/MSDPP)
Actual
computational
speedup
Real
time
speedup
1
51:01
20
1
1
0.28
2
21:50
10
2
2.3
0.64
3
14:17
7
2.86
3.6
1.01
4
11:17
5
4
4.5
1.26
5
8:59
4
5
5.7
1.60
7
6:37
3
6.67
7.7
2.2
10
4:40
2
10
10.9
3.1
Table 4
Dual Core Timings and Computational Speedup for Case 1
No. computers
(dual)
Time
taken (m:s)
Max
sub-domains per
PROCESSOR (MSDPP)
Analytical
computational
Speedup
(= 20/MSDPP)
Actual
computational
speedup
Real
time
speedup
1 (1 core)
51:01
20
1
1
0.28
1 (2 cores)
27:32
10
2
1.85
0.52
2 (4 cores)
14:41
5
4
3.47
0.97
3 (6 cores)
11:23
4
5
4.48
1.25
4 (8 cores)
8:58
3
6.67
5.69
1.59
5 (10 cores)
6:39
2
10
7.67
2.15
10 (20 cores)
3:40
1
20
13.9
3.9 Table 3
Single Core Timings and Computational Speedup for Case 1
No. computers
Time
taken (m:s)
Max
sub-domains per
processor (MSDPP)
Analytical
computational
Speedup
(=20/MSDPP)
Actual
computational
speedup
Real
time
speedup
1
51:01
20
1
1
0.28
2
21:50
10
2
2.3
0.64
3
14:17
7
2.86
3.6
1.01
4
11:17
5
4
4.5
1.26
5
8:59
4
5
5.7
1.60
7
6:37
3
6.67
7.7
2.2
10
4:40
2
10
10.9
3.1 Single Core Timings and Computational Speedup for Case 1 Table 4
Dual Core Timings and Computational Speedup for Case 1
No. computers
(dual)
Time
taken (m:s)
Max
sub-domains per
PROCESSOR (MSDPP)
Analytical
computational
Speedup
(= 20/MSDPP)
Actual
computational
speedup
Real
time
speedup
1 (1 core)
51:01
20
1
1
0.28
1 (2 cores)
27:32
10
2
1.85
0.52
2 (4 cores)
14:41
5
4
3.47
0.97
3 (6 cores)
11:23
4
5
4.48
1.25
4 (8 cores)
8:58
3
6.67
5.69
1.59
5 (10 cores)
6:39
2
10
7.67
2.15
10 (20 cores)
3:40
1
20
13.9
3.9 Table 4
Dual Core Timings and Computational Speedup for Case 1 ual Core Timings and Computational Speedup for Case 1 Case 2: Long Open Area
See Tables 5, 6, 7 and 8. General Derivation applicable to Any Number of Sub-domains (N) ð31Þ ð31Þ orktotal ¼ zl
N
g
N K HðKÞP
ð
Þ þ
g
N P þ P
N 1
2N
g P
2N gðg þ 1Þ
ð32Þ ð32Þ Simplifies to Simplifies to worktotal ¼ zl
2N2 g 2 K HðKÞP
ð
Þ þ gP þ P 1
ð
Þ
ð
Þ
ð33Þ ð33Þ Substituting the second g in (33) with (30) Substituting the second g in (33) with (30) Substituting the second g in (33) with (30) worktotal ¼ zl
2N2 g 2K HðKÞP þ M þ P 1
ð
Þ
ð
Þ orktotal ¼ zl
2N2 g 2K HðKÞP þ M þ P 1
ð
Þ
ð
Þ
ð34Þ ð34Þ Substituting g (30) in (34) leads to Substituting g (30) in (34) leads to Substituting g (30) in (34) leads to worktotal ¼ zl
2N2
M
P 2K HðKÞMP
P
þ
M
P M þ MP
P M
P
þHðKÞ2K HðKÞ
ð
Þ2P þ HðKÞM þ HðKÞP HðKÞ
ð35Þ orktotal ¼ zl
2N2
M
P 2K HðKÞMP
P
þ
M
P M þ MP
P M
P
worktotal ¼ 2N2
P 2K
P
þ
P M þ P P
þHðKÞ2K HðKÞ
ð
Þ2P þ HðKÞM þ HðKÞP HðKÞ
ð35Þ ð35Þ þHðKÞ2K HðKÞ
ð
Þ2P þ HðKÞM þ HðKÞP HðKÞ
ð35Þ Simplifying (35) and noting that K HðKÞK and HðKÞ HðKÞ
ð
Þ2 Simplifying (35) and noting that K HðKÞK and HðKÞ HðKÞ
ð
Þ2 workN;P
total ¼
zl
2N2P 2MK þ M M þ P 1
ð
Þ þ 2PK HðKÞP
ð
Þ
ð36Þ workN;P
total ¼
zl
2N2P 2MK þ M M þ P 1
ð
Þ þ 2PK HðKÞP
ð
Þ
ð36Þ ð36Þ uting (36) and (13) into (14) gives the speedup (37) Substituting (36) and (13) into (14) gives the speedup (37) Substituting (36) and (13) into (14) gives the speedup (37) ¼
N 2P
M M þ P 1
ð
Þ þ 2KðM þ PÞ HðKÞP
ð37Þ ð37Þ Substituting (21) into (37) and simplifying leads to (33) Substituting (21) into (37) and simplifying leads to (33) Substituting (21) into (37) and simplifying leads to (33) SN
P ¼
N 2P
N N þ P 1
ð
Þ þ KP K2 HðKÞP
ð38Þ SN
P ¼
N 2P
N N þ P 1
ð
Þ þ KP K2 HðKÞP ð38Þ 1432 Fire Technology 2017 Appendix B: Additional Tables of Results for Case Studies
Case 1: Idealised Large Geometry
See Tables 3 and 4. General Derivation applicable to Any Number of Sub-domains (N) Case 2: Long Open Area
See Tables 5, 6, 7 and 8. Increasing the Simulation Performance of Large-Scale Evacuations 1433 Table 5
Single Core Timings and Computational Speedup for Case 2
Time taken (m:s)
Analytical computa-
tional speedup
Actual computational
speedup
No. partitions fi
20
50
100
20
50
100
20
50
100
No. computers fl
1
11:32
11:32
11:32
1
1
1
1.00
1.00
1.00
2
5:39
5:45
6:18
1.9
1.96
1.98
2.04
2.00
1.83
3
3:53
3:56
4:13
2.76
2.89
2.94
2.97
2.94
2.74
4
2:59
2:55
3:07
3.48
3.78
3.88
3.86
3.96
3.71
5
2:28
2:23
2:30
4.17
4.63
4.8
4.67
4.82
4.60
6
2:13
2:00
2:06
4.78
5.48
5.73
5.18
5.74
5.45
7
1:58
1:43
1:51
5.39
6.26
6.61
5.85
6.66
6.20
8
1:48
1:32
1:34
5.84
7.1
7.5
6.37
7.52
7.33
9
1:37
1:24
1:25
6.37
7.91
8.34
7.15
8.29
8.16
10
1:33
1:19
1:14
6.9
8.47
9.17
7.44
8.74
9.34
Table 6
Single Core Real Time Speedups for Case 2
No computers fi
1
2
3
4
5
6
7
8
9
10
No partitionsfl
20
1.24
2.54
3.69
4.80
5.80
6.44
7.27
7.92
8.89
9.25
50
1.24
2.49
3.65
4.92
5.99
7.13
8.28
9.35
10.30
10.86
100
1.24
2.27
3.41
4.61
5.72
6.77
7.71
9.11
10.14
11.61
Table 7
Dual Core Timings and Computational Speedup for Case 2
Time taken (m:s)
Analytical computa-
tional speedup
Actual computational
speedup
No. partitions fi
20
50
100
20
50
100
20
50
100
No. computers fl
1 (1 core)
11:32
11:32
11:32
1.00
1.00
1.00
1.00
1.00
1.00
1 (2 cores)
6:05
6:19
6:52
1.9
1.96
1.98
1.89
1.83
1.68
2 (4 cores)
3:16
3:14
3:28
3.48
3.78
3.88
3.52
3.56
3.32
3 (6 cores)
2:26
2:13
2:25
4.78
5.48
5.73
4.73
5.19
4.76
4 (8 cores)
2:00
1:43
1:46
5.84
7.1
7.5
5.78
6.72
6.51 Table 5
Single Core Timings and Computational Speedup for Case 2
Time taken (m:s)
Analytical computa-
tional speedup
Actual computational
speedup
No. partitions fi
20
50
100
20
50
100
20
50
100
No. General Derivation applicable to Any Number of Sub-domains (N) Table 8
Dual Core Real Time Speedups for Case 2
No computers fi
1
2
3
4
5
6
7
8
9
10
No partitionsfl
20
2.35
4.37
5.88
7.18
8.60
9.23
9.93
10.2
11.4
11.9
50
2.27
4.42
6.45
8.35
9.53
11.4
12.1
13.3
14.4
14.8
100
2.09
4.13
5.92
8.09
9.86
11.6
13.2
14.9
15.5
17.2
Table 9
Single Core Timings and Speedups for Case 3
Time taken (m:s)
Computational
speedup
Real time speedup
Population size fi
8000
16,000
8000
16,000
8000
16,000
No. computersfl
1
06:57
31:26
1
1
7.21
2.86
2
05:16
18:38
1.32
1.69
9.52
4.84
3
04:05
13:17
1.70
2.37
12.26
6.79
4
03:34
10:40
1.95
2.95
14.06
8.45
5
03:15
09:24
2.14
3.34
15.43
9.56
6
03:07
08:13
2.23
3.83
16.08
10.97
7
02:58
08:03
2.34
3.90
16.87
11.17
8
02:48
07:29
2.48
4.20
17.88
12.03
9
02:48
07:16
2.48
4.33
17.88
12.40
10
02:48
07:00
2.48
4.49
17.88
12.86
Table 10
Dual Core Timings and Speedups for Case 3
Time Taken (m:s)
Computational
speedup
Real time speedup Case 3: High-Rise Building
See Tables 9 and 10. Case 3: High-Rise Building See
a
e 9 a d
0
Table 9
Single Core Timings and Speedups for Case 3
Time taken (m:s)
Computational
speedup
Real time speedup
Population size fi
8000
16,000
8000
16,000
8000
16,000
No. computersfl
1
06:57
31:26
1
1
7.21
2.86
2
05:16
18:38
1.32
1.69
9.52
4.84
3
04:05
13:17
1.70
2.37
12.26
6.79
4
03:34
10:40
1.95
2.95
14.06
8.45
5
03:15
09:24
2.14
3.34
15.43
9.56
6
03:07
08:13
2.23
3.83
16.08
10.97
7
02:58
08:03
2.34
3.90
16.87
11.17
8
02:48
07:29
2.48
4.20
17.88
12.03
9
02:48
07:16
2.48
4.33
17.88
12.40
10
02:48
07:00
2.48
4.49
17.88
12.86
Table 10
Dual Core Timings and Speedups for Case 3
Time Taken (m:s)
Computational
speedup
Real time speedup
Population size fi
8000
16,000
8000
16,000
8000
16,000
No. General Derivation applicable to Any Number of Sub-domains (N) computers fl
1
11:32
11:32
11:32
1
1
1
1.00
1.00
1.00
2
5:39
5:45
6:18
1.9
1.96
1.98
2.04
2.00
1.83
3
3:53
3:56
4:13
2.76
2.89
2.94
2.97
2.94
2.74
4
2:59
2:55
3:07
3.48
3.78
3.88
3.86
3.96
3.71
5
2:28
2:23
2:30
4.17
4.63
4.8
4.67
4.82
4.60
6
2:13
2:00
2:06
4.78
5.48
5.73
5.18
5.74
5.45
7
1:58
1:43
1:51
5.39
6.26
6.61
5.85
6.66
6.20
8
1:48
1:32
1:34
5.84
7.1
7.5
6.37
7.52
7.33
9
1:37
1:24
1:25
6.37
7.91
8.34
7.15
8.29
8.16
10
1:33
1:19
1:14
6.9
8.47
9.17
7.44
8.74
9.34 Table 5
Single Core Timings and Computational Speedup for Case 2 Table 6
Single Core Real Time Speedups for Case 2
No computers fi
1
2
3
4
5
6
7
8
9
10
No partitionsfl
20
1.24
2.54
3.69
4.80
5.80
6.44
7.27
7.92
8.89
9.25
50
1.24
2.49
3.65
4.92
5.99
7.13
8.28
9.35
10.30
10.86
100
1.24
2.27
3.41
4.61
5.72
6.77
7.71
9.11
10.14
11.61 Table 7
Dual Core Timings and Computational Speedup for Case 2
Time taken (m:s)
Analytical computa-
tional speedup
Actual computational
speedup
No. partitions fi
20
50
100
20
50
100
20
50
100
No. computers fl
1 (1 core)
11:32
11:32
11:32
1.00
1.00
1.00
1.00
1.00
1.00
1 (2 cores)
6:05
6:19
6:52
1.9
1.96
1.98
1.89
1.83
1.68
2 (4 cores)
3:16
3:14
3:28
3.48
3.78
3.88
3.52
3.56
3.32
3 (6 cores)
2:26
2:13
2:25
4.78
5.48
5.73
4.73
5.19
4.76
4 (8 cores)
2:00
1:43
1:46
5.84
7.1
7.5
5.78
6.72
6.51
5 (10 cores)
1:40
1:30
1:27
6.9
8.47
9.17
6.92
7.67
7.93
6 (12 cores)
1:33
1:15
1:13
7.5
9.91
10.9
7.43
9.19
9.37
7 (14 cores)
1:26
1:11
1:05
8.07
11.7
12.4
7.99
9.74
10.6
8 (16 cores)
1:24
1:05
0:58
8.74
12.4
14
8.20
10.7
12.0
9 (18 cores)
1:15
1:00
0:55
9.55
15.3
15.7
9.17
11.6
12.5
10 (20 cores)
1:12
0:58
0:50
10.3
15.7
16.8
9.61
11.9
13.8 Table 7
Dual Core Timings and Computational Speedup for Case 2 Dual Core Timings and Computational Speedup for Case 2 Fire Technology 2017 1434 Case 3: High-Rise Building
See Tables 9 and 10. General Derivation applicable to Any Number of Sub-domains (N) National Institute of Standards and Technology (NIST). Technical
Table 12
Timings and Speedups for Case 4 with 120,000 Agents for 10 Single
Core and 1 Dual Core Configuration
No. computers
Time taken (m:s)
Computational speedup
Real time speedup
1
50:13
1
0.43
2
27:40
1.82
0.79
3
20:56
2.40
1.04
4
14:17
3.52
1.53
5
13:34
3.70
1.61
6
13:02
3.85
1.67
7
11:04
4.54
1.97
8
10:19
4.87
2.12
9
14:01
3.58
1.56
10
08:27
5.94
2.58
10 (20 cores)
07:17
6.89
2.96
Table 11
Timings and Speedups for Case 4 with 60,000 Agents
Time taken (m:s)
Computational
speedup
Real time speedup
No. partitions fi
10
20
32
10
20
32
10
20
32
No. computers fl
1
10:25
10:25
10:25
1
1
1
1.0
1.0
1.0
2
06:22
05:44
06:36
1.64
1.82
1.58
1.64
1.81
1.58
3
04:32
04:26
04:45
2.30
2.35
2.19
2.29
2.34
2.18
4
03:55
03:12
04:38
2.66
3.26
2.25
2.65
3.25
2.24
5
03:05
03:24
03:43
3.38
3.06
2.80
3.37
3.05
2.79
6
03:25
03:04
03:23
3.05
3.40
3.08
3.04
3.39
3.07
7
02:38
03:10
02:51
3.96
3.29
3.65
3.95
3.28
3.64
8
02:20
02:44
02:55
4.46
3.81
3.57
4.45
3.80
3.56
9
02:23
02:58
02:39
4.37
3.51
3.93
4.36
3.50
3.92
10
02:18
02:12
02:20
4.53
4.73
4.46
4.52
4.72
4.45
10 (20 cores)
n/a
01:58
02:01
n/a
5.30
5.17
n/a
5.28
5.15 Table 11
Timings and Speedups for Case 4 with 60,000 Agents Table 11
Timings and Speedups for Case 4 with 60,000 Agents
Time taken (m:s)
Computational
speedup
Real time speedup
No. partitions fi
10
20
32
10
20
32
10
20
32
No. General Derivation applicable to Any Number of Sub-domains (N) computersfl
1 (1 core)
06:57
31:26
1
1
7.21
2.86
1 (2 cores)
06:56
22:59
1.00
1.37
7.21
3.92
2 (4 cores)
04:25
13:30
1.57
2.33
11.32
6.66
3 (6 cores)
03:42
10:41
1.88
2.94
13.55
8.41
4 (8 cores)
03:23
09:17
2.05
3.39
14.78
9.70
5 (10 cores)
03:11
08:27
2.18
3.72
15.72
10.64
6(12 cores)
03:03
07:42
2.28
4.08
16.44
11.67
7 (14 cores)
02:56
07:29
2.37
4.20
17.09
12.01
8 (16 cores)
03:00
07:19
2.32
4.30
16.73
12.30
9 (18 cores)
03:07
07:35
2.23
4.15
16.08
11.87
10 (20 cores)
03:10
07:40
2.19
4.10
15.79
11.73 Table 9
Single Core Timings and Speedups for Case 3
Time taken (m:s)
Computational
speedup
Real time speedup
Population size fi
8000
16,000
8000
16,000
8000
16,000
No. computersfl
1
06:57
31:26
1
1
7.21
2.86
2
05:16
18:38
1.32
1.69
9.52
4.84
3
04:05
13:17
1.70
2.37
12.26
6.79
4
03:34
10:40
1.95
2.95
14.06
8.45
5
03:15
09:24
2.14
3.34
15.43
9.56
6
03:07
08:13
2.23
3.83
16.08
10.97
7
02:58
08:03
2.34
3.90
16.87
11.17
8
02:48
07:29
2.48
4.20
17.88
12.03
9
02:48
07:16
2.48
4.33
17.88
12.40
10
02:48
07:00
2.48
4.49
17.88
12.86 Single Core Timings and Speedups for Case 3 Table 10
Dual Core Timings and Speedups for Case 3
Time Taken (m:s)
Computational
speedup
Real time speedup
Population size fi
8000
16,000
8000
16,000
8000
16,000
No. computersfl
1 (1 core)
06:57
31:26
1
1
7.21
2.86
1 (2 cores)
06:56
22:59
1.00
1.37
7.21
3.92
2 (4 cores)
04:25
13:30
1.57
2.33
11.32
6.66
3 (6 cores)
03:42
10:41
1.88
2.94
13.55
8.41
4 (8 cores)
03:23
09:17
2.05
3.39
14.78
9.70
5 (10 cores)
03:11
08:27
2.18
3.72
15.72
10.64
6(12 cores)
03:03
07:42
2.28
4.08
16.44
11.67
7 (14 cores)
02:56
07:29
2.37
4.20
17.09
12.01
8 (16 cores)
03:00
07:19
2.32
4.30
16.73
12.30
9 (18 cores)
03:07
07:35
2.23
4.15
16.08
11.87
10 (20 cores)
03:10
07:40
2.19
4.10
15.79
11.73 1435 Increasing the Simulation Performance of Large-Scale Evacuations Case 4: Large Urban Space
See Tables 11 and 12. References
1. Gwynne S, Galea ER, Owen M, Lawrence PJ, Filippidis L (1999) A review of the
methodologies used in evacuation modeling. Fire Mater. 23(6):383–389. doi:10.1002/
(SICI)1099-1018(199911/12)23:6<383::AID-FAM715>3.0.CO;22
2. Kuligowski ED, Peacock RD, Hoskins B L (2010) A review of building evacuation
models, 2nd edn. General Derivation applicable to Any Number of Sub-domains (N) computers fl
1
10:25
10:25
10:25
1
1
1
1.0
1.0
1.0
2
06:22
05:44
06:36
1.64
1.82
1.58
1.64
1.81
1.58
3
04:32
04:26
04:45
2.30
2.35
2.19
2.29
2.34
2.18
4
03:55
03:12
04:38
2.66
3.26
2.25
2.65
3.25
2.24
5
03:05
03:24
03:43
3.38
3.06
2.80
3.37
3.05
2.79
6
03:25
03:04
03:23
3.05
3.40
3.08
3.04
3.39
3.07
7
02:38
03:10
02:51
3.96
3.29
3.65
3.95
3.28
3.64
8
02:20
02:44
02:55
4.46
3.81
3.57
4.45
3.80
3.56
9
02:23
02:58
02:39
4.37
3.51
3.93
4.36
3.50
3.92
10
02:18
02:12
02:20
4.53
4.73
4.46
4.52
4.72
4.45
10 (20 cores)
n/a
01:58
02:01
n/a
5.30
5.17
n/a
5.28
5.15 able 11
imings and Speedups for Case 4 with 60,000 Agents Table 12
Timings and Speedups for Case 4 with 120,000 Agents for 10 Single
Core and 1 Dual Core Configuration
No. computers
Time taken (m:s)
Computational speedup
Real time speedup
1
50:13
1
0.43
2
27:40
1.82
0.79
3
20:56
2.40
1.04
4
14:17
3.52
1.53
5
13:34
3.70
1.61
6
13:02
3.85
1.67
7
11:04
4.54
1.97
8
10:19
4.87
2.12
9
14:01
3.58
1.56
10
08:27
5.94
2.58
10 (20 cores)
07:17
6.89
2.96 12
gs and Speedups for Case 4 with 120,000 Agents for 10 Single
and 1 Dual Core Configuration 1. Gwynne S, Galea ER, Owen M, Lawrence PJ, Filippidis L (1999) A review of the
methodologies used in evacuation modeling. Fire Mater. 23(6):383–389. doi:10.1002/
(SICI)1099-1018(199911/12)23:6<383::AID-FAM715>3.0.CO;22 2. Kuligowski ED, Peacock RD, Hoskins B L (2010) A review of building evacuation
models, 2nd edn. National Institute of Standards and Technology (NIST). Technical
note 1680, November 2010 1. Gwynne S, Galea ER, Owen M, Lawrence PJ, Filippidis L (1999) A review of the
methodologies used in evacuation modeling. Fire Mater. 23(6):383–389. doi:10.1002/
(SICI)1099-1018(199911/12)23:6<383::AID-FAM715>3.0.CO;22
2. Kuligowski ED, Peacock RD, Hoskins B L (2010) A review of building evacuation
models, 2nd edn. National Institute of Standards and Technology (NIST). Technical
note 1680, November 2010 References 1. Gwynne S, Galea ER, Owen M, Lawrence PJ, Filippidis L (1999) A review of the
methodologies used in evacuation modeling. Fire Mater. 23(6):383–389. doi:10.1002/
(SICI)1099-1018(199911/12)23:6<383::AID-FAM715>3.0.CO;22 2. Kuligowski ED, Peacock RD, Hoskins B L (2010) A review of building evacuation
models, 2nd edn. National Institute of Standards and Technology (NIST). Technical
note 1680, November 2010 1436 Fire Technology 2017 3. Qureshi WS, Ekpanyapong M, Dailey MN, Rinsurongkawong S, Malenichev A, Kra-
sotkina O (2016) QuickBlaze: early fire detection using a combined video processing
approach. Fire Technol 52:1293. doi:10.1007/s10694-015-0489-7 4. Beji T, Verstockt S, Van de Walle R, Merci B (2014) On the use of real-time video to
forecast fire growth in enclosures. fire technol 50:1021. doi:10.1007/s10694-012-0262-0 5. Miller-Hooks E, Krauthammer T (2007) An intelligent evacuation, rescue and recovery
concept. Fire Technol 43:107. doi:10.1007/s10694-006-8433-5 6. Cowlard A, Jahn W, Abecassis-Empis C, Rein G, Torero JL (2010) Sensor assisted fire
fighting. Fire Technol 46:719. doi:10.1007/s10694-008-0069-1 7. Han L, Potter S, Beckett G et al (2010) FireGrid: An e-infrastructure for next-genera-
tion emergency response support. J Parallel Distrib Comput 70(11):1128–1141. doi:
10.1016/j.jpdc.2010.06.005. ISSN 0743-7315 /j jp
8. Galea ER, Xie H, Lawrence P (2016) Intelligent active dynamic signage system: bring-
ing the humble emergency exit sign into the 21st century, SFPE Europe, Q1, Issue 3,
2016. http://www.sfpe.org/general/custom.asp?page=Issue3Feature1 9. Galea ER, Galparsoro JMP (1994) A computer based simulation model for the predic-
tion of evacuation from mass transport vehicles. Fire Saf J 22:341–366. doi:10.1016/
0379-7112(94)90040-X 10. Gwynne S, Galea ER, Lawrence PJ, Filippidis L (2001) Modelling occupant interaction
with fire conditions using the buildingEXODUS evacuation model. Fire Saf J 36:327–
357. doi:10.1016/S0379-7112(00)00060-6 /
(
)
11. Gwynne S, Galea ER, Lawrence PJ (2006) The introduction of social adaptation within
evacuation modelling. Fire Mater 30(4):285–309. doi:10.1002/fam.913 12. Galea ER, Sharp G, Lawrence PJ, Holden R (2008) Approximating the evacuation of
the world trade center north tower using computer simulation. J Fire Prot Eng
18(2):85–115. doi:10.1177/1042391507079343 ( )
/
13. Helbing D, Mukerji P (2012) Crowd disasters as systemic failures: analysis of the Love
Parade disaster. EPJ Data Sci 1(7):1–40. doi:10.1140/epjds7 14. Pretorius M, Gwynne S, Galea E (2012) The collection and analysis of data from a
fatal large-scale crowd incident. In: Proceedings of the 5th international symposium,
human behaviour in Fire 2012, Cambridge UK, 19–21 Sept 2012, Interscience Commu-
nications Ltd., pp 263–274. ISBN: 978-0-9556548-8-6, 2012 15. References Helbing D, Johansson A, Al-Abideen HZ (2007) The dynamics of crowd disasters: an
empirical study. Phys Rev. doi:10.1103/PhysRevE.75.046109 16. Gwynne SMV, Siddiqui AA (2013) Understanding and simulating large crowds. In:
Traffic and granular flow ‘11, 2013, conference proceedings. Springer-Verlag Berlin Hei-
delberg, Berlin Heidelberg, Germany, pp. 217–239. ISBN: 9783642396687 17. Benedictus L (2015) Hajj crush: how crowd disasters happen, and how they can be
avoided. The Guardian, 3 Oct 2015. https://www.theguardian.com/world/2015/oct/03/
hajj-crush-how-crowd-disasters-happen-and-how-they-can-be-avoided. Accessed 3 Jan
2017 18. Lovreglio R, Ronchi E, Maragkos G, Beji T, Merci B (2016) A dynamic approach for
the impact of a toxic gas dispersion hazard considering human behaviour and disper-
sion modelling. J Hazard Mater 318:758–771. doi: 10.1016/j.jhazmat.2016.06.015. ISSN
0304-3894 19. Chooramun N, Lawrence PJ, Galea ER (2012) An agent based evacuation model utilis-
ing hybrid space discretisation. Saf Sci 50:1685–1694. doi:10.1016/j.ssci.2011.12.022 20. Hoogendoorn SP, Bovy PHL (2002) Normative pedestrian behaviour theory and mod-
elling. In: Proceedings of the 15th international symposium on transportation and traf-
fic theory, 2002 1437 Increasing the Simulation Performance of Large-Scale Evacuations 21. Lovreglio R (2016) Modelling decision-making in fire evacuation using the random util-
ity theory. Ph.D. thesis, Politecnico di Bari, Milan and Turin (Italy) 22. Vassalos D, Kim H, Christiansen G, Majumder J (2001) A mesoscopic model for pas-
senger evacuation in a virtual ship-sea environment and performance-based evaluation’,
pedestrian and evacuation dynamics— 4–6 Apr 2001, Duisburg, pp 369–391. ISBN: 3-
540-42690-6 23. Kostreva MM, Lancaster LC (1998) A comparison of two methodologies in HAZARD
I fire egress analysis. Fire Technol 34:227. doi:10.1023/A:1015345923210 24. Kirchner A, Klu¨ pfel H, Nishinari K, Schadschneider A, Schreckenberg M (2002) Simu-
lation of competitive egress behaviour. Phys A 324:689–697. doi:10.1016/S0378-4371
(03)00076-1 (
)
25. Helbing D, Molnar P (1995) Social force model for pedestrian dynamics. Phys Rev E
5:4282–4286. doi:10.1103/PhysRevE.51.4282 26. Chraibi M, Seyfried A, Schadschneider A (2010) Generalized centrifugal-force model
for pedestrian dynamics. Phys Rev E 82(4 pt 2):046111. doi:10.1103/PhysRevE.82. 046111 27. Gao Y, Chen T, Luh PB, Zhang H (2016) Modified social force model based on predic-
tive
collision
avoidance
considering
degree
of
competitiveness. Fire
Technol
. doi:10.1007/s10694-016-0573-7 28. Waterson NP, Mecca A, Wall JM (2004) Evacuation of a multilevel office building:
comparison of predicted results using an agent-based model with measured data. In:
Interflam 2004: 10th international fire science and engineering conference, Edinburgh,
UK, 2004, pp 767–772 29. References Fang ZM, Song WG, Zhang J, Wu H (2012) A multi-grid model for evacuation coupling
with the effects of fire products. Fire Technol 48:91. doi:10.1007/s10694-010-0173-x 30. Zia K, Farrahi K, Riener A, Ferscha A (2013) An agent-based parallel geo-simulation
of urban mobility during city-scale evacuation. Simulation: transactions of the society
for
modeling
and
simulation
international,
May
2013,
pp
1–31. doi:10.1177/
0037549713485468 31. Duncan R (1990) A survey of parallel computer architectures. Computer 23(2):5–16. doi:10.1109/2.44900 32. Giitsidis T, Dourvas NI, Sirakoulis GC (2015) Parallel implementation of aircraft dis-
embarking and emergency evacuation based on cellular automata. Int J High Perform
Comput Appl. doi:10.1177/1094342015584533 33. Chen D, Wang L, Zomaya AY, Dou M, Chen J, Deng Z, Hariri S (2015) Parallel simu-
lation of complex evacuation scenarios with adaptive agent models. IEEE Trans Paral-
lel Distrib Syst 26(3):847–857. doi:10.1109/TPDS.2014.2311805 34. Quinn MJ, Metoyer RA, Hunter-Zaworski K (2003) Parallel implementation of the
social forces model. In Galea ER (ed) Proceedings of 2nd international pedestrian and
evacuation dynamics conference. CMS Press, Greenwich, UK, pp 63–74. ISBN:
1904521088 35. Steffen B, Kemloh U, Chraibi M, Seyfried A (2011) Parallel real time computation of
large scale pedestrian evacuations. In: Iva´ nyi P, Topping (BHV) (eds) Proceedings of
the second international conference on parallel, distributed, grid and cloud computing
for engineering, Civil-Comp Press, Stirlingshire, UK, paper 95. doi:10.4203/ccp.95.95 36. Dagum L, Menon R (1998) OpenMP: an industry standard API for shared-memory
programming. IEEE Computat Sci Eng 5(1):46–55. doi:10.1109/99.660313 37. Galea ER, Ierotheou C (1992) Fire field modelling on parallel computers. Fire Saf J
19(4):251–266. doi:10.1016/0379-7112(92)90008-Z 1438 Fire Technology 2017 38. Grandison AJ, Galea ER, Patel MK, Ewer J (2003–2004) The development of parallel
implementation for a CFD based fire field model utilising conventional office based
PCs. J Appl Fire Sci 12(2):137–157. doi:10.2190/AGH5-EXUR-J110-HPHE 39. Grandison AJ, Galea ER, Patel MK, Ewer J (2007) Parallel CFD fire modelling on
office PCs with dynamic load balancing. Int J Numer Meth Fluids 55:29–39. doi:10.1002/fld.1278 40. McGrattan K, Hostikka S, McDermott R, Floyd J, Weinschenk C, Overholt K
(XXXX) Fire Dynamics simulator user’s guide. NIST special publication 1019, 6th edn. doi: 10.6028/NIST.SP.1019 41. Message Passing Interface Forum (2015) MPI: a message-passing interface standard,
version 3.1. High Performance Computing Centre Stuttgart, 2015, EAN: 11144444100
30 42. Gropp W, Lusk E, Doss N, Skjellum A (1996) A high-performance, portable implemen-
tation of the MPI message passing interface standard. Parallel Comput 22:789–828. doi:10.1016/0167-8191(96)00024-5 43. References Amdahl GM (1967) Validity of the single-processor approach to achieving large scale
computing
capabilities. In:
Proceedings
of
AFIPS,
pp
483–485. doi:10.1145/
1465482.1465560 44. Grandison A, Muthu Y, Lawrence P, Galea E (2007) Simulating the evacuation of very
large populations in large domains using a parallel implementation of the build-
ingEXODUS evacuation model. In: Proceedings of the 11th international fire science &
engineering conference, Interflam 2007, 3–5th Sept 2007, Royal Holloway College,
University of London, UK, vol 1, pp 259–270. ISBN: 978 0 9541216-8-6 45. Nagashima U, Hyugaji S, Sekiguchi S, Sato M, Hosoya H (1995) An experience with
super-linear speedup achieved by parallel computing on a workstation cluster: parallel
calculation of density of states of large scale cyclic polyacenes. Parallel Comput
21(9):1491–1504. doi:10.1016/0167-8191(95)00026-K 46. Helmbold DP, McDowell CE (1989) Modeling speedup(n) greater than n. 1989 interna-
tional conference on parallel processing proceedings, vol III, pp 219–225 47. Johnson SP (1992) Mapping numerical software onto distributed memory parallel sys-
tems. Ph.D. thesis, University of Greenwich 48. Karypis G, Kumar V (1998) Multilevel k-way partitioning scheme for irregular graphs. J Parallel Distrib Comput 48(1):96–129. doi:10.1006/jpdc.1997.1404 48. Karypis G, Kumar V (1998) Multilevel k-way partitioning scheme for irregular graphs. J Parallel Distrib Comput 48(1):96–129. doi:10.1006/jpdc.1997.1404
|
https://openalex.org/W1963957951
|
https://www.scielo.br/j/rbcs/a/nFfbZLtG8GTxSQ5Q6QkC3XG/?lang=pt&format=pdf
|
Portuguese
| null |
Adsorção e dessorção de cádmio, cobre e chumbo por amostras de Latossolos pré-tratadas com fósforo
|
Revista Brasileira de Ciência do Solo
| 2,004
|
cc-by
| 6,433
|
M. A. P. PIERANGELI(2), L. R. G. GUILHERME(3), N. CURI(3),
S. J. ANDERSON(4) & J. M. LIMA(3) M. A. P. PIERANGELI(2), L. R. G. GUILHERME(3), N. CURI(3),
S. J. ANDERSON(4) & J. M. LIMA(3) Termos de indexação: ligante inorgânico, metais pesados, imobilização de cádmo,
cobre e chumbo, sorção, poluição do solo. (1) Trabalho apresentado no 7th ICOBT (7th International Conference on the Biogeochemistry of Trace Elements, Uppsala, Sweden,
15-19 June, 2003. Recebido para publicação em outubro de 2002 e aprovado em janeiro de 2004.
(2) Professor Visitante da Universidade Estadual do Mato Grosso, UNEMAT, Caixa Postal 92, CEP 78250-000 Pontes e Lacerda
(MT). E-mail: mapp2@terra.com.br
(3) Professor do Departamento de Ciência do Solo, Universidade Federal de Lavras – UFLA. Caixa Postal 37, CEP 37200-000
Lavras (MG). Bolsista do CNPq. E-mails: guilherm@ufla.br; niltcuri@ufla.br; jmlima@ufla.br
(4) Earth Systems Sci. and Policy, CSUMB, 100 Campus Ctr., Seaside, CA 93955-8001. E-mail: sharon_anderson@csumb.cdu fessor do Departamento de Ciência do Solo, Universidade Federal de Lavras – UFLA. Caixa Postal 37,
vras (MG). Bolsista do CNPq. E-mails: guilherm@ufla.br; niltcuri@ufla.br; jmlima@ufla.br Systems Sci. and Policy, CSUMB, 100 Campus Ctr., Seaside, CA 93955-8001. E-mail: sharon_anderson@cs i. and Policy, CSUMB, 100 Campus Ctr., Seaside, CA 93955-8001. E-mail: sharon_anderson@csumb.cdu te da Universidade Estadual do Mato Grosso, UNEMAT, Caixa Postal 92, CEP 78250-000 Pontes e Lacerd
app2@terra.com.br R. Bras. Ci. Solo, 28:377-384, 2004 (1) Trabalho apresentado no 7th ICOBT (7th International Conference on the Biogeochemistry of Trace Elements, Uppsala, Sweden,
15-19 June, 2003. Recebido para publicação em outubro de 2002 e aprovado em janeiro de 2004. amento de Ciência do Solo, Universidade Federal de Lavras – UFLA. Caixa Postal 37, CEP 37200-000
ta do CNPq. E-mails: guilherm@ufla.br; niltcuri@ufla.br; jmlima@ufla.br INTRODUÇÃO A adsorção de fosfatos em sistemas com
predomínio de cargas positivas, por exemplo,
horizonte B de alguns Latossolos, aumenta a carga
líquida negativa e promove a elevação da adsorção
de Cu (Guilherme & Anderson, 1998). Já, em sistemas
com carga líquida negativa, por exemplo, horizonte
A de Latossolos, o efeito da adsorção de fosfatos sobre
a carga líquida é menor (Lima et al., 2000). Elementos-traço ocorrem naturalmente nos solos
e alguns deles, tais como: Cu, Zn e Co, desempenham
importante papel na nutrição de plantas e animais,
enquanto outros, como Cd, Pb, As e Se, exercem
efeitos deletérios sobre vários componentes da
biosfera (Alloway, 1990; Kabata-Pendias & Pendias,
2001). Na maioria das vezes, estes elementos estão
presentes nos solos em concentrações que não
oferecem risco para o ambiente. Todavia, nas últimas
décadas, atividades antropogênicas têm elevado,
substancialmente, a concentração de alguns
elementos-traço em diversos ecossistemas (Kabata-
Pendias & Pendias, 2001). Um ânion, quando adicionado ao solo antes do
metal (M = Cd, Cu, Pb, etc), como, por exemplo, por
meio de fertilização fosfatada antes da aplicação de
fungicida cúprico, poderá ligar-se aos óxidos (S) de
Fe ou Al presentes, alterar a carga líquida
superficial de positiva para negativa (Chorover,
1997) e promover a formação de complexos ternários
do tipo S-fosfato-Mcomplexos (Kuo, 1986; McBride,
1994; Guilherme & Anderson, 1998). Efeitos
benéficos de fosfato na redução de injúrias
provocadas por fungicidas cúpricos são relatados na
literatura (Chaney & Giordano, 1977). Tradicionalmente, o enriquecimento do solo com
estes elementos se relaciona com as atividades
industriais e mineradoras (Alloway, 1990). No entanto,
vários trabalhos (Amaral Sobrinho et al., 1992; Lima,
2001) têm destacado a presença de alguns destes
elementos, dentre eles Cd, Cu e Pb, em alguns
materiais usados para melhoria das condições
químicas e físicas dos solos. Assim, a aplicação de
calcário, lodo de esgoto, fertilizantes fosfatados,
resíduos industriais, pesticidas e aterros sanitários
são exemplos de práticas que podem, eventualmente,
elevar a concentração de elementos-traço nos solos
(Alloway, 1990; Kabata-Pendias & Pendias, 2001). Nos solos, a mobilidade destes elementos depende,
fundamentalmente, das reações de adsorção e
dessorção que ocorrem entre eles e os componentes
sólidos do sistema. Estas reações são influenciadas
por vários fatores, merecendo destaque a presença de
ligantes orgânicos e inorgânicos (Alloway, 1990;
McBride, 1994). A influência de substâncias inorgânicas,
especialmente fosfatos, na adsorção e dessorção de
Cd em solos é reportada por Krishnamurti et al. RESUMO O movimento de elementos-traço nos solos está intimamente associado aos
fatores que contribuem para sua retenção na fase sólida; dentre esses fatores,
destaca-se a presença de ânions adsorvidos. Neste trabalho, avaliou-se o efeito
do pré-tratamento do solo com P sobre a adsorção e dessorção de Cd, Cu e Pb em
amostras dos horizontes A e B de dois solos: Latossolo Vermelho-Amarelo ácrico
típico (LVAw) e Latossolo Vermelho ácrico típico (LVw). O pré-tratamento com
P consistiu na reação das amostras de solo, durante 72 h, com Ca(H2PO4)2.H2O
10,75 mmol L-1 (relação solo:solução de 1:1,5). Para a adsorção dos metais,
amostras de solo tratadas e não tratadas com P foram inicialmente pré-
equilibradas com Ca(NO3)2 5 mmol L-1 (relação solo:solução 1:67, pH 5,5) e
colocadas para reagir, individualmente, com Cu(NO3)2, Cd(NO3)2 ou Pb(NO3)2 na
concentração de 0,45 mmol L-1 (relação solo:solução final 1:100). O pré-tratamento
com P aumentou a adsorção de todos os metais, com maior incremento nas
amostras dos horizontes B. O aumento da adsorção com o tratamento com P
indica que a presença de P nesses solos contribui para reduzir a disponibilidade
dos metais Cd, Cu e Pb. A quantidade adsorvida segue a seqüência Pb ≥≥≥≥≥ Cu
>> Cd. Com relação à fração de metal dessorvida (metaldessorvido/metaladsorvido),
esta foi menor nas amostras previamente tratadas com P para Pb, sendo a
dessorção de Cu não influenciada pelo tratamento com P. Termos de indexação: ligante inorgânico, metais pesados, imobilização de cádmo,
cobre e chumbo, sorção, poluição do solo. R. Bras. Ci. Solo, 28:377-384, 2004 378 M.A.P. PIERANGELI et al. ABSTRACT: CADMIUM, COPPER, AND LEAD ADSORPTION AND
DESORPTION IN OXISOL SAMPLES PRE-TREATED WITH
PHOPHORUS The movement of trace elements in soils is closely related to factors that contribute to
their retention in the solid phase, for instance adsorbed anions. In the present work, it was
evaluated the effect of phosphorus (P) pre-treatment on adsorption and desorption of Cd,
Cu, and Pb in A and B horizon samples of two Latosol (Oxisols): a Yellow-Red Latosol and
a Dark-Red Latosol. The P pre-treatment consisted of reacting soils samples for 72 h with
10.75 mmol L-1 Ca(H2PO4)2.H2O (soil:solution ratio of 1:1.5). For adsorbing the metals, samples
of pre-treated and non pre-treated soil material were pre-equilibrated with 5 mmol L-1
Ca(NO3)2 (soil:solution ratio of 1:67; pH 5.5), and then reacted, individually, with
0.450 mmol L-1 Cu(NO3)2, Cd(NO3)2, or Pb(NO3)2, (final soil:solution ratio of 1:100). Phosphorus pre-treatment increased the adsorption of all metals, particularly in B-horizon
samples. The increased adsorption indicates that the presence of P in these soils may
contribute to reduce Cd, Cu and Pb availability. The amount of metal adsorbed followed
the sequence Pb ≥ Cu >> Cd. Regarding the fraction of desorbed metal (desorbed metal/
adsorbed metal), it was smaller in the previously P treated samples, except for Cu, whose
desorption was not influenced by the pre-treatment with P. Index terms: inorganic ligant, P, Cd, Cu, and Pb immobilization, sorption, soil pollution. Index terms: inorganic ligant, P, Cd, Cu, and Pb immobilization, sorption, soil pollution. MATERIAL E MÉTODOS Amostras dos horizontes A e B de um Latossolo
Vermelho-Amarelo ácrico típico (LVAw) e de um
Latossolo Vermelho ácrico típico (LVw) (EMBRAPA,
1999) foram coletadas próximo a São João Del Rei,
na zona fisiográfica Campos das Vertentes, em
Minas Gerais, Brasil (latitude 21 º 20 ’ S; longitude
44 º 30 ’ W). Estes solos, originados de rochas
metapelíticas pobres, apresentam teores de argila e
de óxidos de Fe semelhantes (Quadro 1). Informações sobre os métodos usados para a
caracterização química, física e mineralógica destes
solos podem ser encontradas em Guilherme &
Anderson (1998) e Lima & Anderson (1997). Além de aumentar a adsorção e, conseqüente-
mente, diminuir a mobilidade e biodisponibilidade
de Cd, Cu e Pb em solos, resíduos e outros sistemas,
o P é, principalmente, usado para induzir a formação
de compostos insolúveis desses metais (Ma et al.,
1993; Crannell et al., 2000). De acordo com Nriagu
(1984), fosfatos de Pb e Zn têm solubilidade de várias
ordens de magnitude menor que análogos carbonatos
e sulfatos. Dentro desta premissa, o uso de fosfatos
como aditivos para remediação, in situ, de solos
contaminados com Cd, Cu e Pb tem sido muito
pesquisado nos últimos anos (Ma et al., 1993; Xu et
al., 1994; Cotter-Howells, 1996; Laperche et al., 1996;
Boisson et al., 1999). No entanto, para a formação
de fosfatos de metal, é preciso que o P esteja presente
como íon fosfato livre, HPO42-, na solução do solo
(Cotter-Howells & Caporn, 1996). Segundo esses
autores, até mesmo o fosfato que está disponível para
as plantas poderia, por meio de vários mecanismos
(Häeussling & Marschner, 1989; Jayachandran et
al., 1992), combinar com alguns elementos-traço
formando fosfatos de metais. Um dos aspectos
interessantes da formação de fosfatos de metais para
imobilização desses elementos é que, segundo
Nriagu (1984), os fosfatos de metais são estáveis com
relação ao pH, Eh e diagênese mineral. Amostras de cada solo foram colocadas para
reagir com Ca(H2PO4)2.H2O na concentração de
10,75 mmol L-1 em uma relação solo:solução de 1:1,5
(m:v) (adição de 1 g kg-1 de P no solo). As amostras
permaneceram reagindo por 72 h, alternando-se
12 h de agitação e 12 h de repouso. Após esse tempo,
elas foram centrifugadas durante 30 min a
3.000 rpm (1.609,72 g) para sedimentar os colóides
maiores que 0,45 µm. ADSORÇÃO E DESSORÇÃO DE CÁDMIO, COBRE E CHUMBO POR AMOSTRAS... 379 relataram diminuição na quantidade trocável de Zn,
Pb, Cu e Cd, em amostras de solos contaminados,
tratados com hidroxiapatita. Segundo Cotter-
Howells & Caporn (1996) e Ma (1996), o uso de
fosfatos em solos contaminados com Pb pode reduzir
a biodisponibilidade deste elemento, graças à
formação de fosfato de Pb (por exemplo, piromorfita). resíduos contaminados nos solos, quando não houver
outras alternativas econômicas e ambientalmente
mais viáveis. Tendo em vista a importância do conhecimento
dos fatores que atuam na retenção e, ou, imobilização
de Cd, Cu e Pb em solos, o objetivo deste trabalho
foi avaliar o efeito do pré-tratamento com P sobre a
adsorção e dessorção desses elementos em amostras
dos horizontes A e B de dois Latossolos com teores
de argila e óxidos de Fe semelhantes. A elevação da adsorção de Zn, Cd e Cu por óxidos
de Fe e de Al, baixa ou moderadamente enriquecidos
com fosfato, tem sido verificada (Bolland et al., 1977;
Kuo & McNeal., 1984; McBride, 1985). Entretanto,
a extrapolação desses resultados obtidos com
sorventes puros para sistemas naturais heterogêneos
é duvidosa. Tal incerteza decorre da íntima
associação da matéria orgânica com os grupos
funcionais dos óxidos metálicos nos solos. Além
disso, de acordo com Guilherme & Anderson (1998),
os óxidos puros sintéticos não são bons modelos para
óxidos pedogênicos. INTRODUÇÃO (1999), os quais constataram que a fertilização
fosfatada poderia aumentar a disponibilidade deste
elemento nos solos. Em solos que receberam lodo
de esgoto, Gonzalez et al. (1992) verificaram redução
da biodisponibilidade de Cd na presença de argila
rica em fosfato. Trabalhando com cinzas que
continham altos teores de elementos-traço,
provenientes de resíduos urbanos, Crannell et al. (2000) verificaram, após tratamento do resíduo com
fosfato, redução da fração de metal disponível para
lixiviação em cerca de 14 % para Cd, 98 % para Cu
e 99 % para Pb. Também, Boisson et al. (1999) R. Bras. Ci. Solo, 28:377-384, 2004 MATERIAL E MÉTODOS O excesso de solução foi
removido da amostra e a concentração de P no
sobrenadante foi medida colorimetricamente (Ce1),
conforme EMBRAPA (1997). Os frascos, com o
resíduo remanescente após a centrifugação, foram
pesados para determinar a massa da solução
residual. Posteriormente, estes resíduos foram lavados com
Ca(NO3)2 5 mmol L-1 (relação solo:solução 1:2, m:v)
para extração do P prontamente dessorvido, cuja
concentração também foi determinada colorimetrica-
mente (Ce2), conforme foi supracitado. A quantidade
de P adsorvida (média de 30 mmol kg-1 de P no solo
para cada amostra) foi calculada pela expressão:
Padsorvido = [Padicionado - (Ce1/V1 + Ce2/V2)/kg solo], em
que V1 (L) é o volume da solução adicionada para a
realização da adsorção de P e V2 (L) é a solução
adicionada para a obtenção do P prontamente
dessorvido mais o restante de solução que ficou
retida junto à massa de solo, após a retirada do
sobrenadante para determinação do P adsorvido. As
pastas de solo, obtidas após a lavagem com Ca(NO3)2
5 mmol L-1, foram secas ao ar, tendo sido os torrões O efeito da pré-adsorção de fosfatos na retenção
dos metais Pb e Cd, em solos tropicais, com alta
capacidade de retenção de P é desconhecido. No
entanto, como pode ser visto, a utilização do P na
redução da mobilidade de metais em áreas já
contaminadas é amplamente relatada na literatura,
tendo o P a capacidade de amenizar os efeitos tóxicos
de Cu, Pb e Cd, dentre outros elementos, tanto por
aumentar a adsorção, quanto por possibilitar a
formação de compostos insolúveis. Sendo assim, a
presença de P em áreas sujeitas a receber
quantidades elevadas destes elementos poderia ser
útil para minimizar a lixiviação e biodisponibilidade
destes elementos, viabilizando, assim, o descarte de R. Bras. Ci. Solo, 28:377-384, 2004 M.A.P. PIERANGELI et al. 380 Quadro 1. MATERIAL E MÉTODOS Atributos físicos, químicos e mineralógicos e eletroquímicos de amostras dos Latossolos
estudados
PESN
Solo
Argila
CO(2)
FeDCB
Feox
Feox/FeDCB
Ct
Gb
Gt:Hm
ASE
∆pH
Sem P
Com P
__________________ g kg-1 ___________________
_ g kg-1 argila _
m2 g-1
LVAwA(1)
711
15,8
101
1,20
0,0108
480
375
10,2
54 ± 3
-
4,3
3,8
LVAwB
721
9,2
114
0,59
0,0052
480
400
8,3
54 ± 2
1,1
6,2
5,3
LVwA
691
18,8
99
1,88
0,0190
350
510
5,0
64 ± 5
-
4,3
4,0
LVwB
753
11,0
114
0,65
0,0057
350
510
4,1
63 ± 2
0,65
6,1
5,8
Fonte: Adaptado de Guilherme & Anderson (1998); Lima & Anderson (1997). (1) LVAw = Latossolo Vermelho-Amarelo ácrico típico; LVw = Latossolo Vermelho ácrico típico; os índices A e B indicam os horizon-
tes. (2) CO = Carbono orgânico; FeDCB e Feox = Ferro extraído pelo ditionito-citrato-bicarbonato de sódio e oxalato ácido de amônio,
respectivamente; Ct = caulinita; Gb = gibbsita; Gt:Hm = relação goethita:hematita; ASE = área superficial específica.; ∆pH = pHKCl
– pHH2O; PESN = ponto de efeito salino nulo. Quadro 1. Atributos físicos, químicos e mineralógicos e eletroquímicos de amostras dos Latossolos
estudados cos, químicos e mineralógicos e eletroquímicos de amostras dos Latossolos p
(1) LVAw = Latossolo Vermelho-Amarelo ácrico típico; LVw = Latossolo Vermelho ácrico típico; os índices A e B indicam os horizon-
tes. (2) CO = Carbono orgânico; FeDCB e Feox = Ferro extraído pelo ditionito-citrato-bicarbonato de sódio e oxalato ácido de amônio,
respectivamente; Ct = caulinita; Gb = gibbsita; Gt:Hm = relação goethita:hematita; ASE = área superficial específica.; ∆pH = pHKCl
– pHH2O; PESN = ponto de efeito salino nulo. triturados e peneirados em malha de 2 mm. Esse
material constituiu as amostras pré-tratadas com
P. No decorrer do experimento, foram mantidas
amostras-controle, nas quais o pH era medido
diariamente. Elevações do pH indicariam alterações
nas condições redox do meio, o que poderia interferir
nos resultados. Para a adsorção de Cd, Cu ou Pb, foram colocadas
0,3 g das amostras de solos não tratadas e pré-
tratadas com P (conforme anteriormente descrito),
em triplicata, em suspensão com Ca(NO3)2 5 mmol L-1
(relação solo:solução 1:67, m:v) e tiveram o pH
ajustado para 5,5 (± 0,2) com solução de Ca(OH)2
11,12 mmol L-1. MATERIAL E MÉTODOS Em seguida, foram adicionadas,
individualmente em cada amostra, soluções de
Cu(NO3)2, Cd(NO3)2 ou Pb(NO3)2 nas concentrações
de 0,45 mmol L-1, visando obter uma concentração
inicial de cada metal equivalente a 0,15 mmol L-1
(relação solo:solução final 1:100, m:v). Após 72 h de
reação, alternando-se 12 h de agitação e 12 h de
repouso, as amostras foram centrifugadas em
temperatura ambiente a 1.609,72 g durante 20 min
e o sobrenadante coletado para determinação do teor
dos metais por espectrofotometria de absorção
atômica de chama. A quantidade de metal adsorvida
foi calculada pela diferença entre a quantidade de
metal adicionada e a quantidade remanescente na
solução de equilíbrio. O delineamento experimental foi em esquema
fatorial (4 x 2 x 3) inteiramente casualizado, sendo:
4 amostras de solos (dois solos, horizontes A e B),
duas situações (pré-tratadas e não tratadas com P)
e 3 metais (Cd, Cu ou Pb). Estatisticamente,
analisou-se o efeito do tratamento com P e dos solos
e respectivos horizontes sobre as quantidades
adsorvidas e dessorvidas de Cd, Cu ou Pb. R. Bras. Ci. Solo, 28:377-384, 2004 ADSORÇÃO E DESSORÇÃO DE CÁDMIO, COBRE E CHUMBO POR AMOSTRAS... (LVAw = Latossolo Vermelho-Amarelo ácrico típico; LVw = Latossolo Vermelho ácrico
típico; os índices A e B indicam os horizontes e P as amostras tratadas com fósforo). Colunas seguidas
da mesma letra, para um mesmo metal, não diferem entre si pelo teste de Scott-Knott a 5 %. Barras
acima das colunas indicam o erro-padrão da média (n = 3). Figura 1. Quantidade adsorvida dos metais Cd, Cu e Pb por amostras de Latossolos, tratadas e não tratadas
com fósforo, suspensas em Ca(NO3)2 5 mmol L-1 (pH 5,5, relação solo:solução 1:67), após colocadas
para reagir, individualmente, com Cd(NO3)2, Cu(NO3)2 e Pb(NO3)2 0,45 mmol L-1 (relação solo:solução
final 1:100). (LVAw = Latossolo Vermelho-Amarelo ácrico típico; LVw = Latossolo Vermelho ácrico
típico; os índices A e B indicam os horizontes e P as amostras tratadas com fósforo). Colunas seguidas
da mesma letra, para um mesmo metal, não diferem entre si pelo teste de Scott-Knott a 5 %. Barras
acima das colunas indicam o erro-padrão da média (n = 3). Maiores quantidades adsorvidas dos três metais,
observadas nas amostras do horizonte A, de ambos
os solos, exceto Pb no LVAw, cuja quantidade
adsorvida não diferiu entre os horizontes,
provavelmente, estão relacionadas com a maior
carga líquida negativa presente no horizonte A,
quando comparadas às do horizonte B, haja vista o
balanço de carga positivo apresentado por estes
(Quadro 1). O ponto de efeito salino nulo (PESN)
do horizonte A dos dois solos, menor que no
horizonte B (4,3 vs 6,2), também constitui uma
evidência da maior quantidade de cargas negativas
nestes. Contribui para isso, sem dúvida, a maior
quantidade de carbono orgânico presente neste
horizonte [cerca de 1,7 vez mais que o
correspondente horizonte B (Quadro 1)]. As
correlações observadas entre a quantidade de metal
adsorvida e carbono orgânico e PESN corroboram
esta afirmação (Quadro 2). A menor quantidade de
metal adsorvida nas amostras dos horizontes B, por
outro lado, provavelmente, está relacionada com a
maior quantidade de cargas positivas destes (Lima
& Anderson, 1997; Lima et al., 2000). O aumento
da quantidade adsorvida observada nas amostras
tratadas com P, o qual ocupa os sítios com carga
positiva, viabiliza esta possibilidade. RESULTADOS E DISCUSSÃO Observou-se que a adsorção, tanto nos solos
tratados com P quanto nos não tratados, segue a
seqüência Pb ≥ Cu >> Cd (Figura 1). Vale ressaltar
que a comparação ora realizada só é pertinente,
porque foram utilizadas, nos experimentos, iguais
concentrações molares dos três metais, os valores
estão expressos em mmol kg-1 e, na reação em
questão, Cu e Pb estão na forma divalente (Cd tem
somente valência 2). Tal fato não seria possível se
se optasse pela notação em mg do metal adsorvido
por kg de solo, uma vez que a massa atômica difere
significativamente entre eles. Ressalta-se que o Cd
foi muito pouco adsorvido nas amostras do horizonte
B não tratadas com P. O Pb foi ligeiramente mais
adsorvido que o Cu, notadamente nas amostras não
tratadas com P. Para os mesmos horizontes e mesma
condição de solo (isto é, tratados ou não tratados com
P), tanto a adsorção de Cd quanto a de Cu e Pb não
diferiram estatisticamente entre os solos, exceto
para Pb, nas amostras de horizontes B não tratadas
com P. Nessas, o LVwB adsorveu menor quantidade
de Pb que o LVAwB. Ao resíduo remanescente do experimento de
adsorção com os metais, o qual foi pesado para
determinação do volume de solução e da massa de
solo retida, foram adicionados 30 mL de solução de
Ca(NO3)2 5 mmol L-1 (pH ajustado para 5,5 com
solução de Ca(OH)2 11,12 mmol L-1) para a dessorção
dos metais nas amostras de solo, repetindo-se o
mesmo procedimento com relação ao período de
agitação e repouso realizado durante a adsorção. As
amostras foram, então, centrifugadas, conforme já
descrito, e o sobrenadante foi coletado para leitura
da concentração dos metais. As quantidades de Cd,
Cu e Pb dessorvidas foram calculadas, descontado-
se a quantidade de cada metal retida na solução,
após a retirada do sobrenadante no experimento de
adsorção dos metais. R. Bras. Ci. Solo, 28:377-384, 2004 ADSORÇÃO E DESSORÇÃO DE CÁDMIO, COBRE E CHUMBO POR AMOSTRAS... ADSORÇÃO E DESSORÇÃO DE CÁDMIO, COBRE E CHUMBO POR AMOSTRAS... 381 Pb
Cu
Cd
0
2
4
6
8
10
12
14
16
a
b
a
b
a
b
a
b
a
a
a
b
a
a
a
b
a
a
a
b
a
a
a
b
LVAwB+P
LVAwA+P
LVAwB
LVAwA
LVwA+P
LVwA
LVwB+P
LVwB
LVAwB+P
LVAwA+P
LVAwB
LVAwA
LVwA+P
LVwA
LVwB+P
LVwB
LVAwB+P
LVAwA+P
LVAwB
LVAwA
LVwA+P
LVwA
LVwB+P
LVwB
METAL ADSORVIDO, mmol kg-1
Figura 1. Quantidade adsorvida dos metais Cd, Cu e Pb por amostras de Latossolos, tratadas e não tratadas
com fósforo, suspensas em Ca(NO3)2 5 mmol L-1 (pH 5,5, relação solo:solução 1:67), após colocadas
para reagir, individualmente, com Cd(NO3)2, Cu(NO3)2 e Pb(NO3)2 0,45 mmol L-1 (relação solo:solução
final 1:100). (LVAw = Latossolo Vermelho-Amarelo ácrico típico; LVw = Latossolo Vermelho ácrico
típico; os índices A e B indicam os horizontes e P as amostras tratadas com fósforo). Colunas seguidas
da mesma letra, para um mesmo metal, não diferem entre si pelo teste de Scott-Knott a 5 %. Barras
acima das colunas indicam o erro-padrão da média (n = 3). SOLO Pb
Cu
Cd
0
2
4
6
8
10
12
14
16
a
b
a
b
a
b
a
b
a
a
a
b
a
a
a
b
a
a
a
b
a
a
a
b
LVAwB+P
LVAwA+P
LVAwB
LVAwA
LVwA+P
LVwA
LVwB+P
LVwB
LVAwB+P
LVAwA+P
LVAwB
LVAwA
LVwA+P
LVwA
LVwB+P
LVwB
LVAwB+P
LVAwA+P
LVAwB
LVAwA
LVwA+P
LVwA
LVwB+P
LVwB
METAL ADSORVIDO, mmol kg-1
SOLO Figura 1. Quantidade adsorvida dos metais Cd, Cu e Pb por amostras de Latossolos, tratadas e não tratadas
com fósforo, suspensas em Ca(NO3)2 5 mmol L-1 (pH 5,5, relação solo:solução 1:67), após colocadas
para reagir, individualmente, com Cd(NO3)2, Cu(NO3)2 e Pb(NO3)2 0,45 mmol L-1 (relação solo:solução
final 1:100). (LVAw = Latossolo Vermelho-Amarelo ácrico típico; LVw = Latossolo Vermelho ácrico
típico; os índices A e B indicam os horizontes e P as amostras tratadas com fósforo). Colunas seguidas
da mesma letra, para um mesmo metal, não diferem entre si pelo teste de Scott-Knott a 5 %. Barras
acima das colunas indicam o erro-padrão da média (n = 3). Figura 1. Quantidade adsorvida dos metais Cd, Cu e Pb por amostras de Latossolos, tratadas e não tratadas
com fósforo, suspensas em Ca(NO3)2 5 mmol L-1 (pH 5,5, relação solo:solução 1:67), após colocadas
para reagir, individualmente, com Cd(NO3)2, Cu(NO3)2 e Pb(NO3)2 0,45 mmol L-1 (relação solo:solução
final 1:100). ADSORÇÃO E DESSORÇÃO DE CÁDMIO, COBRE E CHUMBO POR AMOSTRAS... Na literatura,
é relatado que o P é capaz de neutralizar parcial ou
totalmente ou, até mesmo, reverter a carga positiva
predominante na superfície de óxidos de Fe e de Al,
em condições de solos ácidos (Wann & Uehara, 1978;
Chorover et al., 1997; Lima et al., 2000). 1,1 e 1,2 vezes nas amostras do horizonte A e de 31,0;
2,0 e 1,9 vezes nas amostras do horizonte B para
Cd, Cu e Pb, respectivamente. Observaram-se os
maiores incrementos no horizonte B, notadamente
para o metal Cd. Com relação a este metal, no
entanto, mesmo nas amostras tratadas com P, a
percentagem adsorvida foi sempre muito baixa
(cerca de 17 %). Krishnamurti et al. (1999)
verificaram redução na adsorção de Cd com a
presença de P. Todavia, a comparação dos resultados
é dificultada por causa das diferentes condições
experimentais, uma vez que no referido trabalho as
soluções que continham Cd e P foram adicionadas
simultaneamente às amostras de solo. As maiores quantidades adsorvidas verificadas
para o Cu e Pb, tanto nas amostras tratadas quanto
nas não tratadas com P, estão de acordo com os dados
da literatura os quais reportam alta afinidade dos
solos para com Cu (Abd-Elfattah & Wada, 1981;
Guilherme & Anderson, 1998) e Pb (Abd-Elfattah &
Wada, 1981; Matos et al., 1996; Pierangeli et al.,
2001a). Isso pode estar relacionado com o fato de
serem os metais Pb (Abd-Elfattah & Wada, 1981;
Pierangeli et al., 2001b) e Cu (Abd-Elfattah & Wada,
1981; Matos et al., 1996; Guilherme & Anderson,
1998) adsorvidos de forma específica na maioria dos
solos. Maior tendência para hidrolizar e maior
eletronegatividade de Cu e Pb ajudam a explicar a
maior afinidade destes para com os solos (McBride,
1994) em relação ao Cd. Altos teores de Cd na
solução do solo em áreas agrícolas, mesmo na
presença de alto P e N, indicando baixa afinidade
de alguns solos para com o Cd, são relatados na O pré-tratamento dos solos com P promoveu
incrementos médios na quantidade adsorvida de 2,4; R. Bras. Ci. Solo, 28:377-384, 2004 382 M.A.P. PIERANGELI et al. Quadro 2. ADSORÇÃO E DESSORÇÃO DE CÁDMIO, COBRE E CHUMBO POR AMOSTRAS... Coeficientes de correlação de Pearson entre a quantidade de metal adsorvida por dois Latossolos
ácricos, tratados e não tratados com fósforo, e alguns de seus atributos químicos, físicos e
mineralógicos
Amostra não tratada com P
Amostra pré-tratada com P
Atributo(1)
Cd
Cu
Pb
Cd
Cu
Pb
Argila
-0,80
-0,89
-0,89
-0,66
-0,70
-0,66
Carbono orgânico
0,92*
0,92*
0,90*
0,88
0,99**
0,94*
FeDCB
-0,98**
-0,98**
-0,97*
-0,95*
-0,97*
-0,96*
Feox
0,86
0,89
0,87
0,78
0,93*
0,85
Feox/FeDCB
0,82
0,86
0,84
0,73
0,91*
0,82
Caulinita
0,05
0,07
0,11
0,04
-0,29
-0,13
Gibbsita
-0,15
-0,15
-0,19
-0,16
0,18
0,01
Gt:Hm
0,33
0,33
0,37
0,34
-0,00
0,17
Área sup. específica
0,02
-0,01
-0,05
-0,07
0,33
0,17
PESN
-0,99**
-0,97*
-0,97*
-0,94*
-0,86
-0,90*
(1) FeDCB e Feox = Ferro extraído pelo ditionito-citrato-bicarbonato de sódio e oxalato ácido de amônio, respectivamente; Gt:Hm =
relação goethita:hematita; PESN = ponto de efeito salino nulo. **;* significativos a 1 e 5 %, respectivamente. Quadro 2. Coeficientes de correlação de Pearson entre a quantidade de metal adsorvida por dois Latossolos
ácricos, tratados e não tratados com fósforo, e alguns de seus atributos químicos, físicos e
mineralógicos Quadro 2. Coeficientes de correlação de Pearson entre a quantidade de metal adsorvida por dois Latossolos
ácricos, tratados e não tratados com fósforo, e alguns de seus atributos químicos, físicos e
mineralógicos Quadro 2. Coeficientes de correlação de Pearson entre a quantidade de metal adsorvida por dois Latossolos
ácricos, tratados e não tratados com fósforo, e alguns de seus atributos químicos, físicos e
mineralógicos (1) FeDCB e Feox = Ferro extraído pelo ditionito-citrato-bicarbonato de sódio e oxalato ácido de amônio, respectivamente; Gt:Hm =
relação goethita:hematita; PESN = ponto de efeito salino nulo. **;* significativos a 1 e 5 %, respectivamente. A fração de Cu e Pb dessorvida (Metaldessorvido/
Metaladsorvido) (Figura 2) foi sempre muito pequena
(em média, < 10 e < 20 %, para Cu e Pb,
respectivamente), sendo o Cu dessorvido de maneira
semelhante em todas as amostras de solos, tratadas
e não tratadas com P. O Pb, no entanto, apresentou
uma fração dessorvida significativamente menor (em
média, 3,5 vezes) nas amostras tratadas com P. Baixas quantidades dessorvidas de Cu e Pb podem
indicar a formação de complexos de esfera interna,
os quais requerem alta energia de ativação para a
dessorção (McBride, 1989). A quantidade dessorvida
de Cd ficou abaixo do nível de detecção do aparelho
de absorção atômica de chama. ADSORÇÃO E DESSORÇÃO DE CÁDMIO, COBRE E CHUMBO POR AMOSTRAS... literatura (Taylor & Percival, 2001). No entanto,
Dias et al. (2001) verificaram forte interação
específica (interpretação com base na isoterma do tipo
H-High apresentada pelo mesmo) entre as partículas
de um Nitossolo Vermelho eutroférrico e o Cd. Prováveis mecanismos para explicar o aumento
da retenção e, ou, imobilização dos metais, na
presença de P, incluem: (a) formação de fosfatos de
metais (Nriagu, 1984; Crannell et al., 2000); (b)
aumento da carga líquida negativa (McBride, 1994)
e (c) disponibilização de grupos fosfatos de superfície,
nos quais os metais podem-se ligar formando
complexos ternários (Kuo, 1986; McBride, 1994;
Guilherme & Anderson, 1998). O fato de a quantidade de metal adsorvido ser
sempre maior nas amostras tratadas com P indica
que a adição de fosfatos, em áreas que estão sujeitas
a receber resíduos que contenham Cd, Cu e Pb, pode
ser efetiva na prevenção da lixiviação e
biodisponibilização destes elementos, ainda que os
mecanismos responsáveis pelo aumento da retenção
não sejam totalmente entendidos. Considerando que o aumento da carga líquida
negativa tenha sido um dos fatores responsáveis pelo
aumento da adsorção de Cd, Cu e Pb, o qual pode
ser inferido pela diminuição do PESN nas amostras
tratadas com P (Quadro 1), o menor incremento na
adsorção, observado nas amostras do horizonte A,
pode ser explicado com base em Lima et al. (1997),
para os quais, o efeito da adsorção de fosfatos sobre
a carga líquida é menor no horizonte A. O aumento
na adsorção de Cd em amostras de goethita pré-
tratada com P foi atribuído, por Wang & Xing (2002),
à redução do ponto de carga zero (≈ PESN)
promovido pelo tratamento. CONCLUSÕES Fração dessorvida (Metaldessorvido/Metaladsorvido), por Ca(NO3)2 5 mmol L-1, dos metais Pb e Cu,
de amostras de Latossolos, tratadas e não tratadas com fósforo, submetidas previamente à adsorção,
individualmente, de Cd(NO3)2, Cu(NO3)2 e Pb(NO3)2 0,45 mmol L-1 (LVAw = Latossolo Vermelho-
Amarelo ácrico típico; LVw = Latossolo Vermelho ácrico típico; os índices A e B indicam os horizontes
e P as amostras tratadas com fósforo). Colunas seguidas da mesma letra, para um mesmo solo, não
diferem entre si pelo teste de Scott-Knott a 5 %. Barras acima das colunas indicam o erro-padrão da
média (n = 3) e a linha pontilhada representa a fração dessorvida média. Figura 2. Fração dessorvida (Metaldessorvido/Metaladsorvido), por Ca(NO3)2 5 mmol L-1, dos metais Pb e Cu,
de amostras de Latossolos, tratadas e não tratadas com fósforo, submetidas previamente à adsorção,
individualmente, de Cd(NO3)2, Cu(NO3)2 e Pb(NO3)2 0,45 mmol L-1 (LVAw = Latossolo Vermelho-
Amarelo ácrico típico; LVw = Latossolo Vermelho ácrico típico; os índices A e B indicam os horizontes
e P as amostras tratadas com fósforo). Colunas seguidas da mesma letra, para um mesmo solo, não
diferem entre si pelo teste de Scott-Knott a 5 %. Barras acima das colunas indicam o erro-padrão da
média (n = 3) e a linha pontilhada representa a fração dessorvida média. COTTER-HOWELLS, J. Lead phosphate formation in soils. Environ. Poll., 93:9-16, 1996. CONCLUSÕES 1. O pré-tratamento com P proporcionou aumento
na adsorção de Cd, Cu e Pb, sendo o incremento
muito maior nas amostras do horizonte B dos
Latossolos estudados. No entanto, conforme Skopp (1986), é difícil e
complicado, com base em simples experimentos de
adsorção, tais como os deste trabalho, fazer
considerações a respeito dos mecanismos envolvidos
nas reações de equilíbrio. Além das propriedades
adsorventes dos múltiplos componentes dos solos, as
interações entre eles, bem como o efeito competitivo
dos vários íons presentes, também precisam ser
consideradas. 2. A quantidade adsorvida seguiu a seqüência
Pb ≥ Cu >> Cd. 2. A quantidade adsorvida seguiu a seqüência
Pb ≥ Cu >> Cd. 3. O pré-tratamento com P reduziu a fração de
Pb dessorvida, o mesmo não se verificando para o
Cu. R. Bras. Ci. Solo, 28:377-384, 2004 ADSORÇÃO E DESSORÇÃO DE CÁDMIO, COBRE E CHUMBO POR AMOSTRAS... 383 1
0,0
0,1
0,2
0,3
0,4
Sem P
Com P
Pb
Cu
LVAwB
LVAwA
LVwA
LVwB
LVAwB
LVAwA
LVwA
LVwB
a
a
a
a
a
a
a a
a
a
b
a
b
b
b
b
b
b
b
SOLO
FRAÇÃO DE METAL DESSORVIDO,
Metaldessorvido/Metaladssorvido Figura 2. Fração dessorvida (Metaldessorvido/Metaladsorvido), por Ca(NO3)2 5 mmol L-1, dos metais Pb e Cu,
de amostras de Latossolos, tratadas e não tratadas com fósforo, submetidas previamente à adsorção,
individualmente, de Cd(NO3)2, Cu(NO3)2 e Pb(NO3)2 0,45 mmol L-1 (LVAw = Latossolo Vermelho-
Amarelo ácrico típico; LVw = Latossolo Vermelho ácrico típico; os índices A e B indicam os horizontes
e P as amostras tratadas com fósforo). Colunas seguidas da mesma letra, para um mesmo solo, não
diferem entre si pelo teste de Scott-Knott a 5 %. Barras acima das colunas indicam o erro-padrão da
média (n = 3) e a linha pontilhada representa a fração dessorvida média. 0,0
0,1
0,2
0,3
0,4
Sem P
Com P
Pb
Cu
LVAwB
LVAwA
LVwA
LVwB
LVAwB
LVAwA
LVwA
LVwB
a
a
a
a
a
a
a a
a
a
b
a
b
b
b
b
b
b
b
SOLO
FRAÇÃO DE METAL DESSORVIDO,
Metaldessorvido/Metaladssorvido Figura 2. LITERATURA CITADA COTTER-HOWELLS, J. & CAPORN, S. Remediation of
contaminated land by formation of heavy metal phosphates. Appl. Geochem., 11:335-342, 1996. ABD-ELFATTAH, A. & WADA, K. Adsorption of lead, copper,
zinc, cobalt, and cadmium by soils that differ in cation-
exchange materials. J. Soil Sci., 32:271-283, 1981. CRANNELL, S.B.; EIGHMY, T.T.; KRZANOWSKI, J.E.;
EUSDEN Jr., D.J.; SHAW, E.L. & FRANCIS, C.A. Heavy
metal stabilization in municipal solid waste combustion
bottom ash using soluble phosphate. Waste Maneg., 20:135-
148, 2000. ALLOWAY, B.J. Heavy metals in soils. In: ALLOWAY, B.J.,
ed. New York, John Wiley & Sons, 1990. 390p. AMARAL SOBRINHO, N.M.B.; COSTA, L.M.; OLIVEIRA, C. &
VELLOSO, A.C.X. Metais pesados em alguns fertilizantes
e corretivos. R. Bras. Ci. Solo, 16:271-276, 1992. DIAS, N.M.P.; ALLEONI, L.R.F.; CASAGRANDE, J.C. &
CAMARGO, O.A. Adsorção de cádmio em dois Latossolos
ácricos e um Nitossolo. R. Bras. Ci. Solo, 25:297-304, 2001. BOISSON, J.; RUTTENS, A.; MENCH, M. & VANGRONSVELD,
J. Evaluation of hydroxyapatite as a metal immobilizing
soil additive for the remediation of polluted soils. Part 1. Influence of hydroxyapatite on metal exchangeability in
soil, plant growth and plant metal accumulation. Environ. Poll., 104:225-233, 1999. EMPRESA DE PESQUISA AGROPECUÁRIA BRASILEIRA -
EMBRAPA. Manual de métodos de análise de solo. Rio de
Janeiro, Centro Nacional de Pesquisa de Solos, 1997. 212p. EMPRESA DE PESQUISA AGROPECUÁRIA BRASILEIRA -
EMBRAPA. Sistema brasileiro de classificação de solos. Brasília, Serviço de Produção de Informação, 1999.412p. BOLLAND, M.D.A.; POSNER, A.M. & QUIRK, J.P. Zinc
adsorption by goethite in the absence and presence of
phosphate. Aust. J. Soil Res., 15:279-286, 1977. GONZALEZ, R.X.; SERTAIN, J.B. & MILLER, W.L. Cadmium
availability and extractability from sewage sludge as
affected by waste phosphatic clay. J. Environ. Quality,
21:272-275, 1992. CHANEY, R.L. & GIORDANO, P.M. Microelements as related
to plant deficiencies and toxicities. In: ELLIOT, L.F. &
STEVENSION, F.J., eds. Soils for management of organic
wastes and wastes waters. Madison, American Society of
Agronomy, 1977. p.234-279. GUILHERME, L.R.G. & ANDERSON, S.J. Copper sorption
kinetics and sorption hysteresis in two oxide-rich soils
(Oxisols): Effect of phosphate pretreatment. In: JENNE,
E.A., ed. Adsorption of metals by geomedia: variables,
mechanisms, and model applications. San Diego, Academic
Press, 1998. p.209-228. CHOROVER, J.; ZHANG, J.; AMISTADI, M.K. & BUFFLE, J. Comparison of hematite coagulation by charge screening
and phosphate adsorption: Differences in aggregate
structure. Clays Clay Miner., 45:690-708, 1997. R. Bras. Ci. Solo, 28:377-384, 2004 M.A.P. PIERANGELI et al. LITERATURA CITADA 384 MATOS, A.T.; FONTES, M.P.F.; JORDÃO, C.P. & COSTA, L.M. Mobilidade e formas de retenção de metais pesados em
Latossolo Vermelho-Amarelo. R. Bras. Ci. Solo, 20:379-386,
1996. HÄEUSSLING M. & MARSCHNER, H. Organic and inorganic
soil phosphates and acid phosphatase activity in the
rhizosphere of a 80-year-old Norway spruce [Picea abies
(L.) Karst.] trees. Biol. Fertil. Soils, 8:128-133, 1989. JAYACHANDRAN, K.; SCHWAB, A.P. & HETRICK, D.A.D. Mineralization of organic phosphorus by vesicular-
arbuscular mycorrhizal fungi. Soil Biol. Biochem, 24:897-
903, 1992. McBRIDE, M.B. Sorption of copper (II) on aluminum hydroxide
as affected by phosphate. Soil Sci. Soc. Am. J., 49:843-846,
1985. McBRIDE, M.B. Reactions controlling heavy metals in soils. Adv. Soil Sci., 10:1-56, 1989. KABATA-PENDIAS, A. & PENDIAS, H. Trace elements in soils
and plants. 3. ed. Boca Raton, CRC Press, 2001. 413p. McBRIDE, M.B. Environmental chemistry of soils. New York,
Oxford University Press, 1994. 406p. KUO, S. Concurrent sorption of phosphate and zinc, cadmium
or calcium by a hydrous ferric oxide. Soil Sci. Soc. Am. J.,
50:1412-1419, 1986. NRIAGU, J.O. Formation and stability of base metal phosphates
in soil and sediments. In: NRIAGU, J.O. & MOORE, P.B.,
eds. Phosphates minerals. London, Springer, 1984. p.318-
329. KUO, S. & McNEAL, B.L. Effects of pH and phosphate on
cadmium sorption by a hydrous ferric oxide. Soil Sci. Soc. Am. J., 48:1040-1044, 1984. PIERANGELI, M.A.P.; GUILHERME, L.R.G.; CURI, N.; SILVA,
M.L.N.; OLIVEIRA, L.R. & LIMA, J.M. Teor total e
capacidade máxima de adsorção de chumbo em Latossolos
brasileiros. R. Bras. Ci. Solo, 25:279-288, 2001a. KRISHNAMURTI, G.S.; HUANG, P.M. & KOZAK, L.M. Sorption and desorption kinetics of cadmium from soils:
influence of phosphate. Soil Sci., 164:888-898, 1999. LAPERCHE, V.; TRAINA, S.J.; GADDAM, P. & LOGAN, T.J. Chemical and mineralogical characterizations of Pb in a
contaminated soil: reactions with synthetic apatite. Environ. Sci. Technol., 30:3321-3326, 1996. PIERANGELI, M.A.P.; GUILHERME, L.R.G.; OLIVEIRA, L.R.;
CURI, N. & SILVA, M.L.N. Efeito da força iônica da solução
de equilíbrio sobre a adsorção/dessorção de chumbo em
Latossolos brasileiros. Pesq. Agropec. Bras., 36:1077-1084,
2001b. LIMA, G.J.E.O. Biodisponibilidade de cádmio, zinco e chumbo
presentes em calcário comercial para o feijoeiro cultivado
em três solos do Noroeste de Minas Gerais. Lavras, Univer-
sidade Federal de Lavras, 2001. 148p. (Tese de Mestrado) SKOPP, J. Analysis of time-dependence process in soils. J. Environ. Quality, 15:205-213, 1986. TAYLOR, M.O. & PERCIVAL, H.J. R. Bras. Ci. Solo, 28:377-384, 2004 LITERATURA CITADA Cadmium in soil solutions
from a transect of soils away from a fertilizer bin. Environ. Poll., 113:35-40, 2001. LIMA, J.M. & ANDERSON, S.J. Effect of aggregation and
aggregate size on extractable Fe and Al in two Brazilian
Typic Hapludoxs. Soil Sci. Soc. Am. J., 61:965-970, 1997. WANG, K. & XING, B. Adsorption and desorption of cadmium
by goethite pretreated with phosphate. Chemosphere,
48:665-670, 2002. LIMA, J.M.; ANDERSON, S.J. & CURI, N. Phosphate-induced
clay dispersion as related to aggregate size and composition
in Hapludoxs. Soil Sci. Soc. Am. J., 64:892-897, 2000. WANN, S.S. & UEHARA, G. Surface charge manipulation of
constant surface potential soil colloids. I: Relation to sorbed
phosphorus. Soil Sci. Soc. Am. J., 42:565-570, 1978. MA, Q.Y.; TRAINA, S.J. & LOGAN, T.J. In situ lead
immobilization by apatite. Environ. Sci. Technol., 27:1803-
1810, 1993. MA, L.Q. Factors influencing the effectiveness and stability of
aqueous lead immobilization by hidroxyapatite. J. Environ. Qual., 25:1420-1429, 1996. XU, Y.; SCHWARTZ, F.W. & TRAINA, S.J. Sorption of Zn2+
and Cd2+ on hydroxyapatite surfaces. Environ. Sci. Tecnol.,
28:1472-1480, 1994.
|
https://openalex.org/W2159805158
|
http://irep.ntu.ac.uk/id/eprint/31703/1/PubSub9156_Hennes.pdf
|
English
| null |
The physiological effects of hypobaric hypoxia versus normobaric hypoxia: a systematic review of crossover trials
|
Extreme physiology & medicine
| 2,015
|
cc-by
| 11,984
|
REVIEW Open Access The physiological effects of hypobaric hypoxia
versus normobaric hypoxia: a systematic review
of crossover trials Jonny Coppel1*, Philip Hennis1, Edward Gilbert-Kawai1 and Michael PW Grocott1,2,3,4 © 2015 Coppel et al.; licensee BioMed Central. This is an Open Access article distributed under the terms of the Creative
Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain
Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article,
unless otherwise stated. Abstract Much hypoxia research has been carried out at high altitude in a hypobaric hypoxia (HH) environment. Many
research teams seek to replicate high-altitude conditions at lower altitudes in either hypobaric hypoxic conditions
or normobaric hypoxic (NH) laboratories. Implicit in this approach is the assumption that the only relevant condition
that differs between these settings is the partial pressure of oxygen (PO2), which is commonly presumed to be the
principal physiological stimulus to adaptation at high altitude. This systematic review is the first to present an overview
of the current available literature regarding crossover studies relating to the different effects of HH and NH on human
physiology. After applying our inclusion and exclusion criteria, 13 studies were deemed eligible for inclusion. Several
studies reported a number of variables (e.g. minute ventilation and NO levels) that were different between the two
conditions, lending support to the notion that true physiological difference is indeed present. However, the presence
of confounding factors such as time spent in hypoxia, temperature, and humidity, and the limited statistical power due
to small sample sizes, limit the conclusions that can be drawn from these findings. Standardisation of the study
methods and reporting may aid interpretation of future studies and thereby improve the quality of data in this area. This is important to improve the quality of data that is used for improving the understanding of hypoxia
tolerance, both at altitude and in the clinical setting. Keywords: Normobaric hypoxia, Hypobaric hypoxia, Altitude conditions, the hypoxic dose is calculated by the com-
bination of the various barometric pressures × inspired
fraction of oxygen [4]. As emphasised in Conkin's ‘Cri-
tique of the equivalent air altitude model’ [5], implicit in
this approach is the assumption that the only relevant
condition that differs between these settings is the par-
tial pressure of oxygen (PO2), which is commonly pre-
sumed to be the principal physiological stimulus to
adaptation at high altitude [6]. Although this assumption
underpins the interpretation of many studies that form
the basis of hypoxia physiology, it remains open to ques-
tion as recently highlighted by Millet et al. [7] and con-
troversy exists relating to the sporadic data in this area
with various opinions on the matter as discussed in a re-
cent series of ‘point-counterpoints’ [8]. The notion that
HH and NH environments are interchangeable in terms
of their effect on physiological responses is not proven. Coppel et al. Extreme Physiology & Medicine (2015) 4:2
DOI 10.1186/s13728-014-0021-6 Coppel et al. Extreme Physiology & Medicine (2015) 4:2
DOI 10.1186/s13728-014-0021-6 * Correspondence: jonny.coppel@gmail.com
1University College London Centre for Altitude Space and Extreme
Environment Medicine, UCLH NIHR Biomedical Research Centre, Institute of
Sport and Exercise Health, 170 Tottenham Court Road, London W1T 7HA, UK
Full list of author information is available at the end of the article Types of outcome measures Outcome variables were any human physiological re-
sponse to atmospheric hypoxia. These responses included
common phenotypes of interest in high altitude literature
including (but not limited to) ventilation, hypoxaemia, ex-
ercise metabolism, nitric oxide (NO) production, osmotic
balance, erythropoiesis and high-altitude illness. The aim of this systematic review is to conduct a com-
prehensive systematic literature search to address the
questions: do humans react differently to HH when
compared with NH (when evaluated in studies with a
crossover design)? Data collection and analysis
Selection of studies We included studies involving lowland (defined as per-
manently living in locations <2,000 m) human subjects
of any age who were not acclimatised to high altitude. Titles and abstracts of candidate studies were screened
for eligibility and duplicate references independently by
two authors (JC and EG). The reasons for study exclu-
sion were independently documented. For those papers
that could not be excluded based on their titles and ab-
stracts, the full paper was read to confirm eligibility. We resolved disagreements by consulting a third author Search terms We searched for primary research articles describing
crossover trials comparing physiological responses to
NH and HH. Only crossover trials were considered due
to the large inter-subject variation in their response to
hypoxia. Search terms include (‘Hypobari* hypoxia’ OR ‘simulated
altitude’ OR ‘hypobari* anoxia’) AND (‘normbari* hyp-
oxia’ OR ‘sea level hypoxia’ OR ‘sea-level hypoxia’ OR
‘normobari* anoxia’). Background Hypoxia research has numerous applications. These in-
clude investigating the pathogenesis and developing treat-
ments for medical conditions characterised by hypoxia [1]
and acute high altitude illness [2], as well as setting
optimum training regimes for athletes [3]. Much hypoxia research has been carried out at high
altitude in a hypobaric hypoxia (HH) environment. Such
‘field’ studies present practical and logistical challenges
including safety concerns about carrying out invasive
procedures in a remote setting. For these reasons, many
research teams seek to replicate high-altitude conditions
at lower altitudes in either hypobaric hypoxic conditions
or normobaric hypoxic (NH) laboratories. In these two Coppel et al. Extreme Physiology & Medicine (2015) 4:2 Coppel et al. Extreme Physiology & Medicine (2015) 4:2 Page 2 of 20 HH may have been performed either at sea level in a
hypobaric chamber or at high altitude. The practical outcomes of this debate affect a variety of
fields. Many national teams in various sports incorporate
altitude or hypoxic training into their programmes to aid
haematological adaptations [3]. Additionally, armies across
the world employ pre-acclimatisation strategies to train
troops for deployment at high altitude [9]. This issue also
applies to medical research such as therapeutic intermit-
tent hypoxic methods [10] or critical care research into
tissue hypoxia [1]. Thus, understanding the different im-
pacts of NH and HH on physiology is important. Criteria for considering studies for this review Criteria for considering studies for this review Candidate studies were identified using the following cri-
teria (Figure 1). Search methods for identification of studies
Search strategy We attempted to identify all relevant trials regardless of
language or publication status (published, unpublished,
in press, and in progress). A literature search was car-
ried out using the search engines Embase (all to date),
Medline (performed on 15 October 2013) and Web of
Science (performed on 15 October 2013). Snowballing
was carried out; thus, the reference lists of all the short-
listed studies were checked for possible eligible studies. Data extraction and management Using data extraction forms, JC extracted information
from each study and EG crosschecked the data. Data
fields within the data extraction forms were directly
linked to the formulated review question and planned
assessment of included studies. The studies
investigated
simulated
altitudes
from
1,700 m to 7,620 m, and exposure to the hypoxia lasted
between 5 min to 24 h. The 13 studies were carried out
in seven different countries: Australia (1), Japan (1),
Spain (1), Sweden (1), Switzerland (2), France (2), and
America (5). The data extraction forms contained the following infor-
mation: study reference and reviewer identity, verification
of study eligibility, study characteristics, study quality (see
Quality of data section below), research methods, partici-
pants,
intervention,
outcome
measures,
results,
and
additional information. Data synthesis
h
l The results were tabulated and compared. No statistical
analysis was carried out because the heterogeneity of the
studies was such that the data could not be pooled (see
below). Studies were categorised according to duration
of hypoxic exposure. One hour was used as the cut off
between ‘long’ and ‘short’ studies. All variables identified
through our search strategy are highlighted in Table 1
(List of variables) but due to the number of variables re-
ported, not all are considered in the written results and
discussion. Emphasis is placed on the major physio-
logical variables and those that are reported in more
than one study. This was because a consistent result
across multiple independent studies suggests validity of
the finding. Additionally, when study characteristics
were being determined, if a study did not mention a de-
sign feature, such as randomization of exposure order, it
was assumed that the feature was not present. The re-
sults for each variable and time point can be found in
Tables 2, 3, 4, 5, 6. The methods used to control the degree of hypoxia
administered in each study varied. Five studies men-
tioned controlling the relative humidity between HH
and NH. Of these, three [15,17,18] maintained 50% hu-
midity (±1%) and two [13,14] maintained it between 45%
and 55%. One paper [6] specifically mentioned the meas-
urement and control of CO2 levels in the chambers
using CO2 scrubbers. Review: results
Studies A total of 225 unique articles were identified in the
EMBASE, MEDLINE and Web of Science searches. After applying our inclusion and exclusion criteria, 13
studies were deemed eligible for inclusion (Table 2:
Study design). A total of 153 subjects were included in
our review. Of these, six subjects were women and nine
had chronic airflow limitations. One study was added
after snowballing [12]. Variables measured
Quality of data
S
d
d
i
Ni The risk of bias was independently assessed by JC and EG. Studies that do not report statistical significance (P values)
for reported results were included, but their results were
considered either as high risk of bias or unclear. In terms
of study quality, we assessed the following: randomisation
of subjects for the order of the crossover and whether they
were blinded, length of washout period, presence/absence
of sample size calculations, whether the statistical analyses
accounted for the increased risk of type I errors when ana-
lysing large numbers of variables (adjustment for multiple
comparisons), test-retest reliability, normalisation of test-
ing environments for humidity (pH2O) as this can impact
hypoxic dose calculations [11], and control of carbon
dioxide (CO2) in the testing environments. Study design Nine studies [12-20] randomised the order
of the crossover. Three studies [6,21,22] did not, and one
[23] was ambiguous as to whether randomization was
used or not. Three studies [12,18,22] had a washout
period of at least 14 days, six studies [13-17,21] used 7
days, and four studies [6,19,20,23] used less than 24 h. The largest study involved 20 people, the smallest 7,
and the mean was 12. None of the studies stated they
had conducted a sample size calculation to justify their
chosen number. Only two studies [12,23] mentioned ac-
counting for the inflated risk of type I errors that arises
when multiple comparisons are made, and both of these
performed Bonferroni adjustments. Only one study mea-
sured the test-retest reliability of their outcome variable
[23]. They performed an intra-class coefficient correl-
ation and showed a good reliability of the postural stabil-
ity measurements. Types of interventions We compared NH and HH. NH and HH must be calcu-
lated to be equivalent to the same altitude. We included
studies investigating any duration of exposure, and the Figure 1 Methodology flow chart. This flow chart shows search methodology and results. igure 1 Methodology flow chart. This flow chart shows search methodology and results. Page 3 of 20 Page 3 of 20 Coppel et al. Extreme Physiology & Medicine (2015) 4:2 (PH) who arbitrated on inclusion. We obtained the as-
sistance of translators when abstracts were not available
in English. Ventilation Eight studies were identified that reported ventilation
and oxygenation. Five of these lasted ≥1 h (long studies)
[12,13,15,22,16], and three lasted <1 h (short studies)
[17,18,21]. Five out of seven studies reporting minute
ventilation reported values that were significantly lower
in HH [12,13,17,21,22] (by up to 4 L/min) [13] (Figure 2:
Graph of minute ventilation), whereas two identified no
difference between conditions [15,18]. Consistent with Page 4 of 20 Coppel et al. Extreme Physiology & Medicine (2015) 4:2 this, the tidal volume was lower in HH in five out of six
volume found in a study was 0.9 L [12
Table 1 List of variables
Oxygenation and ventilation
Cardiovascular
AMS (acute mountain sickness)
and neurology
Other
Ventilation (VE)
Hypoxic cardiac response (HCR)
Postural stability
Sweat rates
Tidal volume (VT)
Forced vascular co
Lake Louise AMS scores
Oesophageal tem
for increasing fore
conductance
Respiratory rate (Bf)
Heart rate variability (LH/HF)
Exhaled nitric oxide (NO) levels
(exNO)
Oesophageal tem
for increased swe
Heart rate (HR)
Oesophageal tem
End tidal partial pressure of oxygen
(PetO2)
Stroke volume (SV)
Skin temperature
Urine volume
Cardiac output (CO)
Plasma volume
End tidal partial pressure of carbon
dioxide (PETCO2)
Glomerular filtratio
Plasma potassium
Plasma sodium co
Blood pressure (BP)
Plasma renin activ
Plasma aldosteron
Alveolar ventilation (VA)
Free water clearan
Adrenocorticotrop
Volume of CO2 produced (VCO2)
Anti-diuretic horm
Anti-naturetic pro
Blood base excess
Urine sodium-pota
Volume of oxygen consumed (VO2)
Catecholamines, p
PH
End tidal fraction of oxygen (FetO2)
Urine osmolarity
Plasma lactate lev
Blood NO metabo
Glutathione perox
MDA
Nitrotyrosine
End tidal fraction of oxygen (FetCO2)
Plasma advanced
products and sup
Duration of inspiration and expiration
Haematocrit (Hct)
Haemoglobin con
Hypoxic ventilatory response (HVR)
Respiratory quotient (RQ)
Peripheral oxygen saturations (SpO2)
Arterial oxygen saturations (SaO2)
Arterial oxygen and carbon dioxide
partial pressure (PaO2) (PaCO2)
Arterial oxygen content
Alveolar-arterial PO2 difference
All the variables measured in the 13 accepted studies are listed. These have been subdivided into physiological systems. Ventilation AMS (acute mountain sickness)
and neurology
Other Sweat rates
Forced vascular conductance
Oesophageal temperature thresholds
for increasing forearm skin vascular
conductance
Oesophageal temperature thresholds
for increased sweat rate
Oesophageal temperature
Skin temperature
Urine volume
Plasma volume
Glomerular filtration rate (GFR)
Plasma potassium concentration (plasma K+)
Plasma sodium concentration (plasma Na+)
Plasma renin activity
Plasma aldosterone
Free water clearance
Adrenocorticotropic hormone (ACTH)
Anti-diuretic hormone (ADH)
Anti-naturetic protein (ANP)
Blood base excess
Urine sodium-potassium ratio (urine Na+/K+)
Catecholamines, plasma osmolarity
PH
Urine osmolarity
Plasma lactate levels
Blood NO metabolites
Glutathione peroxidase (GPX)
MDA
Nitrotyrosine
Plasma advanced oxidation protein
products and superoxide dismutase
Haematocrit (Hct)
Haemoglobin concentration (Hb conc) Alveolar ventilation (VA)
Volume of CO2 produced (VCO2) Volume of oxygen consumed (VO2)
End tidal fraction of oxygen (FetO2) All the variables measured in the 13 accepted studies are listed. These have been subdivided into physiological systems. volume found in a study was 0.9 L [12]. Two of the seven
studies reporting breathing frequency found it to be
higher in HH [17,18], whilst two others reported lower volume found in a study was 0.9 L [12]. Two of the seven
studies reporting breathing frequency found it to be
higher in HH [17,18], whilst two others reported lower this, the tidal volume was lower in HH in five out of six
studies where this was reported (Figure 3: Graph of tidal
volume) [12,17,18,21,22]. was reached.
bThe different altitudes were tested consecutively. So sometimes the exposure was 1 h at 3,000 or 1,700 m. cWhen PiO2 could not be calculated due to lack of information, the equivalent attitude estimated by the authors was given.
*P values were for three conditions; in recovery no P values unless stated in the discussion. re that of normal sea level but then were gradually made more hypoxic over the specified amount of time until the target hypoxic do ure was 1 h at 3,000 or 1,700 m.
ent attitude estimated by the authors was given was 1 h at 3,000 or 1,700 m.
ttitude estimated by the authors was given.
discussion This table describes all the features of the accepted studies.
aWhen the subjects entered the chamber, the environmental conditions were that of normal sea level but then were gradually made
was reached ted studies.
vironmental conditions were that of normal sea level but then were gradually made more hypoxic over the specified amount of time Ventilation The largest difference in tidal Table 2 Study design Table 2 Study design
Author and year
Type of outcome variable
Population
PiO2 of exposure (calculated by
PiO2 = (Pb-47) × FiO2 [4]) (mmHg)
Duration of exposure
(h:min)
Randomised
(Y, N)
Washout period
between trials (days:h)
NH HH
Basualto-Alarcon 2012
Ventilatory and
cardiovascular + exercise
7 men
3,000 mc
3,000 mc
00:15 builda, 00:10
N
7
Degache 2012
Postural stability
12 men
118 and 102
121 and 103
00:20–25 builda, 00:30b
Nil stated
<00:24
Hemmingsson 2009
Exhaled NO
6 men, 2 women
103 and 81
99 and 75
00:10 at each altitude
Y
<00:12
Loeppky 2005
Fluid balance
9 men
81
80
00:05 builda 10:00
Y
7
Loeppky 1997*
Ventilatory and cardiovascular
9 men
81
80
00:05 builda 10:00
Y
7
Miyagawa 2010
Ventilatory, cardiovascular
and sweat + exercise
7 men
93
97
00:30 builda only for HH 01:40
Y
>6
Naughton 1995
Haematological + exercise
9 subjects with chronic airflow
limitation (CAL) with 6 controls
1,829 and 2,438 mc
117 and 108
00:12 builda included 00:52:00
Y
00:02
Roach 1996
AMS and cardiovascular
9 men
4,564 mc
80
09:00
Y
7
Savourey 2003
Ventilatory and cardiovascular
18 men
4,500 mc
4,500 mc
00:10 builda, 00:30
Y
7
Savourey 2007
Ventilatory and cardiovascular
17 men, 1 woman
4,500 mc
4,500 mc
00:10 builda, 00:30
Y
14
Self 2011
AMS and cardiovascular
17 men and 3 women
7,620 mc
7,620 mc
00:05
N
<00:24
Tucker 1983
Mixed
11 men
82
80
00:15 builda included 02:20
N
Several weeks
Faiss 2013
Exhaled NO, ventilatory and
cardiovascular + exercise
10 men
99
101
24:00:00
Y
23
This table describes all the features of the accepted studies. aWhen the subjects entered the chamber, the environmental conditions were that of normal sea level but then were gradually made more hypoxic over the specified amount of time until the target hypoxic dose
was reached. bThe different altitudes were tested consecutively. So sometimes the exposure was 1 h at 3,000 or 1,700 m. cWhen PiO2 could not be calculated due to lack of information, the equivalent attitude estimated by the authors was given. PiO2 of exposure (calculated by
PiO2 = (Pb-47) × FiO2 [4]) (mmHg) Author and year
Type of outcome variable Coppel et al. Ventilation Extreme Physiology & Medicine (2015) 4:2 Page 6 of 20 Page 6 of 20 Table 3 Oxygenation and ventilation variables
Outcome
Author and year
Duration of
exposure
(h:min)
Hypobaric hypoxia
result [mean (SD)]
Normobaric
hypoxia result
[mean (SD)]
Difference
(HH −NH)
P value
Direction of
difference NH
compared to HH
VE (L/min)
Loeppky 1997
00:00
12.9
13.6
-
-
-
Savourey 2007
00:10 build, 00:05
10.49 (2.59)
10.14 (1.51)
-
>0.05
NS
Basualto-Alarcon
2012
00:15 build, 00:05
acclimatisation
10.5 (4.9)
10.3 (1.8)
0.2
-
-
Basualto-Alarcon
2012
00:15 build, 00:10
acclimatisation
35.7 (5.9)
39.7 (6.7)
-4
<0.05
NH > HH
Savourey 2003
00:10 build, 00:30
-
-
-
<0.02
NH > HH
Savourey 2007
00:10 build, 00:30
10.70 (1.93)
10.78 (1.93)
-
>0.05
NS
Faiss 2013
01:00
13.6 (1.8)
13.3 (3.3)
0.3
>0.05
NS
Miyagawa 2010a
01:05
69.2
65.6
-
>0.05
NS
Miyagawa 2010
01:10
70.5
65.7
-
>0.05
NS
Miyagawa 2010
01:20
73.9
70.9
-
>0.05
NS
Miyagawa 2010
01:40
75.3
77
-
>0.05
NS
Tucker 1983b
02:20 including
00:15 build
2.07
4.82
−2.75
-
-
Loeppky 1997
03:00
10.3
14.3
−4.0
<0.01
NH > HH
Loeppky 1997
06:00
10.6
12.7
−2.1
<0.05
NH > HH
Faiss 2013
08:00
11.8 (1.9)
14.9 (3.5)
−3.1
<0.1
NH > HH
-
10.7 (1.8)
12.2 (1.6)
−1.5
<0.05
NH > HH
-
12.7 (2.3)
14.2 (1.5)
−1.5
<0.1
NH > HH
Loeppky 1997
09:00
10.2
12.2
-
<0.05
NH > HH
Loeppky 1997
Recovery 12:00:00
9.2
10.1
-
>0.05
NS
VT (L)
Savourey 2007
00:10 buildc, 00:05
0.72 (0.25)
0.88 (0.22)
-
0.03
NH > HH
Basualto-Alarcon
2012
00:15 buildc, 00:05
acclimatisation
0.81 (0.36)
0.82 (0.21)
−0.01
-
-
Basualto-Alarcon
2012
00:15 buildc, 00:10
acclimatisation
1.85 (0.56)
1.91 (0.53)
−0.06
-
NH > HH
Savourey 2003
00:10 buildc, 00:30
-
-
-
<0.001
NH > HH
Savourey 2007
00:10 buildc, 00:30
0.83 (0.37)
0.86 (0.34)
-
>0.05
NS
Faiss 2013
01:00
0.88 (0.21)
0.89 (0.26)
−0.01
>0.05
NS
Tucker 1983b
02:20 including
00:15 buildc
106
152
-46
-
NH > HH
Faiss 2013
08:00
0.75 (0.21)
0.94 (0.3)
−0.19
<0.05
NH > HH
Faiss 2013
16:00
0.75 (0.23)
0.84 (0.24)
−0.9
<0.1
NH > HH
Loeppky 1997
10:00
-
-
-
>0.05
NS
Faiss 2013
24:00:00
0.86 (0.25)
0.95 (0.23)
−0.09
<0.05
NH > HH
Bf (cycles/min)
Savourey 2007
00:10 buildc, 00:05
15.73 (4.64)
12.24 (3.80)
-
0.03
HH > NH
Basualto-Alarcon
2012
00:15 buildc, 00:05
acclimatisation
13.3 (4.0)
13.4 (4.7)
−0.1
-
-
Basualto-Alarcon
2012
00:15 buildc, 00:10
acclimatisation
20.4 (5.4)
22.3 (7.4)
−1.9
<0.05
NH > HH
Savourey 2003
00:10 buildc, 00:30
-
-
-
<0.001
HH > NH
Savourey 2007
00:10 buildc, 00:30
14.77 (4.17)
13.76 (4.47)
-
>0.05
NS Table 3 Oxygenation and ventilation variables Coppel et al. Ventilation Extreme Physiology & Medicine (2015) 4:2 Page 7 of 20 Page 7 of 20 Table 3 Oxygenation and ventilation variables (Continued)
Faiss 2013
01:00
16.8 (3.4)
15.9 (4.2)
−0.9
>0.05
NS
Miyagawa 2010
01:05
31
31
-
>0.05
NS
Miyagawa 2010
01:10
32
34
-
>0.05
NS
Miyagawa 2010
01:20
34
36
-
>0.05
NS
Miyagawa 2010
01:40
38
42
-
>0.05
NS
Tucker 1983b
02:20 including
00:15 buildc
0.9
3.8
−2.9
-
NH > HH
Faiss 2013
08:00
16.8 (2.7)
17.1 (4.4)
−0.3
>0.05
NS
Loeppky 1997
10:00
-
-
-
>0.05
NS
Faiss 2013
16:00
16.1 (3)
15.8 (3.7)
−0.3
>0.05
NS
Faiss 2013
24:00:00
16.8 (3.8)
16.2 (3.8)
0.6
>0.05
NS
PetO2 (mmHg)
Savourey 2007
00:10 buildc, 00:05
72.5 (6.58)
79.56 (11.94)
-
0.08
Borderline NH >
HH
Savourey 2003
00:10 buildc, 00:30
-
-
-
>0.05
NS
Savourey 2007
00:10 buildc, 00:30
73.15 (7.16)
76.09 (11.61)
-
0.08
Borderline NH >
HH
Faiss 2013
01:00
66.4 (4.1)
62.3 (2.8)
-
4.1
NS
Tucker 1983b
02:20 including
00:15 buildc
−36.1
−32.6
−3.5
-
-
Faiss 2013
08:00
61.9 (6.0)
61.6 (2.2)
0.3
>0.05
NS
Loeppky 1997
10:00
-
-
-
>0.05
NS
Faiss 2013
16:00
65.0 (5.4)
62.7 (2.6)
2.3
>0.05
NS
Faiss 2013
24:00:00
65.6 (5.5)
65.6 (2.8)
0
>0.05
NS
PetCO2 (mmHg)
Savourey 2007
00:10 build, 00:05
44.09 (6.38)
48.87 (5.53)
-
0.05
Borderline NH >
HH
Savourey 2003
00:10 build, 00:30
-
-
-
>0.05
NS
Savourey 2007
00:10 build, 00:30
43.43 (6.02)
46.13 (6.61)
-
>0.05
NS
Faiss 2013
01:00
33.4 (2.5)
29.4 (2.4)
4
<0.1
HH > NH
Tucker 1983c
02:20 including
00:15 build
−2.8
−3.6
0.8
>0.05
NS
Faiss 2013
08:00
33.8 (2.1)
27.5 (1.3)
6.3
<0.01
HH > NH
Loeppky 1997
10:00
-
-
−1.6
<0.02
NH > HH
Faiss 2013
16:00
33.1 (1.3)
27.9 (0.9)
5.2
<0.01
HH > NH
Faiss 2013
24:00:00
30.8 (1.4)
26.5 (1.5)
4.3
<0.01
HH > NH
VA (alveolar
ventilation L/min)
Loeppky 1997
00:00
9.4
10.1
-
-
-
Loeppky 1997
03:00
7.2
10.5
−46%
<0.05
NH > HH
Loeppky 1997
06:00
7.6
9.1
-
<0.05
NH > HH
Loeppky 1997
09:00
7.6
9.2
-
<0.05
NH > HH
Loeppky 1997
Recovery 12:00:00
6.7
7.2
-
-
-
VCO2 ml/min
Loeppky 1997
00:00
295
333
-
-
-
Miyagawa 2010
01:05
2188
2108
-
>0.05
NS
Miyagawa 2010
01:10
2121
2007
-
>0.05
NS
Miyagawa 2010
01:20
2078
2060
-
>0.05
NS Table 3 Oxygenation and ventilation variables (Continued) Coppel et al. Ventilation Extreme Physiology & Medicine (2015) 4:2 Page 9 of 20 Page 9 of 20 values in HH [21,22] and there was no difference in the
remainder [12,13,15] (Figure 4: Graph of breathing fre-
quency). The only study that reported alveolar ventilation
Oxygenation
The peripheral oxygen saturations measured by pulse
oximetry (SpO2) were significantly lower in HH in two
Table 3 Oxygenation and ventilation variables (Continued)
Loeppky 1997
10:00
-
-
-
>0.05
NS
Faiss 2013
16:00
92 (2)
91 (2)
1
>0.05
NS
Faiss 2013
24:00:00
93 (2)
92 (1)
1
>0.05
NS
SaO2 (%)
Self 2011
00:05
-
-
-
0.005
NH > HH
Savourey 2007
00:10 buildc, 00:05
-
-
-
>0.05
NS
Savourey 2003
00:10 buildc, 00:30
85% (4)
88% (3)
−3
<0.05
NH > HH
Savourey 2007
00:10 buildc, 00:30
81.09% (7.76)
85.48% (5.63)
−4.39
0.07
Borderline NH >
HH
Miyagawa 2010
01:05
82
83
−1
>0.05
NS
Miyagawa 2010
01:10
81
82
−1
>0.05
NS
Miyagawa 2010
01:20
81
82
−1
>0.05
NS
Miyagawa 2010
01:40
82
81
1
>0.05
NS
Roach 1996
09:00
83% (1%)
83% (0.7%)
0
>0.05
NS
PaO2
Self 2011
00:05
-
-
-
0.004
HH > NH
Savourey 2007
00:10 buildc, 00:05
-
-
-
>0.05
NS
Savourey 2003
00:10 buildc, 00:30
6.38 (0.60)
6.90 (0.86)
−0.52
≤0.05
Borderline NH >
HH
Savourey 2007
00:10 buildc, 00:30
-
-
-
>0.05
NS
CAL subjects
Naughton 1995
00:52
-
-
−1.1
>0.05
NS
Control subjects
Naughton 1995
00:52
-
-
0.7
>0.05
NS
PaCO2
Self 2011
00:05
-
-
-
0.005
NH > HH
Savourey 2007
00:10 build, 00:05
-
-
-
>0.05
NS
Savourey 2003
00:10 build, 00:30
4.65 (0.54)
5.06 (0.46)
−0.41
≤0.05
Borderline NH >
HH
Savourey 2007
00:10 build, 00:30
46.3 (6.5)
52.2 (4.2)
−5.9
0.005
NH > HH
CAL subjects
Naughton 1995
00:52
-
-
0.3
>0.05
NS
Control subjects
Naughton 1995
00:52
-
-
−0.8
>0.05
NS
Tucker 1983b
02:20 including
00:15 build
−3.7
−5.6
1.9
-
-
Alveolar-arterial PO2
difference
CAL subjects
Naughton 1995
00:52
-
-
0.7
>0.05
NS
Control subjects
Naughton 1995
00:52
-
-
0.2
>0.05
NS
Arterial O2 content
Savourey 2007
00:10 buildc, 00:05
-
-
-
>0.05
NS
Savourey 2007
00:10 buildc, 00:30
-
-
-
>0.05
NS
This table lists all the values of the measured variables that relate to oxygenation and ventilation. Ventilation NS insignificant, hyphen no values given, CAL Chronic Airflow Limitation. aExercise started at 01:00. bDifference from controls. cWhen the subjects entered the chamber, the environmental conditions were that of normal sea level but then were gradually made more hypoxic over the
specified amount of time until the target hypoxic dose was reached. Table 3 Oxygenation and ventilation variables (Continued) This table lists all the values of the measured variables that relate to oxygenation and ventilation. NS insignificant, hyphen no values given, CAL Chronic Airflow Limitation. aExercise started at 01:00. bDifference from controls. cWhen the subjects entered the chamber, the environmental conditions were that of normal sea level but then were gradually made more hypoxic over the
specified amount of time until the target hypoxic dose was reached. bDifference from controls. cWhen the subjects entered the chamber, the environmental conditions were that of normal sea level but then were gradually made more hypoxic over the
specified amount of time until the target hypoxic dose was reached. values in HH [21,22] and there was no difference in the
remainder [12,13,15] (Figure 4: Graph of breathing fre-
quency). The only study that reported alveolar ventilation
found that it was higher in NH than HH [13]. Ventilation Extreme Physiology & Medicine (2015) 4:2 Page 8 of 20 Page 8 of 20 Table 3 Oxygenation and ventilation variables (Continued)
Miyagawa 2010
01:40
2021
2082
-
>0.05
NS
Loeppky 1997
03:00
216
330
-
<0.05
NH > HH
Loeppky 1997
06:00
227
296
-
<0.05
NH > HH
Loeppky 1997
09:00
235
302
−67
<0.05
NH > HH
Loeppky 1997
Recovery 12:00
241
267
-
-
-
VO2 consumed
Loeppky 1997
00:00
329
340
-
-
-
Miyagawa 2010
01:05
1709
1611
-
>0.05
NS
Miyagawa 2010
01:10
1783
1637
-
>0.05
NS
Miyagawa 2010
01:20
1826
1748
-
>0.05
NS
Miyagawa 2010
01:40
1836
1840
-
>0.05
NS
Loeppky 1997
03:00
250
361
-
<0.05
NH > HH
Loeppky 1997
06:00
262
319
-
<0.05
NH > HH
Loeppky 1997
09:00
278
326
-
<0.05
NH > HH
Loeppky 1997
12:00
301
291
-
-
-
FetO2
Savourey 2007
00:10 buildc, 00:05
-
-
-
>0.05
NS
Savourey 2003
00:10 buildc, 00:30
-
-
-
<0.00001
HH > NH
Savourey 2007
00:10 buildc, 00:30
-
-
-
>0.05
NS
FetCO2
Savourey 2007
00:10 build, 00:05
-
-
-
>0.05
NS
Savourey 2003
00:10 build, 00:30
-
-
-
<0.00001
HH > NH
Savourey 2007
00:10 build, 00:30
-
-
-
>0.05
NS
Duration of
inspiration/s
Savourey 2007
00:10 buildc, 00:05
1.94 (0.65)
2.99 (0.98)
-
0.01
NH > HH
Savourey 2007
00:10 buildc, 00:30
2.40 (1.25)
3.00 (1.16)
-
>0.05
NS
Duration of expiration/s
Savourey 2007
00:10 buildc, 00:05
2.09 (0.87)
1.98 (0.84)
-
>0.05
NS
Savourey 2007
00:10 buildc, 00:30
-
-
-
>0.05
NS
Hypoxic ventilatory
response (HVR) 1%−1
Savourey 2007
00:10 buildc, 00:05
−0.05
0.03
-
>0.05
NS
Savourey 2007
00:10 buildc, 00:30
−0.09
−0.07
-
>0.05
NS
SpO2 (%)
Savourey 2007
00:10 buildc, 00:05
83.03 (4.49)
87.11 (4.81)
−4.08
<0.05
NH > HH
Basualto-Alarcon
2012
00:15 buildc, 00:05
acclimatisation
91.6 (4.2)
89.1 (3.8)
2.5
<0.05
HH > NH
Basualto-Alarcon
2012
00:15 buildc, 00:10
acclimatisation
85.3 (3.8)
86.0 (1.7)
−0.7
-
-
Savourey 2003
00:10 buildc, 00:30
-
-
-
<0.05
NH > HH
Savourey 2007
00:10 buildc, 00:30
82.49 (4.39)
85.50 (4.84)
−2.99
0.04
NH > HH
Faiss 2013
01:00
93 (1)
90 (3)
3
>0.05
NS
Tucker 1983b
02:20 including
00:15 buildc
−13.2
−13.5
0.3
>0.05
NS
Faiss 2013
08:00
91 (3)
91 (3)
0
>0.05
NS
Roach 1996
09:00
83% (1%)
83% (0.7%)
0
>0.05
NS
Loeppky 2005
10:00
82%
83%
−1%
>0.05
NS Table 3 Oxygenation and ventilation variables (Continued) Coppel et al. Oxygenation Extreme Physiology & Medicine (2015) 4:2 Page 11 of 20 Table 4 Cardiovascular variables (Continued)
BP systolic (Torr)
Faiss 2013
01:00
124 (9)
129 (13)
−5
>0.05
NS
Tucker 1983b
02:20 including
00:15 builda
−1
5
−6
NH > HH
Faiss 2013
08:00
124 (9)
123 (7)
1
>0.05
NS
Faiss 2013
16:00
121 (9)
118 (9)
3
>0.05
NS
Faiss 2013
24:00:00
131 (10)
129 (9)
2
>0.05
NS
This table lists all the values of the measured variables that relate to the cardiovascular system. NS insignificant. aWhen the subjects entered the chamber, the environmental conditions were that of normal sea level but then were gradually made more hypoxic over the
specified amount of time until the target hypoxic dose was reached. bDifference from controls. Table 4 Cardiovascular variables (Continued) Table 4 Cardiovascular variables (Continued) NS insignificant. aWhen the subjects entered the chamber, the environmental conditions were that of normal sea level but then were gradually made more hypoxic over the
specified amount of time until the target hypoxic dose was reached. Oxygenation The peripheral oxygen saturations measured by pulse
oximetry (SpO2) were significantly lower in HH in two
out of three short studies [17,18]. One study found that Coppel et al. Extreme Physiology & Medicine (2015) 4:2 Page 10 of 20 Page 10 of 20 Table 4 Cardiovascular variables
Outcomes
Author and year
Duration of
exposure (h:min)
Hypobaric hypoxia
result [mean (SD)]
Normobaric hypoxia
result [mean (SD)]
Difference
(HH −NH)
P value
Direction of
difference NH
compared to HH
Hypoxic cardiac
response (HCR)
bpm %−1
Savourey 2007
00:10 builda, 00:05
−0.61
−0.63
0.02
>0.05
NS
Savourey 2007
00:10 builda, 00:30
−0.52
−0.79
0.27
>0.05
NS
LF/HF%
Basualto-Alarcon 2012
00:15 builda, 00:10
acclimatisation
1.96 (2.6)
1.28 (0.92)
0.68
<0.05
HH > NH
HR
Self 2011
00:01
104.9 (14.3)
96.6 (14.6)
8.3
<0.05
HH > NH
Self 2011
00:04
-
-
-
>0.05
NS
Basualto-Alarcon 2012
00:15 builda, 00:05
acclimatisation
61 (9)
62 (6)
−1
-
-
Basualto-Alarcon 2012
00:15 builda, 00:10
acclimatisation
129 (23)
134 (16)
−5
<0.05
NH > HH
Savourey 2007
00:10 builda, 00:05
70.32 (9.91)
69.62 (9.95)
>0.05
NS
Savourey 2003
00:10 builda, 00:30
-
-
-
<0.05
HH > NH
Savourey 2007
00:10 builda, 00:30
69.50 (12.07)
70.67 (12.07)
-
>0.05
NS
Faiss 2013
01:00
62 (8)
63 (10)
−1
>0.05
NS
Miyagawa 2010
01:05
144
137
-
>0.05
NS
Miyagawa 2010
01:10
150
146
-
>0.05
NS
Miyagawa 2010
01:40
166
164
-
>0.05
NS
Tucker 1983a
02:20 including
00:15 builda
10.4
3.6
6.8
-
HH > NH
Faiss 2013
08:00
68 (13)
69 (13)
−1
>0.05
NS
Faiss 2013
16:00
61 (10)
66 (7)
−5
>0.05
NS
Faiss 2013
24:00
65 (9)
71 (10)
−6
>0.05
NS
Stroke volume
Miyagawa 2010
01:05
105
107
−2
>0.05
NS
Miyagawa 2010
01:10
113
126
−13
>0.05
NS
Miyagawa 2010
01:40
116
124
−8
>0.05
NS
Cardiac output
Miyagawa 2010
01:05
15.3
14.7
>0.05
NS
Miyagawa 2010
01:10
17.1
18.2
>0.05
NS
Miyagawa 2010
01:40
19.4
20.1
>0.05
NS
Mean BP
Miyagawa 2010
01:05
112
107
>0.05
NS
Miyagawa 2010
01:10
108
107
>0.05
NS
Miyagawa 2010
01:40
100
99
>0.05
NS Table 4 Cardiovascular variables Coppel et al. This table lists all the values of the measured variables that relate to AMS and neurology.
NS insignificant. g
*P value calculated including hypobaric normoxia. g
*P value calculated including hypobaric normoxia. NS insignificant.
*P value calculated including hypobaric normoxia. ant.
culated including hypobaric normoxia. Oxygenation bDifference from controls Table 5 AMS and neurology variables gy
Outcome
Author and
year
Duration of
exposure (h:min)
Hypobaric hypoxia
result [mean (SD)]
Normobaric hypoxia
result [mean (SD)]
Difference
(HH −NH) P value Direction of differenc
NH compared to HH
Length of centre
of pressure trajectory
in Y axis
Eyes open 1,700 m
Degache 2012
00:30
114.2 (38.8)
129.5 (53.3)
-15.3
-
NH > HH
Eyes closed 1,700 m Degache 2012
00:30
127.2 (54.9)
87.7 (44.8)
39.5
-
HH > NH
Dual task 1,700 m
Degache 2012
00:30
128.7 (87.1)
79.9 (30.3)
48.8
-
HH > NH
Romberg’s index
1,700 m
Degache 2012
00:30
1.35 (0.19)
1.42 (0.34)
-0.07
-
-
Eyes open 3,000 m
Degache 2012
00:30
123.1 (22.6)
127.2 (41.5)
-4.1
-
-
Eyes closed 3,000 m Degache 2012
00:30
104.7 (27.0)
89.1 (39.9)
15.6
-
HH > NH
Dual task 3,000 m
Degache 2012
00:30
91.9 (22.4)
82.4 (30.4)
9.5
-
HH > NH
Romberg’s index
3,000 m
Degache 2012
00:30
1.33 (0.22)
1.39 (0.29)
-0.06
-
-
Variance of speed
of CoP
Eyes open 1,700 m
Degache 2012
00:30
111.0 (56.2)
151.4 (30.2)
-40.4
-
NH > NH
Eyes closed 1,700 m Degache 2012
00:30
111.0 (58.8)
149.9 (31.5)
-38.9
-
NH > HH
Dual task 1,700 m
Degache 2012
00:30
112.1 (57.7)
151.1 (31.7)
-39
-
NH > HH
Romberg’s index
1,700 m
Degache 2012
00:30
0.98 (0.09)
0.99 (0.03)
-0.01
-
-
Eyes open 3,000 m
Degache 2012
00:30
150.5 (42.3)
160.8 (14.0)
-10.3
-
NH > HH
Eyes closed 3,000 m Degache 2012
00:30
142.9 (40.8)
158.6 (13.7)
-15.7
-
NH > HH
Dual task 3,000 m
Degache 2012
00:30
143.4 (39.2)
160.1 (15.0)
-16.7
-
NH > HH
Romberg’s index
3,000 m
Degache 2012
00:30
0.95 (0.11)
0.99 (0.02)
-0.04
-
-
Lake Louise AMS
scores
Self 2011
00:01
-
-
2.36
>0.05
NS
Self 2011
00:04
-
-
-4.89
>0.05
NS
Roach 1996
09:00
-
-
-
<0.01
HH > NH
Loeppky 2005*
10:00
-
-
-
<0.001
HH > NH
Thi t bl
li t
ll th
l
f th
d
i bl
th t
l t
t
AMS
d
l Outcome
Author and
year
Duration of
exposure (h:min)
Hypobaric hypoxia
result [mean (SD)]
Normobaric hypoxia
result [mean (SD)]
Difference
(HH −NH) P value Direction of difference
NH compared to HH Coppel et al. Oxygenation Extreme Physiology & Medicine (2015) 4:2 Page 12 of 20 Page 12 of 20 Table 6 Additional physiological variables
Outcome
Author and year
Duration of
exposure (h:min)
Hypobaric
hypoxia result
[mean (SD)]
Normobaric
hypoxia result
[mean (SD)]
Difference
(HH −NH)
P value
Direction of
difference NH
compared to H
Exhaled NO (PE NO)
Hemmingsson 2009
00:10 at each
ascending altitude
-
-
33% mean
reduction
(at 5,000 m)
0.002
NH > HH
Faiss 2013
01:00
9.5 (5.0)
14.9 (9.2)
−5.4
<0.01
NH > HH
Faiss 2013
08:00
8.8 (5.3)
14.1 (7.4)
−5.3
<0.01
NH > HH
Faiss 2013
16:00
7.9 (4.5)
14.7 (8.6)
−6.8
<0.01
NH > HH
Faiss 2013
24:00:00
8.9 (5.4)
15.7 (8.7)
−5.8
<0.01
NH > HH
RQ
Self 2011
00:05
2.37 (0.53)
1.41 (0.15)
0.96
0.005
HH > NH
Forced vascular conductance
Miyagawa 2010
01:00-01:40
-
-
-
>0.05
NS
Sweat rate
Miyagawa 2010
01:00-01:40
-
-
-
>0.05
NS
Oesophageal temperature
thresholds for increasing
forearm skin vascular
conductance
Miyagawa 2010
01:00-01:40
-
-
-
>0.05
NS
Oesophageal temperature
thresholds for increasing
sweat rate
Miyagawa 2010
01:00-01:40
-
-
-
>0.05
NS
Oesophageal temperature
Miyagawa 2010
01:05
36.63
36.61
0.02
>0.05
NS
Miyagawa 2010
01:10
37.12
37.11
0.01
>0.05
NS
Miyagawa 2010
01:40
37.95
37.96
−0.01
>0.05
NS
Skin temperature
Miyagawa 2010
01:05
33.37
33.47
−0.1
>0.05
NS
Miyagawa 2010
01:10
33.35
33.43
−0.08
>0.05
NS
Miyagawa 2010
01:40
34.44
34.59
−0.15
>0.05
NS
Urine vol (ml)
Loeppky 2005a
10:00
-
-
-
0.005
HH > NH
Tucker 1983b
02:20 including
00:15 buildd
−1.6
0.1
−1.7
-
NH > HH
Plasma volume
Miyagawa 2010
01:00-01:40
-
-
-
>0.05
NS
Loeppky 2005
10:00
-
-
−6%
0.002**
HH > NH
GFR
Loeppky 2005
10:00
-
-
-
>0.05
NS
Plasma K+
Loeppky 2005
10:00
-
-
-
0.003
NH > HH
Plasma Na+
Loeppky 2005
10:00
-
-
-
0.006
NH > HH
Plasma renin activity (PRA)
Loeppky 2005a
10:00
-
-
-
<0.05
HH > NH Table 6 Additional physiological variables RQ Coppel et al. Oxygenation Extreme Physiology & Medicine (2015) 4:2 Page 13 of 20 Table 6 Additional physiological variables (Continued)
Plasma aldosterone
Loeppky 2005a
10:00
-
-
-
<0.001
NH > HH
Free water clearance (CH2O)
Loeppky 2005a
10:00
-
-
-
<0.05
HH > NH
ACTH
Loeppky 2005a
10:00
-
-
-
0.18
NS
ADH
Loeppky 2005
10:00
-
-
-
>0.05
NS
ANP
Loeppky 2005*
10:00
-
-
-
0.97
NS
Blood base excess
Loeppky 2005
10:00
-
-
-
>0.05
NS
Urine Na+/K+
Loeppky 2005
10:00
-
-
-
0.7
NS
Catecholamines
Miyagawa 2010
01:00-01:40
-
-
-
>0.05
NS
Loeppky 2005
10:00
-
-
-
0.43
NS
Haematocrit
Miyagawa 2010
01:00-01:40
-
-
-
>0.05
NS
Tucker 1983b
02:20 including
00:15 buildd
0.7
0.7
0
NS
Haemoglobin concentration
Savourey 2007
00:10 buildd, 00:05
-
-
-
>0.05
NS
Savourey 2007
00:10 buildd, 00:30
-
-
-
>0.05
NS
Miyagawa 2010
01:00-01:40
-
-
-
>0.05
NS
Plasma osmolarity
(mOsm)
Miyagawa 2010
01:00-01:40
-
-
-
>0.05
NS
Tucker 1983b
02:20 including
00:15 buildd
−0.4
−1
0.6
-
HH > NH
pH
Savourey 2003
00:10 buildd, 00:30
7.46 (SEM 0.03)
7.44 (SEM 0.02)
0.02
≤0.05
Borderline HH > NH
Savourey 2007
00:10 buildd, 00:30
7.45 (0.04)
7.44 (0.04)
0.01
0.02
HH > NH
CAL subjects
Naughton 1995
00:52
-
-
−0.02
>0.05
NS
Control subjects
Naughton 1995
00:52
-
-
0.01
>0.05
NS
Tucker 1983d
02:20 including
00:15 buildd
0.047
0.015
0.032
-
HH > NH
Faiss 2013
24:00:00
-
-
-
<0.01
NH > HH
Urine osmolarity (mOsm)
Tucker 1983c
02:20 including
00:15 buildb
15
0.1
14.9
-
HH > NH
Lactate mmol/kgH2O
Miyagawa 2010
01:00-01:40
-
-
-
>0.05
NS Table 6 Additional physiological variables (Continued) Coppel et al. Oxygenation Extreme Physiology & Medicine (2015) 4:2 Page 14 of 20 Table 6 Additional physiological variables (Continued) Table 6 Additional physiological variables (Continued) Table 6 Additional physiological variables (Continued)
Blood NO metabolites
Faiss 2013
01:00
31.6 (19.6)
27.7 (7.3)
3.9
<0.01
HH > NH
Faiss 2013
08:00
28.1 (18.9)
32.7 (9.7)
−4.6
<0.01
NH > HH
Faiss 2013
16:00
24.2 (16.3)
30.2 (7.1)
−6
<0.01
NH > HH
Faiss 2013
24:00
22.85 (16.2)
28.9 (6.9)
−6.05
<0.01
NH > HH
GPX (% baseline)
Faiss 2013
01:00
114 (26)
111 (30)
3
>0.05
NS
Faiss 2013
08:00
85 (27)
123 (23)
−37
>0.05
NS
Faiss 2013
16:00
105 (43)
107 (21)
−2
>0.05
NS
Faiss 2013
24:00
103 (43)
107 (21)
−4
>0.05
NS
MDA (% baseline)
Faiss 2013
01:00
117 (40)
92 (36)
25
>0.05
NS
Faiss 2013
08:00
103 (62)
111 (35)
−8
>0.05
NS
Faiss 2013
16:00
111 (56)
116 (55)
−5
>0.05
NS
Faiss 2013
24:00
108 (52)
97 (51)
11
>0.05
NS
Nitrotyrosine (% baseline)
Faiss 2013
01:00
86 (16)
105 (26)
−19
>0.05
NS
Faiss 2013
08:00
77 (35)
75 (37)
2
>0.05
NS
Faiss 2013
16:00
91 (20)
98 (16)
−7
>0.05
NS
Faiss 2013
24:00
75 (40)
87 (25)
−12
>0.05
NS
Plasma advanced oxidation
protein products
Faiss 2013
01:00
120%
13%
107%
-
NH > HH
Faiss 2013
24:00
260%
88%
172%
-
NH > HH
Superoxide dismutase
Faiss 2013
24:00
-
-
37%
-
NH > HH
This table lists all the values of all other the measured physiological variables. NS insignificant, SEM Standard Error of the Mean. aMeasured 2 h after exposure. bDifference from controls. cSubjects were gradually exposed increasing levels of hypoxia over the stated time until the target hypoxic dose was reached. dWhen the subjects entered the chamber, the environmental conditions were that of normal sea level but then were gradually made more hypoxic over the
specified amount of time until the target hypoxic dose was reached. *P value calculated including hypobaric normoxia. **P value calculated including hypobaric normoxia and after 3 h. Carbon dioxide (CO2) clearance Each data
point represents data obtained from a study and the number refers to the time point. If the data point is in the green area, the minute ventilation was
found to be higher in HH but if in the blue area, the minute ventilation was found to be lower in HH. Carbon dioxide (CO2) clearance the saturations were 4.08% lower in HH [18]. However,
no differences were found in all five of the long studies
[12-14,16,22]. The arterial blood saturations (SaO2) were
lower in HH in all three short studies [6,17,18] but not
in the two longer studies [15,16]. Arterial partial pres-
sures of oxygen (PaO2) was lower in NH in one study
[6], higher in NH in one study [17], and no different in
two studies [18,20] (Table 3: Oxygenation and ventila-
tion). Only the two studies by Savourey et al. [17,18]
measured the end tidal fractions of O2 and these report
discordant results. In 2003, Savourey et al. [17] found
the end tidal fractions of O2 to be higher (P < 0.00001)
in HH than NH; however, in their 2007 [18] study follow-
ing the same protocols, no difference was demonstrated
(P > 0.05). )
In three out of five studies, there was no difference in
the end tidal partial pressure of CO2 (PETCO2) between
HH and NH [17,18,22]; however, one study [12] found it
to be higher in HH and one found it to be lower in HH
[13]. In four out of five studies, the PaCO2 levels were
lower [11,17,13] or the same [20] in HH compared to
NH. For example, one study found the difference in
PaCO2 to be as large as 5.9 hPa lower in HH [18]. Two
studies measured the end tidal fractions of CO2 [17,18]. In 2003, Savourey et al. [17] found the end tidal fractions
of CO2 to be higher (P < 0.00001) in HH than NH,
whereas in the same group's 2007 study [18] (following
the same protocols), no difference was demonstrated
(P > 0.05). Coppel et al. Extreme Physiology & Medicine (2015) 4:2 Page 15 of 20 Figure 2 Graph of minute ventilation. Graph to show the difference in minute ventilation between the two environments over time. Each data
point represents data obtained from a study and the number refers to the time point. If the data point is in the green area, the minute ventilation was
found to be higher in HH but if in the blue area, the minute ventilation was found to be lower in HH. Figure 2 Graph of minute ventilation. Graph to show the difference in minute ventilation between the two environments over time. Acute mountain sickness and neurology to NH. Subjects deviated from the midline more in HH
than NH, and the speed with which movements oc-
curred to correct their posture was slower. Four studies were found relating to acute mountain sick-
ness (AMS) and neurological symptoms and signs, two
of which were long studies [14,16] and two of which
were short studies [6,23] (Table 5: AMS and neurology). Two out of three studies [14,16] that measured AMS
scores found that AMS was significantly worse in HH
than in NH. Only one study [23] measured postural sta-
bility, which was significantly reduced in HH compared Cardiovascular variables If the data point is in the green area, the breathing
frequency was found to be higher in HH but if in the blue area, the breathing frequency was found to be lower in HH. Cardiovascular variables (Figure 5: Graph of heart rates). Three studies measured
blood pressure. Two found no difference in blood pressure
between environments [12,15]; however, one other found
it to be lower in HH than in NH [22]. Sympathetic drive,
measured by a specific electrocardiogram (ECG) recorder
of low- and high-frequency components of heart rate, was
only investigated in one study [21] and was found to be
higher in HH than in NH. Seven studies were identified that reported physiological
variables relating to the cardiovascular system. Three of
these were long studies [12,15,22] and four were short
studies [6,17,18,21] (Table 4: Cardiovascular variables). All seven studies measured heart rate (HR). Three found
HR to be higher in HH [6,17,22], one found it to be lower
[21], and the others found no differences [12,15,18] Figure 3 Graph of tidal volume. Graph to show the difference in tidal volume between the two environments over time. Each data point
represents data obtained from a study and the number refers to the time point. If the data point is in the green area, the tidal volume was found
to be higher in HH but if in the blue area, the tidal volume was found to be lower in HH. Figure 3 Graph of tidal volume. Graph to show the difference in tidal volume between the two environments over time. Each data point
represents data obtained from a study and the number refers to the time point. If the data point is in the green area, the tidal volume was found
to be higher in HH but if in the blue area, the tidal volume was found to be lower in HH. Coppel et al. Extreme Physiology & Medicine (2015) 4:2 Page 16 of 20 Figure 4 Graph of breathing frequency. Graph to show the difference in breathing frequency between the two environments over time. Each
data point represents data obtained from a study and the number refers to the time point. If the data point is in the green area, the breathing
frequency was found to be higher in HH but if in the blue area, the breathing frequency was found to be lower in HH. Figure 4 Graph of breathing frequency. Graph to show the difference in breathing frequency between the two environments over time. Each
data point represents data obtained from a study and the number refers to the time point. Oxygenation and ventilation parameters The majority of studies included data relating to oxygenation
and ventilation. Oxygen saturations measured from pulse
oximetry (SpO2) and arterial blood gases correlated with each
other. During short-term exposure, oxygen saturations were
lower in HH [6,17]. This was not the case for long-term
exposure, perhaps as the initial process of acclimatisation to
hypoxia had occurred [18,20]. PaO2 did not differ at any stage. Only two studies [12,19] measured exhaled NO levels
and both found that they were lower in HH than in
NH. The greatest difference in exhaled NO found in
these papers was 6.8 mmHg [12]. Additionally, only
one study measured systemic NO and the authors
found it was lower in HH [12]. The same study was the
only study to measure plasma-advanced oxidation pro-
tein products and superoxide dismutase. These were
higher in HH [12]. Only one study [6] measured the re-
spiratory quotient (RQ) and found it was higher in HH
than NH. A decreased minute ventilation and alveolar ventila-
tion was seen in HH. This finding is in accord with the
smaller tidal volumes demonstrated in HH; however,
breathing frequencies varied [12,13,17,18,21,22]. Despite
the lower SpO2 in HH initially, oxygen saturations were
maintained at the same level in both environments over
longer time periods. Interestingly, despite the reduction
in ventilation, PETCO2 levels did not change significantly
[12,13,17,18,22]. Notably, Savourey et al. found different results in end
tidal fractions of both O2 and CO2 between their two
studies in 2003 [17] and 2007 [18]. This not only makes
interpretation of their results very challenging but also
highlights issues concerning reliability of studies (as
discussed in Study quality section). Additional physiological variables Six studies [6,12,14,15,19,22] researched a variety of other
physiological variables (Table 6: Additional physiological
variables). In three out of five of these studies [17,18,22],
the plasma pH was higher in HH than NH; however, one of Figure 5 Graph of heart rates. Graph to show the difference in heart rate between the two environments over time. Each data point represents
data obtained from a study and the number refers to the time point. If the data point is in the green area, the heart rate was found to be higher in
HH but if in the blue area, the heart rate was found to be lower in HH. Figure 5 Graph of heart rates. Graph to show the difference in heart rate between the two environments over time. Each data point represents
data obtained from a study and the number refers to the time point. If the data point is in the green area, the heart rate was found to be higher in
HH but if in the blue area, the heart rate was found to be lower in HH. Coppel et al. Extreme Physiology & Medicine (2015) 4:2 Page 17 of 20 Significant differences were demonstrated in variables
relating to ventilation, NO, fluid retention, and in fac-
tors relating to AMS. For other variables, there was no
consistent pattern across the reviewed studies. the other papers found the pH to be higher in NH [12]
(Figure 6: Graph of pH). The greatest difference in pH
found in a study was 0.032 [22]. Urine osmolarity was mea-
sured in only one study and was significantly higher in HH
[22]. Additionally, the same study was the only one to
measure the volume of urine produced and found it to be
lower during HH exposure [22]. One study also measured
K+, Na+ and aldosterone concentrations in plasma and
found they were higher in NH [14]. In recovery post-
hypoxic exposure, authors in [14] found that the urine vol-
ume, plasma renin activity and free water clearance were
higher whilst aldosterone was lower after HH than NH. This was the only study to measure these variables. Mechanisms for results
h
h
l Plasma pH appears to be higher in HH than NH
[17,18,22]. The study [20] that did not find significant
differences in pH between the two conditions was con-
ducted at 1,829 and 2,438 m whereas the elevated pH
values were from 4,500 m, suggesting that the differ-
ences between the two conditions may be more pro-
nounced at higher altitudes. Elevated pH in HH is
surprising in the context of the finding that ventilation is
reduced under these conditions. Many hypothetical mechanisms have been proposed for
the effect of low barometric pressure on physiology. These
include intravascular bubble formation, increased alveolar
deadspace, altered fluid permeability, changes in chemo-
sensitivity, and a mismatch in ventilation and perfusion
[13,16,17]. Although pressure may be the principle con-
founder between the two scenarios, we must also reflect
that other factors may differ between HH and NH, thus
impacting participant's physiology. For example, the
laboratory-based components of the studies reviewed were
performed between 22°C and 25°C, a temperature likely to
be far warmer that experiences at 4,000 m in a field la-
boratory. Such differences in ambient temperature may
alter physiological mechanisms such as the degree of per-
ipheral vasoconstriction, NO metabolism or the produc-
tion of reactive oxidative species [24]. Exhaled and systemic NO levels were lower in HH
[12,19]. Faiss et al. [12] found increased oxidative stress
in HH and attributed the systemic differences in NO
bioavailability to this. Many other physiological variables were measured;
however, most of these were reported in only a single
study. Thus, it is difficult to make conclusions without
verification from other studies, and we have not consid-
ered these further. As highlighted, the duration of the hypoxic exposure
impacts on the results obtained. Different physiological
systems will have different response rates for adaptation
to hypoxia [25]. For some physiological parameters, the
short study durations may not be long enough for dif-
ferences between NH and HH to be elicited. Studies
reporting repeated measures over time provide a win-
dow on this phenomenon. For example, in the 1997
study by Loeppky et al. [13], where no differences in
minute ventilation were reported after 30 or 60 min of
hypoxic exposure, significant differences were evident
after 3 h of exposure. Additionally, Savourey et al. [17,18] initially found lower PETO2 and PETCO2 in HH
than NH but then no difference in prolonged exposure. Summary of physiological findings We believe this is the first systematic review com-
paring the effect of NH and HH on human physiology. Figure 6 Graph of pHs. Graph to show the difference in pH between the two environments over time. Each data point represents data obtained
from a study and the number refers to the time point. If the data point is in the green area, the pH was found to be higher in HH but if in the blue
area, the pH was found to be lower in HH. Figure 6 Graph of pHs. Graph to show the difference in pH between the two environments over time. Each data point represents data obtained
from a study and the number refers to the time point. If the data point is in the green area, the pH was found to be higher in HH but if in the blue
area, the pH was found to be lower in HH. Coppel et al. Extreme Physiology & Medicine (2015) 4:2 Coppel et al. Extreme Physiology & Medicine (2015) 4:2 Page 18 of 20 AMS and neurology variables Mountain sickness is an area of research that has
attracted much interest and consequently the multiple
variables that combine to make the Lake Louise AMS
scoring system are commonly collected in these studies. In two of four studies, AMS severity was found to in-
crease in HH compared with NH [14,16]. Furthermore,
in the one study that found no differences between envi-
ronments [6], participants only had a 5-min exposure
time, as opposed to 9- and 10-h exposures in other stud-
ies. Additionally,
postural
stability
was
significantly
worse in HH [23]. The authors suggest that visual con-
trast sensitivity is lower in HH than in NH and that this
may contribute to the postural stability [23]. Cardiovascular variables Failure to account for the PH2O leads to an overesti-
mate of the hypoxic dose in NH, such that incorrect par-
tial pressure of inspired oxygen (PiO2) may be attained
[11]. In one study [6], a NH exposure equivalent to
7,620 m was described; however, the conditions were in
fact closer to 7,010 m once pH2O was accounted for
[11]. We have emphasised these differences by calculat-
ing, where possible, the PiO2 in the different hypoxic
conditions (Table 2: Study design). We found the differ-
ences in PiO2 to be as much as 4 mmHg. The level of
CO2 in the test environment was a potential source of
error. Basualto-Alarcon et al. [21] highlight this issue in
stating that different gas inflow rates into each hypoxic
system allow different levels of CO2 accumulation. Add-
itionally, they state that their NH environment may have
been more hypercapnic because it had half the total vol-
ume of the hypobaric chamber. These control group
contrast issues will either enhance or diminish the effect
size and therefore the difference between HH and NH. This
may
be
of
particular
relevance
to
ventilator
variables. The majority of these variables were similar for HH and
NH. Heart rate and blood pressure results were incon-
sistent, although some data suggests that heart rate may
be raised initially in HH [6,12,15,17,18,21,22]. Mechanisms for results
h
h
l This may be because during HH exposure, the ambient Study quality
Th
h The search results reveal several issues relating to study
design. Very few studies state the reliability of their mea-
surements or performed a sample size calculation. It is
therefore difficult to evaluate if they are adequately pow-
ered to identify a real difference between conditions,
should such a difference exist. Given that differences in
physiological responses between NH and HH conditions
are likely quite small, large sample sizes would likely be
required to avoid type II (false negative) errors. Add-
itionally, by performing statistical analysis on a large
number of variables over many time points, the risk of
type I (false positive) errors increases. Coppel et al. Extreme Physiology & Medicine (2015) 4:2 Page 19 of 20 the small sample sizes may be associated with an increased
likelihood of type 2 errors. partial pressure of nitrogen (PN2) is initially lower than
the body's and therefore nitrogen (N2) initially diffuses
from the tissues to the alveoli [5]. Until this equilibrium
is achieved, the arterial oxygen content, PAO2, and the
arterial carbon dioxide content, PACO2, are lowered as
a result of the relatively higher PAN2 in HH than NH. As mentioned, the studies were heterogeneous by de-
sign. For example, they differed in regard to the subjects'
prior exposure to altitude. In two of the studies reviewed
[16,22], the subjects lived between 1,500 and 1,600 m
and so may have been partially acclimatised to high alti-
tude. It is not clear whether the same effects would be
seen in partially and not acclimatised subjects. Loeppky et al. [13] also suggests that an initial increase
in CO2 produced in HH compared to NH might be due
to microbubble formation similar to the nitrogen bends
in divers. This emphasises the importance of study
duration on physiological response and the problems
inherent in comparing studies of different hypoxia ex-
posure times. Finally, the self-reported nature of AMS scores could be
associated with inconsistent responses from participants. In the study by Self et al. [6], there was a disparity between
post-hypoxia interview responses and the responses dur-
ing hypoxic exposure. There is no gold standard method
for these types of studies and so there is much variability
due to the methodology employed. If there are indeed differences between HH and NH,
at what equivalent altitude do they become apparent? Conclusions
W We present an overview of the current available litera-
ture regarding crossover studies relating to the different
effects of HH and NH on human physiology. This sys-
tematic review is the first to compare the effects of a
NH and HH environment on human physiology. Several
studies reported a number of variables (e.g. minute ven-
tilation and NO levels) that were different between the
two conditions, lending support to the notion that true
physiological difference are indeed present. However, the
presence of confounding factors such as time spent in
hypoxia, temperature, and humidity, and the limited
statistical power due to small sample sizes, limit the con-
clusions that can be drawn from these findings. Study quality
Th
h Most of the studies have been carried out at 4,500 m (or
equivalent), but Naughton et al.'s study [20] performed
at 1,829 and 2,438 m was unable to find any significant
differences between HH and NH. These altitudes corres-
pond to PO2 values of 118 mmHg (15.7 kPa) and
108 mmHg (14.4 kPa) [25] respectively, and it is possible
that these altitudes were not high enough to elicit differ-
ences in the measured variables. Significant differences
between the effects of NH and HH may impact the in-
terpretation and application of results from studies at
high altitude where the change in pressure may be a
confounding influence in the evaluation of physiological
responses to high altitude. Strengths and limitations of this study Although this is the first systematic review to summarise
crossover studies comparing physiological responses to
hypobaric and normobaric hypoxia, other publications
have come to similar conclusions on the topic. Millet
et al. [10] stress the importance of disentangling hypoxia
and hypobaria and Fulco et al. [26] highlight the need
for further investigations into NH versus HH, for par-
ticular application to pre-acclimatisation strategies. Standardisation of study methods and reporting may
aid interpretation of future studies and thereby improve
the quality of data in this area. This is important to im-
prove the quality of data that is used both for improving
understanding of hypoxia tolerance, both at altitude and
in the clinical setting. The strengths of this systematic review include the clear
research question, comprehensive search strategy and con-
sistent methods used for identifying eligible manuscripts
and extracting data. Limitations of this review include the
focus on crossover studies but are predominantly related
to the quantity and quality of the underlying literature. There are few studies that compare HH and NH and the
number of participants in each study is small. Whilst sev-
eral of these studies report interesting differences between
HH and NH, there is marked inconsistency in the reported
results. This may be due to a number of other factors
including heterogeneity of study design, duration and mag-
nitude of hypoxic dose and outcome reporting. Further-
more, the reporting of multiple phenotypes in each study
without correction for repeat testing may be associated
with an increased likelihood of type 1 errors. Conversely, ACTH: Adrenocorticotropic hormone; ADH: Anti-diuretic hormone;
AMS: Acute mountain sickness; ANP: Anti-naturetic protein; Bf: Respiratory
rate; BP: Blood pressure; CO2: Carbon dioxide; CO: Cardiac output;
ECG: Electrocardiogram; FetCO2: End tidal fraction of carbon dioxide;
FetO2: End tidal fraction of oxygen; exNO: Exhaled nitric oxide levels;
GFR: Glomerular filtration rate; GPX: Glutathione peroxidase; Hb
conc: Haemoglobin concentration; Hct: Haematocrit; HR: Heart rate;
LF/HF: Heart rate variability; pH2O: Humidity; HH: Hypobaric hypoxia;
HCR: Hypoxic cardiac response; HVR: Hypoxic ventilatory response; NO: Nitric
oxide; N2: Nitrogen; NH: Normobaric hypoxic; SpO2: Pulse oximetry;
PN2: Partial pressure of nitrogen; PO2: Partial pressure of oxygen;
PaCO2: Arterial partial pressures of carbon dioxide; PaO2: Arterial partial
pressures of oxygen; PetO2: End tidal partial pressure of oxygen; PETCO2: End
tidal partial pressure of CO2; Plasma K+: Plasma potassium concentration;
plasma Na+: Plasma sodium concentration; RQ: Respiratory quotient;
SaO2: Arterial blood saturations; SV: Stroke volume; urine Na+/K+: Urine
sodium-potassium ratio; VA: Alveolar ventilation; VCO2: Volume of CO2
produced; VE: Ventilation; VO2: Volume of oxygen consumed; VT: Tidal volume. Competing interests Competing interests 16. Roach RC, Loeppky JA, Icenogle MV. Acute mountain sickness: increased
severity during simulated altitude compared with normobaric hypoxia. J Appl Physiol. 1996;81:1908–10. The authors declare that they have no competing interests. 17. Savourey G, Launay JC, Besnard Y, Guinet A, Travers S. Normo- and
hypobaric hypoxia: are there any physiological differences? Eur J Appl
Physiol. 2003;89:122–6. Acknowledgements
We would like to thank Caroline De Brun. We would like to thank Caroline De Brun. 20. Naughton MT, Rochford PD, Pretto JJ, Pierce RJ, Cain NF, Irving LB. Is
normobaric simulation of hypobaric hypoxia accurate in chronic air-flow
limitation. Am J Resp Crit Care. 1995;152:1956–60. Received: 2 September 2014 Accepted: 18 December 2014 26. Fulco CS, Beidleman BA, Muza SR. Effectiveness of preacclimatization
strategies for high-altitude exposure. Exerc Sport Sci Rev. 2013;41:55–63. Abbreviations
ACTH Ad ACTH: Adrenocorticotropic hormone; ADH: Anti-diuretic hormone;
AMS: Acute mountain sickness; ANP: Anti-naturetic protein; Bf: Respiratory
rate; BP: Blood pressure; CO2: Carbon dioxide; CO: Cardiac output;
ECG: Electrocardiogram; FetCO2: End tidal fraction of carbon dioxide;
FetO2: End tidal fraction of oxygen; exNO: Exhaled nitric oxide levels;
GFR: Glomerular filtration rate; GPX: Glutathione peroxidase; Hb
conc: Haemoglobin concentration; Hct: Haematocrit; HR: Heart rate;
LF/HF: Heart rate variability; pH2O: Humidity; HH: Hypobaric hypoxia;
HCR: Hypoxic cardiac response; HVR: Hypoxic ventilatory response; NO: Nitric
oxide; N2: Nitrogen; NH: Normobaric hypoxic; SpO2: Pulse oximetry;
PN2: Partial pressure of nitrogen; PO2: Partial pressure of oxygen;
PaCO2: Arterial partial pressures of carbon dioxide; PaO2: Arterial partial
pressures of oxygen; PetO2: End tidal partial pressure of oxygen; PETCO2: End
tidal partial pressure of CO2; Plasma K+: Plasma potassium concentration;
plasma Na+: Plasma sodium concentration; RQ: Respiratory quotient;
SaO2: Arterial blood saturations; SV: Stroke volume; urine Na+/K+: Urine
sodium-potassium ratio; VA: Alveolar ventilation; VCO2: Volume of CO2
produced; VE: Ventilation; VO2: Volume of oxygen consumed; VT: Tidal volume. Page 20 of 20 Page 20 of 20 Coppel et al. Extreme Physiology & Medicine (2015) 4:2 Coppel et al. Extreme Physiology & Medicine (2015) 4:2 Coppel et al. Extreme Physiology & Medicine (2015) 4:2 Authors' contributions EGK, PH and MG conceived the idea of the review. JC, EGK and PH planned
the methodology and JC carried out the systematic review of the literature. The results were then checked by EGK and all authors discussed them. The
review was drafted by JC and then commented on by all authors. All authors
read and approved the final manuscript. 18. Savourey G, Launay JC, Besnard Y, Guinet-Lebreton A, Alonso A, Sauvet F,
et al. Normo or hypobaric hypoxic tests: propositions for the determination
of the individual susceptibility to altitude illnesses. Eur J Appl Physiol. 2007;100:193–205. 19. Hemmingsson T, Linnarsson D. Lower exhaled nitric oxide in hypobaric
than in normobaric acute hypoxia. Resp Physiol Neurobi. 2009;169:74–7. References 1. Grocott M, Montgomery H, Vercueil A. High-altitude physiology and
pathophysiology: implications and relevance for intensive care medicine. Crit Care. 2007;11:203. 2. Grant S, MacLeod N, Kay JW, Watt M, Patel S, Paterson A, et al. Sea level and
acute responses to hypoxia: do they predict physiological responses and
acute mountain sickness at altitude? Brit J Sport Med. 2002;36:141–6. 3. Millet GP, Faiss R, Brocherie F, Girard O. Hypoxic training and team sports:
a challenge to traditional methods? Brit J Sport Med. 2013;47:6–7. 4. DeHart RLE. Fundamentals of aerospace medicine. 2nd ed. Baltimore:
Williams and Wilkins; 1996. p. 90–1. 2. Grant S, MacLeod N, Kay JW, Watt M, Patel S, Paterson A, et al. Sea level and
acute responses to hypoxia: do they predict physiological responses and
acute mountain sickness at altitude? Brit J Sport Med. 2002;36:141–6. p
3. Millet GP, Faiss R, Brocherie F, Girard O. Hypoxic training and team sports:
a challenge to traditional methods? Brit J Sport Med. 2013;47:6–7. 4. DeHart RLE. Fundamentals of aerospace medicine. 2nd ed. Baltimore:
Williams and Wilkins; 1996. p. 90–1. 5. Conkin J, Wessel JH. Critique of the equivalent air altitude model. Aviat
Space Envir Md. 2008;79:975–82. 6. Self DA, Mandella JG, Prinzo OV, Forster EM, Shaffstall RM. Physiological
equivalence of normobaric and hypobaric exposures of humans to 25,000
feet (7620 m). Aviat Space Envir Md. 2011;82:97–103. 6. Self DA, Mandella JG, Prinzo OV, Forster EM, Shaffstall RM. Physiological
equivalence of normobaric and hypobaric exposures of humans to 25,000
feet (7620 m). Aviat Space Envir Md. 2011;82:97–103. p
7. Debevec T, Millet GP. Discerning normobaric and hypobaric hypoxia:
significance of exposure duration. J Appl Physiol. 2014;116:1255–5. 7. Debevec T, Millet GP. Discerning normobaric and hypobaric hypoxia:
significance of exposure duration. J Appl Physiol. 2014;116:1255–5. 8. Olivier G, Koehle MS, Guenette JA, Verges S, Chapman RF, Conkin J, et al. Comments on point: counterpoint: hypobaric hypoxia induces/does not
induce different responses from normobaric hypoxia. J Appl Physiol. 2012;112:1788–94. 8. Olivier G, Koehle MS, Guenette JA, Verges S, Chapman RF, Conkin J, et al. Comments on point: counterpoint: hypobaric hypoxia induces/does not
induce different responses from normobaric hypoxia. J Appl Physiol. 2012;112:1788–94. 9. Muza SR. Military applications of hypoxic training for high-altitude operations. Med Sci Sports Exerc. 2007;39:1625–31. 10. Millet GP, Faiss R, Pialoux V. Evidence for differences between hypobaric
and normobaric hypoxia is conclusive. Exerc Sport Sci Rev. 2013;41:133. Author details
1 21. Basualto-Alarcon C, Rodas G, et al. Cardiorespiratory parameters during
submaximal exercise under acute exposure to normobaric and hypobaric
hypoxia. Apunts Med Esport. 2012;47:65–72. 1University College London Centre for Altitude Space and Extreme
Environment Medicine, UCLH NIHR Biomedical Research Centre, Institute of
Sport and Exercise Health, 170 Tottenham Court Road, London W1T 7HA, UK. 2Integrative Physiology and Critical Illness Group, Clinical and Experimental
Sciences, Mailpoint 810, Sir Henry Wellcome Laboratories, Faculty of
Medicine, University of Southampton, University Hospital Southampton NHS
Foundation Trust, Tremona Road, Southampton SO16 6YD, UK. 3Anaesthesia
and Critical Care Research Unit, University Hospital Southampton NHS
Foundation Trust, Mailpoint 27, D Level, Centre Block, Tremona Road,
Southampton SO16 6YD, UK. 4NIHR Southampton Respiratory Biomedical
Research Unit, Southampton SO16 5ST, UK. 22. Tucker A, Reeves JT, Robertshaw D, Grover RF. Cardiopulmonary response to
acute altitude exposure—water loading and denitrogenation. Resp Physiol. 1983;54:363–80. 23. Degache F, Larghi G, Faiss R, Deriaz O, Millet G. Hypobaric versus
normobaric hypoxia: same effects on postural stability? High Alt Med Biol. 2012;13:40–5. 23. Degache F, Larghi G, Faiss R, Deriaz O, Millet G. Hypobaric versus
normobaric hypoxia: same effects on postural stability? High Alt Med Biol. 2012;13:40–5. 24. Isa KBM, Kawasaki N, Ueyama K, Sumii T, Kudo S. Effect of cold exposure
and shear stree on endothelial nitric oxide synthase activation. Biochem
Biophys Res Commun. 2011;412:318–22. 24. Isa KBM, Kawasaki N, Ueyama K, Sumii T, Kudo S. Effect of cold exposure
and shear stree on endothelial nitric oxide synthase activation. Biochem
Biophys Res Commun. 2011;412:318–22. 25. West JB, Milledge JS, Schoene RB, Luks A. High altitude medicine and
physiology. 5th ed. Boca Raton: CRC Press/Taylor & Francis Group; 2013. Received: 2 September 2014 Accepted: 18 December 2014 References Submit your next manuscript to BioMed Central
and take full advantage of:
• Convenient online submission
• Thorough peer review
• No space constraints or color figure charges
• Immediate publication on acceptance
• Inclusion in PubMed, CAS, Scopus and Google Scholar
• Research which is freely available for redistribution
Submit your manuscript at
www.biomedcentral.com/submit Submit your next manuscript to BioMed Central
and take full advantage of:
• Convenient online submission
• Thorough peer review
• No space constraints or color figure charges
• Immediate publication on acceptance
• Inclusion in PubMed, CAS, Scopus and Google Scholar
• Research which is freely available for redistribution
Submit your manuscript at
www.biomedcentral.com/submit 11. Conkin J. PH2O and simulated hypobaric hypoxia. Aviat Space Envir Md. 2011;82:1157–8. Submit your next manuscript to BioMed Central
and take full advantage of:
• Convenient online submission
• Thorough peer review
• No space constraints or color figure charges
• Immediate publication on acceptance
• Inclusion in PubMed, CAS, Scopus and Google Scholar
• Research which is freely available for redistribution
Submit your manuscript at
www.biomedcentral.com/submit Submit your next manuscript to BioMed Central
and take full advantage of: Submit your next manuscript to BioMed Central
and take full advantage of: 12. Faiss R, Pialoux V, Sartori C, Faes C, Deriaz O, Millet GP. Ventilation, oxidative
stress, and nitric oxide in hypobaric versus normobaric hypoxia. Med Sci
Sports Exerc. 2013;45:253–60. • Convenient online submission 13. Loeppky JA, Icenogle M, Scotto P, Robergs R, HinghoferSzalkay H, Roach RC. Ventilation during simulated altitude, normobaric hypoxia and normoxic
hypobaria. Resp Physiol. 1997;107:231–9. • Thorough peer review 14. Loeppky JA, Roach RC, Maes D, Hinghofer-Szalkay H, Roessler A, Gates L,
et al. Role of hypobaria in fluid balance response to hypoxia. High Alt Med
Biol. 2005;6:60–71. 15. Miyagawa K, Kamijo Y, Ikegawa S, Goto M, Nose H. Reduced hyperthermia-
induced cutaneous vasodilation and enhanced exercise-induced plasma
water loss at simulated high altitude (3,200 m) in humans. J Appl Physiol. 2011;110:157–65. 15. Miyagawa K, Kamijo Y, Ikegawa S, Goto M, Nose H. Reduced hyperthermia-
induced cutaneous vasodilation and enhanced exercise-induced plasma
water loss at simulated high altitude (3,200 m) in humans. J Appl Physiol. 2011;110:157–65.
|
https://openalex.org/W2781586396
|
https://jurnal.ugm.ac.id/jpti/article/download/25748/18824
|
English
| null |
Morphological and Molecular Observation to Confirm the Taxonomic of Coptocercus biguttatus (Coleoptera: Cerambycidae) on Cloves in Ambon and Part of Ceram Island
|
Jurnal perlindungan tanaman Indonesia/Jurnal Perlindungan Tanaman Indonesia
| 2,018
|
cc-by-sa
| 5,540
|
Morphological and Molecular Observation
to Confirm the Taxonomic of Coptocercus biguttatus (Coleoptera: Cerambycidae)
on Cloves in Ambon and Part of Ceram Island
Observasi Morfologi dan Molekuler
untuk Konfirmasi Taksonomi Coptocercus biguttatus (Coleoptera: Cerambycidae)
pada Tanaman Cengkih di Pulau Ambon dan Sebagian Pulau Seram
Mohamad Pamuji Setyolaksono1)*, Suputa2), & Nugroho Susetya Putra 2)
1)Regional Estate Crop Office on Seed Management and Plant Protection, Ambon
Jln. Pertanian-Passo, Ambon 97232
2)Department of Crop Protection, Faculty of Agriculture, Universitas Gadjah Mada,
Jln. Flora 1, Bulaksumur, Sleman, Yogyakarta 55281
*Corresponding author. E-mail: pamujislaksono@yahoo.com Morphological and Molecular Observation
to Confirm the Taxonomic of Coptocercus biguttatus (Coleoptera: Cerambycidae)
on Cloves in Ambon and Part of Ceram Island
Observasi Morfologi dan Molekuler
untuk Konfirmasi Taksonomi Coptocercus biguttatus (Coleoptera: Cerambycidae)
pada Tanaman Cengkih di Pulau Ambon dan Sebagian Pulau Seram
Mohamad Pamuji Setyolaksono1)*, Suputa2), & Nugroho Susetya Putra 2)
1)Regional Estate Crop Office on Seed Management and Plant Protection, Ambon
Jln. Pertanian-Passo, Ambon 97232
2)Department of Crop Protection, Faculty of Agriculture, Universitas Gadjah Mada,
Jln. Flora 1, Bulaksumur, Sleman, Yogyakarta 55281
*Corresponding author. E-mail: pamujislaksono@yahoo.com Morphological and Molecular Observation
to Confirm the Taxonomic of Coptocercus biguttatus (Coleoptera: Cerambycidae)
on Cloves in Ambon and Part of Ceram Island *Corresponding author. E-mail: pamujislaksono@yahoo.com Submitted June 7, 2017; accepted August 9, 2017 Submitted June 7, 2017; accepted August 9, 2017 INTISARI Penelitian ini dilakukan untuk mengonfirmasi spesies kumbang sungut panjang (Coptocercus biguttatus) yang
menggerek batang tanaman cengkih di Pulau Ambon dan sebagian dari Pulau Seram, Maluku menggunakan karakter
morfologi dan molekuler berbasis gen mtCO1 untuk sidik kekerabatan. Pengambilan sampel kumbang sungut panjang
dilakukan di Pulau Ambon dan sebagian dari Pulau Seram, Maluku dengan cara host rearing pada potongan batang
cengkih. Hasil penelitian menunjukkan bahwa C. biguttatus di Pulau Ambon dan sebagian dari Pulau Seram, Maluku
berkerabat dekat dan berada dalam satu kelompok dengan C. rubripes dan P. semipunctata di New Zealand dengan
nilai homologi 85%. Penyebaran C. biguttatus di Pulau Ambon dan sebagian dari Pulau Seram merata di seluruh
area pertanaman cengkih. Kata kunci: Cerambycidae, Coptocercus biguttatus, Maluku, penggerek batang cengkih ABSTRACT This research was conducted to confirm the species of longhorn beetle (Coptocercus biguttatus) drilling clove stems
in Ambon and part of Ceram Island, Moluccas, which has been noted as an important pest. Aim of this investigation was
to characterize the species morphologically, and more detailed with molecular technique via mtCO1 gene analysis. The
longhorn beetle was taken in Ambon and part of Ceram Island, Moluccas and then was etablished in laboratory with host
rearing method on pieces of clove stem. The results showed that C. biguttatus attacking clove stems in Ambon and part
of Ceram Island, Maluku was closely related and grouped into same cluster with C. rubripes and P. semipunctata in
of New Zealand with 85% homology value. C. biguttatus distributed evenly in all clove planting areas in Ambon and
part of Ceram Island. Keywords: Cerambycidae, clove stem borer, Coptocercus biguttatus, Mollucas Jurnal Perlindungan Tanaman Indonesia, Vol. 21, No. 2, 2017: 96–105
DOI: 10.22146/jpti.25748 Jurnal Perlindungan Tanaman Indonesia, Vol. 21, No. 2, 2017: 96–105
DOI: 10.22146/jpti.25748 Jurnal Perlindungan Tanaman Indonesia, Vol. 21, No. 2, 2017: 96–105
DOI: 10.22146/jpti.25748 Jurnal Perlindungan Tanaman Indonesia, Vol. 21, No. 2, 2017: 96–105
DOI: 10.22146/jpti.25748 INTRODUCTION damage living tissue, diseased, or died plants, which
were generally wooden plants (Amirullah et al., 2014). Clove belonged into to Family of Myrtaceace and
was one of Indonesia original spice crops coming from
Maluku Archipelago, in which the most of distributed
varieties in several places were Siputih, Sikotok,
Zanzibar, and Tuni (Pool et al., 1986). The insect pest
attacking clove stem in Ambon Island is believed as
stemborer (Boa, 1990), i.e. the longhorn beetle. However, the species of this pest has not been damage living tissue, diseased, or died plants, which
were generally wooden plants (Amirullah et al., 2014). Longhorn beetle (Coleoptera: Cerambycidae) has
been reported as pest on several agricultural com-
modities (Morillo et al., 2008). Putra et al. (2011)
stated that family of Cerambycidae was grouped
into family causing damage on cacao in Gianyar, Bali. In Central Sulawesi, cacao and coffee were reported
to be attacked by pest of Cerambycidae family
(Fahri & Sataral, 2015). Larvae of Cerambycidae Clove belonged into to Family of Myrtaceace and
was one of Indonesia original spice crops coming from
Maluku Archipelago, in which the most of distributed
varieties in several places were Siputih, Sikotok,
Zanzibar, and Tuni (Pool et al., 1986). The insect pest
attacking clove stem in Ambon Island is believed as
stemborer (Boa, 1990), i.e. the longhorn beetle. However, the species of this pest has not been Setyolaksono et al.: Morphological and Molecular Observation to Confirm the Taxonomic of Coptocercus biguttatus 97 clearly recognized and determined. This longhorned
beetle has been reported attacking the estate crops
like cacao (Kalshoven, 1981; Morillo et al., 2008) and
coffee (Rhainds et al., 2002), while the prevalence of
this insect on clove plants has not been reported yet. According to Kalshoven (1981), stemborers attacking
clove plants are determined as Nothopeus hemipterus,
N. fasciatipennis, and Hexamitodera semivelutina. Some previous explorations of longhorn beetle found
Subfamilies of Prioninae, Lepturinae, Cerambycinae,
and Lamiinae in National Park of Halimun Mountain
(Makihara et al., 2002); Subfamily of Lamiinae in
Bogor Great Garden (Noerdjito, 2010), in Mount of
Bromo regions (Makihara et al., 2011), in Lore
Lindu National Park (Fahri & Sataral, 2015), and in
forest areas of Walat Mountain (Sataral et al., 2015);
and Subfamilies of Lamiinae, Cerambycinae, and
Prioninae in Mount of Slamet regions (Noerdjito,
2011). Morphological Identification The characters of insect parts such as size, color,
pronotum, elytra, and genitalia were used in this study. Observation on genitalia was carried out by dissecting
abdominal apex to obtain genitalia using 0.5% of
NaCl solution. Whole observation was performed under
Microscope of Leica MZ16, and then was documented
with Optilab 2.1 application. INTRODUCTION gathered and transferred into absolute ethanol, and
then kept in refrigerator at temperature of -20oC for
molecular study as well as morphological study. MATERIALS AND METHODS The research was conducted from March 2016
until February 2017. The sampling of longhorn beetle
was conducted in clove planting areas and endemic
area of the stemborer at Ambon and part of Ceram
Island, Province of Maluku. Morphological identification
was done in Sub Laboratory of Pest Invertebrates,
while the molecular one was done in Sub Laboratory
of Virology, Department of Crop Protection, Faculty
of Agriculture, Universitas Gadjah Mada, Yogyakarta. DNA was amplified with universal primers of
forward LCO1490 (5’-GGTCAACAAATCATAAA
GATATTGG-3’) and reverse HCO2198 (5’-TAAA
CTTCAGGGTGACCAAAAAATCA-3’) amplifying
the fragment of mtCO1 gene at approximately 710 bp
in size (Folmer et al., 1994) using Automated Thermal
Cycler PTC-150 (MJ Research, Massachussets). The 25 µl of PCR reaction consisted of 0.5 µl KOD,
2.5 µl of Buffer, 2.5 µl of dNTPs, 1.5 µl of MgSO4,
16 µl of Nuclease-free water, 0.5 µl of forward
LCO1490 and reverse HCO2198 primers, as well Molecular Identification DNA extraction from insect parts such as head,
thorax, and fore leg (Rusterholz et al., 2015) was
conducted following the protocol of Goodwin et al. (1994). Those body parts were put into 1.5 ml eppendorf
tubes, added with 200 µl of CTAB (CTAB 2%,
NaCl 1.4 M, Tris-HCl 100 mM, EDTA 20 mM and
Polyvinylpyrrolidone (PVP) 1%, 2-mercaptoethanol),
and then grinded using micropastel. The extracted
solution was homogenized using vortex for 30 second
and was incubated at 65oC for 15 min in waterbath
(inversely mixing every 5 min). Afterwards, 200 µl
of Chloroform-Isoamyl Alcohol/CIAA (24:1) was
added and incubated at ambient temperature for
20−30 minutes prior to centrifugation at 8,000 rpm
for 5 minutes. The supernatant was transferred into
new 1.5 ml eppendorf tubes, and was followed by
adding 250 μl of cool absolute ethanol and 40 μl of
Natrium Asetat (NaOAc), and then was mixed gently. The mixed supernatant was incubated for 30 min at
temperature of -20oC and then was centrifuged at
11,500 rpm for 15 minutes. The supernatant was
removed and 200 μl of 70% cool ethanol was added
prior to centrifugation at 11,500 rpm for 15 minutes. The supernatant was then removed again and the
pellet was air-dried overnight, diluted into 50 µl of
TE 1× and homogenized by vortex. Study on longhorn beetle in Ambon Island has
not been previously reported, especially in relation
with cloves. This insect attacking cloves in Ambon
Island was assumed as exotic species due to the geo-
graphycal position of Ambon Island which is in
eastern side of Wallace line. This research was done
to characterize longhorn beetle which was endemic
insect on clove in Ambon Island, Maluku. Further-
more, a molecular characterization with mtCO1
gene was done to determine the relationship of this
beetle with other longhorned beetle in Ambon Island. Sampling Procedures The beetle was collected from the field by chopping
wood tissue showing the drilling hole at + 1.5 m
from the soil surface. The chopped tissues were then
incubated in closed room and covered with gauze
clothes to allow the adult of clove stemborers leave
the stems. Pupae obtained from field collection were
reared to become adults. The collected beetles were Jurnal Perlindungan Tanaman Indonesia Vol. 21 No. 2 98 as 1 µl of DNA template. Amplification was run
with pre-denaturation at 94oC for 2 min, 40 cycles
of denaturation at 98oC for 10 s, annealing at 55oC
for 20 s, extention at 68oC for 30 s, and final extention
at 68oC for 5 min. PCR product was visualized using
electrophoresis of 1% agarose gel and observed under
Bio-Rad UV Transilluminator 2000 after dipped
into 0.1% of ethidium bromide solution for 15 min. As much as 2 µl loading dye plus 5 µl of PCR product
was electrophorated at 50 Volt for 50 min with 1 kbp
DNA ladder. than others (Figure 1e). The length of antenna exceeded
the body length, with short smooth hair on whole
antenna, while the longer hair was appeared in the
lateral antenna which damped towards posterior. Spine was seen on 3rd to 7th segments of antenna. The 3rd segment of antenna was longer than scape of
4th and 5th segments (Figure 1a and 1b). There were five pronouncing tubercles on pronotum
in which two tubercles on anterior pronotum were
similar big circle; two bigger tubercles on posterior
pronotum were circle; and one oval tubercle which
was the biggest amongst others (Figure 1e). No
spine was arose on lateral pronotum. Pronotum was
characterized with very dense and smooth white
yellowish hair, excluded tubercle area on central
pronotum towards anterior near to head which was
distinguished by long disperse hair. The length of
male pronotum was 2.5−4.8 mm in range with
breadth of 1.9−3.5 mm in range; while the female
one was 2.7−4.7 mm in length and 1.9−3.2 mm in
breadth. Scutellum was semi-circle with similar
characteristic hair on pronotum (Figure 1e). Phylogenetic Analysis The samples were sent to the 1st Base DNA
Sequencing for DNA sequencing. Sequenced product
was aligned with Bioedit and ClustalOmega programs
to obtain consensus of base pair, and then matched
to the information of DNA sequence provided in
genebank. The percentage of homology for species
was generated from BLAST which was available in
NCBI (https://blast.ncbi.nlm.nih.gov/Blast.cgi). The
genetic diversity of alignment product was analyzed
with constructing phylogenetic tree through MEGA
7 program (Kumar et al., 2016) using method of
Maximum-Likelihood, Kimura 2 parameters and
1000 bootstrap. The phylogenetic tree was built by
comparing sampled insects to comparator insects
within same genus, and same tribe in sub family, as
well as inters sub family in family of Cerambycidae. The outgroup from NCBI was added to improve the
prediction of phylogenetic tree. Outgroup was selected
based on the closeness of relationship with analyzed
insects as well as their significant different with
other comparator insects (Dharmayanti, 2011). The dense, shoot, smooth and irregular punctures
on metasternum were also found (Figure 1f). Abdomen
consisting of 5 segments which was characterized by
smooth hair. Apical abdomen on male was indicated
with rounded-semi circle apex, while there was additional
truncated-like segment (not rounded-semi circle) on
female which was distinguished by dense smooth
hair (Figure 1g and 1h). Elytra was brown reddish, while head and pronotum
were darker than elytra. However, whole body of this
beetle was red brownish. Elytra was almost three
times longer than pronotum. The length of male elytra
was 5.2−13.1 mm in range, while female was about
5.7−13.4 mm in range. A pair of white spot closed to
suture was found on center of elytron in which its
size was getting bigger on lateral elytron. Meanwhile
such white yellowish spot was also generated on
basal of elytron and was getting bigger on lateral
elytron. White oval spot was characterized on apical
of elytron. A very dense puncture was appeared on
elytra in which the size was bigger than metasternum
and the position was deeper on basal to center of
elytra. However, puncture was getting sparse on
center of elytra to epical, the size was smaller and
the position was shallower. One hair was appeared
on each central puncture. The growing hair on
humerus region of puncture was curved and longer
than that on other region. Epical elytron was not Morphological Identification Morphological identification confirmed the species
as Coptocercus biguttatus. However, it had variations
in body length. For example, Hiroshi and Noerdjito
(2004) found that the body length of C. biguttatus
reached 19.1 mm. Anterior head was characterized
with short smooth hair and smaller than pronotum,
while lateral head did not bear such hair (Figure 1c). The site bearing antenna on head was pronouncing. No V or Y pattern was found on frons. Frons shape
was downward. Apical maxila palpi and labial palpi
broadened (Figure 1d). Type of antenna was philliform with 11 segments
in which the pedicel of second segment was smaller Setyolaksono et al.: Morphological and Molecular Observation to Confirm the Taxonomic of Coptocercus biguttatus 99 Figure 1. Adult of Coptocercus biguttatus; dorsal view (a), ventral view (b), Scape (sc), pedicel (pd), Antennomere III
(An III), Antennomere XI (An XI), lateral view (c), anterior view (d), Pronotum and antena (e), Metasternum (f),
scape (sc), pedicel (pd), Antennomere III (An III), scutellum (st), ventral view of male abdomen (g), ventral
view of female abdomen (h), abdominal sternite I (as I), abdominal sternite II (as II), abdominal sternite III
(as III), abdominal sternite IV (as IV), abdominal sternite V (as V), pygidium (py), Elytra (i), scutellum (st),
humerus (hu), pygidium (py), leg (j), Tarsal (k), Tarsus segment IV (l), male genitalia (m), female genitalia (n) Figure 1. Adult of Coptocercus biguttatus; dorsal view (a), ventral view (b), Scape (sc), pedicel (pd), Antennomere III
(An III), Antennomere XI (An XI), lateral view (c), anterior view (d), Pronotum and antena (e), Metasternum (f),
scape (sc), pedicel (pd), Antennomere III (An III), scutellum (st), ventral view of male abdomen (g), ventral
view of female abdomen (h), abdominal sternite I (as I), abdominal sternite II (as II), abdominal sternite III
(as III), abdominal sternite IV (as IV), abdominal sternite V (as V), pygidium (py), Elytra (i), scutellum (st),
humerus (hu), pygidium (py), leg (j), Tarsal (k), Tarsus segment IV (l), male genitalia (m), female genitalia (n) semi-cirle, but truncated-like and the generated spine
was very small and short (Figure 1i). could be visualized from several parts of their body
such as head, antenna, thorax, elytra, and abdomen. The legs were distinguished by sparse hair on
femur region, and dense hair on tibial region and
tarsus. Two identical long spurs were also found on
tibial region. Morphological Identification Formula of tarsus was 5-5-5 (Figure
1j), but it seemed to be 4-4-4, due to the 4th segment
of tarsus was hidden and the size was also shorter than
other segments (Figure 1k and 1l). The 3rd segment
was characterized by heavy peg. Wang (1998) elaborated
that the important characters for Cerambycidae,
Subfamily of Cerambycinae, tribe of Phorancanthini Molecular Identification Molecular identification of C. biguttatus was
performed in 4 stages, namely extraction of its DNA
total, amplification of mtCOI gene using universal
primers of LCO1490 (5’-GGTCAACAAATCATA
AAGATATTGG-3’) and HCO2198 (5’-TAAACTT
CAGGGTGACCAAAAAATCA-3’), electrophoresis
in order to recognize the quality of PCR product, and
sequencing to figure out the arrangement of nucleotide
in PCR product. The primers of LCO1490 and Jurnal Perlindungan Tanaman Indonesia 100 Vol. 21 No. 2 Vol. 21 No. 2 HCO2198 were universal which amplified the fragment
of mtCO1 gene at approximately 710 bp in size (Folmer
et al., 1994). This was complied with the visualization
of PCR product of mtCO1 gene amplification using
the above primers (Figure 2). The result of sequencing
found nucleotide of approximately 648 bp in size
(Figure 4). This was in line with Sharma and Kobayashi
(2014) who stated that the use of LCO1490 and
HCO2198 primers against various family, genus and
species could generate the variation in nucleotide
size of amplified mtCO1 gene fragment. The sequencing
was conducted to figure out the nucleotide arrangement
of PCR product in order to recognize the relationship
of sampled insect in built diagram of phylogenetic
tree. C. biguttatus was not listed yet in NCBI database,
so that its relationship was compared to Order, Sub-
family, Tribe, and Genus of similar insect from several
places in the world which had been recorded in
NCBI database. Sequence analysis was performed using Basic
Local Alignment Search Tool (BLAST) program on
National Center of Biotechnology Information (NCBI)
website. BLAST result indicated that C. biguttatus
which was found in Ambon and part of Ceram Island,
Maluku was identical with Coptocercus rubripes
(Accession Number in GenBank of KC593312) and
Phoracantha semipunctata (Accession Number in
GenBank of KC593339). That result was then utilized
to perform multiple alignment analysis with Bioedit
program and analysis of homology with MEGA 7
program to visualize the comparison between samples
and comparator. Phylogenetic analysis was demonstrated
using MEGA 7 program to build phylogenetic tree. The results of multiple alignment and homology
analyses expressed that samples of tested clove stem-
borer, C. biguttatus, was identical with wood borer beetles
from New Zealand i.e. C. rubripes and P. semipunctata. Sequences of those comparator insects were obtained
online from GenBank. Phylogenetic tree showed that C. biguttatus in
Ambon and part of Ceram Island, Maluku was closely
related and was grouped into same cluster with C. rubripesand P. Molecular Identification semipunctatain New Zealand (Figure 3)
with value of homology about 85% (Table 1). C. biguttatus was categorized into similar cluster to C. rubripes since these insects belonged into genus of
Coptocercus. Morphological features of both species
had some similarities such as length and color of body. However, they were different in some features. For example,
C. rubripes had two spines on apical elytra; the elytron
was characterized with yellow brownish lengthening
stripe on the central prior to basal; the lateral pronotum
possessed spherical tubercle; spine was also appeared
on lateral pronotum; and the body was red blackish so
that it was darker than C. biguttatus(Sopow et al., 2015). Meanwhile, it was closely related to P. semipunctata Figure 2. Electrophoresis of PCR product from amplification
of mtCO1 gene of Coptocercus biguttatus by
using primer of LCO1490-HCO2198 with 1kb
marker Figure 2. Electrophoresis of PCR product from amplification
of mtCO1 gene of Coptocercus biguttatus by
using primer of LCO1490-HCO2198 with 1kb
marker Figure 3. Phylogenetic tree of Coptocercus biguttatus (*) from Ambon and part of Ceram Island with comparator
species from GenBank database Figure 3. Phylogenetic tree of Coptocercus biguttatus (*) from Ambon and part of Ceram Island with comparator
species from GenBank database 101 Setyolaksono et al.: Morphological and Molecular Observation to Confirm the Taxonomic of Coptocercus biguttatus sumatranus was found in
Java and Sumatra; C. quatuordecimsignatus is found
in the Philippines; and C. q. celebensis is found on the
island of Sulawesi (Miroshnikov, 2016). since these insects were categorized into the same of
Tribe of Phoracanthini. Their morphological properties
were also similar. Some differences among them
were characterized by the elytra of P. semipunctata
which was dominated by brown with white yellow-
ish stripe on central region to suture. The stripe and
a pair of spines were found on apical elytra, and
spine was also generated on lateral pronotum (Wang,
1998; Sopow et al., 2015). C. biguttatus, C. rubripes,
and P. semipunctata were classified into same Sub-
family of Cerambycinae, the larger Subfamily in
Family of Cerambycidae. In addition, the tested
insect was also compared to insects of other Sub-
families such as Lamiinae and Spondylidinae, in
order to more completely figured out the relationship
with C. biguttatus. Comparator insects of Lamiinae
Subfamily were Apriona germari, Batocera rubus,
and Batocera horsfieldi from Tribe of Batocerini;
while those of Spondylidinae Subfamily were
Arhopalus syriacus, Tetropiumoreinum, and Tetropium
castaneum, which were belonged into Tribe of Asemini. The outgroup was added to achieve more trustable
information of sequence. Dharmayanti (2011) suggested
that outgroup should have related sequences with
samples and comparator with a few distinguisher. The
selected outgroup was Distenia undata of Family
Disteniidae, Subfamily Disteniinae, and Tribe Disteniini. This outgroup was chosen as it was also grouped
into same Superfamily with sampled and comparator
insects, i.e. Cerambycoidea. C. biguttatus was obtained from all host rearing
of clove stem. This fact indicated that C. biguttatus
was distributed evenly through all clove planting
areas in Ambon and part of Ceram Island (Figure 5). All collected samples found on main stem of clove
plants were belonged into Family of Myrtaceae. This was complied with Wang (1998) who stated that
longhorn beetle especially from Tribe of Phoracanthini
was associated with Families of Myrtaceae, Leguminosae
and Rutaceae, some conifers, genera Eucalyptus,
Acacia and citrus. Dry and wet seasons in Ambon
and part of Ceram Island did not influence the invasion
of C. biguttatus on clove plant. Investigation by
Dhahri et al. (2016) reported that infestation of
Phoracanthini Tribe (P. semipunctata and P. recurva)
on eucalyptus plant was not affected by season. Jurnal Perlindungan Tanaman Indonesia 102 Vol. 21 No. 2 Table 1. Percentage of homology from nucleotide base of Coptocercus biguttatusand comparator species from GenBank database
No. Species
1
2
3
4
5
6
7
8
9
10
1 Coptocercus biguttatus
ID
2 Coptocercus rubripes
85%
ID
3 Phoracantha semipunctata
85%
85%
ID
4 Batocera rubus
81%
78%
82%
ID
5 Batocera horsfieldi
80%
77%
77%
87%
ID
6 Apriona germari
78%
79%
81%
86%
85%
ID
7 Arhopalus syriacus
81%
79%
81%
83%
81%
82%
ID
8 Tetropium oreinum
77%
76%
78%
80%
79%
80%
81%
ID
9 Tetropium castaneum
76%
75%
77%
78%
79%
79%
81%
93%
ID
10 Distenia undata
45%
47%
51%
50%
48%
45%
45%
51%
49%
ID since these insects were categorized into the same of
Tribe of Phoracanthini. Their morphological properties
were also similar. Some differences among them
were characterized by the elytra of P. semipunctata
which was dominated by brown with white yellow-
ish stripe on central region to suture. The stripe and
a pair of spines were found on apical elytra, and
spine was also generated on lateral pronotum (Wang,
1998; Sopow et al., 2015). C. biguttatus, C. rubripes,
and P. semipunctata were classified into same Sub-
family of Cerambycinae, the larger Subfamily in
Family of Cerambycidae. In addition, the tested
insect was also compared to insects of other Sub-
families such as Lamiinae and Spondylidinae, in
order to more completely figured out the relationship
with C. biguttatus. Comparator insects of Lamiinae
Subfamily were Apriona germari, Batocera rubus,
and Batocera horsfieldi from Tribe of Batocerini;
while those of Spondylidinae Subfamily were
Arhopalus syriacus, Tetropiumoreinum, and Tetropium
castaneum, which were belonged into Tribe of Asemini. The outgroup was added to achieve more trustable
information of sequence. Dharmayanti (2011) suggested
that outgroup should have related sequences with
samples and comparator with a few distinguisher. The
selected outgroup was Distenia undata of Family
Disteniidae, Subfamily Disteniinae, and Tribe Disteniini. This outgroup was chosen as it was also grouped
into same Superfamily with sampled and comparator
insects, i.e. Cerambycoidea. biguttatus was also found on Ambon and part of
Ceram Islands, which was differed with previous
report by Gressitt (1951); Wang et al. (1996);
Brockerhoff & Bain (2000) who mentioned that this
species was only distributed in Papua New Guinea,
Australia, and New Zealand. In addition, other species
of Coptocercus such as C. Such condition was caused by the biological feature
of stemborer larvae which was active within clove
stem, so that it was not directly influenced by season. In their three reports, Hanks et al. (1991, 1993, and
1999) found that stemborers lived in plant stem by
penetrating the bark, destroying cambium and phloem
tissues for their survival, since the tissues contained
high nutrient. Larvae could stay in plant stem with
diameter of 5 cm. So, it is important for doing further
investigation to figure out the content of clove plants
that supports the survival of C. biguttatus. The outgroup was added to achieve more trustable
information of sequence. Dharmayanti (2011) suggested
that outgroup should have related sequences with
samples and comparator with a few distinguisher. The
selected outgroup was Distenia undata of Family
Disteniidae, Subfamily Disteniinae, and Tribe Disteniini. This outgroup was chosen as it was also grouped
into same Superfamily with sampled and comparator
insects, i.e. Cerambycoidea. Distribution of C. biguttatus This observation showed that invasion of C. biguttatus
on clove plant in Ambon and part of Ceram Island
was not affected by altitude as shown by fact that C. biguttatus could survive in host rearing system at low
places (15 m asl) as well as at high places (864 m asl)
(Table 2). This observation also confirmed that C. Setyolaksono et al.: Morphological and Molecular Observation to Confirm the Taxonomic of Coptocercus biguttatus 103 Regency
District
Village
Latitude
Altitude (m asl)
Kairatu
128,40284
-3,39683
44
Seruawan
128,44175
-3,42741
111
Western Seram
Kairatu
Kamarian
128,46828
-3,42839
406
Tihulale
128,49502
-3,43591
125
Rumahkay
128,53731
-3,40275
864
Central Maluku
Salahutu
Liang
128,33806
-3,50514
30
Waai
128,30879
-3,54769
155
Tulehu
128,30927
-3,57576
35
Suli
128,30625
-3,60143
62
West Leihitu
Hatu
128,05694
-3,68997
167
Liliboi
128,03759
-3,69836
539
Allang
128,01095
-3,73556
98
Wakasihu
127,98102
-3,74108
421
Leihitu
Seith
128,03487
-3,61492
266
Hila
128,10605
-3,58685
15
Mamala
128,21407
-3,54055
87
Morella
128,22626
-3,5194
110
Kota Ambon
Teluk Ambon Baguala
Passo
128,25577
-3,59784
255
Lateri
128,24606
-3,64102
44
Sirimau
Ema
128,24585
-3,70866
540
Nusaniwe
Amahusu
128,14882
-3,74103
427
Table 2. Distribution areas of Coptocercus biguttatus in Ambon and part of Ceram Island, Maluku
Figure 5. Distribution map of Coptocercus biguttatus on cloves planting areas in Ambon and part of Ceram Island, Maluku
Setyolaksono et al.: Morphological and Molecular Observation to Confirm the Taxonomic of Coptocercus biguttatus
103 Regency
District
Village
Latitude
Altitude (m asl)
Kairatu
128,40284
-3,39683
44
Seruawan
128,44175
-3,42741
111
Western Seram
Kairatu
Kamarian
128,46828
-3,42839
406
Tihulale
128,49502
-3,43591
125
Rumahkay
128,53731
-3,40275
864
Central Maluku
Salahutu
Liang
128,33806
-3,50514
30
Waai
128,30879
-3,54769
155
Tulehu
128,30927
-3,57576
35
Suli
128,30625
-3,60143
62
West Leihitu
Hatu
128,05694
-3,68997
167
Liliboi
128,03759
-3,69836
539
Allang
128,01095
-3,73556
98
Wakasihu
127,98102
-3,74108
421
Leihitu
Seith
128,03487
-3,61492
266
Hila
128,10605
-3,58685
15
Mamala
128,21407
-3,54055
87
Morella
128,22626
-3,5194
110
Kota Ambon
Teluk Ambon Baguala
Passo
128,25577
-3,59784
255
Lateri
128,24606
-3,64102
44
Sirimau
Ema
128,24585
-3,70866
540
Nusaniwe
Amahusu
128,14882
-3,74103
427
Table 2. Distribution areas of Coptocercus biguttatus in Ambon and part of Ceram Island, Maluku Figure 5. Distribution map of Coptocercus biguttatus on cloves planting areas in Ambon and part of Ceram Island, Maluku Figure 5. Distribution map of Coptocercus biguttatus on cloves planting areas in Ambon and part of Ceram Is LITERATURE CITED Hanks, L.M., T.D. Paine, & J.G. Millar. 1993. Host
Species Preference and Larval Performance in
the Wood-Boring Beetle Phoracantha semipunctata
F. Oecologia 95: 22−29. Amirullah, C. Ariani, & Suriana. 2014. Keaneka-
ragaman Kumbang Cerambycidae (Coleoptera)
di Kawasan Gunung Mekongga Desa Tinukari
Kecamatan Wawo Kabupaten Kolaka Utara
Provinsi Sulawesi Tenggara [Diversity Cerambycidae
Beetles (Coleoptera) in Mekongga Mountain
Area Tinukari Village Wawo North Kolaka District
of Southeast Sulawesi Province]. Biowallacea 1:
16−24. Hanks, L.M., T.D. Paine, J.G. Millar, C.D. Campbell,
& U.K. Schuch. 1999. Water Relations of Host Trees
and Resistance to the Phloem-Boring Beetle
Phoracantha semipunctata F. (Coleoptera:
Cerambycidae). Oecologia 119: 400−407. Hiroshi, M., & W.A. Noerdjito. 2004. Longicorn
Beetles of Museum Zoologicum Bogoriense,
Identified by Dr. E.F. Gilmour, 1963 (Coleoptera:
Disteniidae and Cerambycidae). Bulletin of FFPRI
3: 49-98. Boa, E. 1990. Clove Disease Project: Summary of
Research 1975−1990. Publications by the Clove
Diseases Project (ATA-71) and Associated Scientists. Balittro, Bogor. 36 p. Kalshoven L.G.E. 1981. Pest of Crops in Indonesia. (Revised and Translated by PA Van Der Laan). PT. Ichtiar Baru, Jakarta. 701 p. Brockerhoff, G.E. & J. Bain. 2000. Biosecurity
Implications of Exotic Beetles Attacking Trees
and Shrubs In New Zealand. New Zealand Plant
Protection 53: 321−327. Kumar, S., G. Stecher, & K. Tamura. 2016. MEGA7:
Molecular Evolutionary Genetics Analysis Version
7.0. Journal Molecular Biology and Evolution
33: 1870−1874. Dhahri S., F. Lieutier, F.C. Cheikhrouha, & M.L. Ben
Jamaa. 2016. Distribution, Preference and Per-
formance of Phoracantha recurva and Phoracantha
semipunctata (Coleoptera Cerambycidae) on
Various Eucalyptus Species in Tunisia. REDIA
99: 83−95. Makihara H., Sugiharto, & W.A. Noerdjito. 2011. Longicorn Beetles from Mts. Bromo in East
Java, Indonesia, with Descriptions of a New
Species and a New Subspecies (Coleoptera;
Cerambycidae). Bulletin of FFPRI 10: 251−261. Dharmayanti, N.L.P.I. 2011. Filogenetika Molekuler:
Metode Taksonomi Organisme Berdasarkan Sejarah
Evolusi [Molecular Phylogenetic: Organism
Taxonomy Method Based on Evolutionary History]. Wartazoa 21: 1−10. Makihara H., W.A. Noerdjito, & Sugiharto. 2002. Longicorn Beetles from Gunung Halimun National
Park, West Java, Indonesia from 1997−2002
(Coleoptera; Disteniidae and Cerambycidae). Bulletin of FFPRI 1: 189−223. Fahri & M. Sataral. 2015. Longhorn Beetle (Coleoptera:
Cerambycidae) in Enclave Area, Lore Lindu
National Park, Central Sulawesi. Online Journal
of Natural Science 4: 149−157. Miroshnikov, A.I. 2016. New Data on the Taxonomy
and Geographic Distribution of Oriental Species
of the Genus Coptocercus Hope, 1841 (Coleoptera:
Cerambycidae: Phoracanthini). Russian Entomological
Journal 25: 387−392. Folmer, O., M. Black, W. Hoeh, R. Lutz, & R. CONCLUSION Both morphological and molecular identification
are confirmed that C. biguttatus in Ambon and part
of Ceram Island, Maluku was closely related and
belonged into the same cluster with C. rubripes and
P. semipunctata in New Zealand. The author would like to thank to Argawi Kandito
for DNA extraction assistance and phylogeny discussion. The grateful expression was also delivered to BPPSDM
Ministry of Agriculture, Republic of Indonesia for
financial support and Regional Estate Crop Office
on Seed Management and Plant Protection of
Ambon for facilitating this postgraduate study. Jurnal Perlindungan Tanaman Indonesia 104 Vol. 21 No. 2 LITERATURE CITED Vrijenhoek. 1994. DNA Primers for Amplification of Mito-
chondrial Cytochrome C Oxidase Subunit I from
Diverse Metazoan Invertebrates. Molecular Marine
Biology and Biotechnology 3: 294−299. Morillo, F., P. Sanchez, C. Giron, A. Valera, W. Munoz,
& J. Guerra. 2008. Behavior of Cacao Hybrids
(Theobroma cacao) to Attack by Steirastoma breve
(Coleoptera: Cerambycidae). Revista Colombiana
de Entomologia 34: 151−155. Goodwin, P.H., B.G. Xue, C.R. Kuske, & M.K. Sears. 1994. Amplification of Plasmid DNA to Detect
Plant Pathogenic Mycoplasmalike Organisms. Annals of Applied Biology 124: 27−36. Noerdjito, W.A. 2010. Arti Kebun Raya Bogor bagi
Kehidupan Kumbang Sungut Panjang (Coleoptera,
Cerambycidae) [Meaning of the Bogor Botanical
Garden for Life Longhorn Beetles (Coleoptera,
Cerambycidae)]. Jurnal Biologi Indonesia 6:
289−292. Gressitt, J.L. 1951. Longicorn Beetles from New Guinea
and the South Pacific (Coleoptera: Cerambycidae). Annals of the Entomological Society of America
44: 1−30. Hanks, L.M., T.D. Paine, & J.G. Millar. 1991. Mechanisms of Resistance in Eucalyptus against
Larvae of the Eucalyptus Longhorned Borer
(Coleoptera: Cerambycidae). Entomological Society
of America 20: 1583−1588. Noerdjito, W.A. 2011. Evaluasi Kondisi Hutan
Berdasarkan Keragaman Kumbang Sungut Panjang
(Coleoptera; Cerambycidae) di Kawasan Gunung
Slamet [Evaluation of Forest Condition Based
on Diversity of Longhorn Beetle (Coleoptera;
Cerambycidae) in Slamet Mountain Area]. Berita Biologi 10: 521−531. 105 Setyolaksono et al.: Morphological and Molecular Observation to Confirm the Taxonomic of Coptocercus biguttatus Sataral, M., T. Atmowidi, & W.A. Noerdjito. 2015. Diversity and Abundance of Longhorn Beetles
(Coleoptera: Cerambycidae) in Gunung Walat
Educational Forest, West Java, Indonesia. Journal
of Insect Biodiversity 3: 1−12. Pool, P.A., S.J. Eden-Green, & M.T. Muhammad. 1986. Variation in Clove (Syzygium aromaticum)
Germplasm in the Moluccan Islands. Euphytica
35: 149−159. Putra, I.G.A.P., N.L. Watiniasih, & N.M. Suartini. 2011. Inventarisasi Serangga pada Perkebunan
Kakao (Theobroma cacao) Laboratorium Unit
Perlindungan Tanaman Desa Bedulu, Kecamatan
Blahbatuh, Kabupaten Gianyar, Bali [Inventory
of Insects on Cocoa Plantation (Theobroma cacao)
Laboratory of Plant Protection Unit, Bedulu
Village, Blahbatuh District, Gianyar Regency,
Bali]. Jurnal Biologi 14: 19−24. Sharma, P., & T. Kobayashi. 2014. Are “Universal”
DNA Primers Really Universal?. Journal of Applied
Genetics-Springer 55: 485−497. Sopow, S., B. Gresham, & J. Bain. 2015. Exotic
Longhorn Beetles (Coleoptera: Cerambycidae)
Established in New Zealand. New Zealand
Entomologist 38: 107−125. Wang, Q. 1998. The Phoracanthine Beetles (Coleoptera:
Cerambycidae: Cerambycinae): Taxonomic Overview
and Generic Relationships. Invertebrate Taxonomy
12: 667−684. Rhainds, M., L. Chinchiew, Z. Moli, & G. Gries. 2002. LITERATURE CITED Incidence, Symptoms, and Intensity of Damage
by Three Coffee Stemborers (Coleoptera:
Cerambycidae) in South Yunan, China. Journal
of Economic Entomology 95: 106−112. Wang, Q., I.W.B. Spineton, & T.R. New. 1996. Bio-
geography of the Phoracanthine Beetles (Coleoptera:
Cerambycidae). Journal of Biogeography 23:
75−94. Rusterholz, H.P., S. Ursenbacher, A. Coray, U. Weibel,
& B. Baur. 2015. DNA Quantity and Quality in
Remnants of Traffic-Killed Specimens of an
Endangered Longhorn Beetle: A Comparison of
Different Methods. Journal of Insect Science 15:
120−125.
|
https://openalex.org/W2885700920
|
https://nottingham-repository.worktribe.com/file/947692/1/The%20Reporting%20of%20IRAS%20Compliance.pdf
|
English
| null |
The Reporting on ERAS Compliance, Outcomes, and Elements Research (RECOvER) Checklist: A Joint Statement by the ERAS<sup>®</sup> and ERAS<sup>®</sup> USA Societies
|
World journal of surgery
| 2,018
|
cc-by
| 5,347
|
The Reporting on ERAS Compliance, Outcomes, and
Research (RECOvER) Checklist: A Joint Statement
by the ERAS and ERAS USA Societies Kevin M. Elias1 • Alexander B. Stone2 • Katharine McGinigle3 • Jo’An I. Tankou1 •
Michael J. Scott4,5 • William J. Fawcett6 • Nicolas Demartines7 • Dileep N. Lobo8 •
Olle Ljungqvist9 • Richard D. Urman2 on behalf of the ERAS Society and ERAS USA Kevin M. Elias1 • Alexander B. Stone2 • Katharine McGinigle3 • Jo’An I. Tankou1 •
Michael J. Scott4,5 • William J. Fawcett6 • Nicolas Demartines7 • Dileep N. Lobo8 •
Olle Ljungqvist9 • Richard D. Urman2 on behalf of the ERAS Society and ERAS USA The Author(s) 2018 & Kevin M. Elias
kelias@bwh.harvard.edu World J Surg
https://doi.org/10.1007/s00268-018-4753-0 ORIGINAL SCIENTIFIC REPORT Abstract Abstract
Background Enhanced recovery after surgery (ERAS) programs are multimodal care pathways designed to minimize
the physiological and psychological impact of surgery for patients. Increased compliance with ERAS guidelines is
associated with improved patient outcomes across surgical types. As ERAS programs have proliferated, an unin-
tentional effect has been significant variation in how ERAS-related studies are reported in the literature. Methods To improve the quality of ERAS reporting, ERAS USA and the ERAS Society launched an effort to bst act
Background Enhanced recovery after surgery (ERAS) programs are multimodal care pathways designed to minimize
the physiological and psychological impact of surgery for patients. Increased compliance with ERAS guidelines is
associated with improved patient outcomes across surgical types. As ERAS programs have proliferated, an unin-
tentional effect has been significant variation in how ERAS-related studies are reported in the literature. Methods To improve the quality of ERAS reporting, ERAS USA and the ERAS Society launched an effort to
create an instrument to assist authors in manuscript preparation. Criteria to include were selected by a combination of
literature review and expert opinion. The final checklist was refined by group consensus. tentional effect has been significant variation in how ERAS related studies are reported in the literature. Methods To improve the quality of ERAS reporting, ERAS USA and the ERAS Society launched an effort to
create an instrument to assist authors in manuscript preparation. Criteria to include were selected by a combination of
literature review and expert opinion. The final checklist was refined by group consensus. Results The Societies present the Reporting on ERAS Compliance, Outcomes, and Elements Research (RECOvER)
Checklist. The tool contains 20 items including best practices for reporting clinical pathways, compliance auditing,
and formatting guidelines. Results The Societies present the Reporting on ERAS Compliance, Outcomes, and Elements Research (RECOvER)
Checklist. The tool contains 20 items including best practices for reporting clinical pathways, compliance auditing,
and formatting guidelines. Conclusions The RECOvER Checklist is intended to provide a standardized framework for the reporting of ERAS-
related studies. The checklist can also assist reviewers in evaluating the quality of ERAS-related manuscripts. Authors are encouraged to include the RECOvER Checklist when submitting ERAS-related studies to peer-reviewed
journals. RECOvER development A checklist and statement were developed by a small
working group of volunteers from ERAS USA (the
American chapter of the ERAS Society). A subcommittee
(KME, KM, JIT) from the ERAS USA Research Com-
mittee reviewed ERAS-related publications from across
different medical specialties and study designs. In devel-
oping the checklist, subcommittee members were asked to
review 10–15 manuscripts each from anesthesia, surgery,
or general interest journals and tasked to define best
practices in ERAS reporting. Questions to be addressed
were: In recent years, research in ERAS has expanded sig-
nificantly. ERAS programs have expanded beyond col-
orectal surgery to other surgical disciplines and have been
implemented successfully in pancreatic surgery, thoracic
surgery, liver resection, urologic surgery, gynecologic
surgery, and emergency surgery, among others [5, 9–13]. An unintentional effect of this rapid expansion has been
significant variations in how ERAS studies are reported
[14]. The COMPAC (Core Outcome Measures in Periop-
erative and Anaesthetic Care) group has embarked on an
effort to standardize the outcomes reported in perioperative
medicine https://www.niaa-hsrc.org.uk/HSRC-COMPAC. While there are some efforts to apply a similarly tiered
approach to reporting ERAS outcomes, we believe that a
truly comprehensive ERAS report should detail not only
the outcomes, but also the process by which those out-
comes were achieved [15]. Like any other scientific
enterprise, the methods should contain sufficient detail to
enable another group to reproduce the results. Moreover, as
ERAS protocols have now been in place at many sites for
years, there is a need to mature ERAS studies beyond the
common retrospective comparisons to pre-ERAS historic
controls. These lower-quality studies tend to magnify the
benefits of the intervention being studied. Indeed, properly
designed prospective studies have been revealing, showing
that interventions which may improve outcomes under
traditional perioperative management do not necessarily
confer additional benefit when both groups are on ERAS
pathways [16]. Hence, there is a need to formalize ERAS-
related research such that meaningful results are repro-
ducible and generalizable. We created the Reporting on
ERAS Compliance, Outcomes, and Elements Research
(RECOvER) Checklist to provide authors and reviewers
with a set of standards for excellence in reporting ERAS- 1. How are the study groups defined? 2. How do the authors convey the implementation of
ERAS principles? 3. What steps are taken to assess compliance? 4. What outcomes are measured? Abstract 6
Department of Anaesthesia, Royal Surrey County Hospital
and University of Surrey, Guilford, UK
7
Department of Visceral Surgery, Lausanne University
Hospital CHUV, Lausanne, Switzerland
8
Gastrointestinal Surgery, Nottingham Digestive Diseases
Centre and National Institute for Health Research (NIHR)
Nottingham Biomedical Research Centre, Nottingham
University Hospitals NHS Trust and University of
Nottingham, Queen’s Medical Centre,
Nottingham NG7 2UH, UK
9
Department of Surgery, Faculty of Medicine and Health,
School of Health and Medical Sciences, O¨ rebro University,
O¨ rebro, Sweden 6
Department of Anaesthesia, Royal Surrey County Hospital
and University of Surrey, Guilford, UK kelias@bwh.harvard.edu
1
Division of Gynecologic Oncology, Department of
Obstetrics, Gynecology and Reproductive Biology, Brigham
and Women’s Hospital, Harvard Medical School, 75 Francis
Street, Boston, MA 02115, USA
2
Department of Anesthesiology, Perioperative and Pain
Medicine, Brigham and Women’s Hospital, Harvard Medical
School, Boston, MA, USA
3
Division of Vascular Surgery, University of North Carolina
School of Medicine, Chapel Hill, NC, USA
4
Department of Anesthesiology, Virginia Commonwealth
University Health System, Richmond, VA, USA
5
Department of Anesthesiology, Perelman School of
Medicine, University of Pennsylvania, Philadelphia, PA,
USA 7
Department of Visceral Surgery, Lausanne University
Hospital CHUV, Lausanne, Switzerland 1
Division of Gynecologic Oncology, Department of
Obstetrics, Gynecology and Reproductive Biology, Brigham
and Women’s Hospital, Harvard Medical School, 75 Francis
Street, Boston, MA 02115, USA 2
Department of Anesthesiology, Perioperative and Pain
Medicine, Brigham and Women’s Hospital, Harvard Medical
School, Boston, MA, USA 3
Division of Vascular Surgery, University of North Carolina
School of Medicine, Chapel Hill, NC, USA 4
Department of Anesthesiology, Virginia Commonwealth
University Health System, Richmond, VA, USA 4
Department of Anesthesiology, Virginia Commonwealth
University Health System, Richmond, VA, USA 5
Department of Anesthesiology, Perelman School of
Medicine, University of Pennsylvania, Philadelphia, PA,
USA 5
Department of Anesthesiology, Perelman School of
Medicine, University of Pennsylvania, Philadelphia, PA,
USA 12 12 123 World J Surg related studies. This checklist is not a guideline itself—in
fact, quite the opposite. It is a tool to assist authors when
reporting outcomes on guidelines already in practice and, if
anything, should prompt reviewers or readers to ask why
elements in a report are not described or may not be
appropriate. Our goal is to encourage reproducibility in
clinical studies, acknowledging the different variables
which may influence ERAS outcomes and the different
ways in which ERAS has been implemented in units
around the world. Abstract This is not a meta-analysis; rather, we
aim to improve study reporting so that future meta-analyses
can be more easily performed by ensuring sufficient
information
on
ERAS
practice
and
description
of
outcomes. Introduction Enhanced recovery after surgery (ERAS) programs are
multimodal care pathways designed to minimize the
physiological and psychological impact of surgery for
patients. ERAS pathways rely on multidisciplinary teams
and require coordinated interventions in all parts of peri-
operative care, from the initial preoperative consultation
through the hospitalization and onward to the return of the
patient to normal activities of daily living [1]. ERAS pro-
grams reduce hospital lengths of stay and postoperative
complications while decreasing the costs of care for
patients and health systems [2, 3]. Most ERAS pathways
are designed around approximately 25 core elements out-
lined by the ERAS Society [4]. There are data to suggest
that increased compliance with these core elements is
associated with improved outcomes across surgical types
[5–8]. RECOvER development This subcommittee developed an initial list of 32 items
for inclusion in a checklist of best practices. After dis-
cussion with the larger committee, this number was
reduced to 20 items to focus the checklist on elements
related to ERAS rather than to general guidelines for best
practices in research reporting. The total number of ele-
ments was reduced by removing those redundant with
general reporting guidelines, for example the Enhancing
QUAlity and Transparency Of health Research (EQUA-
TOR) network guidelines, or by combining similar ele-
ments
to
make
the
checklist
more
concise
[17]. Reconciliation in the rare cases where disagreement
occurred was achieved by following the majority opinion
of the authors. After agreement in the Research Committee
was achieved, the checklist was circulated to members of
the ERAS Society Executive Committee (MJS, WJF, ND,
DNL, and OLP) for further comments. Following feedback
from members of the society, the final number of 20 items 123 123 World J Surg Table 1 RECOvER Checklist for reporting of enhanced recovery research
Item
Recommendation Page Title
Title
1
Indicate that this is an enhanced recovery study in the title
Introduction
Background
2
Explain the area of uncertainty that the study seeks to address
Guidelines
3
If a published set of enhanced recovery guidelines exists for this procedure, include a reference to the
guidelines
Outcomes
4
Define the primary outcome and any key prespecified secondary outcomes for the study
Methods
IRB approval
5
Give the Institutional Review Board/Ethics Committee name and approval number. If permission was not
required, reasons should be stated
Study design
6
Indicate what type of study is presented (randomized controlled trial, cohort, cross-sectional, etc.) The
individual guidelines for the type of study should be followed (e.g., CONSORT for randomized
controlled trial, STROBE for cohort studies, etc.)
Setting
7
Describe whether this is a single or multicenter study, the type of practice (academic vs. community,
tertiary vs. RECOvER items The complete list of checklist items is shown in Table 1. We recommend that authors publishing research in the field
of ERAS include this checklist with their submissions and
indicate the location of each item in the manuscript. This is
a framework for guidance. It should facilitate publication
rather than serve as a barrier. The ultimate decision to
accept or reject manuscripts is with the individual journal
editors; the guidance is not proscriptive. Each item in the
checklist pertains to a particular point in the course of
conducting an ERAS research study, from conceptualiza-
tion to data analysis and the writing of the manuscript. Therefore, we recommend that the researchers consult the
checklist as early as possible during the study planning
process. Below we provide a detailed description of each
checklist item, followed by some examples. p
p
y
p
p
Within the materials and methods, all ERAS studies
should describe the Institutional Review Board (IRB) or
Ethics Committee review or explain the rationale for IRB/
Ethics Committee exemption (item 5). The study design,
including whether this is a prospective or retrospective
study, should be evident (item 6). The design description
should include details on the clinical context, including the
setting (item 7), timing (item 8), and selection of patients
(item 9) for the study. This includes the type of hospital,
period of recruitment, and inclusion and exclusion criteria
for the study. The authors should place the report tempo-
rally with respect to the introduction of ERAS at the
institution (item 10), including differentiating pre-ERAS
from post-ERAS groups of patients. An explicit statement
regarding the dates of introduction of ERAS at the insti-
tution, if possible, is preferred. Paramount to ERAS-related
studies is documentation that the principles of enhanced
recovery are being followed (item 11). While the literature
is rife with reports of ERAS failures, a closer inspection
may reveal a lack of compliance with ERAS concepts
[23, 24]. A detailed description of the ERAS pathway
should cover all phases of care (preadmission, preopera-
tive, intraoperative, post-anesthesia care, inpatient, post-
discharge, and follow-up care). The description should also
include the management strategies for perioperative opti-
mization, opioid-sparing analgesia, fluid management,
avoidance of starvation, nutritional care, mobilization, and
discharge. These elements must then be related to an audit
system for pathway compliance (item 12), whether the
ERAS Interactive Audit System (EIAS) or local
databases. RECOvER development primary), and the providers (limited group or all providers on a service)
Timing
8
Describe periods of recruitment, time points at which outcomes assessed, and follow-up
Participants
9
Define study inclusion and exclusion criteria
Enhanced recovery
protocol
10
Describe when the enhanced recovery protocol was implemented relative to the study period
11
Provide a flow diagram or table through the continuum of care detailing the enhanced recovery protocol
including the following elements:
(a) Preadmission patient education regarding the protocol
(b) Preadmission screening and optimization as indicated for nutritional deficiency, frailty, anemia,
HbA1c, tobacco cessation, and ethanol use
(c) Fasting and carbohydrate loading guidelines
(d) Preemptive analgesia (dose, route, timing)
(e) Anti-emetic prophylaxis (dose, route, timing)
(f) Intraoperative fluid management strategy
(g) Types, doses, and routes of anesthetics administered
(h) Patient warming strategy
(i) Management of postoperative fluids
(j) Postoperative analgesia and anti-emetic plans
(k) Plan for opioid minimization
(l) Drain and line management
(m) Early mobilization strategy
(n) Postoperative diet and bowel regimen management
(o) Criteria for discharge
(p) Tracking of post-discharge outcomes
Enhanced recovery
auditing
12
Describe the audit system for compliance with the enhanced recovery protocol and how compliance data
are measured
Outcomes
13
(a) Explain the criteria for assessing primary and secondary outcomes
(b) Distinguish among clinical, functional, administrative, and quality of life outcome measures
PROs
14
If patient questionnaires are used, provide references to validation of these study instruments
Results
Patient population
15
Use a flow diagram to explain the derivation of the study population
(a) Provide a Table I with the key demographic and clinical features of the study population
(b) Indicate number of participants with missing data for each variable of interest
Enhanced recovery
compliance
16
Provide a Table II with average compliance for each enhanced recovery protocol element and present a
comparison of the variation in enhanced recovery compliance among the study groups
Correlations
17
Perform logistic regression to correlate the change in primary outcome with the study intervention 12 3 World J Surg Table 1 continued
Item
Recommendation
Page
Discussion
Context
18
Explain what the study adds to the body of knowledge regarding the study intervention within the context
of enhanced recovery after surgery care
Limitations
19
Discuss the limitations of the study and how these might temper the findings
Other information
Funding
20
Document all sources of funding and potential conflicts of interest for the study authors
RECOvER Reporting on ERAS Compliance, Outcomes, and Elements Research, CONSORT Consolidated Standards Of Reporting Trials,
STROBE STrengthening the Reporting of OBservational studies in Epidemiology, PROs patient-reported outcomes Page was confirmed by consensus agreement by the members of
the Research Committee and Standards and Protocols
Committee of ERAS USA (ABS, RDU). RECOvER development length of stay, or clinical outcomes, such as wound infec-
tion or transfusion rates, there is considerable need for
more ERAS studies addressing functional outcomes. The
latter might include outcomes such as return to work or
discharge to home rather than to a rehabilitation facility
[18–20]. There is also a need for more studies examining
non-surgical perioperative morbidity within the context of
established ERAS programs, such as the consequences of
preoperative anxiety or postoperative delirium [21, 22]. RECOvER items The report should include a list of the metrics Reporting standards begin with the title page. ERAS
studies should refer to enhanced recovery within the title
(item 1), which will facilitate queries for future systematic
reviews. The title should also relay the study type and
surgical procedure studied—for example, a retrospective
cohort study of patients undergoing robotic-assisted pan-
creaticoduodenectomies. In the introduction, the authors
should explain the specific area of clinical uncertainty
being addressed within the context of ERAS (item 2)—for
instance, whether high-volume or high-concentration local
anesthetic provides superior local analgesia to the incision. As the ERAS Society and other perioperative research
societies have published guidelines for many procedures,
existing guidelines, if applicable, should be referenced
(item 3). The primary outcome for the study should be
clearly stated in the introduction, as well as key secondary
outcomes of interest (item 4). While many ERAS studies
have focused on administrative outcomes, such as hospital 123 Table 2 Example of a RECOvER Checklist
Item
Recommendation
Page
Title
Title
1
Gum chewing improves recovery of gut function within an enhanced recovery protocol for hepatic
resection
1
Introduction
Background
2
Whether gum chewing offers additional benefit for functional gut recovery after liver resection beyond
other enhanced recovery elements is uncertain
3
Guidelines
3
Melloul E, et al. World J Surg 2016 Oct;40(10):2425–2440
3
Outcomes
4
Primary outcome Time to first bowel movement after surgery
Secondary outcomes Incidence of postoperative ileus, length of stay, incidence of postoperative emesis
3
Methods
IRB approval
5
General Hospital IRB #123456
4
Study design
6
Retrospective cohort study
4
Setting
7
Single institution, community-based academic hospital with stable group of surgeons during the study
period
5
Timing
8
Patients included from March 2013–May 2015, events assessed daily from surgery to discharge, all
patients followed until 2-week postoperative visit
5
Participants
9
Inclusion criteria 18? RECOvER items and referred preoperatively for tobacco and ethanol counseling
(c) Fasting and carbohydrate loading guidelines
Normal diet until midnight, clear liquids until 2 h before surgery, 300-ml isotonic beverage containing a
total of 50 grams of maltodextrin finished 2 h before surgery
(d) Preemptive analgesia (dose, route, timing)
300 mg celecoxib, 500 mg acetaminophen both oral given in pre-op
(e) Anti-emetic prophylaxis (dose, route, timing)
4 mg ondansetron and 8 mg dexamethasone given intravenously prior to emergence
(f) Intraoperative fluid management strategy
Esophageal Doppler monitoring of stroke volume variation
(g) Types, doses, and routes of anesthetics administered
Continuous propofol, intravenous lidocaine, and low-dose ketamine infusion, no volatile anesthesia
(h) Patient warming strategy
Forced warm air and intravenous fluid warmer
(i) Management of postoperative fluids
0.5 ml/kg/h 9 6 h
(j) Postoperative analgesia and anti-emetic plans
0.25% liposomal bupivacaine wound infiltration, 500 mg acetaminophen and 600 mg ibuprofen every 6 h
orally, 4 mg ondansetron every 6 h intravenously as needed
World J Surg World J Surg 123 World J Surg Page Item
Recommendation
Page
(k) Plan for opioid minimization
First-line analgesic 25 mg tramadol every 6 h orally as needed, increased to 50 mg tramadol if needed,
followed by addition of IV lidocaine infusion if needed, followed by pregabalin 100–300 mg every 8 h
if needed, followed by 5–10 mg oral oxycodone for breakthrough pain
(l) Drain and line management
No routine wound drains, Foley catheter removed in OR
(m) Early mobilization strategy
Patients ambulate to chair in PACU, ambulate 9 3 starting postoperative day 0, out of bed all meals, out
of bed 8 h per day starting postoperative day 1
(n) Postoperative diet and bowel regimen management
Clear liquids post-op day 0, regular diet beginning post-op day 1, standing MiraLax daily beginning
post-op day 0
(o) Criteria for discharge
Tolerating at least 2000 ml po daily, voiding independently, pain well controlled on oral medication,
ambulating in hallways
(p) Tracking of post-discharge outcomes
Patients contacted by office through daily email survey
Enhanced recovery
auditing
12
All enhanced recovery elements charted by physician assistant into Enhanced Recovery Interactive Audit
System (EIAS)
8
Outcomes
13
(a) Primary outcome Bowel movement as documented by RN
Secondary outcomes Per patient report as collected by physician assistant interview
9
(b) Clinical outcomes
PROs
14
European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire-C30 (J Clin
Epidemiol 2014)
9
Results
Patient population
15
See Figure 1 (or similar)
10
(a) See Table 1 (or similar)
11
(b) Participants with missing data indicated in Table 1 footnotes
11
Enhanced recovery
compliance
16
Table II provides enhanced recovery compliance for the gum-chewing versus non-gum-chewing groups
for 15 metrics from the enhanced recovery pathway
12
Correlations
17
Table III provides logistic regression examining gum chewing with respect to primary and secondary
outcomes
13
Discussion
Context
18
Study suggests that gum chewing has additional benefits to standard bowel regimen, early feeding, and
laxative guidelines for promoting early return of gut function
15
Limitations
19
Not a prospective study, did not have sufficient power to subdivide patients by indication for hepatic
resection, poor compliance among the cohort with respect to early mobilization and termination of
intravenous fluids
16
Other information
Funding
20
Support from departmental grant
2
RECOvER Reporting on ERAS Compliance, Outcomes, and Elements Research, IRB Institutional Review Board, ICU intensive care unit,
NRS nutrition risk screening, PACU post-anesthesia care unit visualize the derivation and composition of the study
population (item 15). RECOvER items years old, participating in the enhanced recovery protocol, undergoing hepatic
resection, not admitted to ICU postoperatively
Exclusion criteria Age \18, unable or unwilling to participate in enhanced recovery protocol, other
surgical procedures, ICU admission
5
Enhanced recovery
protocol
10
enhanced recovery protocol was initiated in March 2012
6
11
Provide a flow diagram or table through the continuum of care detailing the enhanced recovery protocol
including the following elements:
7
(a) Preadmission patient education regarding the protocol
All patients receive an informational packet, watch a 10-minute video, and attend a 1-h preoperative
educational class
(b) Preadmission screening and optimization for nutritional deficiency, frailty, tobacco cessation, and
ethanol use
Patients are screened for nutritional deficiency using the NRS scoring system, frailty using the scoring
model published by Kim et al. RECOvER items A description of the population
should identify what proportion of all patients undergoing
the procedure of interest is being reported and the reasons
for exclusion from the study. The outcomes results should
be displayed in the context of the actual compliance with
the ERAS elements (item 16). Again, this requires an that enter into the compliance calculation. Similarly, out-
comes, both primary and secondary, should be clearly
defined a priori (item 13), and whenever patient-reported
outcomes (PROs) or surveys are introduced, these should
use validated and referenced instruments (item 14). Results reporting in ERAS should reflect similar trans-
parency to the methods. The reader should be able to 123 123 World J Surg provided commentary on the manuscript. Kevin Elias, MD, Gyne-
cologic Oncology/Critical Care Medicine, Brigham and Women’s
Hospital, Massachusetts, Committee Chairman. Maria Adamian,
MBA, CRNA, Health Administration, CarePoint Health, New Jersey. Frederic
Bushnell,
MD,
Anesthesiology,
Shriners
International
Hospitals for Children, Florida. Arman Dagal, MD, Anesthesiology,
University of Washington, Washington. Chad Engan, MD, General
Surgery, Great Falls Surgical Associates, Montana. Joseph Marcotte,
MD, General Surgery, Cooper Health, New Jersey. Katharine
McGinigle, MD, Vascular Surgery, University of North Carolina
School of Medicine, Chapel Hill, North Carolina. Nicollete Pauksta,
MSNA, BSN, CRNA, Anesthesiology, South Carolina. Venu Pil-
larisetty,
MD,
Hepatobiliary/Pancreatic
Surgery,
University
of
Washington,
Washington. Jo’An
Tankou, MD,
Obstetrics
and
Gynecology, Brigham and Women’s Hospital and Massachusetts
General Hospital, Massachusetts. provided commentary on the manuscript. Kevin Elias, MD, Gyne-
cologic Oncology/Critical Care Medicine, Brigham and Women’s
Hospital, Massachusetts, Committee Chairman. Maria Adamian,
MBA, CRNA, Health Administration, CarePoint Health, New Jersey. Frederic
Bushnell,
MD,
Anesthesiology,
Shriners
International
Hospitals for Children, Florida. Arman Dagal, MD, Anesthesiology,
University of Washington, Washington. Chad Engan, MD, General
Surgery, Great Falls Surgical Associates, Montana. Joseph Marcotte,
MD, General Surgery, Cooper Health, New Jersey. Katharine
McGinigle, MD, Vascular Surgery, University of North Carolina
School of Medicine, Chapel Hill, North Carolina. Nicollete Pauksta,
MSNA, BSN, CRNA, Anesthesiology, South Carolina. Venu Pil-
larisetty,
MD,
Hepatobiliary/Pancreatic
Surgery,
University
of
Washington,
Washington. Jo’An
Tankou, MD,
Obstetrics
and
Gynecology, Brigham and Women’s Hospital and Massachusetts
General Hospital, Massachusetts. auditing system in place so that percentage compliance
with the protocol can be plotted against the outcomes of
interest. Whenever possible, regression analysis techniques
should be used to test for independent associations between
the primary outcome and the intervention under study (item
17). References 1. Ljungqvist O, Scott M, Fearon KC (2017) Enhanced recovery
after surgery: a review. JAMA Surg 152:292–298 2. Lau CS, Chamberlain RS (2017) Enhanced recovery after surgery
programs improve patient outcomes and recovery: a meta-anal-
ysis. World J Surg 41:899–913. https://doi.org/10.1007/s00268-
016-3807-4 3. Visioni A, Shah R, Gabriel E et al (2018) Enhanced recovery
after surgery for noncolorectal surgery?: A systematic review and
meta-analysis of major abdominal surgery. Ann Surg 267:57–65 4. Elias KM (2017) Understanding enhanced recovery after surgery
guidelines: an introductory approach. J Laparoendosc Adv Surg
Tech A 27:871–875 Compliance with ethical standards Conflict of interest MS has received honoraria from Merck, Baxter,
Deltex and Cheetah Medical for lecturing and travel expenses. RU has
received research funding from Medtronic, Mallinckrodt, Merck, and
honorarium from 3M. DNL has received unrestricted research fund-
ing and speaker’s honoraria from Fresenius Kabi, BBraun and Baxter
Healthcare for unrelated work. OL has received honoraria from
Merck, Fresenius Kabi, Nutricia, Baxter and Abbot for advice or
lecturing and travel expenses. OL is a shareholder of Encare AB,
Sweden, an IT company involved in audit. WJF reports honoraria
from Merck, Baxter, Grunethal and Smiths for lecturing and travel
expenses. The remaining authors have no disclosures. Open Access This article is distributed under the terms of the
Creative Commons Attribution 4.0 International License (http://crea
tivecommons.org/licenses/by/4.0/), which permits unrestricted use,
distribution, and reproduction in any medium, provided you give
appropriate credit to the original author(s) and the source, provide a
link to the Creative Commons license, and indicate if changes were
made. RECOvER scope An example of the RECOvER Checklist appears in
Table 2. The primary aim of the RECOvER Checklist is to
ensure an ERAS-specific addendum accompanies ERAS-
related studies so that the reader can assess the ERAS-
specific elements. The checklist is not mandatory; rather, it
serves as a framework to make it easier to compare ERAS
studies and assemble future systematic reviews and meta-
analyses. RECOvER items For example, in a study of ambulation from the
postoperative recovery unit compared to ambulation on
reaching the inpatient ward, where the primary outcome is
actually the 6-minute walk test result at 2 weeks after
surgery, a regression analysis should include confounders
for early ambulation such as time of day, neuraxial anal-
gesia, or receipt of opioids. The discussion of ERAS studies should place the work
within the larger context of ERAS-related care (item 18). Authors should strive to link the findings with tangible
opportunities to improve clinical practice. A study that
shows a decrease in visual analog scale (VAS) pain scores
from 5 to 4 is of much less impact on the field than a
similar study that examines the proportion of patients dis-
charged to home rather than to a rehabilitation facility. Authors sometimes consciously or subconsciously overin-
terpret the results of their study, that is, they add ‘‘spin’’ to
the conclusions of a scientific report. Spin is defined as a
non-neutral way of reporting that distorts the interpretation
of results and misleads readers [25]. An appraisal of the
study limitations should be candid (item 19), including
critiques of the ERAS protocol itself, if indicated. Finally,
authors must be open regarding funding support and pos-
sible conflicts of interest (item 20). Acknowledgements The authors wish to thank the members of the
ERAS USA Research Committee who reviewed the checklist and RECOvER Checklist availability The RECOvER Checklist will be made available on the
ERAS USA as well as the ERAS Society websites. It is
free and available as an open access document to support
higher-quality and more consistent reporting of ERAS
research. 5. Gustafsson UO, Hausel J, Thorell A et al (2011) Adherence to the
enhanced recovery after surgery protocol and outcomes after
colorectal cancer surgery. Arch Surg 146:571–577 6. Gustafsson UO, Oppelstrup H, Thorell A et al (2016) Adherence
to the ERAS protocol is associated with 5-year survival after
colorectal cancer surgery: a retrospective cohort study. World J
Surg 40:1741–1747. https://doi.org/10.1007/s00268-016-3460-y 6. Gustafsson UO, Oppelstrup H, Thorell A et al (2016) Adherence
to the ERAS protocol is associated with 5-year survival after
colorectal cancer surgery: a retrospective cohort study. World J
Surg 40:1741–1747. https://doi.org/10.1007/s00268-016-3460-y g
p
g
y
7. Simpson JC, Moonesinghe SR, Grocott MP et al (2015)
Enhanced recovery from surgery in the UK: an audit of the g
p
g
y
7. Simpson JC, Moonesinghe SR, Grocott MP et al (2015)
Enhanced recovery from surgery in the UK: an audit of the 12 123 World J Surg enhanced recovery partnership programme 2009–2012. Br J
Anaesth 115:560–568 17. Moher D, Schulz KF, Simera I et al (2010) Guidance for devel-
opers of health research reporting guidelines. PLoS Med
7:e1000217 8. Group EC (2015) The impact of enhanced recovery protocol
compliance on elective colorectal cancer resection: results from
an international registry. Ann Surg 261:1153–1159 18. Callesen T, Klarskov B, Bech K et al (1999) Short convalescence
after inguinal herniorrhaphy with standardised recommendations:
duration and reasons for delayed return to work. Eur J Surg
165:236–241 9. Xiong J, Szatmary P, Huang W et al (2016) Enhanced recovery
after surgery program in patients undergoing pancreaticoduo-
denectomy: a PRISMA-compliant systematic review and meta-
analysis. Medicine (Baltimore) 95:e3497 19. Myles PS, Shulman MA, Heritier S et al (2017) Validation of
days at home as an outcome measure after surgery: a prospective
cohort study in Australia. BMJ Open 7:e015828 10. Huang H, Ma H, Chen S (2018) Enhanced recovery after surgery
using uniportal video-assisted thoracic surgery for lung cancer: a
preliminary study. Thorac Cancer 9:83–87 20. Fawcett WJ, Mythen MG, Scott MJ (2012) Enhanced recovery:
More
than
just
reducing
length
of
stay? Br
J
Anaesth
109:671–674 11. RECOvER Checklist availability Semerjian A, Milbar N, Kates M et al (2018) Hospital charges
and length of stay following radical cystectomy in the enhanced
recovery after surgery era. Urology 111:86–91 21. Petersen PB, Jorgensen CC, Kehlet H et al (2017) Delirium after
fast-track hip and knee arthroplasty—a cohort study of 6331
elderly patients. Acta Anaesthesiol Scand 61:767–772 y
g y
gy
12. Nelson G, Altman AD, Nick A et al (2016) Guidelines for
postoperative care in gynecologic/oncology surgery: enhanced
recovery after surgery (ERAS(R)) Society recommendations—
part II. Gynecol Oncol 140:323–332 y
22. Keller DS, Tantchou I, Flores-Gonzalez JR et al (2017) Predict-
ing delayed discharge in a multimodal enhanced recovery path-
way. Am J Surg 214:604–609 13. Paduraru M, Ponchietti L, Casas IM et al (2017) Enhanced
recovery after emergency surgery: a systematic review. Bull
Emerg Trauma 5:70–78 23. Dickson EL, Stockwell E, Geller MA et al (2017) Enhanced
recovery program and length of stay after laparotomy on a
gynecologic oncology service: a randomized controlled trial. Obstet Gynecol 129:355–362 g
14. Day RW, Fielder S, Calhoun J et al (2015) Incomplete reporting
of enhanced recovery elements and its impact on achieving
quality improvement. Br J Surg 102:1594–1602 24. Phan TD, D’Souza B, Rattray MJ et al (2014) A randomised
controlled trial of fluid restriction compared to oesophageal
Doppler-guided goal-directed fluid therapy in elective major
colorectal surgery within an enhanced recovery after surgery
program. Anaesth Intensive Care 42:752–760 15. Moonesinghe SR, Grocott MPW, Bennett-Guerrero E et al (2017)
American Society for Enhanced Recovery (ASER) and Periop-
erative Quality Initiative (POQI) joint consensus statement on
measurement to maintain and improve quality of enhanced
recovery pathways for elective colorectal surgery. Perioper Med
(Lond) 6:6 25. Boutron I, Dutton S, Ravaud P et al (2010) Reporting and
interpretation of randomized controlled trials with statistically
nonsignificant
results
for
primary
outcomes. JAMA
303:2058–2064 16. Rollins KE, Lobo DN (2016) Intraoperative goal-directed fluid
therapy in elective major abdominal surgery: a meta-analysis of
randomized controlled trials. Ann Surg 263:465–476 123 123
|
https://openalex.org/W4308279134
|
https://www.researchsquare.com/article/rs-2042808/latest.pdf
|
English
| null |
Transition practices for survivors of childhood cancer: A report from the Children's Oncology Group
|
Research Square (Research Square)
| 2,022
|
cc-by
| 6,881
|
Transition practices for survivors of childhood
cancer: A report from the Children's Oncology Group
Jordan Gilleland Marchak
(
jgillel@emory.edu
)
i
i
S h
l
f
di i
fl
C
d
l
d
i
d
C
Child
'
l h
f Cam Escoffery
Rollins School of Public Health, Emory University Transition practices for survivors of childhood
cancer: A report from the Children's Oncology Group
Jordan Gilleland Marchak
(
jgillel@emory.edu
)
Emory University School of Medicine; Aflac Cancer and Blood Disorders Center, Children's Healthcare of
Atlanta
Karim T. Sadak
University of Minnesota Masonic Children's Hospital and Cancer Center
Karen E. Effinger
Emory University School of Medicine; Aflac Cancer and Blood Disorders Center, Children's Healthcare of
Atlanta
Regine Haardörfer
Rollins School of Public Health, Emory University
Cam Escoffery
Rollins School of Public Health, Emory University
Karen Kinahan
Robert H. Lurie Comprehensive Cancer Center of Northwestern University
David R. Freyer
Children's Hospital of Los Angeles
Eric J. Chow
Fred Hutchinson Cancer Center, Seattle Children's Hospital
Ann Mertens
Emory University School of Medicine; Aflac Cancer and Blood Disorders Center, Children's Healthcare of
Atlanta
Research Article
Keywords: Transition to Adult Care, Late effects services, Pediatric cancer, Survivorship
Posted Date: November 4th, 2022
DOI: https://doi.org/10.21203/rs.3.rs-2042808/v1
License:
This work is licensed under a Creative Commons Attribution 4.0 International
License. Read Full License Results Representatives from 137 COG sites reported on institutional transition practices. Two-thirds (66.4%) of
sites discharge survivors to another institution for cancer-related follow-up care in adulthood. Transfer to
primary care (33.6%) was a commonly reported model of care for young adult-aged survivors. Sites
transfer at ≤ 18 years (8.0%), ≤ 21 years (13.1%), ≤ 25 years (7.3%), ≥ 26 years (12.4%), or when
survivors are "ready" (25.5%). Few institutions reported offering services aligned with the structured
transition process from the Six Core Elements (Median = 1, Mean = 1.56, SD = 1.54, range: 0–5). The most
prevalent barriers to transitioning survivors to adult care were perceived lack of late-effects knowledge
among clinicians (39.6%) and perceived lack of survivor desire to transfer care (31.9%). Purpose Pediatric healthcare systems must support childhood cancer survivors to optimize their transition to adult
care. This study aimed to assess the state of healthcare transition services provided by Children's
Oncology Group (COG) institutions. Conclusions Most COG institutions transfer adult-aged survivors of childhood cancer elsewhere for survivor care, yet
few programs report delivering recognized standards for quality healthcare transition programming to
support survivors. Methods A 190-question online survey was distributed to 209 COG institutions to assess survivor services,
including transition practices, barriers, and implementation of services aligned with the Six Core Elements
of Health Care Transition 2.0 from the US Center for Health Care Transition Improvement. Research Article Additional Declarations: No competing interests reported. Page 1/18 Version of Record: A version of this preprint was published at Journal of Cancer Survivorship on March
4th, 2023. See the published version at https://doi.org/10.1007/s11764-023-01351-y. Version of Record: A version of this preprint was published at Journal of Cancer Survivorship on March
4th, 2023. See the published version at https://doi.org/10.1007/s11764-023-01351-y. Version of Record: A version of this preprint was published at Journal of Cancer Survivorship on March
4th, 2023. See the published version at https://doi.org/10.1007/s11764-023-01351-y. Page 2/18 Introduction Historical healthcare transition practices in pediatric oncology have been insufficient, given that fewer
than half of adult-aged survivors of childhood cancer remain actively engaged in survivor care [9-11]. Transition to adult survivor care is a complicated process involving patient-, family-, provider-, and health
system-related factors [8, 12-14]. To improve transition outcomes, the US Center for Health Care
Transition (Got Transition®; https://www.gottransition.org/) developed and disseminated Six Core
Elements of Health Care Transition [15 16] These Six Core Elements offer a framework for clinicians and Elements of Health Care Transition [15, 16]. These Six Core Elements offer a framework for clinicians and
institutions to provide a structured transition process with patients and their caregivers, beginning early in
adolescence and continuing into young adulthood. The Six Core Elements of Health Care Transition have
been endorsed by the American Academy of Pediatrics (AAP), American Academy of Family Physicians
(AAFP), and the American College of Physicians (ACP) [15] and incorporated into consensus statements
on managing the transition to adult-centered healthcare for youth with a wide variety of special
healthcare needs [17-20]. Although healthcare transition research has been repeatedly called for in the pediatric oncology
literature [8, 12, 14, 21, 22], few data have been published characterizing transition support services for
survivors of childhood cancer [23, 24]. Future efforts to improve access and engagement in risk-based
care among adult-aged survivors must be informed by the gaps and barriers to healthcare transition
services offered by pediatric oncology institutions. The purpose of this study was to describe the state of
healthcare transition services provided by COG institutions, perceived barriers to transferring survivors to
adult long-term follow-up care, and implementation of health system services aligned with the Six Core
Elements of Health Care Transition. Methods We conducted a cross-sectional survey to assess survivorship services offered by COG institutions [25],
including healthcare transition supports. An online survey was made available from October 26, 2017, to
February 23, 2018, on SurveyMonkey. Emory University's Institutional Review Board (IRB) determined the
study to be exempt from IRB review. Introduction Approximately 85% of children and adolescents diagnosed with cancer will become long-term survivors
[1], leading to a growing survivor population with unique healthcare needs. Studies have shown that 60-
90% of adult-aged survivors of childhood cancer have ≥1 chronic condition, with 25-80% having severe or
life-threatening conditions [2, 3]. The National Academy of Medicine and Children's Oncology Group
(COG) recommend that survivors participate in continuous, risk-based survivor care across the lifespan to
monitor for late effects of their previous cancer treatments [4, 5]. Facilitating healthcare transition and
eventual transfer of risk-based survivor care from pediatric to adult settings is critical to the long-term Page 3/18 Page 3/18 health of childhood cancer survivors since their morbidity and mortality risks increase significantly over
time and do not reach a plateau [2, 3, 6]. Healthcare transition has been conceptualized as a process leading to an event: the process necessitates
gradual, planned movement toward an adult-centered model for healthcare (i.e., transition), while the
event occurs when the patient transfers from a pediatric to an adult healthcare team (i.e., transfer) [7, 8]. Historical healthcare transition practices in pediatric oncology have been insufficient, given that fewer
than half of adult-aged survivors of childhood cancer remain actively engaged in survivor care [9-11]. Transition to adult survivor care is a complicated process involving patient-, family-, provider-, and health
system-related factors [8, 12-14]. To improve transition outcomes, the US Center for Health Care
Transition (Got Transition®; https://www.gottransition.org/) developed and disseminated Six Core
Elements of Health Care Transition [15, 16]. These Six Core Elements offer a framework for clinicians and
institutions to provide a structured transition process with patients and their caregivers, beginning early in
adolescence and continuing into young adulthood. The Six Core Elements of Health Care Transition have
been endorsed by the American Academy of Pediatrics (AAP), American Academy of Family Physicians
(AAFP), and the American College of Physicians (ACP) [15] and incorporated into consensus statements
on managing the transition to adult-centered healthcare for youth with a wide variety of special
healthcare needs [17-20]. Healthcare transition has been conceptualized as a process leading to an event: the process necessitates
gradual, planned movement toward an adult-centered model for healthcare (i.e., transition), while the
event occurs when the patient transfers from a pediatric to an adult healthcare team (i.e., transfer) [7, 8]. Measures The current analysis is derived from a subset of a 190-item online survey assessing cancer survivorship
practices, services, and care delivery at COG institutions [25]. In follow-up to a prior survey of COG
survivorship practices [26], respondents were asked to report institutional characteristics, including
institution type, size, upper age limits for newly diagnosed patients, institutional policies on age at
transfer, and models of care for adult survivors of childhood cancer. If respondents reported their institution transferred patients out for long-term follow-up care in adulthood
(e.g., their institution transferred patients at a certain age or transferred survivors "when they are ready"),
they were asked to complete nine items related to healthcare transition programming. These items
included identifying 1) the top two most difficult barriers to transitioning survivors to adult care providers
for cancer-related care, 2) care team members involved in introducing and coordinating transition, 3)
when in the cancer trajectory sites typically introduce the concept of transition, and 4) if sites had
implemented transition programming (6 items) aligned with the Six Core Elements of Health Care
Transition 2.0 from the US Center for Health Care Transition Improvement (14). If respondents reported
their patients are "seen indefinitely and not transferred elsewhere," they were not asked any further
questions about institutional healthcare transition supports. All respondents were allowed to omit
responses to individual questions at their discretion. Sample & Procedures Page 4/18
Potential respondents were COG Principal Investigators (PIs) or the institutional contact in the COG Late
Effects Directory of Services at existing member institutions (n=209). COG leadership sent an
introductory email and survey instructions to one individual per institution. Following the introductory
email, Emory University researchers emailed a link to complete the online survey. Two additional email
notifications were sent at two-week intervals. Any non-responding institutions were then approached personally by members of the COG Outcomes and Survivorship, Adolescent and Young Adult, and
Nursing committees and invited to participate. personally by members of the COG Outcomes and Survivorship, Adolescent and Young Adult, and
Nursing committees and invited to participate. Results Representatives from 153 of the 209 invited COG institutions responded to the parent survey (73.2%
response rate). Of these, 137 institutions (89.5%) completed survey items about their healthcare
transition practices and were included in these analyses. Institution characteristics are outlined in Table
1. Data Analysis Frequencies were used to summarize institutional characteristics, healthcare transition practices, and
barriers to transition. Institutions were dichotomized based on whether or not they transferred patients
out for long-term follow-up care in adulthood. Institutions reporting their patients are "seen indefinitely
and not transferred elsewhere" on the institutional policies on age at transfer question were categorized
as not transferring patients. Institutions transferring patients at a certain age (i.e., age 18, 21, 25, 26 or
older) or transferring "when they are ready" without age-based limitations were categorized as
transferring patients. Associations between institutional characteristics (e.g., type and size) and transfer of young adult
patients were determined using chi-square tests. The total number of US Center for Health Care Transition
Improvement Core Elements offered at each COG institution was summed, and descriptive statistics were
used to characterize healthcare transition services. One-way ANOVAs and Tukey's HSD posthoc tests
were performed to compare differences between institutions on the number of healthcare transition
services provided. Kruskal-Wallis tests were also performed to confirm parametric results due to the
distribution of the number of healthcare transition services provided. Non-parametric posthoc tests were
adjusted by the Bonferroni correction for multiple tests. All analyses were conducted using SPSS
Statistics, Version 27.0 (IBM Corp, Armonk, NY), with p-values <0.05 considered statistically significant. Page 5/18 Page 5/18 Introduction of Healthcare Transition & Barriers to Transitioning Survivors Among the ninety-one institutions transferring survivors for adult long-term follow-up care (Table 2), the
concept of transition is often introduced to patients during pediatric survivorship care (89.0%) and less
frequently at earlier points during the cancer trajectory (e.g., diagnosis, during treatment, end of
treatment). The multidisciplinary staff typically involved in introducing survivors to the concept of
transition most frequently include survivorship care providers. The top three barriers to transitioning
survivors to adult care reported by institutions included: 1) perceived "lack of late-effects knowledge
among clinicians survivors are transferred to" (39.6%), 2) perceived "lack of survivor desire to leave
comfort of treating institution or oncologist" (31.9%), and 3) perceived "lack of survivor access to primary
care provider for reasons other than insurance such as geography" (26.4%). Transfer of Survivors & Models of Care Two-thirds of institutions (91/137) transfer survivors out of their pediatric programs to receive adult-
focused cancer survivorship care (Table 1). These transfers often occur following age-based institutional
policies at COG sites beginning at ≤18 years (8.0%), ≤21 years (13.1%), ≤25 years (7.3%), or ≥26 years
(12.4%). In comparison, 25.5% of institutions do not have age limitations related to transition and transfer
survivors "when they are ready." There was a significant relationship between institution type and
requirements to transfer young adult patients for survivor care, with freestanding children's hospitals
being more likely to transfer (81.4%) compared to other institution types (59.6%; p=.012, Cramer's
V=0.21). The percentage of institutions requiring transfer of care did not differ by program size (p=.084). Continuation of care at the pediatric center/treating institution (33.6%) and transfer to primary care
(33.6%) were equally common models of long-term follow-up care for young adult survivors of childhood
cancer (Table 1). Some sites (13.9%) reported transferring to adult oncology or adult cancer survivor
programs, while few (5.1%) reported typically transferring patients to specialty clinics for young adult-
aged survivors of childhood cancer. Associations with the Six Core Elements of Health Care Transition There were differences in the number of healthcare transition services offered based on institution
type (p=.002, η2=.16). Children's centers within adult hospitals offered fewer healthcare transition services
(M=0.82, SD=1.25) than freestanding children's hospitals (M=1.94, SD=1.55; p=.016) or children's
hospitals in close proximity to adult institutions (M=2.19, SD=1.63; p=.008). There was also a difference
in the number of transition services offered based on institution size (p=.019, η2=.13). The smallest There were differences in the number of healthcare transition services offered based on institution
type (p=.002, η2=.16). Children's centers within adult hospitals offered fewer healthcare transition services
(M=0.82, SD=1.25) than freestanding children's hospitals (M=1.94, SD=1.55; p=.016) or children's
hospitals in close proximity to adult institutions (M=2.19, SD=1.63; p=.008). There was also a difference
in the number of transition services offered based on institution size (p=.019, η2=.13). The smallest
programs (≤30 new oncology diagnoses per year) offered fewer transition services (M=0.54, SD=0.88)
than midsize programs with 61-90 new oncology diagnoses per year (M=2.11, SD=1.88; p=.034) or large
programs with >150 new oncology diagnoses per year (M=2.04, SD=1.57; p=.032). Non-parametric tests
yielded qualitatively equivalent findings for differences in the number of healthcare transition services
offered based on institution type and size. hospitals in close proximity to adult institutions (M=2.19, SD=1.63; p=.008). There was also a difference
in the number of transition services offered based on institution size (p=.019, η2=.13). The smallest
programs (≤30 new oncology diagnoses per year) offered fewer transition services (M=0.54, SD=0.88)
than midsize programs with 61-90 new oncology diagnoses per year (M=2.11, SD=1.88; p=.034) or large
programs with >150 new oncology diagnoses per year (M=2.04, SD=1.57; p=.032). Non-parametric tests
yielded qualitatively equivalent findings for differences in the number of healthcare transition services
offered based on institution type and size. Discussion The National Academy of Medicine and COG recommend continued survivor care throughout adulthood
because morbidity and mortality risks increase over time for long-term survivors of childhood cancer [2–
6]. Our results show that most COG institutions eventually transfer survivors of childhood cancer to
another institution for cancer-related follow-up care during adulthood. Sites vary in the age at which they
transfer care and the type of healthcare model used to deliver survivor care to adult-aged survivors. The National Academy of Medicine and COG recommend continued survivor care throughout adulthood
because morbidity and mortality risks increase over time for long-term survivors of childhood cancer [2–
6]. Our results show that most COG institutions eventually transfer survivors of childhood cancer to
another institution for cancer-related follow-up care during adulthood. Sites vary in the age at which they
transfer care and the type of healthcare model used to deliver survivor care to adult-aged survivors. Successful healthcare transition of adult-aged survivors of childhood cancer is critical for continued risk-
based screening and early detection of late effects in adulthood [3], yet very few programs deliver
comprehensive transition programming. No COG institutions in our sample reported offering healthcare
transition services in complete alignment with recommendations from the US Center for Health Care
Transition Improvement [16]. Taken together, these results suggest there is no current standard of care for
transitional survivorship care across COG institutions, and the development of best practices is needed to
guide transition program development for childhood cancer survivors. Within our sample of COG institutions that transferred adult-aged survivors, the most common model of
survivorship care for young adult survivors of childhood cancer was transfer to primary care providers
(PCPs). Previous studies have found that 73% of PCPs lack knowledge or training in survivorship care
[27], and only 25–37% report being comfortable caring for survivors of childhood cancer [28]. Respondents to the current and prior survey [26] of COG survivorship practices endorsed "lack of late-
effects knowledge among clinicians survivors are transferred to" as the most frequent barrier to
transitioning survivors to adult care. Correspondingly, young adult survivors of childhood cancer have
reported they perceive that their PCPs lack sufficient cancer knowledge and encounter difficulty finding
good/regular PCPs [29]. Alignment with the Six Core Elements of Health Care Transition Page 6/18
In regards to the US Center for Health Care Transition Improvement Six Core Elements, only a minority of
institutions reported having an established process to match and communicate with survivors' adult
providers (44.4%), incorporating transition readiness assessment into clinical practice (33.3%), having a
patient navigator or care coordinator to facilitate transition (28.6%), having a written transition policy
(24.2%), utilizing a transition readiness assessment tool (17.6%), and systematically obtaining feedback Page 6/18 from young adults about the transition process (8.8%; Table 2). On average, institutions reported offering
only one service aligned with any of the Six Core Elements (Median=1, M=1.56, SD=1.54), and no
institution offered all six services (range: 0-5; Figure 1). Discussion In efforts to bridge these gaps, recent recommendations from the American Within our sample of COG institutions that transferred adult-aged survivors, the most common model of
survivorship care for young adult survivors of childhood cancer was transfer to primary care providers
(PCPs). Previous studies have found that 73% of PCPs lack knowledge or training in survivorship care
[27], and only 25–37% report being comfortable caring for survivors of childhood cancer [28]. Respondents to the current and prior survey [26] of COG survivorship practices endorsed "lack of late-
effects knowledge among clinicians survivors are transferred to" as the most frequent barrier to
transitioning survivors to adult care. Correspondingly, young adult survivors of childhood cancer have
reported they perceive that their PCPs lack sufficient cancer knowledge and encounter difficulty finding
good/regular PCPs [29]. In efforts to bridge these gaps, recent recommendations from the American Page 7/18 Page 7/18 Academy of Pediatrics (AAP) encourage PCPs to incorporate the COG Long-Term Follow-Up Guidelines [4]
into their practice to deliver evidence-based surveillance to survivors [30]. These guidelines also
recommend PCPs collaborate with oncology subspecialists to educate survivors and ensure successful
transfer to adult-oriented survivor care. Future research is critically needed to evaluate the adoption and
implementation of these recommendations by primary care practices and the subsequent impacts on
survivors' healthcare transition outcomes. Our results indicate that the responsibilities of introducing healthcare transition and preparing patients
for transfer fall almost exclusively on survivorship programs, which are often under-resourced in terms of
funding and dedicated time for program development [25, 26]. These barriers likely contribute to the
concerning lack of transition programming offered at COG institutions observed in this study. Without
access to institutional programming to support transition preparation and transfer to adult care, it is not
surprising that most survivors of childhood cancer discontinue risk-based care during their young adult
years [9–11]. These findings underscore the need for improved organizational programming at the health
system level to better coordinate survivors' healthcare transition and improve access to and engagement
in lifelong risk-based survivor care. Pediatric oncology and survivor programs should consider implementing health system supports to
prepare survivors for healthcare transition and, if applicable, eventual transfer to adult-centered care. The
US Center for Health Care Transition Improvement offers guides to assist clinicians and health systems in
assessing their current healthcare transition activities and implementing the Six Core Elements of Health
Care Transition [https://www.gottransition.org/six-core-elements/]. Author Contributions All authors contributed to the study conception and design. Material preparation, data collection and
analysis were performed by Jordan Gilleland Marchak, Regine Haardörfer, Cam Escoffery, and Ann
Mertens. The first draft of the manuscript was written by Jordan Gilleland Marchak and all authors
commented on previous versions of the manuscript. All authors read and approved the final manuscript. Discussion These guides offer frameworks for
supporting the transition to adult-centered care with or without a change in survivor care provider; thus,
they are applicable regardless of whether or not institutions transfer survivors out for long-term follow-up
care in adulthood. Our results suggest COG programs are most in need of developing written transition
policies (Core Element 1- Transition and Care Policy/Guide), adopting transition readiness tools (Core
Element 3- Transition Readiness), and establishing mechanisms to obtain feedback from young adult
survivors about their transition process (Core Element 6- Transfer Completion). Although this study clearly characterizes healthcare transition practices and barriers at COG institutions,
it has important limitations to address in future research. Only 73% of COG institutions provided
responses to the entire survey. Due to our electronic recruitment methods, we cannot be certain if the
uncompleted surveys ever reached a potential respondent (e.g., out-of-date contact information listed in
COG directory, email blocked by firewall or delivered to spam, etc.) or if potential respondents were not
interested in sharing their program information. Most respondents completed the survey items sufficiently
to characterize their healthcare transition practices and were included in these analyses, but some
discontinued the survey early (n = 16) before transition questions were asked. Thus, these findings reflect
the state of healthcare transition programming at the majority of, but not all, COG institutions. Future
studies seeking to survey COG institutions may offer incentives to reimburse clinicians for their time
responding to the survey, embed very brief surveys into the recruitment email, or streamline survey items
in external data capture platforms using skip patterns to reduce respondent burden. For projects where Page 8/18 Page 8/18 surveying all COG institutions is not feasible, researchers may consider using purposeful sampling of
institutions based on program characteristics (i.e., size, location, type of institution) to enhance
generalizability. Competing Interests The authors have no relevant financial or non-financial interests to disclose. Conclusion Although most COG institutions transfer adult-aged survivors of childhood cancer elsewhere for survivor
care, very few programs deliver comprehensive programming to aid in successful healthcare transition. Successful healthcare transition can help promote increased early detection and treatment of late effects,
leading to reduced morbidity and mortality among adult survivors of childhood cancer. To better serve
survivors, novel research into best practices for survivor transition is needed, including transition
readiness tools and mechanisms to obtain survivor feedback about the transition process. Developing
successful strategies to support the implementation of healthcare transition programming in pediatric
oncology and collaborations with primary care practices will be integral to advancing lifelong health after
childhood cancer. Funding Research reported in this publication was supported by the Children's Oncology Group Outcomes and
Survivorship, Adolescent and Young Adult, and Nursing Committees, the National Cancer Institute under
Award Numbers R01CA218389, U10CA180886, and UG1CA189955, and the Intervention Development,
Dissemination, and Implementation Developing shared resource of Winship Cancer Institute of Emory
University and NIH/NCI under award number P30CA138292. The content is solely the responsibility of the
authors and does not necessarily represent the official views of the National Institutes of Health. Competing Interests Data Availability Following permissions from the Children's Oncology Group, the datasets generated during and/or
analyzed during the current study are available from the corresponding author on reasonable request. Page 9/18 Ethics approval This is an observational study. The Emory University's Institutional Review Board (IRB) determined that no
ethical approval is required. Acknowledgments: Research reported in this publication was supported by the Children's Oncology Group Outcomes and
Survivorship, Adolescent and Young Adult, and Nursing Committees, the National Cancer Institute under
Award Numbers R01CA218389, U10CA180886, and UG1CA189955, and the Intervention Development,
Dissemination, and Implementation Developing shared resource of Winship Cancer Institute of Emory
University and NIH/NCI under award number P30CA138292. The content is solely the responsibility of the
authors and does not necessarily represent the official views of the National Institutes of Health. References 1. SEER Cancer Statistics Review, 1975-2017 [Internet]. National Cancer Institute. Available from:
https://seer.cancer.gov/csr/1975_2017/. 2. Oeffinger KC, Mertens AC, Sklar CA, Kawashima T, Hudson MM, Meadows AT, Friedman DL, Marina N,
Hobbie W, Kadan-Lottick NS, Schwartz CL, Leisenring W, Robison LL. Chronic health conditions in
adult survivors of childhood cancer. N Engl J Med. 2006;355(15):1572-82. doi:
doi:10.1056/NEJMsa060185. PubMed PMID: 17035650; PMCID: Unavailable. 3. Hudson MM, Ness KK, Gurney JG, Mulrooney DA, Chemaitilly W, Krull KR, Green DM, Armstrong GT,
Nottage KA, Jones KE, Sklar CA, Srivastava DK, Robison LL. Clinical ascertainment of health
outcomes among adults treated for childhood cancer. JAMA. 2013;309(22):2371-81. doi:
10.1001/jama.2013.6296. PubMed PMID: 23757085; PMCID: PMC3771083. 3. Hudson MM, Ness KK, Gurney JG, Mulrooney DA, Chemaitilly W, Krull KR, Green DM, Armstrong GT,
Nottage KA, Jones KE, Sklar CA, Srivastava DK, Robison LL. Clinical ascertainment of health
outcomes among adults treated for childhood cancer. JAMA. 2013;309(22):2371-81. doi:
10.1001/jama.2013.6296. PubMed PMID: 23757085; PMCID: PMC3771083. 4. Children's Oncology Group. (October 2018). Long-term follow-up guidelines for survivors of
childhood, adolescent and young adult cancers. Version 5.0. Monrovia, CA. 4. Children's Oncology Group. (October 2018). Long-term follow-up guidelines for survivors of
childhood, adolescent and young adult cancers. Version 5.0. Monrovia, CA. 5. Institute of Medicine and the National Research Council. From cancer patient to cancer survivor: lost
in transition. Hewitt M, Greenfield, S., Stovall E., eds., editor. Washington, DC: National Academies
Press; 2006. 5. Institute of Medicine and the National Research Council. From cancer patient to cancer survivor: lost
in transition. Hewitt M, Greenfield, S., Stovall E., eds., editor. Washington, DC: National Academies
Press; 2006. 6. Mertens AC, Liu Q, Neglia JP, Wasilewski K, Leisenring W, Armstrong GT, Robison LL, Yasui Y. Cause-
specific late mortality among 5-year survivors of childhood cancer: the Childhood Cancer Survivor
Study. J Natl Cancer Inst. 2008;100(19):1368-79. doi: 10.1093/jnci/djn310. PubMed PMID:
18812549; PMCID: 2556702. Page 10/18
7. Schwartz L, Tuchman L, Hobbie W, Ginsberg J. A social-ecological model of readiness for transition
to adult-oriented care for adolescents and young adults with chronic health conditions. Child Care Page 10/18 Page 10/18 Health Dev. 2011;37(6):883-95. Health Dev. 2011;37(6):883-95. 8. Freyer DR. Transition of care for young adult survivors of childhood and adolescent cancer:
Rationale and approaches. J Clin Oncol. 2010;28(32):4810-8. Epub 2010 Mar 29. doi:
10.1200/JCO.2009.23.4278. PubMed PMID: 20351333; PMCID: PMC3018346. 9. References Oeffinger KC, Mertens AC, Hudson MM, Gurney JG, Casillas J, Chen H, Whitton J, Yeazel M, Yasui Y,
Robison LL. Health care of young adult survivors of childhood cancer: a report from the Childhood
Cancer Survivor Study. Ann Fam Med. 2004;2(1):61-70. Epub 2004/04/01. doi: 10.1370/afm.26. PubMed PMID: 15053285; PMCID: 1466633. 10. Ford JS, Tonorezos ES, Mertens AC, Hudson MM, Casillas J, Foster BM, Moskowitz CS, Smith SM,
Chou JF, Buchanan G, Robison LL, Oeffinger KC. Barriers and facilitators of risk-based health care for
adult survivors of childhood cancer: A report from the Childhood Cancer Survivor Study. Cancer. 2020;126(3):619-27. Epub 2019/10/19. doi: 10.1002/cncr.32568. PubMed PMID: 31626337; PMCID:
PMC6980215. 11. Nathan PC, Greenberg ML, Ness KK, Hudson MM, Mertens AC, Mahoney MC, Gurney JG, Donaldson
SS, Leisenring WM, Robison LL, Oeffinger KC. Medical care in long-term survivors of childhood
cancer: a report from the Childhood Cancer Survivor Study. J Clin Oncol. 2008;26(27):4401-9. doi:
10.1200/JCO.2008.16.9607. PubMed PMID: 18802152; PMCID: PMC2653112. 12. Henderson TO, Friedman DL, Meadows AT. Childhood cancer survivors: transition to adult-focused
risk-based care. Pediatrics. 2010;126(1):129-36. Epub 2010/06/16. doi: 10.1542/peds.2009-2802. PubMed PMID: 20547645; PMCID: Unavailable. 12. Henderson TO, Friedman DL, Meadows AT. Childhood cancer survivors: transition to adult-focused
risk-based care. Pediatrics. 2010;126(1):129-36. Epub 2010/06/16. doi: 10.1542/peds.2009-2802. PubMed PMID: 20547645; PMCID: Unavailable. 13. Reed-Knight B, Blount R, Gilleland J. The transition of health care responsibility from parents to youth
diagnosed with chronic illness: A developmental systems perspective. Fam Syst Health. 2014;32(2):219-34. doi: doi: 10.1037/fsh0000039 13. Reed-Knight B, Blount R, Gilleland J. The transition of health care responsibility from parents to youth
diagnosed with chronic illness: A developmental systems perspective. Fam Syst Health. 2014;32(2):219-34. doi: doi: 10.1037/fsh0000039 14. Schwartz LA, Brumley LD, Tuchman LK, et al. Stakeholder validation of a model of readiness for
transition to adult care. JAMA Pediatr. 2013;167(10):939-46. doi:
10.1001/jamapediatrics.2013.2223. PubMed PMID: 23959392; PMCID: PMC4289606. 14. Schwartz LA, Brumley LD, Tuchman LK, et al. Stakeholder validation of a model of readiness for
transition to adult care. JAMA Pediatr. 2013;167(10):939-46. doi:
10.1001/jamapediatrics.2013.2223. PubMed PMID: 23959392; PMCID: PMC4289606. 15. White PH, Cooley WC. Supporting the health care transition from adolescence to adulthood in the
medical home. Pediatrics. 2018;142(5). Epub 2018/10/24. doi: 10.1542/peds.2018-2587. PubMed
PMID: 30348754; PMCID: Unavailable. 15. White PH, Cooley WC. Supporting the health care transition from adolescence to adulthood in the
medical home. Pediatrics. 2018;142(5). Epub 2018/10/24. doi: 10.1542/peds.2018-2587. PubMed
PMID: 30348754; PMCID: Unavailable. 16. Cooley WC, Sagerman PJ. References Otth M, Denzler S, Koenig C, Koehler H, Scheinemann K. Transition from pediatric to adult follow-up
care in childhood cancer survivors-a systematic review. J Cancer Surviv. 2021;15(1):151-62. Epub
2020/07/18. doi: 10.1007/s11764-020-00920-9. PubMed PMID: 32676793; PMCID: Unavailable. 23. Otth M, Denzler S, Koenig C, Koehler H, Scheinemann K. Transition from pediatric to adult follow-up
care in childhood cancer survivors-a systematic review. J Cancer Surviv. 2021;15(1):151-62. Epub
2020/07/18. doi: 10.1007/s11764-020-00920-9. PubMed PMID: 32676793; PMCID: Unavailable. 24. Sadak KT, Szalda D, Lindgren BR, Kinahan KE, Eshelman-Kent D, Schwartz LA, Henderson T, Freyer
DR. Transitional care practices, services, and delivery in childhood cancer survivor programs: A
survey study of U.S. survivorship providers. Pediatr Blood Cancer. 2019;66(8):e27793. Epub
2019/05/18. doi: 10.1002/pbc.27793. PubMed PMID: 31099145; PMCID: PMC7521144. 25. Effinger KE, Haardörfer R, Marchak JG, Escoffery C, Landier W, Kommajosula A, Hendershot E, Sadak
KT, Eshelman-Kent D, Kinahan K, Freyer DR, Chow EJ, Mertens AC. Current pediatric cancer
survivorship practices: a report from the Children's Oncology Group. J Cancer Surviv. 2022:1-10. Epub
2022/02/01. doi: 10.1007/s11764-021-01157-w. PubMed PMID: 35098485; PMCID: PMC8801272. 25. Effinger KE, Haardörfer R, Marchak JG, Escoffery C, Landier W, Kommajosula A, Hendershot E, Sadak
KT, Eshelman-Kent D, Kinahan K, Freyer DR, Chow EJ, Mertens AC. Current pediatric cancer
survivorship practices: a report from the Children's Oncology Group. J Cancer Surviv. 2022:1-10. Epub
2022/02/01. doi: 10.1007/s11764-021-01157-w. PubMed PMID: 35098485; PMCID: PMC8801272. 26. Eshelman-Kent D, Kinahan K, Hobbie W, Landier W, Teal S, Friedman D, Nagarajan R, Freyer D. Cancer
survivorship practices, services, and delivery: a report from the Children's Oncology Group (COG)
nursing discipline, adolescent/young adult, and late effects committees. J Cancer Surviv. 2011;5(4):345-57. doi: 10.1007/s11764-011-0192-8. 26. Eshelman-Kent D, Kinahan K, Hobbie W, Landier W, Teal S, Friedman D, Nagarajan R, Freyer D. Cancer
survivorship practices, services, and delivery: a report from the Children's Oncology Group (COG)
nursing discipline, adolescent/young adult, and late effects committees. J Cancer Surviv. 2011;5(4):345-57. doi: 10.1007/s11764-011-0192-8. 27. Iyer NS, Mitchell HR, Zheng DJ, Ross WL, Kadan-Lottick NS. Experiences with the survivorship care
plan in primary care providers of childhood cancer survivors: a mixed methods approach. Support
Care Cancer. 2017;25(5):1547-55. Epub 2017/01/05. doi: 10.1007/s00520-016-3544-0. PubMed
PMID: 28050709; PMCID: Unavailable. 27. Iyer NS, Mitchell HR, Zheng DJ, Ross WL, Kadan-Lottick NS. Experiences with the survivorship care
plan in primary care providers of childhood cancer survivors: a mixed methods approach. Support
Care Cancer. 2017;25(5):1547-55. Epub 2017/01/05. doi: 10.1007/s00520-016-3544-0. PubMed
PMID: 28050709; PMCID: Unavailable. 28. References Supporting the health care transition from adolescence to adulthood in the
medical home. Pediatrics. 2011;128(1):182-200. Epub 2011/06/29. doi: 10.1542/peds.2011-0969. PubMed PMID: 21708806. 17. John AS, Jackson JL, Moons P, Uzark K, Mackie AS, Timmins S, Lopez KN, Kovacs AH, Gurvitz M. Advances in managing transition to adulthood for adolescents with congenital heart disease: A
practical approach to transition program design: A scientific statement from the American Heart
Association. J Am Heart Assoc. 2022;11(7):e025278. Epub 2022/03/18. doi:
10.1161/jaha.122.025278. PubMed PMID: 35297271; PMCID: PMC9075425. Page 11/18 Page 11/18 18. Society for Adolescent Health and Medicine. Transition to adulthood for youth with chronic
conditions and special health care needs. J Adolesc Health. 2020;66(5):631-4. Epub 2020/04/26. doi: 10.1016/j.jadohealth.2020.02.006. PubMed PMID: 32331627. 19. Bryant R, Porter JS, Sobota A, Association of Pediatric Hematology/Oncology N, American Society of
Pediatri Hematology O. APHON/ASPHO Policy Statement for the Transition of Patients With Sickle
Cell Disease From Pediatric to Adult Health Care. J Pediatr Oncol Nurs. 2015;32(6):355-9. Epub
2015/10/16. doi: 10.1177/1043454215591954. PubMed PMID: 26464507. 20. Rocque BG, Weprin BE, Blount JP, Hopson BD, Drake JM, Hamilton MG, Williams MA, White PH, Orrico
KO, Martin JE. Health care transition in pediatric neurosurgery: a consensus statement from the
American Society of Pediatric Neurosurgeons. J Neurosurg: Pediatr. 2020:1-9. Epub 2020/02/15. doi:
10.3171/2019.12.Peds19524. PubMed PMID: 32059192. 20. Rocque BG, Weprin BE, Blount JP, Hopson BD, Drake JM, Hamilton MG, Williams MA, White PH, Orrico
KO, Martin JE. Health care transition in pediatric neurosurgery: a consensus statement from the
American Society of Pediatric Neurosurgeons. J Neurosurg: Pediatr. 2020:1-9. Epub 2020/02/15. doi:
10.3171/2019.12.Peds19524. PubMed PMID: 32059192. 21. Ginsberg JP, Hobbie WL, Carlson CA, Meadows AT. Delivering long-term follow-up care to pediatric
cancer survivors: transitional care issues. Pediatr Blood Cancer. 2006;46(2):169-73. Epub
2005/12/02. doi: 10.1002/pbc.20610. PubMed PMID: 16320315; PMCID: Unavailable. 21. Ginsberg JP, Hobbie WL, Carlson CA, Meadows AT. Delivering long-term follow-up care to pediatric
cancer survivors: transitional care issues. Pediatr Blood Cancer. 2006;46(2):169-73. Epub
2005/12/02. doi: 10.1002/pbc.20610. PubMed PMID: 16320315; PMCID: Unavailable. 22. Nathan PC, Hayes-Lattin B, Sisler JJ, Hudson MM. Critical issues in transition and survivorship for
adolescents and young adults with cancers. Cancer. 2011;117(10 Suppl):2335-41. Epub 2011/05/20. doi: 10.1002/cncr.26042. PubMed PMID: 21523755; PMCID: Unavailable. 22. Nathan PC, Hayes-Lattin B, Sisler JJ, Hudson MM. Critical issues in transition and survivorship for
adolescents and young adults with cancers. Cancer. 2011;117(10 Suppl):2335-41. Epub 2011/05/20. doi: 10.1002/cncr.26042. PubMed PMID: 21523755; PMCID: Unavailable. 23. References Suh E, Daugherty CK, Wroblewski K, Lee H, Kigin ML, Rasinski KA, Ford JS, Tonorezos ES, Nathan PC,
Oeffinger KC, Henderson TO. General internists' preferences and knowledge about the care of adult 28. Suh E, Daugherty CK, Wroblewski K, Lee H, Kigin ML, Rasinski KA, Ford JS, Tonorezos ES, Nathan PC,
Oeffinger KC, Henderson TO. General internists' preferences and knowledge about the care of adult 28. Suh E, Daugherty CK, Wroblewski K, Lee H, Kigin ML, Rasinski KA, Ford JS, Tonorezos ES, Nathan PC,
Oeffinger KC, Henderson TO. General internists' preferences and knowledge about the care of adult Page 12/18 Page 12/18 survivors of childhood cancer: a cross-sectional survey. Ann Intern Med. 2014;160(1):11-7. Epub
2014/02/28. doi: 10.7326/m13-1941. PubMed PMID: 24573662; PMCID: PMC4337806. 29. Signorelli C, Wakefield CE, Fardell JE, Foreman T, Johnston KA, Emery J, Thornton‐Benko E, Girgis A,
Lie HC, Cohn RJ. The role of primary care physicians in childhood cancer survivorship care:
Multiperspective interviews. Oncologist. 2018. Epub Aug 31 2018. doi: 10.1634/theoncologist.2018-
0103 PubMed PMID: 30171066; PMCID: PMC6516111. 30. Hudson MM, Bhatia S, Casillas J, Landier W. Long-term follow-up care for childhood, adolescent, and
young adult cancer survivors. Pediatrics. 2021;148(3). Epub 2021/09/01. doi: 10.1542/peds.2021-
053127. PubMed PMID: 34462344; PMCID: Unavailable. Tables 30. Hudson MM, Bhatia S, Casillas J, Landier W. Long-term follow-up care for childhood, adolescent, and
young adult cancer survivors. Pediatrics. 2021;148(3). Epub 2021/09/01. doi: 10.1542/peds.2021-
053127. PubMed PMID: 34462344; PMCID: Unavailable. 29. Signorelli C, Wakefield CE, Fardell JE, Foreman T, Johnston KA, Emery J, Thornton‐Benko E, Girgis A,
Lie HC, Cohn RJ. The role of primary care physicians in childhood cancer survivorship care:
Multiperspective interviews. Oncologist. 2018. Epub Aug 31 2018. doi: 10.1634/theoncologist.2018-
0103 PubMed PMID: 30171066; PMCID: PMC6516111. survivors of childhood cancer: a cross-sectional survey. Ann Intern Med. 2014;160(1):11-7. Epub
2014/02/28. doi: 10.7326/m13-1941. PubMed PMID: 24573662; PMCID: PMC4337806. Tables Page 13/18 Table 1
Characteristics of Children's Oncology Group (COG) institutions reporting on healthcare transition services
for survivors (n = 137)
Institution characteristics
N (%)
Type of institution
Children's Hospital in Close Proximity to Adult Institution
33
(24.4%)
Children's Center Within Adult Hospital
53
(39.3%)
Freestanding Children's Hospital
43
(31.9%)
Community Hospital/Clinic
6 (4.5%)
Missing
2 (1.5%)
Institution size
≤ 30 new oncology diagnoses per year
19
(13.9%)
31–60 new oncology diagnoses per year
34
(24.8%)
61–90 new oncology diagnoses per year
30
(21.9%)
91–150 new oncology diagnoses per year
19
(13.9%)
> 150 new oncology diagnoses per year
27
(19.7%)
Upper age limit for newly diagnosed patients to begin treatment
18 years
19
(13.9%)
> 18–21 years
50
(36.5%)
> 21–29 years
49
(35.8%)
30 + years
12
(8.8%)
Other
6 (4.4%)
Missing
1 (0.7%) Institution characteristics
N (%)
Patients are seen indefinitely and not transferred elsewhere
46
(33.6%)
No age limitation, survivors transitioned when they are ready
35
(25.5%)
Transfer at age 18
11
(8.0%)
Transfer at age 21
18
(13.1%)
Transfer at age 25
10
(7.3%)
Transfer at age 26 or older
17
(12.4%)
Models of survivorship care for young adult survivors of childhood cancer
Patients continue to be seen indefinitely at pediatric center/treating institution and are not
transferred elsewhere
46
(33.6%)
Transfer to adult oncology or adult survivorship program
19
(13.9%)
Transfer to primary care provider
46
(33.6%)
Transfer to specialty clinic for adult survivors of childhood cancer
7 (5.1%)
Different adult care options provided based on health risks and/or survivor preference
7 (5.1%)
No specific model of transfer to adult-based care
9 (6.6%)
Missing
3 (2.2%) Institution characteristics
N (%)
Patients are seen indefinitely and not transferred elsewhere
46
(33.6%)
No age limitation, survivors transitioned when they are ready
35
(25.5%)
Transfer at age 18
11
(8.0%)
Transfer at age 21
18
(13.1%)
Transfer at age 25
10
(7.3%)
Transfer at age 26 or older
17
(12.4%)
Models of survivorship care for young adult survivors of childhood cancer
Patients continue to be seen indefinitely at pediatric center/treating institution and are not
transferred elsewhere
46
(33.6%)
Transfer to adult oncology or adult survivorship program
19
(13.9%)
Transfer to primary care provider
46
(33.6%)
Transfer to specialty clinic for adult survivors of childhood cancer
7 (5.1%)
Different adult care options provided based on health risks and/or survivor preference
7 (5.1%)
No specific model of transfer to adult-based care
9 (6.6%)
Missing
3 (2.2%) Institution characteristics Page 15/18 Table 2
Healthcare transition practices and barriers among Children's Oncology Group (COG) institutions with
requirements to transfer survivors for adult long-term follow-up care (n = 91)
Healthcare Transition Programming
N (%)
Timing of introducing survivors to the concept of transition to adult care*
Transition is consistently introduced at diagnosis
1
(1.1%)
Transition is consistently introduced during treatment
5
(5.5%)
Transition is consistently introduced at end of treatment
19
(20.9%)
Transition is consistently introduced in survivorship program
81
(89.0%)
Staff involved in introducing survivors to the concept of transition to adult care*
Survivorship Physician
64
(70.3%)
Survivorship Advanced Practice Provider
51
(56.0%)
Survivorship Social Worker
23
(25.3%)
Survivorship Nurse Coordinator
20
(22.0%)
Survivorship Psychologist
5
(5.5%)
Non-survivorship team member
11
(12.1%)
US Center for Health Care Transition Improvement Core Elements offered at institution*
Program has written transition policy/statement
22
(24.2%)
Program has adopted a transition readiness tool for use
16
(17.6%)
Program has incorporated transition readiness assessment into clinical practice
30
(33.0%)
Program has a navigator/care coordinator to facilitate transition
26
(28.6%)
Program has established process to match and communicate with survivors' selected
40 Table 2
Healthcare transition practices and barriers among Children's Oncology Group (COG) institutions with
requirements to transfer survivors for adult long-term follow-up care (n = 91)
Healthcare Transition Programming
N (%)
Timing of introducing survivors to the concept of transition to adult care*
Transition is consistently introduced at diagnosis
1
(1.1%)
Transition is consistently introduced during treatment
5
(5.5%)
Transition is consistently introduced at end of treatment
19
(20.9%)
Transition is consistently introduced in survivorship program
81
(89.0%)
Staff involved in introducing survivors to the concept of transition to adult care*
Survivorship Physician
64
(70.3%)
Survivorship Advanced Practice Provider
51
(56.0%)
Survivorship Social Worker
23
(25.3%)
Survivorship Nurse Coordinator
20
(22.0%)
Survivorship Psychologist
5
(5.5%)
Non-survivorship team member
11
(12.1%)
US Center for Health Care Transition Improvement Core Elements offered at institution*
Program has written transition policy/statement
22
(24.2%)
Program has adopted a transition readiness tool for use
16
(17.6%)
Program has incorporated transition readiness assessment into clinical practice
30
(33.0%)
Program has a navigator/care coordinator to facilitate transition
26
(28.6%)
Program has established process to match and communicate with survivors' selected
40 Healthcare Transition Programming
N (%)
Program systematically obtains feedback from young adult survivors about the transition
process
8
(8.8%)
Barriers to Transitioning Survivors to Adult Care†
N (%)
Lack of late-effects knowledge among clinicians survivors are transferred to
36
(39.6%)
Lack of survivor desire to leave comfort of treating institution/oncologist
29
(31.9%)
Lack of survivor access to primary care provider for reasons other than insurance such as
geography
24
(26.4%)
Survivors' lack of healthcare insurance or insurance limitations
23
(25.3%)
Survivor knowledge deficit about importance of maintaining long-term follow-up care in
adulthood
19
(20.9%)
Conflict between COG recommendations for risk-based follow-up and adult health care
provider recommendations
10
(11.0%)
Lack of oncology provider desire to "let go" of survivors
10
(11.0%)
Program does not have any barriers
9
(9.9%)
*Respondents instructed to check all that apply; †Respondents instructed to choose top 2 barriers Healthcare Transition Programming Figures Page 17/18 Page 17/18 Figure 1 Legend not included with this version Page 18/18 Page 18/18
|
https://openalex.org/W3119745711
|
https://www.researchsquare.com/article/rs-71178/latest.pdf
|
English
| null |
QTL Detection for Bread Wheat Processing Quality in A Nested Association Mapping Population of Semi-Wild and Domesticated Wheat Varieties
|
Research Square (Research Square)
| 2,020
|
cc-by
| 8,847
|
QTL detection for bread wheat processing quality in a nested
association mapping population of semi-wild and domesticated
wheat varieties QTL detection for bread wheat processing quality in a nested
association mapping population of semi-wild and domesticated
wheat varieties QTL detection for bread wheat processing quality in a nested
association mapping population of semi-wild and domesticated
wheat varieties
Junmei Hu
Shandong Agricultural University
Guilian Xiao
Shandong Agricultural University
Peng Jiang
Shandong Agricultural University
Yan Zhao
Shandong Agricultural University
Guangxu Zhang
Lianyungang Academy of Agricultural Sciences
Xin Ma
Shandong Agricultural University
Jie Yao
Yantai Academy of Agricultural Sciences in Shandong Province
Lixia Xue
Agricultural Technology Station, Sunwu Sub-district Office, Huimin Country, Shandong Province
Peisen Su
Liaocheng University
Yinguang Bao
(
baoyinguang@163.com
)
Shandong Agricultural University
Research article
Keywords: Wheat, Processing quality, Quantitative trait locus, Semi-wild wheat, Nested association mapping (NAM) population
Posted Date: December 8th, 2020
DOI: https://doi.org/10.21203/rs.3.rs-71178/v3
License:
This work is licensed under a Creative Commons Attribution 4.0 International
License.
Read Full License QTL detection for bread wheat processing quality in a nested
association mapping population of semi-wild and domesticated
wheat varieties
Junmei Hu
Shandong Agricultural University
Guilian Xiao
Shandong Agricultural University
Peng Jiang
Shandong Agricultural University
Yan Zhao
Shandong Agricultural University
Guangxu Zhang
Lianyungang Academy of Agricultural Sciences
Xin Ma
Shandong Agricultural University
Jie Yao
Yantai Academy of Agricultural Sciences in Shandong Province
Lixia Xue
Agricultural Technology Station, Sunwu Sub-district Office, Huimin Country, Shandong Province
Peisen Su
Liaocheng University
Yinguang Bao
(
baoyinguang@163.com
)
Shandong Agricultural University
Research article
Keywords: Wheat, Processing quality, Quantitative trait locus, Semi-wild wheat, Nested association mapping (NAM) population
Posted Date: December 8th, 2020
DOI: https://doi.org/10.21203/rs.3.rs-71178/v3
License:
This work is licensed under a Creative Commons Attribution 4.0 International
License. Read Full License Background Bread wheat (Triticum aestivum L.) is a crucial source of protein, minerals and vitamins that feeds over 35% of the world’s population
[1, 2]. Hence, improving the processing quality of wheat is an important goal in wheat breeding. Investigation of the relationship
between seed storage protein alleles and processing characteristics indicates that storage proteins are the main determinant of wheat
processing quality [3]. The seed storage proteins in wheat consist of gliadin and glutenin, which can be used to predict dough
rheological properties, including the viscoelastic and mixing properties [4]. The genes coding for gliadin are located on the short arms
of the chromosomes of homologous groups 1 and 6 [5]. Glutenin in the endosperm consists of high-molecular-weight (HMW-GS) and
low-molecular-weight glutenin subunits (LMW-GS). HMW-GSs are encoded by Glu-1 loci that are located on the long arms of the
chromosomes of homologous group 1, including Glu-A1, Glu-B1 and Glu-D1 [5]. LMW-GSs are encoded by Glu-3 loci that are located on
the short arms of the chromosomes of group 1, including Glu-A3, Glu-B3 and Glu-D3 [6]. Research has indicated that both glutenin and
gliadin are significantly associated with wheat processing quality by affecting the viscoelasticity and flexibility of dough [7, 8]. Most of the quality-related traits of interest in wheat breeding are characterized by polygenic inheritance, which is generally studied with
quantitative trait loci (QTL) mapping [3, 9–16]. Some stable QTL for protein content were discovered on chromosomes 1A, 1D, 2B, 3A,
4A, 5A, 5D, 7A, and 7B [3, 7, 12, 15, 17, 18]. Krystkowiak et al. [7] detected one major QTL on chromosome 5D that influences starch
content, wet gluten content, and zeleny sedimentation value. Stable QTL for the starch content of wheat flour were detected on
chromosomes 4A and 7D [9]. In addition, QTL for wet gluten content of wheat flour were identified on chromosomes 5AS and 5AL [15]. Due to a complex interaction between proteins and other components, such as pentosans, the predictability of dough strength from
chemical composition is difficult, and therefore rheological tests are required [19, 20]. The dough rheological characteristics of wheat
are quantitative traits that are dependent on multiple genes and are greatly influenced by environmental conditions. Studies have
reported that dough properties are influenced by the properties of storage proteins, which can be reflected by mixograph, farinograph,
and extensograph parameters [13, 19, 21]. Research article License:
This work is licensed under a Creative Commons Attribution 4.0 International
License. Read Full License Page 1/16 Page 1/16 Abstract Background: Wheat processing quality is an important factor in evaluating overall wheat quality, and dough characteristics are
important when assessing the processing quality of wheat. As a notable germplasm resource, semi-wild wheat has a key role in the
study of wheat processing quality. Results: In this study, four dough rheological characteristics were collected in four environments using a nested association mapping
(NAM) population consisting of semi-wild and domesticated wheat varieties to identify quantitative trait loci (QTL) for wheat
processing quality. A total of 49 QTL for wheat processing quality were detected, explaining 0.36–10.82% of the phenotypic variation. These QTL were located on all wheat chromosomes except for 2D, 3A, 3D, 6B, 6D and 7D. Compared to previous studies, 29 QTL were
newly identified. Four novel QTL, QMlPH-1B.4, QMlPH-3B.4, QWdEm-1B.2 and QWdEm-3B.2, were stably identified in three or more
environments, among which QMlPH-3B.4 was a major QTL. Moreover, eight important genetic regions for wheat processing quality
were identified on chromosomes 1B, 3B and 4D, which showed pleiotropy for dough characteristics. In addition, out of 49 QTL, 15
favorable alleles came from three semi-wild parents, suggesting that the QTL alleles provided by the semi-wild parent were not utilized
in domesticated varieties. Conclusions: The results show that semi-wild wheat varieties can enrich the existing wheat gene pool and provide broader variation
resources for wheat genetic research. Background Many rheological tests have been widely used as predictors of wheat processing quality and
end-use quality [3, 15, 19]. Mann et al. [3] discovered that dough rheology QTL were highly correlated across multiple environments and
primarily influenced by the Glu-1 loci (Glu-B1, Glu-D1). Tsilo et al. [22] detected a major QTL cluster for dough rheological properties on
chromosome 1B, which explained the large total phenotypic variation in dough development time, mixing tolerance index, dough
stability and time to dough breakdown. Page 2/16
Several segregant biparental populations can be adopted for QTL mapping, such as backcross, F2, doubled haploid (DH), introgression
lines and recombinant inbred line populations. In most previous studies, these approaches for wheat processing quality QTL mapping
were utilized [9, 10, 23]. However, the linkage analysis achievable with biparental populations showed a narrow genetic background and
was often able to detect QTL only with large intervals because of limited recombination events [24-26]. This limitation can be partially
overcome by analyzing multiple related populations, such as nested association mapping (NAM) population [27, 28]. Nested
association mapping populations created by the hybridization of one common parent with several other parents have become desired Page 2/16 Page 2/16 for QTL analyses due to their advantages of broad genetic diversity and high resolution. Such populations have been used to identify
QTL in different crops, including maize, soybean, sorghum, barley, bread wheat and durum wheat [29, 30]. However, there are few
studies on QTL analysis of wheat processing quality using a NAM population [31, 32]. In previous studies, most of the material on wheat processing quality represented domesticated cultivars, and compared with wild and
semi-wild varieties, their genetic diversity will decrease with domestication [33]. Through whole-genome sequencing analysis, it was
determined that composite introgression from wild populations contributed to 4–32% of the bread wheat genome, which increased the
genetic diversity of bread wheat [34]. Three semi-wild wheat subspecies germplasm resources unique to western China, including the
Tibetan weedrace (T. aestivum ssp. tibetanum Shao) characterized by strong seed dormancy, hulled glumes and brittle spikelets,
Xinjiang rice wheat (T. petropavlovskyi Udats. et Migusch.) characterized by a long glume, and Yunnan hulled wheat, or “Tiekemai” (T. aestivum ssp. yunnanense King), named for its very hard and tough glumes that adhere to the grains [35-37]. Background The unique semi-wild
wheat subspecies in China have a primitive chromosomal constitution, which is crucial to probe the effect of domestication on
processing quality in wheat breeding [35]. Therefore, in this study, a wheat NAM population was constructed by crossing one common
parent, Yanzhan 1, and four divergent parents, including three semi-wild cultivars from China and one domesticated variety from the
British islands. This NAM population was used for QTL mapping for wheat processing quality, which will facilitate high-quality wheat
breeding and marker-assisted selection (MAS) in wheat breeding. Phenotypic analysis of wheat processing quality The five parents of the NAM population had different dough rheological characteristics (Fig. 1A). Among them, HU (Hussar) had the
best dough characteristics, while the dough characteristics of YZ (Yanzhan 1) and YN (Yunnanxiaomai) were poor. We found that the
common parent YZ had a longer MlPT (midline peak time), wider PkWd (peak width) and wider WdEm (width at eight minutes) than the
other parents, CY (Chayazheda 29) and YN, and had the smallest MlPH (midline peak height) compared with the other four donor
parents. HU had the highest values for all dough rheological characteristics compared with the other four parents, which indicated that
HU had optimal dough characteristics. CY had a longer MlPT, wider PkWd and wider WdEm than YN. YN had a wider MlPH than YZ, CY
and YT but the shortest MlPT and the narrowest WdEm compared with the other four parents. Compared with YZ, CY and YN, YT
(Yutiandaomai) had the longest MlPT, widest WdEm and narrowest PkWd. Observing the phenotypic data distribution of the NAM
population, there was strong transgressive segregation for all dough rheological characteristics in each RIL population except for the
MlPT of HU-RILs (Fig. 1B, Table S1). To evaluate the pairwise correlations between dough rheological characteristics, Pearson’s correlation was estimated using BLUP (best
linear unbiased prediction) values combined over four environments (Fig. 1C). WdEm was significantly positively correlated with MlPT
and PkWd but significantly negatively correlated with MlPH. PkWd was significantly positively correlated with MlPT and MlPH. The
correlation between MlPT and MlPH was not significant. The heritabilities of dough rheological characteristics in the NAM population were 42.7–84.7%, and they were differed largely in the
four RIL populations (Table S2). PKWD is a relatively important dough rheological parameter to measure wheat processing quality, and
its phenotype is greatly affected by the environment (42.7 –59.7%). Among them, the heritabilities of the HU-RIL and YT-RIL populations
were higher, whereas those in the YN-RIL population were lower. QTL analysis of wheat processing quality QMlPT-1B.1, QMlPT-3B.2 and QMlPT-5A, with LOD values of 3.71–6.38, 3.08–
8.03 and 2.66–4.88, respectively, were found in two environments, accounting for 2.24–5.42%, 2.07–3.70% and 1.78–1.82% of the
phenotypic variation, respectively. The favorable alleles of two and three QTL of the ten QTL were provided by semi-wild parents YN
and YT, respectively, while one and four QTL out of the remaining QTL were provided by domesticated parents HU and YZ, respectively
(Table S4). For MlPH, 18 QTL were identified, which were distributed on chromosomes 1B (6), 2B, 3B (5), 4D, 5B (2), 5D, 6A, and 7B in four
environments and the combined environment analysis, explaining 0.36–10.82% of the phenotypic variation (Fig. 2, Table S3). Twelve
of the eighteen QTL were detected in both individual environments and the combined environment analysis. The favorable alleles of
two stable QTL (QMlPH-3B.3 and QMlPH-3B.4) were contributed by HU, which were identified in all four environments and the
combined environment analysis, explaining 1.50–8.60% and 1.71–10.82% of the phenotypic variation, respectively. QMlPH-1B.4, a
favorable allele from the common parent YZ, was stably identified in four environments, with an LOD value of 4.01–9.75 and
phenotypic variation of 1.42–4.50%. QMlPH-1B.3 and QMlPH-5D, with LOD values of 5.18–10.25 and 3.21–5.99, respectively, were
stably detected in the two environments and the combined environment analysis, accounting for 1.15–4.90% and 1.10–2.21% of the
phenotypic variation, respectively. Eight QTL, including QMlPH-1B.5, QMlPH-1B.6, QMlPH-3B.1, QMlPH-4D, QMlPH-5B.1, QMlPH-5B.2,
QMlPH-6A and QMlPH-7B, were detected in one environment and the combined environment analysis. In addition, five QTL, QMlPH-
1B.1, QMlPH-1B.2, QMlPH-2B, QMlPH-3B.2 and QMlPH-3B.5, were identified in two environments. Three OTL with favorable alleles were
detected in the three semi-wild parents CY, YN, and YT (Table S4). Compared with the other four parents, the alleles of the domesticated
parent HU increased the MlPH for five QTL, while the common parent YZ decreased the MlPH for ten QTL. For PkWd, eleven QTL were stably detected on chromosomes 1A, 1B (4), 2B, 3B (2), 4A, 6A, and 7A, explaining 0.55–6.09% of the
phenotypic variation (Fig. 2, Table S3). All of these QTL except for QPkWd-1B.1 were identified in both individual environments and the
combined environment analysis. QPkWd-3B.1 and QPkWd-3B.2 are favorable alleles from the common parent YZ and were detected in
three or more environments. QTL analysis of wheat processing quality For five QTL (QPkWd-1B.2, QPkWd-1B.3, QPkWd-1B.4, QPkWd-4A and QPkWd-6A), the alleles increasing
PkWd were provided by YZ, accounting for 1.74–5.31%, 1.85–4.70%, 1.93–4.85%, 0.76–3.02% and 1.49–3.37% of the phenotypic
variation, respectively. For QPkWd-1A and QPkWd-7A, the alleles increasing PkWd were donated by HU, with 5.85–7.29 and 3.04–3.36
of the LOD value, respectively, accounting for 2.15–2.81% and 1.12–2.88% of the phenotypic variation, respectively. In addition, in
QPkWd-1B.1 and QPkWd-2B, the alleles decreasing PkWd were provided by CY and YT, respectively, accounting for 0.96–4.80% and
0.55–4.16% of the phenotypic variation, respectively. For WdEm, ten QTL were detected on chromosomes 1B (3), 1D, 2A, 2B, 3B (2), 4D, and 5B in individual environmental and combined
environment analyses, accounting for 0.51–9.70% of the phenotypic variation (Fig. 2, Table S3). Six QTL were identified in both
individual environments and the combined environment analysis. Two QTL, QWdEm-1B.2 and QWdEm-3B.1, were stably identified in
three environments and the combined environment analysis, with 4.70–5.78 and 4.44–16.13 of the LOD value, respectively, explaining
0.51–2.42% and 2.14–8.82% of the phenotypic variation, respectively. QWdEm-1B.1 and QWdEm-1B.3 had LOD values of 4.14–4.89
and 4.87–6.87, respectively and were detected in the two environments and the combined environment analysis. In these two QTL, the
alleles decreasing WdEm were donated by YN, accounting for 1.19–2.73% and 1.75–3.25% of the phenotypic variation, respectively. The alleles in HU increased WdEm for two QTL, QWdEm-2A and QWdEm-4D, which explained 2.34–4.05% and 0.85–2.12% of the
phenotypic variation, respectively. For QWdEm-1D and QWdEm-2B, favorable alleles were donated by YT and YN, respectively. Favorable alleles of three QTL, QWdEm-3B.1, QWdEm-3B.2, and QWdEm-5B, were provided by the common parent YZ. Among the ten
QTL for WdEm, the favorable alleles of four and one QTL were provided by semi-wild parents YN and YT, respectively. In addition, two
and three QTL were provided by domesticated parents HU and YZ, respectively (Table S4). Favorable alleles of three QTL, QWdEm-3B.1, QWdEm-3B.2, and QWdEm-5B, were provided by the common parent YZ. Among the ten
QTL for WdEm, the favorable alleles of four and one QTL were provided by semi-wild parents YN and YT, respectively. In addition, two
and three QTL were provided by domesticated parents HU and YZ, respectively (Table S4). QTL analysis of wheat processing quality A total of 49 QTL were detected on chromosomes 1A (2), 1B (17), 1D, 2A, 2B (3), 3B (11), 4A, 4B, 4D (2), 5A, 5B (3), 5D, 6A (2), 7A, and
7B (2) for wheat processing quality in four individual environments and combined QTL analysis (Fig. 2, Table S3). Ten, eighteen, eleven,
and ten QTL were identified for MlPT, MlPH, PkWd and WdEm, respectively. These QTL explained 0.36–10.82% of the phenotypic
variation. Thirty-four of these QTL were identified in the individual environment and the combined environment analysis. The favorable
alleles of two, seven, six, ten and twenty-four QTL were provided by parents CY, YN, YT, HU and YZ, respectively (Table S4). For MlPT, ten QTL were found on chromosomes 1A, 1B (4), 3B (2), 4B, 5A, and 7B in four environments and the combined environment
analysis, explaining a range of 1.47% to 8.29% of the phenotypic variation (Fig. 2, Table S3). Six of those QTL were detected in the
individual environment and combined environment analyses. One stable QTL, QMlPT-1B.2, had a favorable allele from YT and was
found in two individual environments and the combined environment analysis, explaining 2.20–3.66% of the phenotypic variation. The Page 3/16 Page 3/16 donor parent YN contributed the best favorable allele for QMlPT-3B.1, which was stably detected in three individual environments,
explaining 2.86–4.14% of the phenotypic variation. Five QTL, including QMlPT-1A, QMlPT-1B.3, QMlPT-1B.4, QMlPT-4B and QMlPT-7B,
were identified in one environment and the combined environment analysis, with 4.15–8.29%, 2.80–3.28%, 3.31–4.19%, 2.74–3.11%
and 1.47–4.02% of the phenotypic variation, respectively. QMlPT-1B.1, QMlPT-3B.2 and QMlPT-5A, with LOD values of 3.71–6.38, 3.08–
8.03 and 2.66–4.88, respectively, were found in two environments, accounting for 2.24–5.42%, 2.07–3.70% and 1.78–1.82% of the
phenotypic variation, respectively. The favorable alleles of two and three QTL of the ten QTL were provided by semi-wild parents YN
and YT, respectively, while one and four QTL out of the remaining QTL were provided by domesticated parents HU and YZ, respectively
(Table S4). donor parent YN contributed the best favorable allele for QMlPT-3B.1, which was stably detected in three individual environments,
explaining 2.86–4.14% of the phenotypic variation. Five QTL, including QMlPT-1A, QMlPT-1B.3, QMlPT-1B.4, QMlPT-4B and QMlPT-7B,
were identified in one environment and the combined environment analysis, with 4.15–8.29%, 2.80–3.28%, 3.31–4.19%, 2.74–3.11%
and 1.47–4.02% of the phenotypic variation, respectively. Comparison with previous studies In this study, 49 QTL for wheat processing quality were identified, 29 of which were unique to this study compared with previous
studies, and four novel QTL (QMlPH-1B.4, QMlPH-3B.4, QWdEm-1B.2 and QWdEm-3B.2) were stably identified in three or more
environments (Fig. 2, Tables 2, S3). For MlPT, five of ten QTL were previously reported. QMlPT-1A, with a favorable allele in HU, was
mapped close to the gene Glu-A1, whose effect was consistent with the longer MlPT of Hussar (Figs. 1A, 2, Table S3) [38]. Similar
genetic regions of two QTL, QMlPT-1B.3 and QMlPT-1B.4, which had favorable alleles from the common parent YZ, were reported by
Tsilo et al. [22] (Fig. 2, Table S3). QMlPT-3B.1 was previously reported by Liu et al. [11] to be located at a similar genetic region on
chromosome 3B. QMlPT-7B was identified close to the Psy-B1 gene, which indicates that Psy-B1 is associated with the synthesis of
carotenoids that are not related to dough characteristics [39]. Eighteen QTL for MlPH were identified, five of which were previously reported (Fig. 2, Table S3). QMlPH-1B.1 was detected in two
environments and was located near two genes, Gli-B1 and Glu-B3. QMlPH-1B.5 was located near gene Glu-B1 [40]. Three QTL, QMlPH-
3B.2, QMlPH-3B.3, and QMlPH-6A, were detected at a similar genetic region by Carter et al. [41], Liu et al. [11], and Li et al. [23],
respectively. Among the five QTL that were reported in previous studies, the allele of QMlPH-3B.3 was provided by HU with higher MlPH,
and favorable alleles of the other QTL were detected in YZ with lower MlPH, indicating that YZ may have a recessive allelic variation
gene that affects gluten strength (Fig. 1A, Table S3). Regarding PkWd, eleven QTL were detected, six of which were reported by previous studies (Fig. 2, Table S3). Two QTL, QPkWd-1B.1
and QPkWd-4A, at similar genetic regions were reported by Li et al. [23, 42]. QPkWd-1B.3 was mapped close to the gene Glu-B1 [40]. Two QTL, QPkWd-2B and QPkWd-3B.1, were previously reported by Liu et al. [11] in a similar genetic region. In addition, QPkWd-7A was
located at a similar genetic region in a previous study by Zhang et al. [15]. Regarding WdEm, four of ten QTL were detected in previous studies (Fig. 2, Table S3). Three QTL, QWdEm-1B.1, QWdEm-1D, and
QWdEm-3B.1, were reported from similar genetic regions on chromosomes 1B, 1D, and 3B, respectively [11, 23, 42]. Trait correlations Wheat processing quality is a quantitative genetic trait controlled by multiple genes and affected by the environment. The dough
characteristics influenced by glutelin and gliadin are comprehensive traits that reflect wheat processing quality. Here, we found that the
dough rheological characteristics were different among three semi-wild wheat varieties (YN, YT, CY) and two cultivated wheat cultivars
(HU and YZ) (Fig. 1A). Therefore, the NAM population composed of the five parents was used to detect QTL of wheat processing
quality. The results show that the NAM population had large variation between three semi-wild RIL populations, CY-RILs, YN-RILs, and
YT-RILs (Fig. 1B). The difference between MlPT and MlPH was not significant, but there were significant correlations between the other dough rheological
characteristics (Fig. 1C). Moreover, one important genetic region, QG-3B.1, associated with all dough rheological characteristics, was
detected (Table 1). In addition, in terms of the association among the four dough rheological characteristics, MlPT was significantly
positively correlated with PkWd and WdEm (Fig. 1C). Specifically, PkWd and WdEm increased with increasing MlPT (verified by QG-
1B.2, QG-1B.4 and QG-3B.3; Fig. 1B-C, Table 1). Although MlPT and MlPH were colocalized between two genetic regions, QG-1B.2 and
QG-1B.4, the correlation between MlPT and MlPH was not significant. We suspect that this may be because MlPT is related to the
protein content, while MlPH is related to the gluten strength and the ability of the dough to resist external forces. WdEm was
significantly positively correlated with PkWd (verified by QG-1B.1) and significantly negatively correlated with MlPH (substantiated by
QG-1B.3, QG-3B.2, and QG-4D, Fig. 1C, Table 1). Eight important genetic regions for wheat processing quality In this study, eight important genetic regions were found on chromosomes 1B (4), 3B (3), and 4D (Table 1). QG-3B.1 was associated
with all dough rheological characteristics, QMlPT-3B.1, QMlPH-3B.3, QPkWd-3B.1, and QWdEm-3B.1, within 4.71 Mb of physical
distance. Three genetic regions, QG-1B.3, QG-3B.2, and QG-4D, influenced MlPH and WdEm. QG-1B.2, located on flank marker
BS00047700_51–IAAV4866, was associated with MlPT, MlPH, and PkWd. QG-1B.4, located on flank marker tplb0048b10_1365–
Ku_c28580_432, influences dough rheological characteristics MlPT, MlPH, and WdEm. In addition, QG-1B.1 and QG-3B.3 were located Page 4/16 Page 4/16 on flank markers wsnp_Ku_rep_c70742_70379526–tplb0059c20_2221 and wsnp_Ex_c64005_62987067–wsnp_BE497740B_Ta_2_1,
respectively, which influence PkWd and WdEm, and PkWd and MlPT, respectively. on flank markers wsnp_Ku_rep_c70742_70379526–tplb0059c20_2221 and wsnp_Ex_c64005_62987067–wsnp_BE497740B_Ta_2_1,
respectively, which influence PkWd and WdEm, and PkWd and MlPT, respectively. on flank markers wsnp_Ku_rep_c70742_70379526–tplb0059c20_2221 and wsnp_Ex_c64005_62987067–wsnp_BE497740B_Ta_2_1,
respectively, which influence PkWd and WdEm, and PkWd and MlPT, respectively. Comparison with previous studies In addition, QWdEm-
2A was previously reported at a similar genetic region [11, 43, 44]. Application potential of semi-wild cultivars in breeding good-quality wheat Page 5/16
Here, a NAM population consisting of three unique semi-wild wheat cultivars of China (CY, YN, and YT), the Chinese domesticated
cultivar YZ, and the British domesticated cultivar HU was used to identify QTL for wheat processing quality. The size of a single RIL Page 5/16 population that constitutes the NAM population limits the precise locations of QTL, but the NAM population based on the
polymorphism between YZ and the other four parents has a series of recombinants containing various recombinant fragments. Based
on the advantages of broader genetic bases and potentially higher mapping resolution, NAM population for comprehensive analysis of
QTLs can achieve repeatability verification of QTLs from public parents across different populations, increase the number of QTL
detected and increase the sensitivity of QTL detection. population that constitutes the NAM population limits the precise locations of QTL, but the NAM population based on the
polymorphism between YZ and the other four parents has a series of recombinants containing various recombinant fragments. Based
on the advantages of broader genetic bases and potentially higher mapping resolution, NAM population for comprehensive analysis of
QTLs can achieve repeatability verification of QTLs from public parents across different populations, increase the number of QTL
detected and increase the sensitivity of QTL detection. Four novel QTL, QMlPH-1B.4, QMlPH-3B.4, QWdEm-1B.2, and QWdEm-3B.2, were stably detected (Table 2). One novel and major QTL,
QMlPH-3B.4, was detected in all four environments and the combined QTL analysis, and its favorable allele came from the good-quality
parent HU. Domesticated parent HU had the longest MlPT, highest MlPH, and widest PkWd and WdEm among the parents (Fig. 1A). QMlPH-3B.4 could be utilized for quality improvement of YZ by increasing MlPT, MlPH, PkWd, and WdEm. We generally think that QTL
detected in multiple environments should also be detectable when using BLUP values. QMlPH-1B.4 was detected in four environments
except for BLUP. This may be because the BLUP value only considers the contribution of genetic factors to the phenotype. Therefore,
we think that environmental effects may explain why that QTL was not detected under BLUP. In summary, to achieve good-quality
wheat production, breeding wheat varieties that contain high-quality genotypes is paramount, but a suitable planting environment is
also necessary [45]. Forty-nine QTL were identified in this study, of which 23 were identified in one environment and combined QTL analysis. Application potential of semi-wild cultivars in breeding good-quality wheat Among the 23
QTL, five, eight, eight and two QTL were for MlPT, MlPH, PkWd, and WdEm, respectively (Table S3). For PkWd, 73% of QTL were
detected in one environment and combined QTL analysis, which could be because PkWd is highly influenced by the environment. This
phenomenon can be verified by the heritability of PkWd, which has the lowest heritability among the four dough rheological
characteristics (Tables S2). Among the 49 QTL, we found that favorable alleles of 17 QTL located on chromosome 1B were provided by
YZ, CY, YN, and YT, while favorable alleles of nine QTL located on chromosome 3B were detected in domesticated parents HU and YZ
(Table S3). Four Chinese cultivars have poor dough characteristics compared with the British domesticated cultivar HU (Fig. 1A). This
phenomenon suggests that there may be genes on chromosome 3B of HU that affect wheat processing quality. The existing gene pool of wheat is narrow because it is composed of current and historical wheat cultivars without allelic variation
from landraces and wild species [23, 33]. The latest research shows that from the perspective of the whole genome level, Tibetan semi-
wild wheat has been de-domesticated from local landraces, and its genome is rich in variation [46]. Therefore, as a valuable resource to
broaden the genetic diversity of wheat breeding, Chinese semi-wild cultivars can be used for genetic research of wheat processing
quality, especially the release of the semi-wild wheat reference genome (Tibetan semi-wild wheat) [46]. The favorable alleles of 31% of
the QTL for wheat processing quality were provided by three semi-wild parents, which may be because semi-wild wheat contains alleles
that are not utilized by existing cultivated wheat varieties [36, 47] (Table S4). Hence, semi-wild wheat varieties can enrich the existing
wheat gene pool, provide broader resources for wheat genetic research, and help in investigating the effect of domestication on the
processing quality of wheat. Conclusions A wheat NAM population consisting of semi-wild and domesticated wheat varieties was used to detect QTL for wheat processing
quality. A total of 49 QTL were identified, of which four novel QTL were stably identified in three or more environments. In addition, 15
of 49 QTL favorable alleles were provided by three semi-wild parents, which indicated that semi-wild wheat contained unique genetic
material that was not used in domesticated varieties. Therefore, semi-wild wheat can be used as a genetic resource to enrich the
existing wheat gene pool and provide more abundant variation for genetic research on wheat processing quality. In addition, the release
of whole genome data of semi-wild wheat (Tibetan semi-wild wheat) [46] provides genomic information for further discovery of
excellent alleles in semi-wild wheat and highlights the significance of studying the role of semi-wild wheat in evolution. Abbreviations NAM, nested association mapping; QTL, quantitative trait loci; RIL, recombinant inbred lines; HMW-GS, high-molecular-weight glutenin
subunits; LMW-GS, low-molecular-weight glutenin subunits; MAS, maker-assisted selection; AACC, American Association of Cereal
Chemists; BLUP, best linear unbiased prediction; ANOVA, analysis of variance; JICIM, joint inclusive composite interval mapping; QC,
QTL cluster; Add, additive effects; MlPT, midline peak time; MlPH, midline peak height; PkWd, peak width; WdEm, width at eight minutes;
YZ, Yanzhan 1; HU, Hussar; YN, Yunnanxiaomai, YT, Yutiandaomai; CY, Chayazheda 29. Plant material and experimental design A wheat NAM population consisting of four RIL populations was constructed to detect QTL for wheat processing quality. Yanzhan 1
(YZ, T. aestivum L.) from Henan Province of the Huanghuai region, China was the common parent. Three of four divergent parents were
semi-wild cultivars in China, including Yunnanxiaomai (YN, T. aestivum ssp. yunnanense King) from Yunnan Province [36],
Yutiandaomai (YT, T. petropavlovskyi Udats. et Migusch.) from Sinkiang, and Cayazheda 29 (CY, T. aestivum ssp. tibetanum Shao) Page 6/16 Page 6/16 from Tibet [37, 46]. The other divergent parent was the British dwarf cultivar Hussar (HU), which is a good-quality cultivar [48]. The four
RIL populations (nine and ten generations of self-pollination) were derived using a single seed descent method. The hybridizations of
YZ with YN, YT, CY, and HU ultimately yielded 98, 93, 82, and 97 lines, respectively. from Tibet [37, 46]. The other divergent parent was the British dwarf cultivar Hussar (HU), which is a good-quality cultivar [48]. The four
RIL populations (nine and ten generations of self-pollination) were derived using a single seed descent method. The hybridizations of
YZ with YN, YT, CY, and HU ultimately yielded 98, 93, 82, and 97 lines, respectively. The NAM population along with the five parents were planted in De’zhou (E1, 116.39°E, 37.38°N), Tai’an (E2, 117.17°E, 36.17°N) and
He’ze (E3, 115.50°E, 35.57°N) in Shandong Province during 2015–2016. The materials were planted again in Tai’an (E4) during 2016–
2017. In each environment, each plot comprised two rows with a 2.0 m row length, 0.25 m row spacing, and 50 seeds per row. Two
replicates were performed under each environment. All fields were managed in accordance with standard local practices. Traits investigated The plants in each plot were harvested to evaluate the wheat processing quality. The moisture (%) and protein (%) of grain and flour of
the NAM population were determined by near-infrared reflectance spectroscopy on a Perten Diode Array 7200 (Perten Instruments,
Huddinge, Sweden) instrument according to the methods of the American Association of Cereal Chemists (AACC) 39-10 and 39-11 [49]. The grains were conditioned to 14% moisture content and then milled using a Quadrumat Junior (Brabender GmbH & Co. KG Duisburg,
Germany.) according to the methods of AACC 26-95 and 26-50. Then, a 10 g-Mixograph (National Mfg. Co. Nebraska, America) was
used to carry out rheological tests according to the AACC 54-40A method. For each of the samples, two planting replications were used
for phenotypic data collection, and the following parameters were recorded: MlPT, midline peak time (min); MlPH, midline peak height
(%); PkWd, peak width (%); and WdEm, width at eight minutes (%). Statistical analysis of phenotypic data The best linear unbiased prediction (BLUP) for each line of wheat processing quality was counted across environments using the “lmer”
function implemented in the R package lme4 (https://cran.r-project.org/web/packages/lme4/index.html). Each BLUP was used to
calculate the pairwise correlations for phenotypic data using the “rcorr” function implemented in the R package Hmisc. Boxplots for
phenotypic data were obtained from the BLUP value using Origin Pro V9.1 software (https://www.originlab.com/). Analysis of variance
(ANOVA) for the five parents in wheat processing quality was calculated by Statistics Program for Social Sciences V20 software. Furthermore, the heritabilities were calculated through the AOV function of IciMapping V4.1 software using the formula h2 = VG / (VG +
VGEI /e + Ve/er), where VG, VGEI and Ve are the variances of G (genotypes), GEI (genotype × environment interactions) and the error,
respectively; e is the number of environments; and r is the number of replications [50]. QTL analysis An integrated high-density linkage map (containing 2009 SNP markers) published previously was used in this study [51], and the
procedure was as follows: first, the redundant markers of the 90,000 SNP array were processed through the “BIN” function of
IciMapping V4.1. Second, the remaining markers were divided into different linkage groups through the “MAP” function of IciMapping
V4.1 [50]. Third, based on the Kosambi mapping function, we constructed four individual maps of the RILs. Finally, four individual
maps were combined with Join Map V4.0 (https://www.kyazma.nl/index.php/JoinMap/). The averaged value for each line in each
environment was used to conduct individual environment QTL analysis, and BLUP values across four environments for each line were
used for combined QTL analysis. QTL detection for wheat processing quality was performed by joint inclusive composite interval
mapping (JICIM) in IciMapping V4.1 software [50]. Using this method, the walking step was set as 1.0 cM, and a stepwise regression
probability of 0.001 was used to identify QTL. A QTL was identified when the LOD score was greater than 2.5 in the NAM population
and greater than 2.0 in at least one RIL population. In this study, QTL clusters affecting quality-related traits (MlPT, MlPH, PkWd, and
WdEm) were defined with the prefix “QC”. Funding The cost of microarray as well as other materials was founded by the National Key Research and Development Program
(2016YFD0100102-2), and the labor cost was founded by the Project of Shandong Province Higher Educational Science and
Technology Program (J16LF06). The funding body had no role in the design of the study and collection, analysis, and interpretation of
data and in writing the manuscript. Authors' contributions YGB and PSS conceived and designed the research. JMH performed the analysis, interpreted the results and drafted the manuscript. GLX, GXZ and XM grew plant material. PJ constructed the linkage map. YZ revised the manuscript. LXX and JY carried out the work of
material planting management. All authors read and approved the final version of the manuscript. Authors' Information 1State Key Laboratory of Crop Biology, Shandong Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural
University, Taian 271018, P.R.China. 2 Lianyungang Academy of Agricultural Sciences, Lianyungang 222000, P.R. China 2 Lianyungang Academy of Agricultural Sciences, Lianyungang 222000, P.R. China 3Yantai Academy of Agricultural Sciences in Shandong Province, Yantai 265500, P.R. China 3Yantai Academy of Agricultural Sciences in Shandong Province, Yantai 265500, P.R. China 4Agricultural Technology Station, Sunwu Sub-district Office, Huimin County, Shandong Province 5College of Agriculture, Liaocheng University, Liaocheng 252059, P.R. China Acknowledgements We thank to Prof. XYK and JZJ for the RIL populations materials. Consent for publish Not applicable. Declarations Page 7/16 Page 7/16 Ethics approval and consent to participate Ethics approval and consent to participate
Not applicable. Not applicable. Competing interests The authors declare that they have no competing interests. Availability of data and materials All data generated or analysed during this study are included in this published article [and its supplementary information files]. Competing interests References 1. Brenchley R, Spannagl M, Pfeifer M, Barker GLA, D’Amore R, Allen AM, McKenzie N, Kramer M, Kerhornou A, Bolser D, Kay S, Waite
D, Trick M, Bancroft I, Gu Y, Huo N, Luo MC, Sehgal S, Gill B, Kianian S, Anderson O, et al. Analysis of the bread wheat genome using
whole-genome shotgun sequencing. Nature. 2012; 491:705–710. 1. Brenchley R, Spannagl M, Pfeifer M, Barker GLA, D’Amore R, Allen AM, McKenzie N, Kramer M, Kerhornou A, Bolser D, Kay S, Waite
D, Trick M, Bancroft I, Gu Y, Huo N, Luo MC, Sehgal S, Gill B, Kianian S, Anderson O, et al. Analysis of the bread wheat genome using
whole-genome shotgun sequencing. Nature. 2012; 491:705–710. 2. Hussain W, Baenziger PS, Belamkar V, Guttieri MJ, Venegas JP, Easterly A, Sallam A, Poland J. Genotyping-by-sequencing derived
high-density linkage map and its application to QTL mapping of flag leaf traits in bread wheat. Sci. Rep. 2017;7:16394. 3. Mann G, Diffey S, Cullis B, Azanza F, Martin D, Kelly A, McIntyre L, Schmidt A, Ma WJ, Nath Z, Kutty I, Leyne PE, Rampling L, Quail
KJ, Morell MK. Genetic control of wheat quality: interactions between chromosomal regions determining protein content and
composition, dough rheology, and sponge and dough baking properties. Theor. Appl. Genet. 2009; 118:1519–1537. 3. Mann G, Diffey S, Cullis B, Azanza F, Martin D, Kelly A, McIntyre L, Schmidt A, Ma WJ, Nath Z, Kutty I, Leyne PE, Rampling L, Quail
KJ, Morell MK. Genetic control of wheat quality: interactions between chromosomal regions determining protein content and
composition, dough rheology, and sponge and dough baking properties. Theor. Appl. Genet. 2009; 118:1519–1537. Page 8/16 Page 8/16 4. Michel S, Kummer C, Gallee M, Hellinger J, Ametz C, Akgol B, Epure D, Gungor H, Loschenberger F, Buerstmayr H. Improving the
baking quality of bread wheat by genomic selection in early generations. Theor. Appl. Genet. 2018;131:477–493. 5. Payne PI, Holt LM, Worland AJ, Law CN. Structural and genetical studies on the high-molecular-weight subunits of wheat glutenin. Theor. Appl. Genet. 1982;63:129–138. 6. Cassidy BG, Dvorak J, Anderson OD. The wheat low-molecular-weight glutenin genes: characterization of six new genes and
progress in understanding gene family structure. Theor. Appl. Genet. 1998;96:743–750. 7. Krystkowiak K, Langner M, Adamski T, Salmanowicz BP, Kaczmarek Z, Krajewski P, Surma M. References Interactions between Glu-1 and Glu-3
loci and associations of selected molecular markers with quality traits in winter wheat (Triticum aestivum L.) DH lines. J. Appl. Genet. 2017;58:37–48. 8. Gao S, Gu Y, Wu J, Coleman-Derr D, Huo N, Crossman C, Jia J, Zuo Q, Ren Z, Anderson OD, Kong X. Rapid evolution and complex
structural organization in genomic regions harboring multiple prolamin genes in the polyploid wheat genome. Plant Mol. Biol. 2007;65:189–203. 9. Deng Z, Fang W, Guo X, Zhao X, Guo H, Hu S, Tian J. Genetic dissection of interactions between wheat flour starch and its
components in two populations using two QTL mapping methods. Mol. Breed. 2018;38:41. 10. Liu T, An Y, Liu K, Wang F, Xie C, Zhang Y, Guan X, Tian J, Chen J. A genetic analysis of the quality of northern-style Chinese
steamed bread. Mol. Breed. 2017;37:41. 11. Liu J, He Z, Wu L, Bai B, Wen W, Xie C, Xia X. Genome-wide linkage mapping of QTL for black point reaction in bread wheat
(Triticum aestivum L.). Theor. Appl. Genet. 2016;129:2179–2190. 12. Cui F, Fan X, Chen M, Zhang N, Zhao C, Zhang W, Han J, Ji J, Zhao X, Yang L, Zhao Z, Tong Y, Wang T, Li J. QTL detection for wheat
kernel size and quality and the responses of these traits to low nitrogen stress. Theor. Appl. Genet. 2016;129:469–484. 13. Zhang Y, Tang J, Zhang Y, Yan J, Xiao Y, Zhang Y, Xia X, He Z. QTL mapping for quantities of protein fractions in bread wheat
(Triticum aestivum L.). Theor. Appl. Genet. 2011;122:971–987. 14. Suprayogi Y, Pozniak CJ, Clarke FR, Clarke JM, Knox RE, Singh AK. Identification and validation of quantitative trait loci for grain
protein concentration in adapted Canadian durum wheat populations. Theor. Appl. Genet. 2009;119:437–448. 15. Zhang W, Chao S, Manthey F, Chicaiza O, Brevis JC, Echenique V, Dubcovsky J. QTL analysis of pasta quality using a composite
microsatellite and SNP map of durum wheat. Theor. Appl. Genet. 2008;117:1361–1377. 16. Arbelbide M, Bernardo R. Mixed-model QTL mapping for kernel hardness and dough strength in bread wheat. Theor. Appl. Genet. 2006;112:885–890. 17. Cabrera A, Guttieri M, Smith N, Souza E, Sturbaum A, Hua D, Griffey C, Barnett M, Murphy P, Ohm H, Uphaus J, Sorrells M, Heffner E,
Brown-Guedira G, Sanford DV, Sneller C. Identification of milling and baking quality QTL in multiple soft wheat mapping
populations. Theor. Appl. Genet. 2015;128:2227–2242. References Genetic properties of the maize nested
association mapping population. Science. 2009;325:737-740. 30. Kidane YG, Gesesse CA, Hailemariam BN, Desta EA, Mengistu DK, Fadda C, Pe ME, Dell-Acqua M. A large nested association
mapping population for breeding and quantitative trait locus mapping in Ethiopian durum wheat. Plant Biotechnol. J. 2019;17:1380–1393. 31. Jordan KW, Shichen W, Fei H, et al. The genetic architecture of genome-wide recombination rate variation in allopolyploid wheat
revealed by nested association mapping. Plant J. 2018;95. 32. Bajgain P, Rouse M, Tsilo TJ, Macharia GK, Bhavani S, Jin Y, Anderson JA, Bai G. Nested Association Mapping of Stem Rust
Resistance in Wheat Using Genotyping by Sequencing. Plos one. 2016;11:e0155760. 33. Pont C, Leroy T, Seidel M, Tondelli A, Duchemin W, Armisen D, Lang D, Bustos-Korts D, Goue N, Balfourier F, Molnar-Lang M, Lage J,
Kilian B, Ozkan H, Waite D, Dyer S, Letellier T, Alaux M, Wheat and Barley Legacy for Breeding Improvement consortium, Russell J,
Keller B, et al. Tracing the ancestry of modern bread wheats. Nat. Genet. 2019;51:905–911. 34. Zhou Y, Zhao X, Li Y. et al. Triticum population sequencing provides insights into wheat adaptation. Nat. Genet. 2020;
https://doi.org/10.1038/s41588-020-00722-w. 35. Wei Y, Zheng Y, Liu D, Zhou Y, Lan X. HMW-glutenin and gliadin variations in Tibetan weedrace, Xinjiang rice wheat and Yunnan
hulled wheat. Genet. Resour. Crop Ev. 2002;49:327–330. 36. Chen F, Yu Y, Xia X, He Z. Prevalence of a novel puroindoline b allele in Yunnan endemic wheats (Triticum aestivum ssp. yunnanense King). Euphytica. 2007;156:39–46. 37. Jiang Y, Wang J, Luo W, Wei Y, Qi P, Liu Y, Jiang Q, Peng Y, Chen G, Dai S, Zheng Y, Lan X. Quantitative trait locus mapping for seed
dormancy in different post-ripening stages in a Tibetan semi-wild wheat (Triticum aestivum ssp. tibetanum Shao). Euphytica. 2015;203:557–567. 38. Liu S, Chao S, Anderson JA. New DNA markers for high molecular weight glutenin subunits in wheat. Theor. Appl. Genet. 2008;118:177–183. 39. He X, Wang J, Ammar K, Peña RJ, Xia X, He Z. Allelic variants at the Psy-A1 and Psy-B1 loci in durum wheat and their associations
with grain yellowness. Crop Sci. 2009;49:2058–2064. 40. 40. Butow BJ, Gale KR, Ikea J, Juhasz A, Bedo Z, Tamas L, Gianibelli MC. Dissemination of the highly expressed Bx7 glutenin
subunit (Glu-B1al allele) in wheat as revealed by novel PCR markers and RP-HPLC. Theor. Appl. Genet. 2004;109:1525–1535. 41. 41. References 18. Deng Z, Tian J, Chen F, Li W, Zheng F, Chen J, Shi C, Sun C, Wang S, Zhang Y. Genetic dissection on wheat flour quality traits in two
related populations. Euphytica. 2015;203:221–235. 19. Lado B, Vazquez D, Quincke M, Silva P, Aguilar I, Gutierrez L. Resource allocation optimization with multi-trait genomic prediction
for bread wheat (Triticum aestivum L.) baking quality. Theor. Appl. Genet. 2018;131:2719–2731. 20. Hamer RJ, MacRitchie F, Weegels PL. Chapter 6: Structure and functional properties of gluten, in: K. Khan, P.R. Shewry (Eds), Wheat:
Chemistry and Technology, AACC International, Inc., St. Paul, Minnesota, USA. 2009:153–178. 21. Hayes BJ, Panozzo J, Walker CK, Choy AL, Kant S, Wong D, Tibbits J, Daetwyler HD, Rochfort S, Hayden MJ, Spangenberg GC. Accelerating wheat breeding for end-use quality with multi-trait genomic predictions incorporating near infrared and nuclear
magnetic resonance-derived phenotypes. Theor. Appl. Genet. 2017;130:2505–2519. 22. Tsilo TJ, Nygard G, Khan K, Simsek S, Hareland GA, Chao S, Anderson JA. Molecular genetic mapping of QTL associated with flour
water absorption and farinograph related traits in bread wheat. Euphytica. 2013;194:293–302. 23. Li Y, Zhou R, Wang J, Liao X, Branlard G, Jia J. Novel and favorable QTL allele clusters for end-use quality revealed by introgression
lines derived from synthetic wheat. Mol. Breed. 2012;29:627–643. 24. Edae EA, Byrne PF, Haley SD, Lopes MS, Reynolds MP. Genome-wide association mapping of yield and yield components of spring
wheat under contrasting moisture regimes. Theor. Appl. Genet. 2014;127:791–807. 24. Edae EA, Byrne PF, Haley SD, Lopes MS, Reynolds MP. Genome-wide association mapping of yield and yield components of spring
wheat under contrasting moisture regimes. Theor. Appl. Genet. 2014;127:791–807. 25. Doerge RW. Multifactorial genetics: mapping and analysis of quantitative trait loci in experimental populations. Nat. Rev. Genet. 2002;3:43–52. 25. Doerge RW. Multifactorial genetics: mapping and analysis of quantitative trait loci in experimental populations. Nat. Rev. Genet. 2002;3:43–52. Page 9/16 Page 9/16 26. Holland JB. Genetic architecture of complex traits in plants. Curr. Opin. Plant Biol. 2007;10:156–161). 27. Jordan K. Sequence-based map development of wheat NAM populations. Plant & Animal Genome. 2015. 28. Tian F, Bradbury PJ, Brown PJ, Hung H, Sun Q, Flint-Garcia S, Rocheford TR, McMullen MD, Holland JB, Buckler ES. Genome-wide
association study of leaf architecture in the maize nested association mapping population. Nat. Genet. 2011; doi:10.1038/ng.746. 29. McMullen MD, Kresovich S, Villeda HS, Bradbury P, Li H, Sun Q, Flint-Garcia S. et al. 2008;117:29–35. 49. AACC International, Approved methods of the American Association of Cereal Chemists, Tenth edition, AACC International. St. Paul,
Minnesota, USA. 2000. 51. Hu J, Wang X, Zhang G, Jiang P, Chen W, Hao Y, Ma X, Xu S, Jia J, Kong L, Wang H. QTL mapping for yield-related traits in wheat
based on four RIL populations. Theor. Appl. Genet. 2020;133:917–933. 51. Hu J, Wang X, Zhang G, Jiang P, Chen W, Hao Y, Ma X, Xu S, Jia J, Kong L, Wang H. QTL mapping for yield-related traits in wheat
based on four RIL populations. Theor. Appl. Genet. 2020;133:917–933. References Carter AH, Garland-Campbell K, Morris CF, Kidwell KK. Chromosomes 3B and 4D are associated with several milling and baking
quality traits in a soft white spring wheat (Triticum aestivum) population. Theor. Appl. Genet. 2012;124:1079–1096. 42. 42. Li J, Cui F, Ding A, Zhao C, Wang X, Wang L, Bao Y, Qi X, Li X, Gao J, Feng D, Wang H. QTL detection of seven quality traits in
wheat using two related recombinant inbred line populations. Euphytica. 2012;183:207–226. 43. 43. Sun D, He Z, Xia X, Zhang L, Morris CF, Appels R, Ma W, Wang H. A novel STS marker for polyphenol oxidase activity in bread
wheat. Mol. Breed. 2005;16:209–218. 44. 44. He X, He Z, Zhang L, Sun D, Morris CF, Fuerst EP, Xia X. Allelic variation of polyphenol oxidase (PPO) genes located on
chromosomes 2A and 2D and development of functional markers for the PPO genes in common wheat. Theor. Appl. Genet. 2007;115:47–58. 45. 45. Asseng S, Martre P, Maiorano A, Rötter RP, O’Leary GJ, Fitzgerald GJ, Girousse C, Motzo R, Giunta F, Babar MA, Reynolds MP,
Kheir AMS, Thorburn PJ, Waha K, Ruane AC, Aggarwal PK, Ahmed M, Balkovič J, Basso B, Biernath C, Bindi M, Cammarano D,
Challinor AJ, De Sanctis G, et al. Climate change impact and adaptation for wheat protein. Glob. Change Biol. 2019;25:155–17. 46. Guo W, Xin M, Wang Z et al. Origin and adaptation to high altitude of Tibetan semi-wild wheat. Nat. Commun. 2020; 11:5085. 46. Guo W, Xin M, Wang Z et al. Origin and adaptation to high altitude of Tibetan semi-wild wheat. Nat. Commun. 2020; 11:5085. 47. Du X, Hu J, Ma X. et al. Molecular characterization and marker development for high molecular weight glutenin subunit 1Dy12** 47. Du X, Hu J, Ma X. et al. Molecular characterization and marker development for high molecular weight glutenin subunit 1Dy12**
from Yunnan hulled wheat. Mol. Breeding 2019;39:4. Page 10/16
48. Wilde F, Schon CC, Korzun V, Ebmeyer E, Schmolke M, Hartl L, Miedaner T. Marker-based introduction of three quantitative-trait loci
conferring resistance to Fusarium head blight into an independent elite winter wheat breeding population. Theor. Appl. Genet. Page 10/16 2008;117:29–35. Tables Table 1 Eight important genetic regions associated with multiple dough rheological characteristics. Genetic
region
QTL
Flanking markers of peak
QG-1B.1
QPkWd-1B.1, QWdEm-1B.1
wsnp_Ku_rep_c70742_70379526
tplb0059c20_2221
QG-1B.2
QMlPT-1B.1, QMlPH-1B.2, QPkWd-1B.2
BS00047700_51
IAAV4866
QG-1B.3
QWdEm-1B.2, QMlPH-1B.3
Tdurum_contig20299_368
BobWhite_c48550_198
QG-1B.4
QMlPT-1B.2, QWdEm-1B.3, QMlPH-1B.4
tplb0048b10_1365
Ku_c28580_432
QG-3B.1
QMlPT-3B.1, QPkWd-3B.1, QWdEm-3B.1,
QMlPH-3B.3
wsnp_Ex_c3257_6003626
wsnp_Ex_c8715_14590273
QG-3B.2
QWdEm-3B.2, QMlPH-3B.4
wsnp_Ku_c29102_39008953
wsnp_Ex_c64005_62986957
QG-3B.3
QMlPT-3B.2, QPkWd-3B.2
wsnp_Ex_c64005_62987067
wsnp_BE497740B_Ta_2_1
QG-4D
QMlPH-4D, QWdEm-4D
BS00103682_51
Ex_c42133_630
MlPT, midline peak time; MlPH, midline peak height; PkWd, peak width; WdEm, width at eight minutes; QG, QTL cluster. able 1 Eight important genetic regions associated with multiple dough rheological characteristics , midline peak height; PkWd, peak width; WdEm, width at eight minutes; QG, QTL cluster. MlPT, midline peak time; MlPH, midline peak height; PkWd, peak width; WdEm, width at eight minutes; QG, QTL cluster. Table 2
Four novel QTL that were stably identified in three or more environments. QTL
Environment
Position
Flank markers of peak
LOD
PVE (%)
QMlPH-
1B.4
E1/E2/E3/E4
44
tplb0048b10_1365
Ku_c28580_432
4.0129–
9.7453
1.4168–
4.503
QMlPH-
3B.4
E1/E2/E3/E4/BLUP
63
wsnp_Ku_c29102_39008953
wsnp_Ex_c64005_62986957
6.365–
32.0377
1.711–
10.8244
QWdEm-
1B.2
E1/E2/E3/BLUP
38
Tdurum_contig20299_368
BobWhite_c48550_198
4.6957–
5.7802
0.5107–
2.4201
QWdEm-
3B.2
E1/E2/E3
63
wsnp_Ku_c29102_39008953
wsnp_Ex_c64005_62986957
6.1816–
20.4799
2.5598–
9.7032
MlPT, midline peak time; MlPH, midline peak height; PkWd, peak width; WdEm, width at eight minutes. E1, 2016 De’zhou; E2, 2016
Tai’an; E3, 2016 He’ze; E4, 2017 Tai’an; BLUP, best linear unbiased prediction. PVE, phenotypic variance explained. 49. AACC International, Approved methods of the American Association of Cereal Chemists, Tenth edition, AACC International. St. Paul,
Minnesota, USA. 2000. Figures Page 11/16 Page 11/16 Page 11/16 Figure 1
Phenotypic data of wheat processing quality of the five parents and the NAM population. A. The four dough rheological characteristi
of the five parents (a–e) and the ANOVA between five parents of four dough rheological characteristics (f–i). Labels A and B indicate
significant differences at the level of P < 0.01, and labels a and b indicate significant differences at the level of P < 0.05. B. The boxp
for four dough rheological characteristics of four RIL populations. The different color lines of black, green, yellow, red, and blue indica
the five parents of the NAM population YZ, CY, HU, YN, and YT, respectively. C. The relationships between four dough rheological
characteristics of the NAM population. YZ, Yanzhan 1; CY, Chayazheda 29; HU, Hussar; YN, Yunnanxiaomai; YT, Yutiandaomai. MlPT,
midline peak time; MlPH, midline peak height; PkWd, peak width; WdEm, width at eight minutes. Figure 1 Phenotypic data of wheat processing quality of the five parents and the NAM population. A. The four dough rheological characteristics
of the five parents (a–e) and the ANOVA between five parents of four dough rheological characteristics (f–i). Labels A and B indicate
significant differences at the level of P < 0.01, and labels a and b indicate significant differences at the level of P < 0.05. B. The boxplot
for four dough rheological characteristics of four RIL populations. The different color lines of black, green, yellow, red, and blue indicate
the five parents of the NAM population YZ, CY, HU, YN, and YT, respectively. C. The relationships between four dough rheological
characteristics of the NAM population. YZ, Yanzhan 1; CY, Chayazheda 29; HU, Hussar; YN, Yunnanxiaomai; YT, Yutiandaomai. MlPT,
midline peak time; MlPH, midline peak height; PkWd, peak width; WdEm, width at eight minutes. Page 12/16 Page 12/16 Figure 1
Phenotypic data of wheat processing quality of the five parents and the NAM population. A. The four dough rheological characteristi
of the five parents (a–e) and the ANOVA between five parents of four dough rheological characteristics (f–i). Labels A and B indicate
significant differences at the level of P < 0.01, and labels a and b indicate significant differences at the level of P < 0.05. B. The boxp
for four dough rheological characteristics of four RIL populations. The different color lines of black, green, yellow, red, and blue indica
the five parents of the NAM population YZ, CY, HU, YN, and YT, respectively. C. The relationships between four dough rheological
characteristics of the NAM population. YZ, Yanzhan 1; CY, Chayazheda 29; HU, Hussar; YN, Yunnanxiaomai; YT, Yutiandaomai. MlPT,
midline peak time; MlPH, midline peak height; PkWd, peak width; WdEm, width at eight minutes. Figure 2 Identified QTL for wheat processing quality detected in the NAM population. Red, blue, green and purple indicate the midline peak time,
the midline peak height, the peak width, and the width at eight minutes, respectively. The outermost part of the graph represents the
QTL/gene reported in the previous study. Figure 1 Phenotypic data of wheat processing quality of the five parents and the NAM population. A. The four dough rheological characteristics
of the five parents (a–e) and the ANOVA between five parents of four dough rheological characteristics (f–i). Labels A and B indicate
significant differences at the level of P < 0.01, and labels a and b indicate significant differences at the level of P < 0.05. B. The boxplot
for four dough rheological characteristics of four RIL populations. The different color lines of black, green, yellow, red, and blue indicate
the five parents of the NAM population YZ, CY, HU, YN, and YT, respectively. C. The relationships between four dough rheological
characteristics of the NAM population. YZ, Yanzhan 1; CY, Chayazheda 29; HU, Hussar; YN, Yunnanxiaomai; YT, Yutiandaomai. MlPT,
midline peak time; MlPH, midline peak height; PkWd, peak width; WdEm, width at eight minutes. Page 13/16 Figure 2
Identified QTL for wheat processing quality detected in the NAM population. Red, blue, green and purple indicate the midline peak time,
the midline peak height, the peak width, and the width at eight minutes, respectively. The outermost part of the graph represents the
QTL/gene reported in the previous study. Figure 2 Identified QTL for wheat processing quality detected in the NAM population. Red, blue, green and purple indicate the midline peak time,
the midline peak height, the peak width, and the width at eight minutes, respectively. The outermost part of the graph represents the
QTL/gene reported in the previous study. Identified QTL for wheat processing quality detected in the NAM population. Red, blue, green and purple indicate the midline peak time,
the midline peak height, the peak width, and the width at eight minutes, respectively. The outermost part of the graph represents the
QTL/gene reported in the previous study. Page 14/16 Page 14/16 Supplementary Files This is a list of supplementary files associated with this preprint. Click to download. Additionalfiles.docx Additionalfiles.docx
Additionalfiles.docx
Supplementarytable1.xlsx
Supplementarytable1.xlsx
Supplementarytable2.xlsx
Supplementarytable2.xlsx
Supplementarytable3.xlsx Additionalfiles.docx Supplementarytable1.xlsx Supplementarytable1.xlsx Supplementarytable2.xlsx Supplementarytable2.xlsx Supplementarytable3.xlsx Page 15/16 Page 15/16 Supplementarytable3.xlsx
Supplementarytable4.xlsx
Supplementarytable4.xlsx Supplementarytable3.xlsx Supplementarytable4.xlsx Page 15/16 Supplementarytable3.xlsx
Supplementarytable4.xlsx
Supplementarytable4.xlsx Page 16/16
|
https://openalex.org/W4233868636
|
https://europepmc.org/articles/pmc8447757?pdf=render
|
English
| null |
Study TPX-100-5: Intra-Articular TPX-100 Significantly Delays Pathological Bone Shape Change and Stabilizes Cartilage in Moderate to Severe Bilateral Knee OA
|
Research Square (Research Square)
| 2,021
|
cc-by
| 6,783
|
© The Author(s). 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License,
which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give
appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if
changes were made. The images or other third party material in this article are included in the article's Creative Commons
licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons
licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain
permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the
data made available in this article, unless otherwise stated in a credit line to the data. Study TPX-100-5: intra-articular TPX-100
significantly delays pathological bone
shape change and stabilizes cartilage in
moderate to severe bilateral knee OA Dawn McGuire1*
, Michael Bowes2, Alan Brett2, Neil A. Segal3, Meghan Miller1, David Rosen1 and
Yoshinari Kumagai1 Abstract Registered 15 August 2013 l registration: NIH ClinicalTrials.gov, NCT01925261. Registered 15 August 2013 Keywords: B-score, Bone shape, DMOAD, Machine learning, Osteoarthritis, TPX-100 * Correspondence: dawn.mcguire@orthotrophix.com p
g
@
1OrthoTrophix, Foster City, CA 94404, USA
Full list of author information is available at p
g
@
1OrthoTrophix, Foster City, CA 94404, USA McGuire et al. Arthritis Research & Therapy (2021) 23:242
https://doi.org/10.1186/s13075-021-02622-8 McGuire et al. Arthritis Research & Therapy (2021) 23:242
https://doi.org/10.1186/s13075-021-02622-8 (2021) 23:242 McGuire et al. Arthritis Research & Therapy
https://doi.org/10.1186/s13075-021-02622-8 p
g
p
1OrthoTrophix, Foster City, CA 94404, USA
Full list of author information is available at the end of the article Abstract Background: TPX-100, a promotor of osteoblast and chondroblast differentiation, is a potential osteoarthritis (OA)
therapy. This retrospective study compared MRI 3D femoral bone shape changes (B-scores) after intra-articular TPX-
100 or placebo and analyzed the relationship between cartilage thickness and bone shape change over 12 months. Methods: One hundred and four participants with bilateral moderate to severe knee cartilage defects were
randomized to receive TPX-100 (200 mg) or placebo. Each subject’s contralateral placebo-treated knee served as a
paired internal control. After MRI quality control, 78/93 subjects (84%; 156 knees) were analyzed for quantitative
femoral B-score and cartilage thickness. All analyses were performed centrally, blind to treatment assignment and
clinical data. Results: TPX-100-treated knees (n = 78) demonstrated a statistically significant decrease in pathologic bone shape
change compared with placebo-treated knees at 6 and 12 months: 0.0298 (95% C.I. −0.037, 0.097) vs 0.1246 (95%
C.I. 0.067, 0.182) (P = 0.02), and 0.0856 (95% C.I. 0.013, 0.158) vs. 0.1969 (95% C.I. 0.123, 0.271) (P = 0.01), respectively. The correlation between bone shape change and medial and total tibiofemoral cartilage thickness changes at 12
months was statistically significant in TPX-100-treated knees (P < 0.01). Results: TPX-100-treated knees (n = 78) demonstrated a statistically significant decrease in pathologic bone shape
change compared with placebo-treated knees at 6 and 12 months: 0.0298 (95% C.I. −0.037, 0.097) vs 0.1246 (95%
C.I. 0.067, 0.182) (P = 0.02), and 0.0856 (95% C.I. 0.013, 0.158) vs. 0.1969 (95% C.I. 0.123, 0.271) (P = 0.01), respectively. The correlation between bone shape change and medial and total tibiofemoral cartilage thickness changes at 12
months was statistically significant in TPX-100-treated knees (P < 0.01). Conclusions: This is the first report of a potential therapy demonstrating a significant effect on bone shape
measured by B-score in knee OA. These data, in combination with previously reported statistically significant and
clinically meaningful improvements in WOMAC physical function versus placebo, support TPX-100 as a candidate
for disease modification in knee OA. Conclusions: This is the first report of a potential therapy demonstrating a significant effect on bone shape
measured by B-score in knee OA. These data, in combination with previously reported statistically significant and
clinically meaningful improvements in WOMAC physical function versus placebo, support TPX-100 as a candidate
for disease modification in knee OA. Trial registration: NIH ClinicalTrials.gov, NCT01925261. Registered 15 August 2013 Trial registration: NIH ClinicalTrials.gov, NCT01925261. RESEARCH ARTICLE Open Access Background measures. Subjects were screened clinically and with
MRI, inclusion criteria were applied for patellofemoral
osteoarthritis (PFOA) severity (grades 2–3), and ag-
nostic for osteoarthritis (OA) severity in other joint
compartments. Synovitis and meniscal damage were
among the exclusion criteria for enrollment. The pre-
selected primary efficacy outcome measure in this trial
was the 6-month change in patellar cartilage thickness as
measured using standardized magnetic resonance imaging
(MRI) in TPX-100-treated knees compared with placebo-
exposed knees (see Fig. 1; CONSORT diagram). Briefly,
part A of the study evaluated safety of 4 once-weekly IA
doses of TPX-100 in sequential dose cohorts (25, 50, 100,
and 200 mg/injection) of 6–9 subjects, with progression to
the next dose following Safety Review Committee ap-
proval. All doses were reasonably safe and well tolerated,
and the highest dose, 200 mg/injection, was selected for
part B of the study for evaluation of efficacy and safety. Of
the 118 subjects enrolled for parts A and B, 93 subjects re-
ceived 4 injections of 200 mg TPX-100 and had baseline
MRIs with at least one follow-up scan. Per the statistical
analysis plan, these subjects made up the primary analysis
population. Efficacy outcome measures included MRI car-
tilage measures (6 and 12 months) and patient-reported
outcomes (WOMAC, KOOS, and NRS for pain at 3, 6,
and 12 months). The primary efficacy outcome measure g
TPX-100 is a 23-amino acid peptide derived from
Matrix Extracellular Phosphoglycoprotein (MEPE), a
small
integrin-binding
ligand
N-linked
glycoprotein
(SIBLING) family member. MEPE is highly expressed by
osteocytes, is downregulated in osteoarthritis, and may
play a role in osteoarthritic bone remodeling [1–3]. TPX-100 has been shown to induce articular cartilage
formation in vitro and in vivo. Pre-clinically, TPX-100
(AC-100) was administered by 4 weekly intra-articular
(IA) injections in a standardized full-thickness chondral
defect goat model. After 6 months, TPX-100 demon-
strated robust articular (hyaline) cartilage formation and
increased Type II collagen compared with vehicle-
treated controls as demonstrated by increased type II
collagen (immunostaining) [4]. In humans, a phase II randomized, double-blind,
placebo-controlled,
12-month
trial
of
TPX-100
(NCT01925261)
has
been
completed
to
evaluate
safety, tolerability, and efficacy of IA TPX-100 in sub-
jects with bilateral (ICRS grades 2–3) patellofemoral
(PF) cartilage defects, with or without tibiofemoral
cartilage defects [5, 6]. © The Author(s). 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License,
which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give
appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if
changes were made. The images or other third party material in this article are included in the article's Creative Commons
licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons
licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain
permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the
data made available in this article, unless otherwise stated in a credit line to the data. McGuire et al. Arthritis Research & Therapy (2021) 23:242 Page 2 of 9 McGuire et al. Arthritis Research & Therapy (2021) 23:242 Page 2 of 9 Background Each subject’s contralateral
placebo-treated knee served as a paired internal con-
trol,
intended
to
control
for
effects
of
age,
sex,
weight, genetic factors, and activity levels on outcome Fig. 1 CONSORT diagram for the TPX-100-1 and TPX-100-5 studies Fig. 1 CONSORT diagram for the TPX-100-1 and TPX-100-5 studies McGuire et al. Arthritis Research & Therapy (2021) 23:242 McGuire et al. Arthritis Research & Therapy (2021) 23:242 Page 3 of 9 Page 3 of 9 was patellofemoral cartilage change compared to baseline
at six months in TPX-100 treated versus control knees. There were no significant treatment differences in the pri-
mary efficacy outcome measure at either 6 or 12 months. Semi-quantitative MRI analysis by central readers blind to
clinical data and treatment assignment demonstrated that
only 14% of knees had measurable patellar cartilage thick-
ness changes, limiting study power for this outcome. MRI-based MOAKS evaluation, including cartilage de-
fects, meniscal pathology, and Hoffa’s synovitis, did not
show differences at baseline or follow-up between TPX-
100-treated knees and placebo-exposed knees, and there
were no significant within-knee changes in bone marrow
lesions at 6 or 12 months. In contrast, patient benefit,
measured by WOMAC and KOOS scores, was statistically
significant and clinically meaningful in favor of TPX-100-
treated knees at 6 months compared with placebo, with
robust functional benefits sustained through the end of
the study at 12 months [3]. Post hoc analyses revealed that
68 (73%) of subjects had, in addition to the bilateral patel-
lofemoral cartilage defects for which they were enrolled,
moderate to severe (ICRS 2–4) bilateral tibiofemoral car-
tilage defects. In these subjects, sustained, statistically sig-
nificant, and clinically meaningful clinical benefits in favor
of TPX-100-treated knees were observed, nearly identical
to those in the whole population [6]. outcomes that were very similar in terms of predictive
validity to those using categorical Kellgren-Lawrence
grading (KLG), the conventional radiographic standard
for OA diagnosis. These data provide construct validity
for
this
new,
continuous
scalar
measurement. In
addition, Bowes et al. showed that bone shape is directly
associated with the risk of clinical outcome measures
such as knee pain, functional deficit, and joint failure, as
indicated by total joint replacement, with only small ef-
fect sizes from adjusting models for potential covariates
such as age, sex, ethnicity, body mass index (BMI), align-
ment,
previous
knee
surgery,
non-steroidal
anti-
inflammatory drug (NSAID) use, and smoking status
[11]. Methods Subjects between the ages of 25 and 75 years with bilat-
eral patellofemoral (PF) knee OA were enrolled at 15
sites across the United States. Written informed consent
was obtained prior to study enrollment. All subjects
were screened by MRI, centrally read, to confirm struc-
tural inclusion criteria: bilateral PF cartilage defects
(ICRS grade 2–3) with intact menisci. One knee was
randomly assigned to receive 4 weekly injections of
TPX-100, while the contralateral knee received placebo
(saline) injections that were identical in appearance and
viscosity. Investigators, subjects, sites, and sponsor were
blinded to treatment assignment. The present study (TPX-100-5) was designed as a
retrospective MRI study to investigate femoral bone
shape change at 6 and 12 months after TPX-100 or pla-
cebo administration and to analyze relationships be-
tween
cartilage
thickness
and
femoral
bone
shape
change at 6 and 12 months after TPX-100 or placebo
administration. MRI images were acquired at baseline, 6 and 12
months with 1.5-T clinical MRI scanners using a sagittal
T1-weighted 3D SPGR, FLASH, or FFE sequence with
fat saturation or water excitation. An identical scanner
and knee coil were used for baseline and follow-up mea-
surements of each participant. Acquisition parameters
were as follows: contiguous slice thickness 1.5 mm, in-
plane resolution of 0.31 mm, repetition time 17 ms, echo
time 7 ms, and flip angle 15°. Identical acquisition pa-
rameters were used at baseline and follow-up. Bone shape change, measured by MRI, has been
shown to predict radiographic onset of OA [7], is associ-
ated with radiographic structural progression [8], dis-
criminates people with knee OA from those without
knee OA [9], and is more responsive to change over
time than is radiographic assessment [10]. In each of
these studies, the femur (defined as the whole of the lat-
eral and medial femoral condyles) had greater discrimin-
ation and responsiveness to change than did the tibia or
patella. The femoral bone shape (“B-score”) metric is a
form of statistical z-score that represents the position of
a femoral bone shape along a shape vector from a non-
OA knee shape (origin) toward an OA knee shape (posi-
tive direction). Non-OA and OA knees used to define
this 3-dimensional (3D) shape vector were categorized
using centrally read and adjudicated Kellgren-Lawrence
grading [11]. Background These findings support femoral B-score as a struc-
tural endpoint in clinical trials of disease-modifying
osteoarthritis
drugs
(DMOADs)
and
prompted
the
present analysis. Methods In the present study TPX-100-5, MRIs from all sub-
jects (n = 93) who received, as randomly assigned, 4
weekly injections of TPX-100 (200 mg/injection) in one
knee (index) and placebo in the contralateral knee (con-
trol) with at least one follow-up MRI were eligible for in-
clusion. Image quality was assessed centrally and blind
to clinical data and treatment assignment. Quality was
sufficient to include bilateral knee images from 78 of the
93 subjects (84%) analyzed in study TPX-100-1 (see Fig. 1; CONSORT diagram). Bowes et al. demonstrated in a large observational co-
hort of over 4500 subjects’ knees from the Osteoarthritis
Initiative (OAI) that MRI-measured B-score produced
logistic
regression
models
for
clinically
important Cartilage measurement was performed centrally, blind
to treatment assignment and clinical data, at a single
center (Chondrometrics GmbH, Freilassing, Germany). McGuire et al. Arthritis Research & Therapy (2021) 23:242 Page 4 of 9 Page 4 of 9 McGuire et al. Arthritis Research & Therapy (2021) 23:242 The subchondral bone and cartilage surface areas of the
medial and lateral tibia and medial and lateral weight-
bearing central femoral condyles were traced manually, ex-
cluding cartilage cover of osteophytes [12]. Quantitative
measures of cartilage, including mean cartilage thickness
averaged over the total area of subchondral bone, were
computed using Chondrometrics software [13]. Mean car-
tilage thickness over the total area of subchondral bone for
the medial tibiofemoral compartment was computed by
summing the values of the medial tibia and those of the
weight-bearing medial femoral condyle, and for the lateral
tibiofemoral compartment by summing the values of the
lateral tibia and those of the lateral weight-bearing femoral
condyle. Definitions of the tibiofemoral subregions as well
as measurement reliability (test-retest reproducibility with
repositioning of the joint between acquisitions) have been
published previously [13]. B-score changes over 6 and 12 months were graphic-
ally compared with the 12-month trajectories of “non-
progressor” and “progressor” B-score groups from OAI
data. The non-progressor group was defined as all of
those who did not change by more than 95% of the
smallest detectable difference (SDD) using the slope of
change measured over 4 years. The average slope over
this 4-year period was about 0.04 B-score units per
annum. The progressor group was defined as all of those
who did change by more than 95% of the SDD using the
slope of change measured over slope over 4 years. Methods The
average slope over this 4-year period was about 0.24 B-
score units per annum. SDD was calculated as the 95%
limit of agreement between the first and second image
measurements from test-retest data, using the Bland-
Altman method [11]. In study TPX-100-5, paired Student’s t-tests were used
to compare B-score change from baseline between the
knee receiving IA placebo (control) or TPX-100 (index)
at 6 months and 12 months. For each knee (index, con-
trol) at 6 months and 12 months, the Pearson coeffi-
cients of the changes from baseline between the femur
B-score and cartilage thickness of tibiofemoral variables
were estimated. The two-sided p-values and 95% confi-
dence intervals from the test of the null hypothesis (that
the true correlation coefficient is equal to zero) were
also computed. Statistical significance was set at P <
0.05. The data analysis for this paper was generated
using SAS software version 9. Femur bone surfaces were automatically segmented
using active appearance models (AAMs) at a single cen-
ter (Imorphics Ltd, Manchester, UK). An AAM is a type
of statistical shape model trained using machine learning
to search images. AAMs have proven to be a successful
supervised machine learning method that can produce a
segmented knee bone surface with sub-millimeter accur-
acy [9, 14]. AAMs were constructed using a training set
of 96 knee DESS sequence MRIs that were selected to
provide examples of OA across KLG grades from sub-
jects enrolled in the OAI [15]. The performance of the
AAM has been tested with various other MRI sequences
and slice thicknesses. Mean automated segmentation
accuracy using MRI FLASH sequences is −0.0009
mm, with ± 95th percentiles of error of + 0.34/−0.43
mm, where a positive error represents the automated
surface being outside the reference surface [16]. The
construction of an AAM produces a “shape space,”
spanned by the set of principal components used to
describe the training set of examples. Within this
shape space, an “OA vector” is defined as the line
that passes through the mean shape of two popula-
tions: a population with OA (OA group, defined as
all knees with KLG ≥2 over 4 years of follow-up)
and a population without OA (non-OA group, defined
as knees with KLG of 0 in the same period). Methods Dis-
tances along the femur OA vector are termed “B-
score”, with the origin (B-score = 0) defined as the
mean shape of the non-OA group for each sex, and 1
unit defined as 1 standard deviation of the non-OA
group along the OA vector (positive values toward the
OA Group). Each parameterized femur bone shape was
projected orthogonally onto the OA vector to specify the
corresponding B-score value [11]. Representative exam-
ples of differences in femur bone shape at various B-scores
and a heat map of the areas that change most with in-
creasing B-score are shown in Fig. 2. Results Characteristics of the cohorts from Study TPX-100-1
and the present Study TPX-100-5 were as follows: All per protocol (PPT) subjects: TPX-100-1, N = 93
Mean age, 58.4 (95% C.I. 56.4, 60.4)
Sex, 38 males, 55 females (59.1% females)
Mean BMI, 30.4 (95% C.I. 29.1, 31.7)
Evaluable B-score analyzed cohort: TPX-100-5, N =
78
Mean age, 58.4 (95% C.I. 56.2, 60.6)
Sex, 30 males, 48 females (61.5% Females)
Mean BMI, 30.9 (95% C.I. 29.5, 32.3) Relative proportions of tibiofemoral ICRS grades of
index and control knees of the subjects, determined as
the maximum degree of cartilage defect observed in the
tibiofemoral knee compartment in study TPX-100-1 and
the current study, TPX-100-5, are provided in Table 1. Proportions of B-scores of index and control knees of
the subjects analyzed in TPX-100-5 study are provided
in Table 2. Comparison of the 78 evaluable TPX-100-treated
knees demonstrated a statistically significant decrease in McGuire et al. Arthritis Research & Therapy (2021) 23:242 Page 5 of 9 Fig. 2 Change in shape for the anterior femur (top row) and posterior femur (bottom row), for various B-scores. Red indicates where there is an
increase in size (locally calculated, based on anatomically corresponded triangles from the shape model), blue indicates decrease in size (locally);
scale shows percentage in area size change of each triangle. Change tends to be greatest around the edge of the cartilage plate (osteophyte
region), but it also occurs in central subchondral regions where the bone flattens out. Adapted from Bowes et al. [11] Fig. 2 Change in shape for the anterior femur (top row) and posterior femur (bottom row), for various B-scores. Red indicates where there is an
increase in size (locally calculated, based on anatomically corresponded triangles from the shape model), blue indicates decrease in size (locally);
scale shows percentage in area size change of each triangle. Change tends to be greatest around the edge of the cartilage plate (osteophyte
region), but it also occurs in central subchondral regions where the bone flattens out. Adapted from Bowes et al. [11] B-score change in the femur compared with placebo-
exposed knees at 6 months, 0.0298 (95% C.I. −0.037,
0.097) vs 0.1246 (95% C.I. 0.067, 0.182) (P = 0.02), and
at 12 months, 0.0856 (95% C.I. 0.013, 0.158) vs 0.1969
(95% C.I. 0.123, 0.271) (P = 0.01). Graphical comparisons exposed knees at 6 months, 0.0298 (95% C.I. Results −0.037,
0.097) vs 0.1246 (95% C.I. 0.067, 0.182) (P = 0.02), and
at 12 months, 0.0856 (95% C.I. 0.013, 0.158) vs 0.1969
(95% C.I. 0.123, 0.271) (P = 0.01). Graphical comparisons
Table 1 ICRS grades of index and control knees of the subjects
analyzed in TPX-100-1 and TPX-100-5 studies, respectively at
baseline. ICRS grade in each knee was determined as the
maximum degree of cartilage defect observed in tibiofemoral
knee compartment. Subjects were screened clinically and with
MRI, with inclusion criteria applied for patellofemoral OA
severity (grades 2–3) only, and agnostic for OA severity in other
joint compartments. One control knee failed ICRS grade
measurement
ICRS
grade
TPX-100-1 (N = 93)
TPX-100-5 (N = 78)
Index
Control
Index
Control
4
37% (34)
37% (34)
35% (27)
34% (26)
3
25% (23)
21% (19)
27% (21)
19% (15)
2
19% (18)
21% (19)
18% (14)
22% (17)
1
0% (0)
0% (0)
0% (0)
0% (0)
0
19% (18)
21% (20)
20% (16)
25% (19)
Total
100% (93)
100% (92)
100% (78)
100% (77)
Table 2 Femoral B-scores of index and control knees of the
subjects analyzed in TPX-100-5 study at baseline. Mean B-scores
of index and control knees at baseline were the same. A B-score
of 1.48 corresponds to the mean of the range for a KLG of 2
[11]
Baseline
TPX-100-5 (N = 78)
B-score
Index
Control
< −2 to −1
4% (3)
1% (1)
< −1 to 0
15% (12)
19% (15)
> 0 to 1
24% (19)
24% (19)
> 1 to 2
23% (18)
23% (18)
> 2 to 3
12% (9)
10% (8)
> 3 to 4
15% (12)
10% (8)
> 4 to 5
0% (0)
9% (7)
> 5 to 6
3% (2)
1% (1)
> 6 to 7
1% (1)
1% (1)
> 7 to 8
3% (2)
0% (0)
Total
100% (78)
100% (78)
Mean
1.48
1.48 Table 2 Femoral B-scores of index and control knees of the
subjects analyzed in TPX-100-5 study at baseline. Mean B-scores
of index and control knees at baseline were the same. Results A B-score
of 1.48 corresponds to the mean of the range for a KLG of 2
[11]
Baseline
TPX-100-5 (N = 78)
B-score
Index
Control
< −2 to −1
4% (3)
1% (1)
< −1 to 0
15% (12)
19% (15)
> 0 to 1
24% (19)
24% (19)
> 1 to 2
23% (18)
23% (18)
> 2 to 3
12% (9)
10% (8)
> 3 to 4
15% (12)
10% (8)
> 4 to 5
0% (0)
9% (7)
> 5 to 6
3% (2)
1% (1)
> 6 to 7
1% (1)
1% (1)
> 7 to 8
3% (2)
0% (0)
Total
100% (78)
100% (78)
Mean
1.48
1.48 Table 1 ICRS grades of index and control knees of the subjects
analyzed in TPX-100-1 and TPX-100-5 studies, respectively at
baseline. ICRS grade in each knee was determined as the
maximum degree of cartilage defect observed in tibiofemoral
knee compartment. Subjects were screened clinically and with
MRI, with inclusion criteria applied for patellofemoral OA
severity (grades 2–3) only, and agnostic for OA severity in other
joint compartments. One control knee failed ICRS grade
measurement measurement
ICRS
grade
TPX-100-1 (N = 93)
TPX-100-5 (N = 78)
Index
Control
Index
Control
4
37% (34)
37% (34)
35% (27)
34% (26)
3
25% (23)
21% (19)
27% (21)
19% (15)
2
19% (18)
21% (19)
18% (14)
22% (17)
1
0% (0)
0% (0)
0% (0)
0% (0)
0
19% (18)
21% (20)
20% (16)
25% (19)
Total
100% (93)
100% (92)
100% (78)
100% (77) McGuire et al. Arthritis Research & Therapy (2021) 23:242 Page 6 of 9 Fig. 3 Femoral bone shape changes in TPX-100-treated knees and placebo-exposed knees in bilateral OA subjects and their comparison with
historical bone shape change from the Osteoarthritis Initiative (OAI). All error bars represent standard errors of the mean Fig. 3 Femoral bone shape changes in TPX-100-treated knees and placebo-exposed knees in bilateral OA subjects and t
historical bone shape change from the Osteoarthritis Initiative (OAI). Results All error bars represent standard errors of the mean In TPX-100-treated knees, correlation analyses re-
vealed a statistically significant association between B-
score change and medial femoral condyle cartilage thick-
ness change at 6 months and a statistically significant as-
sociation
between
B-score
change
and
cartilage
thickness change in the entire femoral condyle and en-
tire tibiofemoral (TF) compartments at 12 months (P <
0.05; Table 4). There was also a significant association
between B-score change and cartilage thickness change
in the medial femoral condyle and medial TF compart-
ments at 12 months (P < 0.005; Table 4). of B-score change at 6 months with progressor and non-
progressor groups from OAI data showed B-score trajec-
tories in TPX-100-treated knees that were strikingly
similar to those of OA non-progressors, and, among
placebo-exposed knees, B-score trajectories that were
similar to OA progressors (Fig. 3). The cohort including
all knees, regardless of baseline ICRS grade, demon-
strated B-score changes significantly different in favor of
TPX-100 treatment compared with controls at both 6 (p
= 0.02) and 12 months (p = 0.01). Knees more severely
affected (ICRS grades 3–4) were significantly different in
favor of TPX-100 at 6 months (p = 0.01), with a non-
significant trend (= 0.09) at 12 months (Table 3). Cartilage thickness change in the lateral femoral con-
dyle and lateral TF compartments did not show an asso-
ciation with B-score changes at either 6 or 12 months. There was little or no association between B-score
change and cartilage thickness change in control knees,
with the exception of the medial femoral condyle at 12
months (p = 0.03) (Table 5). Analysis of quartiles of index knee B-score change at
12 months showed a statistically significant association
between less cartilage thickness loss, particularly in the
medial femoral condyle, and lower positive (worsening)
B-score change (Fig. 4). Table 3 Reduction of pathological bone shape change (B-score) by TPX-100 treatment tends to improve in the knees with higher
ICRS grades at baseline. 6 and 12-month B-score changes of index and control knees were compared among the baseline
tibiofemoral ICRS grade cohorts. Data of baseline ICRS grade ≥0 (i.e., all knees) are the same as those shown in Fig. 3 and
comparison between index and control knees was by paired t-test. Results Because index and control knees in the higher baseline ICRS
grade cohorts did not necessarily belong to the same subjects, the comparisons in these three cohorts are by unpaired t-tests Baseline
tibiofemoral
ICRS grade
Mean baseline B-score
(number of knees)
B-score difference between index and control knees
6 months
12 months
Index
Control
Difference
p
Difference
p
≥0 (All)
1.48 (78)
1.48 (78)
0.09
0.02
0.11
0.01
≥2
1.76 (62)
1.78 (57)
0.08
0.09
0.11
0.07
≥3
2.15 (48)
2.39 (41)
0.16
0.01
0.13
0.09
4
2.84 (27)
3.01 (26)
0.23
0.01
0.16
0.14 McGuire et al. Arthritis Research & Therapy (2021) 23:242 Page 7 of 9 Fig. 4 12-month femoral bone shape stabilization is associated with medial femoral condyle cartilage thickening/stabilization in the index knee. Subjects are stratified by B-score change quartiles. All error bars are standard errors of the mean Fig. 4 12-month femoral bone shape stabilization is associated with medial femoral condyle cartilage thickening/stabilization in the index knee. Subjects are stratified by B-score change quartiles. All error bars are standard errors of the mean knees from baseline to 6 months was more striking than
that from six months to the end of the study, suggesting
that a second series of injections would be a reasonable
consideration in planning the next clinical trial. Discussion In this study, we measured femoral bone shape using the
B-score metric that describes changes along a vector tra-
cing a linear path from a normal, non-osteoarthritic
state to the pathological shape of the osteoarthritic knee. Pathological bone shape change is characterized by the
broadening and flattening of the femoral condyles and
tibial plateaus and the growth of osteophytes. The vector
is defined by KLG = 0 at its origin and KLG ≥2, where
a higher B-score is more radiographically pathologic,
and, as demonstrated in previous work, is associated
with progressive knee OA. Knees treated with IA TPX-
100 had a bone shape change at 6 and 12 months similar
to that of knees classified as KLG = 0. This was substan-
tially different from the trajectory of control knees,
which was similar to that of knees classified as KLG ≥2. There were no significant baseline group differences in
any demographic, clinical, or MRI measures. The change
in slope among treated knees compared with control These data represent, to our knowledge, the first re-
port of an investigational agent demonstrating a signifi-
cant treatment difference compared with placebo on
pathologic bone shape (B-score) change in the knee. A
previous study of the novel Cathepsin K Inhibitor MIV-
711 demonstrated a reduction of pathological bone sur-
face area change in subjects with knee OA at 6 months,
with no treatment differences in clinical outcomes [17]. While clinically meaningful, treatment-related differ-
ences in B-score changes have not been defined, the dif-
ference in the rate of B-score change between TPX-100-
treated knees and placebo-exposed knees was notable,
particularly in the most severely osteoarthritic knees,
suggesting a slowing of the OA-related bone pathology
associated with OA progression and predictive of joint Table 4 Pearson correlations between femur bone shape stabilization and tibiofemoral cartilage thickening/stabilization in TPX-100-
treated knee
Change period
Knee compatment
n
Correlation coefficient
p-value
Baseline to 6 months
Entire femoral condyle
78
−0.206
0.07
Medial femoral condyle
78
−0.265
0.02
Entire TF
78
−0.193
0.09
Medial TF
78
−0.212
0.06
Baseline to 12 months
Entire femoral condyle
78
−0.303
0.007
Medial femoral condyle
78
−0.329
0.003
Entire TF
78
−0.296
0.009
Medial TF
78
– 0.320
0.004 s between femur bone shape stabilization and tibiofemoral cartilage thickening/stabilization in TPX-100- McGuire et al. Authors’ contributions DM was responsible for the conception and design of the study. AB was
responsible for initial drafting the article. DM, YK, MM, DR, AB, MB, and NS
provided critical revision of the article for important intellectual content. All
authors were responsible for analysis and interpretation of the data and for
final approval of the article and take responsibility for the integrity of the
work. Supplementary Information pp
y
The online version contains supplementary material available at https://doi. org/10.1186/s13075-021-02622-8. Additional file 1. AC-100 Promotes Cartilage Defect Repair In Vivo and
Chondrocyte Differentiation and Function In Vitro Acknowledgements g
The authors would like to thank Felix Eckstein, MD; Susanne Maschek, PhD;
Wolfgang Wirth, PhD; and the team at Chondrometrics for producing the
cartilage and bone measures, and Ali Guermazi, MD, PhD, Boston Imaging,
for MOAKS analyses. The authors would also like to thank Charles S. Davis,
PhD, for his invaluable contribution to the statistical analysis. All MRI
measures and statistical analyses were performed with monetary
compensation. Vendors and consultants have no equity or other interest in
OrthoTrophix, Inc. Abbreviations AAM: Active appearance models; BMI: Body mass index;
CONSORT: Consolidated standards of reporting trials; DESS: Double echo
steady state; DMOAD: Disease-modifying osteoarthritis drug; FFE: Fast field
echo (Philips); FLASH: Fast low-angle shot (Siemens); IA: Intra-articular;
ICRS: International cartilage repair society; KLG: Kellgren-Lawrence grading;
KOOS: Knee injury and osteoarthritis outcome score; MEPE: Matrix
extracellular phosphoglycoprotein; MOAKS: MRI osteoarthritis knee score;
MRI: Magnetic resonance imaging; NRS: Numerical rating scale;
OA: Osteoarthritis; OAI: Osteoarthritis Initiative; PF: Patellofemoral;
PFOA: Patellofemoral osteoarthritis; PPT: Per protocol; SAS: Statistical analysis
software; SDD: Smallest detectable difference; SIBLING: Small integrin-binding
ligand N-linked glycoprotein; SPGR: Spoiled gradient-recalled (GE);
TF: Tibiofemoral; 3D: Three dimensional; WOMAC: Western Ontario and
McMaster universities osteoarthritis index q
p
There were limitations in this study. TPX-100-5 was a
retrospective analysis of the subset of Study TPX-100-1
subjects including those with tibiofemoral OA, mild to
severe, in addition to PFOA, whose MRI images were
qualified for bone shape analyses. However, the statis-
tical analysis plan for the study was finalized prior to
image qualification and analysis, and all analyses were
performed blind to time point, treatment assignment,
and all clinical information. As bone-shape analysis was
not part of the original (TPX-100-1) study plan, the MR
image sequence used was designed for manual cartilage
segmentation and therefore was not optimal for auto-
mated bone segmentation and shape analysis, for which
thinner, 0.75-mm slices would have been preferred. Nevertheless, all the image data that could be utilized
were included in the analysis without any specific subject
selection. The severity of OA in the subject population
was defined by using MRI-based cartilage defect (ICRS)
scoring after clinical inclusion/exclusion criteria were
met. ICRS grading was performed by central readers
blind to clinical information in order to establish a base-
line of structural severity and to exclude significant
meniscal pathology and synovitis, per study exclusion
criteria. Discussion Arthritis Research & Therapy (2021) 23:242 Page 8 of 9 Table 5 Pearson correlations between femur bone shape stabilization and tibiofemoral cartilage thickening/stabilization in control
knee
Change period
Knee compartment
n
Correlation coefficient
p-value
Baseline to 6 months
Entire femoral condyle
78
−0.068
0.55
Medial femoral condyle
78
−0.053
0.65
Entire TF
78
0.043
0.71
Medial TF
78
0.011
0.93
Baseline to 12 months
Entire femoral condyle
78
−0.174
0.13
Medial femoral condyle
78
−0.246
0.03
Entire TF
76
0.011
0.92
Medial TF
77
−0.085
0.46 Table 5 Pearson correlations between femur bone shape stabilization and tibiofemoral cartilage thickening/stabilization in control combination with previously reported robust improve-
ments in established clinical outcomes (as measured by
WOMAC and KOOS scores), support the potential of
TPX-100 as a candidate disease-modifying drug in knee
OA. failure. In addition, treatment-associated reductions in
B-score correlated significantly with reduced cartilage
thickness loss in medial and total tibiofemoral cartilage
in TPX-100-treated knees at 12 months. TPX-100 acts on cells committed to hard tissue
lineage, viz. bone, cartilage, or dentin [2–4, 18]. Since
TPX-100 has been demonstrated to have positive effects
on both cartilage and bone formation, activities observed
in this clinical trial may be due to both activities. Based
on preliminary in vitro data, the putative mechanism of
action of TPX-100 is via integrin binding via the RGD
sequence found in TPX-100 (unpublished data). References 17. Conaghan PG, Bowes MA, Kingsbury S, Brett A, Guillard G, Rizoska B, et. al. Disease-Modifying Effects of a Novel Cathepsin K Inhibitor in Osteoarthritis. Ann Intern Med 2020;172(2):86-95. doi: https://doi.org/10.7326/M19-0675. 1. Hopwood B, Tsykin A, Findlay DM, Fazzalari NL. Microarray gene expression
profiling of osteoarthritic bone suggests altered bone remodelling, WNT
and transforming growth factor-β/bone morphogenic protein signalling. Arthritis Res Ther. 2007;9(5):R100. https://doi.org/10.1186/ar2301. 1. Hopwood B, Tsykin A, Findlay DM, Fazzalari NL. Microarray gene expression
profiling of osteoarthritic bone suggests altered bone remodelling, WNT
and transforming growth factor-β/bone morphogenic protein signalling. Arthritis Res Ther. 2007;9(5):R100. https://doi.org/10.1186/ar2301. 1. Hopwood B, Tsykin A, Findlay DM, Fazzalari NL. Microarray gene expression
profiling of osteoarthritic bone suggests altered bone remodelling, WNT
and transforming growth factor-β/bone morphogenic protein signalling. Arthritis Res Ther. 2007;9(5):R100. https://doi.org/10.1186/ar2301. 18. Liu H, Li W, Gao C, Kumagai Y, Blacher RW, DenBesten PK. Dentonin, a
Fragment of MEPE, Enhanced Dental Pulp Stem Cell Proliferation. J Dent
Res. 2004;83(6):496–9. https://doi.org/10.1177/154405910408300612. 18. Liu H, Li W, Gao C, Kumagai Y, Blacher RW, DenBesten PK. Dentonin, a
Fragment of MEPE, Enhanced Dental Pulp Stem Cell Proliferation. J Dent
Res. 2004;83(6):496–9. https://doi.org/10.1177/154405910408300612. Arthritis Res Ther. 2007;9(5):R100. https://doi.org/10.1186/ar2301. 2. Hayashibara T, Hiraga T, Yi B, Motoyoshi N, Kumagai Y, Nishimura R, et al. A
synthetic peptide fragment of human MEPE stimulates new bone formation
in vitro and in vivo. J Bone Miner Res. 2004;19(3):455–62. https://doi.org/1
0.1359/JBMR.0301263. Funding Initiative. Rheumatology 2016; 55:1585–1593. doi: https://doi.org/10.1093/
rheumatology/kew191. g
This study is sponsored by Orthotrophix, Inc., CA, USA. The study sponsor
was involved in the study design, collection, analysis, and interpretation of
data and in the writing of the manuscript and decision to submit for
publication. 9. Bowes MA, Vincent GR, Wolstenholme CB, Conaghan PG. A novel method
for bone area measurement provides new insights into osteoarthritis and its
progression. Ann Rheum Dis. 2013;74:519–25. https://doi.org/10.1136/a
nnrheumdis-2013-204052. 10. Hunter D, Nevitt M, Lynch J, Kraus VB, Katz JN, Collins JE, et. al. Longitudinal
validation of periarticular bone area and 3D shape as biomarkers for knee
OA progression? Data from the FNIH OA Biomarkers Consortium. Ann
Rheum Dis 2016; 75:1607–1614. doi: https://doi.org/10.1136/annrheumdis-2
015-207602. Availability of data and materials Osteoarthritis Initiative (OAI) source data are available from https://data-
archive.nimh.nih.gov/oai/. The datasets generated and/or analyzed within
this publication are available from the corresponding author on reasonable
request. 11. Bowes MA, Kacena K, Alabas OA, Brett A, Dube B, Bodick N, et.al. Machine-
learning, MRI bone shape and important clinical outcomes in osteoarthritis:
data from the Osteoarthritis Initiative. Ann Rheum Dis. 2020;80(4):502-508. doi: https://doi.org/10.1136/annrheumdis-2020-217160. Consent for publication Consent for publication
Not applicable. 14. Williams TG, Holmes A, Waterton J, Maciewicz R, Hutchinson CE, Moots R,
et al. Anatomically corresponded regional analysis of cartilage in
asymptomatic and osteoarthritic knees by statistical shape modelling of the
bone. IEEE Trans Med Imaging. 2010 Aug;29(8):1541–59. https://doi.org/1
0.1109/TMI.2010.2047653. Competing interests AB and MB are employees of Imorphics, Manchester, UK. DM, MM, DR, and
YK are employees of OrthoTrophix, Inc., Oakland, CA. NS reports no
competing interests. 15. Hunter DJ, Nevitt M, Lynch J, Kraus V, Katz JN, Collins J, et. al. Preliminary
assessment of predictive validity of periarticular bone area and shape
markers in knee OA. Osteoarthr Cartil 2013; 21: S175. doi: https://doi.org/1
0.1016/j.joca.2013.02.376. Author details
1 1OrthoTrophix, Foster City, CA 94404, USA. 2Imorphics, Manchester, UK. 3University of Kansas Medical Center, Kansas City, KS, USA. 16. Bowes MA, Lohmander LS, Wolstenholme CBH, Vincent GR, Conaghan PG,
Frobell RB. Marked and rapid change of bone shape in acutely ACL injured
knees – an exploratory analysis of the Kanon trial. Osteoarthr Cartil. 2019;
27(4):638–45. https://doi.org/10.1016/j.joca.2018.12.021. Ethics approval and consent to participate 12. Eckstein F, Ateshian G, Burgkart R, Burnstein D, Cicuttini F, Dardzinski B, et al. Proposal for a nomenclature for Magnetic Resonance Imaging based
measures of articular cartilage in osteoarthritis. Osteoarthr Cartil. 2006;14(10):
974–83. https://doi.org/10.1016/j.joca.2006.03.005. Written informed consent was obtained prior to study enrollment. Institutional Review Board approval was obtained from Western IRB with
approval ID #20131394. 13. Wirth W, Eckstein F. A technique for regional analysis of femorotibial
cartilage thickness based on quantitative magnetic resonance imaging. IEEE
Trans Med Imaging. 2008;27:737–44. https://doi.org/10.1109/TMI.2007. 907323. Conclusions These results of structural imaging in TPX-100-treated
knees
compared
with
placebo-exposed
knees,
in Page 9 of 9 McGuire et al. Arthritis Research & Therapy (2021) 23:242 Received: 19 February 2021 Accepted: 5 September 2021 Received: 19 February 2021 Accepted: 5 September 2021 Publisher’s Note 3. Nagel DE, Khosla S, Sanyal A, Rosen DM, Kumagai Y, Riggs L. A fragment of
hypophospatemic factor, MEPE, requires inducible cyclooxygenase-2 to
exert potent anabolic effects on normal human marrow osteoblast
precursors. J Cell Biochem. 2004;93(6):1107–14. https://doi.org/10.1002/jcb.2
0249. Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations. 4. Middleton-Hardie CA, Aberman H, Simon T, Alliston T, Mortazavi A, Rosen
DM. AC-100 Promotes Cartilage Defect Repair In Vivo and Chondrocyte
Differentiation and Function In Vitro [abstract]. Proceedings of the 56th
Annual Meeting of the Orthopedic Research Society, poster # 0974. Presented March 6, 2010. New Orleans, LA 4. Middleton-Hardie CA, Aberman H, Simon T, Alliston T, Mortazavi A, Rosen
DM. AC-100 Promotes Cartilage Defect Repair In Vivo and Chondrocyte
Differentiation and Function In Vitro [abstract]. Proceedings of the 56th
Annual Meeting of the Orthopedic Research Society, poster # 0974. Presented March 6, 2010. New Orleans, LA 5. McGuire D, Lane N, Segal N, Metyas S, Barthel H, Miller M, et al. TPX-100
leads to marked, sustained improvements in subjects with knee
osteoarthritis: pre-clinical rationale and results of a controlled clinical trial
[abstract]. Osteoarthr Cartil. 2018;26:S243. https://doi.org/10.1016/j.joca.2018. 02.50219. 5. McGuire D, Lane N, Segal N, Metyas S, Barthel H, Miller M, et al. TPX-100
leads to marked, sustained improvements in subjects with knee
osteoarthritis: pre-clinical rationale and results of a controlled clinical trial
[abstract]. Osteoarthr Cartil. 2018;26:S243. https://doi.org/10.1016/j.joca.2018. 02.50219. 6. McGuire D, Segal N, Metyas S, Barthel H, Miller M, Rosen D, et al. Intra-
articular TPX-100 in knee osteoarthritis: robust functional response at 6 and
12 months is associated with increased tibiofemoral cartilage thickness
[abstract L16]. Arthritis Rheumatol. 2018;70(suppl 10):3405-3406. 7. Neogi T, Bowes MA, Niu J, De Souza KM, Vincent GR, Goggins J, et. al. Magnetic Resonance Imaging-Based Three-Dimensional Bone Shape of the
Knee Predicts Onset of Knee Osteoarthritis: Data From the Osteoarthritis
Initiative. Arthritis Rheum 2013;65(8):2048-2058. doi: https://doi.org/10.1
002/art.37987. 8. Barr AJ, Dube B, Hensor E, Kingsbury S, Peat G, Bowes M, et. al. The
relationship between three-dimensional knee MRI bone shape and total
knee replacement—a case control study: data from the Osteoarthritis
|
https://openalex.org/W3115118972
|
https://smw.ch/index.php/smw/article/download/2919/4793
|
English
| null |
Early experimental COVID-19 therapies: associations with length of hospital stay, mortality and related costs
|
Schweizerische medizinische Wochenschrift
| 2,020
|
cc-by
| 7,403
|
Correspondence:
Stéphanie Baggio, MD,
Geneva University Hospi-
tals, Division of Prison
Health, Chemin du Petit Bel
Air 2, CH-1226 Thônex,
stephanie.bag-
gio[at]hcuge.ch Original article | Published 31 December 2020 | doi:10.4414/smw.2020.20446
Cite this as: Swiss Med Wkly. 2020;150:w20446 Summary 1.37–6.12, p = 0.002), 1.23 additional hospitalisation days
(95% CI −1.24 – 3.51, p = 0.319), and 4.19 additional
hospitalisation days (95% CI 1.52–5.31, p <0.001), re-
spectively, compared with patients treated with the stan-
dard of care. Neither experimental therapy was significant-
ly associated with mortality. These additional hospital days
amounted to 1010.77 additional days for hydroxychloro-
quine and hydroxychloroquine combined with lopinavir/ri-
tonavir, resulting in an additional cost of US$ 2,492,214
(95%CI US$ 916,839–3,450,619). 1.37–6.12, p = 0.002), 1.23 additional hospitalisation days
(95% CI −1.24 – 3.51, p = 0.319), and 4.19 additional
hospitalisation days (95% CI 1.52–5.31, p <0.001), re-
spectively, compared with patients treated with the stan-
dard of care. Neither experimental therapy was significant-
ly associated with mortality. These additional hospital days
amounted to 1010.77 additional days for hydroxychloro-
quine and hydroxychloroquine combined with lopinavir/ri-
tonavir, resulting in an additional cost of US$ 2,492,214
(95%CI US$ 916,839–3,450,619). AIMS OF THE STUDY: Hydroxychloroquine and lopinavir/
ritonavir have been used as experimental therapies to
treat COVID-19 during the first wave of the pandemic. Randomised controlled trials have recently shown that
there are no meaningful benefits of these two therapies
in hospitalised patients. Uncertainty remains regarding the
potential harmful impact of these therapies as very early
treatments and their burden to the health care system. The present study investigated the length of hospital stay
(LOS),
mortality,
and
costs
of
hydroxychloroquine,
lopinavir/ritonavir or their combination in comparison with
standard of care among patients hospitalised for coron-
avirus disease 2019 (COVID-19). CONCLUSIONS: Prescribing experimental therapies for
COVID-19 was not associated with a reduced LOS and
might have increased the pressure put on healthcare sys-
tems. Keywords: COVID-19, healthcare costs, hydroxychloro-
quine,
lopinavir/ritonavir,
observational
study,
SARS-
CoV-2 METHODS: This retrospective observational cohort study
took place in the Geneva University Hospitals, Geneva,
Switzerland (n = 840) between 26 February and 31 May
2020. Demographics, treatment regimens, comorbidities,
the modified National Early Warning Score (mNEWS) on
admission, and contraindications to COVID-19 treatment
options were assessed. Outcomes included LOS, in-hos-
pital mortality, and drug and LOS costs. Early experimental COVID-19 therapies:
associations with length of hospital stay,
mortality and related costs Vernaz Nathaliea, Agoritsas Thomasbc, Calmy Alexandrad, Gayet-Ageron Angèlee, Gold Gabrielf, Perrier Arnaudgh, Picard
Fabiennei, Prendki Virginiej, Reny Jean-Luch, Samer Carolinek, Stirnemann Jérômeh, Vetter Paulinel, Zanella Marie-Célinem,
Zekry Dinaj, Baggio Stéphanieno a Medical Directorate, Finance Directorate, Geneva University Hospitals, Geneva University, Switzerland
b Division of General Internal Medicine and Division of Clinical Epidemiology, University Hospitals of Geneva, Switzerland Medical Directorate, Finance Directorate, Geneva University Hospitals, Geneva University, Switzerland
Di i i
f G
l I t
l M di i
d Di i i
f Cli i
l E id
i l
U i
it H
it l
f G
S it
l
d Medical Directorate, Finance Directorate, Geneva University Hospitals, Geneva University, Switzerland
b Division of General Internal Medicine and Division of Clinical Epidemiology, University Hospitals of Geneva, Switzerland
c Department of Health Research Methods, Evidence, and Impact, McMaster University, Hamilton, ON, Canada p
,
,
p
,
y,
d Division of Infectious Diseases, HIV/AIDS Unit, Geneva University Hospitals, Switzerland y
CRC and Division of Clinical Epidemiology, Department of Health and Community Medicine, University of Geneva and Un
Switzerland f
Service of Geriatrics, Department of Internal Medicine Rehabilitation and Geriatrics, University Hospitals and University of Geneva, Switzerland Service of Geriatrics, Department of Internal Medicine Rehabilitation and Geriatrics, University Hospitals and University of Gen epartment of Clinical Neurosciences, University of Geneva and University Hospitals of Geneva, Switzerland
cine for the Aged, University of Geneva and University Hospitals of Geneva, Thônex, Switzerland Division of Clinical Pharmacology and Toxicology, Geneva University Hospitals, Switzerland
Geneva Centre for Emerging Viral Diseases, Geneva University Hospitals, Switzerland n Division of Prison Health, University of Geneva and University Hospitals of Geneva, Thônex, Switzerland
o Office of Corrections, Department of Justice and Home Affairs of the Canton of Zurich, Zurich, Switzerland n Division of Prison Health, University of Geneva and University Hospitals of Geneva, Thônex, Switzerland
o Office of Corrections Department of Justice and Home Affairs of the Canton of Zurich Zurich Switzerland n Division of Prison Health, University of Geneva and University Hospitals of Geneva, Thônex, Switzerland
o Office of Corrections, Department of Justice and Home Affairs of the Canton of Zurich, Zurich, Switzerland Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”.
No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Measurements Clinical outcomes: The LOS for patients who were read-
mitted for reasons related to COVID-19 within 18 days of
the first discharge was aggregated [13]. We also collected
in-hospital mortality. Study design and setting This study was a single-centre retrospective cohort study
performed during the first wave of the pandemic (February
to May 2020 [8]), between 26 February and 31 May 2020,
with patients diagnosed with COVID-19 who were ad-
mitted to acute medical wards in internal medicine and
geriatrics, including intermediate care and intensive care
at Geneva University Hospitals (HUG), Geneva, Switzer-
land. All patients were hospitalised in COVID-19 wards. HUG is the leading Swiss university hospital and the only
public hospital in the canton of Geneva, with 64,000 hospi-
talisations and 1.1 million outpatient visits per year. Ethics
approval was granted by the Cantonal Ethics Research
Committee of Geneva (REF 2020-01070). Additional COVID-19-related costs: The additional costs
of the experimental therapies, including direct drug costs
and costs associated with an extended LOS. In the absence
of specific COVID-19 hospitalisation costs, we chose the
most recent year of reference (2018). Additional hospital-
isation costs were measured according to the 2018 Swiss
accounting REKOLE® system. The 2018 mean cost per pa-
tient per day hospitalised was US$ 1152 (EUR 983, CHF
1061) for geriatric wards and US$ 2568 (EUR 2190, CHF
2365) for internal medicine wards. The cost calculation
considered the direct costs of the different lopinavir/riton-
avir regimens adapted for patients over 75 years of age and
the direct hospitalisation costs. Public prices for the drugs
were extracted from the official Swiss prices. Costs were
converted from Swiss francs to dollars and euros, at the ex-
change rate of 1 CHF = US$ 1.09 = EUR 0.93 (1 October
2020). Introduction During the first wave of the severe acute respiratory syn-
drome coronavirus 2 (SARS-CoV-2) pandemic, the ma-
jority of countries experienced severe healthcare, social,
and economic challenges. Huge efforts were directed to-
wards finding an effective treatment to reduce morbidity
and mortality. This study was conducted in Switzerland,
a country severely affected by the pandemic, with one of RESULTS: After successful propensity score matching,
patients treated with (1) hydroxychloroquine, (2) lopinavir/
ritonavir or (3) their combination had on average 3.75 ad-
ditional hospitalisation days (95% confidence interval [CI] Page 1 of 9 Swiss Medical Weekly · PDF of the online version · www.smw.ch Swiss Medical Weekly · PDF of the online version · www.smw.ch Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”. No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Original article Swiss Med Wkly. 2020;150:w20446 31 May 31 2020; (3) hospitalisation for reasons other than
COVID-19, such as delivery, emergency surgery, presence
on a waiting list for nursing home and psychiatric in-
patient, hospitalisation for isolation; and (4) the use of
other COVID-19 experimental therapies (e.g., remdesivir,
anakinra, azithromycin). the highest incidence rates of SARS-CoV-2 infections per
capita [1], a total of 30,762 infected individuals and 1,656
deaths during the first wave (February to May 2020). We
assessed two repositioned drugs that were commonly used
worldwide in first wave of the pandemic: hydroxychloro-
quine, lopinavir/ritonavir, and their combination. Overall, randomised controlled trials have thus far demon-
strated no meaningful benefits of these two therapies in
patients hospitalised for severe or critical COVID-19, as
summarised in a recent living network meta-analysis
[2–4]. The hydroxychloroquine and lopinavir/ritonavir
arms of a major randomised controlled trial were stopped
in June 2020 due to the absence of clinical benefit over the
standard of care [3–7]. However, real-life data are crucial-
ly needed to gain a better understanding of the potential
harmful impact of these drugs and their burden on the
healthcare system. Our study thus aimed to compare hy-
droxychloroquine, lopinavir/ritonavir, and their combina-
tion when routinely used during the first wave of Covid-19
with standard of care without specific antiviral therapy in
terms of (1) length of hospital stay (LOS), (2) in-hospital
mortality, and (3) related additional healthcare costs. Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”.
No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Experimental therapies for Covid-19 The experimental therapies, given alone or in combination,
were used at the following dosages, based on institutional
recommendations [11, 12]: (1) 400 mg of lopinavir/riton-
avir twice daily for 5 days under the age of 75 years and
400 mg of lopinavir/ritonavir in the morning and 200 mg
of lopinavir/ritonavir in the evening for patients over 75;
(2) a single dose of 800 mg of hydroxychloroquine; and (3)
lopinavir/ritonavir and hydroxychloroquine dual therapy at
the same dosages. Lopinavir/ritonavir was first prescribed
on 5 March 2020, and hydroxychloroquine alone or com-
bined with lopinavir/ritonavir on 20 March 2020. Institu-
tional recommendations did not change over the study pe-
riod. Swiss Medical Weekly · PDF of the online version · www.smw.ch Participants We included all patients from 26 February to 30 April
2020, who were discharged from acute medical wards by
31 May 2020. Patients were included if they were hospi-
talised for a SARS-CoV-2 infection, with either a positive
reverse-transcriptase polymerase chain reaction test (RT-
PCR) for SARS-CoV-2 or a negative RT-PCR but a high
suspicion of COVID-19 based on a plausible clinical pre-
sentation, chest imaging or the presence of SARS-CoV-2
IgG as a marker of past SARS-CoV-2 infection [9]. Hos-
pitalisation criteria were standardised as much as possible
in routine care and compatible with the World Health Or-
ganization (WHO) definition of either severe or critical
COVID-19: pneumonia with CURB-65 score ≥2, sustained
respiratory rate >20/min, need for oxygen (or increased re-
quirement for oxygen) or respiratory distress, alternation
of general status or rapid worsening, or other acute co-ex-
isting condition. These criteria remained the same from the
beginning of the first wave and over the study period. Administrative data included age, gender, dates of admis-
sion and discharge, acute care in internal medicine and
geriatrics, and the month of hospitalisation (February and
March aggregated or April). COVID-19 severity: The modified National Early Warning
Score (mNEWS) adapted for use in COVID-19 was used
to assess COVID-19 severity [14]. Age (<65 or ≥65years),
mean respiratory rate, heart rate, systolic blood pressure
and oxygen saturation, administration of supplemental
oxygen, level of consciousness, and the highest tempera-
ture during the first 24 hours were used to compute the
mNEWS, which ranged from 0 to 23. Comorbidities: We extracted the relevant medical history,
including the presence of active asthma, chronic obstruc-
tive pulmonary disease or sleep apnoea, cancer (previous
5 years), renal impairment, diabetes, hypertension, heart
disease, and cirrhosis. Comorbidities were summed and
the total score ranged from 0 to 8 comorbidities. Obesity The following exclusion criteria were used: (1) death, intu-
bation, transfer from other hospitals or discharge within the
first 24 hours of hospitalisation [10]; (2) not discharged on Swiss Medical Weekly · PDF of the online version · www.smw.ch Page 2 of 9 Page 2 of 9 Original article Swiss Med Wkly. 2020;150:w20446 ties). Statistical significance was assessed at a two-sided
0.05 level for all analyses. (body mass index [BMI] ≥30 kg/m2) was also recorded and
analysed separately from other comorbidities. Participants Contraindications: Medications that should not be co-ad-
ministered with hydroxychloroquine and/or lopinavir/ri-
tonavir were extracted on the basis of the Liverpool recom-
mendations. QT interval corrected with the Bazett formula
before the administration of any therapy or at admission
when a therapy had already been administered was also
extracted. Two binary variables were created: (1) hydrox-
ychloroquine contraindications (medications and/or cor-
rected QT ≥500 ms), and (2) lopinavir/ritonavir contraindi-
cations (medications). Hydroxychloroquine The top panel of table 2 reports the associations of factors
before and after propensity score matching. In the entire
cohort (hydroxychloroquine/standard of care, n = 599), the
probability of receiving hydroxychloroquine was signifi-
cantly higher in males (p = 0.007), patients hospitalised in
internal medicine (p <0.001), and patients with a higher
mNEWS (p <0.001, left panel of table 2). After propensity
score matching, there were no significant differences be-
tween groups (right panel of table 2). We performed deterministic sensitivity analyses to exam-
ine the robustness of our findings. We tested whether the
results were similar using other propensity score methods:
(1) covariate adjustment using the propensity score, (2)
stratification based on the propensity score, (3) inverse
probability weighting using the propensity score, and (4)
nearest-neighbour matching using random draw. We also
used the log of the LOS instead of the original count out-
come. Finally, analyses were performed for different sub-
samples: (1) after the exclusion of cases not confirmed
with RT-PCR, (2) after the exclusion of the most severe
cases (patients admitted to the intensive care unit, patients
who died), and (3) after exclusion of participants who had
contraindications to the corresponding experimental ther-
apy. Results were similar to those reported in the Results
section. After propensity score matching, hydroxychloroquine
treatment was significantly associated with a longer LOS,
with on average 3.75 additional days compared with
matched patients treated with the standard of care (95% CI
1.37–6.12, p = 0.002). There was no significant difference
in mortality between the two groups, with a difference of
–0.02% on average (95% CI –0.14 – 0.09, p = 0.712). Characteristics of the cohort The study flowchart is depicted in figure 1. Among the
1113 patients who were admitted to HUG wards from 26
February to 30 April 2020, 1059 had a positive RT-PCR
for SARS-CoV-2 and 54 patients tested negative but were
strongly suspected of having the disease based on medical
judgement. A total of 273 patients were excluded from the
study because they were ineligible (see detail in fig. 1). Out
of the 840 patients included in our analysis, 93 received
hydroxychloroquine, 83 received lopinavir/ritonavir, 158
received hydroxychloroquine and lopinavir/ritonavir, and
506 received neither drug. Statistical analysis We first described the cohort using descriptive statistics
and tested which factors were associated with the out-
comes using simple negative binomial (LOS) and logistic
(in-hospital mortality) regressions. We then investigated
the associations of experimental COVID-19 therapies with
LOS and in-hospital mortality. We used propensity score
matching analyses to minimise the effects of confounding
factors. Propensity score matching was performed sep-
arately for each therapy (1) hydroxychloroquine, (2)
lopinavir/ritonavir, and (3) hydroxychloroquine combined
with lopinavir/ritonavir) compared with the standard of
care. For each treatment group, the propensity score was
estimated using a multiple probit regression that included
the following factors: gender, location of hospitalisation,
month of hospitalisation, mNEWS, number of comorbidi-
ties, BMI, and presence of contraindications for hydroxy-
chloroquine (for analyses on hydroxychloroquine and hy-
droxychloroquine combined with lopinavir/ritonavir) or
lopinavir/ritonavir (for analyses of lopinavir/ritonavir and
hydroxychloroquine combined with lopinavir/ritonavir). We created separate matched control groups for each ther-
apy using nearest-neighbour matching (full Mahalanobis). Multiple neighbours were allowed when controls had iden-
tical propensity scores. We calculated an average treatment
effect on the treated for each outcome. Confidence inter-
vals (CIs) were computed using bootstrapping. Crude as-
sociations (without propensity score matching) and asso-
ciations for the matched groups (after propensity score
matching) are reported for each therapy. Descriptive statistics for the entire cohort are detailed in
table 1. The mean age of included patients was 67.90
years (standard deviation [SD] 18.77), 50.5% were male
(n = 424), and the mNEWS was 6.46 (SD 2.88) at admis-
sion. Most patients were hospitalised in internal medicine
(69.9%, n = 587) and during the month of February/March
2020 (66.0%, n = 554). The average LOS was 10.38 days
(SD 7.98), and a total of 15.6% (n = 131) patients died. Patients were more likely to receive lopinavir/ritonavir in
February and March. The associations of variables includ-
ed in the propensity scores with the outcomes are shown in
supplementary table S1 (appendix 1). Before the propensi-
ty score matching, factors that were significantly associat-
ed with longer LOS included male gender, older age, hos-
pitalisation in geriatrics, hospitalisation in April, a higher
mNEWS, a high number of comorbidities, and having con-
traindications for hydroxychloroquine. Factors associated
with mortality included male gender, older age, hospitali-
sation during the month of March, a higher mNEWS, and
a high number of comorbidities. Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”.
No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Swiss Medical Weekly · PDF of the online version · www.smw.ch Lopinavir/ritonavir The second panel of table 2 reports the associations of
factors before and after propensity matching. In the entire
cohort (lopinavir/ritonavir/standard of care, n = 589), the
probability of receiving lopinavir/ritonavir was significant- Analyses were performed using Stata 15 (pscore for
propensity score estimation, no imposition of common
support and psmatch2 for propensity matching, option Swiss Medical Weekly · PDF of the online version · www.smw.ch Page 3 of 9 Page 3 of 9 Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”. No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Original article Swiss Med Wkly. 2020;150:w20446 significantly higher in males (p <0.001), patients in inter-
nal medicine (p <0.001), patients with higher mNEWS (p
<0.001), patients with more comorbidities (p <0.001), and
patients without contraindications for both hydroxychloro-
quine (p <0.001) and lopinavir/ritonavir (p <0.001, left
panel of table 2). After propensity score matching, no sig-
nificant differences between groups remained (right panel
of table 2). ly higher in males (p = 0.050), patients hospitalised in in-
ternal medicine (p <0.001), patients hospitalised in March
(p <0.001), patients with more comorbidities (p = 0.016),
and patients with no contraindications for lopinavir/riton-
avir (p < 0.001, left panel of table 2). After propensity
score matching, no significant differences between groups
remained (right panel of table 2). After propensity score matching, the LOS was not signif-
icantly different between patients who received lopinavir/
ritonavir and patients who received standard of care (1.23
days, 95% CI −1.24 – 3.51, p = 0.319). Mortality was also
not significantly different, with an average difference of
−0.05% (95% CI −0.19 – 0.02, p = 0.639). After propensity score matching, hydroxychloroquine
combined with lopinavir/ritonavir was significantly asso-
ciated with an average of 4.19 days longer LOS compared
with patients treated with the standard of care (95% CI
1.52–5.31, p <0.001). There was no significant difference
in mortality between the two groups, with a difference of
−0.05% on average (95% CI −0.15–0.05, p =0.697). Main findings Our study showed no significant beneficial association be-
tween LOS and the administration of either hydroxy-
chloroquine, lopinavir/ritonavir or their use in combination
among
hospitalised
patients
with
severe
or
critical
COVID-19, in line with recently published findings of ran-
domised controlled trials and observational studies [3, 4,
10, 15, 16]. In particular, none of the regimens was associ-
ated with shortened hospital stays or higher survival, com- Table 1: Baseline characteristics and outcomes of the full study cohort. Additional drugs and LOS costs The third panel of table 2 reports the associations of factors
before and after propensity matching. In the entire cohort
(hydroxychloroquine combined with lopinavir/ritonavir/
standard of care, n = 664), the probability of receiving hy-
droxychloroquine combined with lopinavir/ritonavir was Additional drugs and LOS costs Table 3 reports results of the cost analysis. The total ad-
ditional days of hospitalisation were 348.75 (95% CI
127.41–569.16) for hydroxychloroquine and 662.02 addi-
tional days (95% CI 240.16–838.98) for hydroxychloro- Figure 1: Flow chart of the study cohort. RT-PCR = reverse-transcriptase-polymerase-chain reaction; HCQ = hydroxychloroquine; LPV/r =
lopinavir–ritonavir; AZM = azithromycin. of the online version · www.smw.ch
ht license “Attribution – Non-Commercial – No Derivatives 4 0”
Page 4 of 9 Swiss Medical Weekly · PDF of the online version · www.smw.ch Swiss Medical Weekly · PDF of the online version · www.smw.ch Page 4 of 9 Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”. No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”. No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Original article Swiss Med Wkly. 2020;150:w20446 pared with COVID-19 patients treated with the standard of
care. On the contrary, hydroxychloroquine (alone or when
combined with lopinavir/ritonavir) was significantly asso-
ciated with an increased LOS. Treatment with lopinavir/ri-
tonavir alone did not demonstrate any statistically signifi-
cant association with LOS. quine combined with lopinavir/ritonavir, accounting for a
total of 1010.77 additional days (95% CI 367.57–1408.14)
for hydroxychloroquine and hydroxychloroquine com-
bined with lopinavir/ritonavir. Costs associated with
lopinavir/ritonavir were not included, as there was no sig-
nificant association between LOS and prescription of
lopinavir/ritonavir. Using the different costs for one day of
hospitalisation in internal medicine or geriatric wards, the
total additional costs were US$ 816,073 (EUR 695,984,
CHF 751,637) (95% CI US$ 298,211–1,331,760; EUR
254,328–1,135,780; CHF 274,665–1,226,610) for hydrox-
ychloroquine and US$ 1,676,141 (EUR 1,429,490, CHF
1,543,790)
(95%
CI
US$
618,629–2,118,859;
EUR
527,595–1,807,060; CHF 569,783–1,951,560) for hydrox-
ychloroquine plus lopinavir/ritonavir, accounting for a to-
tal
cost
of
US$
2,492,214
(EUR
2,125,470,
CHF
2,295,430)
(95%
CI
US$
916,839–3,450,619;
EUR
781,921–2,942,840; CHF 844,446–3,178,160). Regarding the findings on hydroxychloroquine (alone or in
combination with lopinavir/ritonavir), a first hypothesis is
that these regimens would worsen COVID-19 or resulted
in complications. Additional drugs and LOS costs Hydroxychloroquine is an old and af-
fordable medication that was widely promoted and en-
dorsed worldwide, based on very low level of scientific
evidence, at the time of the study: in-vitro studies and stud-
ies with important methodological limitations, such as lack
of control group and small number of patients included
or bias [17–20]. Even though our study used a hydroxy-
chloroquine single dose of 800 mg, lower than in other
studies (e.g., RECOVERY: 2.4 g during the first 24 hours
and a cumulative dose of 9.2 g over 10 days; SOLIDARI-
TY: cumulative dose of 4 g over 10 days [21]), we found a
significant increase of LOS. Discussion A second hypothesis is that patients would be more likely
to be discharged later in order to monitor potential adverse
events, in an unprecedented political and media context
that placed physicians under extraordinary pressure to pre-
scribe these therapies off-label [22]. Of note, the literature
highlighted the hydroxychloroquine safety risks that could
lead to QT interval prolongation or adverse events [17,
20, 23–25]. Despite local guidance requiring that hydrox-
ychloroquine could only be prescribed in absence of con-
traindications, about 22% of patients who had contraindi- Swiss Medical Weekly · PDF of the online version · www.smw.ch Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”. cations
nevertheless
received
it.
This
difference
is
important because the toxicity may outweigh potential Feb./March)*
0.31
0.37
0.182
00.31
00.30
0.807
− mNEWS†
7.39
6.06
<0.001
70.39
70.25
0.621
− Number of diseases†
1.13
1.80
<0.001
10.13
10.04
0.539
− BMI ≥ 30*
0.18
0.17
0.752
00.18
00.16
0.653
− Contraindications HCQ*
0.07
0.24
<0.001
00.07
00.08
0.829
− Contraindications LPV/r*
0.08
0.34
<0.001
00.08
00.03
0.081
Outcomes
− LOS
11.57
10.13
0.044
110.57
70.38
<0.001
− In-hospital mortality
0.10
0.19
0.013
00.10
00.12
0.697
BMI = body mass index; HCQ = hydroxychloroquine; LOS = length of stay); LPV/r = lopinivir/ritonavir; mNEWS = modified National Early Warning Score; SOC = standard of care Propensity-based matched cohort BMI = body mass index; HCQ = hydroxychloroquine; LOS = length of stay); LPV/r = lopinivir/ritonavir; mNEWS = modified National Early Warning Score; SOC = standard of care
* Proportions are reported. † Means are reported. Table 3: Total additional cost of LOS for HCQ and HCQ combined with LPV/r in US$. HCQ
HCQ + LPV/r
Extra LOS (95% CI)
3.75 (1.37–6.12)
4.19 (1.52–5.31)
Internal medicine
Number of patients
78
151
Additional LOS (95% CI)
292.5 (106.86–477.36)
632.69 (229.52–801.81)
Additional cost of LOS (95% CI) in $
751,152 (274,421–1,225,880)
1,624,773 (589,417–2,059,080)
Cost of experimental therapies in $
96
16,954
Geriatrics
Number of patients
15
7
Additional LOS (95% CI)
56.25 (20.55–91.8)
29 (10.64:3 7.17)
Additional cost of LOS (95% CI) in $
64,807 (23,676–105,766)
33,792 (12,059–42,825)
Cost of experimental therapies in $
18
621
Total additional LOS (95% CI)
348.75 (127.41–569.16)
662.02 (240.16–838,98)
Total additional cost in $
816,073 (298,211–1,331,760)
1,676,141 (618,629–2,118,859)
CI = confidence interval; HCQ = hydroxychloroquine; LOS = length of stay; LPV/r = lopinavir/ritonavir
Swiss Medical Weekly · PDF of the online version · www.smw.ch
Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”. No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Page 6 of 9 Table 3: Total additional cost of LOS for HCQ and HCQ combined with LPV/r in US$. Main findings Whole cohort
SOC
HCQ
LPV/r
HCQ + LPV/r
n = 840
n = 506
n = 93
n = 83
n = 158
Gender†
− Male
50.5 (424)
43.9 (222)
59.1 (55)
55.4 (46)
36.1 (57)
− Female
49.5 (416)
56.1 (284)
40.9 (38)
44.6 (37)
63.9 (101)
Age*
67.90 (18.77, 0–101)
70.75 (20.01, 0–101)
66.14 (15.77, 16–96)
63.40 (17.40, 28–90)
62.15 (14.77, 21–95)
Location†
− Main hospital
69.9 (587)
57.7 (292)
83.9 (78)
79.5 (66)
95.6 (151)
− Hospital for elderly people
30.1 (253)
42.3 (214)
16.1 (15)
20.5 (17)
4.4 (7)
Month of hospitalisation†
− February/March
66.0 (554)
63.2 (320)
54.8 (51)
89.2 (74)
69.0 (109)
− April
34.0 (286)
36.8 (186)
45.2 (42)
10.8 (9)
31.0 (49)
mNEWS (0–23)*
6.46 (2.88, 0–15)
6.01 (2.96, 0–15)
7.21 (2.86, 3–13)
6.58 (2.59, 1–13)
7.39 (2.49, 1–14)
Number of diseases (0–8)*
1.59 (1.46, 0–7)
1.75 (1.52, 0–5)
1.72 (1.39, 0–5)
1.31 (1.46, 0–5)
1.13 (1.20, 0–5)
− Pulmonary diseases†
13.2 (111)
13.4 (68)
16.1 (15)
12.1 (10)
11.4 (18)
− Asthma†
3.6 (30)
4.6 (23)
2.2 (2)
1.2 (1)
2.5 (4)
− Cancer†
9.6 (81)
11.7 (59)
8.6 (8)
7.2 (6)
5.1 (8)
− Renal impairment†
26.7 (224)
30.2 (153)
25.8 (24)
22.9 (19)
17.7 (28)
− Diabetes†
20.6 (173)
20.8 (105)
22.6 (21)
20.5 (17)
19.0 (30)
− Hypertension†
48.5 (407)
50.0 (253)
55.9 (52)
42.2 (35)
42.4 (67)
− Heart diseases†
31.1 (261)
37.8 (191)
34.4 (32)
19.3 (16)
13.9 (22)
− Liver diseases†
5.2 (44)
6.3 (32)
6.5 (6)
6.0 (5)
0.6 (1)
BMI ≥30 kg/m2†
17.7 (149)
16.6 (84)
22.6 (21)
19.3 (16)
17.7 (28)
Contraindications†
− HCQ
19.1 (153)
24.1 (113)
21.5 (20)
11.1 (9)
7.0 (11)
− LPV/r
26.2 (220)
33.0 (167)
35.5 (33)
9.6 (8)
7.6 (12)
LOS*
10.38 (7.98, 1.02–56.71)
9.93 (7.78, 1.02–51.75)
12.48 (9.56, 1.92–56.71)
8.55 (6.98, 1.25–33.92)
11.57 (8.07, 1.89–53.52)
Mortality†
15.6 (131)
18.6 (94)
12.9 (12)
10.8 (9)
10.1 (16)
BMI = body mass index; HCQ = hydroxychloroquine; LOS = length of stay); LPV/r = lopinivir/ritonavir; mNEWS = modified National Early Warning Score; SOC = standard of care
* Means and (standard deviations with minimum and maximum) are reported. † Percentages and (n) are reported. Swiss Medical Weekly · PDF of the online version · www.smw.ch Page 5 of 9 Page 5 of 9 Original article Swiss Med Wkly. 2020;150:w20446 cations
nevertheless
received
it.
This
difference
is
important because the toxicity may outweigh potential Table 2: Associations of factors and outcomes with HCQ, LPV/r, and their combination treatment before and after propensity score matching. Overall cohort
Propensity-based matched cohort
HCQ
SOC
p-value
HCQ
SOC
p-value
n = 93
n = 506
n = 93
n = 105
Variables of the propensity score
− Gender (ref. female)*
0.59
0.44
0.007
00.59
00.60
0.882
− Location (ref. internal medicine)*
0.16
0.43
<0.001
00.16
00.13
0.535
− Month of hospitalisation (ref. Feb./March)*
0.45
0.37
0.134
00.45
00.50
0.559
− mNEWS†
7.22
6.06
0.001
70.22
60.76
0.302
− Number of diseases†
1.72
1.80
0.627
10.72
10.62
0.661
− BMI ≥30 kg/m2*
0.23
0.17
0.169
00.23
00.23
10.00
− Contraindications HCQ*
0.22
0.24
0.592
00.22
00.16
0.351
Outcomes
− LOS
12.48
10.13
0.010
120.48
80.73
0.002
− In-hospital mortality
0.13
0.19
0.191
00.13
00.15
0.712
LPV/r
SOC
p-value
LPV/r
SOC
p-value
n = 83
n = 506
n = 83
n = 98
Variables of the propensity score
− Gender (ref. female)*
0.55
0.44
0.050
00.55
00.49
0.440
− Location (ref. internal medicine)*
0.21
0.42
<0.001
00.21
00.15
0.310
− Month of hospitalisation (ref. Feb./March)*
0.11
0.37
<0.001
00.11
00.08
0.602
− mNEWS†
6.58
6.01
0.099
60.58
60.34
0.537
− Number of diseases†
1.31
1.75
0.016
10.31
10.28
0.873
− BMI ≥30 kg/m2*
0.19
0.17
0.548
00.19
00.12
0.202
− Contraindications LPV/r*
0.10
0.33
<0.001
00.10
00.10
10.00
Outcomes
− LOS
8.55
9.93
0.124
80.55
70.32
0.319
− In-hospital mortality
0.11
0.19
0.086
00.11
00.08
0.639
HCQ + LPV/r
SOC
p-value
HCQ + LPV/r
SOC
p-value
n = 158
n = 506
n = 158
n = 117
Variables of the propensity score
− Gender (ref. female)*
0.64
0.44
<0.001
00.64
00.58
0.251
− Location (ref. internal medicine)*
0.04
0.43
<0.001
00.04
00.04
0.778
− Month of hospitalisation (ref. Limitations of this study This study had important limitations that should be con-
sidered in the interpretation of these findings. First, we
excluded patients with severe COVID-19 who were intu-
bated at admission and most of the patients in the inten-
sive care unit received other therapeutic strategies. Sec-
ond, only a minority of patients included in the standard
of care had identified contraindications, and therefore we
may have missed the reasons for the non-prescription of
experimental therapies. Subjective, prescriber-based opin-
ions may be a factor explaining these differences. Third,
we could not explain the reasons for increased LOS in pa-
tients treated with hydroxychloroquine or a combination
of hydroxychloroquine and lopinavir/ritonavir (e.g., com-
plications related to COVID-19 or treatment side effects). We formulated hypotheses, but they should be interpreted
cautiously in light of residual confounding, and causal in-
ference was not possible. Overall, there was an indica-
tion bias, with the reason of prescription being associated
with the outcome of interest. Indeed, patients with more
severe COVID-19 were more likely to receive experimen-
tal therapies. We captured severity of COVID-19 with the
mNEWS, but it was used retrospectively and not to de-
cide which experimental treatment should be used. Anoth-
er limitation was that this study was a single-centre study
and it might not be generalisable. Finally, the costs were
underestimated, as they were based on the most recent year
of reference (2018). COVID-19 costs were higher, due to
the use of more expensive equipment. Our cost estimates
should be updated when the cost for 2020 are available. Lopinavir/ritonavir did not show any significant associ-
ation with LOS or in-hospital mortality. This is in line
with recently published findings showing no benefit of
lopinavir/ritonavir on clinical outcomes for patients hospi-
talised with COVID-19 [3]. In addition, these experimental therapies resulted in im-
portant additional healthcare costs. Our findings showed
that the prolonged LOS resulted in 1010.77 additional days
(95% CI 367.57–1408.14) over the study period, which re-
sulted in a total additional cost of US$ 2,492,214 (95%
CI US$ 916,839–3,450,619). Therefore, experimental
COVID-19 therapy costs were not associated with clinical
benefits, but might have resulted in a reduced availability
of hospital beds and a substantial healthcare cost. During
such a pandemic, it is crucial to keep healthcare systems
afloat and every single additional day of hospitalisation
may become detrimental [28]. Limitations of this study Furthermore, we should
keep in mind that the total additional 1010.77 days and
costs of US$ 2,492,214 are underestimated. Indeed, we
only considered hydroxychloroquine and hydroxychloro-
quine combined with lopinavir/ritonavir for which the LOS
was significantly different from zero and thus excluded
lopinavir/ritonavir alone. In addition, the costs were based
on the most recent year of reference (2018) and obviously
real COVID-19 costs were higher. We also excluded some
patients under hydroxychloroquine and hydroxychloro-
quine combined with lopinavir/ritonavir from the analysis. Therefore, our estimate of COVID-19 therapy costs should
be considered to be conservative. Conclusion Our study showed that, in comparison with the standard
of care, hydroxychloroquine or a combination of hydrox-
ychloroquine and lopinavir/ritonavir was associated with
an increased LOS and no significant association with in-
hospital mortality. Prescribing experimental therapies for
COVID-19 was also associated with additional costs (total
additional costs US$ 2,492,214). Healthcare systems in
a pandemic situation experience tremendous pressure and
prolonged LOS and their related costs are key issues that
all stakeholders should consider. cations
nevertheless
received
it.
This
difference
is
important because the toxicity may outweigh potential HCQ
HCQ + LPV/r
Extra LOS (95% CI)
3.75 (1.37–6.12)
4.19 (1.52–5.31)
Internal medicine
Number of patients
78
151
Additional LOS (95% CI)
292.5 (106.86–477.36)
632.69 (229.52–801.81)
Additional cost of LOS (95% CI) in $
751,152 (274,421–1,225,880)
1,624,773 (589,417–2,059,080)
Cost of experimental therapies in $
96
16,954
Geriatrics
Number of patients
15
7
Additional LOS (95% CI)
56.25 (20.55–91.8)
29 (10.64:3 7.17)
Additional cost of LOS (95% CI) in $
64,807 (23,676–105,766)
33,792 (12,059–42,825)
Cost of experimental therapies in $
18
621
Total additional LOS (95% CI)
348.75 (127.41–569.16)
662.02 (240.16–838,98)
Total additional cost in $
816,073 (298,211–1,331,760)
1,676,141 (618,629–2,118,859)
CI = confidence interval; HCQ = hydroxychloroquine; LOS = length of stay; LPV/r = lopinavir/ritonavir
Swiss Medical Weekly · PDF of the online version · www.smw.ch
Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”. No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Page 6 of 9 Swiss Medical Weekly · PDF of the online version · www.smw.ch Page 6 of 9 Page 6 of 9 Original article Swiss Med Wkly. 2020;150:w20446 population reflected the entire population hospitalised for
COVID-19-related reasons, including elderly people, pa-
tients with comorbidities, and patients with contraindica-
tions to therapies. These subgroups are often excluded
from randomised controlled trials and therefore their find-
ings lack external validity [30]. Indeed, trials exclude some
categories of patients, which limits the generalisability of
their results [29]. Other issues related to external validity
include potential better care in all arms and unrepresenta-
tiveness of healthcare professionals involved in the study
[29]. Therefore, randomised controlled trials often provide
information on achievement in the most favourable condi-
tions, and not real-life conditions. benefits, especially among older patients with more preva-
lent cardiac diseases, who may be at higher risk of adverse
effects [26]. A third hypothesis is that prognostic imbalances between
groups have remained. As it was a naturalistic study, resid-
ual confounding that would not have been captured by
available variables in the database might have remained. Even though we used propensity score analyses to address
this methodological flaw, only known confounders could
be included in the study [27]. Therefore, we could not ex-
clude the possibility that the negative association of hy-
droxychloroquine and hydroxychloroquine combined with
lopinavir/ritonavir and LOS was caused by an unknown
confounding factor, including a time-dependent variable
(such as change in recommendations during the study
course). Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”.
No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Acknowledgments g
We thank Dr Laurent Gétaz and Prof. Laurent Kaiser for their support
and early contribution. Findings from routine care studies, such as the present
study, are crucial as a supplement to randomised controlled
trials for the development of a better understanding of
the potential benefits of therapies [29]. Indeed, our study Implications for clinical practice Our study showed that hydroxychloroquine and lopinavir/
ritonavir were not associated with better clinical outcomes
compared with standard of care. We rather found that pa-
tients who received hydroxychloroquine (alone or com-
bined with lopinavir/ritonavir) had a significantly longer
LOS. Prolonged LOS is associated with increased costs
and therefore our study suggests negative issues among pa-
tients as well as for the whole healthcare system. In the
context of COVID-19, these factors are key strategic ele-
ments because healthcare systems are facing tremendous
pressure worldwide. Swiss Medical Weekly · PDF of the online version · www.smw.ch References 1
Salathé M, Althaus CL, Neher R, Stringhini S, Hodcroft E, Fellay J, et
al. COVID-19 epidemic in Switzerland: on the importance of testing,
contact tracing and isolation. Swiss Med Wkly. 2020;150:w20225. doi:
http://dx.doi.org/10.4414/smw.2020.20225. PubMed. 17
Juurlink DN. Safety considerations with chloroquine, hydroxychloro-
quine and azithromycin in the management of SARS-CoV-2 infection. CMAJ. 2020;192(17):E450–3. doi: http://dx.doi.org/10.1503/
cmaj.200528. PubMed. 2
Siemieniuk RAC, Bartoszko JJ, Ge L, Zeraatkar D, Izcovich A, Kum E,
et al. Drug treatments for covid-19: living systematic review and net-
work meta-analysis. BMJ. 2020;370:m2980. doi: http://dx.doi.org/
10.1136/bmj.m2980. PubMed. 18
Guastalegname M, Vallone A. Could Chloroquine /Hydroxychloroquine
Be Harmful in Coronavirus Disease 2019 (COVID-19) Treatment? Clin
Infect Dis. 2020;71(15):888–9. doi: http://dx.doi.org/10.1093/cid/
ciaa321. PubMed. 3
Horby PW, Mafham M, Bell JL, Linsell L, Staplin N, Emberson J, et al.;
RECOVERY Collaborative Group. Lopinavir-ritonavir in patients ad-
mitted to hospital with COVID-19 (RECOVERY): a randomised, con-
trolled, open-label, platform trial. Lancet. 2020;396(10259):1345–52
doi: http://dx.doi.org/10.1016/S0140-6736(20)32013-4. PubMed. 19
Tanne JH. Covid-19: Trump is criticised for again promoting unortho-
dox medical information. BMJ. 2020;370:m3046. doi: http://dx.doi.org/
10.1136/bmj.m3046. PubMed. 4
Horby P, Mafham M, Linsell L, Bell JL, Staplin N, Emberson JR, et al.,
RECOVERY Collaborative Group. Effect of Hydroxychloroquine in
Hospitalized Patients with Covid-19. N Engl J Med. 2020;383(21):2030–40. doi: http://dx.doi.org/10.1056/NEJ-
Moa2022926. PubMed. 20
Yazdany J, Kim AHJ. Use of Hydroxychloroquine and Chloroquine
During the COVID-19 Pandemic: What Every Clinician Should Know. Ann Intern Med. 2020;172(11):754–5. doi: http://dx.doi.org/10.7326/
M20-1334. PubMed. 21
Randomised evaluation of Covid-19 therapy (recovery) https://www.re-
coverytrial.net/files/protocol-archive/recovery-protocol-
v6-0-2020-05-14.pdf/@@download [accessed 2020 July 14]. 5
https://www.recoverytrial.net/news/statement-from-the-chief-investiga-
tors-of-the-randomised-evaluation-of-covid-19-therapy-recovery-trial-
on-hydroxychloroquine-5-june-2020-no-clinical-benefit-from-use-of-hy-
droxychloroquine-in-hospitalised-patients-with-covid-19 [accessed 2020
July 14]. 22
Kim AHJ, Sparks JA, Liew JW, Putman MS, Berenbaum F, Duarte-Gar-
cía A, et al. A Rush to Judgment? Rapid Reporting and Dissemination of
Results and Its Consequences Regarding the Use of Hydroxychloro-
quine for COVID-19. Ann Intern Med. 2020;172(12):819–21. doi:
http://dx.doi.org/10.7326/M20-1223. PubMed. 6
Griffin S. Covid-19: Lopinavir-ritonavir does not benefit hospitalised
patients, UK trial finds. BMJ. 2020;370:m2650. doi: http://dx.doi.org/
10.1136/bmj.m2650. PubMed. 23
Bonow RO, Hernandez AF, Turakhia M. Hydroxychloroquine, Coron-
avirus Disease 2019, and QT Prolongation. JAMA Cardiol. 2020;5(9):986–7. doi: http://dx.doi.org/10.1001/jamacardio.2020.1782. PubMed. 7
https://www.recoverytrial.net/news/no-clinical-benefit-from-use-of-
lopinavir-ritonavir-in-hospitalised-covid-19-patients-studied-in-recovery
[accessed 2020 July 14]. 8
Stringhini S, Wisniak A, Piumatti G, Azman AS, Lauer SA, Baysson H,
et al. Seroprevalence of anti-SARS-CoV-2 IgG antibodies in Geneva,
Switzerland (SEROCoV-POP): a population-based study. Lancet. 2020;396(10247):313–9. doi: http://dx.doi.org/10.1016/
S0140-6736(20)31304-0. PubMed. 24
Mercuro NJ, Yen CF, Shim DJ, Maher TR, McCoy CM, Zimetbaum PJ,
et al. References Risk of QT Interval Prolongation Associated With Use of Hydrox-
ychloroquine With or Without Concomitant Azithromycin Among Hos-
pitalized Patients Testing Positive for Coronavirus Disease 2019
(COVID-19). JAMA Cardiol. 2020;5(9):1036–41. doi: http://dx.doi.org/
10.1001/jamacardio.2020.1834. PubMed. 9
Prokop M, van Everdingen W, van Rees Vellinga T, Quarles van Ufford
H, Stöger L, Beenen L, et al.; COVID-19 Standardized Reporting Work-
ing Group of the Dutch Radiological Society. CO-RADS: A Categorical
CT Assessment Scheme for Patients Suspected of Having
COVID-19-Definition and Evaluation. Radiology. 2020;296(2):E97–104. doi: http://dx.doi.org/10.1148/radi-
ol.2020201473. PubMed. 25
Tang W, Cao Z, Han M, Wang Z, Chen J, Sun W, et al. Hydroxychloro-
quine in patients with mainly mild to moderate coronavirus disease
2019: open label, randomised controlled trial. BMJ. 2020;369:m1849. doi: http://dx.doi.org/10.1136/bmj.m1849. PubMed. 26
Borba MGS, Val FFA, Sampaio VS, Alexandre MAA, Melo GC, Brito
M, et al.; CloroCovid-19 Team. Effect of High vs Low Doses of Chloro-
quine Diphosphate as Adjunctive Therapy for Patients Hospitalized
With Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)
Infection: A Randomized Clinical Trial. JAMA Netw Open. 2020;3(4):e208857. doi: http://dx.doi.org/10.1001/jamanet-
workopen.2020.8857. PubMed. 10
Geleris J, Sun Y, Platt J, Zucker J, Baldwin M, Hripcsak G, et al. Obser-
vational study of hydroxychloroquine in hospitalized patients with
Covid-19. N Engl J Med. 2020;382(25):2411–8. doi: http://dx.doi.org/
10.1056/NEJMoa2012410. PubMed. 11
Kalil AC. Treating COVID-19-Off-Label Drug Use, Compassionate
Use, and Randomized Clinical Trials During Pandemics. JAMA. 2020;323(19):1897–8. doi: http://dx.doi.org/10.1001/jama.2020.4742. PubMed. 27
Straw S, Witte KK. Observational data during the COVID-19 pandemic:
opportunity with uncertainty. Heart. 2020;106(19):1461–2. doi:
http://dx.doi.org/10.1136/heartjnl-2020-317486. PubMed. 12
Windisch O, Zamberg I, Zanella M-C, Gayet-Ageron A, Blondon K,
Schiffer E, et al. Using mHealth to increase the reach of local guidance
to health professionals as part of an institutional response plan to the
COVID-19 outbreak: Usage analysis study. JMIR Mhealth Uhealth. 2020;8(8):e20025. doi: http://dx.doi.org/10.2196/20025. PubMed. 28
Lemos DRQ, D’Angelo SM, Farias LABG, Almeida MM, Gomes RG,
Pinto GP, et al. Health system collapse 45 days after the detection of
COVID-19 in Ceará, Northeast Brazil: a preliminary analysis. Rev Soc
Bras Med Trop. 2020;53:e20200354. doi: http://dx.doi.org/10.1590/
0037-8682-0354-2020. PubMed. 2020;8(8):e20025. doi: http://dx.doi.org/10.2196/20025. PubMed. 13
Swiss DRG. Règles et définitions pour la facturation des cas selon
SwissDRG. Bern, Switzerland: SwissDRG; 2018. 29
Black N. Why we need observational studies to evaluate the effective-
ness of health care. BMJ. 1996;312(7040):1215–8. doi: http://dx.doi.org/
10.1136/bmj.312.7040.1215. PubMed. 14
Liao X, Wang B, Kang Y. Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”.
No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Disclosure statement No financial support and no potential conflict of interest relevant to
this trial were reported. Swiss Medical Weekly · PDF of the online version · www.smw.ch Page 7 of 9 Original article Swiss Med Wkly. 2020;150:w20446 16
Cortegiani A, Ippolito M, Ingoglia G, Iozzo P, Giarratano A, Einav S. Update I. A systematic review on the efficacy and safety of chloroquine/
hydroxychloroquine for COVID-19. J Crit Care. 2020;59:176–90. doi:
http://dx.doi.org/10.1016/j.jcrc.2020.06.019. PubMed. Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”.
No commercial reuse without permission See http://emh ch/en/services/permissions html Swiss Medical Weekly · PDF of the online version · www.smw.ch Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”.
No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”.
No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”.
No commercial reuse without permission See http://emh ch/en/services/permissions html Swiss Medical Weekly · PDF of the online version · www.smw.ch BMI = body mass index; CI = confidence interval; HCQ = hydroxychloroquine; LOS = length of stay); LPV/r = lopinivir/ritonavir; mNEWS = modified National Early Warning Score;
OR = odds ratio * Simple negative binomial regressions (outcome: length of hospitalisation). † Simple logistic regressions (outcome: mortality). Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”.
No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. Published under the copyright license “Attribution – Non-Commercial – No Derivatives 4.0”.
No commercial reuse without permission. See http://emh.ch/en/services/permissions.html. References Novel coronavirus infection during the
2019-2020 epidemic: preparing intensive care units-the experience in
Sichuan Province, China. Intensive Care Med. 2020;46(2):357–60. doi:
http://dx.doi.org/10.1007/s00134-020-05954-2. PubMed. 30
Rothwell PM. External validity of randomised controlled trials: “to
whom do the results of this trial apply?”. Lancet. 2005;365(9453):82–93. doi: http://dx.doi.org/10.1016/
S0140-6736(04)17670-8. PubMed. 15
Joseph BA, Dibas M, Evanson KW, Paranjape G, Vegivinti CTR, Sel-
van PT, et al. Efficacy and safety of lopinavir/ritonavir in the treatment
of COVID-19: A systematic review. Expert Rev Anti Infect Ther. 2020;14787210.2021.1848545. doi: http://dx.doi.org/10.1080/
14787210.2021.1848545. PubMed. Swiss Medical Weekly · PDF of the online version · www.smw.ch Page 8 of 9 Page 8 of 9 Swiss Med Wkly. 2020;150:w20446 Original article Appendix 1: Supplementary table
Table S1: Associations of factors included in the propensity score with outcomes. LOS*
Mortality†
Estimate (95% CI)
p-value
OR (95% CI)
p-value
Gender (ref. female)
0.05 (−0.04 – 0.14)
0.241
20.17 (10.32–30.56)
0.002
Age in years
0.01 (0.003–0.01)
<0.001
10.10 (10.07–10.14)
<0.001
Location (ref. internal medicine)
0.16 (0.04–0.28)
0.012
10.22 (00.67–20.25)
0.516
Month of hospitalisation (ref. Feb./March)
0.12 (0.03–0.21)
0.007
00.39 (00.22–00.67)
0.001
mNEWS
0.03 (0.01–0.05)
<0.001
10.42 (10.28–10.57)
<0.001
Number of diseases
0.09 (0.06–0.13)
<0.001
10.43 (10.19–10.73)
<0.001
BMI ≥30 kg/m2
−0.02 (−0.14 – 0.10)
0.752
10.08 (00.56–20.10)
0.818
Contraindications HCQ
0.15 (0.03–0.26)
0.012
10.01 (00.58–10.77)
0.910
Contraindications LPV/r
0.07 (−0.04 – 0.17)
0.211
10.03 (00.61–10.75)
0.971
BMI = body mass index; CI = confidence interval; HCQ = hydroxychloroquine; LOS = length of stay); LPV/r = lopinivir/ritonavir; mNEWS = modified National Early Warning Score; Appendix 1: Supplementary table BMI = body mass index; CI = confidence interval; HCQ = hydroxychloroquine; LOS = length of stay); LPV/r = lopinivir/ritonavir; mNEWS = modified National Early Warning Score;
OR = odds ratio * Simple negative binomial regressions (outcome: length of hospitalisation). † Simple logistic regressions (outcome: mortality). Swiss Medical Weekly · PDF of the online version · www.smw.ch Page 9 of 9 Page 9 of 9
|
https://openalex.org/W3104341158
|
https://www.epj-conferences.org/articles/epjconf/pdf/2017/04/epjconf_arena2017_01006.pdf
|
English
| null |
Results and Perspectives of the Auger Engineering Radio Array
|
EPJ web of conferences
| 2,017
|
cc-by
| 3,314
|
Results and Perspectives of the Auger Engineering Radio Array
Christian Glaser1,⋆for the Pierre Auger Collaboration2
1RWTH Aachen University, III. Physikalisches Institut A, Aachen, Germany
2Full author list: http://auger.org/archive/authors_2016_06.html Abstract. The Auger Engineering Radio Array (AERA) is an extension of the Pierre
Auger Cosmic-Ray Observatory. It is used to detect radio emission from extensive air
showers with energies beyond 1017 eV in the 30 - 80 MHz frequency band. After three
phases of deployment, AERA now consists of more than 150 autonomous radio stations
with different spacings, covering an area of about 17 km2. It is located at the same site
as other Auger low-energy detector extensions enabling combinations with various other
measurement techniques. The radio array allows different technical schemes to be ex-
plored as well as cross-calibration of our measurements with the established baseline
detectors of the Auger Observatory. We report on the most recent technological develop-
ments and give an overview of the experimental results obtained with AERA. In partic-
ular, we will present the measurement of the radiation energy, i.e., the amount of energy
that is emitted by the air shower in the form of radio emission, and its dependence on the
cosmic-ray energy by comparing with the measurement of the the well-calibrated Auger
surface detector. Furthermore, we outline the relevance of this result for the absolute
calibration of the energy scale of cosmic-ray observatories. © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative
Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). ⋆e-mail: glaser@physik.rwth-aachen.de ,
(2017)
135
EPJ Web of Conferences
ARENA 2016
01006 ,
(2017)
135
EPJ Web of Conferences
ARENA 2016
01006 DOI: 10.1051/
7135
epjconf/201
01006 1 The Auger Engineering Radio Array A
f th
i
t d i Fi
2 Th AERA t ti
i
ith li htl diff
t Figure 1. Pictures of the
two station designs
deployed at AERA, left:
log-periodic dipole
antenna (LPDA), right:
Butterfly antenna. Figure 1. Pictures of the
two station designs
deployed at AERA, left:
log-periodic dipole
antenna (LPDA), right:
Butterfly antenna. The first 24 stations that were installed in 2011 are equipped with log-periodic dipole antennas
(LPDAs). The antenna response to cosmic-ray signals was previously simulated and measured exten-
sively to an accuracy of 12.5% [4, 5]. The stations are placed on a triangular grid with a distance of
144 m covering an area of 0.6 km2 and are connected via optical fibers to the data acquisition. All
other AERA stations are equipped with a Butterfly antenna and were installed in 2013 (100 additional
stations on a 250 m and 375 m grid) and 2015 (29 additional stations on a 750 m grid). Instead of an
optical fiber network the stations are equipped with a wireless communication system operating in the
5 GHz band. All stations operate in the band from 30 to 80 MHz and the radio signal is digitized with
180 MHz or 200 MHz depending on the type of digitizing hardware. The first 24 stations that were installed in 2011 are equipped with log-periodic dipole antennas
(LPDAs). The antenna response to cosmic-ray signals was previously simulated and measured exten-
sively to an accuracy of 12.5% [4, 5]. The stations are placed on a triangular grid with a distance of
144 m covering an area of 0.6 km2 and are connected via optical fibers to the data acquisition. All
other AERA stations are equipped with a Butterfly antenna and were installed in 2013 (100 additional
stations on a 250 m and 375 m grid) and 2015 (29 additional stations on a 750 m grid). Instead of an
optical fiber network the stations are equipped with a wireless communication system operating in the
5 GHz band. All stations operate in the band from 30 to 80 MHz and the radio signal is digitized with
180 MHz or 200 MHz depending on the type of digitizing hardware. A map of the array is presented in Fig. 2. The AERA stations are running with slightly different
types of electronics. 1 The Auger Engineering Radio Array Most of the stations (filled triangles) are equipped with a ring buffer large enough
to store the measured radio signals for 7 s to be able to receive an external trigger from the SD and
FD. The rest of the stations which are equipped with a Butterfly antenna contain small scintillators for
trigger purposes. 1 The Auger Engineering Radio Array The Auger Engineering Radio Array (AERA) is part of the Pierre Auger Observatory [1] and measures
extensive air showers with energies above 1017 eV. The Pierre Auger Observatory is the world’s
largest detector for ultra-high energy cosmic rays. It is located in western Argentina and covers an
area of 3000 km2. Its two baseline detectors are a surface detector (SD) array of more than 1600
water-Cherenkov detectors and 27 fluorescence telescopes (FD) that overlook that array from four
different sites. In addition, underground scintillators are installed at the AERA site that allow for a
separate measurement of the muonic air-shower component. In its current stage of expansion, AERA consists of 153 autonomously operating radio detector
stations covering an area of 17 km2. Pictures of the two station designs deployed at AERA are shown
in Fig. 1. Each station is solar powered and equipped with a battery to run 24 h each day. A fence is
put around each station to protect it from animals living in the Argentinian Pampa and the stations are
equipped with GPS receivers for accurate timing. In addition, a method was develop to calibrate the
relative timing of the radio stations to better than 2 ns using a reference beacon and radio signals from
commercial airplanes [2, 3]. ,
(2017)
135
EPJ Web of Conferences
ARENA 2016
01006 DOI: 10.1051/
7135
epjconf/201
01006 Figure 1. Pictures of the
two station designs
deployed at AERA, left:
log-periodic dipole
antenna (LPDA), right:
Butterfly antenna. The first 24 stations that were installed in 2011 are equipped with log-periodic dipole antennas
(LPDAs). The antenna response to cosmic-ray signals was previously simulated and measured exten-
sively to an accuracy of 12.5% [4, 5]. The stations are placed on a triangular grid with a distance of
144 m covering an area of 0.6 km2 and are connected via optical fibers to the data acquisition. All
other AERA stations are equipped with a Butterfly antenna and were installed in 2013 (100 additional
stations on a 250 m and 375 m grid) and 2015 (29 additional stations on a 750 m grid). Instead of an
optical fiber network the stations are equipped with a wireless communication system operating in the
5 GHz band. All stations operate in the band from 30 to 80 MHz and the radio signal is digitized with
180 MHz or 200 MHz depending on the type of digitizing hardware. 2 Scientific Potential measurement with AERA (see Sec. 4 or [6–8]). Furthermore, the position of the shower maximum
Xmax, which is an estimator of the primary-particle type of the cosmic ray, is directly measured with
the fluorescence telescopes. Coincident measurements of AERA with FD are thus used to evalu-
ate the performance of the different techniques used at AERA to reconstruct the cosmic-ray particle
type. Different methods are developed at present to derive the cosmic-ray coposition from the radio
measurements [9]. measurement with AERA (see Sec. 4 or [6–8]). Furthermore, the position of the shower maximum
Xmax, which is an estimator of the primary-particle type of the cosmic ray, is directly measured with
the fluorescence telescopes. Coincident measurements of AERA with FD are thus used to evalu-
ate the performance of the different techniques used at AERA to reconstruct the cosmic-ray particle
type. Different methods are developed at present to derive the cosmic-ray coposition from the radio
measurements [9]. In addition to a reconstruction of Xmax using solely radio data, the AERA measurement can be
combined with the other detectors. The radio signal measures the electromagnetic component of the
air shower. Hence, a coincident measurement of the muonic shower component using underground
particle detectors is sensitive to the cosmic-ray particle type. In case of horizontal air showers, the
electromagnetic part of the air shower is completely absorbed in the atmosphere so that the SD mea-
sures only the muonic shower component whereas AERA still measures the electromagnetic part of
the shower as the atmosphere is transparent to radio waves. Hence, a combined radio and particle
measurement of horizontal air showers is sensitive to the cosmic-ray particle type [10]. 2 Scientific Potential The different antenna and electronics designs implemented in AERA and the location of AERA next
to additional detectors of the Pierre Auger Observatory where air showers are measured with three
independent detection techniques create a large engineering and scientific potential. AERA can be used to explore the optimal setup to measure cosmic rays. Two different antenna
designs are tested in the harsh environment of the Argentinian Pampa. The different detector spacings
allow the determination of the optimal layout for different energy regimes, e.g., the 750 m spaced
stations are located directly next to SD stations to test the potential of a large-scale extension of
a surface particle detector with radio antennas. The different types of station hardware allow the
implementation of different trigger schemes such as self-triggering directly on the radio signal using
advanced signal processing in an FPGA, external triggering or an internal particle trigger using the
small scintillators integrated into the station design. The latter setup is especially interesting for a
possible future stand-alone radio array. The Auger surface detector, which is calibrated with the fluorescence detector, provides an ac-
curate measurement of the cosmic-ray energy and is used to develop and cross-calibrate the energy 2 2 ,
(2017)
135
EPJ Web of Conferences
ARENA 2016
01006 DOI: 10.1051/
7135
epjconf/201
01006 1 km
AERA24 LPDA, ext. trig. AERA24 LPDA, int. trig. AERA124 butterfly, ext. trig. AERA124 butterfly, int. trig. AERA153 butterfly, ext. trig. central radio station
beacon
FD site
FD FOV
HEAT FOV
SD station
muon-counter
Figure 2. Map of AERA
within the Pierre Auger
Observatory. The radio
detector stations
(triangles) equipped with
different antennas and
digitizing hardware are
surrounded by surface
detector (SD) stations
(gray filled circles) and
underground muon
counters (black
pentagons). AERA is in
the field of view of the
Coihueco and HEAT
fluorescence telescopes. Fig. adapted from [11]. 1 km
AERA24 LPDA, ext. trig. AERA24 LPDA, int. trig. AERA124 butterfly, ext. trig. AERA124 butterfly, int. trig. AERA153 butterfly, ext. trig. central radio station
beacon
FD site
FD FOV
HEAT FOV
SD station
muon-counter
Figure 2. Map of AERA
within the Pierre Auger
Observatory. The radio
detector stations
(triangles) equipped with
different antennas and
digitizing hardware are
surrounded by surface
detector (SD) stations
(gray filled circles) and
underground muon
counters (black
pentagons). AERA is in
the field of view of the
Coihueco and HEAT
fluorescence telescopes. Fig. adapted from [11]. 3 In-situ Calibration of Antenna Response The most challenging part of the detector calibration is the absolute calibration of the antenna response
to cosmic-ray radio signals. To be unaffected by near-field effects, a calibrated signal source needs
to be placed at ∼20 m distance at any place around the antenna. Several calibration campaigns were
performed and the measurement setup was continuously improved. The first campaign used a weather
balloon to lift the calibrated signal source and measured the antenna response with an accuracy of
12.5% [4, 8]. The disadvantages of this method are the high costs of each balloon and the need
for a windless environment. Therefore a new method that uses a remotely piloted drone to lift the
signal source was developed. This setup was completed with an optical camera system to accurately
determine the position of the signal source during the flight. Using this new method an accuracy of
9.3% was achieved for the LPDA antenna [5, 12]. 3 3 ,
(2017)
135
EPJ Web of Conferences
ARENA 2016
01006 DOI: 10.1051/
7135
epjconf/201
01006 ,
(2017)
135
EPJ Web of Conferences
01006 -400
-200
0
200
400
position in ⃗v × ⃗B [m]
-400
-300
-200
-100
0
100
200
300
400
position in ⃗v × (⃗v × ⃗B) [m]
data
sub-threshold
core (SD)
0
10
20
30
40
50
60
70
80
energy fluence f [eV/m2]
1017
1018
1019
ECR [eV]
105
106
107
108
109
E Auger
30−80 MHz/ sin2 α [eV]
A · 107eV(ECR/1018eV)B, A = 1.58 ± 0.07, B = 1.98 ± 0.04
3 - 4 stations with signal
≥5 stations with signal
Figure 3. (left) Energy fluence for an extensive air shower with an energy of 4.4 × 1017 eV and a zenith angle
of 25◦as measured in individual AERA radio detectors (circles filled with color corresponding to the measured
value) and fitted with the azimuthally asymmetric, two-dimensional signal distribution function (background
color). The fit is performed in the plane perpendicular to the shower axis, with the x axis oriented along the
direction of the Lorentz force for charged particles propagating along the shower axis v in the geomagnetic
field B. The best-fitting impact point of the air shower is at the origin of the plot, slightly offset from the one
reconstructed with the Auger surface detector [core (SD)]. (right) Correlation between the normalized radiation
energy and the cosmic-ray energy ECR as determined by the Auger surface detector. 3 In-situ Calibration of Antenna Response Open circles represent air
showers with radio signals detected in three or four radio detectors. Solid circles denote showers with five or
more detected radio signals. Figures and Caption from [7]. -400
-200
0
200
400
position in ⃗v × ⃗B [m]
-400
-300
-200
-100
0
100
200
300
400
position in ⃗v × (⃗v × ⃗B) [m]
data
sub-threshold
core (SD)
0
10
20
30
40
50
60
70
80
energy fluence f [eV/m2]
1017
1018
1019
ECR [eV]
105
106
107
108
109
E Auger
30−80 MHz/ sin2 α [eV]
A · 107eV(ECR/1018eV)B, A = 1.58 ± 0.07, B = 1.98 ± 0.04
3 - 4 stations with signal
≥5 stations with signal Figure 3. (left) Energy fluence for an extensive air shower with an energy of 4.4 × 1017 eV and a zenith angle
of 25◦as measured in individual AERA radio detectors (circles filled with color corresponding to the measured
value) and fitted with the azimuthally asymmetric, two-dimensional signal distribution function (background
color). The fit is performed in the plane perpendicular to the shower axis, with the x axis oriented along the
direction of the Lorentz force for charged particles propagating along the shower axis v in the geomagnetic
field B. The best-fitting impact point of the air shower is at the origin of the plot, slightly offset from the one
reconstructed with the Auger surface detector [core (SD)]. (right) Correlation between the normalized radiation
energy and the cosmic-ray energy ECR as determined by the Auger surface detector. Open circles represent air
showers with radio signals detected in three or four radio detectors. Solid circles denote showers with five or
more detected radio signals. Figures and Caption from [7]. 4 Measurement of the Radiation Energy of Extensive Air Shower To measure the radiation energy, i.e., the amount of energy that is transferred from the cosmic ray
during the air shower development into radio emission in the frequency band of AERA (30 - 80 MHz),
we first unfold the detector response from the measured voltage traces. Thereby we obtain the electric
field of the cosmic-ray radio signal from which we calculate the energy fluence, i.e., the energy in the
radio signal per unit area, at each radio station. A two-dimensional function [8, 13] is fitted to the
signal distribution and integrated to obtain the radiation energy of the air shower (cf. Fig. 3 left). We correct the radiation energy for the different emission strengths at different geomagnetic angles
α by dividing the radiation energy by sin2 α. This corrected radiation energy correlates with the energy
of the cosmic ray. Comparing the radiation energy with a measurement of the cosmic-ray energy by
the surface detector we found that the radiation energy scales quadratically with the cosmic-ray energy,
and we measured a radiation energy of 15.8±0.7(stat)±6.7(sys) MeV for a 1 EeV air shower arriving
perpendicular to the geomagnetic field (cf. Fig. 3 right). We can generalize our measurement to be applicable at any place on Earth by normalizing the
radiation energy to the geomagnetic field strength. Then this corrected radiation energy is the same at
any experiment and our result can be used by others to calibrate against the accurate energy scale of
the Pierre Auger Observatory. More details can be found in [6–8]. 4 ,
(2017)
135
EPJ Web of Conferences
ARENA 2016
01006 DOI: 10.1051/
7135
epjconf/201
01006 6 Conclusions The Auger Engineering Radio Array is the world’s largest radio detector for high-energy cosmic rays
consisting of 153 autonomously-operating radio stations covering an area of 17 km2. Its location
within the Pierre Auger Observatory creates a large scientific potential as air showers are measured
with four independent detection techniques simultaneously. The radiation energy of air showers was
measured and compared with the accurate cosmic-ray energy information of the Auger surface de-
tector. Furthermore, the radiation energy can be calculated from first principles for an independent
determination of the cosmic-ray energy scale. 5 Independent Determination of Cosmic-Ray Energy Scale from First
Principles The measurement of the radiation energy can also be used for an independent determination of the
cosmic-ray energy scale. The radiation energy released by air showers can be calculated from first
principles. The radio emission originates only from the well-understood electromagnetic part of the
air shower and is due to (de)acceleration of charges which is described by classical electrodynamics. Such a calculation is implemented in modern Monte-Carlo simulations [14, 15] which can be used for
an accurate prediction of the radiation energy [7, 16, 17]. Compared to a measurement of fluorescence light, the measurement of radiation energy has the
advantage that it is hardly influenced by environmental conditions. The atmosphere is transparent to
radio waves in the frequency range of AERA and also the amount of radiation energy released by an
air showers is barely influenced by the current state of the atmosphere [16]. References [1] A. Aab et al. (Pierre Auger Collaboration), Nucl. Instrum. Meth. A 798, 172 (2015) [1] A. Aab et al. (Pierre Auger Collaboration), Nucl. Instrum. Meth. A 798, 172 (2015) [2] A. Aab et al. (Pierre Auger Collaboration), J. Inst. 11, P01018 (2016) [3] T. Huege for the Pierre Auger Collaboration, this conference (2016) [4] P. Abreu et al. (Pierre Auger Collaboration), J. Instrum. 7, P10011 (2012) [5] R. Krause for the Pierre Auger Collaboration, Nucl. Instrum. Meth. A, Proc. 14th, Vienna Con-
ference on Instrumentation (VCI), Vienna, Austria (2016) [6] C. Glaser for the Pierre Auger Collaboration, Proceedings of Science (2015), 34th ICRC, The
Hague, The Netherlands [7] A. Aab et al. (Pierre Auger Collaboration), Phys. Rev. Lett. 116, 241101 (2016), 160 [ ]
(
g
),
y
,
(
),
[8] A. Aab et al. (Pierre Auger Collaboration), Phys. Rev. D 93, 122005 (2016), 1508.04267 [8] A. Aab et al. (Pierre Auger Collaboration), Phys. Rev. D 93, 122005 (2016), 1508 [9] F. Gaté for the Pierre Auger Collaboration, this conference (2016) [10] O. Kambeitz for the Pierre Auger Collaboration, this conference (2016) [11] J. Schulz for the Pierre Auger Collaboration, Proceedings of Science (2015), 34th ICRC, The
Hague, The Netherlands [12] F. Briechle for the Pierre Auger Collaboration, this conference (2016) [13] A. Nelles et al., Astropart. Phys. 60, 13 (2015) [14] J. Alvarez-Muñiz, W.R. Carvalho, E. Zas, Astropart. Phys. 35, 325 (2012), 1107. [15] T. Huege, M. Ludwig, C.W. James, AIP Conf. Proc. 1535, 128 (2013) [16] C. Glaser, M. Erdmann, J.R. Hörandel, T. Huege, J. Schulz, submitted to JCAP (2016),
1606.01641 [17] C. Glaser, M. Erdmann, J.R. Hörandel, T. Huege, J. Schulz, this conference (2016) 5 5
|
https://openalex.org/W2011706112
|
https://zenodo.org/records/1583463/files/article.pdf
|
English
| null |
Sermons in Stomach Stones
|
Science
| 1,906
|
public-domain
| 3,745
|
SCIENCE. SCIENCE. the sections entitled by him 'Etude de la
p2te fgcale,' and 'R&sum6 de quelques ex-
periences sur la destruction de divers types
de crottins,' the latter inciuding a notable
category. Our foreign colleague is conspicu-
ous for his firm grasp of the subject-matter,
and his ornate handling of it sets an example
which might well be emulated by his brethren
on this side of the water. But then, as Seneca
observes, speaking with all due respect:
'Gallurn in sw, sterquilinio plurirnurn posse.'
C. R. EASTMAN. SERMONS IN STOMACII STONES. MOLI~RE
presents to us in one of his come-
dies the father of a dumb girl who wants to
know why his daughter is dumb. 'Nothing
is more easy to explain,' says the pretended
physician Sganarelle; 'it comes from her
having lost the power of speech.'
'Yes, yes,'
objects the father, 'but the cause, if you
please, why she has lost the power of speech? '
Sganarelle is at no loss for an answer: 'All
our best authors will tell you that it is the
impeding action of the tongue.' p
p
C. R. EASTMAN Sornewhat similarly, if one asks the defini-
tion of a 'stomach stone,' one is told that it
is a 'gastrolith ';' and our best authors de-
clare sapiently that gastroliths are pebbles
that have been swallowed by fossil reptiles of
'lithophagous proclivities.'
Predilection for
this hard fare is accounted for by ascribing
to the reptiles in question a bird-like gizzard;
and the upshot of the matter is that we find
evidence 'of additional important structural
analogies with the birds,' quod erat invenien-
durn. Huch the same method of reasoning
led the jovial Tom Hood, in his 'Geological
Exdursion to Tilgate Forest, A.D. 2000,' to
affirm that Mylodon, subsisted upon a diet of
'raw potatoes and undressed salads.' 'SCIENCE,V01. XXIJI., p. 820.
a Ibid., Vol. SX., p. 565.
'Les coprolithes de Rernissart.'
Mem. Musee
Roy. d'Hist. Nat. Belg., T. J., 1903. 'SCIENCE,V01. XXIJI., p. 820.
a Ibid., Vol. SX., p. 565. 'Les coprolithes de Rernissart.'
Mem. Musee
Roy. d'Hist. Nat. Belg., T. J., 1903. 'SCIENCE,V01. XXIJI., p. 820.
a Ibid., Vol. SX., p. 565.
' TI-IE FUTURE OB' THE CRAYFISH INDUSTRY. central cities belong to the American genus
Cambarus, which occurs in the United States
and Canada only. The Potomac supplies C. afinis; Chicago, C. airilis; New Orleans, C. Ulandinyii; and Montreal, C. Bartoni. Of
late, however, a considerable fishery has de-
veloped in the Pacific states where the cray-
fish are all of the genus Asiacus and more like
thc crayfish of England, France and Europe
in general. From the statistics of the Bureau
of Fisheries we learn that in Oregon 116,400
pounds of crayfish, worth $7,760, were caught
in 1899. The detailed tables, however, as-
sign 63,000 pounds, worth $420, to Clackamas
County, where they are talren in the tributaries
of the Willamette River along a stretch of
only a few ruiles; 5,400 pounds, worth $360, to
Columbia
County; 15,000 pounds, worth
$1,000, to Multnomah County; 15,000 pounds,
worth $1,000, to Washington County; and
18,000 pounds, worth $1,200, to Yamhill
County. This would make a total of 165,000
pounds in place of 116,400 pounds. The future of this crayfish industry will
obviously depend upon both demand and sup-
ply. The demand should increase; with the
growth of cosmopolitan populations that ap-
preciate such food as is used in Europe; with
the growth of large populations too remote
from sea coast to obtain fresh sea food; and
with the increasing inadequacy of the marine
crustacea to supply the needs of even those
consumers who dwcll near the coast. Thus
the lobster industry has been straincd till the
use of young specimens as food to take the
placc of the exterminated large oncs has be-
cotne very extensive. At prcscnt sorne mil-
lions of 'short-lobster's,' six to ten inches long,
are sold to summer visitors to the New Eng-
land coast and many more millions are used
as bait. No doubt, in time, the demand for crayfish
will exceed the natural supply .and this in-
dustry will tend to run the same retrograde
course as that of the lobster, oyster, clam and
many more important fisheries till the real,
or assumed, value of the crayfish as food, war-
rants legislakive cowtrol and scientific aid
such as alone makes possible the continuance
of more and more of our once 'inexhaustible '
food supplies. TI-IE FUTURE OB' THE CRAYFISH INDUSTRY. CRAYFISHin the United States form so small
a part of the food supply that we are apt to
rank them with mussels and snails as eaten in
Europe only. But while France so highly,
appreciates them as to carry on the business
of rearing them to increase the natural supply
coming from her own waters and those of
neighboring countries ;there is actually a grow-
ing consumption of crayfish as food in the
TJnited States. In New York, New Orleans,
San Francisco, Chicago and other cities cray-
fish are sold both as food and as garnish, as
bait and as material for school and college
courses in zoology. We have no wish to impugn the worth of
stomach stones, nor of related bodies known
as uroliths and coprolites, as a fit subject for
scientific inquirsjand as a means of satisfy-
ing hunger and &iwt after knowledge. All
are capable of large returns, as witness, for
example, the fecund results of M. Bertrand,
whose memoir3 of 150 odd pages, illustrated
by fifteen plates, is at once edifying, delect-
able and digne; that is, fully commensurate
with the ,materials. The argumentation em-
ployed is informed with severest logic, in
which undisciplined imagination has no place;
and the author seeks to test, verify, or at least
fortify, his conclusions as far as possible by
the experimental method. An evident Iong-
ing to get at the bottom of things is shown in While the actual status of the crayfish in-
dustry is difficult to determine, the following
facts show that if a complete census were
taken it would show the existence of a much
larger use of crayfish than is at all suspected. One small region, the Potomac from Wash-
ington to Fort Washington, was recently esti-
mated by one of the most intelligent fishermen
on the Maryland side to send annually to New
York a half million of crayfish, while the U. S. Fish Commission publications in 1884 asserted
that Montreal and Milwaukee also shipped
cra6sh to New York. More recent reports
of the commission state that in 1902 the cray-
fish catch in New Orleans County, Louisiana,
was 16,000 pounds, of a value of $615, and of
Monroe County, Florida, 55,664 pounds, of a
value of $3,282. All the above crayfish and many more
caught for the markets of Chicago and other SCIENCE. [N. S. VOL.XXIII. No. GOO. l'Notes on Fisheries of the Pacific Coast in
1899,' p. 545. TI-IE FUTURE OB' THE CRAYFISH INDUSTRY. This large species has been sold for twice the
price of the eastern or the southern crayfishes,
and basides its larger size and weight it has
the advantage of more attractive and lobster-
like appearance, so that its introduction into
the east should be most acceptable. In fact,
large specimens brought here and kept alive
in the laboratory were as long as the six-inch
'short lobster ' now used as food and if these
crayfish were available in qua
be used as a substitute for suc
and thus protect the lobster
As the scientific study of
distribution of crayfish leads
sion that the Oregon crayhh
sent from the whole eastern a
because its ancestors never g
because of any natural obsta
there when once introduced,
well worth the while for the
eries, and for private individu
large numbers of young an
into waters near markets no
with this superior article of f
be found that this large Asta
clirnated in the east to com
barz~,say in the Potomae, o
the vacant place4 not now o
crayfish at all (such as the Co
the catching of such introdu
be a lucrative business that w
ceptable article to the food d
waters. TI-IE FUTURE OB' THE CRAYFISH INDUSTRY. As each male is capable of fertilizing several
females and, moreover, as it was found that
in Cambarus the sperm may be kept all winter
in the receptacle of the female and used by
her to fertilize the eggs in the spring, few
males are needed for breeding and these could
be used as food in tho winter before the
spring, when so many of them die. This kind of erasfish grew in the laboratory
to a length of four inches in three and a
quarter years and was of marlretable size,
three inches, at the end of the second summer
from the egg. Probably in the open it at-
tains its maximum size in four or five years. A third means of increasing the available
food supply-the
origination of larger races
of crayfish-may
remain for a later stage of
the industry, but as we have more than sixty
species of Cambarus besides several species of
-4stacus in this country and nine or more other
genera in other countries, the chances would
seem good for some future production of new
forms from crossing and selection. y
A second method of improving the supply
of crayfish, the introduction of other species,
seems a promising field for experiment, for it
has been found here that the eggs of the large
Oregop crayfish, Astacus, may be hatched in
the laboratory and reared as readily as in the
case of C. afinis, the native species. These
young Oregon crayfish grew here under such
artificial conditions to a length of 60 mm. in
five months from eggs hatched in the spring. This large species has been sold for twice the
price of the eastern or the southern crayfishes,
and basides its larger size and weight it has
the advantage of more attractive and lobster-
like appearance, so that its introduction into
the east should be most acceptable. TI-IE FUTURE OB' THE CRAYFISH INDUSTRY. The center of the wholesale crayfish busi-
ness was Portland, in Multnomah County,
where the sales were 39,232 dozen crayfish,
weighing 117,696 pounds and worth $19,556.' The catch is made in the sloughs of the Colum-
bia. and its tributary streams between March and
September. A large part of the catch is used
at Portland, with a considerable $emand from
Seattle, Tacoma, San Francisco and as far east
as Salt Lake City and St. Louis. The average
weight is three pounds to the dozen. As pre-
pared for rrhipment the crawfish is placed alive
in a composition of white wine and spices and
boiled for about ten minutes. The crawfish and
liquor in which it has been boiled are next
packed in tin buckets holding from two to three
dozen each. Soon or later the supply of crayfish will
need to be made greater. In addition to
legislative
restrictions and controls three
lines of work suggest themselves as suitable
for trial when the supply becomes deficient or,
if one is to profit by experience in other fish-
eries, now, before the supply is deficient. First the artificial breeding of native species
in the market region; second, the introduction
and propagation of better species than those
naturally occurring; and thirdly, the improve-
ment in size and flavor by culture and cross-
breeding. Despite the incompleteness of the above
data it is evident that considerable numbers
of crayfish are sold and that they 5nd a market
even in Pacific, Atlantic and Gulf states,
where they compete with salt-water crustacea;
in San Francisco with the spiny-lobster and
crab, in New Orleans with the shrimp and
in New York with the lobster. Crayfish amongst
crustacea,
like carp
amongst fish, lend then~selves readily to pond
culture and breeding. Experiments carried on
here in the laboratory have demonstrated the
ease with which the young of C. aftinis can be
reared and have shown two facts of econoinic
value, namely, that the young reared from
eggs laid in the spring may become sexually l'Notes on Fisheries of the Pacific Coast in
1899,' p. 545. 'NCE. crayfish were available in quantity they might
be used as a substitute for such young lobsters
and thus protect the lobster industry. mature and lay eggs the following spring
when not quite one year old, and also that a
female may lay eggs in two successive years. TI-IE FUTURE OB' THE CRAYFISH INDUSTRY. Even the artificial
larger crayfish in central an
awaits but the developments
profitable side of fresh-water
A third means of increasi
food supply-the
origination
of crayfish-may
remain for
the industry, but as we have
species of Cambarus besides s
-4stacus in this country and n
genera in other countries, th
seem good for some future pr
forms from crossing and sele
In France as far back as 1
crayfish farm supplied Leon
the data for making out the
Astacus, and as we have fou
Gambarus here so similar the
stacle to the establishment of
the United States except the
cient demand for crayfish as
The needs of both Astacu
have been shown in this l
simply air, a small amount of
organic food which may be a
nzimal and vegetable matter As the scientific study of the geographical
distribution of crayfish leads to the conclu-
sion that the Oregon crayhh, Astacus, is ab;
sent from the whole eastern and central states
because its ancestors never got there and not
because of any natural obstacle to its living
there when once introduced, it would seem
well worth the while for the Bureau of Fish-
eries, and for private individuals, to introduce
large numbers of young and adult Astacus
into waters near markets not now supplied
with this superior article of food. Should it
be found that this large Astacus may be ac-
clirnated in the east to compete with Cam-
barz~,say in the Potomae, or better to take
the vacant place4 not now occupied by any
crayfish at all (such as the Connecticut River)
the catching of such introduced forms would
be a lucrative business that would add an ac-
ceptable article to the food drawn from fresh
waters. Even the artificial rearing of these
larger crayfish in central and eastern waters
awaits but the developments of time to be a
profitable side of fresh-water farming. Though the eggs are best cared for by the
mother, it was found possible to hatch them
in McDonald fish-hatching jars and thus rear
them under artificial conditions from a very
early stage. TI-IE FUTURE OB' THE CRAYFISH INDUSTRY. As each lays from two hundred to six hun-
dred eggs, a few breeding females would fur-
nish a large stock of young. T h e young r e
spond readily to feeding and the resulting size
seems as much dependent upon the food as
upon the age of the individual. Large indi-
viduals and large races might be expected
from proper culture. mature and lay eggs the following spring
when not quite one year old, and also that a
female may lay eggs in two successive years. As each lays from two hundred to six hun-
dred eggs, a few breeding females would fur-
nish a large stock of young. T h e young r e
spond readily to feeding and the resulting size
seems as much dependent upon the food as
upon the age of the individual. Large indi-
viduals and large races might be expected
from proper culture. Though the eggs are best cared for by the
mother, it was found possible to hatch them
in McDonald fish-hatching jars and thus rear
them under artificial conditions from a very
early stage. As each male is capable of fertilizing several
females and, moreover, as it was found that
in Cambarus the sperm may be kept all winter
in the receptacle of the female and used by
her to fertilize the eggs in the spring, few
males are needed for breeding and these could
be used as food in tho winter before the
spring, when so many of them die. This kind of erasfish grew in the laboratory
to a length of four inches in three and a
quarter years and was of marlretable size,
three inches, at the end of the second summer
from the egg. Probably in the open it at-
tains its maximum size in four or five years. A second method of improving the supply
of crayfish, the introduction of other species,
seems a promising field for experiment, for it
has been found here that the eggs of the large
Oregop crayfish, Astacus, may be hatched in
the laboratory and reared as readily as in the
case of C. afinis, the native species. These
young Oregon crayfish grew here under such
artificial conditions to a length of 60 mm. in
five months from eggs hatched in the spring. TWO LETTERS OF DR. DARWIN: TBE EARLY
DATE OP HZX EVOLUTIONAL WRZTINGX. SEVERAL
of
have
letters
Erasmus Darwin
lately come into my possession, and two of
them seem worthy of publication, if only for
the reason that reference t o his evolutional
ideas seldom occur in his correspondence. In
this regard, for example, Charles Darwin
states in his introduction to Dr. Krause's
' Erasmus Darwin,' that 'most of the letters
[of his grandfather] which he possessed or
had seen, are uninteresting and not worth
publication.' This however I have now laid aside, for perhaps
more important, tho' less ingenious occupations;
& shall therefore decline giving you the trouble
of sending me the boolis yon are so kind as to
offer, both in your last, 8: in a former letter of
yours, I am Sr. The earlier letter, I may note, has the merit
of referring to Dr. Darwin's work on the
anatomy of plants, and to his ingenious effort
to show closer correspondence between the
organs of the higher plants and the higher
animals. Indeed, as we know froin other
sources, he even expected ultimately to find
i n plants the homologues of the animal nerves,
ganglia and sense organs. Accordingly, we
are not surprised to find that he refers here,
in quite a matter of fact wag, to the 'blood'
and the 'two systems ' of a plant. And he
gives us also a glimpse of laboratory methods,
and of his interest in getting in prompt touch
with the results of foreign workers. with great respect your obedt. servtt. E. DARWIN The second letter is of livelier interest. H e
denies having ' stolen ' his ' Botanic Garden,'
or of even having heard of its prototype, prob-
ably the ' Universal Beauty ' of Brooke (1735). And he modestly predicts of the effect of his
evolutional ' conjectures.'
Finally, he refers
to the ' Zoonomia,' as having been on his work
table--or
rather 'lain by him' for 'nearly
twenty y e a r s ' i . e., since about 1771. BALTIMORE,May, 1906. BALTIMORE,May, 1906. On looking over Malpigl~i,& Grew, & I'lales,
the physiology of plants appear'd to me, not t o
have hitherto been under the attention of any one
perfectly acclllaintccl with the anirnal economy. Last summer I contrived to inject the absorbent
systcm of the Pier is with a colour'd liquor; & as
the blood of that plant is white, these two sys-
tems were beautifully apparent to the eye. On
reading a manuscript translation of Mr. -
a Sweedish naturalist, I found the authors, I
mentioned to you in my last, had made a set of
similar experiments; & I had designed t o have in-
vestigated this subject, so little understood a t
present, farther during the summer. TWO LETTERS OF DR. DARWIN: TBE EARLY
DATE OP HZX EVOLUTIONAL WRZTINGX. That
the work here referred t o is the 'Zoonomia,'
there can be no doubt; he obviously means a n
extended evolutional work, and, i n a letter to
a son, dated the following year (cited in the
introduction to 'Erasmus Darwin,'
above re-
ferred to, page 102), he says, that "he is
studying his ' Zoonomia.' " It is, of course,
well known that this work was long intended
for posthumous publication. But the exact The first of these letters is addressed to
'Sir Joseph Banks, Barf Soho Square Lon-
don.' and is as follows: RADBUBNMAR. 16-82 Dear Xir, TI-IE FUTURE OB' THE CRAYFISH INDUSTRY. In fact,
large specimens brought here and kept alive
in the laboratory were as long as the six-inch
'short-lobster ' now used as food, and if these In France as far back as 1865 a successful
crayfish farm supplied Leon Soubeiran with
the data for making out the life history of
Astacus, and as we have found the habits of
Gambarus here so similar there seems no ob-
stacle to the establishment of crayfish farms in
the United States except the lack of a suffi-
cient demand for crayfish as food. The needs of both Astacus and Cambarus
have been shown in this laboratory to be
simply air, a small amount of fresh water and
organic food which may be a variety of refuse
nzimal and vegetable matter. The artificial SCIENCE. [N. 8. VOL. XXIII. No. 600. [N. 8. VOL. XXIII. No. 600. abroad, & thence meant to have returncd them
together. culture of crayfish will be profitable as soon as
the market price is greater than the small
cost of food, the inexpensive farm and the
value of the little labor involved. The intro-
duction of the large Oregon crayfish with its
attractive colors and large claws might con-
ceivably so stimulate the general demand as
soon to raise the market value to such a profit-
able level. Mrs. Blacburn favor'd us with a copy of Murray,
but desired it to be returned in three months,
which it was to a day; & as I could procure but
one other, & our society was not all resident at
Lichfield, we were distressed on this account, but
are still flatter'd with daily hopes of Inore copies
being imported. I am sorry you say the re-
mainder of the supplementum is not likely soon
to be had. E. A. ANDREW Dear Xir, I return'd your sixth volum of the Ameenit. academ. & thank you for the loan of it. I should
have sdoner sent it, but hoped to have received
another copy of Murray, & also that Dr. Linneus's
supplementlim would have been procured from
|
https://openalex.org/W2849027590
|
https://www.mdpi.com/2073-4344/8/7/278/pdf?version=1532407906
|
English
| null |
Immobilization of an Endo-β-N-acetylglucosaminidase for the Release of Bioactive N-glycans
|
Catalysts
| 2,018
|
cc-by
| 12,395
|
Received: 1 June 2018; Accepted: 5 July 2018; Published: 10 July 2018 Abstract: As more is learned about glycoproteins’ roles in human health and disease, the biological
functionalities of N-linked glycans are becoming more relevant. Protein deglycosylation allows for the
selective release of N-glycans and facilitates glycoproteomic investigation into their roles as prebiotics
or anti-pathogenic factors. To increase throughput and enzyme reusability, this work evaluated
several immobilization methods for an endo-β-N-acetylglucosaminidase recently discovered from
the commensal Bifidobacterium infantis. Ribonuclease B was used as a model glycoprotein to compare
N-glycans released by the free and immobilized enzyme. Amino-based covalent method showed
the highest enzyme immobilization. Relative abundance of N-glycans and enzyme activity were
determined using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Kinetic evaluation demonstrated that upon immobilization, both Vmax and the Km decreased. Optimal
pH values of 5 and 7 were identified for the free and immobilized enzyme, respectively. Although a
higher temperature (65 vs. 45 ◦C) favored rapid glycan release, the immobilized enzyme retained over
50% of its original activity after seven use cycles at 45 ◦C. In view of future applications in the dairy
industry, we investigated the ability of this enzyme to deglycosylate whey proteins. The immobilized
enzyme released a higher abundance of neutral glycans from whey proteins, while the free enzyme
released more sialylated glycans, determined by nano-LC Chip Q-ToF MS. Keywords: N-glycans; mass spectrometry; immobilization; prebiotic; glycosidase; recombinant; kinetic;
nano-LC Chip Q-ToF MS Immobilization of an Endo-β-N-
acetylglucosaminidase for the Release of
Bioactive N-glycans Joshua L. Cohen 1, Sercan Karav 2, Daniela Barile 1,3 ID and Juliana M. L. N. de Moura Bell 1,4,*
1
Department of Food Science and Technology, University of California, One Shields Avenue, Davis, CA 95616,
USA; jlcohen@ucdavis.edu (J.L.C.); dbarile@ucdavis.edu (D.B.) 1
Department of Food Science and Technology, University of California, One Shields Avenue, Davis, CA 95616,
USA; jlcohen@ucdavis.edu (J.L.C.); dbarile@ucdavis.edu (D.B.) j
(J
)
(
)
2
Department of Molecular Biology and Genetics, Canakkale Onsekiz Mart University, 17100 Canakkale,
Turkey; sercankarav@comu.edu.tr 3
Foods for Health Institute, University of California, One Shields Avenue, Davis, CA 95616, USA
4
Department of Biological and Agricultural Engineering, University of California, One Shields Avenue,
Davis, CA 95616, USA 3
Foods for Health Institute, University of California, One Shields Avenue, Davis, CA 95616, USA
4
Department of Biological and Agricultural Engineering, University of California, One Shields Avenue,
Davis, CA 95616, USA *
Correspondence: jdemourabell@ucdavis.edu; Tel.: +1-530-752-5007 catalysts catalysts catalysts www.mdpi.com/journal/catalysts Keywords: N-glycans; mass spectrometry; immobilization; prebiotic; glycosidase; recombinant; kinetic;
nano-LC Chip Q-ToF MS 1. Introduction Glycoproteins are a biologically important class of components with modulatory roles in signaling
and cell adhesion. N-linked glycans are covalently bound to an asparagine residue with the consensus
sequence on the primary structure of asparagine-X-serine/threonine (with X representing any amino
acid besides proline) [1]. The attachment is mediated by the reducing end N-acetylglucosamine moiety
of the glycan onto the asparagine residue [2]. All N-glycans share a common trimannosyl, chitobiose
(two β-1,4 linked N-acetylglucosamine residues) core and are classified based on how the core is
elongated with various monosaccharides. N-glycans can be decorated with N-acetylneuraminic acid or
N-glycolylneuraminic acid (sialylated) and/or fucose (fucosylated) in various antennary combinations
to give rise to a heterogeneity of combinations, even on a single glycosylation site. Glycans can Catalysts 2018, 8, 278; doi:10.3390/catal8070278 www.mdpi.com/journal/catalysts Catalysts 2018, 8, 278 2 of 15 be classified based on their antennary decorations, with neutral, fucosylated, sialylated, and both
fucosylated and sialylated being the primary non-plant glycans. be classified based on their antennary decorations, with neutral, fucosylated, sialylated, and both
fucosylated and sialylated being the primary non-plant glycans. As a model for understanding the interplay of indigestible carbohydrates and beneficial gut
bacteria, human milk oligosaccharides (HMOs) have been widely studied and found to exhibit uniquely
effective prebiotic functionalities [3]. While HMOs are quite abundant in human milk (up to 15 g/L in
colostrum), their commercial-scale isolation from human milk is not feasible [4]. The activities and
roles of intact and released N-linked glycans upon human consumption are not well understood due,
in part, to the lack of adequate deglycosylation and analytic methods to release, identify, and quantity
the released glycans [5]. N-glycans can be released from the protein moiety using harsh chemical
treatments or enzymatic methods, the latter allowing for the recovery of intact protein and glycans. Deglycosylating enzymes include peptide-N-glycosidases and endo-β-N-acetylglucosaminidases,
which differ in cleavage points of the core of N-glycans [6]. Due to striking similarities in structures and functions, released N-linked glycans from bovine milk
glycoproteins have been studied for their ability to promote the growth of commensal bacteria in vitro [7,8]. A recently discovered endo-β-N-acetylglucosaminidase (Endo-BI-1), isolated from B. infantis, was shown
to deglycosylate human milk glycoproteins more effectively than other endo-β-N-acetylglucosaminidase
isolated from several commensal Bifidobacterium species [9]. More importantly, released N-glycans by
this novel enzyme from bovine milk proteins displayed a remarkable selective prebiotic activity on
Bifidobacterium longum subsp. 1. Introduction infantis in vitro, and may promote the growth of other beneficial bacteria
while inhibiting pathogens [7,8,10]. However, to establish N-glycans as an alternative source of prebiotic
carbohydrates to human milk, further research into the selectivity of bovine N-glycans and their functional
similarity with HMOs with respect to commensal, beneficial, and pathogenic gut organisms is necessary. Whey, the co-product of cheese manufacture, is a potential commercially available source of
underutilized glycoproteins. The concentration of proteins in bovine cheese whey ranges from 1 to
2%, wherein approximately 4–9% of those proteins are N-glycosylated immunoglobulins, lactoferrin,
and transferrin [11,12]. Lactoferrin, while found in substantial quantities in human milk and bovine
colostrum, is present in bovine milk and cheese whey at the trace level. Yet, the nearly 200 million tonnes of whey produced each year globally [13] conservatively
translates to 2 million tonnes of whey proteins, corresponding to approximately 100,000 tonnes of
bovine milk glycoproteins available for subsequent processing. With such a large availability of
glycoproteins, the development of large-scale processing strategies to release and isolate N-glycans
for further characterization and elucidation of their biological functionalities becomes a key step for
further commercialization of these compounds. Indeed, enzyme immobilization may facilitate reaction scale-up considering the possibility for broad
reactor systems and catalyst reusability. Enzyme immobilization is the process wherein a soluble enzyme
is attached or adsorbed onto a solid support, entrapped within a matrix, or otherwise aggregated
enabling enzyme reuse during production [14]. However, widespread industrial use of immobilized
enzyme remains limited due to a perceived loss of enzyme activity, lack of universal immobilization
techniques, and cost implications [15,16]. In general, enzyme immobilization facilitates the separation
of products and catalyst where removal from the final product to terminate the reaction and to reuse
the catalyst is important in producing pure bioactive molecules. Covalent, adsorption, entrapment,
and aggregation methods of immobilization have their inherent advantages and disadvantages in a
reactor system. These factors are based on substrate size and ability to access the enzyme active site,
physicochemical stability, and applicability in industrial processes [15]. Although the use of immobilized
enzymes has been validated by the food industry to a limited extent [17], the use of glycosidases has been
restricted to lab-scale [6]. The overall goal of this study was to evaluate several types of immobilization methods and resins
for a novel endo-β-N-acetylglucosaminidase (Endo-BI-1) isolated from B. infantis. 2.1. Protein Immobilization Yield
2.1. Protein Immobilization Yield Enzyme immobilization yield of amino-, epoxy- and adsorption-immobilized Endo BI-1 was
evaluated by quantifying the unbound protein remaining in the supernatant and resin washing
buffer post-immobilization. The fluorescence method employed herein (Qubit Protein Assay Kit)
demonstrated the high immobilization yield (>98%) of the amino-based resin, whereas divinylbenzene
(DVB)-based hydrophobic interaction adsorption immobilized 70% of the enzyme (Figure 1). Epoxy-based covalent binding yielded the lowest immobilization yield at 12%. These results were
corroborated by SDS-PAGE, where denser Endo BI-1 bands in the supernatants were present in
epoxy and adsorption methods, while amino-based immobilization had no visible band (Figure S1). Differences between immobilization yields for covalent and adsorption immobilization methods
have been observed previously and can possibly be attributed to the strength of interaction between
hydrophobic interactions and covalent bonds [18]. However, low binding using epoxide-activated
resin may be due to non-optimal methods to identify the ideal pH, time, and temperature for increased
binding [19]. Further investigation into those conditions may improve protein immobilization yield
for the epoxide-activated resin. Enzyme immobilization yield of amino-, epoxy- and adsorption-immobilized Endo BI-1 was
evaluated by quantifying the unbound protein remaining in the supernatant and resin washing
buffer post-immobilization. The fluorescence method employed herein (Qubit Protein Assay Kit)
demonstrated the high immobilization yield (>98%) of the amino-based resin, whereas
divinylbenzene (DVB)-based hydrophobic interaction adsorption immobilized 70% of the enzyme
(Figure 1). Epoxy-based covalent binding yielded the lowest immobilization yield at 12%. These
results were corroborated by SDS-PAGE, where denser Endo BI-1 bands in the supernatants were
present in epoxy and adsorption methods, while amino-based immobilization had no visible band
(Figure S1). Differences between immobilization yields for covalent and adsorption immobilization
methods have been observed previously and can possibly be attributed to the strength of interaction
between hydrophobic interactions and covalent bonds [18]. However, low binding using epoxide-
activated resin may be due to non-optimal methods to identify the ideal pH, time, and temperature
for increased binding [19]. Further investigation into those conditions may improve protein
immobilization yield for the epoxide-activated resin. Figure 1. Protein immobilization yield for amino, adsorption, and epoxy immobilization methods,
measured fluorometrically by Qubit Protein Assay Kit. Error bars represent one standard deviation
and means followed by different letters (a, b, and c) are statistically different at p < 0.05. Figure 1. Protein immobilization yield for amino, adsorption, and epoxy immobilization methods,
measured fluorometrically by Qubit Protein Assay Kit. 1. Introduction In addition to the
development of an immobilization method based on protein immobilization yield and enzyme activity,
the effects of the immobilization method on pH and temperature sensitivity of the immobilized enzyme 3 of 15
nzyme
of the Catalysts 2018, 8, 278
to the developmen
activity, the effec were evaluated. Enzyme reusability was also evaluated. Significant efforts were directed towards
the development of sensitive and high-throughput analytical methods: in particular, MALDI-ToF
MS was used to study the rate and specificity of the deglycosylation reaction kinetics on RNase B,
and nano-LC-Chip Q-ToF MS was used to characterize the diverse pool of N-glycans released from
bovine colostrum whey proteins using free and immobilized enzyme. bovine colostrum whey proteins. Significant efforts were directed towards the development of
sensitive and high-throughput analytical methods: in particular, MALDI-ToF MS was used to study
the rate and specificity of the deglycosylation reaction kinetics on RNase B, and nano-LC-Chip Q-ToF
MS was used to characterize the diverse pool of N-glycans released from bovine colostrum whey
proteins using free and immobilized enzyme. 2.1. Protein Immobilization Yield
2.1. Protein Immobilization Yield Error bars represent one standard deviation
and means followed by different letters (a, b, and c) are statistically different at p < 0.05. Figure 1. Protein immobilization yield for amino, adsorption, and epoxy immobilization methods,
measured fluorometrically by Qubit Protein Assay Kit. Error bars represent one standard deviation
and means followed by different letters (a, b, and c) are statistically different at p < 0.05. Figure 1. Protein immobilization yield for amino, adsorption, and epoxy immobilization methods,
measured fluorometrically by Qubit Protein Assay Kit. Error bars represent one standard deviation
and means followed by different letters (a, b, and c) are statistically different at p < 0.05. Figure 1. Protein immobilization yield for amino, adsorption, and epoxy immobilization methods,
measured fluorometrically by Qubit Protein Assay Kit. Error bars represent one standard deviation
and means followed by different letters (a, b, and c) are statistically different at p < 0.05. Figure 1. Protein immobilization yield for amino, adsorption, and epoxy immobilization methods,
measured fluorometrically by Qubit Protein Assay Kit. Error bars represent one standard deviation
and means followed by different letters (a, b, and c) are statistically different at p < 0.05. 2.2. Relative Quantification of N-glycans by MALDI-ToF MS
2.2. Relative Quantification of N-glycans by MALDI-ToF MS An additional goal of this research was to identify a sensitive, specific, and rapid method to
determine relative quantities of N-glycans to facilitate assessment of enzyme activity. Spectrophotometric methods involving phenol-sulfuric acid have been used, but lack selectivity for
certain saccharide types and tend to be affected by other compounds present in the sample, not only
An additional goal of this research was to identify a sensitive, specific, and rapid method to determine
relative quantities of N-glycans to facilitate assessment of enzyme activity. Spectrophotometric methods
involving phenol-sulfuric acid have been used, but lack selectivity for certain saccharide types and
tend to be affected by other compounds present in the sample, not only the N-glycan products of this
reaction [20–22]. Additionally, the diverse array of monosaccharides present in N-glycans adds a layer 4 of 15
harides
assays Catalysts 2018, 8, 278
the N-glycan pro
t i
N
l of complexity leading to inconsistent responses in these assays [21]. As an alternative, matrix-assisted
laser desorption/ionization time-of-flight mass spectrometry MALDI-ToF MS has been used for relative
quantification of various carbohydrates [23,24]. It is known that MALDI-ToF, albeit rapid and effective in
providing results, may suffer from low reproducibility due to inconsistent crystallization of the samples
on the target plate, leading to shot-to-shot variation. These disadvantages can be overcome by using an
internal standard [25]. Additionally, MALDI has been used to study enzymatic reactions including both
large and small substrates/products [26–28]. [21]. As an alternative, matrix assisted laser desorption/ionization time of flight mass spectrometry
MALDI-ToF MS has been used for relative quantification of various carbohydrates [23,24]. It is
known that MALDI-ToF, albeit rapid and effective in providing results, may suffer from low
reproducibility due to inconsistent crystallization of the samples on the target plate, leading to shot-
to-shot variation. These disadvantages can be overcome by using an internal standard [25]. Additionally, MALDI has been used to study enzymatic reactions including both large and small
substrates/products [26–28]. Linearity of the 3-FL (3’-fucosyllactose)-spiked released N-glycan system was evaluated. An g
p
Linearity of the 3-FL (3’-fucosyllactose)-spiked released N-glycan system was evaluated. An example
of a typical annotated mass spectrum of N-glycans can be seen in Figure 2. The theoretical m/z for sodiated
3-FL, Man 5, Man 6, Man 7, and Man 8 are 511.16, 1054.34, 1216.40, 1378.45, and 1540.50 respectively,
accounting for one fewer N-acetylglucosamine residue according to cleavage by Endo BI-1. 2.2. Relative Quantification of N-glycans by MALDI-ToF MS
2.2. Relative Quantification of N-glycans by MALDI-ToF MS The most
abundant glycan released from RNase B was Man5 (m/z 1054), with larger glycans being less abundant,
which was consistent with previous literature [29]. Bovine RNase B was chosen as a model glycoprotein
due to its single N-glycosylation site and straightforward glycan composition. Linearity of the 3 FL (3 fucosyllactose) spiked released N glycan system was evaluated. An
example of a typical annotated mass spectrum of N-glycans can be seen in Figure 2. The theoretical
m/z for sodiated 3-FL, Man 5, Man 6, Man 7, and Man 8 are 511.16, 1054.34, 1216.40, 1378.45, and
1540.50 respectively, accounting for one fewer N-acetylglucosamine residue according to cleavage by
Endo BI-1. The most abundant glycan released from RNase B was Man5 (m/z 1054), with larger
glycans being less abundant, which was consistent with previous literature [29]. Bovine RNase B was
chosen as a model glycoprotein due to its single N-glycosylation site and straightforward glycan
composition. Figure 2. MALDI-ToF mass spectrum for 3-FL (m/z 511)-spiked N-glycans released from RNase B
using Endo BI-1. Figure 2. MALDI-ToF mass spectrum for 3-FL (m/z 511)-spiked N-glycans released from RNase B using
Endo BI-1. Figure 2. MALDI-ToF mass spectrum for 3-FL (m/z 511)-spiked N-glycans released from RNase B
using Endo BI-1. Figure 2. MALDI-ToF mass spectrum for 3-FL (m/z 511)-spiked N-glycans released from RNase B using
Endo BI-1. Over the examined order of magnitude range, the normalized response plotted against N-
glycan/I.S. (internal standard) ratio was linear (R2 = 0.995, Figure S2). A high R2 value indicates that
analysis of released N-glycans in presence of I.S. analyzed by MALDI-ToF MS is a suitable tool for
rapid relative quantification of N-glycans needed for evaluating kinetic parameters over a substantial
range of concentrations and amounts of reactant. Over the examined order of magnitude range, the normalized response plotted against N-glycan/I.S. (internal standard) ratio was linear (R2 = 0.995, Figure S2). A high R2 value indicates that analysis of released
N-glycans in presence of I.S. analyzed by MALDI-ToF MS is a suitable tool for rapid relative quantification
of N-glycans needed for evaluating kinetic parameters over a substantial range of concentrations and
amounts of reactant. 2.4. Temperature and pH Sensitivity of Immobilized Enzyme Using RNase B
2.4. Temperature and pH Sensitivity of Immobilized Enzyme Using RNase B Enzyme properties can change with respect to pH and temperature sensitivity when
immobilized [31,32]. Temperature sensitivity was evaluated at pH 5 with 20 mM Na2HPO4 buffer at
temperatures from 45 to 85 °C (Figure 4a). The optimal temperature for both free and immobilized
Endo BI-1 was 65 °C. These results differ from what was previously reported by Karav et al. [33], in
which a lower optimum temperature of 52 °C was identified for the free enzyme. This discrepancy
could be attributed to different reaction conditions used in both studies. While RNase B and a short
reaction time (20 min) were used in the present work, bovine colostrum whey protein and longer
reaction times (15–475 min) were evaluated in the previous study. At the optimum temperature
identified (65 °C), the immobilized enzyme retained 63% of the activity compared with the free
enzyme. In each experimental condition of both temperature and pH, the activity of immobilized
Endo BI-1 was lower than that of the free enzyme, at all conditions. For both free and immobilized
enzymes, activities decreased at temperatures above 65 °C. T
l
H
i i i
20
M N
HPO b ff
dj
d
H 3 5 7
d 9 R
i
Enzyme properties can change with respect to pH and temperature sensitivity when
immobilized [31,32]. Temperature sensitivity was evaluated at pH 5 with 20 mM Na2HPO4 buffer at
temperatures from 45 to 85 ◦C (Figure 4a). The optimal temperature for both free and immobilized
Endo BI-1 was 65 ◦C. These results differ from what was previously reported by Karav et al. [33],
in which a lower optimum temperature of 52 ◦C was identified for the free enzyme. This discrepancy
could be attributed to different reaction conditions used in both studies. While RNase B and a short
reaction time (20 min) were used in the present work, bovine colostrum whey protein and longer
reaction times (15–475 min) were evaluated in the previous study. At the optimum temperature
identified (65 ◦C), the immobilized enzyme retained 63% of the activity compared with the free
enzyme. In each experimental condition of both temperature and pH, the activity of immobilized
Endo BI-1 was lower than that of the free enzyme, at all conditions. For both free and immobilized
enzymes, activities decreased at temperatures above 65 ◦C. To evaluate pH sensitivity, 20 mM Na2HPO4 buffer was adjusted to pH 3, 5, 7, and 9. 2.4. Temperature and pH Sensitivity of Immobilized Enzyme Using RNase B
2.4. Temperature and pH Sensitivity of Immobilized Enzyme Using RNase B Reactions
with free and immobilized Endo BI-1 and RNase B were carried out at 45 °C for 20 min, and the
normalized activity can be seen in Figure 4b. The optimal pH for free Endo BI-1 was 5, in agreement
with previous reports [9], while for the immobilized enzyme a shift to a neutral pH (7.0) was
observed. At the optimal pH for the free (pH 5) and immobilized enzyme (pH 7), the immobilized
enzyme retained approximately 62% of the activity of the free enzyme at 20 min of reaction. However,
the immobilized enzyme was more resilient in the range of pH 7 to 9, where reduced differences in
the activities of the free and immobilized enzyme were observed. Indeed, at pH 9, the activities of
free and immobilized Endo BI-1 were not statistically different (Figure 4a). Our results are in
agreement with several reports in the literature where a shift on the working pH of the immobilized
enzyme to a neutral or more alkaline pH was observed, with the same being attributed to changes in
the amine group during the covalent binding [31,34]. In the context of scaling up N-glycan release
using bovine milk proteins the optimum pH of the immobilized enzyme (pH 7) presents a clear
y
p
To evaluate pH sensitivity, 20 mM Na2HPO4 buffer was adjusted to pH 3, 5, 7, and 9. Reactions
with free and immobilized Endo BI-1 and RNase B were carried out at 45 ◦C for 20 min, and the
normalized activity can be seen in Figure 4b. The optimal pH for free Endo BI-1 was 5, in agreement
with previous reports [9], while for the immobilized enzyme a shift to a neutral pH (7.0) was observed. At the optimal pH for the free (pH 5) and immobilized enzyme (pH 7), the immobilized enzyme
retained approximately 62% of the activity of the free enzyme at 20 min of reaction. However,
the immobilized enzyme was more resilient in the range of pH 7 to 9, where reduced differences
in the activities of the free and immobilized enzyme were observed. Indeed, at pH 9, the activities of
free and immobilized Endo BI-1 were not statistically different (Figure 4a). 2.3. Comparing Immobilized Enzyme Activities
2.3. Comparing Immobilized Enzyme Activities The activity of the enzyme immobilized by each method (amino, epoxy, and adsorption) was
evaluated on RNase B, and reacted for 90 min at 45 °C and pH 5.0. The normalized relative abundance
of released N-glycans was determined by MALDI-ToF MS, with activity based on the amount of
glycans released by the free enzyme (Figure 3). Free enzyme had the highest activity, with amino,
epoxy, and adsorption methods retaining 73, 51, and 57% activity, respectively. Diminished activity
upon immobilization is common, and has been reported previously with glutaraldehyde-mediated
The activity of the enzyme immobilized by each method (amino, epoxy, and adsorption) was
evaluated on RNase B, and reacted for 90 min at 45 ◦C and pH 5.0. The normalized relative abundance
of released N-glycans was determined by MALDI-ToF MS, with activity based on the amount of
glycans released by the free enzyme (Figure 3). Free enzyme had the highest activity, with amino, epoxy,
and adsorption methods retaining 73, 51, and 57% activity, respectively. Diminished activity upon
immobilization is common, and has been reported previously with glutaraldehyde-mediated binding
and hydrophobic adsorption [30]. A combination of diffusional/steric hindrances and possible binding 5 of 15
5 of 15 Catalysts 2018, 8, 278
Catalysts 2018, 8, x FO and occlusion of enzyme active sites may explain the reduction in enzyme activity [16]. Additionally,
heterogeneous catalysis impacts enzyme activity. The amino-based immobilization was selected for
all subsequent experiments due to its higher activity compared with epoxy and adsorption methods,
as well as its improved catalyst density on the resin. g
y
p
p
possible binding and occlusion of enzyme active sites may explain the reduction in enzyme activity
[16]. Additionally, heterogeneous catalysis impacts enzyme activity. The amino-based
immobilization was selected for all subsequent experiments due to its higher activity compared with
epoxy and adsorption methods, as well as its improved catalyst density on the resin. Figure 3. Relative activity of free or immobilized Endo BI-1 (on amino, epoxy, and adsorption resins)
on the glycoprotein RNase B. Error bars represent one standard deviation. Means followed by
different letters (a, b, and c) are statistically different at p < 0.05. Figure 3. Relative activity of free or immobilized Endo BI-1 (on amino, epoxy, and adsorption resins)
on the glycoprotein RNase B. Error bars represent one standard deviation. Means followed by different
letters (a, b, and c) are statistically different at p < 0.05. Figure 3. 2.3. Comparing Immobilized Enzyme Activities
2.3. Comparing Immobilized Enzyme Activities Relative activity of free or immobilized Endo BI-1 (on amino, epoxy, and adsorption resins)
on the glycoprotein RNase B. Error bars represent one standard deviation. Means followed by
different letters (a, b, and c) are statistically different at p < 0.05. Figure 3. Relative activity of free or immobilized Endo BI-1 (on amino, epoxy, and adsorption resins)
on the glycoprotein RNase B. Error bars represent one standard deviation. Means followed by different
letters (a, b, and c) are statistically different at p < 0.05. 2.5. Enzyme Kinetics Using RNase B
To ai
o e ua titati e i
i
2.5. Enzyme Kinetics Using RNase B To gain more quantitative insight into the mechanistic changes of the enzymatic deglycosylation
upon immobilization, the kinetic parameters (Vmax, Km) were estimated using linearized plotting
techniques (Lineweaver-Burk and Hanes Woolf) to reduce the error of non-linear estimation (Figure
5). Vmax, the maximum forward velocity of the reaction, decreased from 0.551 glycan/min to 0.076
glycan/min (Lineweaver Burk estimation) upon immobilization of Endo BI-1 as calculated on RNase
B (Figure 6). The Michaelis constant (Km, a measure of substrate affinity) also decreased upon
immobilization from 2.27 mg/mL for the free enzyme to 0.299 mg/mL once Endo BI-1 was
immobilized on the amino-activated methacrylate resin. The goodness-of-fit for the Lineweaver Burk
linear model (R2) for free and immobilized enzyme was 0.89 and 0.96 respectively. Hanes Woolf linear
estimation revealed similar trends (0.333 and 0.081 Vmax values, 1.17 and 0.345 Km values for free and
immobilized enzymes, respectively) to Lineweaver Burk. Decreased Vmax is typical of enzyme
immobilization [32,36], wherein lower maximum velocity is likely due to limited access of the large,
macromolecular substrate to the active site of the immobilized enzyme. The immobilized enzyme is
on a molecular tether, and steric hindrances limit access to more occluded glycan sites on the
substrate with respect to the three-dimensional structure. As a point of comparison, β-1,4-
galactosidase, with a molecular weight of approximately 465 kDa in its tetrameric form, with lactose
as substrate at 360 Da has an overall substrate/enzyme size ratio of 0.0008:1, whereas for RNase B and
Endo BI-1, the substrate/enzyme size ratio is approximately 0.32:1. This suggests a more limited
diffusion and increased steric challenge with respect to active site availability. Increased apparent affinity accompanied by decreased Vmax is consistent with uncompetitive
To gain more quantitative insight into the mechanistic changes of the enzymatic deglycosylation upon
immobilization, the kinetic parameters (Vmax, Km) were estimated using linearized plotting techniques
(Lineweaver-Burk and Hanes Woolf) to reduce the error of non-linear estimation (Figure 5). Vmax,
the maximum forward velocity of the reaction, decreased from 0.551 glycan/min to 0.076 glycan/min
(Lineweaver Burk estimation) upon immobilization of Endo BI-1 as calculated on RNase B (Figure 6). The Michaelis constant (Km, a measure of substrate affinity) also decreased upon immobilization from
2.27 mg/mL for the free enzyme to 0.299 mg/mL once Endo BI-1 was immobilized on the amino-activated
methacrylate resin. 2.4. Temperature and pH Sensitivity of Immobilized Enzyme Using RNase B
2.4. Temperature and pH Sensitivity of Immobilized Enzyme Using RNase B Our results are in agreement
with several reports in the literature where a shift on the working pH of the immobilized enzyme to a
neutral or more alkaline pH was observed, with the same being attributed to changes in the amine
group during the covalent binding [31,34]. In the context of scaling up N-glycan release using bovine 6 of 15 Catalysts 2018, 8, 278 milk proteins, the optimum pH of the immobilized enzyme (pH 7), presents a clear advantage over the
free enzyme, considering that milk and dairy streams naturally have a pH close to neutral and would
not necessitate the use of buffers for pH adjustment, which can become problematic and costly at large
scale. Literature suggests that immobilization alters temperature and pH sensitivity by stabilizing the
enzyme conformation and creating stable microenvironments around the enzyme or close to its active
site [35]. The data here presented provides valuable information for future scale-up of the process and
testing other substrates for glycan release. Catalysts 2018, 8, x FOR PEER REVIEW
6 of 15
advantage over the free enzyme, considering that milk and dairy streams naturally have a pH close
to neutral and would not necessitate the use of buffers for pH adjustment, which can become
problematic and costly at large scale. Literature suggests that immobilization alters temperature and
pH sensitivity by stabilizing the enzyme conformation and creating stable microenvironments
around the enzyme or close to its active site [35]. The data here presented provides valuable
information for future scale-up of the process and testing other substrates for glycan release. Figure 4. Mean relative activity of free and immobilized enzyme at different (a) temperature and (b)
pH values using RNase B. Error bars represent one standard deviation and means, within the same
chart, followed by different letters (a, b, c, d, e, and f) are statistically different at p < 0.05. Figure 4. Mean relative activity of free and immobilized enzyme at different (a) temperature and
(b) pH values using RNase B. Error bars represent one standard deviation and means, within the same
chart, followed by different letters (a, b, c, d, e, and f) are statistically different at p < 0.05. Figure 4. Mean relative activity of free and immobilized enzyme at different (a) temperature and (b)
pH values using RNase B. 2.4. Temperature and pH Sensitivity of Immobilized Enzyme Using RNase B
2.4. Temperature and pH Sensitivity of Immobilized Enzyme Using RNase B Error bars represent one standard deviation and means, within the same
chart, followed by different letters (a, b, c, d, e, and f) are statistically different at p < 0.05. Figure 4. Mean relative activity of free and immobilized enzyme at different (a) temperature and
(b) pH values using RNase B. Error bars represent one standard deviation and means, within the same
chart, followed by different letters (a, b, c, d, e, and f) are statistically different at p < 0.05. 2.6. Immobilized Enzyme Reusability with RNase B
A
b
i l
d
f
i
bil
2.6. Immobilized Enzyme Reusability with RNase B A substantial advantage of enzyme immobilization is the potential reusability of the catalyst. Amino-mediated immobilized Endo BI-1 was subjected to ten 20-min reuse cycles at 45 and 65 °C to
determine the effects of different temperatures on the activity and reusability of the immobilized
enzyme (Figure 6). Enzyme activity was higher at 65 °C compared with 45 °C. At 45 °C, the
mmobilized enzyme activity corresponded to approximately 65% of the enzyme activity at 65 °C. Despite higher enzyme activity at 65 °C, a higher decrease in the enzyme activity was observed after
each reuse cycle. At 65 °C, enzyme activity was reduced to 63% of its original value after one cycle of
reuse, and by the sixth use cycle, it had been reduced to and stabilized at 10 to 15% of its original
activity. Conversely, at 45 °C, the enzyme activity remained consistent up to three reuse cycles. Despite the initial higher enzyme activity at 65 °C, from the third cycle onward reactions at 45 °C had
greater residual enzyme activity than reactions performed at 65 °C. These results indicate that
although 65 °C is optimal for initial glycan release, it does not favor repeated enzyme use for longer
reaction times. It is possible to reuse the immobilized enzyme up to seven cycles at 45 °C with
significant retention of its activity (50% from the initial activity). Our results are in agreement with
several reports in the literature, where the use of lower temperatures (e.g., 37 °C) has favored high
reusability, even though higher temperatures may favor more rapid catalysis [38,39]. Regarding an
ndustrially-relevant enzyme with macromolecular substrates Chauhan reported a loss of over 85%
A substantial advantage of enzyme immobilization is the potential reusability of the catalyst. Amino-mediated immobilized Endo BI-1 was subjected to ten 20-min reuse cycles at 45 and 65 ◦C to
determine the effects of different temperatures on the activity and reusability of the immobilized enzyme
(Figure 6). Enzyme activity was higher at 65 ◦C compared with 45 ◦C. At 45 ◦C, the immobilized enzyme
activity corresponded to approximately 65% of the enzyme activity at 65 ◦C. Despite higher enzyme
activity at 65 ◦C, a higher decrease in the enzyme activity was observed after each reuse cycle. 2.5. Enzyme Kinetics Using RNase B
To ai
o e ua titati e i
i
2.5. Enzyme Kinetics Using RNase B The goodness-of-fit for the Lineweaver Burk linear model (R2) for free and immobilized
enzyme was 0.89 and 0.96 respectively. Hanes Woolf linear estimation revealed similar trends (0.333 and
0.081 Vmax values, 1.17 and 0.345 Km values for free and immobilized enzymes, respectively) to Lineweaver
Burk. Decreased Vmax is typical of enzyme immobilization [32,36], wherein lower maximum velocity is
likely due to limited access of the large, macromolecular substrate to the active site of the immobilized
enzyme. The immobilized enzyme is on a molecular tether, and steric hindrances limit access to more
occluded glycan sites on the substrate with respect to the three-dimensional structure. As a point of
comparison, β-1,4-galactosidase, with a molecular weight of approximately 465 kDa in its tetrameric form,
with lactose as substrate at 360 Da has an overall substrate/enzyme size ratio of 0.0008:1, whereas for
RNase B and Endo BI-1, the substrate/enzyme size ratio is approximately 0.32:1. This suggests a more
limited diffusion and increased steric challenge with respect to active site availability. Catalysts 2018, 8, 278 7 of 15 Catalysts 2018, 8, 278 Increased apparent affinity accompanied by decreased Vmax is consistent with uncompetitive
inhibition. Possible explanations of this phenomenon are the formation of a more stable enzyme-substrate
complex between Endo BI-1 and RNase B upon immobilization, or simply deglycosylated RNase B
may have difficulty diffusing away from immobilized Endo BI-1 upon interaction with the active site. Both stable enzyme-substrate complexes and diffusional challenges would increase apparent affinity and
inhibit forward reaction progress and are documented phenomena [36,37]. Catalysts 2018, 8, x FOR PEER REVIEW
7 of 15
RNase B may have difficulty diffusing away from immobilized Endo BI-1 upon interaction with the
active site. Both stable enzyme-substrate complexes and diffusional challenges would increase
apparent affinity and inhibit forward reaction progress and are documented phenomena [36,37]. Enzymes with diverse, macromolecular substrates may exhibit substrate-dependent kinetic
behavior. For example, when Endo BI-1 was tested on different glycoproteins, Vmax and Km also
varied. The Michaelis constants for RNase B, lactoferrin, and concentrated whey proteins were 0.25,
0.43, and 0.90 mg/mL respectively, potentially due to different sizes, purities, and physicochemical
properties of the glycoproteins [20]. The overall increase in Km for Endo BI-1 in this study (0.33 vs. 025 mg/mL) is likely related to differences in reaction conditions and analytical methods used in both
studies. 2.5. Enzyme Kinetics Using RNase B
To ai
o e ua titati e i
i
2.5. Enzyme Kinetics Using RNase B While this study utilized MALDI-ToF MS to measure the hydrolytic efficiency of Endo BI-1,
previous studies utilized a spectrophotometric phenol-sulfuric assay, wherein the diverse array of
monosaccharides present in N-glycans from lactoferrin and other whey proteins could have yielded
different responses. apparent affinity and inhibit forward reaction progress and are documented phenomena [36,37]. Enzymes with diverse, macromolecular substrates may exhibit substrate-dependent kinetic
behavior. For example, when Endo BI-1 was tested on different glycoproteins, Vmax and Km also
varied. The Michaelis constants for RNase B, lactoferrin, and concentrated whey proteins were 0.25,
0.43, and 0.90 mg/mL respectively, potentially due to different sizes, purities, and physicochemical
properties of the glycoproteins [20]. The overall increase in Km for Endo BI-1 in this study (0.33 vs. 025 mg/mL) is likely related to differences in reaction conditions and analytical methods used in both
studies. While this study utilized MALDI-ToF MS to measure the hydrolytic efficiency of Endo BI-1,
previous studies utilized a spectrophotometric phenol-sulfuric assay, wherein the diverse array of
monosaccharides present in N-glycans from lactoferrin and other whey proteins could have yielded
different responses. (a)
(b)
Figure 5. Lineweaver Burk (1/[S] (mL/mg) vs. 1/v (mL × min/mg)) (a) and Hanes–Woolf ([S] (mg/mL)
vs. s/v (min)) (b) linearized plotting techniques for the estimation of kinetic parameters of free and
immobilized EndoBI-1 on RNase B. Figure 5. Lineweaver Burk (1/[S] (mL/mg) vs. 1/v (mL × min/mg)) (a) and Hanes–Woolf ([S]
(mg/mL) vs. s/v (min)) (b) linearized plotting techniques for the estimation of kinetic parameters of
free and immobilized EndoBI-1 on RNase B. (a) (b) (b) (a) Figure 5. Lineweaver Burk (1/[S] (mL/mg) vs. 1/v (mL × min/mg)) (a) and Hanes–Woolf ([S] (mg/mL)
vs. s/v (min)) (b) linearized plotting techniques for the estimation of kinetic parameters of free and
mmobilized EndoBI-1 on RNase B. Figure 5. Lineweaver Burk (1/[S] (mL/mg) vs. 1/v (mL × min/mg)) (a) and Hanes–Woolf ([S]
(mg/mL) vs. s/v (min)) (b) linearized plotting techniques for the estimation of kinetic parameters of
free and immobilized EndoBI-1 on RNase B. 2.7. Release of N-glycans from Bovine whey Proteins
2.7. Release of N-glycans from Bovine whey Proteins 2.7. Release of N-glycans from Bovine whey Proteins
2.7. Release of N-glycans from Bovine whey Proteins Cheese whey, a co-product of the dairy industry, is a source of N-linked glycoproteins including
lactoferrin, lactoperoxidase, transferrin, and various immunoglobulins [12]. Free and immobilized
Endo BI-1 was incubated with whey proteins purified from bovine colostrum at pH 7 at 45 °C for 1
h. The glycans were purified and characterized by nano LC-Chip Q-ToF mass spectrometry and each
compound’s relative abundance was determined and further classified based on the class of glycan
to which each compound belongs (Figure 7). O e all t
e ty i u i ue N
ly a
t u tu e
e e ide tified a d o fi
ed by ta de
a
Cheese whey, a co-product of the dairy industry, is a source of N-linked glycoproteins including
lactoferrin, lactoperoxidase, transferrin, and various immunoglobulins [12]. Free and immobilized
Endo BI-1 was incubated with whey proteins purified from bovine colostrum at pH 7 at 45 ◦C for 1 h. The glycans were purified and characterized by nano LC-Chip Q-ToF mass spectrometry and each
compound’s relative abundance was determined and further classified based on the class of glycan to
which each compound belongs (Figure 7). Overall, twenty-six unique N-glycan structures were identified and confirmed by tandem mass
spectra. The N-glycans were characterized based on their monosaccharide composition, which
included neutral (containing only hexose and N-acetylhexosamine residues), sialylated, fucosylated,
and fucosylated and sialylated. For neutral glycans the immobilized enzyme released eight times
higher relative amounts than free Endo BI-1. For the class fucosylated and sialylated, the free enzyme
released significantly higher amount of glycans (approximately double) compared with the free
enzyme. Free enzyme released 50% more of each fucosylated and sialylated glycan class. For glycans
with both sialylation and fucosylation, the free enzyme released nearly double the relative amount
of glycans compared with immobilization. It is worth mentioning that fucosylated and sialylated
glycans were present in lowest amounts. Both sialylated and neutral glycans represented the most
abundant glycan types released from colostrum whey proteins. Importantly, unique glycan
compositions should only be compared between samples, and not among the various N-glycan
structures themselves. In fact, differences in ionization efficiency due to diverse monosaccharide
composition hinder direct comparisons in the absence of identical pure standards. 2.6. Immobilized Enzyme Reusability with RNase B
A
b
i l
d
f
i
bil
2.6. Immobilized Enzyme Reusability with RNase B At 65 ◦C,
enzyme activity was reduced to 63% of its original value after one cycle of reuse, and by the sixth use
cycle, it had been reduced to and stabilized at 10 to 15% of its original activity. Conversely, at 45 ◦C,
the enzyme activity remained consistent up to three reuse cycles. Despite the initial higher enzyme activity
at 65 ◦C, from the third cycle onward reactions at 45 ◦C had greater residual enzyme activity than reactions
performed at 65 ◦C. These results indicate that although 65 ◦C is optimal for initial glycan release, it does
not favor repeated enzyme use for longer reaction times. It is possible to reuse the immobilized enzyme up
to seven cycles at 45 ◦C with significant retention of its activity (50% from the initial activity). Our results
are in agreement with several reports in the literature, where the use of lower temperatures (e.g., 37 ◦C)
has favored high reusability, even though higher temperatures may favor more rapid catalysis [38,39]. Regarding an industrially-relevant enzyme with macromolecular substrates, Chauhan reported a loss 8 of 15 Catalysts 2018, 8, 278 of over 85% of polygalacturonase activity after 6 cycles using adsorption onto celite [40]. Additionally,
reaction time and temperature affect the selective release of N-glycans from more complex and diverse
glycoproteins, which may determine the choice of the reaction temperature thus affecting reusability
of the immobilized enzyme [41]. Importantly, fouling with a protein-based substrate may also impact
reusability [42]. Catalysts 2018, 8, x FOR PEER REVIEW
8 of 15 Figure 6. Mean relative activity of immobilized Endo BI-1 on RNase B following several reuse cycles. Error bars represent one standard deviation. Figure 6. Mean relative activity of immobilized Endo BI-1 on RNase B following several reuse cycles. Error bars represent one standard deviation. Figure 6. Mean relative activity of immobilized Endo BI-1 on RNase B following several reuse cycles. Error bars represent one standard deviation. Figure 6. Mean relative activity of immobilized Endo BI-1 on RNase B following several reuse cycles. Error bars represent one standard deviation. 2.7. Release of N-glycans from Bovine whey Proteins
2.7. Release of N-glycans from Bovine whey Proteins These large differences are the main drivers of the substantial differences in
relative abundance of neutral glycans. Both 3_3_0_0_0 and 4_3_0_1_0 were identified as the most
abundant glycans released from a similar protein source using Endo BI-1 [41]. Although high mannose
compounds like 5_1_0_0_0 and 6_1_0_0_0 were previously found in the bovine and human milk
N-glycome, likely deriving from bovine lactoferrin, they were not previously described as being
released from bovine colostrum whey proteins with free Endo BI-1 [41,43]. Catalysts 2018, 8, x FOR PEER REVIEW
9 of 15
Despite the many technological breakthroughs deglycosylation is still not a well characterized 3_3_0_0_0 and 5_1_0_0_0 for immobilized enzyme compared with free enzyme were 27 and 14 times
higher, respectively. These large differences are the main drivers of the substantial differences in
relative abundance of neutral glycans. Both 3_3_0_0_0 and 4_3_0_1_0 were identified as the most
abundant glycans released from a similar protein source using Endo BI-1 [41]. Although high mannose
compounds like 5_1_0_0_0 and 6_1_0_0_0 were previously found in the bovine and human milk
N-glycome, likely deriving from bovine lactoferrin, they were not previously described as being
released from bovine colostrum whey proteins with free Endo BI-1 [41,43]. Catalysts 2018, 8, x FOR PEER REVIEW
9 of 15
Despite the many technological breakthroughs deglycosylation is still not a well characterized y p
Despite the many technological breakthroughs, deglycosylation is still not a well-characterized
process. Recently, the kinetics of the commercial enzyme peptide-N-glycosidase F (PNGase F) were
investigated to understand the impact of sialylation on the release of N-glycans [44]. Presence of sialic
acid slowed glycan release, while fucosylation also affected the deglycosylation rate. Because RNase B
contains a single glycosylation site and high mannose type glycans, these differences would not be
observed. However, when using a different enzyme (Endo BI-1) and complex substrates like bovine
colostrum whey proteins under different conditions, kinetics and deglycosylation behavior would
likely be substantially different. Additionally, the presence of methacrylate resin with pore diameters
within the range of 300 to 600 Å may facilitate binding of small, neutral glycans such as 3_3_0_0_0
and 5_1_0_0_0 or the protein regions containing them, favoring their release compared with free Endo
BI-1. Antennary moieties such as fucose or sialic acid could also change physicochemical interactions
between the glycan and the methacrylate supports as well as the glycan with the immobilized enzyme
itself. 2.7. Release of N-glycans from Bovine whey Proteins
2.7. Release of N-glycans from Bovine whey Proteins The ability to modulate glycan release based on reaction conditions has been previously described
by Le Parc et al. [41]. Additional changes on the glycan release resulting from the immobilization
method used suggests a high degree of complexity concerning enzymatic deglycosylation. Despite the many technological breakthroughs, deglycosylation is still not a well-characterized
process. Recently, the kinetics of the commercial enzyme peptide-N-glycosidase F (PNGase F) were
investigated to understand the impact of sialylation on the release of N-glycans [44]. Presence of sialic
acid slowed glycan release, while fucosylation also affected the deglycosylation rate. Because RNase
B contains a single glycosylation site and high mannose type glycans, these differences would not be
observed. However, when using a different enzyme (Endo BI-1) and complex substrates like bovine
colostrum whey proteins under different conditions, kinetics and deglycosylation behavior would
likely be substantially different. Additionally, the presence of methacrylate resin with pore diameters
within the range of 300 to 600 Å may facilitate binding of small, neutral glycans such as 3_3_0_0_0
and 5_1_0_0_0 or the protein regions containing them, favoring their release compared with free
Endo BI-1. Antennary moieties such as fucose or sialic acid could also change physicochemical
interactions between the glycan and the methacrylate supports as well as the glycan with the
immobilized enzyme itself. The ability to modulate glycan release based on reaction conditions has
been previously described by Le Parc et al. [41]. Additional changes on the glycan release resulting
from the immobilization method used suggests a high degree of complexity concerning enzymatic
deglycosylation. Figure 7. Mean relative quantities of various classes of released N-glycans from bovine colostrum
whey proteins determined by nano-LC Chip Q-ToF MS/MS by free and immobilized Endo BI-1. Error
bars represent one standard deviation. * Represents significant differences within the same class of N-
glycans at p < 0.05. 3 Materials and Methods
Figure 7. Mean relative quantities of various classes of released N-glycans from bovine colostrum
whey proteins determined by nano-LC Chip Q-ToF MS/MS by free and immobilized Endo BI-1. Error
bars represent one standard deviation. * Represents significant differences within the same class of
N-glycans at p < 0.05. Figure 7. Mean relative quantities of various classes of released N-glycans from bovine colostrum
whey proteins determined by nano-LC Chip Q-ToF MS/MS by free and immobilized Endo BI-1. Error
bars represent one standard deviation. 2.7. Release of N-glycans from Bovine whey Proteins
2.7. Release of N-glycans from Bovine whey Proteins Th
N
l
t
t
l
d b
E d
BI 1
t b l t d
di
t
th
l
Overall, twenty-six unique N-glycan structures were identified and confirmed by tandem mass
spectra. The N-glycans were characterized based on their monosaccharide composition, which
included neutral (containing only hexose and N-acetylhexosamine residues), sialylated, fucosylated,
and fucosylated and sialylated. For neutral glycans the immobilized enzyme released eight times
higher relative amounts than free Endo BI-1. For the class fucosylated and sialylated, the free enzyme
released significantly higher amount of glycans (approximately double) compared with the free
enzyme. Free enzyme released 50% more of each fucosylated and sialylated glycan class. For glycans
with both sialylation and fucosylation, the free enzyme released nearly double the relative amount of
glycans compared with immobilization. It is worth mentioning that fucosylated and sialylated glycans
were present in lowest amounts. Both sialylated and neutral glycans represented the most abundant
glycan types released from colostrum whey proteins. Importantly, unique glycan compositions should
only be compared between samples, and not among the various N-glycan structures themselves. In fact, differences in ionization efficiency due to diverse monosaccharide composition hinder direct
comparisons in the absence of identical pure standards. The N-glycan structures released by Endo BI-1 were tabulated according to the glycan
composition notation Hex_HexNAc_Fuc_NeuAc_NeuGc, which correspond with the number of
hexose, N-acetylhexosamine, fucose, N-acetylneuraminic acid, and N-glycolylneuraminic acid
residues respectively (Table S1). The most abundant glycans released from colostrum whey proteins
by free Endo BI-1 were 4_3_0_1_0 and 3_3_0_1_0 while the most abundant glycan structures released
by immobilized Endo BI-1 were 3_3_0_0_0 and 5_1_0_0_0 (Man5). The relative abundance for
released 3 3 0 0 0 and 5 1 0 0 0 for immobilized enzyme compared with free enzyme were 27 and
The N-glycan structures released by Endo BI-1 were tabulated according to the glycan
composition notation Hex_HexNAc_Fuc_NeuAc_NeuGc, which correspond with the number of
hexose, N-acetylhexosamine, fucose, N-acetylneuraminic acid, and N-glycolylneuraminic acid residues
respectively (Table S1). The most abundant glycans released from colostrum whey proteins by free
Endo BI-1 were 4_3_0_1_0 and 3_3_0_1_0 while the most abundant glycan structures released by
immobilized Endo BI-1 were 3_3_0_0_0 and 5_1_0_0_0 (Man5). The relative abundance for released 9 of 15 Catalysts 2018, 8, 278 3_3_0_0_0 and 5_1_0_0_0 for immobilized enzyme compared with free enzyme were 27 and 14 times
higher, respectively. 3.3. Evaluating Protein Immobilization Yield Wash buffers from enzyme immobilization were treated with four volumes of −20 ◦C ethanol and
held at −30 ◦C for 60 min to precipitate proteins, after which the samples were centrifuged at 4000× g
for 30 min at 4 ◦C. The supernatant was discarded, and the pellet was resuspended in nanopure water. The total protein was loaded to the immobilization resin and the protein remaining in the wash buffer
was measured. The protein immobilization yield was determined by the following equation: Protein immobilization yield (%) = PL −Pw
PL
× 100% where PL is the mass of protein loaded and Pw is the mass of protein in the resin washing buffer
after immobilization [46]. Total protein was measured fluorometrically in triplicate using the Qubit
Protein Assay Kit (Life Technologies, Grand Island, NY, USA). Purified protein samples were also
analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) on 4–15% precast
polyacrylamide gels to visualize amount of protein not bound to immobilization resin [9]. 3.1. Enzyme Production Recombinant endo-β-(1,4)-N-acetylglucosaminidase (Endo BI-1) from Bifidobacterium longum
subsp. infantis ATCC 15697 from the University of California Davis Viticulture and Enology Culture
Collection (Davis, CA, USA) was cloned into E. coli using a pEcoTM-T7-cHis cloning kit (GeneTarget
Inc, San Diego, CA, USA). Endo BI-1 was expressed and isolated in a stock containing 65 mg/mL
enzymebased upon previous literature [33]. 3.2. Enzyme Immobilization Endo BI-1 was immobilized onto methacrylate resin according to manufacturer’s directions
(Purolite, Llantrisant, Wales, UK). Epoxide-activated (Lifetech ECR8204F), amino-activated (Lifetech
ECR8309F), and divinylbenzene (DVB, Lifetech ECR1030M) adsorption resins were used in this study. Covalent (epoxide- and amino-activated resins) and adsorption (divinylbenzene resin) immobilization
techniques were selected in this study due to limited macromolecular substrate diffusion through
entrapment matrices [45]. To activate the amino resin, 2% glutaraldehyde in 1X PBS as a crosslinking agent was added
to the resin prior to enzyme addition and incubated for 1 h, after which it was washed with 0.1X
phosphate-buffered saline (PBS) to remove excess glutaraldehyde. Endo BI-1 enzyme stock was added
to each activated resin (50 mg enzyme per gram activated resin, wet basis) in buffer mass equal to
four times the mass of the resin. For epoxide-activated immobilization, resin was incubated with
enzyme in 10X PBS buffer for 42 h. For adsorption-based immobilization, DVB resin was incubated
with enzyme for 18 h in 0.1X PBS buffer. Amino-based immobilization was incubated with enzyme in
0.1X PBS for 24 h, based on the manufacturers’ recommendations. Enzyme incubation took place at
25 ◦C and 100 rpm and each immobilization method was performed in triplicate. Following incubation,
resins were washed with four volumes of 0.1X PBS buffer. The wash buffer was collected and further
analyzed to determine the protein immobilization yield. 2.7. Release of N-glycans from Bovine whey Proteins
2.7. Release of N-glycans from Bovine whey Proteins * Represents significant differences within the same class of N-
glycans at p < 0.05. Figure 7. Mean relative quantities of various classes of released N-glycans from bovine colostrum
whey proteins determined by nano-LC Chip Q-ToF MS/MS by free and immobilized Endo BI-1. Error
bars represent one standard deviation. * Represents significant differences within the same class of
N-glycans at p < 0.05. 10 of 15 10 of 15 Catalysts 2018, 8, 278 3.7. Determination of Kinetic Parameters of Enzyme Using RNase B Relevant kinetic parameters relating to maximum reaction velocity (Vmax) and the Michaelis-
Menten constant (km) were determined by measuring the normalized glycan intensity at varying
substrate concentrations (0.1 to 1.1 g/L) in 20 mM Na2HPO4 buffer under optimal pH and temperature
conditions for the free and immobilized enzyme. Aliquots were taken in four-minute intervals up to
20 min. Linearized plotting (Lineweaver Burk and Hanes Woolf) were utilized to calculate Km and
Vmax [48,49]. 3.5. Preliminary Comparison of Immobilization Methods Efficiency Using Ribonuclease (RNase) B 3.5. Preliminary Comparison of Immobilization Methods Efficiency Using Ribonuclease (RNase) B Accounting for varying immobilization yields of the enzyme onto the beads, resin containing
40 µg immobilized Endo BI-1 or 40 µg free enzyme was reacted with 600 µg in 20 mM Na2HPO4 buffer
for 90 min at 45 ◦C at pH 5. The reaction was terminated with ethanol at −30 ◦C and glycans were
purified and characterized as described in the MALDI-ToF MS analysis section. Summed, normalized
MS intensities for the four most abundant glycans were normalized to 100% activity for the free
enzyme. One-way analysis of variance (ANOVA) was used along with Tukey’s multiple comparisons
test with GraphPad Prism 7 (Graphpad, La Jolla, CA, USA) to determine the statistical significance of
the data at p < 0.05. 3.6. Effect of Immobilization on Enzyme Resilience to pH and Thermal Changes To identify the optimum temperature for the immobilized enzyme, RNase B was incubated in
pH 5 buffer as described previously and free or immobilized enzyme was added [9]. The reaction
mixture (enzyme + substrate) was incubated for 20 min at temperatures ranging from 45 to 85 ◦C. Their respective deglycosylating activities were normalized and compared. The pH of 20 mM Na2HPO4 buffer was adjusted to 3, 5, 7, and 9 using 1 N HCl or NaOH. Free and
immobilized Endo BI-1 was reacted with RNase B at 45 ◦C at each pH for 20 min and the relative
activity, measured as the enzyme ability to deglycosylate RNase B, was measured by MALDI-ToF
MS. Experiments were conducted in triplicate and a two-way ANOVA was used along with Tukey’s
multiple comparisons in GraphPad Prism 7 to determine statistical significance of the data at p < 0.05. 3.4. N-glycan Analysis by MALDI-ToF Mass Spectrometry A microflex MALDI-ToF mass spectrometer (Bruker Daltonics GmbH, Bremen, Germany) was
used. Ten µL of purified N-glycan samples were spiked with five µL 100 mg/L of the human milk
oligosaccharide 3’-fucosyllactose (3-FL, Dextra Laboratories Inc., Reading, UK) as an internal standard. A one µL aliquot of spiked sample was combined with an equal volume of 2,5-dihydroxybenzoic acid
matrix (20 mg/mL in 30% acetonitrile, 0.1% trifluoroacetic acid) and 0.4 µL 1 mM NaCl was added 11 of 15 Catalysts 2018, 8, 278 to achieve ionization in positive mode. 0.5 µL spiked sample with matrix and NaCl was spotted in
duplicate onto a ground steel target plate and dried under vacuum. Mass calibration was conducted
using a polysaccharide ladder extracted from beer [47]. Ionization was carried out using a 337.1 nm laser, and detected in reflectron positive mode with
a reflector voltage of 20.02 kV in the mass range of 300 to 2500 m/z. The final spectra were the sum
of 2000 laser shots. MS intensities of four glycans (namely Man 5, Man 6, Man 7, Man 8) released
from bovine ribonuclease B (RNase B, Sigma Aldrich, St. Louis, MO, USA) were normalized to the
MS intensity of 3-FL internal standard, summed, and considered the normalized relative abundance
of glycan. g y
Linearity of the MALDI-ToF method was determined by preparing a stock of glycans released
from RNase B using free Endo BI-1. Following purification as described above, varying amounts
of released N-glycan was added to a constant amount of 3-FL internal standard and analyzed by
MALDI-ToF MS as described above. The summed, normalized intensity was plotted against the
volume of N-glycans added and a linear regression was fit to the data. 3.10. N-glycan Analysis by Mass Spectrometry Nano-LC-Chip Q ToF Samples were transferred to polypropylene vials and 6 µL were injected into an Agilent
6520 nano-LC-Chip quadrupole time-of-flight mass spectrometer (Q-ToF MS, Agilent Technologies,
Santa Clara, CA, USA). Glycans were further purified on a 9 mm × 75 µm PGC enrichment column and
then separated on a 43 mm × 75 µm PGC analytical column with binary solvents, gradient, and data
collection based on Karav et al. [7]. The following minor modifications were applied: scan range for
MS was 450 to 2500 m/z, and for MS/MS was 100 to 3200 m/z. Spectra in both MS and MS/MS modes
were collected at a rate of 0.63 spectra/s. The collision energy in the collision cell corresponded to a
slope of 1.8/100 Da. Chromatograms from the nano-LC-Chip Q ToF were curated and compounds identified in
Masshunter Qualitative Analysis software (version B.07.00 Agilent Technologies) using a previously
described bovine milk N-glycan bioinformatic library adjusted for one fewer N-acetylglucosamine in
the chitobiose core. The library assumed compositions including hexose (Hex), N-acetylhexosamine
(HexNAc), fucose (Fuc), N-acetylneuraminic acid (NeuAc), and N-glycolylneuraminic acid
(NeuGc) [43]. Additionally, structures included a tetrasaccharide core containing three hexose (e.g.,
mannose) moieties and one HexNAc (N-acetylglucosamine) moiety. Structures were confirmed using
MS/MS fragmentation patterns and isotopic distribution. A library of glycan structures specific to the
sample set was assembled based on retention time, molecular formula and confirmed m/z. Relative
abundances of N-glycans were obtained using Masshunter Profinder software (version B.06.00, Agilent
Technologies). The batch targeted feature extraction algorithm with the following parameters was
applied using the generated database: minimum abundance of 750 counts, charge state up to +3,
retention time window of 2 min, and a glycan isotope model. Peak areas were verified for accurate
integration and isotopic distribution. 3.9. Release of N-glycans from Bovine Colostrum whey Proteins by Free and Immobilized Endo Bovine colostrum whey proteins were isolated from bovine colostrum whey kindly provided
by La Belle (Bellingham, WA, USA). Bovine colostrum whey proteins were isolated and purified
using a 10 kDa molecular weight cut-off ultrafiltration as described previosly by Karav et al. [33] and
were deglycosylated by free and immobilized Endo BI-1 in triplicate. Bovine colostrum whey protein
concentration was determined by Qubit fluorometric kit according to manufacturer’s instructions as
described previously. Briefly, Endo BI-1 (free or immobilized) was added to purified colostrum whey
proteins at a ratio of 1:75 in 20 mM Na2HPO4 buffer at pH 7 and reacted for 1 h at 45 ◦C based on
previous reports [33]. The reaction was terminated, and proteins were precipitated using cold ethanol. The supernatant was dried by vacuum centrifuge and subsequently resuspended, whereupon glycans
were purified from contaminants using C18 cartridges (Glygen, Columbia, MD, USA) and desalted by
porous graphitized carbon (PGC, Glygen) solid phase extraction as described previously [7,50]. Eluate
from PGC was dried, resuspended in nanopure water for subsequent nano-LC-Chip Q ToF analysis. 3.8. Evaluating Reusability of Immobilized Endo BI-1 Using RNase B Reusability of immobilized Endo BI-1 was evaluated for ten cycles of 20 min at both 65 ◦C and
45 ◦C in presence of RNase B. Between each cycle, the supernatant from the reaction was removed
and subsequently treated with cold ethanol for protein precipitation and glycan isolation. After each
reaction cycle, the remaining resin containing the immobilized enzyme was washed with a 20 mM
Na2HPO4 buffer at a 4:1 ratio (buffer/reaction volume) and fresh substrate-containing buffer was 12 of 15 Catalysts 2018, 8, 278 12 of 15 added for the next reaction cycle. Enzyme activity after the first reaction was considered as 100%. Experiments were conducted in triplicate. 3.9. Release of N-glycans from Bovine Colostrum whey Proteins by Free and Immobilized Endo BI-1 4. Conclusions These findings highlight the advantages and limitations of different enzyme immobilization strategies,
in particular when considering an enzyme targeting the release of glycans from macromolecular substrates
such as glycoproteins. The immobilization process shifted the optimum pH of the enzyme to a neutral
pH, which is of relevance when considering the immobilization of this enzyme for industrial applications,
such as in dairy products or other food products having a pH close to neutral. The use of moderate
temperatures favored the reuse of the immobilized enzyme. Immobilized enzyme reusability may
compensate for its initial reduced activity. When using a complex and diverse source of glycoproteins as
bovine colostrum, the immobilized enzyme released a higher abundance of neutral N-glycans than the free Catalysts 2018, 8, 278 13 of 15 enzyme, which yielded a different overall composition of glycans by class. This difference suggests that
specific glycan release can be achieved by potentially tailoring immobilized enzyme reactions conditions in
future studies. Further investigation of the different rates of glycan release, possibly due to limited enzyme
accessibility to specific glycan sites on glycoproteins, may enhance our understanding of the role of
certain glycosylation sites and structures on their function, in particular for therapeutic glycoproteins
such as monoclonal antibodies. Investigation on alternative glycoprotein substrates of plant origin is
warranted to understand substrate specificity. This study presents a scalable technique for enzymatic
deglycosylation of industrially relevant glycoproteins. Optimized reactor configurations, including
flow-through column reactors and fluidized beds, could favor glycan release and subsequent isolation
from deglycosylated proteins in downstream isolation, enabling future processing scale-up and
commercialization of bioactive glycans and novel therapeutics. Supplementary Materials: The following are available online at http://www.mdpi.com/2073-4344/8/7/278/s1,
Figure S1: Sodium dodecyl sulfate polyacrylamide gel electrophoresis of immobilization washings from amino,
adsorption, and epoxy methods on a 4–15% acrylamide gel. Endo BI-1 weighs approximately 53 kDa, Figure S2:
Normalized MALDI-ToF MS intensity vs. N glycan: I.S. ratio for glycans released from RNase B, Table S1: Mean
relative quantities (±S.D.) of N-glycans released by free and immobilized Endo BI-1 from bovine colostrum whey
proteins as determined by nano-LC Chip Q-ToF MS/MS Compound formula is signified by the number of residues of
each monosaccharide in the following order: Hex_HexNAc_Fuc_NeuAc_NeuGc. Author Contributions: D.B. and J.M.L.N.de.M.B. came up with the original idea. J.L.C. and J.M.L.N.de.M.B. designed
the experiments. S.K. expressed and isolated the enzyme. J.L.C. performed the experiments and analyzed the data. J.L.C., S.K., D.B., and J.M.L.N.de.M.B. 4. Conclusions wrote the paper. Acknowledgments: This research was supported by the Center for Advanced Processing and Packaging Studies
(CAPPS), the National Institutes of Health awards R01AT007079 and R01AT008759, the USDA:NIFA Hatch project
232719, and was partially supported by an industry/campus supported fellowship under the Training Program in
Biomolecular Technology (T32-GM008799) at the University of California, Davis. The authors would like to thank
David Mills for providing the endo- β-N-acetylglucosaminidase enzyme and Nannan Tao from Bruker Daltonics
for assistance with MALDI-ToF MS applications. Conflicts of Interest: D.B. is a cofounder of Evolve Biosystems, a company focused on diet-based manipulation
of the gut microbiota. Evolve Biosystems played no role in the funding, design, execution, interpretation,
or publication of this work. The other authors declare no conflict of interest. References [CrossRef] Catalysts 2018, 8, 278 14 of 15 14 of 15 9. Garrido, D.; Nwosu, C.; Ruiz-Moyano, S.; Aldredge, D.; German, J.B.; Lebrilla, C.B.; Mills, D.A. Endo-β-N-
acetylglucosaminidases from Infant Gut-associated Bifidobacteria Release Complex N-glycans from Human
Milk Glycoproteins. Mol. Cell. Proteom. 2012, 11, 775–785. [CrossRef] [PubMed] 10. Wang, W.-L.; Wang, W.; Du, Y.-M.; Wu, H.; Yu, X.-B.; Ye, K.-P.; Li, C.-B.; Jung, Y.-S.; Qian, Y.-J.; Voglmeir, J.; et al. Comparison of anti-pathogenic activities of the human and bovine milk N-glycome: Fucosylation is a key factor. Food Chem. 2017, 235, 167–174. [CrossRef] [PubMed] 11. Blaschek, K.M.; Wendorff, W.L.; Rankin, S.A. Survey of Salty and Sweet Whey Composition from Various
Cheese Plants in Wisconsin. J. Dairy Sci. 2007, 90, 2029–2034. [CrossRef] [PubMed] 12. Walstra, P.; Walstra, P.; Wouters, J.T.M.; Geurts, T.J. Dairy Science and Technology, 2nd ed.; CRC Press:
Boca Raton, Fl, USA, 2010; ISBN 978-1-4200-2801-0. 13. USDA. Dairy Products 2016 Summary; USDA National Agricultural Statistics Service, United States Department
of Agriculture: Washington, DC, USA, 2017. 14. Cao, L.; van Langen, L.; Sheldon, R.A. Immobilised enzymes: Carrier-bound or carrier-free? Curr. Opin. Biotechnol. 2003, 14, 387–394. [CrossRef] 5. Cao, L. Immobilised enzymes: science or art? Curr. Opin. Chem. Biol. 2005, 9, 217–226. [CrossRef] [PubM 15. Cao, L. Immobilised enzymes: science or art? Curr. Opin. Chem. Biol. 2005, 9, 217–226. [CrossRef] [PubMed]
16. DiCosimo, R.; McAuliffe, J.; Poulose, A.J.; Bohlmann, G. Industrial use of immobilized enzymes. Chem. Soc. Rev. 2013, 42, 6437–6474. [CrossRef] [PubMed] 16. DiCosimo, R.; McAuliffe, J.; Poulose, A.J.; Bohlmann, G. Industrial use of immobilized enzymes. Chem. Soc. Rev. 2013, 42, 6437–6474. [CrossRef] [PubMed] 17. Fernandes, P. Enzymes in Food Processing: A Condensed Overview on Strategies for Better Biocatalysts. Enzyme Res. 2010, 2010, 862537. [CrossRef] [PubMed] 18. Alloue, W.A.M.; Destain, J.; Medjoub, T.E.; Ghalfi, H.; Kabran, P.; Thonart, P. Comparison of Yarrowia
lipolytica Lipase Immobilization Yield of Entrapment, Adsorption, and Covalent Bond Techniques. Appl. Biochem. Biotechnol. 2008, 150, 51–63. [CrossRef] [PubMed] 19. Mateo, C.;
Grazú, V.;
Pessela, B.C.C.;
Montes, T.;
Palomo, J.M.;
Torres, R.;
López-Gallego, F.;
Fernández-Lafuente, R.; Guisán, J.M. Advances in the design of new epoxy supports for enzyme
immobilization–stabilization. Biochem. Soc. Trans. 2007, 35, 1593–1601. [CrossRef] [PubMed] 20. Karav, S.; Le Parc, A.; de Moura Bell, J.M.L.N.; Rouquié, C.; Mills, D.A.; Barile, D.; Block, D.E. Others Kinetic
characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to
release bioactive glycans. Enzyme Microb. Technol. 2015, 77, 46–53. References 1. Lafite, P.; Daniellou, R. Rare and unusual glycosylation of peptides and proteins. Nat. Prod. Rep. 2012, 29,
729–738. [CrossRef] [PubMed] 1. Lafite, P.; Daniellou, R. Rare and unusual glycosylation of peptides and proteins. Nat. Prod. Rep. 2012, 29,
729–738. [CrossRef] [PubMed] 2. Montreuil, J. Primary Structure of Glycoprotein Glycans Basis for the Molecular Biology of Glycoproteins. In Advances in Carbohydrate Chemistry and Biochemistry; Elsevier: Cambridge, MA, USA, 1980; Volume 37,
pp. 157–223, ISBN 978-0-12-007237-8. 2. Montreuil, J. Primary Structure of Glycoprotein Glycans Basis for the Molecular Biology of Glycoproteins. In Advances in Carbohydrate Chemistry and Biochemistry; Elsevier: Cambridge, MA, USA, 1980; Volume 37,
pp. 157–223, ISBN 978-0-12-007237-8. 3. Zivkovic, A.M.; German, J.B.; Lebrilla, C.B.; Mills, D.A. Human milk glycobiome and its impact on the infant
gastrointestinal microbiota. Proc. Natl. Acad. Sci. USA 2011, 108, 4653–4658. [CrossRef] [PubMed] 3. Zivkovic, A.M.; German, J.B.; Lebrilla, C.B.; Mills, D.A. Human milk glycobiome and its impact on the infant
gastrointestinal microbiota. Proc. Natl. Acad. Sci. USA 2011, 108, 4653–4658. [CrossRef] [PubMed] 4. Thurl, S.; Munzert, M.; Henker, J.; Boehm, G.; Müller-Werner, B.; Jelinek, J.; Stahl, B. Variation of human
milk oligosaccharides in relation to milk groups and lactational periods. Br. J. Nutr. 2010, 104, 1261–1271. [CrossRef] [PubMed] 4. Thurl, S.; Munzert, M.; Henker, J.; Boehm, G.; Müller-Werner, B.; Jelinek, J.; Stahl, B. Variation of human
milk oligosaccharides in relation to milk groups and lactational periods. Br. J. Nutr. 2010, 104, 1261–1271. [CrossRef] [PubMed] . Helenius, A.; Aebi, M. Intracellular Functions of N-Linked Glycans. Science 2001, 291, 2364–2369. [Cross
[PubMed] 5. Helenius, A.; Aebi, M. Intracellular Functions of N-Linked Glycans. Science 2001, 291, 2364–2369. [CrossRef]
[PubMed] [PubMed]
6. Karav, S.; Cohen, J.L.; Barile, D.; de Moura Bell, J.M.L.N. Recent advances in immobilization strategies for
glycosidases. Biotechnol. Prog. 2017, 33, 104–112. [CrossRef] [PubMed] . Karav, S.; Cohen, J.L.; Barile, D.; de Moura Bell, J.M.L.N. Recent advances in immobilization strategies
glycosidases. Biotechnol. Prog. 2017, 33, 104–112. [CrossRef] [PubMed] 7. Karav, S.; Parc, A.L.; de Moura Bell, J.M.L.N; Frese, S.A.; Kirmiz, N.; Block, D.E.; Barile, D.; Mills, D.A. Oligosaccharides Released from Milk Glycoproteins Are Selective Growth Substrates for Infant-Associated
Bifidobacteria. Appl. Environ. Microbiol. 2016, 82, 3622–3630. [CrossRef] [PubMed] 8. Wang, W.L.; Du, Y.M.; Wang, W.; Conway, L.P.; Cai, Z.P.; Voglmeir, J.; Liu, L. Comparison of the bifidogenic
activity of human and bovine milk N-glycome. J. Funct. Foods 2017, 33, 40–51. References [CrossRef] [PubMed] g y
y
21. Masuko, T.; Minami, A.; Iwasaki, N.; Majima, T.; Nishimura, S.-I.; Lee, Y.C. Carbohydrate analysis by a
phenol–sulfuric acid method in microplate format. Anal. Biochem. 2005, 339, 69–72. [CrossRef] [PubMed] 21. Masuko, T.; Minami, A.; Iwasaki, N.; Majima, T.; Nishimura, S.-I.; Lee, Y.C. Carbohydrate analysis by a
phenol–sulfuric acid method in microplate format. Anal. Biochem. 2005, 339, 69–72. [CrossRef] [PubMed]
22. Scott, T.A., Jr.; Melvin, E.H. Determination of dextran with anthrone. Anal. Chem. 1953, 25, 1656–1661. [
f] phenol–sulfuric acid method in microplate format. Anal. Biochem. 2005, 339, 69–72. [CrossRef] [PubMed]
22. Scott, T.A., Jr.; Melvin, E.H. Determination of dextran with anthrone. Anal. Chem. 1953, 25, 1656–1661. [CrossRef] 22. Scott, T.A., Jr.; Melvin, E.H. Determination of dextran with anthrone. Anal. Chem. 1953, 25, 1656–1661. [CrossRef] 23. Pfenninger, A.; Karas, M.; Finke, B.; Stahl, B.; Sawatzki, G. Mass Spectrometric Investigations of Human Milk
Oligosaccharides. In Bioactive Components of Human Milk; Advances in Experimental Medicine and Biology;
Springer: Boston, MA, USA, 2001; pp. 279–284. ISBN 978-1-4613-5521-2. 24. Nishimura, S.-I.; Niikura, K.; Kurogochi, M.; Matsushita, T.; Fumoto, M.; Hinou, H.; Kamitani, R.;
Nakagawa, H.; Deguchi, K.; Miura, N.; et al. High-Throughput Protein Glycomics: Combined Use of
Chemoselective Glycoblotting and MALDI-TOF/TOF Mass Spectrometry. Angew. Chem. Int. Ed. 2005, 44,
91–96. [CrossRef] [PubMed] 25. Bucknall, M.; Fung, K.Y.C.; Duncan, M.W. Practical quantitative biomedical applications of MALDI-TOF
mass spectrometry. J. Am. Soc. Mass Spectrom. 2002, 13, 1015–1027. [CrossRef] 26. Bothner, B.; Chavez, R.; Wei, J.; Strupp, C.; Phung, Q.; Schneemann, A.; Siuzdak, G. Monitoring Enzyme
Catalysis with Mass Spectrometry. J. Biol. Chem. 2000, 275, 13455–13459. [CrossRef] [PubMed] 27. Leboeuf, E.; Immerzeel, P.; Gibon, Y.; Steup, M.; Pauly, M. High-throughput functional assessment
of polysaccharide-active enzymes using matrix-assisted laser desorption/ionization–time-of-flight mass
spectrometry as exemplified on plant cell wall polysaccharides. Anal. Biochem. 2008, 373, 9–17. [CrossRef]
[PubMed] 28. Kang, M.-J.; Tholey, A.; Heinzle, E. Application of automated matrix-assisted laser desorption/ionization
time-of-flight mass spectrometry for the measurement of enzyme activities. Rapid Commun. Mass Spectrom. 2001, 15, 1327–1333. [CrossRef] [PubMed] 29. Mechref, Y.; Novotny, M.V.; Krishnan, C. Structural Characterization of Oligosaccharides Using Maldi-TOF/
TOF Tandem Mass Spectrometry. Anal. Chem. 2003, 75, 4895–4903. [CrossRef] [PubMed] 15 of 15 15 of 15 Catalysts 2018, 8, 278 30. Gaur, R.; Pant, H.; Jain, R.; Khare, S.K. Galacto-oligosaccharide synthesis by immobilized Aspergillus oryzae
β-galactosidase. Food Chem. 2006, 97, 426–430. [CrossRef] 31. Wong, D.E.; Senecal, K.J.; Goddard, J.M. References Immobilization of chymotrypsin on hierarchical nylon 6,6 nanofiber
improves enzyme performance. Colloids Surf. B Biointerfaces 2017, 154, 270–278. [CrossRef] [PubMed] 32. Rehman, H.U.; Aman, A.; Silipo, A.; Qader, S.A.U.; Molinaro, A.; Ansari, A. Degradation of complex
carbohydrate: Immobilization of pectinase from Bacillus licheniformis KIBGE-IB21 using calcium alginate as
a support. Food Chem. 2013, 139, 1081–1086. [CrossRef] [PubMed] 33. Karav, S.; de Moura Bell, J.M.L.N.; Parc, A.L.; Liu, Y.; Mills, D.A.; Block, D.E.; Barile, D. Characterizing
the release of bioactive N-glycans from dairy products by a novel endo-β-N-acetylglucosaminidase. Biotechnol. Prog. 2015, 31, 1331–1339. [CrossRef] [PubMed] 34. Li, D.; Teoh, W.Y.; Gooding, J.J.; Selomulya, C.; Amal, R. Functionalization Strategies for Protease
Immobilization on Magnetic Nanoparticles. Adv. Funct. Mater. 2010, 20, 1767–1777. [CrossRef] 35. Iyer, P.V.; Ananthanarayan, L. Enzyme stability and stabilization—Aqueous and non-aqueous environment. Process Biochem. 2008, 43, 1019–1032. [CrossRef] 36. Busto, M.D.; García-Tramontín, K.E.; Ortega, N.; Perez-Mateos, M. Preparation and properties of an
immobilized pectinlyase for the treatment of fruit juices. Bioresour. Technol. 2006, 97, 1477–1483. [CrossRef]
[PubMed] 37. Demirel, G.; Özçetin, G.; ¸Sahin, F.; Tümtürk, H.; Aksoy, S.; Hasirci, N. Semi-interpenetrating polymer
networks (IPNs) for entrapment of glucose isomerase. React. Funct. Polym. 2006, 66, 389–394. [CrossRef]
38. Wei, L.; Zhang, W.; Lu, H.; Yang, P. Immobilization of enzyme on detonation nanodiamond for highly
ffii
t
t
l
i
T l
t 2010 80 1298 1304 [C
R f] [P bM d] 37. Demirel, G.; Özçetin, G.; ¸Sahin, F.; Tümtürk, H.; Aksoy, S.; Hasirci, N. Semi-interpenetrating polymer
networks (IPNs) for entrapment of glucose isomerase. React. Funct. Polym. 2006, 66, 389–394. [CrossRef] 38. Wei, L.; Zhang, W.; Lu, H.; Yang, P. Immobilization of enzyme on detonation nanodiamond for highly
efficient proteolysis. Talanta 2010, 80, 1298–1304. [CrossRef] [PubMed] 39. Klein, M.P.; Nunes, M.R.; Rodrigues, R.C.; Benvenutti, E.V.; Costa, T.M.H.; Hertz, P.F.; Ninow, J.L. Effect of the
Support Size on the Properties of β-Galactosidase Immobilized on Chitosan: Advantages and Disadvantages
of Macro and Nanoparticles. Biomacromolecules 2012, 13, 2456–2464. [CrossRef] [PubMed] 40. Chauhan, S.; Vohra, A.; Lakhanpal, A.; Gupta, R. Immobilization of Commercial Pectinase (Polygalacturonase)
on Celite and Its Application in Juice Clarification. J. Food Process. Preserv. 2015, 6, 2135–2141. [CrossRef] 41. Parc, A.L.; Karav, S.; de Moura Bell, J.M.L.N.; Frese, S.A.; Liu, Y.; Mills, D.A.; Block, D.E.; Barile, D. A novel
endo-β-N-acetylglucosaminidase releases specific N-glycans depending on different reaction conditions. Biotechnol. Prog. 2015, 31, 1323–1330. [CrossRef] [PubMed] g
42. Axelsson, A.; Zacchi, G. References Economic evaluation of the hydrolysis of lactose using immobilized β-galactosidase. Appl. Biochem. Biotechnol. 1990, 24, 679–693. [CrossRef] 43. Nwosu, C.C.; Aldredge, D.L.; Lee, H.; Lerno, L.A.; Zivkovic, A.M.; German, J.B.; Lebrilla, C.B. Comparison of
the Human and Bovine Milk N-glycome via High-Performance Microfluidic Chip Liquid Chromatography
and Tandem Mass Spectrometry. J. Proteome Res. 2012, 11, 2912–2924. [CrossRef] [PubMed] 44. Huang, Y.; Orlando, R. Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F:
All Glycans Are Not Created Equal. J. Biomol. Tech. JBT 2017. [CrossRef] [PubMed] 45. Tanaka, H.; Matsumura, M.; Veliky, I.A. Diffusion characteristics of substrates in Ca-alginate gel beads. Biotechnol. Bioeng. 1984, 26, 53–58. [CrossRef] [PubMed] 46. Manjon, A.; Iborra, J.L.; Lozano, P.; Canovas, M. A practical experiment on enzyme immobilization and
characterization of the immobilized derivatives. Biochem. Mol. Biol. Educ. 1995, 23, 213–216. [CrossRef]
47. Park, E.; Yang, H.; Kim, Y.; Kim, J. Analysis of oligosaccharides in beer using MALDI-TOF-MS. Food Chem. 2012, 134, 1658–1664. [CrossRef] [PubMed] 46. Manjon, A.; Iborra, J.L.; Lozano, P.; Canovas, M. A practical experiment on enzyme immobilization and
characterization of the immobilized derivatives. Biochem. Mol. Biol. Educ. 1995, 23, 213–216. [CrossRef] 7. Park, E.; Yang, H.; Kim, Y.; Kim, J. Analysis of oligosaccharides in beer using MALDI-TOF-MS. Food C
2012, 134, 1658–1664. [CrossRef] [PubMed] 48. Lineweaver, H.; Burk, D. The determination of enzyme dissociation constants. J. Am. Chem. Soc. 1934, 56,
658–666. [CrossRef] 49. Hanes, C.S. Studies on plant amylases: The effect of starch concentration upon the velocity of hydrolysis by
the amylase of germinated barley. Biochem. J. 1932, 26, 1406–1421. [CrossRef] [PubMed] 50. Cohen, J.L.; Barile, D.; Liu, Y.; de Moura Bell, J.M.L.N. Role of pH in the recovery of bovine milk
oligosaccharides from colostrum whey permeate by nanofiltration. Int. Dairy J. 2017, 66, 68–75. [CrossRef]
[PubMed] © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).
|
https://openalex.org/W2043768713
|
https://www.scielo.br/j/fm/a/SDQTXdJD3NSyLjpjV9zck9b/?lang=pt&format=pdf
|
Portuguese
| null |
Desenvolvimento de um modelo de pé segmentado para avaliação de indivíduos calçados
|
Fisioterapia em Movimento
| 2,013
|
cc-by
| 6,238
|
Development of a segmented foot model for
assessing individuals with shoes Renan Alves Resende[a], Lucas Rodrigues Nascimento[a], Maria Clarice Lopes Silva[b],
Ana Cisalpino Pinheiro[b], Sérgio Teixeira Fonseca[c], Renata Noce Kirkwood[d] [a] Doutorandos em Ciências da Reabilitação da Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG - Brasil,
e-mails: renanresende@hotmail.com, lrn@ufmg.br [a] Doutorandos em Ciências da Reabilitação da Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG - Brasil,
e-mails: renanresende@hotmail.com, lrn@ufmg.br
[b] Acadêmicas do Curso de Fisioterapia da Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG - Brasil,
e-mails: mcla.lopes@yahoo.com.br, anacisalpino@gmail.com
[c] Professor associado do Departamento de Fisioterapia da Universidade Federal de Minas Gerais (UFMG), Belo Horizonte,
MG - Brasil, e-mail: sfonseca@eeffto.ufmg.br
[d] Professora adjunta do Departamento de Fisioterapia da Universidade Federal de Minas Gerais (UFMG), Belo Horizonte,
MG - Brasil, e-mail: renata.kirkwood@gmail.com [b] Acadêmicas do Curso de Fisioterapia da Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG - Brasil,
e-mails: mcla.lopes@yahoo.com.br, anacisalpino@gmail.com [d] Professora adjunta do Departamento de Fisioterapia da Universidade Federal de Minas Gerais (UFMG), Belo Horizonte,
MG - Brasil, e-mail: renata.kirkwood@gmail.com ISSN 0103-5150
Fisioter. Mov., Curitiba, v. 26, n. 1, p. 95-105, jan./mar. 2013
Licenciado sob uma Licença Creative Commons Abstract Introduction: The use of shoes may influence the kinematics parameters of the foot segments during gait. Objective: To test a biomechanical model developed to measure the angular displacement of the foot segments
during gait of individuals using shoes. Materials and methods: Ten healthy young individuals participated in
the study. A pair of shoes was used during the study. The Qualisys Pró-Reflex system was used to measure the fol-
lowing outcome variables: angular displacement of the rearfoot in the sagittal, frontal and transverse planes of
motion and forefoot in the frontal and transverse planes. The Intraclass Correlation Coefficient was calculated to
verify the association between the following variables between visits: maximum and minimum joint angles and
the total range of motion. Results: The investigated outcomes demonstrated values of intra-rater reliability rang-
ing from moderate to excellent. Discussion: This study presented a feasible and reliable method to measure the
angular displacement of the foot segments during gait using shoes. The methods can be applied during scientific
researches and clinical assessments of the kinematics during human gait. Conclusion: The quality of the method
developed and the reliability analysis realized demonstrated motion patterns of the segments of the foot similar
to what is demonstrated in the literature. In addition, the protocol is noninvasive, does not require an additional
reference device, subtalar neutral position and does not depend on information provided by X-ray images. [K] Resumo Introdução: O uso de calçados pode influenciar parâmetros cinemáticos dos segmentos do pé durante a
marcha. Objetivo: Testar um modelo biomecânico para avaliar o deslocamento angular dos segmentos do
pé durante a marcha de indivíduos calçados. Materiais e métodos: Dez indivíduos adultos jovens saudáveis
participaram do estudo. Um par de tênis foi utilizado durante o estudo. O sistema Qualisys Pró-Reflex foi utili-
zado para a avaliação das variáveis de desfecho: deslocamento angular de retropé nos planos sagital, frontal e
transverso e de antepé nos planos frontal e transverso. O Coeficiente de Correlação Intraclasse foi usado para
verificar o grau de associação das seguintes variáveis entre as duas visitas: valores angulares máximos e mí-
nimos e deslocamento angular total. Resultados: As variáveis de desfecho apresentaram confiabilidade teste-
-reteste de moderada a excelente. Discussão: O nível de confiabilidade encontrado foi considerado aceitável,
fornecendo, assim, uma base sólida para a avaliação objetiva da cinemática dos segmentos do pé de indivíduos
calçados em ambientes laboratoriais, para fins de pesquisas científicas e avaliações clínicas da cinemática da Fisioter Mov. 2013 jan/mar;26(1):95-105 Resende RA, Nascimento LR, Silva MCL, Pinheiro AC, Fonseca ST, Kirkwood RN. 96 marcha humana. Conclusão: A qualidade do método desenvolvido e da análise de confiabilidade realizada
detectou padrões de movimento dos segmentos do pé próximos aos descritos na literatura. Além disso, o pro-
tocolo de aquisição apresentado não é invasivo, não requer uma estrutura extra de referência, posicionamento
da articulação subtalar em neutro nem é dependente da informação fornecida por raios-X. [P] marcha humana. Conclusão: A qualidade do método desenvolvido e da análise de confiabilidade realizada
detectou padrões de movimento dos segmentos do pé próximos aos descritos na literatura. Além disso, o pro-
tocolo de aquisição apresentado não é invasivo, não requer uma estrutura extra de referência, posicionamento
da articulação subtalar em neutro nem é dependente da informação fornecida por raios-X. [P] Palavras-chave: Cinemática. Pé. Marcha. Confiabilidade. Calçados. Fisioter Mov. 2013 jan/mar;26(1):95-105 Keywords: Kinematics. Foot. Gait. Reliability. Shoes. Introdução A perna e o complexo do pé são importantes
componentes do sistema músculo-esquelético res-
ponsáveis pela interação entre o membro inferior e a
superfície de apoio (1) permitindo, dessa forma, a ade-
quada distribuição das cargas inerciais e gravitacio-
nais durante a marcha humana (2). Anormalidades no
pé ou tornozelo podem causar distribuição irregular
das cargas impostas ao sistema musculoesquelético e
determinar adaptações compensatórias que impõem
demandas excessivas em diversas estruturas do mem-
bro inferior, favorecendo o surgimento de patologias
insidiosas, inclusive em regiões proximais (3, 4). dos movimentos das articulações do complexo do pé
nos demais segmentos do membro inferior e refor-
çam a importância de uma adequada avaliação desses
movimentos durante a análise de marcha. Embora os movimentos do pé apresentem relativa
especificidade e complexidade devido à eminente
característica multiplanar das superfícies de contato
das articulações do pé, os modelos biomecânicos clí-
nicos utilizados usualmente representam o pé como
um vetor rígido único, o que permite que apenas os
ângulos de progressão do pé de flexão plantar e de
dorsiflexão sejam determinados (8, 9). Tais modelos
são insuficientes para a caracterização dos movimen-
tos do pé em três dimensões durante a fase de apoio
da marcha (10). Evidências científicas indicaram que corredores
classificados como pronadores excessivos apresen-
tam maior valgismo dinâmico de joelho durante a fase
de contato inicial o que poderia levar a uma maior
sobrecarga sobre o compartimento medial do joelho
(5). Nesse contexto, a eversão excessiva do calcâneo
durante a fase de apoio da marcha é usualmente men-
cionada como um dos possíveis fatores determinan-
tes de modificações no padrão cinemático de joelho e
quadril (6, 7). Tais achados demonstram a influência Carson et al. (8) desenvolveram um modelo bio-
mecânico de pé segmentado associado a um protoco-
lo de avaliação aplicável à análise da marcha. Foram
demonstrados padrões de movimento das diferen-
tes partes do pé consistentes com o conhecimento
existente acerca da cinemática do pé e as medidas
apresentaram confiabilidade aceitável, o que somado Desenvolvimento de um modelo de pé segmentado para avaliação de indivíduos calçados 97 a outros modelos previamente descritos (2, 8, 11),
forneceu uma base sólida para a avaliação objetiva
dos movimentos do pé. Análise Cinemática As variáveis cinemáticas de desfecho do estudo
foram valores máximo e mínimo e amplitude de
movimento (ADM) total nos planos frontal e trans-
verso de antepé em relação ao retropé e nos planos
sagital, frontal e transverso do retropé em relação
à perna durante a fase de apoio da marcha. Cada
indivíduo participou de duas coletas, com interva-
lo de sete dias. Esse procedimento foi necessário
para permitir a verificação do grau de associação
entre os dados a partir do cálculo dos coeficientes
de confiabilidade teste-reteste. Para tanto, um modelo que seja capaz de avaliar os
diferentes segmentos do pé durante a marcha com os
indivíduos calçados deve ser desenvolvido e testado
para garantir a reprodutibilidade de suas medidas. Dessa forma, o objetivo do presente estudo foi desen-
volver um modelo biomecânico composto por per-
na, retropé e antepé para avaliação do deslocamento
angular dos segmentos do pé durante a marcha com
indivíduos calçados e avaliar a confiabilidade teste-
-reteste das medidas obtidas. O deslocamento angular de antepé em relação
ao retropé e retropé em relação à perna foi obti-
do com o uso do sistema de análise de movimento
Qualisys – Pro Reflex MCU (QUALISYS MEDICAL AB,
411 12 Gothenburg, Suécia). A coleta envolveu o uso
de oito câmeras com iluminação produzida por um
grupo de diodos emissores de luz infravermelha, lo-
calizados em volta da lente de cada uma das câme-
ras. O Sistema Qualisys permite a reconstrução em
três dimensões de marcadores passivos refletores
localizados em proeminências ósseas especificas. Os
marcadores passivos refletores captam e refletem a
luz infravermelha, que é então captada pela lente das
câmeras. Foi utilizada a frequência de coleta de 120 Hz. Sincronizado ao sistema Qualisys, uma plataforma
de força AMTI® (Advanced Mechanical Tecnology,
modelo OR6-6, Watertown, MA, USA) embutida na
passarela foi utilizada para captar os dados da força
de reação de solo necessários para a delimitação da
fase de apoio da marcha. A calibração foi realizada de
acordo com o manual do fabricante e foram permiti-
dos erros de desvio padrão menores que 10 mm (14). Introdução alterações musculares ou neurológicas que interfi-
ram no desempenho da marcha; (vi) aceitar partici-
par da pesquisa e assinar o Termo de Consentimento
Livre e Esclarecido (TCLE), previamente aprovado
pelo Comitê de Ética em Pesquisa da UFMG: ETIC
0047.0.203.000-10. Os indivíduos foram excluídos
quando houve relato de desconforto durante qual-
quer procedimento da coleta de dados. Embora a maior parte da população faça uso regu-
lar de calçados durante a realização de suas ativida-
des rotineiras, os modelos descritos para a avaliação
dos movimentos do pé usualmente avaliam os indi-
víduos deambulando sem o uso de calçados, o que
pode influenciar significativamente os parâmetros
de avaliação ou mesmo os parâmetros cinemáticos
da marcha humana. Tais avaliações podem, portanto,
determinar resultados que não refletem, da melhor
maneira, a movimentação usual dos segmentos do
pé, sendo que a avaliação desses segmentos com o
indivíduo calçado poderá fornecer informações mais
fidedignas acerca do padrão cinemático do pé no dia
a dia (12, 13). Delineamento do estudo Foi realizado um estudo do tipo metodológico no
Laboratório de Análise de Movimento da Escola de
Educação Física, Fisioterapia e Terapia Ocupacional
da Universidade Federal de Minas Gerais (UFMG). Fisioter Mov. 2013 jan/mar;26(1):95-105 Amostra A amostra do tipo conveniência foi composta
por voluntários jovens, recrutados na comunidade
de acordo com os seguintes critérios de inclusão: (i)
ter idade entre 18 e 30 anos; (ii) utilizar no dia a dia
tênis com tamanhos entre 37 e 39 (medida brasilei-
ra); (iii) não ter sofrido lesões ou ter sido submetido
a qualquer tipo de cirurgia de membros inferiores;
(iv) não apresentar deficit visuais significativos não
corrigidos por lentes corretivas; (v) não possuir Para a construção do modelo biomecânico
composto por antepé, retropé e perna, marcado-
res reflexivos (9 mm) foram afixados nos epicôn-
dilos lateral e medial do fêmur, maléolo lateral e
medial, sustentáculo do tálus, tróclea fibular, base Fisioter Mov. 2013 jan/mar;26(1):95-105 Resende RA, Nascimento LR, Silva MCL, Pinheiro AC, Fonseca ST, Kirkwood RN. 98 e cabeça do primeiro e quinto metatarsos, estando
inicialmente o indivíduo descalço. Para possibilitar
o rastreamento dos segmentos durante a marcha,
clusters foram fixados com faixa elástica em regiões
específicas: (i) um cluster semirrígido composto por
três marcadores refletores foi posicionado no ter-
ço médio da perna, (ii) um cluster rígido com três
marcadores foi posicionado sobre o calcâneo, pró-
ximo à inserção do tendão calcâneo e, no segmento
antepé, (iii) um cluster rígido com três marcadores
foi posicionado sobre a região correspondente às
diáfises dos segundo e terceiro metatarsos. Uma
coleta estática foi conduzida para o registro da posi-
ção inicial e orientação dos marcadores anatômicos
e de rastreamento. A coleta estática foi realizada
com o indivíduo descalço em posição ortostática,
o que possibilitou a utilização dos marcadores de
definição e de rastreamento dos segmentos do pé
e da perna durante a coleta (Figura 1). A palpação
das proeminências ósseas e fixação dos marcadores foram realizadas sempre pelo mesmo examinador, o
qual possuía experiência com o processo de coleta
de dados. Todos os ângulos reportados foram rela-
tivos a essa posição estática de referência. Após a coleta estática, os marcadores de epicôn-
dilos lateral e medial do fêmur, maléolo lateral e
medial, sustentáculo do tálus, tróclea fibular, base e
cabeça do 1º e 5º metatarsos foram retirados para
que pudessem ser realizadas as coletas dinâmicas. Um avaliador previamente treinado foi responsável
por retirar os marcadores iniciais e calçar o tênis nos
participantes sem que houvesse movimento dos clus-
ters de rastreamento de perna, retropé e antepé. Amostra Após
a colocação dos tênis, foi permitido um período de
cinco minutos para que o indivíduo se adaptasse ao
tênis, à passarela e aos marcadores; após esse perío-
do, iniciaram-se as coletas dinâmicas nas quais um
mínimo de cinco passadas em velocidade autossele-
cionada foi coletado em uma passarela de dez metros
de comprimento (Figura 2). Figura 1 - Coleta estática com o indivíduo descalço
Fonte: Dados da pesquisa. Figura 2 - Coleta dinâmica com o indivíduo calçado
Fonte: Dados da pesquisa. Figura 1 - Coleta estática com o indivíduo descalço
Fonte: Dados da pesquisa. Figura 2 - Coleta dinâmica com o indivíduo calçado
Fonte: Dados da pesquisa. Fisioter Mov. 2013 jan/mar;26(1):95-105 Amostra e variáveis espaçotemporais Os ângulos articulares foram calculados usando-se
a sequência de Cardan e definidos a partir da orien-
tação do sistema de coordenadas de um segmento
corporal em relação ao sistema de coordenadas do
segmento de referência. A perna foi utilizada como
segmento referência para o cálculo do ângulo do re-
tropé, e este foi utilizado como segmento referência
para o cálculo do ângulo do antepé. Em seguida, foi
aplicado um filtro passa-baixa Butterworth de quarta
ordem com frequência de corte estabelecida em 6 Hz
(15). Foram gerados os gráficos da média dos deslo-
camentos angulares nos três planos do retropé e nos
planos frontal e transverso de antepé durante a fase
de apoio da marcha. Para a geração dos gráficos, foi
utilizada a convenção de deslocamentos angulares
positivos para dorsiflexão de retropé e inversão e
adução de retropé e antepé. Participaram do estudo dez adultos jovens saudá-
veis (sete do sexo feminino e três do sexo masculino)
com média de idade de 25 anos (DP = 3,3), média de
massa e de altura de 63kg (DP = 8,9) e 1,70 m
(DP = 0,04), respectivamente. Baseado em Walter
et al. (17), dez indivíduos são suficientes para a rea-
lização de um estudo de confiabilidade com um alfa
de 0,05 e beta de 0,2. As velocidades de marcha apresentadas pelos
indivíduos foram similares entre as duas condições
experimentais durante ambas as coletas, permitindo
comparação adequada dos resultados sem influência
da velocidade. Durante a primeira visita, os indivíduos
apresentaram velocidade média de 1,27 m/s (DP =
0,13; IC 95% [1,17-1,37]) e comprimento da passada
de 1,38 m (DP = 0,11; IC 95% [1,30-1,47]); durante
a segunda visita, eles apresentaram 1,37 m/s (DP =
0,12; IC 95% [1,28-1,47]) e 1,46 m (DP = 0,04; IC95%
[1,43-1,50]) de velocidade média e comprimento da
passada, respectivamente. Os dados foram exportados para o programa
Microsoft Excel, onde foram calculados os valores an-
gulares máximos e mínimos e as ADM totais a partir
da subtração do menor valor angular do maior valor
angular durante a fase de apoio da marcha. Características dos tênis utilizados do cluster de retropé posicionado sobre o calcâneo. Essa fixação com velcro era aberta no momento de
calçar o tênis no participante e fechada após a co-
locação do calçado. Após a fixação, a abertura apre-
sentou dimensões de 2,8 cm por 2,0 cm. Essas adap-
tações possibilitaram a manutenção dos clusters de
rastreamento entre a coleta estática e dinâmica sem
modificação da posição inicial dos mesmos com mí-
nima alteração da firmeza proporcionada pelo tênis
durante a marcha. Um par de tênis, tamanho 39 (medida brasilei-
ra), desenvolvido especificamente para a prática de
corrida e caminhada, foi adquirido para a realização
do estudo. O par de tênis teve suas linguetas retira-
das para possibilitar o uso do cluster de antepé e,
em sua parte posterior, foi realizada uma abertura
posteriormente fixada com velcro na parte superior
do talão do tênis com o objetivo de possibilitar o uso Fisioter Mov. 2013 jan/mar;26(1):95-105 Desenvolvimento de um modelo de pé segmentado para avaliação de indivíduos calçados 99 de dispersão com o objetivo de caracterização da
amostra. As médias dos valores máximos, mínimos
e de ADM das duas visitas e a variabilidade intra
e intersujeito foram apresentadas. As medidas de
concordância foram expressas em termos de coefi-
cientes de correlação e a análise estatística dos dados
foi feita utilizando-se o Coeficiente de Correlação
Intraclasse (ICC). Foi calculada a confiabilidade
teste-reteste para as variáveis de desfecho do estu-
do. A força ou magnitude da relação entre as medidas
foi classificada como fraca (ICC < 0,4), moderada
(0,4 ≤ ICC ≤ 0,75) e forte (ICC > 0,75) (16). Na maior
parte dos estudos, valores de confiabilidade maiores
do que 40% são necessários para prover um método
de medida útil clinicamente (17). Todos os cálculos
foram realizados utilizando o programa estatístico
SPSS para Windows versão 15.0, com nível de sig-
nificância estabelecido de α = 0,05. Redução dos dados Os dados cinemáticos foram capturados pelo
software de aquisição Qualisys Track Manager 1.9.2. Nessa etapa, realizou-se a interpolação quando a tra-
jetória dos marcadores foi perdida por no máximo 10
quadros. Foram excluídas as passadas que apresen-
taram algum artefato, como má qualidade do sinal. Todos os dados coletados foram processados por um
segundo examinador, sendo este cegado em relação
aos indivíduos e ao dia da coleta com uso de códigos
associados a cada voluntário. Os dados cinemáticos foram capturados pelo
software de aquisição Qualisys Track Manager 1.9.2. Em seguida, os dados foram exportados para o
software Visual3D C-Motion, Inc para seu processa-
mento, utilizando as rotações padronizadas de Euler
(X-Y-Z). Foram realizadas a construção do modelo
biomecânico dos segmentos corporais e a aplicação
desse modelo nos arquivos dinâmicos. Os dados da
plataforma de força foram processados e utilizados
para a normalização da fase de apoio da marcha,
de 0 a 100% (contato do pé à retirada do mesmo
pé do solo). Fisioter Mov. 2013 jan/mar;26(1):95-105 Antepé-retropé O antepé estava invertido no plano frontal e adu-
zido no plano transverso no contato inicial. Durante
os 10% inicias da fase de apoio, o antepé apresentou
máxima inversão e máxima adução. Eversão e discre-
ta abdução caracterizaram o período entre 10 e 20%
da fase de apoio. Entre 20 e 60%, ocorreram eversão
e abdução. Entre 60 e 80%, ocorreram discreta in-
versão e adução. Na fase final de apoio (80 a 100%),
ocorreram máxima eversão com posterior inversão
e discreta abdução (Gráfico 2). Confi abilidade Os dados antropométricos dos participantes fo-
ram descritos por medidas de tendência central e Os valores angulares máximos e mínimos e as
ADM totais médias das duas visitas e a variabilidade Fisioter Mov. 2013 jan/mar;26(1):95-105 Resende RA, Nascimento LR, Silva MCL, Pinheiro AC, Fonseca ST, Kirkwood RN. 100 intrassujeito e intersujeito foram apresentados no
Quadro 1. A variabilidade intrassujeito foi de 0,99 a
2,37 para ADM, entre 0,77 e 2,29 para valores máxi-
mos e entre 0,77 e 1,53 para valores mínimos. Além
disso, pôde-se observar que as variáveis de desfecho
do estudo apresentaram confiabilidade teste-reteste
variando entre moderada e excelente, e que a men-
suração da ADM total, valor máximo e mínimo do
retropé no plano sagital apresentaram os maiores
valores de ICC entre as variáveis de desfecho do estu-
do. Os valores de ICC para cada variável de desfecho
do estudo foram apresentados no Quadro 2. abdução caracterizaram o período entre 10 e 20%
da fase de apoio. Entre 20 e 60% da fase de apoio,
ocorreram dorsiflexão, inversão e adução do retropé. Entre 60 e 80% da fase de apoio, o retropé moveu-se
para máxima dorsiflexão e permaneceu sem altera-
ções significativas nos planos frontal e transverso. Na fase final de apoio (80 a 100%), ocorreram flexão
plantar, máxima inversão e abdução (Gráfico 1). Retropé-perna No contato inicial, o retropé apresentou-se em
dorsiflexão no plano sagital, discreta inversão no pla-
no frontal e abdução no plano transverso. Durante os
10% inicias da fase de apoio, o retropé moveu-se em
flexão plantar, máxima eversão e abdução. Máxima
flexão plantar e posterior dorsiflexão, inversão e Quadro 1 - Média (em graus) de ADM, valores máximo e mínimo (DP) para as diferentes articulações nos diferentes
planos e variabilidade intrassujeito (média de DP dos indivíduos nas duas visitas)
ADM
Valor máximo
Valor mínimo
Média
Variabilidade
Média
Variabilidade
Média
Variabilidade
Retropé
Sagital
24,8 (7,8)
0,99
35,7 (15,2)
0,77
9,0 (13,2)
0,97
Frontal
10,4 (3,2)
2,37
6,3 (7,1)
2,29
-4,5 (6,9)
1,53
Transverso
13,7 (3,8)
1,14
-14,9 (5,4)
1,11
-28,5 (7,2)
0,91
Antepé
Frontal
6,8 (1,9)
1,32
4,0 (7,0)
1,09
-2,8 (7,5)
1,04
Transverso
4,1 (1,7)
1,24
9,5 (7,4)
1,34
5,5 (6,9)
0,77
Legenda: ADM = amplitude de movimento; DP = desvio padrão. Fonte: Dados da pesquisa. Quadro 1 - Média (em graus) de ADM, valores máximo e mínimo (DP) para as diferentes articulações n
planos e variabilidade intrassujeito (média de DP dos indivíduos nas duas visitas) Legenda: ADM = amplitude de movimento; DP = desvio padrão. Fonte: Dados da pesquisa. Fisioter Mov. 2013 jan/mar;26(1):95-105
Quadro 2 - Valores de coeficiente de correlação intraclasse para cada variável de desfecho do estudo
Segmento/Variável
ADM
Valor máximo
Valor mínimo
Retropé sagital
0,98
0,93
0,92
Retropé transverso
0,82
0,73
0,90
Retropé frontal
0,79
0,82
0,63
(Continua) Quadro 2 - Valores de coeficiente de correlação intraclasse para cada variável de desfecho do estudo Fisioter Mov. 2013 jan/mar;26(1):95-105 Desenvolvimento de um modelo de pé segmentado para avaliação de indivíduos calçados 101 Quadro 2 - Valores de coeficiente de correlação intraclasse para cada variável de desfecho do estudo
Segmento/Variável
ADM
Valor máximo
Valor mínimo
Antepé frontal
0,65
0,80
0,79
Antepé transverso
0,60
0,72
0,75
Legenda: ADM = amplitude de movimento. Fonte: Dados da pesquisa. Retropé-perna (Conclusão) 40
15
10
5
0
-5
-10
Retropé Sagital
% Fase de apoio
% Fase de apoio
Retropé Frontal
Ângulo (graus)
Ângulo (graus)
30
20
10
0
0
10
20
30
40
50
60
70
80
90
100
0
10
20
30
40
50
60
70
80
90
100 40
15
10
5
0
-5
-10
-10
-15
-20
-25
Retropé Sagital
% Fase de apoio
% Fase de apoio
% Fase de apoio
Retropé Frontal
Retropé Tranverso
Ângulo (graus)
Ângulo (graus)
Ângulo (graus)
30
20
10
0
0
10
20
30
40
50
60
70
80
90
100
0
10
20
30
40
50
60
70
80
90
100
0
10
20
30
40
50
60
70
80
Média
+/- Desvio Padrão
90
100
Gráfico 1 - Deslocamento angular do retropé durante a fase de apoio da marcha de um indivíduo representativo. Dorsiflexão
(+), flexão plantar (-); inversão (+), eversão (-); adução (+), abdução (-)
Fonte: Dados da pesquisa. 40
Retropé Sagital
% Fase de apoio
Ângulo (graus)
30
20
10
0
0
10
20
30
40
50
60
70
80
90
100 Ângulo (graus) -10
-15
-20
-25
% Fase de apoio
% Fase de apoio
Retropé Tranverso
Ângulo (graus)
0
10
20
30
40
50
60
70
80
90
100
0
10
20
30
40
50
60
70
80
Média
+/- Desvio Padrão
90
100
Gráfico 1 - Deslocamento angular do retropé durante a fase de apoio da marcha de um indivíduo representativo. Dorsiflexã
(+), flexão plantar (-); inversão (+), eversão (-); adução (+), abdução (-) Gráfico 1 - Deslocamento angular do retropé durante a fase de apoio da marcha de um indivíduo representativo. Dorsiflexão
(+), flexão plantar (-); inversão (+), eversão (-); adução (+), abdução (-)
Fonte: Dados da pesquisa. Fisioter Mov. 2013 jan/mar;26(1):95-105 Resende RA, Nascimento LR, Silva MCL, Pinheiro AC, Fonseca ST, Kirkwood RN. 102 6
4
2
0
-2
-4
-6
4
0
10
20
30
40
50
60
70
80
90
100
5
6
7
8
9
10
11
0
10
20
30
40
50
60
70
80
90
100
Antepé Frontal
Antepé Transverso
% Fase de apoio
% Fase de apoio
Ângulo (graus)
Ângulo (graus)
Média
+/- Desvio Padrão
Gráfico 2 - Deslocamento angular do antepé durante a fase de apoio da marcha de um indivíduo representativo. Retropé-perna Inversão
(+), eversão (-); adução (+), abdução (-)
Fonte: Dados da pesquisa 6
4
2
0
-2
-4
-6
0
10
20
30
40
50
60
70
80
90
100
Antepé Frontal
% Fase de apoio
Ângulo (graus) Antepé Frontal 4
0
10
20
30
40
50
60
70
80
90
100
5
6
7
8
9
10
11
Antepé Transverso
% Fase de apoio
Ângulo (graus) Antepé Transverso Gráfico 2 - Deslocamento angular do antepé durante a fase de apoio da marcha de um indivíduo representativo. Inversão
(+), eversão (-); adução (+), abdução (-)
Fonte: Dados da pesquisa Fisioter Mov. 2013 jan/mar;26(1):95-105 Discussão confiabilidade teste-reteste de moderada a excelente, o
que demonstra que o método apresentado é confiável
para a avaliação do deslocamento angular dos seg-
mentos do pé durante a fase de apoio da marcha com
indivíduos calçados. Além disso, a variabilidade intras-
sujeito foi reproduzível, apesar do uso de marcadores
de superfície e do movimento da pele. As diferenças
entre sujeitos foram maiores, indicando que o modelo O método apresentado no presente estudo pos-
sibilitou a avaliação do deslocamento angular em
três dimensões dos segmentos do pé com o indiví-
duo calçado por meio de um sistema baseado em
coordenadas de marcadores anatômicos. As variá-
veis de desfecho do presente estudo apresentaram Fisioter Mov. 2013 jan/mar;26(1):95-105 Desenvolvimento de um modelo de pé segmentado para avaliação de indivíduos calçados 103 é sensível o suficiente para demonstrar a variabilidade
entre sujeitos sem nenhuma deformidade aparente. A forma de adaptação dos calçados no presente
estudo fornece um método simples para mensuração
da movimentação dos segmentos do pé com o indi-
víduo calçado, sem a necessidade de fabricação de
calçados específicos para tal propósito. Esse método
é importante, pois foi demonstrado na literatura que
os movimentos do talão do tênis não são idênticos aos
movimentos do calcâneo dentro do tênis (19). Dessa
forma, com o objetivo de investigar possíveis relações
entre o design do tênis e a cinemática dos segmentos
do pé e a ocorrência de lesões, é recomendada a utili-
zação de métodos que incluam aberturas realizadas
no tênis para fixação dos marcadores diretamente na
pele dos indivíduos (19). Embora seja importante a avaliação da confiabilida-
de dos métodos utilizados, uma apropriada análise de
confiabilidade das medidas de cinemática dos segmen-
tos do pé é encontrada em apenas alguns estudos (8). Segundo Bland e Altman (18), esse processo é reco-
mendado especialmente nas ciências que avaliam o
comportamento motor, uma vez que a maioria das
avaliações envolve possíveis erros de mensuração
relacionados à padronização dos procedimentos e
características do examinador. A boa consistência
observada entre as medidas em que se utilizou o pre-
sente modelo indica que os possíveis erros inerentes
a seus procedimentos, como artefatos decorrentes
da movimentação da pele, são sistemáticos, o que
contribui para a adequada quantificação e interpre-
tação das variáveis de desfecho do presente estudo. Alguns modelos multissegmentados para o pé fo-
ram apresentados previamente na literatura (8, 11). Discussão No entanto, esses modelos tiveram como objetivo
avaliar o deslocamento angular dos segmentos do
pé com o indivíduo descalço. Apesar de possibilitar
maior segmentação do pé, avaliar o deslocamento
angular dos segmentos do pé com o indivíduo descal-
ço pode não captar a influência do calçado utilizado
pelo indivíduo; estudos prévios demonstraram que
andar descalço ou calçado pode modificar o padrão
de movimento dos segmentos do pé (20, 21, 22), e
que diferenças entre os tipos de calçados também
podem alterar o padrão cinemático dos membros
inferiores durante a marcha. Dessa forma, ter um
método confiável para a avaliação do deslocamento
angular dos segmentos do pé com o indivíduo calçado
possibilitará maior entendimento da relação entre as
características do calçado e a cinemática dos mem-
bros inferiores durante a marcha. Os clusters de marcadores rígidos utilizados pos-
sibilitaram melhor fixação sobre as estruturas ósseas
definidas para o rastreamento dos segmentos do que
marcadores fixados diretamente sobre a pele, o que
possivelmente minimizou erros relacionados à movi-
mentação da pele. Além disso, a orientação dos clus-
ters permite a adequada visualização dos marcadores
durante toda a fase de apoio da marcha; isso reduz a
necessidade de utilização de filtros para interpolação
das trajetórias desses marcadores após o término da
coleta de dados. Apesar de os clusters terem perma-
necido fixos sobre as estruturas ósseas durante toda
a coleta dos dados, nenhum sujeito avaliado relatou
qualquer tipo de incômodo ou influência sobre a sua
forma de andar em consequência do método utilizado. A realização de aberturas no tênis é um dos méto-
dos possíveis para medir o movimento dos segmentos
do pé dentro do tênis (19). As dimensões das abertu-
ras utilizadas no presente estudo foram similares às
utilizadas por Stacoff et al. (19), que não encontraram
diferenças nas medidas angulares obtidas em decor-
rência da realização das aberturas. Essas aberturas
não podem ser muito pequenas a ponto de não permi-
tirem a movimentação dos clusters de rastreamento
nem muito grandes a ponto de reduzirem a rigidez do
talão do tênis. As adaptações realizadas nos tênis são
de fácil execução e não requerem maior investimento
financeiro além daquele necessário para comprar os
tênis. Além disso, durante toda a realização do estudo,
nenhum indivíduo relatou insegurança, incômodo ou
falta de firmeza para deambular com o tênis e com
os clusters de rastreamento. Fisioter Mov. 2013 jan/mar;26(1):95-105 Discussão É esperado que métodos diferentes de segmenta-
ção do pé forneçam padrões cinemáticos distintos. Assim, torna-se difícil a comparação dos deslocamen-
tos angulares obtidos com outros estudos em conse-
quência de diferentes definições das referências ana-
tômicas, da posição neutra do pé e das articulações
analisadas. Além disso, alguns estudos reportaram os
dados de somente um sujeito (2, 23), o que dificulta
a extrapolação dos padrões cinemáticos obtidos. Ao
contrário de alguns estudos anteriores (24), o pro-
tocolo de aquisição apresentado no presente estudo
não é invasivo, não requer uma estrutura extra de
referência (7, 11, 25-31), posicionamento da articu-
lação subtalar em neutro (32, 33) nem é dependente
da informação fornecida por raios-X para definição da
posição neutra de referência. Essas técnicas são com-
plexas e demandam muito tempo para a realização Fisioter Mov. 2013 jan/mar;26(1):95-105 Resende RA, Nascimento LR, Silva MCL, Pinheiro AC, Fonseca ST, Kirkwood RN. 104 da coleta de dados, além de, em alguns casos, expor
o indivíduo à radiação. No entanto, possíveis altera-
ções do alinhamento estrutural de retropé e antepé
podem influenciar negativamente na definição dos
eixos articulares baseados no protocolo de marca-
dores deste estudo. Além disso, os valores angulares
máximos e mínimos encontrados podem não repre-
sentar os valores exatos, pois as coletas dinâmicas
foram analisadas tendo como base a coleta estática
com os indivíduos descalços, o que, no entanto, não
influencia no padrão de deslocamento angular e nos
valores de ADM total reportados. 5. Nigg BM, Cole GK, Nachbauer W. Effects of arch
height of the foot on angular motion of the lower ex-
tremities in running. J Biomech. 1993;26(8):909-16. doi:10.1016/0021-9290(93)90053-H. 6. Bellchamber TL, Van Den Bogert AJ. Contributions of pro-
ximal and distal moments to axial tibial rotation during
walking and running. J Biomech. 2000;33(11):1397-
403. doi:10.1016/S0021-9290(00)00113-5. 7. Do natelli R, Wooden M, Ekedahl SR, Wilkes JS,
Cooper J, Bush AJ. Relationship between static and
dynamic foot postures in professional baseball
players. J Orthop Sports Phys Ther. 1999;29(6):316-
25. PMid:10370915. Um método simples e confiável para avaliar o
deslocamento angular dos segmentos do pé duran-
te a marcha com o indivíduo calçado foi apresenta-
do no presente estudo. A qualidade do método de-
senvolvido e da análise de confiabilidade realizada
detectou padrões de movimento dos segmentos do
pé próximos aos descritos na literatura. Discussão O nível de
confiabilidade encontrado foi considerado aceitável,
fornecendo, assim, uma base sólida para a avaliação
objetiva da cinemática dos segmentos do pé com in-
divíduos calçados, em ambientes laboratoriais, para
fins de pesquisas científicas e avaliações clínicas da
cinemática da marcha humana. 8. Ca rson MC, Harrington ME, Thompson N, O'Connor JJ,
Theologis TN. Kinematic analysis of a multi-segment
foot model for research and clinical applications: a
repeatability analysis. J Biomech. 2001;34(10):1299-
307. doi:10.1016/S0021-9290(01)00101-4. 9. Resende RA, Kirkwood RN, Figueiredo EM. Cinemática
da marcha de adultos jovens: dados normativos ini-
ciais. Ter Man. 2010;8(39):370-6. 10. Sc ott SH, Winter DA. Talocrural and talocalcaneal
joint kinematics and kinetics during the stance
phase of walking. J Biomech. 1991;24(8):743-52. doi:10.1016/0021-9290(91)90338-N. Referências 11. Le ardini A, Benedetti MG, Catani F, Simoncini L,
Giannini S. An anatomically based protocol for the
description of foot segment kinematics during gait. Clin Biomech (Bristol. Avon.). 1999;14(8):528-36. doi:10.1016/S0268-0033(99)00008-X. 1. Gage JR, Deluca PA, Renshaw TS. Gait analysis: prin-
ciple and applications with emphasis on its use in
cerebral palsy. Instr Course Lect. 1996;45:491-507. PMid:8727765. 2. Kidder SM, Abuzzahab FS, Harris GF, Johson JE. A system for the analysis of foot and ankle kinematics
during gait. IEEE Trans Rehabil Eng. 1996;4(1):25-32. doi:10.1109/86.486054. 2. Kidder SM, Abuzzahab FS, Harris GF, Johson JE. A system for the analysis of foot and ankle kinematics
during gait. IEEE Trans Rehabil Eng. 1996;4(1):25-32. doi:10.1109/86.486054. 12. Je nkyn TR, Anas K, Nichol A. Foot segment kinema-
tics during normal walking using a multisegment
model of the foot and ankle complex. J Biomech Eng. 2009;131(3):034504. doi:10.1115/1.2907750. 3. Radin EL, Yang KH, Riegger C, Kish VL, O'Connor JJ. Relationship between lower limb dynamics and
knee joint pain. J Orthop Res. 1991;9(3):398-405. doi:10.1002/jor.1100090312. 13. Oe ffinger D, Brauch B, Cranfill S, Hisle C, Wynn C,
Hicks R, et al. Comparison of gait with and without
shoes in children. Gait Posture. 1999;9(2):95-100. doi:10.1016/S0966-6362(99)00005-3. 3. Radin EL, Yang KH, Riegger C, Kish VL, O'Connor JJ. Relationship between lower limb dynamics and
knee joint pain. J Orthop Res. 1991;9(3):398-405. doi:10.1002/jor.1100090312. 4. Kirkwood RN, Resende RA, Magalhães CMB, Gomes HA,
Mingoti SA, Sampaio RF. Application of princi-
pal component analysis on gait kinematics in el-
derly women with knee osteoarthritis. Rev Bras
Fisioter. 2011;15(1):52-8. doi:10.1590/S1413-
35552011000100007. 14. Gard SA, Miff SC, Kuo AD. Comparison of kinema-
tic and kinetic methods for computing the vertical
motion of the body center of mass during walking. Hum Mov Sci. 2004;22(6):597-610. doi:10.1016/j. humov.2003.11.002. Fisioter Mov. 2013 jan/mar;26(1):95-105 Desenvolvimento de um modelo de pé segmentado para avaliação de indivíduos calçados 105 15. Winter DA. Biomechanical motor patterns in nor-
mal walking. J Mot Behav. 1983;15(4):302-30. PMid:15151864. 26. Stebbins J, Harrington M, Thompson N, Zavatsky
A, Theologis T. Repeatability of a model for meas-
uring multi-segment foot kinematics in children. Gait Posture. 2006;23(4):401-10. doi:10.1016/j. gaitpost.2005.03.002. 16. Portney LG, Watkins MP. Foundations of clinical re-
search: aplications to practice. 2nd. ed. New Jersey:
Prentice-Hall Health; 2000. p. 61-77. 27. Hunt AE, Smith RM, Torode M. Extrinsic muscle
activity, foot motion and ankle joint moments dur-
ing the stance phase of walking. Foot Ankle Int. 2001;22(1):31-41. PMid:11206820. 17. Referências Walter SD, Eliasziw M, Donner A. Sample size and
optimal designs for reliability studies. Stat Med. 1998;17:101-10. doi:10.1002/(SICI)1097-0258
(19980115)17:1<101::AID-SIM727>3.3.CO;2-5. 28. Kaufman K, Morrow D, Hansen D, Kitaoka HB. Technique for measurement of foot ankle kinematics. Gait and Clinical Movement Analysis Society – GCMAS. 2001; Sacramento, USA. 18. Bland JM, Altman DG. Measurement error and corre-
lation coefficients. BMJ. 1996;313:41-2. doi:10.1136/
bmj.313.7048.41. 29. Moseley L, Smith R, Hunt A, Gant R. Three-
dimensional kinematics of the rearfoot during the
stance phase of walking in normal young adult ma-
les. Clin Biomech. 1996;11(1):39-45. doi:10.1016/
0268-0033(95)00036-4. 19. Stacoff A, Reinschmidt C, Stüssi E. The move-
ment of the hell within a running shoe. Med Sci
Sports Exerc. 1992;24(6):695-701. doi:10.1249/
00005768-199206000-00012. 20. Butler RJ, Davis IS, Hamill J. Interaction of arch
type and footwear on running mechanics. Am J
Sports Med. 2006;34(12):1998-2005. doi:10.1177/
0363546506290401. 30. Simon J, Doederlein L, McIntosh AS, Metaxiotis D,
Bock HG, Wolf SI. The Heidelberg foot measurement
method: development, description and assessment. Gait Posture. 2006;23(4):411–24. doi:10.1016/j. gaitpost.2005.07.003. 21. Hamill J, Bates BT, Holt KG. Timing of lower ex-
tremity joint actions during treadmill run-
ning. Med Sci Sports Exerc. 1992;24(7):807-13. doi:10.1249/00005768-199207000-00011. 31. Theologis TN, Harrington ME, Thompson N,
Benson MK. Dynamic foot movement in children
treated for congenital talipes equinovarus. J Bone
Joint Surg Br. 2003;85(4):572–7. doi:10.1302/
0301-620X.85B4.13696. 22. Morio C, Lake MJ, Gueguen N, Rao G, Baly L. The in-
fluence of footwear on foot motion during walking
and running. J Biomech. 2009;42(13):2081-8. doi:
10.1016/j.jbiomech.2009.06.015. 32. Kepple TM, Stanhope SJ, Lohmann KN, Roman NL. A vid-
eobased technique for measuring ankle-subtalar mo-
tion during stance. J Biomed Eng. 1990;12(4):273–80. doi:10.1016/0141-5425(90)90001-4. 23. Delozier G, Alexander I, Narayanaswamy R. A meth-
od for measurement of integrated foot kinematics. International Symposium on Three-Dimensional
Analysis of Human Movement, Montreal, Canada. 1991:79-82. 33. Woodburn J, Turner DE, Helliwell PS, Barker S. A pre-
liminary study determining the feasibility of electro-
magnetic tracking for kinematics at the ankle joint
complex. Rheumatology (Oxford). 1999;38(12):1260–
8. doi:10.1093/rheumatology/38.12.1260. 33. Woodburn J, Turner DE, Helliwell PS, Barker S. A pre-
liminary study determining the feasibility of electro-
magnetic tracking for kinematics at the ankle joint
complex. Rheumatology (Oxford). 1999;38(12):1260–
8. doi:10.1093/rheumatology/38.12.1260. 24. Chen WP, Tang FT, Ju CW. Stress distribution of the foot
during mid-stance to push-off in barefoot gait: a 3-D fi-
nite element analysis. Clin Biomech. 2001;16(7):614-
20. doi:10.1016/S0268-0033(01)00047-X. Recebido: 15/12/2011
Received: 12/15/2011 25. Rattanaprasert U, Smith R, Sullivan M, Gilleard W. Three-dimensional kinematics of the forefoot, rear-
foot, and leg without the function of tibialis pos-
terior in comparison with normals during stance
phase of walking. Clin Biomech. 1999;14(1):14–23. doi:10.1016/S0268-0033(98)00034-5. Aprovado: 16/07/2012
Approved: 07/16/2012 Fisioter Mov. 2013 jan/mar;26(1):95-105
|
https://openalex.org/W2972159071
|
https://zenodo.org/record/3399540/files/20595__1_233475_LE_337072.pdf
|
Dutch
| null |
Consolidatie-uitsluiting wegens afwijkende bedrijfsactiviteiten
|
MAB
| 1,990
|
cc-by
| 6,907
|
MAB MAB Drs. Ruud G. A. Vergoossen studeerde bedrijfseconomie
aan de Erasmus Universiteit te Rotterdam. Thans is hij als
universitair docent verbonden aan de Faculteit der
Economische Wetenschappen en Econometrie van de
Vrije Universiteit te Amsterdam. Consolidatie-uitsluiting
wegens afwijkende
bedrijfsactiviteiten Drs. Ruud G. A. Vergoossen groepsmaatschappijen met afwijkende bedrijfs
activiteiten niet te worden meegeconsolideerd,
indien consolidatie in strijd zou zijn met het wette
lijk vereiste inzicht.3 Dit is conform de Zevende
EG-richtlijn. In de Verenigde Staten daarentegen
is in 1988 de consolidatie-uitsluiting wegens
branchevreemde activiteiten komen te vervallen. Ook het International Accounting Standards
Committee (IASC) heeft de uitsluitingsclausule
wegens afwijkende bedrijfsactiviteiten onlangs
geschrapt. 1 Inleiding1 De activiteiten van met name concerns vinden
doorgaans plaats in afzonderlijke juridische een
heden. Ter verkrijging van een adequaat inzicht in
de financiële positie en de behaalde resultaten
van een concern is vooral de geconsolideerde
jaarrekening van belang. De vennootschappelijke
jaarrekening van de moedermaatschappij speelt
hierbij een ondergeschikte rol, hoewel formeel
bezien de geconsolideerde jaarrekening slechts
een onderdeel is van de toelichting op deze ven
nootschappelijke jaarrekening. In dit artikel worden de achtergronden van beide
visies omtrent deze uitsluitingsclausule nader
beschouwd en met elkaar vergeleken. Verder
wordt nagegaan in hoeverre in Nederland een
beroep wordt gedaan op de uitzonderingsclau-
sule wegens afwijkende bedrijfsactiviteiten en in
hoeverre dit terecht is geschied. Echter alvorens
dit te doen zal ik in het kort ingaan op de effecten
van consolidatie. Van belang in dit kader is de bepaling van de con
solidatiekring. Met andere woorden, welke maat
schappijen moeten met de jaarrekening van de
moedermaatschappij
worden
meegeconsoli
deerd? In de Verenigde Staten geldt dat in principe alle
deelnemingen waarin de moedermaatschappij
meer dan de helft van de stemrechten heeft
(majority-owned subsidiaries) in de consolidatie
dienen te worden betrokken. In ons land staat bij
de bepaling van de consolidatiekring het groeps-
begrip
centraal. Een
groep
wordt
daarbij
omschreven als een economische eenheid
waarin rechtspersonen en vennootschappen
organisatorisch zijn verbonden. De consolida
tiekring wordt in principe gevormd door de maat
schappij die aan het hoofd van de groep staat en
de groepsmaatschappijen.2 Jaarrekening
Wet- en regelgeving
Onderzoek Jaarrekening
Wet- en regelgeving
Onderzoek Jaarrekening
Wet- en regelgeving
Onderzoek MAB post ’Resultaat uit deelnemingen’ opgenomen
resultaat van deze deelneming uitgesplitst in de
afzonderlijke opbrengsten- en kostenposten. Wanneer een deelneming wordt gewaardeerd
tegen nettovermogenswaarde4 heeft consolidatie
geen invloed op het in de geconsolideerde balans
opgenomen eigen vermogen. Ook het eindcijfer
van de geconsolideerde resultatenrekening blijft
dan ongewijzigd. ving the growing problem of off-balance-sheet
financing. Unconsolidated majority-owned sub
sidiaries have been a significant aspect of that
problem. The growing size and importance of
finance and other unconsolidated majority-
owned subsidiaries and the resulting amounts of
assets, liabilities, revenues, and expenses that
have not been reflected in many consolidated
financial statements have made the matter impor
tant.’ post ’Resultaat uit deelnemingen’ opgenomen
resultaat van deze deelneming uitgesplitst in de
afzonderlijke opbrengsten- en kostenposten. Wanneer een deelneming wordt gewaardeerd
tegen nettovermogenswaarde4 heeft consolidatie
geen invloed op het in de geconsolideerde balans
opgenomen eigen vermogen. Ook het eindcijfer
van de geconsolideerde resultatenrekening blijft
dan ongewijzigd. Het totaal vermogen in de geconsolideerde
balans wordt daarentegen wel beïnvloed. De
balanstellingen nemen namelijk toe met het
bedrag van de schulden van de geconsolideerde
deelneming en met het minderheidsaandeel van
derden indien en voor zover het geen 100%-deel-
neming betreft en in dat geval de integrale
methode van consolidatie wordt toegepast. Aldus is de omvang en samenstelling van de con
solidatiekring mede bepalend voor de gepresen
teerde rentabiliteit, liquiditeit alsmede solvabili
teit. Met het van kracht worden van SFAS 94 is de
zogenaamde ’nonhomogeneity exception’ zoals
geformuleerd in Accounting Research Bulletin
No. 51 (ARB 51) komen te vervallen. In paragraaf
3 van ARB 51 wordt onder meer het volgende ver
meld: ’... For example, separate statements may
be required for a subsidiary which is a bank or an
insurance company and may be preferable for a
finance company where the parent and the other
subsidiaries are engaged in manufacturing ope
rations.’ (cursief RGAV). Met name het cursief
gedrukte gedeelte heeft aanleiding gegeven tot
het oneigenlijk gebruik van de uitsluitingsclausule
wegens branchevreemde activiteiten. Door consolidatie worden niet alleen de verhou
dingen op de geconsolideerde balans beïnvloed. Het al dan niet meeconsolideren van een deelne
ming kan ook invloed hebben op het beeld dat de
geconsolideerde winst- en verliesrekening en
(eventueel) de geconsolideerde staat van her
komst en besteding van middelen tonen. g
International Accounting Standard 27 (IAS 27),
’Consolidated Financial Statements and Ac
counting for Investments in Subsidiaries’, sluit
nauw aan bij SFAS 94. MAB Het IASC is van mening dat
consolidatie-uitsluiting van ’subsidiaries’5 met
afwijkende
bedrijfsactiviteiten
geenszins
is
gerechtvaardigd. Volgens het IASC is het beter
deze ondernemingen gewoon mee te consolide
ren. Wel moet dan additionele informatie worden
verstrekt over de diverse bedrijfsonderdelen
(paragraaf 12, IAS 27). Vermeldenswaardig in dit
kader is dat een verzoek van de Raad voor de
Jaarverslaggeving aan het IASC om consolidatie
van
groepsmaatschappijen
met
afwijkende
bedrijfsactiviteiten te verbieden, indien dat het
inzicht in de financiële positie en de behaalde
resultaten van het concern in gevaar zou brengen,
niet is gehonoreerd.6 2 Effecten van consolidatie De geconsolideerde jaarrekening wordt beïn
vloed door de omvang en samenstelling van de
consolidatiekring. Door het meeconsolideren van
een deelneming wordt de balanspost ’Deelne
mingen’, voor zover betrekking hebbend op deze
deelneming, uitgesplitst in de afzonderlijke bezit
tingen en schulden van de deelneming. Verder
wordt in de winst- en verliesrekening het in de Voor wat betreft de uitzonderingen die op de
gehanteerde consolidatieregels gelden, bestaat
tussen de Nederlandse wetgeving en Ameri
kaanse regelgeving een significant verschil. Vol
gens de Nederlandse wetgeving namelijk dienen 290 juli/augustus 1990 juli/augustus 1990 MAB (Heian & Thies, 1989). Door het niet meeconsoli
deren van de financieringsdochters bleek dat
maar liefst $ 230 miljard vreemd vermogen buiten
de geconsolideerde balans werd gehouden. Con
solidatie van de financieringsdochters zou leiden
tot een stijging van de verhouding vreemd vermo-
gen/totaal vermogen met gemiddeld 12% (van
58% naar 65%). Deze stijging varieerde voor de
diverse bedrijven van 0% tot 80%. Bovendien
werd geconstateerd dat in de periode 1975-1985
een vervijfvoudiging in deze vorm van ’off-
balance-sheet’-financiering had plaatsgevonden. Mohr (1988), die behalve de invloed van consoli
datie van de financieringsdochters op de gepre
senteerde
vermogensverhoudingen
ook
de
karakteristieken van de financieringsdochters
onderzocht, kwam tot soortgelijke conclusies. (Heian & Thies, 1989). Door het niet meeconsoli
deren van de financieringsdochters bleek dat
maar liefst $ 230 miljard vreemd vermogen buiten
de geconsolideerde balans werd gehouden. Con
solidatie van de financieringsdochters zou leiden
tot een stijging van de verhouding vreemd vermo-
gen/totaal vermogen met gemiddeld 12% (van
58% naar 65%). Deze stijging varieerde voor de
diverse bedrijven van 0% tot 80%. Bovendien
werd geconstateerd dat in de periode 1975-1985
een vervijfvoudiging in deze vorm van ’off-
balance-sheet’-financiering had plaatsgevonden. Mohr (1988), die behalve de invloed van consoli
datie van de financieringsdochters op de gepre
senteerde
vermogensverhoudingen
ook
de
karakteristieken van de financieringsdochters
onderzocht, kwam tot soortgelijke conclusies. Interessant in dit kader is ook het onderzoek van
Livnat & Sondhi (1986). Zij geven aan dat het niet
meeconsolideren van de vooral met vreemd ver
mogen gefinancierde financieringsdochters niet
alleen leidt tot een verbetering van de gepresen
teerde financiële ratio’s, maar ook dat de variabili
teit van deze ratio’s vermindert. Hierdoor wordt
de kans kleiner dat afspraken met kredietver
schaffers omtrent de financiële ratio’s worden ge
schonden. afwijkende bedrijfsactiviteiten. Door de Raad
voor de Jaarverslaggeving werd een dergelijke
clausule opgenomen in een ontwerp-Richtlijn
(paragraaf 2.03.303). (p
g
)
Bij de verwerking van de Zevende EG-richtlijn in
de Nederlandse wetgeving is de consolidatiever-
plichting geherformuleerd en vastgelegd in artikel
406 BW 2. Lid 1 luidt als volgt: ’De rechtspersoon
die... aan het hoofd staat van zijn groep, neemt in
de toelichting van zijn jaarrekening een geconso
lideerde jaarrekening op van de eigen financiële
gegevens met die van zijn dochtermaatschap
pijen in de groep en andere groepsmaatschap
pijen’. 3 Regelgeving en praktijk in de
Verenigde Staten In tegenstelling tot de Nederlandse wetgever en
de Raad van de Europese Gemeenschappen is
de Financial Accounting Standards Board (FASB)
van mening dat ’majority-owned subsidiaries’
met afwijkende bedrijfsactiviteiten te allen tijde in
de consolidatie dienen te worden opgenomen. Dit
kan al worden afgeleid uit de titel van het betref
fende Statement (SFAS 94): ’Consolidation of AH
Majority-owned Subsidiaries’ (cursief RGAV). De
reden om een uitzondering wegens afwijkende
bedrijfsactiviteiten niet op te nemen werd ingege
ven door het feit dat in het verleden deze uitzon-
deringsclausule oneigenlijk werd gebruikt en de
vrees dat dit in de toekomst nog verder zou kun
nen toenemen. Dit blijkt onder andere uit para
graaf 22: This Statement is a major step in resol- Uit diverse onderzoeken in de Verenigde Staten is
het oneigenlijk gebruik van de ’nonhomogeneity
exception’ gebleken. Zo werden de jaarrekenin
gen van 140 ondernemingen met niet meegecon
solideerde
financieringsdochters
onderzocht 291 juli/augustus 1990 MAB Echter wanneer de werkzaamheden van
een groepsmaatschappij zodanig afwijken van de
andere activiteiten van de groep dat consolidatie
strijdig zou zijn met het door de geconsolideerde
jaarrekening krachtens artikel 362 lid 1 BW 2 te
verschaffen inzicht, dient de desbetreffende
groepsmaatschappij niet te worden meegecon
solideerd. De jaarrekening van deze groepsmaat
schappij moet dan afzonderlijk in de toelichting
worden opgenomen. Deze uitzondering wegens
afwijkende bedrijfsactiviteiten is vastgelegd in lid
3 van genoemd artikel. De Zevende EG-richtlijn
verplicht de EG-lidstaten onderhavige uitslui-
tingsclausule in de regelgeving op te nemen (arti
kel 14 lid 1).8
' Interessant in dit kader is ook het onderzoek van
Livnat & Sondhi (1986). Zij geven aan dat het niet
meeconsolideren van de vooral met vreemd ver
mogen gefinancierde financieringsdochters niet
alleen leidt tot een verbetering van de gepresen
teerde financiële ratio’s, maar ook dat de variabili
teit van deze ratio’s vermindert. Hierdoor wordt
de kans kleiner dat afspraken met kredietver
schaffers omtrent de financiële ratio’s worden ge
schonden. Door de nieuwe wetsbepalingen zijn de mogelijk
heden om groepsmaatschappijen met afwijkende
bedrijfsactiviteiten buiten de consolidatie te hou
den ingeperkt. Onder de oude wet diende conso
lidatie immers ’reeds’ achterwege te blijven
indien consolidatie niet bijdroeg tot het te ver
schaffen inzicht, terwijl onder de nieuwe wet ’pas’
mag worden afgezien van consolidatie wanneer
consolidatie zou leiden tot strijdigheid met dat in
zicht. MAB geschrapt vanwege het oneigenlijk gebruik dat
van een dergelijke uitzonderingsclausule zou
kunnen worden gemaakt. De Raad van de Euro
pese Gemeenschappen en de Nederlandse wet
gever daarentegen zijn daartoe niet overgegaan. Men kan zich nu afvragen in hoeverre in Neder
land de uitsluitingsgrond wegens afwijkende
bedrijfsactiviteiten oneigenlijk wordt gebruikt. Door het NIvRA (1987) en Van der Tas (1987) is
onderzocht in welke mate een beroep werd
gedaan op onderhavige uitsluitingsclausule. In
deze onderzoeken zijn de jaarrekeningen onder
zocht van beursgenoteerde ondernemingen over
1986 respectievelijk 1983/1985. Uit het onder
zoek van het NIvRA is gebleken dat slechts twee
keer een beroep werd gedaan op de uitzonde
ringsclausule (onderzoekspopulatie: 120). Van
der Tas telde in de jaarrekeningen over 1985 vier
gevallen (onderzoekspopulatie: 125). 12 Hieruit
kan men afleiden dat de uitsluitingsclausule
wegens branchevreemde activiteiten in Neder
land slechts sporadisch wordt gebruikt. neer de activiteiten van groepsmaatschappijen
dusdanig afwijken van de andere activiteiten van
de groep dat het consolideren van bedoelde
groepsmaatschappijen zou leiden tot een niet
getrouwe weergave van groepsvermogen en
-resultaat. Wel heeft de minister van Justitie in zijn
Memorie van Antwoord een voorbeeld genoemd,
te weten een bank of verzekeringsmaatschappij
die groepsmaatschappij is in een groep met
andere hoofdwerkzaamheid9 (vergelijk het in de
vorige paragraaf aangehaalde gedeelte uit ARB
51). Door de Raad voor de Jaarverslaggeving wordt
dit in een zogenaamde extra publikatie (het
betreft hier geen officiële richtlijn) nader gemoti
veerd. De Raad meldt namelijk het volgende:10 ’In
vele gevallen zullen afwijkende bedrijfsactivitei
ten bij consolidatie niet verstorend werken op het
beeld dat de geconsolideerde jaarrekening
oproept.11 Er zijn echter combinaties van activi
teiten binnen één concern die moeilijk op
dezelfde basis verantwoord kunnen worden. Te
denken valt daarbij bijvoorbeeld aan een indus
triële onderneming met een bank- of verzeke
ringsbedrijf. In de winst- en verliesrekening zijn
begrippen als omzet en bedrijfsresultaat nauwe
lijks vergelijkbaar en leiden deze voorts tot weinig
zeggende ratio’s. Met betrekking tot de balans
zijn de financieringsverhoudingen voor beide
activiteiten veelal sterk uiteenlopend, zodat ook
daar consolidatie niet tot een goed inzicht leidt.’
Gaarne sluit ik mij bij de mening van de Raad aan. Ik moet daarbij wel aantekenen dat tussen
groepsmaatschappijen
sterk
uiteenlopende
financieringsverhoudingen sec geen rol mogen
spelen bij de bepaling van de consolidatiekring. MAB Zowel uit het Memorie van Antwoord als uit de
mededeling van de Raad voor de Jaarverslagge
ving kan men afleiden dat de uitzonderingsclau-
sule wegens verschil in werkzaamheden zeer
restrictief moet worden uitgelegd en zoals uit de
vergelijking van de nieuwe met de oude wet blijkt,
restrictiever dan in het verleden. Om na te gaan in hoeverre het gebruik van de uit
zonderingsgrond al dan niet gerechtvaardigd is,
heb ik de jaarrekeningen over 1988 onderzocht
van een honderdtal ondernemingen.13 De volgende vijf ondernemingen hebben in de
jaarrekening over 1988 één of meer meerder-
heidsdeelnemingen
niet
meegeconsolideerd
vanwege van de groep afwijkende bedrijfsactivi
teiten: - Vendex International N.V., - Naamloze Vennootschap DSM, - N.V. Koninklijke Nederlandse Vliegtuigenfa-
briek Fokker, - DAF N.V. en Zowel uit het Memorie van Antwoord als uit de
mededeling van de Raad voor de Jaarverslagge
ving kan men afleiden dat de uitzonderingsclau-
sule wegens verschil in werkzaamheden zeer
restrictief moet worden uitgelegd en zoals uit de
vergelijking van de nieuwe met de oude wet blijkt,
restrictiever dan in het verleden. - Koninklijke Ahold N.V. Deze vijf gevallen worden in het navolgende
besproken. Het is van belang zich te realiseren
dat deze jaarrekeningen onder de oude wettelijke
bepalingen inzake de geconsolideerde jaarreke
ning zijn opgesteld en uitgebracht. Uit geen van
de jaarrekeningen kon worden afgeleid dat bij de
vaststelling van de consolidatiekring werd uitge
gaan van de nieuwe wetsbepalingen. Maar zoals
we hiervoor reeds hebben gezien, leidt de nieuwe
wettelijke regeling tot een inperking van de moge 4 Regelgeving in Nederland Vóór de aanpassing van de Nederlandse wetge
ving aan de Zevende EG-richtlijn was de consoli-
datieverplichting vastgelegd in het ’oude’ artikel
379 lid 6 BW 2. Hierin stond vermeld dat de finan
ciële gegevens omtrent dochtermaatschappijen,
al dan niet te zamen met die van de rechtsper
soon (de moedermaatschappij) en van andere
groepsmaatschappijen, opgenomen dienden te
worden in een groepsjaarrekening die volgens de
methode van consolidatie was samengesteld,7
tenzij het wettelijk vereiste inzicht meer werd
gediend door het buiten de consolidatie laten van
de desbetreffende rechtspersonen en vennoot
schappen. Op grond van de laatste zinsnede was
het mogelijk maatschappijen niet mee te consoli
deren, onder andere wegens branchevreemde
werkzaamheden. Aldus bevatte de oude wet
impliciet een
uitzonderingsclausule
wegens Overigens zij vermeld dat de nieuwe wettelijke
bepalingen inzake de geconsolideerde jaarreke
ning dienen te worden toegepast op boekjaren
die na 31 december 1989 aanvangen. De nieuwe
wet mocht echter geheel of ten dele reeds eerder
worden toegepast, mits daar in de toelichting
melding van werd gemaakt. Hoewel in de huidige wet expliciet een uitzonde
ringsclausule wegens afwijkende bedrijfsactivi
teiten staat verwoord, geeft de wet niet aan wan 292 juli/augustus 1990 MAB nadere motivering van de Raad voor de Jaarvers-
laggeving blijkt, is de handelwijze van Vendex en
DSM ook onder de nieuwe wet gerechtvaardigd. Hierbij dient wel te worden aangetekend dat bin
nen de huidige wetgeving in bepaalde gevallen
een bank of verzekeringsmaatschappij, onder
deel uitmakend van een industrieel of handels
concern, toch moet worden meegeconsolideerd. Het meeconsolideren van bijvoorbeeld een kleine
bank in een industrieel concern hoeft immers niet
strijdig te zijn met het door middel van de gecon
solideerde jaarrekening te verschaffen inzicht (in
welk geval consolidatie verplicht is, behoudens
eventueel verwaarloosbaar belang), terwijl dat
inzicht wel meer zou zijn gediend door het buiten
de consolidatie houden van deze bank. lijkheid om groepsmaatschappijen wegens bran
chevreemde activiteiten buiten de consolidatie te
houden. Met andere woorden, indien onder het
oude wettelijke regime ten onrechte een beroep
wordt gedaan op de uitzonderingsclausule
wegens afwijkende bedrijfsactiviteiten dan geldt
dit onder de nieuwe wet a fortiori. Vendex International N. V. en Naamloze Vennoot
schap DSM Vendex International N. V. en Naamloze Vennoot
schap DSM In de geconsolideerde jaarrekening van Vendex
worden in principe alle groepsmaatschappijen
meegeconsolideerd. Staal Bankiers N.V., een
algemene bank waarin Vendex een 79%-belang
heeft, vormt hierop een uitzondering. In het jaar
verslag 1988/89 zegt Vendex hierover het vol
gende: ’De bedrijfsactiviteiten van de meerder-
heidsdeelneming Staal Bankiers N.V. wijken af
van die van het Concern in zijn geheel, waardoor
een beter inzicht in vermogen en resultaat wordt
bereikt met een afzonderlijke verslaggeving.’ De
(geconsolideerde) jaarrekening van Staal Ban
kiers heeft Vendex dan ook separaat opgenomen
als bijlage bij de toelichting. N.V. Koninklijke Nederlandse Vliegtuigenfabriek
Fokkeren DAFN.V. N.V. Koninklijke Nederlandse Vliegtuigenfabriek
Fokkeren DAFN.V. Zowel door Fokker als door DAF worden alle
meerderheidsdeelnemingen in de consolidatie
betrokken, tenzij een beter inzicht wordt verkre
gen door een afzonderlijke verslaggeving. Dit
laatste doet zich volgens Fokker voor bij Aircraft
Financing &Trading B.V., een 100%-deelneming. Fokker meldt daarover in het jaarverslag 1988 het
volgende: ’Gelet op de aard van de onderneming
Aircraft Financing & Trading B.V., te weten finan
ciering en verhuur van vliegtuigen, is de jaarreke
ning van deze onderneming wel geconsolideerd
met die van haar 100%-deelnemingen, doch bui
ten de consolidatie van de vennootschap gehou
den.’ De (geconsolideerde) jaarrekening van
Aircraft Financing & Trading is separaat opge
nomen. Bij DSM doet zich een vergelijkbare situatie voor. DSM heeft namelijk een 100%-deelneming in een
verzekeringsmaatschappij. 5 Praktijk in Nederland Zoals uit het voorgaande blijkt, hebben de FASB
en het IASC de ’nonhomogeneity exception’ 293 juli/augustus 1990 MAB regeling voor discussie vatbaar. Consolidatie zou
naar mijn mening in ieder geval niet strijdig zijn
met het wettelijk vereiste inzicht onder de nieuwe
wetsbepalingen. Voor beide ondernemingen
geldt dat zij ondersteunend zijn voor de verkoop
van de Fokker- respectievelijk DAF-produkten. Door middel van afzetfinanciering, waaronder
leasing en verhuur, wordt geprobeerd de verkoop
te bevorderen. Vanuit die optiek kan men de acti
viteiten van Aircraft Financing & Trading en DAF
Finance Company tot de normale activiteiten van
het Fokker- respectievelijk DAF-concern re
kenen. heeft Ahold een zogenaamde samengevoegde
balans en resultatenrekening van de onroerend
goedmaatschappijen opgenomen. Verder heeft
er (en passant) een eenmalige herwaardering
plaatsgevonden van het in deze maatschappijen
ondergebrachte onroerend goed. Dit alles naar aanleiding van de verwachting dat
Ahold in toenemende mate onroerend goed in
eigendom zal krijgen dat niet in gebruik is bij
groepsmaatschappijen. ’De daaruit voortvloei
ende werkzaamheden verschillen in belangrijke
mate van de werkzaamheden die verband hou
den met onze hoofdactiviteit: de detailhandel in
levensmiddelen en andere consumentenproduk-
ten. ... Teneinde een beter inzicht te geven in de
omvang en de samenstelling van het vermogen
en de behaalde resultaten van de hoofdactiviteit
enerzijds en de onroerend goed activiteiten
anderzijds worden deze laatste ... niet meer
geconsolideerd met de hoofdactiviteit’, aldus
Ahold in het jaarverslag. Ahold beroept zich aldus
op de (verwachte) afwijkende bedrijfsactiviteiten. Consolidatie van Aircraft Financing & Trading zou
bij Fokker slechts een beperkte invloed hebben
op de gerapporteerde solvabiliteit; de solvabili-
teitsratio15 zou dalen van ruim 55% naar onge
veer 53%. Dit is een gevolg van de relatief
beperkte omvang van het bedrijf binnen het
Fokker-concern.16 Bij DAF daarentegen zou het meeconsolideren
van DAF Finance Company tot een substantiële
verlaging van de gepresenteerde solvabiliteit lei
den; de solvabiliteitsratio zou zakken van ruim
34% naar circa 23%. Interessant is een vergelij
king met de Amerikaanse auto-industrie. Tot het
van kracht worden van SFAS 94 was het in de
Amerikaanse auto-industrie namelijk gebruikelijk
om de financieringsactiviteiten buiten de balans
te houden door het niet meeconsolideren van de
financieringsdochters. Zo werd door General
Motors in het jaarverslag over 1982 een ’debt/
equity’-ratio gerapporteerd van ongeveer 1,3;
Ford Motor Co. toonde een verhoudingscijfer van
2,6. Zouden de financieringsmaatschappijen
daarentegen worden meegeconsolideerd, dan
zou deze ratio 3,4 respectievelijk 4,9 bedragen;
een stijging van 170% respectievelijk 89% (Mohr,
1988). Uit de geconsolideerde balans van DAF
kan een ’debt/equity’-ratio worden afgeleid van
2,0. MAB ’NV Pensioenverze
keringsmaatschappij DSM wordt niet in de con
solidatie
betrokken
vanwege
het
branche
vreemde karakter van deze vennootschap’, aldus
DSM in het jaarverslag over 1988. De jaarrekening
van de betreffende verzekeringsmaatschappij is
echter niet opgenomen in het gepubliceerde
DSM-jaarvers lag. De handelwijze van Vendex en DSM om Staal
Bankiers respectievelijk Pensioenverzekerings
maatschappij DSM niet mee te consolideren is
onder de oude wet zonder meer verdedigbaar. Door de sterk afwijkende opbouw van de resulta
ten en de sterk afwijkende aard van de activa en
passiva van deze ondernemingen, wordt het door
Vendex en DSM door middel van de geconsoli
deerde jaarrekening te verschaffen inzicht meer
gediend met het niet meeconsolideren van Staal
Bankiers respectievelijk Pensioenverzekerings
maatschappij DSM. Ook DAF houdt een soortgelijke deelneming bui
ten de consolidatie. Het betreft de 100%-deelne-
ming DAF Finance Company N.V. ’De dochter
maatschappij DAF Finance Company N.V. is niet
in de geconsolideerde jaarrekening van DAF
B.V.14 opgenomen vanwege de van de rest van de
groep afwijkende aard van de activiteiten’, aldus
DAF. De (geconsolideerde) balans en resultaten
rekening van DAF Finance Company zijn opgeno
men in de toelichting. Het niet meeconsolideren van Aircraft Financing
& Trading en DAF Finance Company door Fokker
respectievelijk DAF is onder de oude wettelijke pp j
Zoals uit het in de vorige paragraaf aangehaalde
voorbeeld van de minister van Justitie en de 294 juli/augustus 1990 MAB Consolidatie van DAF Finance Company zou
evenwel leiden tot een stijging met 70% tot 3,4. Naar mijn mening moeten de activiteiten van de
onroerend goedpoot evenwel tot de normale acti
viteiten van het Ahold-concern worden gerekend. Het betreft hier immers met name winkelpanden
die voor het merendeel worden verhuurd aan
groepsmaatschappijen. Aldus zijn de activiteiten
van de onroerend goedbedrijven (op korte termijn
althans) onmisbaar voor het uitvoeren van de
hoofdactiviteiten. Bovendien bestaat er beleids
matig een nauwe verwevenheid tussen de detail-
handelspoot en de onroerend goedpoot, bijvoor
beeld met betrekking tot de keuze van de vesti
gingsplaatsen.17 Ook wanneer het in de onroerend goedmaat
schappijen ondergebrachte onroerend goed niet
(meer) ten dienste zou staan van de andere
groepsmaatschappijen, zou mijns inziens (zeker
onder de nieuwe wetsbepalingen) tot consolida
tie van de onroerend goedbedrijven moeten wor
den besloten. De diversiteit aan activiteiten bin
nen de groep zou in dat geval niet van dien aard
zijn dat een beroep op de uitzonderingsclausule
wegens afwijkende bedrijsactiviteiten, zou zijn
gerechtvaardigd (vergelijk de eerste zin van de in
de vorige paragraaf aangehaalde passage uit de juli/augustus 1990 6 Evaluatie In de door mij onderzochte jaarverslagen over
1988 werd vijf maal een beroep op de uitsluitings-
clausule wegens afwijkende bedrijfsactiviteiten
geconstateerd. Naar mijn mening was deze
gedragslijn onder de oude wetsbepalingen in drie
gevallen op zijn minst voor discussie vatbaar, ter
wijl onder de huidige wettelijke regeling van een
beroep op de uitzonderingsclausule zou moeten
worden afgezien. Het verschaffen van inzicht in de financiële positie
en de behaalde resultaten van de verschillende
activiteiten van het Ahold-concern is geen reden
om tot deconsolidatie van de onroerend goed
sector over te gaan. Het ware beter geweest om
naast de geconsolideerde jaarrekening (inclusief
de onroerend
goedsector)
gediversificeerde
informatie te verschaffen over de diverse bedrijfs
onderdelen. In het specifieke geval van Ahold kan
men evenwel vraagtekens zetten bij het nut van
het separaat tonen van de resultaten van de
onroerend goedsector; deze worden immers
sterk beïnvloed door de prijsstelling van ’inter-
company’-transacties en zijn daardoor min of
meer kunstmatig van aard. Overigens moet wor
den opgemerkt dat de mate waarin en de wijze
waarop zowel geaggregeerde als gesegmen
teerde informatie wordt verstrekt, in principe los
dient te staan van de wijze waarop een onderne
ming is opgebouwd; het maakt daarbij niet uit of
de diverse activiteiten zijn ondergebracht in
afzonderlijke juridische eenheden of dat alle acti
viteiten zijn ondergebracht in één juridische een
heid (met diverse afdelingen). Van toestanden zoals die zich in de Verenigde
Staten voordeden, is in Nederland geen sprake. Toch dient te worden voorkomen dat de wijze
waarop Ahold, DAF en Fokker de uitzonderings
clausule interpreteren een precedent schept voor
andere bedrijven, zeker nu de uitsluitingsclausule
in de nieuwe wet restrictiever is geformuleerd. Het eventueel laten vervallen van de uitzonde
ringsclausule a la IAS 27 of SFAS 94 is naar mijn
mening niet wenselijk. Nog afgezien van de vraag
of dat binnen het kader van de Zevende EG-richt-
lijn wel mogelijk is. In Nederland geeft Marseille (1989, pagina 74)
aan wel voor een dergelijke regeling te voelen
indien de regelgeving met betrekking tot de door
ondernemingen te verstrekken gesegmenteerde
informatie wordt uitgebreid. Naar mijn mening is
deze oplossing niet bevredigend. MAB mededeling van de Raad voor de Jaarverslagge-
ving). onroerend goedsector, gecombineerd met de
herwaardering van het onroerend goed, had
onder andere tot gevolg dat de getoonde solvabi-
liteitsratio in 1988 niet daalde van 32% naar 24%,
maar van 32% naar 28%. De daling van de sedert
1982 vrij stabiele solvabiliteitsratio (ongeveer
32%) werd aldus met ongeveer de helft terugge
bracht. De verslechtering van de feitelijke vermo
genspositie was met name een gevolg van het
hoge investeringsniveau ( ruim ƒ 1 miljard) in
1988. Zo werd in de Verenigde Staten de FNS-
supermarktketen overgenomen. Het door middel van een samengevoegde balans
en resultatenrekening verschaffen van inzicht in
de financiële positie en de resultaten van de
onroerend goedsector afzonderlijk gaat ten koste
van het inzicht in de financiële positie en de
behaalde resultaten van het Ahold-concern als
één geheel. Vermeldenswaardig in dit kader is dat
in 1982 door Ahold nog het besluit werd genomen
alle deelnemingen in onroerend goedmaatschap-
pijen te consolideren juist ter verbetering van
genoemd inzicht. De beslissing van Ahold om de
onroerend goedbedrijven te deconsolideren is
des te opmerkelijker, indien men beziet dat onder
de nieuwe wet de uitzonderingsclausule wegens
branchevreemde activiteiten restrictiever moet
worden toegepast. Bovendien wordt door Ahold
notering op een Amerikaanse beurs nage
streefd.18 De jaarstukken van Ahold zullen dan in
principe aan de Amerikaanse verslaggevingsre-
gels moeten voldoen. Koninklijke Ahold N. V. Koninklijke Ahold N. V. In de groepsjaarrekening van Ahold over 1988 zijn
in tegenstelling tot voorgaande jaren de deelne
mingen in onroerend goedmaatschappijen niet
meer meegeconsolideerd. Daarvoor in de plaats 295 juli/augustus 1990 juli/augustus 1990 6 Evaluatie Ik ben het eens
met Timmers (1989, pagina 19) als hij zegt: ’Seg
mentatie kan niet als een echte oplossing
beschouwd worden omdat het ’’kwaad” dan al
geschied is, immers in de primaire balans en
winst- en verliesrekening zijn de per definitie niet
bij elkaar op te tellen activiteiten al bij elkaar Het niet meeconsolideren door Ahold van de
vooral met vreemd vermogen gefinancierde 296 Koninklijke Nederlandsche Petroleum Maatschappij,
Den Haag
Koninklijke Nederlandsche Springstoffenfabrieken N.V.,
Bussum
Grand Hotel Krasnapolsky N.V., Amsterdam
Koninklijke Landré & Glinderman N.V., Diemen
Macintosh N.V., Stein
N.V. Medicopharma, Zaandam
Nagron Nationaal Grondbezit N.V., Den Haag
NBM-Amstelland N.V., Den Haag
N.V. Nederlandsche Apparatenfabriek Nedap, Groenlo
Koninklijke Nedlloyd Groep N.V., Rotterdam
NKF Holding N.V., Delft
Norit N.V., Amersfoort
N.V. Verenigde Bedrijven Nutricia, Zoetermeer
Koninklijke Nijverdal-Ten Cate N.V., Almelo
Océ-van der Grinten N.V., Venlo3
Van Ommeren Ceteco N.V., Rotterdam
Oldelft Groep N.V., Delft
OTRA N.V., Amstelveen
Pakhoed Holding N.V., Rotterdam
Palthe N.V., Almelo opgeteld. Dit moet dan gerepareerd worden door
middel van de toelichting.’ Het ene gat (potentieel
misbruik) wordt als het ware gestopt met het
andere (in bepaalde gevallen geen adequaat
inzicht in de financiële positie en de behaalde
resultaten van het concern als economische een
heid). Naar mijn mening kan de uitsluitingsclausule
zoals die in de huidige wet staat geformuleerd wel
verder worden aangescherpt. Bijvoorbeeld door
in de Richtlijnen voor de jaarverslaggeving expli
ciet aan te geven in welke situaties sprake is van
afwijkende bedrijfsactiviteiten die het niet mee
consolideren van een groepsmaatschappij recht
vaardigen. Hetgeen de Raad voor de Jaarver
slaggeving over dit onderwerp heeft gemeld in de
hiervoor genoemde extra publikatie kan daartoe
als aanzet dienen. Door een nadere concretise
ring in de Richtlijnen van artikel 406 lid 3 BW 2 kan
de ’manoeuvreerruimte’, voor zowel de opstellers
als de controleurs van jaarrekeningen, worden
beperkt zonder dat dit ten koste gaat van het
inzicht dat een geconsolideerde jaarrekening
dient te verschaffen. Bijlage: Lijst van ondernemingen
betrokken bij het onderzoek
Aalberts Industries N.V., Venlo
ACF Holding N.V., Maarssen
Koninklijke Ahold N.V., Zaandam
Ahrend Groep N.V., Amsterdam
Akzo N.V., Arnhem
Atag Holding N.V., Ulft
BAM Holding N.V., Bunnik
Batenburg Beheer N.V., Rotterdam
Beers N.V., Den Haag
N.V. Koninklijke Nederlandsche Machinefabriek v/h
E. H. Begemann, Helmond
Maatschappij van Berkel’s Patent N.V., Rijswijk
Blydenstein-Willink N.V., Enschede
De Boer Winkelbedrijven N.V., Beilen
N.V. Koninklijke Distilleerderijen Erven Lucas Bols,
Nieuw-Vennep
Koninklijke Borsumij Wehry N.V., Den Haag
Koninklijke Boskalis Westminster N.V., Papendrecht
Bührmann-Tetterode N.V., Amsterdam
Center Paros N.V., Rotterdam
N.V. Centrale Suiker Maatschappij, Diemen3
Cindu-Key & Kramer N.V., Uithoorn
DAF N.V., Eindhoven Bijlage: Lijst van ondernemingen
betrokken bij het onderzoek
Aalberts Industries N.V., Venlo
ACF Holding N.V., Maarssen
Koninklijke Ahold N.V., Zaandam
Ahrend Groep N.V., Amsterdam
Akzo N.V., Arnhem
Atag Holding N.V., Ulft
BAM Holding N.V., Bunnik
Batenburg Beheer N.V., Rotterdam
Beers N.V., Den Haag
N.V. Tapijtfabriek H. Desseaux N.V., Ossb
Naamloze Vennootschap DSM, Heerlen
Elsevier N.V., Amsterdam
Eriks Holding N.V., Alkmaar
Flexovit International N.V., Eibergen
N.V. Koninklijke Nederlandse Vliegtuigenfabriek
Fokker, Amsterdam
Frans Maas Beheer N.V., Venlo
Furness N.V., Rotterdam
Gamma Holding N.V., Helmond
Getronics N.V., Amsterdam
Van der Giessen-de Noord N.V., Krimpen aan den IJssel
Koninklijke Gist-brocades N.V., Delft
Goudsmit N.V., De Bilt
Grasso’s Koninklijke Machinefabrieken N.V.,
Den Bosch
Grolsche Bierbrouwerij N.V., Enschede
N.V. GTI Holding, Nieuwegein
Hagemeyer N.V., Naarden
HCS Technology N.V., Apeldoorn
Heineken N.V., Amsterdam
N.V. W. A. Hoek’s Machine- en Zuurstoffabriek, Schiedam
Holland Sea Search Holding N.V., Den Haag
Hollandia Kloos Holding N.V., Krimpen aan den IJssel
Koninklijke Nederlandsche Hoogovens en Staalfabrieken
N.V., IJmuiden
Hunter Douglas N.V., Rotterdam
Ingenieurs Bureau Bouwnijverheid-Kondor Groep N.V.,
Leiden
IHC Caland N.V., Schiedam
N.V. Industrieele Maatschappij, Den Haag
Internatio-Müller N.V., Rotterdam
N.V. Koninklijke Bijenkorf Beheer KBB, Amsterdam0
Koninklijke Luchtvaart Maatschappij N.V., Amstelveen13
Koninklijke Nederlandse Papierfabrieken N.V., Naarden
N.V. Koninklijke Nederlandsche Petroleum Maatschappij,
Den Haag
Koninklijke Nederlandsche Springstoffenfabrieken N.V.,
Bussum
Grand Hotel Krasnapolsky N.V., Amsterdam
Koninklijke Landré & Glinderman N.V., Diemen
Macintosh N.V., Stein
N.V. Medicopharma, Zaandam
Nagron Nationaal Grondbezit N.V., Den Haag
NBM-Amstelland N.V., Den Haag
N.V. Nederlandsche Apparatenfabriek Nedap, Groenlo
Koninklijke Nedlloyd Groep N.V., Rotterdam
NKF Holding N.V., Delft
Norit N.V., Amersfoort
N.V. Verenigde Bedrijven Nutricia, Zoetermeer
Koninklijke Nijverdal-Ten Cate N.V., Almelo
Océ-van der Grinten N.V., Venlo3
Van Ommeren Ceteco N.V., Rotterdam
Oldelft Groep N.V., Delft
OTRA N.V., Amstelveen
Pakhoed Holding N.V., Rotterdam
Palthe N.V., Almelo Tapijtfabriek H. Desseaux N.V., Ossb
Naamloze Vennootschap DSM, Heerlen
Elsevier N.V., Amsterdam
Eriks Holding N.V., Alkmaar
Flexovit International N.V., Eibergen
N.V. Koninklijke Nederlandse Vliegtuigenfabriek
Fokker, Amsterdam
Frans Maas Beheer N.V., Venlo
Furness N.V., Rotterdam
Gamma Holding N.V., Helmond
Getronics N.V., Amsterdam
Van der Giessen-de Noord N.V., Krimpen aan den IJssel
Koninklijke Gist-brocades N.V., Delft
Goudsmit N.V., De Bilt
Grasso’s Koninklijke Machinefabrieken N.V.,
Den Bosch
Grolsche Bierbrouwerij N.V., Enschede
N.V. GTI Holding, Nieuwegein
Hagemeyer N.V., Naarden
HCS Technology N.V., Apeldoorn
Heineken N.V., Amsterdam
N.V. W. A. Hoek’s Machine- en Zuurstoffabriek, Schiedam
Holland Sea Search Holding N.V., Den Haag
Hollandia Kloos Holding N.V., Krimpen aan den IJssel
Koninklijke Nederlandsche Hoogovens en Staalfabrieken
N.V., IJmuiden
Hunter Douglas N.V., Rotterdam
Ingenieurs Bureau Bouwnijverheid-Kondor Groep N.V.,
Leiden
IHC Caland N.V., Schiedam
N.V. Industrieele Maatschappij, Den Haag
Internatio-Müller N.V., Rotterdam
N.V. Koninklijke Bijenkorf Beheer KBB, Amsterdam0
Koninklijke Luchtvaart Maatschappij N.V., Amstelveen13
Koninklijke Nederlandse Papierfabrieken N.V., Naarden
N.V. Noten 1 De MAB-redactie dank ik voor het waardevolle commentaar
op de concept-versie van dit artikel. 1 De MAB-redactie dank ik voor het waardevolle commentaar
op de concept-versie van dit artikel. 2 In dit blad is eerder over deze materie geschreven, bijvoorbeeld door Timmers (1989). 3 Deze uitzondering heeft een verplichtend karakter. Andere
(facultatieve) uitzonderingen zijn: (facultatieve) uitzonderingen zijn: - groepsmaatschappijen waarvan het gezamenlijk belang van te
verwaarlozen betekenis is; a Het betreft hier een onderneming met een gebroken
boekjaar; onderzocht is de jaarrekening over 1987/88. - groepsmaatschappijen waarvan de benodigde gegevens
slechts tegen onevenredige kosten of met grote vertraging te
verkrijgen ofte ramen zijn; 5 Het betreft hier een onderneming met een gebroken
boekjaar; onderzocht is de jaarrekening over 1988/89. 5 Het betreft hier een onderneming met een gebroken
boekjaar; onderzocht is de jaarrekening over 1988/89. - groepsmaatschappijen waarin het belang slechts gehouden
wordt ter vervreemding; N.V. Philips’ Gloeilampenfabrieken, Eindhoven
Polynorm N.V., Bunschoten
Reesink N.V., Zutphen
Riva N.V., Hoofddorp
N.V. Rubber Cultuur Maatschappij ’Amsterdam’,
Amsterdam
Samas-Groep N.V., Utrecht13
Schuitema N.V., Amersfoort
Smit Internationale N.V., Rotterdam
N.V. Koninklijke Sphinx, Maastricht5
Verenigde machinefabrieken Stork N.V., Naarden
N.V. Holdingmaatschappij De Telegraaf, Amsterdam
Textielgroep Twenthe N.V., Almelo
Tulip Computers N.V., Den Bosch
N.V. Twenfsche Kabel Holding, Haaksbergen
Ubbink N.V., Doesburg
Unigro N.V., Houten5
UnileverN.V., Rotterdam
Union N.V., Nieuwleusen
Vendex International N.V., Amsterdam5
N.V. Verto, Rotterdam
N.V. Verenigd Bezit VNU, Haarlem
Koninklijke Volker Stevin N.V., Rotterdam
Volmac Software Groep N.V., Utrecht
Vredestein N.V., Velp
VRG-Groep N.V., Diemen
WegenerTijl N.V., Apeldoorn
Koninklijke Wessanen N.V., Amstelveen
Wolters Kluwer N.V., Amsterdam N.V. Philips’ Gloeilampenfabrieken, Eindhoven
Polynorm N.V., Bunschoten
Reesink N.V., Zutphen
Riva N.V., Hoofddorp
N.V. Rubber Cultuur Maatschappij ’Amsterdam’,
Amsterdam
Samas-Groep N.V., Utrecht13
Schuitema N.V., Amersfoort
Smit Internationale N.V., Rotterdam
N.V. Koninklijke Sphinx, Maastricht5
Verenigde machinefabrieken Stork N.V., Naarden
N.V. Holdingmaatschappij De Telegraaf, Amsterdam
Textielgroep Twenthe N.V., Almelo
Tulip Computers N.V., Den Bosch
N.V. Twenfsche Kabel Holding, Haaksbergen
Ubbink N.V., Doesburg
Unigro N.V., Houten5
UnileverN.V., Rotterdam
Union N.V., Nieuwleusen
Vendex International N.V., Amsterdam5
N.V. Verto, Rotterdam
N.V. Verenigd Bezit VNU, Haarlem
Koninklijke Volker Stevin N.V., Rotterdam
Volmac Software Groep N.V., Utrecht
Vredestein N.V., Velp
VRG-Groep N.V., Diemen
WegenerTijl N.V., Apeldoorn
Koninklijke Wessanen N.V., Amstelveen
Wolters Kluwer N.V., Amsterdam Marseille H., Consolidatie en groepsregelingen, in: Beekman H. (red.), Ontwikkelingen in de financiële verslaggeving:
grondslagen en praktijk, Kluwer, Deventer, 1989. Mohr R. M., Unconsolidated Finance Subsidiaries:
Characteristics and Debt/Equity Effects, Accounting Horizons,
maart 1988. Nederlands Instituut van Registeraccountants, Onderzoek
Jaarverslaggeving 1984, NIvRA-geschrift 39, Kluwer,
Deventer, 1985. Nederlands Instituut van Registeraccountants, Onderzoek
Jaarverslaggeving 1986, NIvRA-geschrift 42, Kluwer,
Deventer, 1987. Raad voor de Jaarverslaggeving, Richtlijnen voorde
jaarverslaggeving, Kluwer, Deventer, bijgewerkt tot en met
augustus 1989. Van der Tas L. G., Groepsverslaggeving, in: Dijksma J. (red.),
Jaarin - jaaruit2, Wolters-Noordhoff, Groningen, 1987. Jaarin - jaaruit2, Wolters-Noordhoff, Groningen, 1987 Timmers M. P., Nieuwe wettelijke bepalingen voor de
geconsolideerde jaarrekening, Maandblad voor Accountancy
en Bedrijfseconomie, januari/februari 1989. Timmers M. P., Nieuwe wettelijke bepalingen voor de
geconsolideerde jaarrekening, Maandblad voor Accountancy
en Bedrijfseconomie, januari/februari 1989. Timmers M. P., Nieuwe wettelijke bepalingen voor de
geconsolideerde jaarrekening, Maandblad voor Accountancy
en Bedrijfseconomie, januari/februari 1989. Koninklijke Nederlandsche Machinefabriek v/h
E. H. Begemann, Helmond
Maatschappij van Berkel’s Patent N.V., Rijswijk
Blydenstein-Willink N.V., Enschede
De Boer Winkelbedrijven N.V., Beilen
N.V. Koninklijke Distilleerderijen Erven Lucas Bols,
Nieuw-Vennep
Koninklijke Borsumij Wehry N.V., Den Haag
Koninklijke Boskalis Westminster N.V., Papendrecht
Bührmann-Tetterode N.V., Amsterdam
Center Paros N.V., Rotterdam
N.V. Centrale Suiker Maatschappij, Diemen3
Cindu-Key & Kramer N.V., Uithoorn
DAF N.V., Eindhoven 297 juli/augustus 1990 8 Lid 2 van dit artikel luidt als volgt: ’Lid 1 is niet reeds van
toepassing wanneer de in de consolidatie op te nemen
ondernemingen deels industriële, deels commerciële en deels
dienstverlenende ondernemingen zijn, of wanneer deze
ondernemingen industriële of commerciële werkzaamheden
uitoefenen die betrekking hebben op uiteenlopende produkten
dan wel uiteenlopende diensten verrichten.’ 12 Het zij vermeld dat de resultaten van beide onderzoeken niet
geheel met elkaar overeenstemmen. Zowel het NIvRA als Van
der Tas hebben de jaarrekeningen over 1984 onderzocht. Het
NIvRA echter telde slechts twee gevallen, terwijl Van der Tas vijf
gevallen telde waarin een beroep werd gedaan op de
uitsluitingsclausule wegens afwijkende bedrijfsactiviteiten. Bij Van
der Tas betrof het de volgende ondernemingen: Boskalis, Fokker,
Heineken, Hoogovens en Internatio Muller. De afwijking tussen
de resultaten van de beide onderzoeken werd onder meer
veroorzaakt door verschillen in de onderzoekspopulatie. Zo
maakte de jaarrekening van Boskalis geen deel uit van het
NIvRA-onderzoek. Verder kunnen ook interpretatieverschillen een
rol hebben gespeeld; door Van der Tas werd Heineken genoemd,
dat in het jaarrapport over 1984 het gebruik van de
uitzonderingsclausule wel vermeldde, terwijl die in dat jaar niet
van toepassing was. 18 Zie onder meer NRC/Handelsblad d.d. 12 oktober 1989. Heineken, Hoogovens en Internatio Muller. De afwijking tussen
de resultaten van de beide onderzoeken werd onder meer
veroorzaakt door verschillen in de onderzoekspopulatie. Zo
maakte de jaarrekening van Boskalis geen deel uit van het
NIvRA-onderzoek. Verder kunnen ook interpretatieverschillen een
rol hebben gespeeld; door Van der Tas werd Heineken genoemd,
dat in het jaarrapport over 1984 het gebruik van de
uitzonderingsclausule wel vermeldde, terwijl die in dat jaar niet
van toepassing was. 13 Het betreft hier uitsluitend niet-financiële instellingen. Met
uitzondering van Unigro en Vendex International waren de
aandelen van deze ondernemingen op 31 december 1989 ter
beurze van Amsterdam genoteerd. 14 De naam van deze vennootschap is in 1989 gewijzigd in DAF
N.V.; sedert juni 1989 zijn de DAF-aandelen beursgenoteerd. 15 De solvabiliteitsratio wordt in deze paragraaf gedefinieerd als
het garantievermogen gedeeld door het totaal vermogen. Het
garantievermogen omvat het eigen vermogen en voor zover van
toepassing, het belang derden, de egalisatierekening
investeringspremies en de achtergestelde leningen. 16 Voor de goede orde zij vermeld dat onlangs de helft van de
aandelen Aircraft Financing & Trading zijn verkocht aan Rolls
Royce & Partners Finance (zie onder meer NRC/Handelsblad d.d. 3 februari 1990 en 10 mei 1990). 17 Interessant in dit kader is de kritiek van de
enquêtecommissie OGEM omtrent het niet meeconsolideren van
de meerderheidsdeelnemingen in onroerend goed
maatschappijen door OGEM Holding N.V. De commissie, die in
opdacht van de Ondernemingskamer het beleid en de gang van
zaken bij OGEM in de periode 1970-1983 onderzocht, vond het
bedenkelijk dat de balansverhoudingen bewust gunstiger werden
voorgesteld, onder andere door het (tot 1980) buiten de balans
houden van het zogenaamde bedrijfsgebonden onroerend goed. Ter verbetering van de kwaliteit van de jaarrekening werd door
OGEM besloten om met ingang van 1980 de onroerend goed
maatschappijen mee te consolideren (zie het verslag van de
enquêtecommissie, pagina 105). In het OGEM-jaarverslag over
1980 wordt hierover het volgende gemeld: '...Tot deze wijziging
van de consolidatiegrondslagen is besloten naar aanleiding van
de bij de Tweede Kamer der Staten-Generaal ingediende
Aanpassingswetgeving inzake het vennootschapsrecht
(aanpassing van de wetgeving aan de Vierde EG-richtlijn, RGAV). In deze Aanpassingswetgeving wordt een duidelijke voorkeur
uitgesproken voor consolidatie in de jaarrekening van de
moedermaatschappij van deelnemingen zoals deze onroerend
goed maatschappijen.’ definiëring van de begrippen dochtermaatschappij en
groepsmaatschappij onder de oude en nieuwe wet. Zie hiervoor
onder meer Timmers (1989). 8 Lid 2 van dit artikel luidt als volgt: ’Lid 1 is niet reeds van
toepassing wanneer de in de consolidatie op te nemen
ondernemingen deels industriële, deels commerciële en deels
dienstverlenende ondernemingen zijn, of wanneer deze
ondernemingen industriële of commerciële werkzaamheden
uitoefenen die betrekking hebben op uiteenlopende produkten
dan wel uiteenlopende diensten verrichten.’
Hieruit kan worden afgeleid dat zogenaamde ’gemengde
groepen’, dat wil zeggen groepen met diverse activiteiten, geen
beroep op de uitzonderingsclausule kunnen doen. 9 Memorie van Antwoord d.d. 27 oktober 1987; Tweede Kamer,
vergaderjaar 1987-1988,19813, nr. 5. 10 Raad voor de Jaarverslaggeving, Extra publikatie inzake
Bijzondere bepalingen voor de geconsolideerde jaarrekening en
Enige vereenvoudigingen en verduidelijkingen in het
jaarrekeningenrecht, augustus 1989, pagina 10. 11 Vergelijk artikel 14 lid 2 van de Zevende EG-richtlijn (zie noot
8). definiëring van de begrippen dochtermaatschappij en
groepsmaatschappij onder de oude en nieuwe wet. Zie hiervoor
onder meer Timmers (1989). Literatuur - groepen die de omvang van kleine onderneming niet
overschrijden. Beckman H. (red.), Consolidatiewet en jaarrekening, NIvRA-
geschrift 51, Kluwer, Deventer, 1989. Beckman H. (red.), Consolidatiewet en jaarrekening, NIvRA-
geschrift 51, Kluwer, Deventer, 1989. 4 In Nederland dienen alle deelnemingen waarin men invloed
van betekenis uitoefent op het zakelijke en financiële beleid in
principe te worden gewaardeerd tegen de nettovermogens-
waarde (zie artikel 389 BW 2). Beekman H. & Van Hoepen M. A., Deelnemingen en
groepsjaarrekening, Euroforum, Eindhoven, 1984. Beekman H. & Van Hoepen M. A., Deelnemingen en Beekman H. & Van Hoepen M. A., Deelnemingen en groepsjaarrekening, Euroforum, Eindhoven, 1984. Beekman H. & Volgenant jr. O. M. (red.), Documentatie rond de
jaarrekening, Kluwer, Deventer, bijgewerkt tot en met mei
1989. 5 Door het IASC wordt het begrip ’subsidiary’ als volgt
gedefinieerd: 'A subsidiary is an enterprise that is controlled by
another enterprise (known as the parent).’ Zie paragraaf 3, IAS
22. Enquêtecommissie OGEM (Van der Hoeven J., Van Putten A. &
Slagter W. J.), Verslag van het onderzoek aan de
Ondernemingskamer, 31 december 1986. 6 Brief van de Raad voor de Jaarverslaggeving aan het IASC
d.d. 24 februari 1988. 7 Uit deze formulering kon worden afgeleid dat in principe
dochtermaatschappijen steeds moesten worden Financial Accounting Standards Board, Statement o f Financial
Accounting Standards No. 94: Consolidation of All Majority-
owned Subsidiaries, Stamford, Connecticut, 1987. 7 Uit deze formulering kon worden afgeleid dat in principe
dochtermaatschappijen steeds moesten worden
meegeconsolideerd, dan wel dat slechts groepsmaatschappijen
mochten worden meegeconsolideerd. owned Subsidiaries, Stamford, Connecticut, 1987. meegeconsolideerd, dan wel dat slechts groepsmaatschappijen
mochten worden meegeconsolideerd. Heian J. B. & Thies J. B., Consolidation of Finance Subsidiaries:
$230 Billion in Off-Balance-Sheet Financing Comes Home to
Roost, Accounting Horizons, maart 1989. Op de vraag ten aanzien van welke maatschappijen een
consolidatieplicht dan wel een consolidatiebevoegdheid gold,
wordt niet nader ingegaan. Verwezen zij onder meer naar
Beekman (1989, hoofdstuk I) en Beekman & Van Hoepen (1984,
paragraaf 9.2). Ook zal niet worden ingegaan op de verschillen in Livnat J. & Sondhi A. C., Finance Subsidiaries: Their Formation
and Consolidation, Journal of Business Finance &
Accounting, lente 1986. 298 juli/augustus 1990 18 Zie onder meer NRC/Handelsblad d.d. 12 oktober 1989. Op grond daarvan zou Aircraft
Financing & Trading voortaan niet meer behoeven te worden
meegeconsolideerd. Een en ander doet evenwel geen afbreuk
aan de onderhavige problematiek, namelijk de wijze waarop de
uitsluitingsclausule wegens afwijkende bedrijfsactiviteiten zou
moeten worden geïnterpreteerd. 299 juli/augustus 1990
|
W2316923084.txt
|
https://zenodo.org/records/1947549/files/article.pdf
|
de
|
XXXIII. Ammoniumselenat und die Frage der Isodimorphie in der Alkalisalzreihe.
|
Zeitschrift für Kristallographie. Crystalline materials
| 1,907
|
public-domain
| 9,982
|
XXXIII. Ammoniumselenat und die Frage der
Isodimorphie in der Alkalisalzreihe.
Von
Δ. Ε. H. Tutton in London.
(Mit 9 Textfiguren.)
Ammoniumselenat (NH^SeOt wurde nicht mit den anderen Alkalisulfaten und -selenaten in der letzten Mitteilung über diese Salzreihe (diese
Zeitschr. 1905, 41, 381) behandelt, weil der Verf. es nicht möglich gefunden
hatte, isomorphe rhombische Krystalle dieses Salzes darzustellen. Im Jahre
1862 veröffentlichte Victor von L a n g (Sitzungsber. d. k. Akad. Wiss. Wien
45, Abt. 2, 1 09) eine Beschreibung rhombischer Krystalle von selensaurem
Ammonium, welche von v. H a u e r dargestellt worden waren, und welche
sich isomorph mit denjenigen des schwefelsauren Ammoniums erwiesen.
Ihre Elemente, nach der modernen Axenfolge angeordnet, sind die folgenden:
Beobachtete Formen: {010}, (100), {011}, {103}, {130}, {133}.
Axenverhältnis: a : b : e =
Hauptwinkel :
0,5335 : 1 : 0,7499.
(011 ) : (010) =
(011) : (0T1)
(103): (100)
(103): (103)
(130): (100)
(103): (011)
(133) : (T33)
(133): (103)
53» 29'
73 16
65 27
50 26
58 0
43 34
82 24
34 11
Fig. 1.
Erste Mittellinie parallel {130}, zweite Mittellinie
parallel {011}.
Optisches Zeichen negativ, Dispersion beträchtlich. Scheinbarer optischer Axenwinkel für rot 78° 35', für blau 87° 40'.
von L a n g erwähnte auch einige dünne nadeiförmige Krystalle, welche
G r o t h , Zeitschrift f. Kryetallogr. XLÍI.
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
530
Α. Ε. Η. Tutton.
einen Prismenwinkel von 42° 55' besaßen und optisch verschieden von den
rhombischen Krystallen waren.
Fig. 1 ist eine Reproduction der Zeichnung der rhombischen Krystalle,
mit modernen Flächensymbolen versehen.
Zehn Jahre später stellte T o p s a e das Salz dar (Arch, des Sciences
phys. et nat., Genève 1872, 45, 80J, indem er Selensäure mit Ammoniak
sättigte und die Lösung dem freiwilligen Verdunsten aussetzte. Er erhielt
aber prismatische und tafelförmige monosymmetrische Krystalle und gar
keine rhombischen. Er meinte daher, daß Ammoniumselenat dimorph sei,
daß aber die Bedingungen für die Bildung der rhombischen Modification
unbekannt seien.
Die Möglichkeit der Isodimorphie bei den verschiedenen Salzreihen
von Kalium, Rubidium, Cäsium und Ammonium ist in der letzten Zeit von
G o ß n e r vermutet worden (diese Zeitschr. 1905, éO, 69). Es geschieht bekanntlich häufig, daß die entsprechenden Salze derselben Säure verschieden
unter verschiedenen Bedingungen krystallisieren. In der Tat hatte G o ß n e r
schon angedeutet (ebenda 1904, 39, 381), daß die sauren Sulfate von Kalium
und Ammonium, KIISOi und (NHi)HSOi, in ganz verschiedenen Modiflcationen des rhombischen Systems krystallisieren. Die gleiche Verschiedenheit kommt zwischen den Kalium- und Rubidiumsalzen vor im Falle der
Doppeluranylnitrate, welche von S a c h s untersucht worden sind (ebenda
1904, 38, 496), indem das Kaliumsalz rhombisch und das Rubidium- und
das Gäsiumsalz rhomboëdrisch krystallisieren. Entgegengesetzt kommt sie
zwischen den Rubidium- und Cäsiumsalzen vor bei den Doppelchloriden mit
Eisenchlorid, welche von G o ß n e r in der Mitteilung von 1905 beschrieben
worden sind. In jedem Falle aber, als man die verschieden krystallisierenden Glieder der Reihe aus gemischten Lösungen krystallisierte, erhielt man
die fehlende Krystullform immer in den Mischkrystallen. G o ß n e r kommt
daher zur folgenden Schlußfolgerung: »Die entsprechenden Salze von JVi?4)
Κ, Rb, Os bilden immer eine isomorphe Gruppe, zeigen jedoch auffallend
häufig die Erscheinung der Isodimorphie. In reinem Zustande ist meist
nur die eine Modification für ein Salz bekannt; in isomorphen Mischungen
ist jedoch die fehlende zweite Modification immer nachzuweisen.«
Darstellung des reinen Salzes und seine Krystallisation
unter verschiedenen Bedingungen.
Das Salz ist durch Sättigung reiner Selensäure (von Merck bezogen)
mit Ammoniakgas dargestellt worden ; die Selensäure wurde hierzu mit dem
gleichen Volum Wasser verdünnt, in die drei Kölbchen eines U-Absorptionsrohres gebracht. Die Reaction geht mit beträchtlicher Wärmeentwickelung vor sich. Das Salz ist beinahe zweimal so löslich als Ammoniumsulfat und ist in der Tat auffallend zerfließlich, unter Bildung einer
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
Amnioniumselenat und die Frage der Isodimorphie in der Alkalisalzreihe.
531
sehr viscosen gesättigten Lösung. Vier Präparate sind dargestellt worden,
in jedem Falle ca. 25 g. Die filtrierten Lösungen sind zur Sättigung
oder Übersättigung concentriert und unter den folgenden verschiedenen
Bedingungen auf den Krystallisationspunkt gebracht worden, entweder im
ganzen oder in geteilten Mengen: 1) in den freien Luft des Laboratoriums
(freiwilliges Verdunsten); 2) im Exsiccator über Schwefelsäure bei gewöhnlichem Drucke; 3) über Schwefelsäure im Vacuum; 4) bei warmer Sommertemperatur unter den drei vorgenannten Bedingungen; 5) in ähnlicher
Weise bei kalter Wintertemperatur ; 6) bei der Temperatur einer Kältemischung von Eis und Salz; 7) bei der Temperatur von 70° nach der
Sättigung bei 100°; 8) bei gewöhnlicher Temperatur in Anwesenheit eines
Krystalles von schwefelsaurem Ammonium, welcher in die Lösung gebracht
wurde.
In jedem Falle sind nur monosymmetrische Krystalle gebildet worden,
deren Charaktere später in dieser Mitteilung beschrieben werden und
welche der kurzen Beschreibung von Topswe entsprechen. Bestimmungen
des Selens, durch Reduction mit Salzsäure und Schwefeldioxyd, zeigten, daß
sie aus reinem, wasserfreiem Ammoniumselenat bestanden. Im Falle 8)
löste sich der Krystall des schwefelsauren Ammoniums auf.
Die Wirkung verschiedener Lösungsmittel ist ebenfalls untersucht worden. Alkohol und Aceton lösen das Salz nicht bemerkbar auf, aber wasserfreie Essigsäure ist ein vortreffliches Lösungsmittel. Aus dieser Lösung
schieden sich nur monosymmetrische Krystalle ab, ähnlich, was den Habitus
und die Winkel betrifft, denjenigen welche aus Wasser krystallisieren.
Also gingen alle Versuche der Darstellung einer rhombischen Modification des reinen selensauren Ammoniums vollkommen fehl.
Mischungsversuche.
a) S e l e n s a u r e s u n d s c h w e f e l s a u r e s A m m o n i u m .
Es wurden
zwei Lösungen bereitet, welche gleiche molekulare Mengen der beiden Salze
in dem einen Falle resp. zwei Moleküle des Selenats mit einem Äquivalent
des Sulfats in dem anderen Falle enthielten. In beiden Fällen schieden
sich beim Abkühlen oder Verdunsten rhombische Krystalle ab, welche denjenigen des Sulfats ähnlich waren, und im Falle der gleiche Äquivalente
enthaltenden Lösung war dieses der einzige Typus. Im anderen Falle aber
sind auch monosymmetrische Krystalle gefunden worden, welche ähnlich
denjenigen des selensauren Ammoniums waren. Die Zusammensetzung der
Krystalle von mehreren Krystallisationen ist mit Hilfe von Dichtigkeitsbestimmungen nach der Schwebemethode festgestellt worden, indem man als
Flüssigkeit eine Mischung von Methylenjodid und Benzol brauchte.
Das spec. Gewicht von Ammoniumsulfatkrystallen bei 20°/4° ist schon
früher = 1,772 gefunden worden (diese Zeitschr. 1905, 4 1 , 385), und
34*
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
532
Α. Ε. Η. Tutton.
dasjenige des reinen Ammoniumselenats (monosymmetrisch) ist, wie später
in dieser Mitteilung gezeigt werden wird, = 2,194.
Rhombische Krystalle aus der Lösung von gleichen molekularen Äquivalenten. Spec. Gewicht bei 20°/4°: Krystallisation 1 : 1,813; Krystallisation 2 : 1,822. Diese W e r t e entsprechen Krystallen, welche 9,7 und resp.
11,8 °/ 0 Ammoniumselenat enthalten. Diese Krystalle waren sehr ähnlich
denjenigen von Ammoniumsulfat, und bei der goniometrischen Untersuchung
lieferten sie Winkel, welche nur innerhalb 3' von denjenigen für jenes Salz
verschieden waren.
Krystalle aus der Lösung von (NH t )^SO á + 2 ( N H ^ S e O ^ . Die zwei
Arten von Krystallen hatten die folgenden Dichtigkeiten:
1. Rhombische tafelförmige Krystalle, ähnlich denjenigen aus der Lösung
gleicher Äquivalente: 1,840 bei 20°/4°.
2. Monosymmetrische Krystalle, verlängerte Prismen gleich denjenigen,
welche später in Fig. 4 repräsentiert w e r d e n : 2,066 bei 20°/4°.
Diese Dichtigkeiten entsprechen Krystallen, welche 16,1 und resp.
69,7 °/ 0 Ammoniumselenat enthalten. Vier Krystallisationen wurden untersucht und immer mit demselben Resultate, Mischungen von rhombischen
Platten u n d von Gruppen monosymmetrischer Prismen.
Zwillinge des
letzten Typus sind auch beobachtet w o r d e n , genau ähnlich denjenigen,
welche später f ü r reines Ammoniumselenat zu beschreiben sind.
b) A m m o n i u m - u n d K a l i u m s e l e n a t .
Eine Lösung gleicher molekularer Mengen gab nur rhombische Mischkrystalle des Typus Kaliumselenat
und von ähnlichen Winkeln.
Spec. Gewicht bei 20°/4°: 2,793.
Da das
spec. Gewicht des Kaliumselenats 3,067 ist, enthielten die Krystalle 31,4 °/ 0
Ammoniumselenat.
c) A m m o n i u m - u n d R u b i d i u m s e l e n a t .
Eine Lösung gleicher 1
molekularer Mengen dieser Salze lieferten auch nur rhombische Mischkrystalle von ähnlichem Typus und Winkeln, wie diejenigen des Rubidiumselenats. Spec. Gewicht bei 20°/4° 3,160, 48,4 °/ 0 Ammoniumselenatgehalt
entsprechend (spec. Gewicht des reinen Rb^SeOi 3,902). Eine zweite Lösung
im Verhältnisse 2 Mol. des Ammoniumselenats und ein Molekül des Rubidiumselenats gab ebenfalls nur rhombische Mischkrystalle. Zwei Krystallisationen wurden untersucht; die erste h a t das spec. Gewicht 2,891, 59,19 °/ 0
Ammoniumselenat entsprechend, und die zweite besaß das spec. Gewicht
2,858, 6 1 , 1 2 % entsprechend.
Diese letzteren Mischkrystalle waren besonders vollkommen ausgebildete
rechtwinklige Tafeln parallel è {010}, modiflciert von den Flächen der Prismen ρ {110} und y {130} und den Domen g {011} und q'{021}; sie enthielten m e h r Ammoniumselenat, als irgend andere erhaltene rhombische
Krystalle.
Die Leichtigkeit, mit welcher diese zwei Salze zusammenkrystallisieren,
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
A m m o n i u m s e l e n a t und die Frage der Isodimorpbie in der Alkalisalzreihe.
533
ist zweifellos durch die große Ähnlichkeit veranlaßt, welche, wie der Verf.
gezeigt h a t , im allgemeinen zwischen der Structur der Ammonium- und
Rubidiumsalze existiert, und welche durch ihre beinahe identischen topischen
Axen am klarsten ausgedrückt wird.
Untersuchung der von Langschen Krystalle.
Als Verf. das Glück hatte, mit der Freundschaft des Herrn Prof. v o n
L a n g beehrt zu sein, teilte er ihm die Hauptresultate dieser Untersuchung
mit. Die Krystalle von 1862 waren glücklicherweise vorsichtigst aufbewahrt worden und Herr Prof. v o n L a n g schickte sie freundlichst von Wien
aus dem Verf. nach London.
Sie waren in einem kleinen Glasrohre enthalten, in Baumwolle gepackt,
und das Rohr war durch einen dicht passenden Korkstöpsel geschlossen.
Ihr Aussehen nach 44jährigem Aufbewahren war noch hinreichend gut, um
sehr verwertbare Beobachtungen zu erlauben, obgleich ihre Menge nicht
groß genug für eine Analyse w a r , ohne die gesamte Probe zu zerstören.
Die Krystalle hatten viele ihrer Kanten durch Zerfließen verloren, dessenungeachtet lieferten manche der Flächen ziemlich gute Reflexbilder.
Die meisten der Krystalle entsprachen genau dem rhombischen Typus,
welcher von Herrn Prof. v. L a n g beschrieben worden ist, außerdem aber
befanden sich darunter zwei oder drei der dünnen nadeiförmigen Krystalle,
welche er auch erwähnt. Die am besten erhaltenen der letzteren und
mehrere des rhombischen Typus wurden für die Untersuchung ausgewählt.
Ihre Dichte wurde zuerst mit Hilfe der Schwebemethode bestimmt, und
nachher wurden sie goniometrisch und optisch untersucht. Die Resultate
der Dichtigkeitsbestimmungen sind unten gegeben:
Spec. Gewicht 200/4«:
Rhombische Modification
Verlängertes Prisma (Nadel)
2,0774
2,1719
Also entspricht die Nadel sehr nahe dem monosymmetrischen Ammoniumselenat. Spätere goniometrische Untersuchung zeigte sogleich, daß sie
in der Tat identisch mit der monosymmetrischen Modification T o p s e es
und des Verfs. war. Die verlängerte Prismenzone war [αcm], welche
immer in den Präparaten des Verfs. die vorherrschende war, nach welcher
die Krystalle langprismaliseh waren. Eine zuverlässige Messung von ac =
(100): (001) gab den Winkel 64« 36', eine andere von cm = (001) : (201)
gab 68° 0', und eine dritte von am = (100) : (20T) lieferte 47° 20'. Diese
sind hinreichend nahe den Mittelwerten des Verfs. (s. Tabelle auf S. 536)
für diese Winkel, 64° 31', 68° 22' und 47° 7', um zu beweisen, daß die
nadeiförmigen Krystalle identisch mit der monosymmetrischen Modification
sind. Die Endflächen waren auch hinreichend gut, um die Identificierung
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
534
Α. Ε. Η. Tutton.
mit denjenigen des monosymmetrischen primären Prismas ρ {110} und der
Hemipyramide o'{Tl1} zu ermöglichen. Der Prismenwinkel 42°55', welchen
Herr Prof. v. Lang für die von ihm gemessenen dünnen Nadeln angibt,
war zweifellos derjenige eines Zwillingsprismas, welches Verf. als eine sehr
häufig vorkommende Form gefunden hat, und welches einen theoretischen
Winkel von 43« 16' hat.
Der beste Krystall der rhombischen Modification war eine rechtwinklige rhombische Platte, und, obgleich das Zerfließen etwas begonnen hatte,
ergaben die Messungen derselben doch klar, daß die vorherrschende (tafelförmige) Ebene des Krystalles das Brachypinakoid 6(010} war, daß die
längeren Kanten durch Flächen des Prismas {130} gebildet wurden, ca. 31°
dazu geneigt, und daß die kürzeren Kanten (Zone senkrecht zur eben genannten) durch das Doma {011} gebildet wurden, 53^° zur Tafelebene
geneigt. Diese Werte sind ganz hinreichend, um die Identificierung des
Krystalles mit einem derjenigen zu gestatten, welche von Herrn Prof.
v. L a n g gemessen worden sind, und sie bestätigen die rhombische Symmetrie und die Ähnlichkeit mit dem Ammoniumsulfat und den anderen
Alkalisulfaten und -selenaten. Die Stellung und der Winkel der optischen
Axen wurden durch Eintauchen des Krystalles in Anisöl bestimmt, welches
Öl beinahe denselben Brechungsexponenten hat.
Die erste Mittellinie ist die Normale zu c{001} und die Ebene der
optischen Axen ist die Tafelebene δ {010}. Der wahre Axenwinkel 2 Va
für rotes Licht ist 55° 36' und für Blau 58° 54'. Also besitzen Ammoniumsulfat und -selenat eine identische Axenebene, jedoch, wie Herr Prof. v. Lang
angibt, sind die erste und zweite Mittellinie vertauscht.
Was die Zusammensetzung der v. Lang sehen Krystalle betrifft, so
deuten die Dichtigkeiten an, daß die Mutterlauge etwas Ammoniumsulfat
enthielt; denn die Resultate entsprechen rhombischen Krystallen, welche
7 3 % Ammoniumselenat und 2 7 % Ammoniumsulfat enthalten, und monosymmetrischen Krystallen, welche 95 % Ammoniumselenat enthalten. Diese
Zahlen für die Verunreinigung sind aber wahrscheinlich ein wenig größer,
als es der Wahrheit entspricht, weil innere Zerfließlichkeit, aller Wahrscheinlichkeit nach, etwas mitzurechnen ist, und diese würde vielleicht die
fehlenden 5 % bei den monosymmetrischen Krystallen, als auch vielleicht
eine entsprechende Menge bei den rhombischen Krystallen erklären. Es
ist aber gewiß, daß die Resultate a) die Anwesenheit einer isomorphen
Beimischung beweisen, und zwar am wahrscheinlichsten aus Ammoniumsulfat bestehend, weil dieses das einzige Salz der Reihe ist, welches ein
niedrigeres speciflsches Gewicht besitzt, und b) daß der größere Teil (beinahe alles) dieser Verunreinigung in den rhombischen Krystallen concentriert worden ist, welche streng isomorph mit dem Ammoniumsulfat sind.
Es ist schon (S. 532) gezeigt worden, daß rhombische Ammoniumselenat
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
A m m o n i u m s e l e n a t u n d die Frage der Isodimorphie in der Alkalisalzreihe.
535
enthaltende Mischkrystalle leicht gewonnen werden, wenn 33 °/0 eines rhombischen Salzes der Reihe neben Ammoniumselenat in der Mutterlauge anwesend sind, und solche Mischkrystalle sind in der Tat vom Verf. mit einem
Gehalte von etwas mehr als 60 °/0 an Ammoniumselenat erhalten worden.
Die von H a u e r sehen rhombischen Krystalle sind noch etwas reicher (75 °/0)
an Ammoniumselenat, die Differenz ist aber relativ so gering, daß gar kein
Zweifel sein kann, daß ein 75°/ 0 enthaltender Mischkrystall ganz möglich
ist, und zwar daß hier die Erklärung der Bildung der von H au ersehen
rhombischen Krystalle liegt.
Monosymmetrische Modification dee Ammoniumselenats.
Eine Bestimmung des Selens in 1,0406 g der Krystalle, durch Reduction
mit Salzsäure und Schwefeldioxyd, gab 43,72°/ 0 Se. Berechnet 44,16.
Es wurde etwas schwer gefunden, die letzten Spuren des Selens niederzuschlagen. T o p s 0 e erhielt auch nur 43,7.
S y m m e t r i e : Monosymmetrisch holoedrisch.
H a b i t u s : Prismatisch (häufig acicular) längs der Richtung der Symmetrieaxe b, oder tafelförmig parallel der Basis, dem Orthopinakoid oder
der Hemipyramide o'{Tl1).
S t r u c t u r t y p u s : Pseudohexagonal, primäre Prismenzonenflächen ca.
60° (innerhalb 1°) geneigt.
A x e n v e r h ä l t n i s : a: b : c = 1,8900 : 1 : 1,1987.
A x e n w i n k e l : β = 64» 31'.
B e o b a c h t e t e F o r m e n : «{100}, c{001), m{201}, p{110}, / { 3 1 0 } ,
o{111}, ο'{111}, M{5H}.
Z w i l l i n g e : Sehr häufig beobFig. 2.
achtet; Zwillingsebene senkrecht zur
Symmetrieebene und zur Normale
zu c{001}, welche letztere die Zwillingsaxe ist.
S p a l t u n g s r i c h t u n g e n : Drei
Richtungen, parallel a {100} am
aifooi
vollkommensten, c{001} am nächsten in der Ordnung der Vollkommenheit, und δ {010}.
Die stereographische Projection
ist in Fig. 2 gegeben.
Zweiundzwanzig Krystalle sind
bei der goniometrischen Arbeit angewandt worden, aus elf verschiedenen Krystallisationen ausgewählt, von
welchen die einen aus gesättigten Lösungen im Vacuum über Schwefelsäure
sich gebildet hatten, gewisse andere über Schwefelsäure bei dem gewöhn-
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
Α. Ε. Η. Tutton.
536
lichen Drucke krystallisiert waren und zwei übrigbleibende aus stark übersättigten Lösungen an der freien Luft sich abgeschieden hatten.
Die letztere
Methode liefert gute Krystalle
wegen der
nur an sehr trockenen Tagen,
schnellen Zerfließlichkeit der Krystalle in feuchter Luft.
Der prismatische Typus variierte von
Fig. 3.
dünnen
Nadeln
bis zu
dicken Blöcken,
wie in Fig. 3 gezeigt wird ; sie waren aber
sehr häufig von dem in Fig. 4 repräsentiertem Charakter, bei welchem die zwei
Flächen
jeder
Form
der
verlängerten
Zone in sehr verschiedenem Betrage entwickelt waren.
Gewisse der im Vacuum
erhaltenen Krystallisationen lieferten dicke
tafelförmige Krystalle der in Fig. 5 illustrierten Art.
Die relative Entwickelung der Flächen
war so verschieden,
daß jede der fünf Hauptformen « { 1 00}, e { 0 0 1 ) , m { 2 0 1 ) , p { 1 1 0 } ,
o'{T11}
nach einander vorherrschten, und häufig waren die zwei Hälften desselben
Krystalles verschieden
entwickelt.
Fig. 4.
Fig. 5.
Die Krystalle haben ein helles, klares, wachsartiges Ansehen und sind
sehr weich.
Ihre gesättigte wässerige Lösung ist sehr viscos.
Die Resultate der Messungen sind in der folgenden Tabelle zusammengestellt.
Morphologische Winkel
Zahl der
Messungen :
des s e l e n s a u r e n
Ammoniums.
Mittel beob. : Berechn. : Did.
Grenzen :
ac
= (100) :: (001)
40
64° 2 2 ' — 6 4 ° 44'
64031'
cm
= (001): : (201)
39
68 11 — 6 8
42
68 27
68°22'
5'
ma
= (201) :: (T00)
31
46 40 — 4 7
29
47
47
7
5
2
2
—
—
ap'
(100) ::(310)
4
' 29 27 — 2 9
49
29 36
29 38
p'p
ap
- (310): : (110)
= (100) :: (110)
3
29 5 2 — 3 0
8
30
0
30
36
59 23 — 5 9
56
59 38
—
pp
= (110) :: (T10)
13
60 33 — 6 0
50
60 42
0
60 44
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
0
—
2
A m m o n i u m s e l e n a t und die F r a g e der I s o d i m o r p h i e in der Alkalisalzreihe.
Zahl der
Grenzen:
Messungen:
ao =
oo' =
ao' =
o'n =
na =
o'a =
CO =
op
cp po' =
o'c =
mo'
o'p
pn
nm pm =
537
Mittel b e o b . : B e r e c h n . : Dill. :
(100)
(111)
17
52° 4 5 '— 5 3 «
5'
52°58'
52056'
2'
(111)
(111)
14
45
6 — 4 5 29
45 16
4 5 18
2
(100)
29
98
4 — 9 8 25
98 14
9 8 14
0
(TU)
(T11)
(311)
—
38
2
—
(311)
(100)
—
43 44
—
(T00)
26
(TM)
—
—
81 35 — 8 1
58
81 46
81 46
0
(001)
(111)
14
43 43 — 4 3 5 8
43 51
43 53
2
(III)
(110)
14
33 20 — 3 3 43
33 30
33 33
3
(001)
(110)
(11 Τ)
77 11 — 7 7 37
7 — 4 3 34
43
77 2 2
43 19
77 2 6
43 16
4
(110)
34
24
(11Ï)
(001)
31
59 10 — 5 9 27
59 18
(201)
(TU)
29
55 42 — 5 5 56
55 50
55 47
3
54
54 18
54 20
2
34 34
3 5 18
34 35
3 5 18
1
0
69 52
69 53
1
(110)
(311)
(201)
25
(110)
(31T)
(110)
(20T)
23
(TU)
1
1
5 — 5 4 32
—
—
69 30 — 7 0
3
Gesamtzahl der Messungen:
Der einzige von T o p s t f e
448.
gegebene Winkelwert
(der Axenwinkel ß), welchen er 64° 2 7 ' fand.
3
—
ist derjenige für
ae
Die Übereinstimmung liefert
zufriedenstellenden Beweis der Identität der Krystalle.
Die zahlreichen Zwillinge
werden
durch
die Anwesenheit von zwei
verschiedenen Winkeln in der verlängerten Orthoprismenzone [ a e m ]
rakterisiert, anstatt der zwei gleichen
Winkel am
=
47° 7', nämlich aa =
50° 5 8 ' und m w =
4 3 ° 1 6 ' , an
Fie
b'
cha-
fi
'
ver-
schiedenen Seiten von der Mitte. Außerdem zeigen die Enden der verlängerten
Prismen einspringende AVinkel, indem
der Winkel
zwischen
den
zwei
zu-
sammenstoßenden j)-Flächen der beiden Individuen 2 5 ° 8'
zwischen
den
zwei
und
derjenige
o'- Flächen
ver-
schiedener Individuen 61° 2 4 ' beträgt.
Häufig zeigen sich auch Vertiefungen
längs den
flacheren
Kanten der ver-
längerten Prismen, durch das Auftreten
von Streifen der nächsten gewöhnlichen
Flächen jedes Einzelkrystalles veranlaßt.
F ü n f solchc Zwillinge sind ge-
messen worden, und die beobachteten Werte für diese vier Zwillingswinkel
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
538
Α. Ε. Η. Tutton.
waren im Durchschnitte innerhalb 4' mit den berechneten eben jetzt gegebenen Winkeln übereinstimmend. Fig. 6 dient zur Erklärung des Baues
der Zwillinge; die Projectionsebene ist die Symmetrieebene, wie im Falle
der stereographischen Projection der normalen einfachen Krystalle, welche
in Fig. 2 gegeben worden ist.
Es ist als wenn ein Krystallindividuum längs einer Ebene parallel e{001}
durchgeschnitten und nachher eine Hälfte um 180° um die Normale zu
c {001} als Zwillingsaxe umgedreht worden wäre.
In der oben gegebenen Beschreibung sind die verticale und die geneigte Axe c und a der Tops0eschen Beschreibung vertauscht worden.
Dieses ist geschehen, um eine auffallende Ähnlichkeit zwischen der primären
Prismenzone dieser monosymmetrischen Krystalle des selensauren Ammoniums
und der primären Prismenzone der rhombischen Krystalle des schwefelsauren Ammoniums und der Sulfate und Selenate des Kaliums, Rubidiums
und Cäsiums deutlich zu zeigen.
In beiden Fällen findet eine große Annäherung an hexagonale Symmetrie statt. Der Winkel ap = (100) : (11 0) des selensauren Ammoniums
ist 59°38', ap' = (100) : (310) ist 29038' und p'p = (310) : (110) ist 30»0';
die entsprechenden Winkel bei Ammoniumsulfat sind ap = (100) : (110) =
290 24' und ^ = (110) : (130) = 300 0', zusammen = ap' = (100) : (130)
= 59°24'. Die Winkel in dieser Zone sind also in beiden Fällen sehr nahe
30° und 60°, jedoch entspricht das primäre Prisma des einen dem tertiären
Prisma des anderen. Folglich, während die Basis bei dieser (monosymmetrischen) Modification des selensauren Ammoniums geneigt ist, bleiben doch
die Winkel zwischen den Flächen der Prismenzone praktisch dieselben wie
bei der rhombischen Modification.
Diese Art der Aufstellung für die Krystalle ist daher angewendet
worden als am wahrscheinlichsten die richtigste, was die Structur betrifft,
und die topischen Axenverhältnisse sind auf der entsprechenden Voraussetzung von pseudohexagonaler Symmetrie berechnet worden, genau ähnlich den Fällen der anderen Sulfate und Selenate der Alkalireihe, wie sie
in der Mitteilung des Verfs. in dieser Zeitschrift von 1905 gegeben worden
sind (41, 386).
Volum und Structur.
S p e c i f i s c h e s G e w i c h t . Die folgenden sechs Bestimmungen wurden
mit Hilfe der Schwebemethode ausgeführt, indem eine Mischung von
Methylenjodid und Benzol als Eintauchungsflüssigkeit benutzt wurde.
Kleine und gut ausgebildete, im Vacuum gewachsene Krystalle wurden
angewandt.
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
Ammoniumselenat und die Frage der Isodimorphie in der Alkalisalzreihe.
Bcobachtungstemperatur:
Entsprechendes
spec. Gewicht mit Wasser
bei 4° verglichen:
Berechnetes
spec. Gewicht
2,1937
2,1934
2,1944
2,1933
2,1940
2,1939
2,1931
18,40
19,4
17,7
14,6
14,5
15,0
für
2,1945
2,1938
2,1952
2,1951
2,1942
Mittel :
539
200/40
2,1937
Die Dichtigkeit des selensauren Ammoniums, krystallisiert in der gewöhnlichen monosymmetrischen Gestalt, ist also 2,194 für 20°/4° angenommen worden.
Die Stellung des selensauren Ammoniums in der Selenatreihe, was die
Dichtigkeit betrifft, ist ähnlich derjenigen des schwefelsauren Ammoniums
in der Sulfatreihe, wie klar aus der folgenden Tabelle zu ersehen ist, denn
jedes ist das leichteste Glied der betreffenden Reihe.
{NH^SOt
ίΓ 2 £0 4
Rb^SOi
Cfc2S04
1,772
2,666
3,615
4,246
M
Molekularvolum — =
(NH^SeOt
K2SeOt
R^SeOi
Gs^SeOt
177 98
^
= 81,12.
2,194
3,067
3,902
4,456
Es wird aus der folgenden
Tabelle der Molekularvolumina der acht Salze der Reihe klar hervorgehen,
daß die Stellung des selensauren Ammoniums analog derjenigen des schwefelsauren Ammoniums ist, nämlich sogleich nach dem Rubidiumsalz.
KjSOi
Rb^SOi
(NHJiSOi
Cs 2 S0 4
64,91
73,34
74,04
84,58
K2Se04
Rb^SeOi
(iV7J4)2Se04
Cs^SeOi
71,67
79,94
81,12
91,09
T o p i s c h e A x e n v e r h ä l t n i s s e . Diese sind sowohl für die Voraussetzung der gewöhnlichen monosymmetrischen Structur berechnet worden,
als auch für diejenige von pseudohexagonaler Structur, und zwar die
letztere aus den Gründen, welche schon erklärt worden sind. Nach der
ersteren Voraussetzung repräsentieren χ, xp und ω die relativen Entfernungen der Centren der Gleichgewichtslagen der molekularen Structureinheiten ( S o h n ck e sehen Punkte) längs der Richtungen der drei monosymmetrischen Axen. Sie sind durch die folgenden Formeln gegeben, wobei
a, b, c die morphologischen Axenverhältnisse sind, β der Winkel zwischen
den geneigten Axen a und e und V das Molekularvolum ist:
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
Α. Ε. Η. T u t t o n .
540
Χ
_
V/
' β sin ß '
a W
Ihre Werte sind:
Fig. 7.
2
ψ
XI)
ψ sin '
'
ac
ß
—
Ϋ/
'
sin '
C W
r
=
ω
a
ß
χ : ψ : ω = 6,4456 : 3,4104 : 4,0880.
Für die zweite (pseudohexagonale) Voraussetzung ist die Structuranordnung wie in
Fig. 7 gezeigt wird, welche einen Durchschnitt
durch den Krystall senkrecht zur primären
Prismenzone [apb] gibt, xp repräsentiert die
Entfernung der Structurpunkte längs der Symmetrieaxe b, und ω diejenige längs der Verticalaxe c ; χ repräsentiert aber die Entfernung
nicht längs der Axe α, sondern längs zweier
pseudohexagonaler Axen, welche zur Ebene
der Figur geneigt sind, und deren zwei in der
Zeichnungsebene liegenden und sie repräsentierenden Spuren ca. 60° zu einander und zu
der Symmetrieaxe b geneigt sind. Sie werden
durch die folgenden Formeln ausgedrückt:
' a c sin/î
r
aesmß'
r
asm
ß'
und ihre numerischen Werte sind:
χ:ψ :ω =
4,5939 : 4,2968 : 5,1506.
Es ist interessant, diese topischen Parameter des monosymmetrischen
selensauren Ammoniums mit denjenigen der rhombischen Sulfate und Selenate der Alkalien zu vergleichen, welche analog in der früheren Mitteilung
des Verfs. (diese Zeitschr. 1905, 4 1 , 386) pseudohexagonal ausgedrückt
worden sind. Die topischen Parameter des Ammoniumselenats sind sehr
ähnlich denjenigen, welche in jener Tabelle gegeben werden. Die Werte
von φ und ω liegen zwischen denjenigen für Kalium- und Cäsiumselenat,
und in der Tat ist ψ nahe an dem Werte des Cäsiumsalzes und ω an dem
des Kaliumsalzes. Das weniger streng vergleichbare Parameterpaar χ ist
von derselben Ordnung, wie die χ-Werte für die metallischen Selenate,
und zwar etwas größer als der Wert für Cäsiumselenat. Obgleich die
einzelnen Parameter natürlich nicht so streng vergleichbar sind, wie diejenigen der rhombischen Salze unter einander, so ist es doch von Wichtigkeit, daß der Durchschnittswert (Mittel von χ , ψ und ω) der topischen
Parameter des selensauren Ammoniums eine mittlere Stellung zwischen den
Durchschnittswerten für die Rubidium- und Cäsiumsalze einnimmt.
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
A m m o n i u m s e l e n a t und die Frage der Isodimorphie in der Alkalisalzreihe.
541
Löslichkeit.
Drei Bestimmungen der Löslichkeit der monosymmetrischen Krystalle
von Ammoniumselenat sind ausgeführt worden für die Sättigung bei 7°,
59° und resp. 100°.
B e s t i m m u n g der Löslichkeit des A m m o n i u m s e l e n a t s .
Menge der
Temperatur
Menge des
gesättigten
der
wasserfreien
w e l c h e v o n 100 Teilen
Lösung :
Sättigung:
Salzes:
Wasser gelöst werden:
14,97
26,01
16,46
70
59
100
8,06
16,17
10,91
1 17
164
197
Gewichtsteile des Salzes,
Also löst Wasser bei der gewöhnlichen Temperatur etwas mehr als
sein eigenes Gewicht des selensauren Ammoniums und bei der Siedetemperatur beinahe genau zweimal sein eigenes Gewicht.
Die Resultate werden graphisch in der längeren Curve AB in Fig. 8
ausgedrückt, welche beinahe eine gerade Linie ist. Die kürzere Curve CD
Fig. 8.
—
—
—
—
—
—
"Ί
—
....
I
ιι
—
- - —
—
—
y
-
—
4
/
•
/
—
/
I""
e
/
/
D y /
y
/
y
—
—
-
-
/
//
y
/
<
Temperatur
10°
ist die Überlöslichkeitscurve, welche freundlichst von Herrn Prof. M i e r s
und Miss I s a a c bestimmt worden ist, mit Hilfe der Refractionsmethode,
welche sie in der letzten Zeit beschrieben haben (Journ. Chem. Soc. 1906,
89, 413). Sie ist aus den Daten der Tab. S. 542 oben construiert worden.
Also läuft die Überlöslichkeitscurve beinahe parallel und bei den niedrigeren Temperaturen in der Nähe der gewöhnlichen sehr nahe der gewöhnlichen Löslichkeitscurve ; die zwei Curven divergieren aber von ungefähr
20° ab mit sich vergrößernder Schnelligkeit. Folglich gibt es nur eine
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
Α. Ε. Η. Tutton.
542
Salzteile zu
Maximalwert des
Temperatur bei
100 Teilen Wasser:
Brechungsexponenten :
d e m Maximum:
142,5
130,22
126,8
124,35
1,42528
1,42192
1,42127
1,42107
25°
19,5
16,2
14
schmalere Zone der Metastabilität zwischen dem ungesättigten und dem
labilen Zustande bei der gewöhnlichen Temperatur, während diese Zone
beträchtlich breiter bei einer warmen Sommertemperatur ist.
Wenn man dieses Resultat mit denjenigen vergleicht, welche für die
metallischen Selenate (diese Zeitschr. 1898, 29, 67) gefunden worden sind,
und für dieselbe Temperatur von 12°, so findet man, daß 100 Gewichtsteile Wasser bei 12° 115 Teile des selensauren Kaliums, 122 Teile des selensauren Ammoniums, 159 Teile des selensauren Rubidiums und resp. 245 Teile
des selensauren Cäsiums auflösen. Die Löslichkeit des Ammoniumselenats
ist also nur ein wenig größer als diejenige von Kaliumselenat ; sie ist aber
beinahe zweimal so groß als diejenige des schwefelsauren Ammoniums
(74 Teile in 100 des Wassers).
Optische E i g e n s c h a f t e n .
Fig. 9.
(fOO)
(100),
Mittel
O r i e n t i e r u n g der Axen
des optischen
Ellipsoids.
Drei Bestimmungen der Auslöschungsrichtungen in der Symmetrieebene wurden mit drei Schnittplatten ausgeführt, welche parallel
jener Ebene geschliffen waren.
Eine der beiden Auslöschungsrichtungen ist sehr nahe normal zu
c{001}, wie in Fig. 9 gezeigt wird,
und die genauen Stellungen, welche
für sie gefunden worden sind, sind
die folgenden:
1 0 49' hinter der Normale zu c {001}
2 9
2 56
2° 18'
Diese Richtung in der Symmetrieebene ist die mittlere Axe β der
Indicatrix oder b des optischen Velocitätsellipsoids, senkrecht zur optischen
Axenebene, welche letztere also senkrecht zur Symmetrieebene ist. Die andere
Auslöschungsrichtung in der Symmetrieebene (senkrecht zur gemessenen)
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
A m m o n i u m s e l e n a t und die Frage der Isodimorphie in der Alkalisalzreihe.
543
ist die erste Mittellinie und Axe γ der Indicatrix oder c des optischen
Velocitätsellipsoids. Sie liegt innerhalb eines halben Grades mitten zwischen
den Normalen zu a {100} und m {20T}, und zwar bildet sie 23° Ή ' mit der
ersten Normale und 23° 56' mit der letzteren. Die Symmetrieaxe b ist die
zweite Mittellinie und Axe a der Indicatrix oder α des Velocitätsellipsoids.
Das Zeichen der Doppelbrechung ist positiv.
B r e c h u n g s e x p o n e n t e n . Diese sind mit sechs 60°-Prismen bestimmt
worden, von welchen jedes so geschliffen worden ist, daß es zwei Indices
direct liefert. Die Resultate werden in der folgenden Tabelle gegeben.
B r e c h u n g s e x p o n e n t e n des Ammoniumselenats.
Index :
Natur des Lichtes:
Grenzen :
Mittel:
1,5561
Li
1,5554—68
1,5571
1,5565—79
G
Schwingungsrichtung
1,5607
1,5598—613
Na
parallel
1,5641
1,5634—48
Tl
der Symmetrieaxe
1,5687
1,5681—90
F
(zweite Mittellinie).
1,5752
G
1,5744—56
1,5584
Li
1,5576—88
fi
1,5594
G
1,5587—99
Schwingungsrichtung
1,5630
Na
1,5623—35
parallel der Richtung
1,5665
Tl
1,5659—70
in der Symmetrie1,5713
F
1,5706—19
ebene 2° 18' hinter der
1,5781
1,5775—83
G
Normale zu c{001).
1,5796
Li
1,5787—805
1,5806
1,5797—815
C
1,5846
Schwingungsrichtung
1,5839—55
Na
1,5885
parallel der
1,5877—92
Tl
1,5935
ersten Mittellinie.
1,5929—42
F
1,6005
1,6000—12
G
Mittelwert von α, β und γ für JVa-Licht 1,5694.
Der /Î-Index, auf das Vacuum corrigiert (Correction = + 0,0004), für
Licht irgend einer Wellenlänge λ bis zu derjenigen der grünen Thalliumlinie,
wird absolut von der folgenden Formel repräsentiert, welche auch approximativ für das blaue Ende des Spectrums gültig ist:
988 153
6 585 000 000 000
β = 1,5404 +
+ ·
V·
λ*
Die α-Indices werden auch sehr nahe reproduciert, wenn man die
Constante 1,5404 um 0,0024 vermindert, und die y-Indices, wenn man sie
um 0,0216 vergrößert.
Die D o p p e l b r e c h u n g
Für Ii-Licht
C- - Na- -
0,0235
0,0235
0,0239
α
ist wie folgt:
Für 77-Licht
- F- - G- -
0,0244
0,0248
0,0253
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
Α. Ε. Η. Tutton.
544
Brechungsexponenten
bei h ö h e r e n T e m p e r a t u r e n .
Diese
wurden mit zwei der Prismen bestimmt, welche a und β resp. β und γ
lieferten. Bestimmungen wurden nicht nur bei 80° gemacht, wie im Falle
des Ammoniumsulfats, sondern auch bei 100°. Denn als man das a und
β gebende Prisma erhitzte, beobachtete m a n , daß die zwei diesen Indices
entsprechenden Spaltbilder, welche schon bei der gewöhnlichen Temperatur
ausnahmsweise nahe zusammen waren (Entfernung n u r 13' f ü r iVa-Licht),
sich einander noch mehr n ä h e r t e n , so daß sie bei 80° nur um 6' getrennt w a r e n , und in der Tat überlagerten sich die breiten Enden der
W e b s k y sehen Spaltbilder, w ä h r e n d die schmalen centrischen Teile einander
beinahe berührten. Nach der Bestimmung bei 80° wurden die P h ä n o m e n e
weiter verfolgt, und bei 87° hatten die Bilder sich auf 3' g e n ä h e r t , bei
91° wurde die Entfernung nur 2', bei 96° ca. 1' und bei 100° waren die
Bilder für alle Wellenlängen scheinbar identisch. Bei dieser Temperatur,
f ü r welche eine Reihe von Bestimmungen ausgeführt worden sind, hatte
man die (für einen zweiaxigen Krystall) außerordentliche Erscheinung eines
scheinbaren einfachen Signalbildes, welches bei allen Stellungen des Ocularnicols unausgelüschl blieb. In der Tat w u r d e ein Bild ausgelöscht, während
das andere an derselben Stelle im Gesichtsfelde erschien. Über 110° begannen die beiden Bilder sich b e m e r k b a r wieder zu t r e n n e n , nach der
anderen Seile hin, so daß « jetzt β ward und vice versa.
Dieser Identitätszustand zweier der Brechungsexponenten, und also die
temporäre Veränderung des dreiaxigen optischen Ellipsoids in ein Umdrehüngsellipsoid (bei welchem zwei der drei IJauptaxen gleich sind), liefert,
wie von dem Verf. bei a n d e r e n , f r ü h e r bei den Alkalisalzen beobachteten
Fällen fand, die Erklärung des Phänomens der Dispersion der optischen
Axen in gekreuzten Axenebenen ; denn solcher Identitätszustand zweier Indices wird immer von dem P h ä n o m e n gekreuzter Axenebenen begleitet in
allen fünf Fällen, welche jetzt in der Alkalireihe beobachtet worden sind.
Es wird später gezeigt werden, daß Ammoniumselenat streng dieser ltegel
folgt, indem es auch das Phänomen zeigt.
Entsprechende Bestimmungen bei 80° und 100° sind auch mit dem
zweiten Prisma gemacht worden, welches β und γ lieferte, und die folgende
Tabelle gibt die vereinigten Resultate beider Bestimmungsreihen:
Brechungsexponenten für 80»
lìrechungsexponenten für 100°
Natur des Lichtes:
<t
A
y
h und ¡1
γ
Li
G
Nu
TI
F
1,5556
1,5ö63
1 ,5599
1.,5637
1,5682
1,5560
1,5568
1,5605
1,5643
1,5689
1,5764
1,5773
1,5812
1,5852
1,5900
1,5553
1,5561
1,5598
1,5636
1.5682
1,5757
1,5765
1,5804
1,5843
1,5890
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
A m m o n i u m s e l e n a t und die Frage der Isodimorphie in der Alkalisalzreihe.
545
O p t i s c h e r A x e n w i n k e l . Drei Paare von Schnittplatten senkrecht
zur ersten und zweiten Mittellinie sind mit Hilfe des Schneid- und des
Schleifgoniometers hergestellt worden, und sie lieferten die folgenden Messungen, bei welchen α-Bromnaphtalin als Eintauchflüssigkeit für die Bestimmungen von 2 H a und 2H 0 diente.
S c h e i n b a r e r W i n k e l in L u f t , 2E.
Licht:
Platte 1:
Platte 2:
Platte 3:
Mittel
iE:
Li
C
Na
Tl
F
580 20'
58 53
60 15
61 2
62 1
59035'
59 43
61 15
61 54
62 36
59° 9'
59 30
60 43
61 43
62 49
590
1'
59
60
61
62
22
44
33
29
W a h r e r o p t i s c h e r A x e n w i n k e l von A m m o n i u m s e l e n a t ,
Nr. der
Platte
senkrecht
zur ersten
Mittellinie:
Licht:
Nr. der
Platte
Beobachtet
senkrecht
2H„:
zur
zur zweiten
zweiten
Mittellinie:
Beobachtet
Ü.H„:
Berechnet
<2V„ :
2Va.
Mittel
2 V„ :
Ί
ι
2
3
35° 8'
34 52
34 46
1a
2a
3a
126030'
126 5
126 27
37»21
37 10 1\ 370 10'
37 0 I
1
2
3
35 13
35 1
34 55
1a
2a
3a
126 20
125 52
126 17
37 28 )
37 20 \ 37 19
37 10
Na
1
2
3
35 44
35 25
35 2b
1a
2a
3a
125 17
124 45
125 27
38 6
37 54
37 47
Tl
1
2
3
36 0
35 42
35 39
1a
2a
3a
124 10
123 30
124 31
38 33 I
38 22 \ 38 22
38 10
F
\
2
3
36 7
35 56
35 48
1a
2a
3a
122 57
122 13
123 16
38 52
38 49
38 30
I
Li
37 54
38 44
H o r i z o n t a l e D i s p e r s i o n d e r e r s t e n M i t t e l l i n i e . Da die erste
Mittellinie in der Symmetrieebene liegt, während die zweite Mittellinie mit
der Symmetrieaxe identisch ist, wird die erste allein dispergiert. Der Betrag ist bestimmt worden, indem man die Schnittplatte vertical anstatt
horizontal stellte, und die Niçois unter 90° und 0° anstatt 45° und 135° anordnete. Jede optische Axe wurde in das Centrum des Fadenkreuzes gebracht mittels der cylindrischen Justierungsbewegung des Polarisationsgoniometers (am F u e s s s c h e n größeren Axenwinkelapparate), und es sind
alsdann Messungen der seitlichen Stellungsdifferenz der Axe für die gewöhnlichen sechs Wellenlängen des Lichtes gemacht worden, indem die
G r o t h , Zeitschrift f. Kiystallogr. XL1I.
35
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
Α. Ε. Η. Tutton.
546
Platte in 1) Cassiaül und 2) Anisül eingetaucht wurde, deren Brechungsindices etwas höher und resp. niedriger als der Mittelindex der Krystalle
sind. Die beiden Bestimmungsreihen stimmten überein, indem sie andeuteten, daß die erste Mittellinie so dispergiert ist, daß sie für blaues Licht
näher an der Normale zu a{i 00} als für Rot liegt. Der Betrag ist ein
solcher, daß jede der zwei optischen Axen um 12' zwischen G- und i - L i c h t
dispergiert ist, was einer Dispersion der ersten Mittellinie von 15' entspricht.
W i r k u n g e i n e r T e m p e r a t u r e r h ö h u n g auf d e n o p t i s c h e n A x e n w i n k e l . Die Brechungsresultate hatten die Möglichkeit einer Dispersion in
gekreuzten Axcnebenen angedeutet für eine Temperatur in der Nähe von
100°. Schniltplatten 1 und 2 sind mit llilfe des Erhitzungsapparates des
größeren Axenwinkelapparates untersucht worden, mit praktisch identischen
Resultaten, welche zeigen, daß die Voraussetzung fest begründet war. Die
optischen Axen (um 59°—62° getrennt bei gewöhnlicher Temperatur) näherten
sich einander, als die Temperatur stieg. Bei 60° wurde 2 i ? auf 45° vermindert für die Mitte des Spectrums, und bei 75° auf 40°. Bei 90° war
der Axenwinkel nur 30°, und bei 100° wurde er bis zu 23° vermindert.
Weitere Erhitzung verursachte, daß die Hyperbeln sich zu einem einaxigen
rechtwinkligen Kreuz vereinigten, und die Lemniscaten und elliptischen
Ringe in Kreise übergingen, für jede Wellenlänge nach einander von Rot
ausgehend, bei successiv höheren Temperaturen. Die letzteren sind in der
folgenden Tabelle gegeben und sind für die Wärmeleitung des Krystallhalters aus Platin corrigirt worden.
C o r r i g i e r t e T e m p e r a t u r e n f ü r d i e B i l d u n g des e i n a x i g e n
Kreuzes.
Für Li-Licht
C- - N a -
110°
11 q
-
114
-
Tl- -
116
-
F-
119
-
Nach der Bildung des Kreuzes für jede Wellenlänge trennten sich die
Axen längs des verticalen Diameters des Gesichtsfeldes, indem statt der
Symmetrieaxe b die dritte, zu c{00l} beinahe normale Auslöschungsrichtung die zweite Mittellinie wurde und also die optische Axenebene in die
Symmetrieebene überging.
Axen des o p t i s c h e n
Ellipsoids.
Axen der Indicatrix
a :β :γ =
0,9985 : 1 : 1,0138
Axen des optischen Velocitätsellipsoids
α : t) : c =
1,0015 : 1 : 0,9804
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
A m m o n i u m s e l e n a t u n d die F r a g e d e r I s o d i m o r p h i e in der Alkalisalzreihe.
547
Molekulare optische Constanten.
Axe der optischen Indicatrix :
Specifische Refraction
ffi — 1
(w2 +
Molekulare Refraction
-
rfi — 1
M _
2
η +2
d
Specifische Dispersion n G — tic
Molekulare Dispersion irte — me
Molekulare Refraction
η — 1
d
η
2)d
M
ß
V
G
0,1468
0,1473
0,1519
1 G
0,1507
0,1514
0,1561
26,13
26,22
27,03
26,83
26,94
27,78
0,0039
0,0041
0,0042
0,70
0,72
0,75
45,22
45,41
IG
m
{ Cr
G
47,13
R e f r a c t i o n in dem L ö s u n g s z u s t a n d e . Zwei Bestimmungen des
Refractionsäquivalentes in wässeriger Lösung sind mit Lösungen von zwei
verschiedenen Concentrationsstufen ausgeführt worden, von welchen die
stärkere beinahe eine gesättigte Lösung war. Die Resultate sind in der
nächsten Tabelle zusammengestellt; die Molekularrefraction ist immer für
den Strahl G berechnet worden.
M o l e k u l a r e R e f r a c t i o n v o n A m m o n i u m s e l e n a t im L ö s u n g s zustande.
(Je w i c h t
des W a s s e r s :
9,3513
9,5981
Gewicht
des Salzes:
8,7021
7,5040
Proconte
des Salzes
in d e r
Lösung:
48,20
43,88
Dichte
der L ö s u n g
bei
200/40:
1,3627
1,3248
Molekulare
Refraction
des gelösten
Salzes:
Brechungsexponenten
der
Lösung:
Li
G
Na
Tl
F
G
Li
C
Na
Tl
F
G
1,4067
1,4073
1,4098
1,4122
46,84
1,4154
1,4198
1,3990
1,3996
1,4020
1,4043
46,74
1,4073
1,4114
Mittl. Refractionsäquivalent (Gladstone) für den Lösungszustand
46,79
für die Krystalle (Mittel
45,92
aller drei Werte)
Differenz
+
0,87
Optische Vergleichung mit anderen Salzen der Reihe.
Obgleich , die Differenz des Symmetriesystems irgend eine Vergleichung
der Stellungen der respectiven optischen Ellipsoide nicht streng gültig macht,
ist doch die Frage von Interesse, wie die Brechungsexponenten und die
35*
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
Α. Ε. Η. Tutton.
548
molekularen optischen Constanten des selensauren Ammoniums sich zu denjenigen der anderen sieben Salze der Reihe verhalten.
Wenn man die Tabelle der Brechungsexponenten auf Seite 543 mit
derjenigen vergleicht, welche f r ü h e r (diese Zeitschr. 1 8 9 8 , 2 9 , 89) für
die Selenate von Kalium, Rubidium und Cäsium gegeben worden ist, so
sieht man sofort, daß die gesamten Indices des Ammoniumselenats zwischen
denjenigen von Rubidium- und Ciisiumselenat liegen, und beträchtlich n ä h e r
im Durchschnitt an denjenigen des Rubidiumsalzes. Die beste Yergleichung
wird erhalten, wenn man die Mittelwerte (Mittel aller drei Indices α, β und
γ) der Brechungsexponenten f ü r die verschiedenen Salze zusammenstellt,
wie dieses in der nächsten Tabelle für die Wellenlänge des JVa-Lichtes geschehen ist.
Mittelexponenten.
K2S04
1,4952
0,0444
K2Se04
Eb2S04
1,5136
0,0409
Rb2Se04
1,5545
{NH4)2S04
1,5256
0,0438
(NH4)2Se04
1,5694
Cs2S04
1,5635
0,0362
Cs2Se04
1,5997
'
1,5396
®
Die Stellung des Ammoniumsalzes, unmittelbar nach dem Rubidiumsalze, ist also beinahe genau dieselbe in beiden, der Sulfat- und der Selenatreihe. Daher ist die durchschnittliche Refraction des monosymmetrischen
Ammoniumselenats genau so wie zu erwarten war, wenn die Krystalle streng
isomorph mit denjenigen der anderen Glieder der Reihe gewesen wären.
Außerdem steht die Mittelrcfraction des Ammoniumsulfats zu derjenigen
des Ammoniumselenats in ganz ähnlichem Verhältnisse, was diese Eigenschaft
betrifft, wie entsprechende Kalium- und resp. Rubidiumsalze der zwei Reihen
stehen. Dieses wird klar von den in der mittleren Colonne der Tabelle
gegebenen Differenzen gezeigt.
Eine ähnliche Tatsache ist auch mit Hinsicht auf die relativen Dimensionen der optischen Ellipsoide zu bemerken, wenn n u r ihre Axenverhältnisse relativ ausgedrückt w e r d e n , wie es in der nächsten Tabelle der Fall
ist, indem man den Wert für das Kaliumsalz jeder Reihe längs der Axe b
als Einheit annimmt. Obgleich die Richtungen der ellipsoidalen Axen im
Falle des Ammoniumselenats nicht streng vergleichbar mit denjenigen der
anderen Salze sind, ist doch die Stellung des Ammoniumselenats unzweifelh a f t unmittelbar nach dem Rubidiumselenat, genau wie Ammoniumsulfat
dicht nach seinem Analogen, dem Rubidiumsulfat, folgt.
A x e n v e r h ä l t n i s s e des optischen Velocitätsellipsoids.
K2S04
0,9992 : 1
: 0,9975
K2Se04
0,9975 : 1
: 0,9939
Bb2S04
0,9862 : 0,9871 : 0,9869
Rb2Se04
0,9881 : 0,9895 : 0,9853
(.NH4)2S04 0,9742 : 0,9806 : 0,9820
(iVB"4)2Se04 0,9837 : 0,9822 : 0,9688
Cs2SOi
0,9536 : 0 , 9 5 4 7 : 0,9575
Cs2Se04
0,9593 : 0,9602 : 0,9596
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
Aminoniumselenat und die Frage der Isodimorphie in der Alkalisalzreihe.
549
Um eine streng gültige Vergleichung der optischen Constanten zu erhallen, wurde es als notwendig betrachtet, die Werte jener Constanten
für die früher beschriebenen vier Sulfate und drei Selenate neu zu berechnen mit Hilfe der neuen Werte der Dichtigkeiten, welche mittels
der Schwebemethode bestimmt und in der letzten Mitteilung des Verfassers gegeben worden sind (diese Zeitschr. 1905, é l , 386). Die so
erhaltenen Werte sind in den folgenden Tabellen gegeben — sie sind sehr
wahrscheinlich näher an der Wahrheit als die früher gegebenen Werte,
außerdem daß sie streng mit den jetzt mitgeteilten Werte für Ammoniumselenat vergleichbar sind.
S p e c i f i s c h e R e f r a c t i o n ( L o r e n z ) r ^2 , j ! , =
*
' (n + 2)a
Für Strahl C (//«).
[NIl^SO,
BbzSOi
Cs 2 S0 4
(iV7/4)2Se04
K2SeOA
Rb2SeOi
Cs2ScOi
Für Strahl nahe O (H,,).
a
b
c
a
b
e
0,1745
0,1091
0,0831
0,0766
0,1718
0,1088
0,0829
0,0764
0,1712
0,1095
0,0830
0,0759
0,1784
0,1108
0,0846
0,0782
0,1755
0,1106
0,0845
0,0779
0,1749
0,1114
0,0845
0,0774
β0,1473
0,1018
0,0818
0,0765
«0,1468
0,1012
0,0815
0,0763
/0,1519
0,1026
0,0824
0,0765
£0,1514
0,1042
0,0837
0,0784
«0,1507
0,1036
0,0835
0,0782
/0,1561
0,1051
0,0843
0,0783
^ n2 — 1
•1 n2 + 2
M
M o l e k u l a r e R e f r a c t i o n (Lorenz
Für Strali!
K2S04
Eb-ISOI
( m h SO4
Gs.1SOi
K2Se04
Iib.1ScOi
[NH^SEOT
CSISEOI
C(HA).
Für Strahl nahe G (H,.).
a
b
b
e
18,83
21,99
22,54
27,44
e
18,95
22,00
22,46
27,26
a
18,87
22,04
22,89
27,51
19,18
22,44
23,41
28,07
19,14
22,39
23,03
27,99
19,27
22,40
22,95
27,79
22,37
25,52
£26,22
31,05
22,24
25,43
«26,13
30,99
22,56
25,71
γ 27,03
31,03
22,91
26,12
£26,94
31,82
22,77
26,04
«26,83
31,76
23,11
26,30
y 27,78
31,80
Specifische Dispersion n<¡ — ne.
(NII^O,
K2SO4
Rb2SOt
Cs2S04
n.
a
b
0,0039
0,0017
0,0015
0,0016
0,0037
0,0018
0,0016
0,0015
e
0,0037
0,0019
0,0015
0,0015
Molekulare Dispersion mu — m«.;
a
b
0,52
0,31
0,40
0,56
0,49
0,31
0,40
0,55
e
0,49
0,32
0,40
0,53
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
Α. Ε. Η. Tutton.
550
Spccifische Dispersion n 0 — Ho
{NHihSeOi
K2SeOt
Rb2Se04
a
β 0,0041
0,0024
0,0019
b
«0,0039
0,0024
0,0020
Cs2SeOi
0,0019
0,0019
Molekulare Refraction
KvSOt
Rb2SOi
(iV//4)2.S04
Cs2S04
K2Sc04
Rb-1Se04
{NIJ4)2Se04
Cs^ScO^
Vergleichung der
Molekulare Dispersion ntu — ntc.
0
γ 0,0042
0,0025
0,0019
0,0018
a
¡i 0,72
0,54
0,60
b
a 0,70
0,53
0,61
c
y 0,75
0,55
0,59
0,77
0,77
0,77
il
γι
(Gladstone)
M, f ü r S t r a h l C(IIa).
a
32,01
37,61
39,29
47,71
b
31,94
37,52
38,56
47,56
c
32,18
37,53
38,40
47,17
Mittel :
38,46
44,00
£45,41
54,41
38,19
43,82
a 45,22
54,28
38,86
44,36
y 47,13
54,37
38,50
44,06
45,92
54,35
32,04
37,55
38,75
47,48
Refractionsäquiv alente für krystallinische
und gelöste Zustände.
Mittlere
Molekulare Refraction
molekulare
im L ö s u n g s z u s t a n d e ,
Refraction der
SeparatKrystalle :
bestimmungen :
K2SOi
32,04
Rb2S04
37,55
{NH4)2S04
38,75
Cs 2 SOi
47,48
K2Se04
38,50
Rb-tSeOi
44,06
{NH^SeO.,
45,92
Cs 2 Se0 4
54,35
I
1
33,21
33,21
I
I
[
\
l
39,30
39,51
39,47
i
\
J
Í
1
Í
(
39,55
39,71
39,69
39,65
44,72
44,53
46,84
46,74
54,14
54,06
1
Í
1
Í
Mittel:
Dillerenz:
33,21
+1,17
38,21
+0,66
39,43
+0,68
47,27
—0,21
39,65
+1,15
44,63
+0,57
46,79
+0,87
54,10
—0,25
Die bei den Berechnungen angewandten Werte für die specifischen Gewichte, molekularen Gewichte und molekularen Volumina der sieben Salze
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
A m m o n i u m s e l e n a t und die Frage der Isodimorphie in der Alkalisalzrcihe.
551
der Reihe (abgesehen vom Ammoniumselenat) sind diejenigen, welche in der
Mitteilung loc. cit. Seite 386 (1905) gegeben worden sind.
Die drei specifischen und molekularen Constanten des Ammoniumselenats
entsprechen nicht, was ihre Richtungen in dem Krystalle betrifft, den m o r phologischen Axenrichtungen a, b, o der rhombischen Salze, wegen des
verschiedenen Systems (monosymmetrisch) der Krystalle; denn das optische
Ellipsoid ist um die Symmetrieaxe b gedreht.
Sie sind daher markiert
worden, um die Axenrichtung der Indicatrix (Ellipsoid) zu zeigen, und
während « in Wirklichkeit identisch mit der Richtung b ist, entsprechen β
und y nicht den morphologischen Axenrichtungen a und e, sondern anderen
rechtwinkeligen Richtungen in derselben (Symmetrie-) Ebene.
Die Schlußfolgerungen, welche schon in den früheren Mitteilungen des
Verfassers gegeben worden sind, was die Relationen der verschiedenen
Salze und den regulären Einfluß des Atomgewichtes des Alkalimetalles betrifft, mit Hinsicht auf specifische und molekulare Refraction und Dispersion,
sind im Ganzen ebenso gültig f ü r diese neuberechneten Werte.
Die Stellung des Ammoniumselenats in der Selenatreihe ist durchaus
genau dieselbe wie diejenige des Ammoniumsulfats in der Sulfatreihe. W a s
die wichtigste Eigenschaft, die molekulare Refraction, betrifft, ob sie durch
die Formel von L o r e n z oder von G l a d s t o n e u n d D a l e berechnet wird,
so ist diese Stellung immer unmittelbar nach dem Rubidiumsalze. Die Tatsache, daß Ammoniumselenat monosymmetrisch krystallisiert, anstatt r h o m bisch wie die anderen Salze der Reihe, scheint also die specifischen und
molekularen optischen Constanten nicht zu berühren, denn diese sind genau
dieselben, welche von einer streng isomorphen rhombischen Modification
zu erwarten waren. Eine solche rhombische F o r m w ü r d e zweifellos etwas
verschiedene Richtungsdifferenzen zeigen, aber der durchschnittliche W e r t
müßte augenscheinlich derselbe sein, welcher bei der monosymmetrischen
Modification in der Tat beobachtet worden ist. Die Mittelwerte f ü r die
G l a d s t o n e s c h e molekulare Refraction sind in der Tabelle f ü r diese Constanten gegeben u n d werden dieses klar machen.
Ein ähnlicher interessanter Parallelismus zwischen Ammoniumselenat
und Ammoniumsulfat zeigt sich mit Hinsicht auf die Differenz zwischen den
Werten der mittleren molekularen Refraction für den krystallinischen und
den gelösten Zustand.
In beiden Fällen ist die Differenz eine positive,
geringer als diejenige zwischen den zwei Zuständen des Kaliumsalzes, und
nicht weit entfernt von derjenigen zwischen dem krystallinischen und dem
gelösten Rubidiumsalze, während die zwei Zustände des Cäsiumsalzes beider
Reihen eine geringe negative Differenz zeigen. Die Vervollständigung der
beiden Reihen durch die Untersuchung des Ammoniumselenats gibt jetzt
weiteres Gewicht der schon in der Selenalmitteilung (1898) gezogenen Schlußfolgerung, daß die Verallgemeinerung von G l a d s t o n e , wonach »das Re-
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
552
Α. Ε. Η. Tutton.
fractionsäquivalent einer Lösung die Summe der Refractionsäquivalente des
Lösungsmittels und der gelösten Substanzen ist«, im Ganzen eine richtige
ist, denn die geringen durch die Zustandsveränderung veranlaßten Differenzen
liegen auf beiden Seiten der Null und die Ordnung der Differenzen folgt
der Ordnung der Atomgewichte im Falle der Alkalimetalle, während das
Ammoniumsalz die Stellung in der Nähe des Rubidiumsalzes einnimmt,
welche, wie jetzt vom Verfasser gezeigt worden ist, seine eigentliche allgemeine Stellung in der Reihe ist.
Zusammenfassung der Schlußfolgerungen.
Normales Ammoniumselenat, ( N l l ^ S e O n , krystallisiert verschieden von
Ammoniumsulfat und den normalen Sulfaten und Selenaten von Kalium,
Rubidium und Cäsium, nämlich mit monosymmetrischcr Symmetrie, während
die sieben letztgenannten Salze eine streng vergleichbare rhombische Reihe
bilden, deren Charakter und relative Eigenschaften vom Verfasser vollständig in früheren Mitteilungen beschrieben worden sind, und von welchen
die sechs metallischen Salze ein ausgezeichnetes Beispiel der Verallgemeinerung des Verfassers bilden, daß sämtliche krystallographischen Eigenschaften
der Glieder einer solchen isomorphen Reihe Functionen des Atomgewichtes
des Alkalimetalles sind.
Eine rhombische Form des Ammoniumselenates ist einmal (in 1862)
von v. H a u e r erhalten und von v. L a n g beschrieben worden, und
diese historischen Krystalle sind freundlichsterweise von Herrn Prof.
v o n L a n g in Wien dem Verfasser nach London geschickt worden. Ungeachtet dieselben während der zwischenliegenden 44 Jahre durch Zerfließlichkeit etwas gelitten hatten, ist hinreichende und zufriedenstellende Bestätigung der Resultate von v. L a n g erhalten worden, was das Symmetriesystem, die Hauptwinkel und den optischen Charakter betrifft. Es ist aber
durch specifische Gewichtsbestimmungen mit Hilfe der Schwebemethode
gefunden worden, daß die Krystalle wahrscheinlich nicht ganz rein sind,
und zwar daß sie mit etwas Ammoniumsulfat gemischt sind. V. v. L a n g
erwähnte auch in seiner Mitteilung 1862 eine geringe Menge nadelförmiger
Krystalle von abweichendem optischen Charakter, und diese sind identisch
mit der monosymmetrischen Modification des Verfassers gefunden worden.
Diese monosymmetrischen Nadeln erwiesen sich reiner als die rhombischen
Krystalle. und in der Tat scheint praktisch die ganze Verunreinigung in
den rhombischen (isomorphen) Krystallen concentriert zu sein.
Rhombische Mischkrystalle von Ammoniumselenat mit Ammonimsulfat,
Kaliumselenat und resp. Rubidiumselenat sind erhalten worden, und in dem
letzten Falle enthielten sie mehr als 60 Procent von Ammoniumselenat.
Monosymmetrische Mischkrystalle von Ammoniumselenat und Ammoniumsulfat sind ebenfalls erhalten worden, welche 30 Procent des letzteren
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
Ammoniumselenat und die Frage der Isodimorphie in der Alkalisalzreihe.
553
enthalten. Es ist daher angenommen worden, daß Ammoniumselenat dimorph ist, daß die ganze Reihe isodimorph ist, und daß die rhombische
Modification, welche nicht aus der reinen Lösung unter gewöhnlichen Umständen kxystallisiert, in Mischkrystallen existiert.
Die Anwesenheit etwas beigemischten Ammoniumsulfates'in der ν. H a u e r schen Lösung von Ammoniumselenat ist zweifellos die Erklärung der Bildung
der rhombischen Krystalle, welche von v. L a n g gemessen worden sind.
Die monosymmetrischen Krystalle des Ammoniumselenats besitzen eine
sehr ähnliche pseudohexagonale primäre Prismenzone (von beinahe genau
60°), wie diejenigen der rhombischen Krystalle der anderen Salze. In der
Tat kann man die monosymmetrischen Krystalle betrachten, als wenn sie
von den rhombischen deriviert worden sind bloß durch die Neigung der
Basis aus ihrer rechtwinkeligen Stellung in eine schiefe mit Hinsicht auf
das Brachypinakoid. Die Krystalle sind also nach dieser analogen Aufstellung für den Zweck der Beschreibung betrachtet, und die topischen
Parameter sind entsprechenderweise für ein pseudohexagonales Raumgitter
berechnet worden.
Die so erhaltenen topischen Parameter zeigen eine große Ähnlichkeit
mit denjenigen, welche früher für die sieben rhombischen Glieder der Reihe
gegeben worden sind, und ihr Mittelwert liegt zwischen den Mittelwerten
für Rubidium- und Gäsiumselenat.
Das Molekularvolum, die Brechungsexponenten, die Dimensionen des
optischen Ellipsoids und die molekularen optischen Constanten sind auffallend nahe denen, welche zu erwarten wären, wenn die Krystalle wirklich isomorph rhombisch gewesen wären. Mit Hinsicht auf alle diese Constanten nimmt Ammoniumselenat seine Stellung in der Selenatreihe gleich
nach dem Rubidiumselenat ein, genau wie Ammoniumsulfat in seiner Stellung unmittelbar auf Rubidiumsulfat folgt. In der Tat ist es jetzt vom
Verfasser vollkommen bewiesen worden als Resultai der Untersuchungen
sowohl der einfachen Ammoniumsalze als auch der Ammonium enthaltenden
Doppelsalze (diese Zeitschr. 1905, é l , 321), daß die allgemeine krystallographische Stellung des Ammoniums in der Alkalireihe sogleich nach Rubidium folgt.
Als eine secundare Schlußfolgerung ist eine allgemeine Erklärung der
schönen optischen Eigenschaft der Dispersion in gekreuzten Axenebenen
erreicht worden, und die Hauptsätze davon werden in der folgenden
kurzen Mitteilung gegeben.
Brought to you by | University of California
Authenticated
Download Date | 6/4/15 2:58 PM
|
|
https://openalex.org/W1976537999
|
http://old.scielo.br/pdf/bjmbr/v47n9/1414-431X-bjmbr-1414-431X20143869.pdf
|
English
| null |
Effect of physical training on liver expression of activin A and follistatin in a nonalcoholic fatty liver disease model in rats
|
Brazilian Journal of Medical and Biological Research
| 2,014
|
cc-by
| 5,597
|
Brazilian Journal of Medical and Biological Research (2014) 47(9): 746-752, http://dx.doi.org/10.1590/1414-431X20143869
ISSN 1414-431X Brazilian Journal of Medical and Biological Research (2014) 47(9): 746-752, http://dx.doi.org/10.1590/1414-431X20143869
ISSN 1414-431X Brazilian Journal of Medical and Biological Research (2014) 47(9): 746-752, http://dx.doi.org/10.1590/1414-431X20143869
ISSN 1414 431X Abstract Nonalcoholic fatty liver disease (NAFLD) is characterized by fat accumulation in the liver and is associated with obesity and
insulin resistance. Activin A is a member of the transforming growth factor beta (TGF)-b superfamily and inhibits hepatocyte
growth. Follistatin antagonizes the biological actions of activin. Exercise is an important therapeutic strategy to reduce the
metabolic effects of obesity. We evaluated the pattern of activin A and follistatin liver expression in obese rats subjected to
swimming exercise. Control rats (C) and high-fat (HF) diet-fed rats were randomly assigned to a swimming training group (C-
Swim and HF-Swim) or a sedentary group (C-Sed and HF-Sed). Activin bA subunit mRNA expression was significantly higher
in HF-Swim than in HF-Sed rats. Follistatin mRNA expression was significantly lower in C-Swim and HF-Swim than in either
C-Sed or HF-Sed animals. There was no evidence of steatosis or inflammation in C rats. In contrast, in HF animals the severity
of steatosis ranged from grade 1 to grade 3. The extent of liver parenchyma damage was less in HF-Swim animals, with
the severity of steatosis ranging from grade 0 to grade 1. These data showed that exercise may reduce the deleterious effects
of a high-fat diet on the liver, suggesting that the local expression of activin-follistatin may be involved. Key words: Activin A; Follistatin; Steatosis; Nonalcoholic fatty liver disease; Obesity; Exercise R.N. Silva1, P.G. Bueno2, L.R.S. Avo´ 3, K.O. Nonaka2, H.S. Selistre-Arau´ jo2 and A.M.O. Leal3 1Departamento de Fisioterapia, Universidade Federal de Sa˜ o Carlos, Sa˜ o Carlos, SP, Brasil
2Departamento de Cieˆ ncias Fisiolo´ gicas, Universidade Federal de Sa˜ o Carlos, Sa˜ o Carlos, SP, Brasil
3Departamento de Medicina, Universidade Federal de Sa˜ o Carlos, Sa˜ o Carlos, SP, Brasil Correspondence: A.M.O. Leal, Departamento de Medicina, Universidade Federal de Sa˜ o Carlos, Rod. Washington Luı´s, km 235,
13565-905 Sa˜ o Carlos, SP, Brasil. E-mail: angelaleal@ufscar.br
Received January 23, 2014. Accepted May 9, 2014. First published online July 25, 2014. Received January 23, 2014. Accepted May 9, 2014. First published online July 25, 2014. Effect of physical training on liver expression
of activin A and follistatin in a nonalcoholic
fatty liver disease model in rats R.N. Silva1, P.G. Bueno2, L.R.S. Avo´ 3, K.O. Nonaka2, H.S. Selistre-Arau´ jo2 and A.M.O. Leal3 R.N. Silva1, P.G. Bueno2, L.R.S. Avo´ 3, K.O. Nonaka2, H.S. Selistre-Arau´ jo2 and lva1, P.G. Bueno2, L.R.S. Avo´ 3, K.O. Nonaka2, H.S. Selistre-Arau´ jo2 and A.M.O. Lea Introduction The swimming exercise was performed in 30 cm diameter
plastic barrels filled with water to a depth of 50 cm and
maintained at 326-366C. The protocol started with 3 days
of adaptation. On the first and second days of adaptation,
the rats swam for 1 h/day, 30 min at 9:00 am and 30 min
at 5:00 pm without the addition of a weight to the tail. On
the third day of adaptation, the rats swam for 1.5 h/day,
45 min at 9:00 am and 45 min at 5:00 pm again without
the addition of a weight. On the fourth day, an external
weight corresponding to 1% of the animal’s body mass
was attached to the base of the tail, and rats swam for
90 min/day. During the following weeks, rats swam for
90 min/day with the tail-weight increased weekly until
reaching 3% and then 5% of the animal’s body mass (14-
16). All animals were towel dried before being returned to
their cages. The sedentary groups were placed in shallow
water for 5 min, towel dried and then returned to their
cages at the same time as the exercised groups. Both the
control rats and the HF diet rats were able to carry out the
exercise protocol similarly. reflecting increased activin A bioactivity. Activin A
contributes to the pathogenesis of NAFLD by inhibiting
insulin sensitivity and metabolism of glucose and fatty
acids. It has been demonstrated that activin A promotes
collagen 3 and TGF-b1 expression and matrix metallo-
proteinase (MMP) activity, suggesting that activin A may
promote hepatic fibrosis. On the other hand, activin A
induces mitochondrial b-oxidation and downregulates de
novo fatty acid synthesis, suggesting that activin A may
reduce lipid accumulation in hepatocytes (4,6). A recent review of the relationship between exercise
and the liver and its implications for the therapy of fatty
liver disorders concluded that the effect of exercise on
liver fat reduction compared favorably with most of the
available pharmacological therapies (8). Physical exercise
is linked to improved glucose homeostasis and enhanced
insulin sensitivity, and is a basic means for the prevention
of type 2 diabetes and the management of both type 1 and
type 2 diabetes (9). Physical exercise has also been
shown to have positive effects in the prevention and
attenuation of hepatic steatosis (10). After the training period, i.e., 48 h after the last
exercise session, rats were sacrificed by decapitation. Introduction Nonalcoholic fatty liver disease (NAFLD) is the most
common type of chronic liver disorder and includes a
broad spectrum of liver damage, ranging from simple
steatosis to steatohepatitis (NASH) and cirrhosis (1,2). Evaluation of these histopathologic characteristics is
possible through a liver biopsy (1,2). NAFLD is closely
associated with obesity and increased lipolysis due to loss
of insulin sensitivity and impaired antilipolytic action of
insulin, the main factor responsible for the development of
hepatic steatosis (2). by inhibiting the replication of hepatocytes and inducing
apoptosis (3). The biological actions of activin are
counteracted by follistatin, an unrelated glycoprotein that
binds mature dimeric activin A with high affinity and blocks
activin signaling (3,6). Follistatin is associated with liver
regeneration after partial hepatectomy (7). Activin signal transduction occurs through transmem-
brane serine/threonine kinase receptors. Activin binds to
the activin type II receptors (ActRII), transphosphorylates
type I receptors (activin receptor-like kinases, ALKs) (1-7)
and forms the ActRII-IB/(ALK) 4 complex. This activated
activin receptor complex recruits the receptor-associated
Smads, which transmit the activin signals to the cell
nucleus where they regulate the transcription of target
genes (3,5,6). Activins, members of the transforming growth factor
beta (TGF-b) superfamily, are bioactive dimeric proteins
composed of two beta subunits. Activin A, a homodimer
composed of two beta A subunits, is involved in the
pathogenesis of a variety of inflammatory diseases
ranging from rheumatoid arthritis to inflammatory bowel
disease and atherosclerosis, and also in several liver
disorders, including NAFLD and liver fibrosis (3-5). Activin
A is considered to be a negative regulator of liver growth Activin A seems to have multiple roles in NAFLD. It
has been demonstrated that serum levels of activin A
and follistatin and the activin A/follistatin mRNA ratio in
liver are increased in patients with NAFLD, potentially www.bjournal.com.br Braz J Med Biol Res 47(9) 2014 Activin and follistatin in NAFLD and exercise 747 reflecting increased activin A bioactivity. Activin A
contributes to the pathogenesis of NAFLD by inhibiting
insulin sensitivity and metabolism of glucose and fatty
acids. It has been demonstrated that activin A promotes
collagen 3 and TGF-b1 expression and matrix metallo-
proteinase (MMP) activity, suggesting that activin A may
promote hepatic fibrosis. On the other hand, activin A
induces mitochondrial b-oxidation and downregulates de
novo fatty acid synthesis, suggesting that activin A may
reduce lipid accumulation in hepatocytes (4,6). Glucose and insulin tolerance tests Glucose tolerance tests (GTTs) were performed after
the acclimatization period, at 3, 4, 8, and 12 weeks of
dietary treatment, and after exercise training by the 16th
week. After an overnight fast, unanesthetized rats were
injected intraperitoneally (ip) with 50% glucose solution,
1.5 g/kg body weight (BW). Blood samples were obtained
from the tail vein before and at 30, 60, 90, and 120 min
after glucose injection. Insulin tolerance tests (ITTs) were
performed after the acclimatization period and at 8 and 12
weeks of dietary treatment. After an overnight fast,
unanesthetized rats were injected ip with Novolin R
human insulin (Novo Nordisk, Brazil), 0.75 U/kg BW. Blood samples were obtained from the tail vein before
and at 15, 30, 60, and 90 min after the insulin challenge. Blood glucose concentrations were measured with an
Accu-Check glucose meter (Roche Diagnostic, USA). We hypothesized that activin A/follistatin may partici-
pate in the beneficial effect of exercise on NAFLD, which
still has not been elucidated. In the present study, we
evaluated the patterns of activin A and follistatin expres-
sion in the livers of obese rats subjected to swimming
exercise. Histological analysis Formalin-fixed and paraffin-embedded liver sections
were stained with hematoxylin and eosin (HE) and
Masson’s trichrome for histopathological examination. The severity of steatosis was graded by the extent of
parenchymal involvement. Grade 0, ,5% of the parench-
yma was involved; grade 1, 5 to 33%; grade 2, 34 to 66%;
grade 3, .66%. Other parameters assessed were the
presence or absence of inflammation and fibrosis score
(17). Animals and experimental groups All procedures were approved by the Ethics Committee
of Universidade Federal de Sa˜ o Carlos (#048/2007). Male
Wistar rats (190±18.4 g) were acclimatized in individual
cages under controlled temperature, humidity and lighting
(12-h dark/light cycle) conditions, and with free access to
water and standard rodent chow. After 7 days, animals
were randomly assigned to two groups: control rats (C,
n=15) fed standard rodent chow, and high-fat diet-fed rats
(HF, n=15), fed 60% kcal as fat (Research Diets, USA). Animals and food intake were weighed daily. After 12
weeks, animals in both groups were randomly divided into
two groups, a swimming training group (C-Swim and
HF-Swim) and a sedentary group (C-Sed and HF-Sed) with
6-7 rats/group. www.bjournal.com.br Introduction Blood plasma was collected and stored at ––206C; the liver
was dissected, weighed, frozen immediately in liquid
nitrogen, and stored at ––806C until analysis. Studies in diabetic and obese rats have shown that
exercise can modulate follistatin in muscle and fat tissue
(11,12). A recent study also demonstrated that plasma
follistatin increased in response to exercise, most likely
originating from the liver (13). However, there is no
evidence of a relationship between exercise and activin. We hypothesized that activin A/follistatin may partici-
pate in the beneficial effect of exercise on NAFLD, which
still has not been elucidated. In the present study, we
evaluated the patterns of activin A and follistatin expres-
sion in the livers of obese rats subjected to swimming
exercise. Studies in diabetic and obese rats have shown that
exercise can modulate follistatin in muscle and fat tissue
(11,12). A recent study also demonstrated that plasma
follistatin increased in response to exercise, most likely
originating from the liver (13). However, there is no
evidence of a relationship between exercise and activin. Exercise protocol Rats in the exercise groups (C-Swim and HF-Swim)
were exercised by swimming 1.5 h/day, 45 min at 9:00
am and 45 min at 5:00 pm, 5 days per week, for 4 weeks. )
The histological slides were analyzed and photo-
graphed with an Olympus1 BX 51 optical microscope Braz J Med Biol Res 47(9) 2014 748 R.N. Silva et al. Figure 1. Body weight (g) of control and high-fat
diet-fed rats. Data are reported as means±SE
for n=6-7 rats/group. *P,0.05, sedentary high-
fat diet group (HF-Sed) vs sedentary control
group (C-Sed);
{P,0.05, C-Sed vs exercised
control group (C-Swim);
{P,0.05, HF-Sed vs
exercised high-fat diet group (HF-Swim);
1P,0.05, C-Sed 12th week vs 16th week;
"P,0.05, HF-Sed 12th week vs 16th week;
**P,0.05, HF-swim 12th week (pre-exercise) vs
16th week (post-exercise) (t-test). Figure 1. Body weight (g) of control and high-fat
diet-fed rats. Data are reported as means±SE
for n=6-7 rats/group. *P,0.05, sedentary high-
fat diet group (HF-Sed) vs sedentary control
group (C-Sed);
{P,0.05, C-Sed vs exercised
control group (C-Swim);
{P,0.05, HF-Sed vs
exercised high-fat diet group (HF-Swim);
1P,0.05, C-Sed 12th week vs 16th week;
"P,0.05, HF-Sed 12th week vs 16th week;
**P,0.05, HF-swim 12th week (pre-exercise) vs
16th week (post-exercise) (t-test). Figure 1. Body weight (g) of control and high-fat
diet-fed rats. Data are reported as means±SE
for n=6-7 rats/group. *P,0.05, sedentary high-
fat diet group (HF-Sed) vs sedentary control
group (C-Sed);
{P,0.05, C-Sed vs exercised
control group (C-Swim);
{P,0.05, HF-Sed vs
exercised high-fat diet group (HF-Swim);
1P,0.05, C-Sed 12th week vs 16th week;
"P,0.05, HF-Sed 12th week vs 16th week;
**P,0.05, HF-swim 12th week (pre-exercise) vs
16th week (post-exercise) (t-test). (Olympus Corporation, Japan). The pathologist who
evaluated the sections and performed the histological
assessments was unaware of the treatment groups. GCTGA-39) (18,19). In addition, a rat liver cDNA standard
was run in duplicate for every plate to produce a standard
curve for quantification. The relative amounts of RNAs
were calculated using the comparative Ct method. The
expression of all genes was normalized to glyceralde-
hyde-3-phosphate dehydrogenase (GAPDH) expression. Body weight The body weight of HF diet-fed animals was sig-
nificantly higher than that of control animals from week 6
until the end of the training period (Figure 1). At week 16,
exercised animals (both control and HF diet-fed) had
significantly lower body weights than sedentary animals
(both controls and HF diet-fed). In HF diet-fed animals,
post-exercise body weight at week 16 was significantly
lower than pre-exercise body weight at week 12. Exercise
training was thus able to prevent physiological weight gain
associated with the standard rodent chow. RNA extraction and quantitative real-time PCR q
Frozen liver samples (40 mg) were homogenized on
ice in TRIzol reagent (Invitrogen Corporation, USA)
following the manufacturer’s instructions. The purity and
concentration of total RNA were determined by measuring
the absorbance of aliquots at a ratio of 260/280 nm. The
integrity of RNA was checked on 2% agarose gels by
inspecting the electrophoretic pattern of 28S and 18S
ribosomal RNA. Total RNA (1 mg) from each sample was
treated with DNase I (Invitrogen Corporation) to remove
contaminating genomic DNA and reverse transcribed to
synthesize cDNA using Moloney murine leukemia virus
reverse transcriptase (Promega Corporation, USA). For
real-time PCR, 20 ng cDNA and 0.4 mM of each primer
were used in a 25 mL reaction volume containing SYBR
Green PCR Master Mix (Applied Biosystems Inc., USA). The primer sequences used were: beta-A activin (forward
primer: 59-ATGGACCTAACTCTCAGCCAGA-39; reverse
primer: 59-CTCTCCCCCTTCAAGCCCAT-39); follistatin
(forward primer: 59-GGCGTACTGCTTGAAGTGAA-39;
reverse primer: 59-GGGAAGCTGTAGTCCTGGTC-39);
GAPDH (forward primer: 59-GATGCTGGTGCTGAGTAT
GTCG-39; reverse primer: 59-GTGGTGCAGGATGCATT Biochemical analysis Alanine transaminase (ALT), aspartate transaminase
(AST), gamma-glutamyl transaminase (GGT), and
plasma lipids, including total cholesterol and triglycerides,
were measured enzymatically with commercially available
assay kits (Katal Biotecnolo´ gica, Brazil). Statistical analysis Statistical analyses were performed by paired and
unpaired Student t-tests and analysis of variance
(ANOVA). The Tukey multiple comparisons test was used
for post hoc analysis of between-group comparisons. Data are reported as means±SE. P,0.05 was consid-
ered to be statistically significant. Data analyses were
carried out using the STATISTICA data analysis software
system, version 7 (StatSoft Inc., USA). Braz J Med Biol Res 47(9) 2014 Liver expression of mRNA Activin A subunit mRNA expression was significantly
higher in HF-Swim than in HF-Sed animals, but no
difference was observed between C-Sed and C-Swim
animals. Follistatin mRNA was significantly lower in
C-Swim and HF-Swim animals than in either C-Sed or
HF-Sed animals (Figure 3). Liver morphology Liver morphology varied significantly among groups. There was no evidence of steatosis or inflammation
(grade 0) in rats fed standard rodent chow. In contrast, the
severity of steatosis ranged from grade 1 to grade 3 in rats
fed the HF diet. This was not associated with inflammation
or fibrosis. After the training period, rats fed the HF diet
showed improvement in the extent of damage to the liver
parenchyma, with severity of steatosis ranging from grade
0 to grade 1. Representative images of liver sections are
shown in Figure 4. GTT and ITT tests Rats fed an HF diet had significantly higher glycemia
in response to both glucose and insulin injections than Braz J Med Biol Res 47(9) 2014 www.bjournal.com.br Activin and follistatin in NAFLD and exercise 749 t
l
t
ft
12
k
f th
di t
i
(Fi
2A
Figure 2. Glucose and insulin tolerance tests. Analysis of
glycemia profile by the A, glucose tolerance test (GTT) and the
B, insulin tolerance test (ITT) at the end of the 12th week in rats
fed a high-fat diet. C, Analysis of glycemia profile by the GTT
pre-exercise (12th week) and post-exercise (16th week) training. Data are reported as means±SE for n=15 (C and HF groups)
and n=6-7 rats/subgroup. *P,0.05, high-fat diet group (HF) vs
control group (C). {P,0.05, exercised high-fat diet group pre-
exercise (HF-Pre-ex) vs exercised control group pre-exercise
(C-Pre-ex); {P,0.05, HF-Pre-ex vs exercised high-fat diet group
post-exercise (HF-Post-ex) (t-test). response in HF-Swim rats (Figure 2C). No difference was
observed between pre- and post-exercise glycemia in
response to glucose injection in C-Swim rats. Liver enzymes and lipids ALT levels were significantly higher in HF-Sed animals
than in C-Sed animals (41.7±2.7 vs 26.9±1.4 U/L,
P,0.05) and significantly lower in HF-Swim animals than
in HF-Sed animals (31.6±2.1 vs 41.7±2.7 U/L, P,0.05). Plasma AST, GGT, total cholesterol and triglycerides
levels did not vary among groups (data not shown). www.bjournal.com.br Discussion The present data showed that exercise reduced the
deleterious effects of an HF diet on the liver and altered
both hepatic activin beta A subunit and follistatin mRNA
expression in this rat model of NAFLD. p
Decreased glucose tolerance has previously been
shown to accompany obesity induced by an HF diet (20). Here, exercise training reduced insulin resistance in rats
fed an HF diet. It is well established that exercise is linked
to enhanced insulin sensitivity in diet-induced obese rats
by activation of components of the insulin-signaling
cascade pathway in both skeletal muscle and liver
(21,22). However, it is important to consider the intensity,
frequency and duration of the exercise. It has been
reported that exercise training was able to improve insulin
activity in fat tissue, and that a stronger effect was seen
with high- than with moderate-intensity exercise (23,24). In this study, we also demonstrated that moderate
aerobic exercise reduces body weight and attenuates
hepatic steatosis. Previous studies have reported that
moderate aerobic exercise promotes a reduction of
body weight and adiposity, improves blood lipid profiles
and attenuates hepatic steatosis, suppressing HF Decreased glucose tolerance has previously been
shown to accompany obesity induced by an HF diet (20). Here, exercise training reduced insulin resistance in rats
fed an HF diet. It is well established that exercise is linked
to enhanced insulin sensitivity in diet-induced obese rats
by activation of components of the insulin-signaling
cascade pathway in both skeletal muscle and liver
(21,22). However, it is important to consider the intensity,
frequency and duration of the exercise. It has been
reported that exercise training was able to improve insulin
activity in fat tissue, and that a stronger effect was seen
with high- than with moderate-intensity exercise (23,24). Figure 2. Glucose and insulin tolerance tests. Analysis of
glycemia profile by the A, glucose tolerance test (GTT) and the
B, insulin tolerance test (ITT) at the end of the 12th week in rats
fed a high-fat diet. C, Analysis of glycemia profile by the GTT
pre-exercise (12th week) and post-exercise (16th week) training. Data are reported as means±SE for n=15 (C and HF groups)
and n=6-7 rats/subgroup. *P,0.05, high-fat diet group (HF) vs
control group (C). {P,0.05, exercised high-fat diet group pre-
exercise (HF-Pre-ex) vs exercised control group pre-exercise
(C-Pre-ex); {P,0.05, HF-Pre-ex vs exercised high-fat diet group
post-exercise (HF-Post-ex) (t-test). Discussion In this study, we also demonstrated that moderate
aerobic exercise reduces body weight and attenuates
hepatic steatosis. Previous studies have reported that
moderate aerobic exercise promotes a reduction of
body weight and adiposity, improves blood lipid profiles
and attenuates hepatic steatosis, suppressing HF control rats after 12 weeks of the diet regimen (Figure 2A
and B). The post-exercise glycemic response to glucose
injection was significantly lower than the pre-exercise test Braz J Med Biol Res 47(9) 2014 750 R.N. Silva et al. Figure 3. Liver expression mRNA. A, Activin bA
and B, follistatin mRNA expression in liver of
Wistar rats. Data are reported as means±SE for
n=6-7 rats/group. {P,0.05, sedentary control
group (C-Sed) vs exercised control group (C-
Swim);
{P,0.05, sedentary high-fat diet group
(HF-Sed) vs exercised high-fat diet group (HF-
Swim) (t-test). Figure 3. Liver expression mRNA. A, Activin bA
and B, follistatin mRNA expression in liver of
Wistar rats. Data are reported as means±SE for
n=6-7 rats/group. {P,0.05, sedentary control
group (C-Sed) vs exercised control group (C-
Swim);
{P,0.05, sedentary high-fat diet group
(HF-Sed) vs exercised high-fat diet group (HF-
Swim) (t-test). fatty acid synthesis/uptake and oxidation/export (6,30). diet-induced steatosis (10,25,26). It has long been ob-
served that weight loss by caloric restriction alone or in
combination with exercise prevents NAFLD (27). However, there is increasing evidence that exercise per
se beneficially reduces hepatic steatosis independent of
weight loss (28) and that exercise may offer additional
benefits related to hepatic mitochondrial function com-
pared to a restricted diet alone in rodents (29). The results showed that swimming training minimized
the liver tissue damage caused by an HF diet as
previously demonstrated (10). The beneficial effects of
exercise have been attributed to increased insulin
sensitivity and fat oxidation and decreased lipid synthesis
by the liver through activation of the AMP-activated
protein kinase (AMPK) pathway (10,32). However,
mechanistically, the effect of exercise on the pathogen-
esis of NAFLD is not understood completely. NAFLD comprises a continuum of liver diseases
ranging from simple fat deposition (steatosis) to inflam-
mation, extensive fibrosis and cirrhosis that develop in the
absence of excessive alcohol intake (1). NAFLD is
associated with obesity, insulin resistance and the
metabolic syndrome (30). In this study, we used a
previously described rodent model of NAFLD (11,31)
and a validated histological scoring system to describe the
histologic findings in this model (17). Figure 3. Liver expression mRNA. A, Activin bA
and B, follistatin mRNA expression in liver of
Wistar rats. Data are reported as means±SE for
n=6-7 rats/group.
{P,0.05, sedentary control
group (C-Sed) vs exercised control group (C-
Swim);
{P,0.05, sedentary high-fat diet group
(HF-Sed) vs exercised high-fat diet group (HF-
Swim) (t-test). References 1. Brunt EM. Nonalcoholic steatohepatitis: definition and
pathology. Semin Liver Dis 2001; 21: 3-16, doi: 10.1055/s-
2001-12925. 10. Schultz A, Mendonca LS, Aguila MB, Mandarim-de-Lacerda
CA. Swimming training beneficial effects in a mice model of
nonalcoholic fatty liver disease. Exp Toxicol Pathol 2012;
64: 273-282, doi: 10.1016/j.etp.2010.08.019. 2. Smith BW, Adams LA. Non-alcoholic fatty liver disease. Crit Rev Clin Lab Sci 2011; 48: 97-113, doi: 10.3109/
10408363.2011.596521. 11. Bueno PG, Bassi D, Contrera DG, Carnielli HM, Silva RN,
Nonaka KO, et al. Post-exercise changes in myostatin and
actRIIB expression in obese insulin-resistant rats. Mol Cell
Endocrinol 2011; 339: 159-164, doi: 10.1016/j.mce. 2011.04.006. 3. Rodgarkia-Dara C, Vejda S, Erlach N, Losert A, Bursch W,
Berger W, et al. The activin axis in liver biology and disease. Mutat
Res
2006;
613:
123-137,
doi:
10.1016/
j.mrrev.2006.07.002. 12. Dutra DB, Bueno PG, Silva RN, Nakahara NH, Selistre-
Araujo HS, Nonaka KO, et al. Expression of myostatin,
myostatin receptors and follistatin in diabetic rats submitted
to exercise. Clin Exp Pharmacol Physiol 2012; 39: 417-422,
doi: 10.1111/j.1440-1681.2012.05690.x. 4. Yndestad A, Haukeland JW, Dahl TB, Bjoro K, Gladhaug IP,
Berge C, et al. A complex role of activin A in non-alcoholic
fatty liver disease. Am J Gastroenterol 2009; 104: 2196-
2205, doi: 10.1038/ajg.2009.318. 5. Hashimoto O, Funaba M. Activin in glucose metabolism. Vitam Horm 2011; 85: 217-234, doi: 10.1016/B978-0-12-
385961-7.00011-1. 13. Hansen J, Brandt C, Nielsen AR, Hojman P, Whitham M,
Febbraio MA, et al. Exercise induces a marked increase in
plasma follistatin: evidence that follistatin is a contraction-
induced hepatokine. Endocrinology 2011; 152: 164-171,
doi: 10.1210/en.2010-0868. 6. Yndestad A, Haukeland JW, Dahl TB, Halvorsen B, Aukrust
P. Activin A in nonalcoholic fatty liver disease. Vitam Horm
2011; 85: 323-342, doi: 10.1016/B978-0-12-385961-7. 00015-9. 14. Matsakas A, Bozzo C, Cacciani N, Caliaro F, Reggiani C,
Mascarello F, et al. Effect of swimming on myostatin
expression in white and red gastrocnemius muscle and in
cardiac muscle of rats. Exp Physiol 2006; 91: 983-994, doi:
10.1113/expphysiol.2006.033571. 7. Ooe H, Chen Q, Kon J, Sasaki K, Miyoshi H, Ichinohe N,
et al. Proliferation of rat small hepatocytes requires follistatin
expression. J Cell Physiol 2012; 227: 2363-2370, doi:
10.1002/jcp.22971. 15. Leme JA, Gomes RJ, de Mello MA, Luciano E. Effects of
short-term physical training on the liver IGF-I in diabetic
rats. Growth Factors 2007; 25: 9-14, doi: 10.1080/
08977190701210693. 8. Johnson NA, Keating SE, George J. Discussion The results showed
that an HF diet produced different degrees of fat
accumulation but no inflammation or fibrosis. NAFLD is
a complex disorder whose main initial pathological
process is the accumulation of fat within the hepatocytes. It is believed to result from an imbalance between hepatic We showed for the first time that activin and follistatin
may be involved in the beneficial effects of exercise on
NAFLD. We observed that the expression of activin A and
follistatin mRNAs varied reciprocally in exercised and
sedentary HF-fed animals, suggesting a local adaptive
regulation promoting an increase in the biological effect of
activin A. Activin A has been previously reported to be
profoundly involved in liver function and in the pathogen-
esis of liver disease, including NAFLD (3,4,6). Follistatin is
a binding protein that acts as an extracellular factor to
bind and inactivate activin (33). It has recently been
demonstrated that in mice with global inactivating Braz J Med Biol Res 47(9) 2014 Figure 4. Histological analysis of liver from rats
fed standard rodent chow and a high-fat diet. Representative microphotographs of hematox-
ylin-eosin-stained sections (bar=20 mm) are
shown. Red arrows indicate fat accumulation. C-Sed: sedentary control group; C-Swim: exer-
cised control group; HF-Sed: sedentary high-fat
diet group; HF-Swim: exercised high-fat diet
group. Braz J Med Biol Res 47(9) 2014
www.bjournal.com.br Figure 4. Histological analysis of liver from rats
fed standard rodent chow and a high-fat diet. Representative microphotographs of hematox-
ylin-eosin-stained sections (bar=20 mm) are
shown. Red arrows indicate fat accumulation. C-Sed: sedentary control group; C-Swim: exer-
cised control group; HF-Sed: sedentary high-fat
diet group; HF-Swim: exercised high-fat diet
group. Braz J Med Biol Res 47(9) 2014 www.bjournal.com.br Activin and follistatin in NAFLD and exercise 751 In spite of their novelty, the data presented here
should be considered along with the study limitations. The
modulation of activin and follistatin expression by exercise
was shown only at the transcriptional level, thus post-
transcriptional regulation cannot be ruled out. In addition,
the mechanisms involved in the action of activin/follistatin
on hepatic steatosis were not investigated in the present
study. However, the findings may offer insight into the
development of pharmacological strategies contributing to
the control of obesity comorbidities such as NAFLD. Discussion mutations that deplete the circulating activin antagonists
follistatin-like-3 or the follistatin 315 isoform, insulin
signaling in liver was enhanced, and paradoxically,
hepatic steatosis developed and phosphoenolpyruvate
carboxykinase (PEPCK) expression was not suppressed,
suggesting dual actions of activin in hepatocytes (34). gg
g
p
y
(
)
The activin/follistatin circuitry is complex, with local
autocrine/paracrine functions, and at the same time may
be self-modulated (35,36). Many studies have described
potential mechanisms to explain the role of activin/
follistatin in exercise-induced liver adaptation (4,5). In this
regard, activins have been shown to be associated with
insulin production and insulin sensitivity, as well as with fat
oxidation in liver and other tissues (37). In addition, activin
A can act as an anti-inflammatory cytokine (38), and
inflammation is an important mechanism involved in both
insulin resistance and NAFLD (5,39). Moreover, it has
recently been shown that exercise training reduces
hepatic inflammation, injury and fibrosis by suppressing
macrophage infiltration in diet-induced obesity in mice
(40). In conclusion, the results showed that exercise may
reduce the deleterious effects of an HF diet on the liver
and suggest that the local expression of activin and
follistatin may be involved. Further studies are necessary
to investigate the mechanisms of regulation of the activin/
follistatin circuitry in metabolic and liver diseases. Acknowledgments Research supported by FAPESP (#2007/54854-0)
and CNPq. References Exercise and the liver:
implications for therapy in fatty liver disorders. Semin Liver
Dis 2012; 32: 65-79, doi: 10.1055/s-0032-1306427. 16. Voltarelli FA, Gobatto CA, de Mello MA. Determination of
anaerobic threshold in rats using the lactate minimum test. Braz J Med Biol Res 2002; 35: 1389-1394, doi: 10.1590/
S0100-879X2002001100018. 9. Colberg SR, Albright AL, Blissmer BJ, Braun B, Chasan-
Taber L, Fernhall B, et al. Exercise and type 2 diabetes:
American College of Sports Medicine and the American
Diabetes Association: joint position statement. Exercise and
type 2 diabetes. Med Sci Sports Exerc 2010; 42: 2282-2303,
doi: 10.1249/MSS.0b013e3181eeb61c. 17. Kleiner DE, Brunt EM, Van Natta M, Behling C, Contos MJ,
Cummings OW, et al. Design and validation of a histological Braz J Med Biol Res 47(9) 2014 www.bjournal.com.br 752 R.N. Silva et al. scoring system for nonalcoholic fatty liver disease. Hepatology 2005; 41: 1313-1321, doi: 10.1002/hep.20701. restriction for prevention of nonalcoholic fatty liver disease
in the OLETF rat model. Am J Physiol Gastrointest Liver
Physiol 2011; 300: G874-G883, doi: 10.1152/ajpgi. 00510.2010. 18. 18. Gold EJ, Francis RJ, Zimmermann A, Mellor SL, Cranfield
M, Risbridger GP, et al. Changes in activin and activin
receptor subunit expression in rat liver during the develop-
ment of CCl4-induced cirrhosis. Mol Cell Endocrinol 2003;
201: 143-153, doi: 10.1016/S0303-7207(02)00417-3. 30. 30. Utzschneider KM, Kahn SE. Review: The role of insulin
resistance in nonalcoholic fatty liver disease. J Clin
Endocrinol Metab 2006; 91: 4753-4761, doi: 10.1210/jc. 2006-0587. 19. Ferreira MC, Cavallo IK, Florio P, Petraglia F, Reis FM. Activin betaA subunit, follistatin and follistatin-like 3 are
expressed in the endometrium of ovariectomized rats and
regulated by estrogen replacement. J Mol Histol 2008; 39:
535-541, doi: 10.1007/s10735-008-9194-x. 31. London RM, George J. Pathogenesis of NASH: animal
models. Clin Liver Dis 2007; 11: 55-74, viii, doi: 10.1016/
j.cld.2007.02.010. 32. Park H, Kaushik VK, Constant S, Prentki M, Przybytkowski
E, Ruderman NB, et al. Coordinate regulation of malonyl-
CoA decarboxylase, sn-glycerol-3-phosphate acyltransfer-
ase, and acetyl-CoA carboxylase by AMP-activated protein
kinase in rat tissues in response to exercise. J Biol Chem
2002; 277: 32571-32577, doi: 10.1074/jbc.M201692200. 20. Woods SC, Seeley RJ, Rushing PA, D’Alessio D, Tso P. A
controlled high-fat diet induces an obese syndrome in rats. J Nutr 2003; 133: 1081-1087. 21. Yaspelkis BB, III, Lessard SJ, Reeder DW, Limon JJ, Saito
M, Rivas DA, et al. References Exercise reverses high-fat diet-induced
impairments on compartmentalization and activation of
components of the insulin-signaling cascade in skeletal
muscle. Am J Physiol Endocrinol Metab 2007; 293: E941-
E949, doi: 10.1152/ajpendo.00230.2007. 33. Shimonaka M, Inouye S, Shimasaki S, Ling N. Follistatin
binds to both activin and inhibin through the common
subunit. Endocrinology 1991; 128: 3313-3315, doi: 10.1210/
endo-128-6-3313. 34. Ungerleider NA, Bonomi LM, Brown ML, Schneyer AL. Increased activin bioavailability enhances hepatic insulin
sensitivity while inducing hepatic steatosis in male mice. Endocrinology 2013; 154: 2025-2033, doi: 10.1210/en. 2012-1844. 22. De Souza CT, Frederico MJ, da Luz G, Cintra DE, Ropelle
ER, Pauli JR, et al. Acute exercise reduces hepatic glucose
production through inhibition of the Foxo1/HNF-4alpha
pathway in insulin resistant mice. J Physiol 2010; 588:
2239-2253, doi: 10.1113/jphysiol.2009.183996. 35. Leal AM, Blount AL, Donaldson CJ, Bilezikjian LM, Vale
WW. Regulation of follicle-stimulating hormone secre-
tion by the interactions of activin-A, dexamethasone and
testosterone in anterior pituitary cell cultures of male
rats. Neuroendocrinology 2003; 77: 298-304, doi: 10.1159/
000070896. 23. Wisloff U, Najjar SM, Ellingsen O, Haram PM, Swoap S, Al-
Share Q, et al. Cardiovascular risk factors emerge after
artificial selection for low aerobic capacity. Science 2005;
307: 418-420, doi: 10.1126/science.1108177. 24. Wisloff U, Najjar SM, Ellingsen O, Haram PM, Swoap S, Al-
Share Q, et al. Cardiovascular risk factors emerge after
artificial selection for low aerobic capacity. Science 2005;
307: 418-420, doi: 10.1126/science.1108177. 36. Bilezikjian LM, Blount AL, Leal AM, Donaldson CJ, Fischer
WH, Vale WW. Autocrine/paracrine regulation of pituitary
function by activin, inhibin and follistatin. Mol Cell Endocrinol
2004; 225: 29-36, doi: 10.1016/j.mce.2004.02.010. 25. Estadella D, Oyama LM, Damaso AR, Ribeiro EB, Oller Do
Nascimento CM. Effect of palatable hyperlipidic diet on lipid
metabolism of sedentary and exercised rats. Nutrition 2004;
20: 218-224, doi: 10.1016/j.nut.2003.10.008. 37. Mine T, Yasuda H, Fujita T, Hasegawa Y. Effects of inhibin
on activin A-Induced glucose metabolism in rat hepatocytes. Endocrine 1996; 5: 41-46, doi: 10.1007/BF02738654. 26. Marques CM, Motta VF, Torres TS, Aguila MB, Mandarim-
de-Lacerda CA. Beneficial effects of exercise training (tread-
mill) on insulin resistance and nonalcoholic fatty liver disease
in high-fat fed C57BL/6 mice. Braz J Med Biol Res 2010; 43:
467-475, doi: 10.1590/S0100-879X2010007500030. 38. Jones KL, Mansell A, Patella S, Scott BJ, Hedger MP, de
Kretser DM, et al. Activin A is a critical component of the
inflammatory response, and its binding protein, follistatin,
reduces mortality in endotoxemia. References Proc Natl Acad Sci U S A
2007; 104: 16239-16244, doi: 10.1073/pnas.0705971104. 27. Ueno T, Sugawara H, Sujaku K, Hashimoto O, Tsuji R,
Tamaki S, et al. Therapeutic effects of restricted diet and
exercise in obese patients with fatty liver. J Hepatol 1997;
27: 103-107, doi: 10.1016/S0168-8278(97)80287-5. 39. Schenk S, Saberi M, Olefsky JM. Insulin sensitivity:
modulation by nutrients and inflammation. J Clin Invest
2008; 118: 2992-3002, doi: 10.1172/JCI34260. 28. Johnson NA, George J. Fitness versus fatness: moving
beyond weight loss in nonalcoholic fatty liver disease. Hepatology 2010; 52: 370-381, doi: 10.1002/hep.23711. 40. Kawanishi N, Yano H, Mizokami T, Takahashi M, Oyanagi
E, Suzuki K. Exercise training attenuates hepatic inflamma-
tion, fibrosis and macrophage infiltration during diet
induced-obesity in mice. Brain Behav Immun 2012; 26:
931-941, doi: 10.1016/j.bbi.2012.04.006. 29. Rector RS, Uptergrove GM, Morris EM, Borengasser SJ,
Laughlin MH, Booth FW, et al. Daily exercise vs caloric Braz J Med Biol Res 47(9) 2014 www.bjournal.com.br
|
https://openalex.org/W4387349785
|
https://journals.udau.cherkasy.ua/index.php/visnyk/article/download/44/39
|
Ukrainian
| null |
CROP FORMATION OF VEGETABLE BEANS AFTER VARIOUS PREDECESSORS USING RHIZOHUMIN
| null | 2,023
|
cc-by
| 5,969
|
УДК 633.1:631.5
DOI https://doi.org/10.32782/2310-0478-2023-1-26-32 Н. П. Cадовська
кандидат біологічних наук, доцент,
доцент кафедри плодоовочівництва і виноградарства
Ужгородський національний університет
(м. Ужгород, Україна)
E-mail: nadija.sadovska@uzhnu.edu.ua Г. Б. Попович
кандидат біологічних наук,
доцент кафедри плодоовочівництва і виноградарства
Ужгородський національний університет
(м. Ужгород, Україна)
E-mail: halina.popovich@uzhnu.edu.ua А. Ф. Гамор
кандидат біологічних наук, доцент,
доцент кафедри плодоовочівництва і виноградарства
Ужгородський національний університет
(м. Ужгород, Україна)
E-mail: andriy.hamor@uzhnu.edu.ua М. І. Опаленик
студент ІІ курсу магістратури
кафедри плодоовочівництва і виноградарства
Ужгородський національний університет
(м. Ужгород, Україна)
E-mail: opalenyk.mykhailo@student.uzhnu.edu.ua ФОРМУВАННЯ УРОЖАЮ КВАСОЛІ ОВОЧЕВОЇ
ПІСЛЯ РІЗНИХ ПОПЕРЕДНИКІВ
ЗА ВИКОРИСТАННЯ РИЗОГУМІНУ Наведено результати вивчення процесів росту та формування продуктивності квасолі овочевої при вирощуванні її після
культур-попередників картоплі та квасолі зернової за використання для посіву необробленого та обробленого біопрепа-
ратом «Ризогумін» насіння. Показано, що використання Ризогуміну для передпосівної обробки насіння не мало істотного
впливу на тривалість вегетації квасолі овочевої. У той же час найменша енергія проростання насіння (90,7%) була від-
мічена в контрольному варіанті на ділянці з культурою-попередником картоплею. На інших варіантах вона коливалася
від 94,7 до 97,3%. Очевидно, що приріст енергії проростання (від 4,0 до 7,3%) отримано за рахунок присутності абори-
генних штамів бульбочкових бактерій та наявності в дослідних варіантах бактеріальних клітин нових штамів. Найвищі
показники приросту вегетативної надземної маси отримано на варіанті, де насіння було оброблено біопрепаратом та
вирощувалося після попередника картоплі. Як кількість і розміри листків, так і площа асиміляційного апарату рослин
значно змінювалася за різних схем використання препарату. Кращі результати отримано за обробки насіння на ділянці,
де попередником слугувала картопля. Площа листкової поверхні була тут на 4,9% більшою, ніж за використання в якості
попередника бобової культури. Передпосівна інокуляція біопрепаратом спричиняла збільшення як загальної маси рос-
лин, так і її надземної частини та коренів. Разом із тим вищі показники отримано на ділянках, де квасолю вирощували
після картоплі. Найбільша кількість бобів (11 шт./росл.) формувалася на варіанті з висівом інокульованого насіння на
ділянці після картоплі. Відповідно на контролі їх утворювалося на 27,3% менше. На ділянці, де попередником була
квасоля зернова, у досліді на рослинах формувалося на 10% більше плодів, ніж у контролі. Використання Ризогуміну
призводило до зростання врожайності як за вирощування після картоплі, так і після квасолі зернової. Але показники
загальної (8,1 т/га) і товарної (7,9 т/га) урожайності були вищі на варіанті, де попередником слугувала бобова культура. Після попередника картоплі – відповідно 7,9 т/га та 7,7 т/га. Разом із тим приріст товарної врожайності до контролю був
вищим після картоплі 1,8 т/га проти 1,5 т/га. щ
р
,
/
р
,
/
Ключові слова: квасоля, насіння, Ризогумін, попередники, урожайність. AGRONOMY AGRONOMY УДК 633.1:631.5
DOI https://doi.org/10.32782/2310-0478-2023-1-26-32 CROP FORMATION OF VEGETABLE BEANS AFTER VARIOUS PREDECESSORS USING
RHIZOHUMIN The results of the study of the growth processes and the formation of the productivity of vegetable beans growing after the
predecessor crops of potatoes and grain beans using untreated and treated sowing seeds by biological preparation Rhizohumin. It was shown that using of Rhizohumin for pre-sowing seed treatment didn`t have a significant effect of the growing season
duration of vegetable beans. At the same time, the lowest energy of seed germination (90.7%) was noted in the control
variant on the area with the potato precursor culture. In other variants, it ranged from 94.7 to 97.3%. It is obvious that the
increase of germination energy (from 4.0 to 7.3%) was obtained due to the presence of aboriginal strains of nodule bacteria
and the presence of new strains of bubble bacteria in experimental versions of bacterial cells. In the variant where the seeds
were treated by biopreparation and grown after the predecessor of the potatoes the highest rates of increase in vegetative
above-ground mass were obtained. The best results were obtained with processing seeds in the area where potatoes served as
a predecessor. The area of the leaf surface was 4.9% greater than when using a legume precursor. Pre-sowing inoculation by
the biological preparation caused an increase in both the total mass of the plants, as well as its above-ground part and roots. At the same time, higher indicators were obtained in areas where beans were grown after potatoes. The largest number of
beans (11 pcs./plant) was formed on the variant with sowing of inoculated seeds on the area after potatoes. Accordingly, in the
control its were formed by 27.3% less. On the site where the precursor was grain beans in the experiment, 10% more fruits
were formed on the plants than in the control. Using of Rhizohumin led to an increase the productivity both when growing after
potatoes and after grain beans. But the indicators of total (8.1 t/ha) and marketable (7.9 t/ha) yield were higher in the variant
where leguminous crops served as a precursor. After the predecessor of potatoes – 7.9 t/ha and 7.7 t/ha, respectively. At the
same time, the increase in marketable productivity compared the control was higher after potatoes, 1.8 t/ha versus 1.5 t/ha. Key words: beans, seeds, Rhizohumin, predecessors, productivity. Постановка проблеми. Значне зменшення
виробництва високобілкових продуктів тварин-
ництва в нашій країні ставить завдання забез-
печення збалансованого харчування населення
білковими продуктами рослинного походження. N. P. Sadovska PhD of Biological Sciences, Associate Professor,
Associate Professor at the Department of Fruit and Vegetable Cultivation and Viticulture
Uzhhorod National University (Uzhhorod, Ukraine)
E-mail: nadija.sadovska@uzhnu.edu.ua BULLETIN OF UMAN NATIONAL UNIVERSITY OF HORTICULTURE № 1, 2023 № 1, 2023 26 АГРОНОМІЯ A. F. Hamor M. I. Opalenyk p
y
2nd year Master’s Student at the Department of Fruit and Vegetable Cultivation and Viticulture
Uzhhorod National University (Uzhhorod, Ukraine)
E-mail: opalenyk.mykhailo@student.uzhnu.edu.ua H. B. Popovych H. B. Popovych
PhD of Biological Sciences,
Associate Professor at the Department of Fruit and Vegetable Cultivation and Viticulture
Uzhhorod National University (Uzhhorod, Ukraine)
E-mail: halina.popovich@uzhnu.edu.ua opo yc
PhD of Biological Sciences,
Associate Professor at the Department of Fruit and Vegetable Cultivation and Viticulture
Uzhhorod National University (Uzhhorod, Ukraine)
E-mail: halina.popovich@uzhnu.edu.ua A. F. Hamor CROP FORMATION OF VEGETABLE BEANS AFTER VARIOUS PREDECESSORS USING
RHIZOHUMIN Добре росте
тільки на родючих, багатих на органічну речо-
вину, легких і чистих від бур’янів ґрунтах зі слаб-
кокислою або нейтральною реакцією ґрунтового
розчину. Разом із тим дослідження симбіотичної
діяльності рослин квасолі на дерново-підзолис-
тих важкосуглинкових ґрунтах показало, що
передпосівна інокуляція насіння «Ризобофітом»,
який містить симбіотичні азотфіксувальні бакте-
рії Rhizobium phaseoli, сприяла появі бульбочок,
збільшенню їх кількості, маси та активності нітро-
геназної системи [10]. У той же час внесення під-
вищених норм мінеральних добрив, зокрема до
N120P80K40, пригнічувало діяльність бульбочкових
бактерій. Докази негативного впливу завищених
норм мінеральних добрив на функціонування
бобово-ризобіальних систем наведено і в робо-
тах інших вчених [1; 2; 14]. У досліді закладали такі варіанти: Д1 – обробка
насіння квасолі перед висівом Ризогуміном, куль-
тура-попередник на ділянці – картопля; К1 – замо-
чування насіння перед висівом у воді, попередник
на ділянці – картопля; Д2 – обробка насіння квасолі
перед висівом Ризогуміном, попередник на ділянці –
квасоля зернова; К2 – замочування насіння перед
висівом у воді, попередник на ділянці – квасоля
зернова. Кожен варіант закладали в триразовому
повторенні. Площа облікової ділянки складала 6 м2. Дослідженнями [9] встановлено, що ефек-
тивність вирощування квасолі звичайної зрос-
тає за локального внесення навесні в зону май-
бутніх рядків (на 2–5 см нижче від розміщення
насіння) фосфорно-калійних добрив P30K30 та
інокулювання насіння «Ризобофітом» на тор-
фовій основі з кількістю бульбочкових бактерій
3,0–3,5 × 109 клітин в 1 г. Перед висівом насіння, призначене для
контрольних варіантів, зволожували у водогін-
ній воді з розрахунку 0,8–1,0% від маси насіння
та висівали. Насіння, призначене для дослід-
них варіантів, обробляли суспензією препарату
«Ризогумін». Для обробки насіння препарат
суспендували у водогінній воді, яка не містила
хлору. Оптимальна кількість води, згідно з реко-
мендаціями виробника [8], не повинна переви-
щувати 0,8–1,0% від маси насіння. Норма Ризо-
гуміну така: 200 г на 100 кг насіння. Висів на всіх
варіантах проводили у відкритий ґрунт за схемою
45×45 см на глибину 5 см по 5 насінин у лунку
в один день – 08.06.2021 р. На одній ділянці роз-
міщали по 15 кущів. ,
,
В останні роки роботи ряду вчених присвя-
чені вивченню впливу композицій біоінокулянтів
та мікоризоутворювальних препаратів на фор-
мування бобово-ризобіальних систем. Зокрема,
вченому В.В. Яценко [13] вдалося підібрати
ефективні симбіотичні поєднання для кращого
функціонування нодуляційного апарату рослин. Для квасолі спаржевої та бобів овочевих кращим
було поєднання препаратів «Андеріз» 2 л/т із
«Мікофрендом» 1,5 л/т та окреме застосування
інокулянту «Андеріз». CROP FORMATION OF VEGETABLE BEANS AFTER VARIOUS PREDECESSORS USING
RHIZOHUMIN У роботі [14] доведено позитивний вплив суміс-
ної або окремої інокуляції квасолі звичайної
Rhizobium tropici і Trichoderma harzianum, який
проявлявся у збільшенні кількості бульбочок та
підвищенні продуктивності рослин. У роботі [14] доведено позитивний вплив суміс-
ної або окремої інокуляції квасолі звичайної
Rhizobium tropici і Trichoderma harzianum, який
проявлявся у збільшенні кількості бульбочок та
підвищенні продуктивності рослин. значно нижча від біологічної. За умов потенцій-
ної врожайності сортів квасолі більш ніж 2,0 т/га
в окремі роки врожайність сягала всього 1,0 т/га,
а в середньому не перевищувала 1,7 т/га. Пра-
вильно підібраний сортимент дозволяє не лише
підвищити врожайність, але й покращити його
якість. Особливе місце сорту відводиться в енер-
гозберігаючих технологіях [6]. значно нижча від біологічної. За умов потенцій-
ної врожайності сортів квасолі більш ніж 2,0 т/га
в окремі роки врожайність сягала всього 1,0 т/га,
а в середньому не перевищувала 1,7 т/га. Пра-
вильно підібраний сортимент дозволяє не лише
підвищити врожайність, але й покращити його
якість. Особливе місце сорту відводиться в енер-
гозберігаючих технологіях [6]. Метою роботи є вивчення й порівняння
процесів росту та формування продуктивності
квасолі овочевої після різних попередників за
передпосівної обробки насіння біопрепаратом
«Ризогумін». Метою роботи є вивчення й порівняння
процесів росту та формування продуктивності
квасолі овочевої після різних попередників за
передпосівної обробки насіння біопрепаратом
«Ризогумін». Метою роботи є вивчення й порівняння
процесів росту та формування продуктивності
квасолі овочевої після різних попередників за
передпосівної обробки насіння біопрепаратом
«Ризогумін». р
[ ]
У роботі [11] наведено результати дослі-
джень впливу сортових особливостей квасолі на
утворення та функціонування бобово-ризобіаль-
них систем упродовж вегетації рослин. Показано,
що між азотфіксувальною здатністю та продук-
тивністю симбіотичних систем квасолі існує пряма
залежність, у результаті чого комплементарна дія
рослини-господаря та мікросимбіонта підвишує
активність процесів обміну речовин і зумовлює
підвищення продуктивності зазначених систем. Методика дослідження. Дослідження прово-
дилися у 2021 р. у приватному господарстві в ґрун-
тово-кліматичних умовах передгірної зони Закар-
паття. Вивчали вплив біопрепарату «Ризогумін»
на ростові процеси та формування врожаю квасолі
овочевої сорту Контендер після різних попередни-
ків за передпосівної обробки насіння. Ризогумін –
біодобриво, яке застосовується для бактеризації
насіння квасолі з метою поліпшення азотного жив-
лення рослин і підвищення продуктивності куль-
тури. Комплексний вплив препарату на продукцій-
ний процес квасолі дозволяє рекомендувати його
застосування не тільки на нових місцях її вирощу-
вання. Достовірний ефект інокуляції, як зазначає
виробник [8], забезпечується і при вирощуванні
культури на ґрунтах з високою щільністю популя-
цій аборигенних бульбочкових бактерій. Квасоля вимоглива до ґрунтів. CROP FORMATION OF VEGETABLE BEANS AFTER VARIOUS PREDECESSORS USING
RHIZOHUMIN Через це особлива увага повинна приділятися
проблемі збільшення валових зборів білка зер-
нобобових культур, особливо квасолі (Phaseolus
vulgaris L.) [4]. Вирощування квасолі зумовлене
як економічною, так і агрономічною привабли-
вістю. Квасоля, як і решта бобових культур,
збільшує вміст азоту в ґрунті та збагачує його
макро- і мікроелементами, що робить її надзви-
чайно корисним компонентом сівозміни, а також
одним із найкращих попередників для зернової
групи [10]. Для України квасоля є традиційною
культурою. Проте за останні 40–50 років її посівні
площі значно скоротилися. Для досягнення сві-
тового рівня виробництва квасолі потрібні висо-
копродуктивні та адаптовані до екстремальних
умов сорти. Наразі в Державний реєстр сортів,
придатних для поширення в Україні, занесено
понад 22 високопродуктивні сорти як вітчизня-
ної, так і зарубіжної селекції [6]. досить незначне бульбочкоутворення за раху-
нок спонтанного аборигенного інокулювання. Азотфіксувальний потенціал симбіозу квасолі
з присутніми в ґрунті ризобіями часто обмежений
невисокою азотфіксувальною активністю бакте-
рій [11]. У зв’язку з цим обов’язковим заходом
у технології вирощування квасолі повинна бути
передпосівна обробка насіння біопрепаратами
на основі селекціонованих штамів специфічних
ризобій, яка підвищує продуктивність рослин. Інокуляцію насіння активними штамами буль-
бочкових азотфіксувальних бактерій доцільно
проводити ще й через вартісність мінеральних
добрив, що збільшує витрати на вирощування
квасолі та її собівартість [3]. Аналіз останніх досліджень і публіка-
цій. За площами посіву квасоля наразі займає
2-ге місце у світі після сої. Основну концентра-
цію посівів зосереджено в Азії та Південній Аме-
риці. Незважаючи на те, що для України квасоля
вже давно є традиційною культурою, площі під
її посівами значно скоротилися. Згідно з даними
служби Державної статистики України «Посівні
площі сільськогосподарських культур під урожай
у 2019 р.» [7], площа під посівами квасолі стано-
вила 42,0 тис. га, із них 32,0 тис. га – у господар-
ствах населення і 9,6 тис. га – у сільгосппідприєм-
ствах. За останні роки фактична продуктивність
квасолі в усіх категоріях господарств України Попри перспективи розширення асортименту
сортів квасолі, азотфіксувальний потенціал цієї
культури повною мірою не використовується,
оскільки для квасолі звичайної в більшій мірі,
ніж для інших зернобобових культур, характерне ВІСНИК УМАНСЬКОГО НАЦІОНАЛЬНОГО УНІВЕРСИТЕТУ САДІВНИЦТВА ВІСНИК УМАНСЬКОГО НАЦІОНАЛЬНОГО УНІВЕРСИТЕТУ САДІВНИЦТВА № 1, 2023 № 1, 2023 27 AGRONOMY У роботі [14] доведено позитивний вплив суміс-
ної або окремої інокуляції квасолі звичайної
Rhizobium tropici і Trichoderma harzianum, який
проявлявся у збільшенні кількості бульбочок та
підвищенні продуктивності рослин. CROP FORMATION OF VEGETABLE BEANS AFTER VARIOUS PREDECESSORS USING
RHIZOHUMIN Доведено, що перебіг про-
дукційних процесів істотно залежить від сортових
особливостей та мікробіологічних препаратів. Дослідженнями [15] встановлено, що ризобакте-
рії, які сприяють росту ендофітних рослин (PGPR),
а саме штами Paenibacillus polymyxa та Bacillus
megaterium за спільної інокуляції з Rhizobium
проявляли синергетичний ефект на ріст бобів. Як вважають автори статті, використання PGPR
може підвищити ефективність біодобрив Rizobia. Впродовж вегетації проводили фенологічні
спостереження та біометричні виміри рослин. У міру формування бобів проводили їх збір для
визначення врожайності. Закладання дослідів
проводили згідно із загальноприйнятими методи-
ками [5]. Статистичну обробку результатів дослі-
джень здійснювали за допомогою комп’ютерної
програми Microsoft Office Excel 2016. Основні
результати
дослідження. У досліді перші сходи з’явилися на всіх варіантах
через 7 діб після висіву. Відчутної різниці в появі
повних сходів теж не відмічали. У той же час було BULLETIN OF UMAN NATIONAL UNIVERSITY OF HORTICULTURE BULLETIN OF UMAN NATIONAL UNIVERSITY OF HORTICULTURE № 1, 2023 28 АГРОНОМІЯ Для інтенсивного проходження фотосин-
тезу та формування високого врожаю бобів рос-
лини квасолі мають бути забезпечені оптималь-
ною площею листкової поверхні. Встановлено,
що найбільша кількість листків формувалася на
варіантах із використанням Ризогуміну (варіанти
Д1 і Д2), де насіння перед висівом було інокульо-
ване (табл. 2). Різниця в кількості сформованих
листків на ділянках, де попередником була кар-
топля, сягала 3 шт./росл. на користь варіанта
з насінням, попередньо обробленим Ризогуміном. На ділянках, де попередником була квасоля зер-
нова, ця різниця не перевищувала 0,7 шт./росл. встановлено, що найменша енергія проростання
насіння на варіанті із замочуванням насіння перед
висівом у воді (попередник картопля) сягала
90,7%, а на інших варіантах коливалась від 94,7
до 97,3%. Можна припустити, що приріст енергії
проростання (від 4,0 до 7,3%) отримано за раху-
нок присутності аборигенних штамів бульбочко-
вих бактерій та наявності в дослідних варіантах,
де використовувався Ризогумін, бактеріальних
клітин нових штамів. Тривалість вегетаційного
періоду коливалася від 93 діб на дослідних варі-
антах до 94–96 діб – на контрольних. Висота рослин у фазі бутонізації знаходи-
лася в межах 30,2–36,0 см (табл. 1). Найбільша
висота куща спостерігалася на варіанті Д1. У фазі
цвітіння цей показник збільшився на 1,4–2,0 см
порівняно з попереднім заміром. У період плодо-
ношення висота кущів зросла на 6,8–1,6 см. Най-
вищі кущі в кінці проведення вимірювань були на
ділянці, де насіння квасолі перед висівом було
оброблене Ризогуміном (варіант Д1), а найнижчі –
на ділянці, де насіння перед висівом замочува-
лося у воді (варіант К2). Діаметр кущів у фазі
бутонізації знаходився в межах 35,6–39,6 см. CROP FORMATION OF VEGETABLE BEANS AFTER VARIOUS PREDECESSORS USING
RHIZOHUMIN У фазі цвітіння він зріс на 2,4–3,4 см, а у фазі
плодоношення – ще на 6,6–9,8 см. В останній
фазі діаметр куща на варіанті Д1 на 15,9% пере-
вищував мінімальні розміри цього показника,
зафіксовані у випадку, коли насіння замочува-
лося перед висівом у воді (варіант К1). Максимальні
величини
площі
листкової
поверхні були зафіксовані на варіантах Д1,
Д2. Різниця між Д1 і К1 на ділянці, де культу-
рою-попередником слугувала картопля, сягала
11,7%. Відповідно різниця між Д2 і К2 (де попе-
редньо вирощували квасолю зернову) сягала
18,6%. Перерахунок площі листкової поверхні
рослин на гектар дав змогу оцінити площу асимі-
ляційного апарату агроценозу квасолі овочевої. Максимальним цей показник був на ділянках,
де посів проводили попередньо інокульованим
Ризогуміном насінням. Причому на ділянці, де
попередником була картопля, площа фотосин-
тезуючої поверхні була на 4,9% більшою. Різ-
ниця за величиною цього параметра між варіан-
тами Д1 і К1 сягала 1573,49 м2, а різниця між Д2
і К2 – 2400,37 м2. Таблиця 1
Біометричні параметри рослин квасолі овочевої у генеративному періоді
за різних схем вирощування
Варіант
Висота, см
Діаметр куща, см
бутонізація
цвітіння
плодоношення
бутонізація
цвітіння
плодоношення
Д1
36,0
38,0
44,8
39,6
43,0
52,8
К1
30,2
32,6
43,2
35,4
37,8
44,4
Д2
32,4
33,8
43,8
37,2
40,8
48,4
К2
32,0
33,4
42,2
35,6
38,0
45,2
НІР05
1,1
0,9
1,4
1,8
0,6
0,8 Біометричні параметри рослин квасолі овочевої у генеративному період
за різних схем вирощування Примітки: Д1 – обробка насіння квасолі перед висівом Ризогуміном, попередник на ділянці – картопля; К1 – замо-
чування насіння перед висівом у воді, попередник на ділянці – картопля; Д2 – обробка насіння квасолі перед висівом
Ризогуміном, попередник на ділянці – квасоля зернова; К2 – замочування насіння перед висівом у воді, попередник на
ділянці – квасоля зернова Примітки: Д1 – обробка насіння квасолі перед висівом Ризогуміном, попередник на ділянці – картопля; К1 – замо-
чування насіння перед висівом у воді, попередник на ділянці – картопля; Д2 – обробка насіння квасолі перед висівом
Ризогуміном, попередник на ділянці – квасоля зернова; К2 – замочування насіння перед висівом у воді, попередник на
ділянці – квасоля зернова Примітки: Д1 – обробка насіння квасолі перед висівом Ризогуміном, попередник на ділянці – картопля; К1 – замо-
чування насіння перед висівом у воді, попередник на ділянці – картопля; Д2 – обробка насіння квасолі перед висівом
Ризогуміном, попередник на ділянці – квасоля зернова; К2 – замочування насіння перед висівом у воді, попередник на
ділянці – квасоля зернова Примітки: Д1 – обробка насіння квасолі перед висівом Ризогуміном, попередник на ділянці – картопля; К1 – замо-
чування насіння перед висівом у воді, попередник на ділянці – картопля; Д2 – обробка насіння квасолі перед висівом
Ризогуміном, попередник на ділянці – квасоля зернова; К2 – замочування насіння перед висівом у воді, попередник на
ділянці – квасоля зернова ВІСНИК УМАНСЬКОГО НАЦІОНАЛЬНОГО УНІВЕРСИТЕТУ САДІВНИЦТВА ВІСНИК УМАНСЬКОГО НАЦІОНАЛЬНОГО УНІВЕРСИТЕТУ САДІВНИЦТВА
№ 1, 2023 Таблиця 2 Таблиця 2
Біометричні параметри асиміляційного апарату квасолі овочевої (фаза цвітіння)
Варіант
Кількість листків,
шт./росл. Розміри листка,
см
Площа листків,
см2/росл
Площа листкової поверхні,
тис. м2/га
Д1
12,0
10,0 × 6,16
547,0
13,51
К1
9,0
10,75 × 6,75
483,26
11,93
Д2
11,0
10,25 × 6,25
521,47
12,87
К2
10,3
9,25 × 6,0
424,25
10,47
НІР05
0,9
-
22,14
0,59
Примітки: Д1 – обробка насіння квасолі перед висівом Ризогуміном попередник на ділянці – картопля; К1 – замо- Табли
ометричні параметри асиміляційного апарату квасолі овочевої (фаза цвітіння) № 1, 2023 29 AGRONOMY AGRONOMY Найбільша кількість бобів формувалася на
варіанті Д1 – 11 шт. (табл. 3). На контролі К1 їх
було на 27,3% менше. На ділянці, де попередни-
ком була квасоля зернова, формувалося на 10%
більше плодів, ніж у контролі Використання Ризогуміну більшою мірою
впливало на висоту надземної частини рослин,
ніж на довжину кореневої системи. Найбільше
переважала надземна маса над масою кореневої
системи на варіанті Д1, де їх співвідношення було
5,5 : 1. . Довжина плоду з усіх варіантів суттєво не від-
різнялась і була в межах 14,5–15,5 см, а ширина
й маса плодів були однаковими. Урожай квасолі овочевої збирали багатора-
зово. Усього провели 8 зборів (табл. 5). Особливо продуктивними були 2-ий – 5-ий
збори. Приріст урожаю у 2-му зборі коливався від
96,5% до 139%. Найбільшим він був на варіанті
Д1. Мінімальний приріст відмічено на варіанті Д2. Максимальну масу плодів було отримано з 3-го Найбільшу масу мали рослини на тих ділян-
ках, де застосовувався препарат «Ризогумін». Подібне співвідношення спостерігали і при зва-
жуванні надземної та кореневої частин квасолі
(табл. 4). Таблиця 3
Біометричні параметри плодів квасолі овочевої у технічній стиглості
Варіант
Кількість плодів, шт./росл. Довжина плоду, см
Ширина плоду, шт. Примітки: Д1 – обробка насіння квасолі перед висівом Ризогуміном, попередник на ділянці – картопля; К1 – замо-
чування насіння перед висівом у воді, попередник на ділянці – картопля; Д2 – обробка насіння квасолі перед висівом
Ризогуміном, попередник на ділянці – квасоля зернова; К2 – замочування насіння перед висівом у воді, попередник на
ділянці – квасоля зернова BULLETIN OF UMAN NATIONAL UNIVERSITY OF HORTICULTURE Таблиця 2 Маса, г
Д1
11,0
15,5
0,8
7,0
К1
8,0
15,0
0,8
7,0
Д2
10,0
15,0
0,8
7,0
К2
9,0
14,5
0,8
7,0
НІР05
0,9
0,02
-
-
Примітки: Д1 – обробка насіння квасолі перед висівом Ризогуміном, попередник на ділянці – картопля; К1 – замо-
чування насіння перед висівом у воді, попередник на ділянці – картопля; Д2 – обробка насіння квасолі перед висівом
Ризогуміном, попередник на ділянці – квасоля зернова; К2 – замочування насіння перед висівом у воді, попередник на
ділянці – квасоля зернова Біометричні параметри плодів квасолі овочевої у технічній стиглості Примітки: Д1 – обробка насіння квасолі перед висівом Ризогуміном, попередник на ділянці – картопля; К1 – замо-
чування насіння перед висівом у воді, попередник на ділянці – картопля; Д2 – обробка насіння квасолі перед висівом
Ризогуміном, попередник на ділянці – квасоля зернова; К2 – замочування насіння перед висівом у воді, попередник на
ділянці – квасоля зернова BULLETIN OF UMAN NATIONAL UNIVERSITY OF HORTICULTURE № 1, 2023
BULLETIN OF UMAN NATIONAL UNIVERSITY OF HORTICULTURE Література Продовольча індустрія АПК. 2015. № 1–2. С. 38–40. http://nbuv.gov.ua/UJRN/
Piapk_2015_1-2_10 5. Основи наукових досліджень в агрономії : під-
ручник / В.О. Єщенко, П.Г. Копитко, П.В. Костогриз,
В.П. Опришко / за ред. В.О. Єщенка. Вінниця : ПП
«ТД "Едельвейс і К"», 2014. 332 с. Висновки. Біопрепарат
«Ризогумін»
за
передпосівної обробки насіння істотно не впли-
вав на тривалість вегетації квасолі овочевої. Разом із тим значно змінювалися біометричні
параметри рослин (висота, діаметр, кількість та
розміри листків, площа асиміляційної поверхні)
залежно від варіанта. Кращі показники отри-
мано на варіанті, де насіння було оброблено біо-
препаратом «Ризогумін» та вирощувалося після
попередника картоплі. Зокрема, площа фотосин-
тезувальної поверхні була тут на 4,9% більшою,
ніж за використання в якості попередника бобо-
вої культури. На цьому ж варіанті більшими були
розміри та маса надземної частини й кореневої
системи рослин. 6. Особливості прояву господарсько-біологіч-
них ознак квасолі звичайної (Phaseolus vulgaris L.)
в умовах Лісостепу Правобережного : монографія /
В.А. Мазур та ін. Вінниця : ТОВ «Друк», 2021. 256 с. 6. Особливості прояву господарсько-біологіч-
них ознак квасолі звичайної (Phaseolus vulgaris L.)
в умовах Лісостепу Правобережного : монографія /
В.А. Мазур та ін. Вінниця : ТОВ «Друк», 2021. 256 с. 7. Посівні площі сільськогосподарських культур
під урожай у 2019 р. Державна служба cтатистики
України. https://ukrstat.gov.ua 7. Посівні площі сільськогосподарських культур
під урожай у 2019 р. Державна служба cтатистики
України. https://ukrstat.gov.ua 8. Ризогумін (торф’яний) для квасолі / Інститут
сільськогосподарської мікробіології та агропромис-
лового виробництва НААН. https://ismav.com.ua/
produkciya/biopreparati-dlya-roslinnictva/rizogumin-
torfyanij-kvasolya/ 8. Ризогумін (торф’яний) для квасолі / Інститут
сільськогосподарської мікробіології та агропромис-
лового виробництва НААН. https://ismav.com.ua/
produkciya/biopreparati-dlya-roslinnictva/rizogumin-
torfyanij-kvasolya/ Використання Ризогуміну призводило до
зростання врожайності як за вирощування після
картоплі, так і після квасолі зернової. Але вищі
показники загальної й товарної врожайності
отримано на варіанті, де попередником слугу-
вала бобова культура. Відповідно врожайність на
цьому варіанті сягала 8,1 т/га та 7,9 т/га, у той
час як після попередника картоплі – 7,9 т/га та
7,7 т/га. Разом із тим приріст товарної врожай-
ності до контролю був вищим після попередника
картоплі – 1,8 т/га проти 1,5 т/га. 9. Сайко О.Ю., Носенко Ю.М. Ефективний спо-
сіб вирощування квасолі звичайної. Овочівництво і
баштанництво. 2015. № 61. С. 200–206. 10. Цибрій-Сівак Н.В., Бахмат М.І. Формування
продуктивності квасолі звичайної залежно від сор-
тів, інокуляції та удобрення. Сільськогосподарські
науки. 2022. Вип. 2(37). С. 32–40. DOI: https://doi. org/10.37406/2706-9052-2022-2-5 10. Цибрій-Сівак Н.В., Бахмат М.І. Формування
продуктивності квасолі звичайної залежно від сор-
тів, інокуляції та удобрення. Сільськогосподарські
науки. 2022. Вип. 2(37). С. 32–40. DOI: https://doi. Таблиця 6 Загальна та товарна врожайність квасолі овочевої Таблиця 6
Загальна та товарна врожайність квасолі овочевої
Варіант
Урожайність, т/га
Приріст товарної врожайності до контролю
загальна
товарна
т/га
%
Д1
7,9
7,7
1,8
23,3
К1
6,2
5,9
-
-
Д2
8,1
7,9
1,5
23,4
К2
6,7
6,4
-
-
НІР05
0,20
0,16
-
- Приріст товарної врожайності до контрол Примітки: Д1 – обробка насіння квасолі перед висівом Ризогуміном, попередник на ділянці – картопля; К1 – замо-
чування насіння перед висівом у воді, попередник на ділянці – картопля; Д2 – обробка насіння квасолі перед висівом
Ризогуміном, попередник на ділянці – квасоля зернова; К2 – замочування насіння перед висівом у воді, попередник на
ділянці – квасоля зернова Примітки: Д1 – обробка насіння квасолі перед висівом Ризогуміном, попередник на ділянці – картопля; К1 – замо-
чування насіння перед висівом у воді, попередник на ділянці – картопля; Д2 – обробка насіння квасолі перед висівом
Ризогуміном, попередник на ділянці – квасоля зернова; К2 – замочування насіння перед висівом у воді, попередник на
ділянці – квасоля зернова Література Література збору (09.08). Найменшим був урожай із пере-
достаннього збору, проведеного в кінці серпня. 1. Доктор Н.М., Новицька Н.В., Кормош С.М.,
Пилипенко В.С., Мартинов О.В. Урожайність квасолі
звичайної (Phaseolus vulgaris L.) залежно від іно-
куляції та удобрення в умовах Закарпаття України. Аграрні інновації. 2022. № 13. DOI: https://doi. org/10.32848/agrar.innov.2022.13.8 Загальний урожай зелених плодів з обліко-
вих ділянок коливався від 3,73 до 4,84 кг. На
ділянці Д1 приріст до контролю (К1) склав 1,02 кг
(27,3%). На варіанті Д2 приріст до контролю (К2)
був менший і сягав 0,85 кг (21,3%). у
(
)
Загальна та товарна врожайність зелених
бобів квасолі після перерахунку на одиницю
площі наведена в таблиці 6. 2. Новицька Н.В., Мартинов О.М., Доктор Н.М. Вегетація квасолі під впливом передпосівної іно-
куляції насіння та удобрення. Вісник Полтавської
державної аграрної академії. 2018. № 2. С. 45–48. DOI: https://doi.org/10.31210/visnyk2018.02.07 2. Новицька Н.В., Мартинов О.М., Доктор Н.М. Вегетація квасолі під впливом передпосівної іно-
куляції насіння та удобрення. Вісник Полтавської
державної аграрної академії. 2018. № 2. С. 45–48. DOI: https://doi.org/10.31210/visnyk2018.02.07 Найвищу врожайність було отримано на варі-
анті Д2 (8,1 т/га). На ділянці Д1 загальна врожай-
ність була нижчою на 0,2 т. Приріст загальної
врожайності на варіанті Д1 по відношенню до К1
склав 1,7 т/га. Зростання врожайності на варіанті
Д2 по відношенню до К2 – 1,4 т/га. Товарна вро-
жайність на всіх варіантах досліду сягала понад
95% від загальної. Приріст товарної врожайності
відносно контролю К1 на ділянці Д1 склав 1,8 т/га
(23,3%). Величина приросту на ділянці Д2 сягала
1,3 т/га (23,4%). Найвищу врожайність було отримано на варі-
анті Д2 (8,1 т/га). На ділянці Д1 загальна врожай-
ність була нижчою на 0,2 т. Приріст загальної
врожайності на варіанті Д1 по відношенню до К1
склав 1,7 т/га. Зростання врожайності на варіанті
Д2 по відношенню до К2 – 1,4 т/га. Товарна вро-
жайність на всіх варіантах досліду сягала понад
95% від загальної. Приріст товарної врожайності
відносно контролю К1 на ділянці Д1 склав 1,8 т/га
(23,3%). Величина приросту на ділянці Д2 сягала
1,3 т/га (23,4%). 3. Носенко Ю. Товарне вирощування квасолі зви-
чайної. Агрономія сьогодні. 2015. № 9(304). http://
agro-business.com.ua/ahrarni-kultury/item/554-
tovarne-vyroshchuvannia-kvasoli-zvychainoi.html 3. Носенко Ю. Товарне вирощування квасолі зви-
чайної. Агрономія сьогодні. 2015. № 9(304). http://
agro-business.com.ua/ahrarni-kultury/item/554-
tovarne-vyroshchuvannia-kvasoli-zvychainoi.html y
y
4. Овчарук О., Іванюк С. Квасоля – цінне
джерело
рослинного
білка,
зумовлене
сорто-
вими особливостями. Продовольча індустрія АПК. 2015. № 1–2. С. 38–40. http://nbuv.gov.ua/UJRN/
Piapk_2015_1-2_10 y
y
4. Овчарук О., Іванюк С. Квасоля – цінне
джерело
рослинного
білка,
зумовлене
сорто-
вими особливостями. ВІСНИК УМАНСЬКОГО НАЦІОНАЛЬНОГО УНІВЕРСИТЕТУ САДІВНИЦТВА Таблиця 4 Таблиця 4
Маса та розміри рослин квасолі овочевої
Варіант
Загальна
маса
рослини, г
Маса
надземної
частини,г
Маса
кореневої
системи, г
Висота
надземної
частини, см
Довжина
кореневої
системи, см
Співвід-
ношення маси
надземної
частини до
коренів
Д1
55,4
46,9
8,5
38,3
28,8
5,5 : 1
К1
47,6
39,6
8,0
33,8
27,0
5,0 : 1
Д2
52,3
44,0
8,3
36,0
28,5
5,3 : 1
К2
45,2
38,1
7,1
33,4
27,4
5,4 : 1
НІР05
2,5
3,2
0,4
2,8
0,2
-
Примітки: Д1 – обробка насіння квасолі перед висівом Ризогуміном, попередник на ділянці – картопля; К1 – замо-
чування насіння перед висівом у воді, попередник на ділянці – картопля; Д2 – обробка насіння квасолі перед висівом
Ризогуміном, попередник на ділянці – квасоля зернова; К2 – замочування насіння перед висівом у воді, попередник на
ділянці – квасоля зернова Примітки: Д1 – обробка насіння квасолі перед висівом Ризогуміном, попередник на ділянці – картопля; К1 – замо-
чування насіння перед висівом у воді, попередник на ділянці – картопля; Д2 – обробка насіння квасолі перед висівом
Ризогуміном, попередник на ділянці – квасоля зернова; К2 – замочування насіння перед висівом у воді, попередник на
ділянці – квасоля зернова Таблиця 5
Маса плодів квасолі (у технічній стиглості) з окремих зборів та загальний урожай
Варіант
Дата збору та маса зібраних плодів, г
Урожай
з ділянки, кг
31.07
05.08
09.08
13.08
18.08
23.08
27.08
02.09
Д1
385
920
1020
845
820
380
150
230
4,75
К1
305
645
840
690
650
250
140
210
3,73
Д2
425
835
1465
770
745
265
150
185
4,84
К2
345
740
885
660
710
290
155
200
3,99
НІР05
35
58
112
62
27
40
12
18
-
П
і
Д1
б
б
і
і
і
Р
і
і
і
К1 Таблиця
плодів квасолі (у технічній стиглості) з окремих зборів та загальний урожай Примітки: Д1 – обробка насіння квасолі перед висівом Ризогуміном, попередник на ділянці – картопля; К1 – замо-
чування насіння перед висівом у воді, попередник на ділянці – картопля; Д2 – обробка насіння квасолі перед висівом
Ризогуміном, попередник на ділянці – квасоля зернова; К2 – замочування насіння перед висівом у воді, попередник на
ділянці – квасоля зернова № 1, 2023 30 АГРОНОМІЯ Література org/10.37406/2706-9052-2022-2-5 g
11. Шкатула Ю.М., Краєвська Л.С. Ефективність
симбіотичної азотфіксації в агроценозах квасолі. ВІСНИК УМАНСЬКОГО НАЦІОНАЛЬНОГО УНІВЕРСИТЕТУ САДІВНИЦТВА ВІСНИК УМАНСЬКОГО НАЦІОНАЛЬНОГО УНІВЕРСИТЕТУ САДІВНИЦТВА № 1, 2023 № 1, 2023 31 AGRONOMY Вісник Дніпропетровського аграрно-економічного
університету. 2015. № 4(38). С. 73–76. Вісник Дніпропетровського аграрно-економічного
університету. 2015. № 4(38). С. 73–76. Вісник Дніпропетровського аграрно-економічного
університету. 2015. № 4(38). С. 73–76. 12. Шкатула Ю.М., Краєвська Л.С. Роль біоло-
гічного азоту в підвищенні насіннєвої продуктив-
ності квасолі. Сільське господарство та лісівництво. Вінниця, 2016. Вип. 4. С. 231–239. 13. Яценко
В.В. Сортові
особливості
фор-
мування нодуляційного апарату бобових куль-
тур за використання інокулянтів і мікоризоутво-
рювального препарату. Збірник наукових праць
Уманського національного університету садівни-
цтва. Умань, 2022. Вип. 100. Ч. 1. С. 100–114. DOI: 10.31395/2415-8240-2022-100-1-100-114 7. Posivni ploshchi silskohospodarskykh kultur
pid urozhai u 2019 r [Sown areas of agricultural crops
for harvest in 2019]. Derzhavna sluzhba ctatystyky
Ukrainy. Retrieved from URL: https://ukrstat.gov.ua
[in Ukrainian].i 7. Posivni ploshchi silskohospodarskykh kultur
pid urozhai u 2019 r [Sown areas of agricultural crops
for harvest in 2019]. Derzhavna sluzhba ctatystyky
Ukrainy. Retrieved from URL: https://ukrstat.gov.ua
[in Ukrainian].i 8. Ryzohumin (torfianyi) dlia kvasoli [Rhizohumin
(peat)
for
beans]. Instytut
silskohospodarskoi
mikrobiolohii ta ahropromyslovoho vyrobnytstva NAAN. Retrieved from URL: https://ismav.com.ua/produkciya/
biopreparati-dlya-roslinnictva/rizogumin-torfyanij-
kvasolya/ [in Ukrainian]. 14. Alice
M. Mweetwa,
Gwen
Chilombo
&
Brian M. Condwe. Nodulation, nutrient uptake and
yield of common bean inoculated with Rhizobia and
Trichoderma in an acid soil. Journal of Agriculnural
Science. 2016. Vol. 8. No 12. P. 61–71. DOI:10.5539/
jas.v8n12pxx 9. Saiko, O.Iu., Nosenko, Yu.M. (2015). Efektyvnyi
sposib vyroshchuvannia kvasoli zvychainoi [It’s an
effective way of cultivation of Phaseolus vulgaris
L.]. Ovochivnytstvo i bashtannytstvo, 61, 200–206
[in Ukrainian]. 9. Saiko, O.Iu., Nosenko, Yu.M. (2015). Efektyvnyi
sposib vyroshchuvannia kvasoli zvychainoi [It’s an
effective way of cultivation of Phaseolus vulgaris
L.]. Ovochivnytstvo i bashtannytstvo, 61, 200–206
[in Ukrainian]. 15. Korir H., Mungai N.W., Thuita M., Hamba Y. and Macco C. Co-inoculation Effect of Rhizobia and
Plant Growth Promoting Rhizobacteria on Common
Bean Growth in a Low Phosphorus Soil. Front. Plant
Sci. 2017. Vol. 8. Р. 141. https://doi.org/10.3389/
fpls.2017.00141 10. Tsybrii-Sivak, N.V., Bakhmat, M.I. (2022). Formuvannia
produktyvnosti
kvasoli
zvychainoi
zalezhno
vid
sortiv,
inokuliatsii
ta
udobrennia
[Forming the productivity of the common beans
dependingon varieties, inoculation, and fertilizer]. Silskohospodarski nauky, 2(37), 32–40. DOI: https://
doi.org/10.37406/2706-9052-2022-2-5 [in Ukrainian]. 6. Mazur, V.A., Didur, I.M., Mazur, O.V., Mazur, O.V.
(2021). Osoblyvosti proiavu hospodarsko-biolohichnykh
oznak kvasoli zvychainoi (Phaseolus Vulgaris L.) v
umovakh Lisostepu Pravoberezhnoho [Peculiarities
of the manifestation of economic and biological
characteristics of common beans (Phaseolus Vulgaris L.)
in the conditions of in Right-bank Ukraine Forest-
Steppe]. Vinnytsia: TOV «Druk», 256 s. [in Ukrainian]. References 1. Doktor, N.M., Novytska, N.V., Kormosh, S.M.,
Pylypenko, V.S., Martynov, O.V. (2022). Urozhainist
kvasoli zvychainoi (Phaseolus vulgaris L.) zalezhno
vid inokuliatsii ta udobrennia v umovakh Zakarpattia
Ukrainy [Yield of common beans (Phaseolus vulgaris L.)
depending on inoculation and fertilizer in the conditions
of Transcarpathia of Ukraine]. Ahrarni inovatsii,
13,
53–57. DOI:
https://doi.org/10.32848/agrar. innov.2022.13.8 [in Ukrainian]. 1. Doktor, N.M., Novytska, N.V., Kormosh, S.M.,
Pylypenko, V.S., Martynov, O.V. (2022). Urozhainist
kvasoli zvychainoi (Phaseolus vulgaris L.) zalezhno
vid inokuliatsii ta udobrennia v umovakh Zakarpattia
Ukrainy [Yield of common beans (Phaseolus vulgaris L.)
depending on inoculation and fertilizer in the conditions
of Transcarpathia of Ukraine]. Ahrarni inovatsii,
13,
53–57. DOI:
https://doi.org/10.32848/agrar. innov.2022.13.8 [in Ukrainian]. 11. Shkatula, Yu.M., Kraievska, L.S. (2015). Efektyvnist symbiotychnoi azotfiksatsii v ahrotsenozakh
kvasoli [Efficiency of symbiotic nitrogen fixation
in bean agrocenoses]. Visnyk Dnipropetrovskoho
ahrarno-ekonomichnoho universytetu, 4(38), 73–76
[in Ukrainian]. 11. Shkatula, Yu.M., Kraievska, L.S. (2015). Efektyvnist symbiotychnoi azotfiksatsii v ahrotsenozakh
kvasoli [Efficiency of symbiotic nitrogen fixation
in bean agrocenoses]. Visnyk Dnipropetrovskoho
ahrarno-ekonomichnoho universytetu, 4(38), 73–76
[in Ukrainian]. 2. Novytska, N.V., Martynov, O.M., Doktor, N.M. (2018). Vehetatsiia kvasoli pid vplyvom peredposivnoi
inokuliatsii nasinnia ta udobrennia [Haricot vegetation
under the influence of foreign intolution seeds and
approvals]. Visnyk Poltavskoi derzhavnoi ahrarnoi
akademii, 2, 45–48. DOI: https://doi.org/10.31210/
visnyk2018.02.07 [in Ukrainian]. 2. Novytska, N.V., Martynov, O.M., Doktor, N.M. (2018). Vehetatsiia kvasoli pid vplyvom peredposivnoi
inokuliatsii nasinnia ta udobrennia [Haricot vegetation
under the influence of foreign intolution seeds and
approvals]. Visnyk Poltavskoi derzhavnoi ahrarnoi
akademii, 2, 45–48. DOI: https://doi.org/10.31210/
visnyk2018.02.07 [in Ukrainian]. 12. Shkatula, Yu.M., Kraievska, L.S. (2016). Rol biolohichnoho azotu v pidvyshchenni nasinnievoi
produktyvnosti kvasoli [The role of biological nitrogen
in increasing seed productivity of beans]. Silske
hospodarstvo ta lisivnytstvo, 4, 231–239 [in Ukrainian]. 13. Yatsenko, V.V. (2022). Sortovi osoblyvosti
formuvannia noduliatsiinoho aparatu bobovykh kultur
za vykorystannia inokuliantiv i mikoryzoutvoriuvalnoho
preparatu [Variety features of formation of nodulation
apparatus of bean cultures using bioinoculants and micorise-
forming drug]. Zbirnyk naukovykh prats Umanskoho
natsionalnoho
universytetu
sadivnytstva,
100(1),
100–114. DOI:
10.31395/2415-8240-2022-100-1-
100-114 [in Ukrainian]. 3. Nosenko, Yu. (2015). Tovarne vyroshchuvannia
kvasoli zvychainoi [Commercial cultivation of common
beans]. Ahronomiia
sohodni,
9(304). Retrieved
from
URL
http://agro-business.com.ua/ahrarni-
kultury/item/554-tovarne-vyroshchuvannia-kvasoli-
zvychainoi.html [in Ukrainian]. 3. Nosenko, Yu. (2015). Tovarne vyroshchuvannia
kvasoli zvychainoi [Commercial cultivation of common
beans]. Ahronomiia
sohodni,
9(304). Retrieved
from
URL
http://agro-business.com.ua/ahrarni-
kultury/item/554-tovarne-vyroshchuvannia-kvasoli-
zvychainoi.html [in Ukrainian]. 4. Ovcharuk, O., Ivaniuk, S. (2015). References Kvasolia –
tsinne dzherelo roslynnoho bilka, zumovlene sortovymy
osoblyvostiamy [A kidney bean is the valuable source
of phytalbumin, conditioned of high quality features]. Prodovolcha industriia APK, 1–2, 38–40. Retrieved from
URL:
http://nbuv.gov.ua/UJRN/Piapk_2015_1-2_10
[in Ukrainian]. 4. Ovcharuk, O., Ivaniuk, S. (2015). Kvasolia –
tsinne dzherelo roslynnoho bilka, zumovlene sortovymy
osoblyvostiamy [A kidney bean is the valuable source
of phytalbumin, conditioned of high quality features]. Prodovolcha industriia APK, 1–2, 38–40. Retrieved from
URL:
http://nbuv.gov.ua/UJRN/Piapk_2015_1-2_10
[in Ukrainian]. [
]
14. Alice, M. Mweetwa, Gwen, Chilombo &
Brian, M. Condwe (2016). Nodulation, nutrient uptake
and yield of common bean inoculated with Rhizobia
and Trichoderma in an acid soil. Journal of Agriculnural
Science, 8(12), 61–71. DOI:10.5539/jas.v8n12pxx
15. Korir, H., Mungai, N.W., Thuita, M., Hamba, Y. and Masso, C. (2017). Co-inoculation Effect of Rhizobia
and Plant Growth Promoting Rhizobacteria on Common
Bean Growth in a Low Phosphorus Soil. Front. Plant Sci,
8, 141. https://doi.org/10.3389/fpls.2017.00141 14. Alice, M. Mweetwa, Gwen, Chilombo &
Brian, M. Condwe (2016). Nodulation, nutrient uptake
and yield of common bean inoculated with Rhizobia
and Trichoderma in an acid soil. Journal of Agriculnural
Science, 8(12), 61–71. DOI:10.5539/jas.v8n12pxx 5. Ieshchenko, V.O., Kopytko, P.H., Kostohryz, P.V.,
Opryshko, V.P. (2014). Osnovy naukovykh doslidzhen
v ahronomii [Fundamentals of scientific research in
agronomy]. Vinnytsia: PP "TD "Edelveis i K"". 332 s. [in Ukrainian]. 5. Ieshchenko, V.O., Kopytko, P.H., Kostohryz, P.V.,
Opryshko, V.P. (2014). Osnovy naukovykh doslidzhen
v ahronomii [Fundamentals of scientific research in
agronomy]. Vinnytsia: PP "TD "Edelveis i K"". 332 s. [in Ukrainian]. 15. Korir, H., Mungai, N.W., Thuita, M., Hamba, Y. and Masso, C. (2017). Co-inoculation Effect of Rhizobia
and Plant Growth Promoting Rhizobacteria on Common
Bean Growth in a Low Phosphorus Soil. Front. Plant Sci,
8, 141. https://doi.org/10.3389/fpls.2017.00141 BULLETIN OF UMAN NATIONAL UNIVERSITY OF HORTICULTURE № 1, 2023 № 1, 2023 32
|
https://openalex.org/W2954501945
|
https://nvlvet.com.ua/index.php/agriculture/article/download/3718/3692
|
Ukrainian
| null |
Prospects of transgenic plants in the agro-sphere of Lviv region
|
Naukovij vìsnik Lʹvìvsʹkogo nacìonalʹnogo unìversitetu veterinarnoï medicini ta bìotehnologìj ìmenì S.Z. Gžicʹkogo. Serìâ: Sìlʹsʹkogospodarsʹkì nauki/Naukovij vìsnik Lʹvìvsʹkogo nacìonalʹnogo unìversitetu veterinarnoï medicini ta bìotehnologìj ìmenì S.Z. G̀žicʹkogo. Serìâ Sìlʹsʹkogospodarsʹkì nauki
| 2,019
|
cc-by
| 3,978
|
Article info Received 06.02.2019
Received in revised form
11.03.2019
Accepted 12.03.2019 The state and prospects of the use of transgenic plants in agriculture of Lviv region are considered
in the paper. Transgenic plants are a kind of genetically modified organisms obtained by biotechnologi-
cal methods using trans-transfer (between unrelated species) genes. In recent years, such cultures have
become widespread in the world. The vast majority of soy, corn and other crops are planted with GM
varieties. In the EU countries there is no broad perception of such cultures and there are significant
restrictions on their distribution. Approximately 70 types of GMOs are allowed in the EU, including
many decorative products (flowers) and about 58 GMOs for use in food and feed. In the world, 168 lines
of genetically modified plants: corn, rape, cotton, potatoes, soya, rice, tomatoes, wheat and others were
registered and allowed to be used for industrial production of food and feed. Our country also has some
experience in obtaining transgenic organisms. According to unofficial data, in Ukraine transgenic 60–
70% soybeans, 10–20% corn, 5% rape. In the near future, Ukraine should develop a clear concept of
attitude towards GMOs, and the absence of a position on this issue complicates the planning of activities
of the agro-sector. In the Lviv region there are prospects for growing GM varieties of soybean, corn and
sunflower; there is evidence that a significant part of the crops of the first belongs to the modified varie-
ties. Growing of GM sunflower in comparison with the usual one will have limited competitive ad-
vantages. Of the other crops that the area is specialized in, one can name potatoes whose GM varieties
are used mainly for technical needs, and wheat, whose GM varieties have not yet been sufficiently tested. In the world market, there is a demand for genetically modified crops, an estimated 68% of Ukrainian
agroholdings claim to be willing and able to grow GM crops for their legalization in Ukraine. Stepan Gzhytskyi National
University of Veterinary Medicine
and Biotechnologies Lviv,
Pekarska Str., 50, Lviv,
79010, Ukraine. Tel.: +38-096-289-52-40
E-mail: kalynb@bigmir.net Key words: agrarian sector, transgenic plant organism, crop production, Lviv region. Key words: agrarian sector, transgenic plant organism, crop production, Lviv region. Stepan Gzhytskyi National
University of Veterinary Medicine
and Biotechnologies Lviv,
Pekarska Str., 50, Lviv,
79010, Ukraine.
Tel.: +38-096-289-52-40
E-mail: kalynb@bigmir.net Науковий вісник ЛНУВМБ імені С.З. Ґжицького. Серія: Сільськогосподарські науки, 2019, т 21, № 90 Науковий вісник ЛНУВМБ імені С.З. Ґжицького. Серія: Сільськогосподарські науки, 2019, т 21, № 90 Науковий вісник Львівського національного університету
ветеринарної медицини та біотехнологій імені С.З. Ґжицького. Серія: Сільськогосподарські науки
Scientific Messenger of Lviv National University
of Veterinary Medicine and Biotechnologies. Series: Agricultural sciences ISSN 2519–2698 print
https://nvlvet.com.ua/index.php/agriculture doi: 10.32718/nvlvet‐a9009 doi: 10.32718/nvlvet‐a9009 UDC 631.524 R.P. Paranjak, B.M. Kalyn, B.V. Gutyj nal University of Veterinary Medicine and Biotechnologies Lviv, Ukraine Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv, Ukraine Scientific Messenger LNUVMB. Series: Agricultural sciences, 2019, vol. 21, no 90 Key words: agrarian sector, transgenic plant organism, crop production, Lviv region Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv, Ukraine Paranjak, R.P., Kalyn, B.M., & Gutyj, B.V. (2019). Prospects of transgenic plants in the
agro-sphere of Lviv region. Scientific Messenger of Lviv National University of Veterinary
Medicine and Biotechnologies. Series: Agricultural sciences, 21(90), 54–58. doi: 10.32718/nvlvet-
a9009 Paranjak, R.P., Kalyn, B.M., & Gutyj, B.V. (2019). Prospects of transgenic plants in the
agro-sphere of Lviv region. Scientific Messenger of Lviv National University of Veterinary
Medicine and Biotechnologies. Series: Agricultural sciences, 21(90), 54–58. doi: 10.32718/nvlvet-
a9009 ваних заходів для забезпечення належного рівня захи-
сту в галузі використання сучасних біотехнологій і,
зокрема, ГМО. ваних заходів для забезпечення належного рівня захи-
сту в галузі використання сучасних біотехнологій і,
зокрема, ГМО. Використання трансгенних організмів є одним із
найбільш контраверсійних питань сьогодення. Прак-
тика їхнього використання у розвинених країнах має
виражений господарсько-економічний ефект. Виро-
щування таких рослин дозволяє знизити обсяги засто-
сування пестицидів і збільшити врожайність. Разом із
тим з погляду значення для екологічних спільнот
використання у широкій практиці, поширення нових
організмів, які належать до трансгенних, є питанням,
котре не можна ігнорувати попри те, що більшість
проблем наразі носять потенційний характер. Перші трансгенні рослини були одержані у
1983 р.; перший харчовий ГМ продукт (сир) – вигото-
влений із використанням генетично модифікованого
ферменту – був дозволений у США в 1990 р. (Rudyshyn, 2011). Початок комерційним посівам тран-
сгенних сортів поклали не так давно – у 1996 р., але
вже сьогодні вони займають кожен десятий гектар у
світі. Незважаючи на опозицію у певних колах, нові
сорти трансгенних рослин швидко завойовують попу-
лярність у світі; зокрема, площа під чотирма найбільш
поширеними культурами (соя, кукурудза, бавовник,
ріпак) складає майже 30% світових посівів. Серед
країн, де найбільші площі зайняті трансгенними куль-
турами, називають передусім країни західної півкулі:
США, Канада, Аргентина, Бразилія. Чимало таких
культур у Китаї, нещодавно (з 2011 року) Індія знач-
ною мірою перейшла на використання генетично-
модифікованої бавовни. У багатьох державах генно-
інженерні сорти переважають над традиційними. Так,
у 2010 р. 93% площ сої в США, 99% в Аргентині, 75%
у Бразилії висіяли ГМ-насінням. Для кукурудзи це –
відповідно 86%, 86% та 56%, 88% ріпаку в США і
94% у Канаді трансгенні (Levenko, 2011). Трансгенний рослинний організм – це один з видів
генетично модифікованих організмів (ГМО) рослин-
ного походження. В Україні діє закон (Zakon Ukrainy),
що регулює безпеку обігу ГМО та продукції. ваних заходів для забезпечення належного рівня захи-
сту в галузі використання сучасних біотехнологій і,
зокрема, ГМО. У пер-
шій статті цього закону дано визначення: генетично
модифікований організм, живий змінений організм
(ГМО) – будь-який організм, у якому генетичний мате-
ріал був змінений за допомогою штучних прийомів
переносу генів, які не відбуваються у природних умовах,
а саме: рекомбінантними методами, які передбачають
формування нових комбінацій генетичного матеріалу
шляхом внесення молекул нуклеїнової кислоти (вироб-
лених у будь-який спосіб зовні організму) в будь-який
вірус, бактеріальний плазмід або іншу векторну систему
та їх включення до організму-господаря, в якому вони
зазвичай не зустрічаються, однак здатні на тривале роз-
множення; методами, які передбачають безпосереднє
введення в організм спадкового матеріалу, підготовле-
ного ззовні організму, включаючи мікроін’єкції, макроі-
н'єкції та мікроінкапсуляції; злиття клітин (у тому числі
злиття протоплазми) або методами гібридизації, коли
живі клітини з новими комбінаціями генетичного мате-
ріалу формуються шляхом злиття двох або більше клі-
тин у спосіб, який не реалізується за природних обста-
вин. у
р
(
,
)
Набагато обережніше ставлення до ГМО у Європі. Важливим принципом європейського права є принцип
перестороги (Malus, 2016). Країни Європейського
союзу неодноразово скасовували дозволи та піддава-
ли забороні використання тих чи інших ГМО, зокрема
трансгенних рослин. У США поширена думка, що з
боку ЄС діє неоголошений мораторій на імпорт про-
дуктів, у створенні яких було використано методи
біотехнології – відкриті суперечки з цих питань роз-
почались фактично у 2003 році. Румунія, що вступила
у ЄС 2007 року, зазнала збитків, оскільки була зму-
шена відмовитись від вирощування ГМ сої, стійкої до
дії деяких гербіцидів (Bashuk, 2017). З 2003 року ЄС
запровадив, ймовірно, найбільш жорсткі правила
щодо регуляції обігу ГМО (Davison, 2010). Серед
країн ЄС найбільше поширення у сільському госпо-
дарстві набули ГМО у Франції, Італії, Іспанії та Вели-
кобританії (Khmara, 2017). Популярністю тут корис-
туються так звані арктичні яблука, що не коричневі-
ють на зрізі, стійкі до гниття помідори Flavr Savr,
картопля Amflora, що продукує крохмаль певного
типу, “блакитна” троянда Suntory тощо. Серед ГМО
рослин, що були одними з перших, варто згадати тю-
тюн – ще 1986 року у Франції набув поширення стій-
кий до хвороб та шкідників тютюн. У ЄС дозволено
до використання також два види бавовни і три види
кукурудзи, проте у 2015 році 19 держав-членів ЄС
звернулися до Європейської комісії з проханням про
заборону на їх території ГМ кукурудзи MON 810 (а
також картоплі Amflora). Серед держав, що зверну- Поширення продуктів з ГМО у світі та в Україні
зокрема перетворюється у реальність. Перспективи трансгенних рослин у агросфері Львівщини Р.П. Параняк, Б.М. Калин, Б.В. Гутий Р.П. Параняк, Б.М. Калин, Б.В. Гутий Р.П. Параняк, Б.М. Калин, Б.В. Гутий Львівський національний університет ветеринарної медицини та біотехнологій імені С.З. Ґжицького
Л
і
У
ї Львівський національний університет ветеринарної медицини та біотехнологій імені С.З. Ґжицького
м. Львів, Україна У роботі розглянуто стан та перспективи використання трансгенних рослин у сільському господарстві Львівської області. Трансгенні рослини є різновидом генетично модифікованих організмів, одержаних біотехнологічними методами із використанням
транс-переносу (між неспорідненими видами) генів. Протягом останніх років такі культури набули значного поширення у світі;
переважна більшість площ сої, кукурудзи та значна інших культур засіяні ГМ сортами. У країнах ЄС відсутнє широке сприйнят-
тя таких культур та є суттєві обмеження на їх поширення. У ЄС дозволено близько 70 видів ГМО, у тому числі чимало декора-
тивних продуктів (квіти), та близько 58 ГМО для використання у харчових продуктах та кормах. Загалом у світі зареєстровано і
допущено до промислового виробництва продуктів харчування та кормів 168 ліній генетично модифікованих рослин: кукурудзи,
ріпаку, бавовни, картоплі, сої, рису, томатів, пшениці та інших. У нашій країні також є певний досвід отримання трансгенних
організмів. За неофіційними даними, в Україні трансгенні 60–70% сої, 10–20% кукурудзи, 5% – ріпаку. У найближчій перспективі
Україні слід виробити однозначну концепцію ставлення до ГМО, відсутність якої ускладнює планування напрямів діяльності агро-
сектору. У Львівській області є перспективи вирощування ГМ сортів сої, кукурудзи та соняшнику. Вирощування ГМ соняшнику 54 Науковий вісник ЛНУВМБ імені С.З. Ґжицького. Серія: Сільськогосподарські науки, 2019, т 21, № 90 порівняно зі звичайним буде мати обмежені конкурентні переваги. З інших культур, на виробництві яких спеціалізується область,
можна відмітити картоплю, ГМ сорти якої використовують переважно для технічних потреб, та пшеницю, ГМ сорти якої ще
не були достатньо апробовані. На світовому ринку існує попит на генетично модифіковані культури, за оцінками близько 68%
українських агрохолдингів заявляють про бажання та можливості вирощувати ГМ культури за їх легалізації в Україні. порівняно зі звичайним буде мати обмежені конкурентні переваги. З інших культур, на виробництві яких спеціалізується область,
можна відмітити картоплю, ГМ сорти якої використовують переважно для технічних потреб, та пшеницю, ГМ сорти якої ще
не були достатньо апробовані. На світовому ринку існує попит на генетично модифіковані культури, за оцінками близько 68%
українських агрохолдингів заявляють про бажання та можливості вирощувати ГМ культури за їх легалізації в Україні. Ключові слова: аграрний сектор, трансгенний рослинний організм, продукція рослинництва, Львівська область. Scientific Messenger LNUVMB. Series: Agricultural sciences, 2019, vol. 21, no 90
55 ваних заходів для забезпечення належного рівня захи-
сту в галузі використання сучасних біотехнологій і,
зокрема, ГМО. Тому, в кон-
тексті розвитку вітчизняного аграрного сектору важ-
ливо розуміти межі, напрямки та перспективи засто-
сування культур, геном яких модифіковано, для виро-
бництва конкурентоспроможної продукції в умовах
виходу на світові ринки. У даній роботі розглянуто потенційний економіч-
ний ефект та ризики для довкілля у екосистемах різного
типу у Львівської області за умови запровадження
трансгенних рослинних організмів. Аналіз останніх досліджень і публікацій. Україна
приєдналася до Картахенського протоколу про біобе-
зпеку Конвенції про біорізноманіття (від 12/09/2002),
що декларує сприяння забезпеченню належного рівня
захисту від потенційних ризиків, пов’язаних із поши-
ренням ГМО (Oblat, 2014), засвідчивши позицію щодо
підтримки нею необхідності застосування скоордино- Scientific Messenger LNUVMB. Series: Agricultural sciences, 2019, vol. 21, no 90 Scientific Messenger LNUVMB. Series: Agricultural sciences, 2019, vol. 21, no 90 55 Науковий вісник ЛНУВМБ імені С.З. Ґжицького. Серія: Сільськогосподарські науки, 2019, т 21, № 90 лись, – наш сусід Польща. Аргументом на користь
заборони є постанова уряду 2013 року та дані про те,
що пилок генномодифікованої кукурудзи завдає шко-
ди медоносним рослинам і може викликати важкі
наслідки для бджолярства. Цікавою є динаміка основних культур: у 1990 році
кормові культури займали 44% й посідали перше
місце, оскільки однією з причин була потреба підтри-
мки тваринництва. Зернові займали 35%, картопля 9%
і 8% – інші культури з перелічених на рис.1. З плином
часу частка зернових зростала (максимум 50% площ у
2008 році), а кормових – стрімко зменшувалась. Для
картоплі є характерним плавне збільшення до 18% у
2007 та 2010 роках. З 2010 року можна помітити стрі-
мкий ріст інших культур. Динаміку проілюстровано
на рис. 2. лись, – наш сусід Польща. Аргументом на користь
заборони є постанова уряду 2013 року та дані про те,
що пилок генномодифікованої кукурудзи завдає шко-
ди медоносним рослинам і може викликати важкі
наслідки для бджолярства. Загалом у ЄС дозволено близько 70 видів ГМО, в
тому числі чимало декоративних продуктів (квіти) та
близько 58 ГМО для використання у харчових проду-
ктах та кормах (Malus, 2016). Згідно ж з базою даних
AGBIOS (Bashuk, 2017), у світі зареєстровано і допу-
щено до промислового виробництва їжі та кормів 168
ліній генетично модифікованих рослин: кукурудзи,
ріпаку, бавовни, картоплі, сої, рису, томатів, пшениці
та ін. Рис. 1. Посівні площі основних
сільськогосподарських культур у Львівській області в
2018 році (культура; площа, тис. га; відсоток від усіх
культур) (Baza danykh Holovnoho upravlinnia statystyky
u Lvivskii oblasti) Треба розрізняти ГМО та генетично модифікова-
ний продукт – продукт, вироблений за допомогою чи
з використанням ГМО. ваних заходів для забезпечення належного рівня захи-
сту в галузі використання сучасних біотехнологій і,
зокрема, ГМО. У ході приготування продукту
часто генетична інформація втрачається. Щобільше –
прихильники генетично модифікованої продукції
стверджують, що у випадку продуктів харчування
чужорідні вставки повністю руйнуються в системі
травлення тварин і людини (Bashuk, 2017). Результати та їх обговорення ту) та Vistive Gold або MON 87705 – із поліпшеним
синтезом жирних кислот. Загалом ж відомими вважа-
ють 13 ліній сої. Варто зазначити, що стійкість до
гліфосату (N-(phosphonomethyl)glycine) означає лиш
те, що на полі із такою соєю можна більш ефективно
позбуватись бур’янів, застосовуючи гербіцид, що є
ймовірним канцерогеном для людини (категорія небе-
зпеки “2А”). Рис. 3. Динаміка структури посівних площ інших
сільськогосподарських культур у Львівській області
(побудовано на основі даних (Baza danykh
Holovnoho upravlinnia statystyky u Lvivskii oblasti)) Всього відомо понад 22 лінії ГМ картоплі, до най-
відоміших можна зачислити генетично модифікова-
ний сорт картоплі, розроблений BASF Plant Science й
відомий як Amflora (також як EH92-527-1). Рослини
картоплі цього сорту виробляють чистий амілопекти-
новий крохмаль. Цей сорт розглядають як технічний і
його дозволено вирощувати для технічних потреб у
деяких країнах ЄС, наприклад Німеччині. Також було
отримано сорти, стійкі до нематоди, проте лаборатор-
ні дослідження показали, що така картопля спричиняє
зниження імунітету та численні захворювання внут-
рішніх органів у піддослідних щурів. Деякі види ГМ
картоплі доволі алергенні. Тому в більшості держав
ЄС вирощування ГМ-картоплі заборонено. Рис. 3. Динаміка структури посівних площ інших
сільськогосподарських культур у Львівській області
(побудовано на основі даних (Baza danykh
Holovnoho upravlinnia statystyky u Lvivskii oblasti)) Рис. 3. Динаміка структури посівних площ інших
сільськогосподарських культур у Львівській області
(побудовано на основі даних (Baza danykh
Holovnoho upravlinnia statystyky u Lvivskii oblasti)) Як бачимо, найбільше зростання останніми роками
демонструють соя та соняшник, дещо раніше відбу-
лось зростання частки ріпаку, частка якого за останні
8 років змінюється мало. Зменшуються площі під
буряком. На ринку України пропонують насіння трансген-
ного гібриду олійного соняшнику CRESTON FS 799,
що заявлений як стійкий до хвороб, шкідливих комах
та стресових умов. Його обробляють гербіцидом сис-
темної дії імідазолінової групи Євро-Лайтінг, Євро-
Ленд або Євро-Ланг р.к. для знищення широкого
спектру однорічних злакових, багаторічних бур’янів,
дводольних бур’янів. Розроблений канадським Хол-
дингом SERTIS і американським хімічним концерном
UNION CARBIDE в 2016 р. гібрид є самозапильний,
декларується його стійкість до кислотних дощів і
короткочасних перепадів температури у межах від
–12 до +60 ºС. На основі аналізу графіків на рис. 2 і 3 можна
стверджувати, що у Львівській області у найближчій
перспективі основними культурами залишаються
зернові (це на 45% пшениця озима, 31% кукурудза на
зерно, 7% ячмінь озимий, по 6% пшениця та ячмінь
ярі і 4% овес), соя, соняшник та картопля. Розглянемо
перспективи залучення ГМ сортів даних культур. Генетично модифікована пшениця є перспектив-
ним напрямом досліджень у біотехнології. Результати та їх обговорення Рис. 1. Посівні площі основних У Львівській області активно розвивається рослин-
ництво, причому останніми роками спостерігаємо ста-
ле зростання врожайності основних сільськогосподар-
ських культур (Baza danykh Holovnoho upravlinnia
statystyky u Lvivskii oblasti). Важливим показником
спеціалізації є структура площ, зайнятих основними
культурами. До таких можна віднести зернові, кормові
та картоплю. Інші культури займають менш як 15%
кожна і від 6 до 28% (у різні роки) в сукупності. Ста-
ном на 2018 рік у структурі посівних площ сільського-
сподарських культур 44%, або 295,9 тис. га займали
зернові (рис. 1), далі у порядку спадання площ картоп-
ля, кормові культури, ріпак і кольза (ріпак ярий), соя,
соняшник, овочеві культури, буряк цукровий. Рис. 1. Посівні площі основних
сільськогосподарських культур у Львівській області в
2018 році (культура; площа, тис. га; відсоток від усіх
культур) (Baza danykh Holovnoho upravlinnia statystyky
u Lvivskii oblasti) У 2012 році категорія “інші” випередила картоп-
лю, а з 2013 року й кормові культури і продовжує
стрімко зростати. Якщо лінійна тенденція останніх
п’яти років збережеться, то до 2022 року ця категорія
випередить зернові, а тому слід уважно вивчити куль-
тури, площі під якими зростають найшвидше. S i
tifi M
LNUVMB S i
A
i
lt
l
i
2019
l 21
90
Рис. 2. Динаміка структури посівних площ основних сільськогосподарських культур у Львівській області
(побудовано на основі даних (Baza danykh Holovnoho upravlinnia statystyky u Lvivskii oblasti) Рис. 2. Динаміка структури посівних площ основних сільськогосподарських культур у Львівській
(побудовано на основі даних (Baza danykh Holovnoho upravlinnia statystyky u Lvivskii oblast Рис. 2. Динаміка структури посівних площ основних сільськогосподарських культур у Львівській області
(побудовано на основі даних (Baza danykh Holovnoho upravlinnia statystyky u Lvivskii oblasti) аміка структури посівних площ основних сільськогосподарських культур у Львівській області
удовано на основі даних (Baza danykh Holovnoho upravlinnia statystyky u Lvivskii oblasti) Scientific Messenger LNUVMB. Series: Agricultural sciences, 2019, vol. 21, no 90 56 Науковий вісник ЛНУВМБ імені С.З. Ґжицького. Серія: Сільськогосподарські науки, 2019, т 21, № 90 ту) та Vistive Gold або MON 87705 – із поліпшеним
синтезом жирних кислот. Загалом ж відомими вважа-
ють 13 ліній сої. Варто зазначити, що стійкість до
гліфосату (N-(phosphonomethyl)glycine) означає лиш
те, що на полі із такою соєю можна більш ефективно
позбуватись бур’янів, застосовуючи гербіцид, що є
ймовірним канцерогеном для людини (категорія небе-
зпеки “2А”). Всього у категорії “інші” об’єднано 5 культур: бу-
ряк цукровий, соняшник, соя, ріпак та овочеві культу-
ри. Їхню динаміку у вигляді графіка із накопиченням
показано на рис. 3. Scientific Messenger LNUVMB. Series: Agricultural sciences, 2019, vol. 21, no 90
57 Результати та їх обговорення Станом на
2015 року жодна ГМ-пшениця ще не вирощувалась на
комерційній основі, хоча було проведено уже багато
польових випробувань. Пшениця сама по собі є між-
видовим
гібридом
диких
предків
(Triticum
monococcum, Aegilops speltoides і Aegilops tauschii –
усі диплоїдні трави). До 2013 р. у Європі було прове-
дено 34 польових випробування пшениці, а 419 – у
США. Бажані модифікації включають ті, які створю-
ють стійкість до гербіцидів, стійкість до комах і до
грибкових патогенів (особливо Fusarium) та вірусів,
толерантність до посухи та стійкість до солоності ґрун-
ту й надмірного тепла, намагаються знизити вміст
глютеніну, поліпшити харчову цінність (більш висо-
кий вміст білка, підвищення термостійкості ферменту
фітази, збільшення вмісту водорозчинних харчових
волокон, збільшення вмісту лізину), поліпшуються
якості для використання як біопаливної сировини,
застосування до виробництва лікарських засобів й
збільшення врожайності. У нашій країні також є певний досвід отримання
трансгенних організмів. Так, за даними (Levenko, 2011),
в Інституті клітинної біології та генетичної інженерії
НАН України виведено: - горох, стійкий до гербіцидів фосфінотрицину,
персюїту; - цукровий буряк 0-типу, стійкий до фосфінотри-
цину; у
- стійкий до фосфінотрицину ріпак; - стійкий до фосфінотрицину ріпак; - салат, тютюн, моркву з людським геном інтер-
ферону альфа-2b; - салат, цикорій з антигеном ESAT6 бактерії
Mycobacterium tuberculosis, що викликає туберкульоз
(зауважимо, що вакцини і сироватки з рослин – один з
найперспективніших напрямів біотехнології у світі). В Інституті фізіології рослин і генетики та
Нацiональному ботанiчному саду отримано: - сою, цукровий буряк, картоплю, гречку, стійкі до
гербіциду гліфосату; - виноград, стійкий до фосфінотрицину і бактеріа-
льного раку; Соя – культура, що зазнала чи не найбільше різно-
видів генних модифікацій із поширенням відповідних
сортів у землеробній практиці – у США з 1998 року;
станом на 2014 р. в усьому світі офіційно близько
82% посівів сої належали до ГМО. До найбільш попу-
лярних модифікацій сої належать: Roundup ready
soybean – стійка до дії гербіцидів (передусім гліфоса- - ріпак,
олійну
редьку
з
геном
пірролін-5-
карбоксилатсинтетази Medicago truncatula, що підви-
щує вміст проліну, Brassica napus, Raphantus sativus з
супресором гена проліндегідрогенази з підвищеною
стійкістю до абіотичних стресів. Scientific Messenger LNUVMB. Series: Agricultural sciences, 2019, vol. 21, no 90 57 Науковий вісник ЛНУВМБ імені С.З. Ґжицького. Серія: Сільськогосподарські науки, 2019, т 21, № 90 zdobuttia nauk. stupenia kand. ekon. nauk: spets. 08.00.02
“Svitove
hospodarstvo
i
mizhnarodni
ekonomichni vidnosyny (in Ukrainian). За неофіційними даними, в Україні трансгенні 60–
70% сої, 10–20% кукурудзи, 5% – ріпаку (Levenko,
2011). Результати та їх обговорення Існує декілька робіт, у яких наводять результа-
ти досліджень вмісту трансгенних рослин і продуктів
в Україні та Львівській області зокрема, наприклад
(Kushnir, 2015). Зазвичай інтерес дослідників полягає
у виявленні трансгенних продуктів у сировині та про-
дукції рослинництва. Використовують тест-системи
переважно іноземного виробництва для виявлення
маркерів, присутність яких характерна для ГМ рос-
лин. Дослідження зразків на наявність ГМО прово-
дять зокрема методом полімеразної ланцюгової реак-
ції в режимі реального часу згідно з ДСТУ ISO
21569:2008, ДСТУ ISO 21570:2008. Baza danykh Holovnoho upravlinnia statystyky u
Lvivskii
oblasti
[Elektronnyi
resurs]. http://database.ukrcensus.gov.ua/MULT/Dialog/statfil
e_c.asp. Davison, J. (2010). GM plants: Science, politics and EC
regulations. Plant
Science,
178(2),
94–98. doi: 10.1016/j.plantsci.2009.12.005. Khmara,
M.P. (2017). Rol
henno-modyfikovanykh
produktiv v suchasnomu sviti. Mizhnarodni vidnosyny,
Seriia “Ekonomichni nauky”, 9. Elektronnyi resurs. http://journals.iir.kiev.ua/index.php/ec_n/article/view/3
121 (in Ukrainian). Оскільки на світовому ринку існує попит на гене-
тично модифіковані культури, за оцінками близько
68% українських агрохолдингів заявляють про ба-
жання та можливості вирощувати ГМ культури за їх
легалізації в Україні. Kushnir, H.V. (2015). Doslidzhennia roslynnoi syrovyny
na naiavnist henetychno modyfikovanykh dzherel. Naukovo-tekhnichnyi
biuleten
Instytutu
biolohii
tvaryn i Derzhavnoho naukovo-doslidnoho kon-
trolnoho
instytutu
vetpreparativ
ta
kormovykh
dobavok, 16(1), 100–103 (in Ukrainian). Висновки Levenko, B. (2011). Transhenni kultury u sviti ta Ukraini. Visnyk Natsionalnoi akademii nauk Ukrainy, 9, 31–
40. http://nbuv.gov.ua/UJRN/vnanu_2011_9_5
(in
Ukrainian). У світі стрімко поширюється використання ГМО у
сільському господарстві. Разом із тим ставлення до
ГМО у ЄС та США сильно відрізняється, а тому серед
аргументів проти розширення використання цих орга-
нізмів поряд із екологічними та економічними можуть
бути політичні. Malus, A.I. (2016). Derzhavne rehuliuvannia obihu HMO
v
Ukraini,
YeS
ta
SShA. Pravovi
zasady
funktsionuvannia
publichnoi
vlady
shchodo
zabezpechennia intelektualnoho rozvytku ta bezpeky
suspilstva:
materialy
Mizhnarodnoi
naukovo-
praktychnoi konferentsii, Sumy, 19–20 travnia 2016 r. Sumy: SumDU, 86–89 (in Ukrainian). у
У Львівській області основні культури в сільському
господарстві становлять зернові (озима пшениця, куку-
рудза), картопля, стрімко зростає виробництво сої та
соняшника. У світовій практиці поширені ГМ сорти сої
та кукурудзи. На ринку насіння України пропонують
стійкий до погодних умов олійний соняшник, проте
погодні умови тут сприятливі й суттєвої переваги він
не матиме. З ГМ сортів картоплі безпечними вважають
сорти технічного призначення (для виробництва крох-
малю), ГМ сорти пшениці у світовій практиці наразі не
були достатньо апробовані. Таким чином, найбільш
перспективними ГМ культурами для сільського госпо-
дарства Львівщини є соя та кукурудза. У Львівській області основні культури в сільському
господарстві становлять зернові (озима пшениця, куку-
рудза), картопля, стрімко зростає виробництво сої та
соняшника. У світовій практиці поширені ГМ сорти сої
та кукурудзи. На ринку насіння України пропонують
стійкий до погодних умов олійний соняшник, проте
погодні умови тут сприятливі й суттєвої переваги він
не матиме. З ГМ сортів картоплі безпечними вважають
сорти технічного призначення (для виробництва крох-
малю), ГМ сорти пшениці у світовій практиці наразі не
були достатньо апробовані. Таким чином, найбільш
перспективними ГМ культурами для сільського госпо-
дарства Львівщини є соя та кукурудза. Oblat, R.V. (2014). Monitorynh produktiv kharchuvannia
ta silskohospodarskoi syrovyny v Ukraini na vmist
henetychno modyfikovanykh inhrediientiv. Visnyk
ahrarnoi nauky, 1, 59–63. http://nbuv.gov.ua/UJRN/
vaan_2014_1_16 (in Ukrainian). Rudyshyn, S.D. (2011). Henetychno modyfikovani
roslyny: problemy i perspektyvy vykorystannia. Nauka
ta
innovatsii,
7(6),
5–13. http://dspace.nbuv.gov.ua/handle/123456789/115878
(in Ukrainian). Національний науковий потенціал достатній для
проведення досліджень ГМО, проте у законодавстві
наразі не вироблено єдиної концепції ставлення до
цих організмів та продуктів на їх основі. Zakon Ukrainy “Pro derzhavnu systemu biobezpeky pry
stvorenni,
vyprobuvanni,
transportuvanni
ta
vykorystanni
henetychno
modyfikovanykh
orhanizmiv”
vid
31.05.2007
1103-V. Rezhym
dostupu:
http://zakon5.rada.gov.ua/laws/show/1103-
16 (in Ukrainian). Bashuk, V.V. (2017). Formuvannia svitovoho rynku
henetychno modyfikovanykh produktiv: dys. na Scientific Messenger LNUVMB. Series: Agricultural sciences, 2019, vol. 21, no 90
58 References References Bashuk, V.V. (2017). Formuvannia svitovoho rynku
henetychno modyfikovanykh produktiv: dys. na Scientific Messenger LNUVMB. Series: Agricultural sciences, 2019, vol. 21, no 90
58
|
https://openalex.org/W1966032796
|
https://www.scielo.br/j/rap/a/dRQjbqXY9ZsKV4dTBPkKmQJ/?lang=pt&format=pdf
|
Portuguese
| null |
Construindo o perfil do jovem empreendedor brasileiro: relevância para a formulação e implementação de políticas de estímulo ao empreendedorismo
|
Revista de administração pública
| 2,007
|
cc-by
| 3,580
|
* Coordenadores do Programa de Estudos Avançados em Pequenos Negócios, Empreendedorismo,
Acesso ao Crédito e Meios de Pagamento (Small Business), da Escola Brasileira de Administra-
ção Pública e de Empresas da Fundação Getulio Vargas (Ebape/FGV). Endereço: Praia de
Botafogo, 190, 5o andar — CEP 22250-900, Rio de Janeiro, RJ, Brasil. E-mail: small@fgv.br.
** Doutora em engenharia de produção (Coppe/UFRJ). E-mail: deborah.zouain@fgv.br.
*** Doutora em educação, mestre em administração. Professora titular da FGV nos cursos de
doutorado e mestrado. Endereço: Praia de Botafogo, 190, sala 534 — CEP 22250-900, Rio de
Janeiro, RJ, Brasil. E-mail: fbayma@fgv.br.
**** Doutorando em políticas públicas e formação humana (PPFH/Uerj). E-mail: francisco.
barone@fgv.br. Construindo o perfil do jovem empreendedor brasileiro:
relevância para a formulação e implementação de
políticas de estímulo ao empreendedorismo Deborah Moraes Zouain**
Fátima Bayma de Oliveira***
Francisco Marcelo Barone**** Deborah Moraes Zouain**
Fátima Bayma de Oliveira***
Francisco Marcelo Barone**** SUMÁRIO: 1. Introdução; 2. Escopo e metodologia da pesquisa; 3. Análise
de dados; 4. Considerações finais. Coordenação: Deborah Moraes Zouain*
Francisco Marcelo Barone*
............ “Espaço dedicado à divulgação de estudos e pesquisas sobre o conceito de small
business: micro, pequenas e médias empresas (MPMEs); empreendedorismo; jovens
empresários; acesso ao crédito; meios de pagamento; e responsabilidade social.”
................................................................................................................................ .................... 1. Introdução A criação de um ambiente favorável ao desenvolvimento de negócios no Brasil
passa pelo estímulo ao empreendedorismo. Em um país onde 56,20% dos RAP Rio de Janeiro 41(4):797-807, Jul./Ago. 2007 798 Construindo o Perfil do Jovem Empreendedor Construindo o Perfil do Jovem Empreendedor empreendedores iniciais1 são jovens (faixa etária de 18 a 34 anos), de acordo
com o Global Entrepreneurship Monitor de 2006, conhecer o perfil desses jo-
vens é de fundamental importância para o balizamento dos policy makers, em
termos de formulação e implantação de novas políticas públicas de estímulo
ao empreendedorismo. O ponto de partida deste artigo foi a constatação da carência de pesqui-
sas especificamente voltadas às características socioeconômicas dos jovens
empreendedores e seu comportamento empreendedor. A única pesquisa na-
cional voltada ao segmento das microempresas é a Economia Informal Urbana
(Ecinf), do IBGE. Entretanto, ela sofre duas limitações: destina-se ao segmen-
to informal urbano e contempla em sua amostra apenas empreendimentos
com até cinco empregados. Além disso, não há um desenho amostral intencio-
nal voltado a jovens empreendedores. O perfil do jovem empreendedor2 é evidenciado com uma pesquisa es-
pecialmente desenvolvida para este fim, voltada ao conhecimento de suas ca-
racterísticas socioeconômicas, do momento de negócio, das dificuldades
enfrentadas na gestão de seus empreendimentos, e por último, das expectati-
vas em relação aos seus negócios para o futuro. Nos últimos anos é crescente o número de jovens entrando no mercado
de trabalho, e sem condições de serem absorvidos pelas grandes empresas,
essa tendência, segundo projeções, continuará crescente no decorrer do pri-
meiro terço deste século. Desde 2000, o Brasil alcançou a conhecida janela de
oportunidade demográfica3 e irá permanecer nela pelos próximos 30 anos. 2 O corte etário para jovem empreendedor aplicado na pesquisa foi 18 a 34 anos. Ademais
nesta pesquisa não houve limitação quanto ao número de empregados do empreendimento. O
foco era o jovem empreendedor, sendo ele porventura micro, pequeno, médio ou grande em-
pregador. 1 Empreendedores iniciais são aqueles que estão à frente de negócios com até 42 meses de vida
(Global Entrepreneurship Monitor, 2006:36). (Global Entrepreneurship Monitor, 2006:36).
2 O corte etário para jovem empreendedor aplicado na pesquisa foi 18 a 34 anos. Ademais,
nesta pesquisa não houve limitação quanto ao número de empregados do empreendimento. O
foco era o jovem empreendedor, sendo ele porventura micro, pequeno, médio ou grande em-
pregador. 1. Introdução p
g
3 Janela de oportunidade demográfica consiste em uma estrutura demográfica com as seguintes
características específicas: p
g
3 Janela de oportunidade demográfica consiste em uma estrutura demográfica com as seguintes
características específicas: ®a maioria da população encontra-se na faixa etária entre 15 e 64 anos (a população econo-
micamente ativa); ®a maioria da população encontra-se na faixa etária entre 15 e 64 anos (a população econo-
micamente ativa); ®a taxa de crescimento populacional cai fortemente (no Brasil, de 2,89% a.a. entre 1960 e
1970 para 1,64% a.a. entre 1991 e 2000); ®a razão de dependência cai (representa a população economicamente ativa (15-64 anos)
dividida pela população jovem (0-14 anos) mais a população idosa (65 anos ou mais). Um
decréscimo na razão de dependência indica que uma maior população estará apta à ativida-
de economicamente ativa, ou seja, ao trabalho e, então, há um maior potencial de geração
de riqueza e renda per capita (no Brasil, caiu de 63,5% em 1993 para 51,1% em 2003); RAP Rio de Janeiro 41(4):797-807, Jul./Ago. 2007 799 Small Business Através do Panóptico Dependendo das políticas socioeconômicas desenvolvidas e implemen-
tadas nos primeiros 20 anos deste século, a janela de oportunidade demográfica
pode vir a ser uma grande oportunidade de desenvolvimento e de melhoria do
bem-estar ou representar um grande problema socioeconômico futuro. Em
outras palavras, o conjunto das políticas escolhidas propiciará à população
economicamente ativa brasileira: gerar maiores patamares de renda, riqueza e
poupança nos próximos anos; e, também, reconfigurar a nossa estrutura
socioeconômica futura em termos de distribuição de ativos (educação, pro-
priedade, capital etc.) e de bem-estar (Estrella, 2007). É neste processo que o Brasil se encontra, o que torna o empreendedo-
rismo, especialmente com foco no jovem empreendedor, um tema crucial na
agenda presente do desenvolvimento socioeconômico brasileiro, pois afeta
diretamente o processo de inclusão dos jovens no mercado de trabalho. ®a percentagem de idosos é crescente, mas menos que em países com estruturas populacionais
envelhecidas;
®aumenta a taxa de expectativa de vida e caem as taxas de fecundidade e mortalidade.
4 Entidade de âmbito nacional, presente nos 27 estados da federação e no Distrito Federal,
representando mais de 15 mil jovens empresários, que tem por finalidade estimular o
empreendedorismo entre os jovens brasileiros com três motes principais: orientação e capacitação
profissional, representatividade e troca de experiências por uma complexa rede de relaciona-
mentos. ®aumenta a taxa de expectativa de vida e caem as taxas de fecundidade e mortalidade.
4 Entidade de âmbito nacional, presente nos 27 estados da federação e no Distrito Federal,
representando mais de 15 mil jovens empresários, que tem por finalidade estimular o
empreendedorismo entre os jovens brasileiros com três motes principais: orientação e capacitação
profissional, representatividade e troca de experiências por uma complexa rede de relaciona-
mentos. ®aumenta a taxa de expectativa de vida e caem as taxas de fecundidade e mortalidade. 3. Análise de dados Os dados foram trabalhados a partir do corte analítico fundamental entre as-
sociados e não-associados da Conaje. Essa segmentação objetivava conhecer
melhor o perfil do jovem empreendedor, como também observar possíveis di-
ferenças entre os que possuem um viés associativista e os que não buscam o
associativismo como meio de defesa de seus interesses. A análise dos resultados obtidos está apresentada a seguir para os se-
guintes tópicos: caracterização socioeconômica do jovem empreendedor; mo-
mento, ambiente do negócio e dificuldades enfrentadas; tecnologia, finanças e
perspectivas futuras. 2. Escopo e metodologia da pesquisa Buscando definir o perfil do jovem empreendedor, foi desenvolvida uma pes-
quisa aleatória, exploratória e descritiva, utilizando como base de coleta de
dados movimentos estaduais de jovens empresários filiados à Confederação
Nacional dos Jovens Empresários4 (Conaje). O instrumento de coleta desenvolvido é um survey estruturado. Como
este seria aplicado em locais distintos, por pessoas diferentes, objetivou-se
minimizar, ao máximo, os erros de coleta de dados. Dentro dessa perspectiva,
a escolha do survey foi decorrente de suas especificidades: o entrevistador aplica
a mesma série de perguntas padronizadas, ou seja, perguntas detalhadas apli-
cadas a todos os entrevistados, de maneira idêntica, sem alteração de ordem
ou de formulação e codificadas de acordo com regras preestabelecidas, visan-
do minimizar os efeitos da iteração entrevistador-entrevistado (Patton, 2004;
Fontana e Frey, 2000; Marconi e Lakatos, 1999). nta a taxa de expectativa de vida e caem as taxas de fecundidade e mortalidade. 4 Entidade de âmbito nacional, presente nos 27 estados da federação e no Distrito Federal,
representando mais de 15 mil jovens empresários, que tem por finalidade estimular o
empreendedorismo entre os jovens brasileiros com três motes principais: orientação e capacitação
profissional, representatividade e troca de experiências por uma complexa rede de relaciona-
mentos. RAP Rio de Janeiro 41(4):797-807, Jul./Ago. 2007 800 Construindo o Perfil do Jovem Empreendedor Construindo o Perfil do Jovem Empreendedor A pesquisa foi realizada entre os dias 9 e 28 de março de 2007, nos
estados de Santa Catarina, Ceará, Rio Grande do Norte, Sergipe, Pará, Bahia,
Espírito Santo e no Distrito Federal. Foram disponibilizados questionários pela
internet aos associados da Conaje e enviados aos estados e Distrito Federal
questionários a serem aplicados, sob a coordenação de representantes locais, a
não-associados. Posteriormente, esses questionários foram transcritos para for-
mulário eletrônico próprio. A pesquisa contemplou jovens empreendedores associados e não-asso-
ciados entre 18 e 34 anos e foi dimensionada para ser respondida por 1.500
associados e 330 não-associados. O nível de respostas dependia do comprome-
timento dos respondentes, quando associados, e do empenho dos coordenado-
res locais, quando aplicada a não-associados. Os estados com maiores respostas
totais (associados e não-associados) à pesquisa foram: Sergipe (31,6%), Santa
Catarina (22,8%), Ceará (15,8%) e Distrito Federal (12,8%). Caracterização socioeconômica No corte por gênero, para os associados, foram respondentes homens (90,2%)
e mulheres (9,8%). Entre os não-associados ocorre uma ligeira mudança desse
perfil, com ganhos para as mulheres, que dobram a sua participação: repre-
sentaram 18,8% e os homens, 81,2%. Considerando-se a variável raça/etnia,
70,7% dos associados se declararam brancos e 24,3% se declararam pardos. Os demais eram asiáticos ou indígenas. Entre os não-associados ocorre um
crescimento significativo dos pardos, e, em contrapartida, conseqüente redu-
ção dos brancos: 50% se declararam pardos, 31,2% se declararam brancos e
12,5% amarelos, ou seja, de origem asiática. RAP Rio de Janeiro 41(4):797-807, Jul./Ago. 2007 801 Small Business Através do Panóptico Quando observamos a posição no domicílio, 46,3% dos associados são
chefes do domicílio e 48,7% ocupam a posição no domicílio como filhos ou
enteados. Para os não-associados, 43,7% são chefes e 56,3% são filhos ou en-
teados. A seguir, são evidenciados os dados para renda domiciliar mensal dos
pesquisados. Distribuição da renda domiciliar mensal total ç
Fonte::::: Pesquisa Perfil do Jovem Empreendedor (Small Business/Ebape/FGV, mar. 2007). 100%
90%
80%
70%
60%
50%
40%
30%
20%
10%
0%
58,54
9,76
12,20
14,63
4,88
43,75
12,50
6,25
31,25
6,25
Associados
Não-associados
R$ 701 a R$ 1.750
R$ 5.251 a R$ 7.000
R$ 1.751 a R$ 3.500
acima de R$ 7.001
R$ 3.501 a R$ 5.250 Fonte::::: Pesquisa Perfil do Jovem Empreendedor (Small Business/Ebape/FGV, mar. 2007). Cabe evidenciar que a renda domiciliar mensal total de quase 60% dos
associados é alta para os padrões brasileiros (acima de R$ 7 mil). Observando-
se os não-associados, o percentual daqueles com renda maior que R$ 7 mil cai
para 43,8%, e ocorre um crescimento significativo do percentual daqueles que
possuem uma renda de até R$ 3.500, se comparado ao outro grupo. No quesito escolaridade, 39% dos associados possuem alguma pós-gra-
duação, 34,2% possuem superior completo e 26,8% segundo grau completo. Quando olhamos os não-associados, o contexto educacional se altera comple-
tamente: apenas 31,2% possuem superior completo e os demais, 68,8%, pos-
suem somente segundo grau completo. RAP Rio de Janeiro 41(4):797-807, Jul./Ago. 2007 802 Construindo o Perfil do Jovem Empreendedor Momento e ambiente do negócio e dificuldades enfrentadas Na análise quanto ao ambiente de negócios, é importante pontuar a relação
entre associativismo e a idade do empreendimento. Considerando-se os asso-
ciados que possuem empresas, 46% iniciaram seus negócios há mais de cinco
anos, percentual idêntico para os não-associados. Todavia, enquanto 31,2%
dos associados possuem negócios iniciados há menos de três anos, entre os
não-associados, esse percentual sobe para 46,2%. Nas figuras 2 e 3 observa-
mos a distribuição do setor de atividades predominantes das empresas. F i g u r a 2 Setor de atividade econômica: associados
3,13%
Indústria
65,63%
9,38%
21,88%
Serviços
Comércio
Construção civil
Fonte: Pesquisa Perfil do Jovem Empreendedor (Small Business/Ebape/FGV, mar. 2007). Setor de atividade econômica: associados Setor de atividade econômica: associados Fonte: Pesquisa Perfil do Jovem Empreendedor (Small Business/Ebape/FGV, mar. 2007). F i g u r a 3 F i g u r a 3 Setor de atividade econômica: não-associados Fonte: Pesquisa Perfil do Jovem Empreendedor (Small Business/Ebape/FGV, mar. 2007). 7,69%
69,23%
23,08%
Serviços
Comércio
Construção civil Fonte: Pesquisa Perfil do Jovem Empreendedor (Small Business/Ebape/FGV, mar. 2007). 7,69%
69,23%
23,08%
Serviços
Comércio
Construção civil Fonte: Pesquisa Perfil do Jovem Empreendedor (Small Business/Ebape/FGV, mar. 2007). RAP Rio de Janeiro 41(4):797-807, Jul./Ago. 2007 803 Small Business Através do Panóptico Para o grupo dos associados, 17,1% das empresas possuem apenas um
empregado além do proprietário, 12,2% possuem dois empregados e outros
12,2%, quatro empregados. Apenas 4,9% são por conta própria. Observando-
se o outro grupo, o percentual de por conta própria mais que triplica (18,8%)
e a freqüência de empresas que possuem apenas um empregado além do pro-
prietário alcança 43,7%. No aspecto regularização do negócio e associativismo, alguns fatores se
sobressaem. Quanto a possíveis dificuldades para regularizar o negócio, 3,1%
dos associados sequer tentaram a regularização e 43,8% alegaram encontrar
dificuldades para a regularização. Para o grupo dos não-associados, o percentual
dos que não tentaram regularizar sobe para 15,4% e daqueles que encontra-
ram dificuldades cai para 38,5%. Entretanto, entre todos os respondentes dos
dois grupos que encontraram dificuldades em regularizar o negócio, quase
80% alegaram “grande burocracia envolvida na regularização” como a princi-
pal dificuldade enfrentada. p
Outra correlação forte observada ocorre entre associativismo e consti-
tuição jurídica. Ela é importante, por ser um passo para a formalização do
empreendimento e, com isso, o conseqüente aumento da formalização da eco-
nomia. Nos dois grupos, o percentual de empreendedores com registro de
microempresa encontra-se em torno de 60%. F i g u r a 4 Pergunta: a empresa possui constituição jurídica? Pergunta: a empresa possui constituição jurídica? Fonte: Pesquisa Perfil do Jovem Empreendedor (Small Business/Ebape/FGV, mar. 2007). 100%
90%
80%
70%
60%
50%
40%
30%
20%
10%
0%
96,88
30,77
69,23
Associados
Não-associados
Sim
Não
3,13 Fonte: Pesquisa Perfil do Jovem Empreendedor (Small Business/Ebape/FGV, mar. 2007). RAP Rio de Janeiro 41(4):797-807, Jul./Ago. 2007 RAP Rio de Janeiro 41(4):797-807, Jul./Ago. 2007 804 Construindo o Perfil do Jovem Empreendedor Construindo o Perfil do Jovem Empreendedor No quesito “aderir ao Simples”, é evidente a vantagem dos associados
(65,6%) diante dos não-associados (38,5%). O principal motivo alegado para
não aderir está em que as alíquotas não são vantajosas à empresa: 54,5% para
associados e 62,5% para não-associados. Outro aspecto a ser pontuado é a multicolinearidade entre os diferentes tipos
de associativismo. Há uma correlação clara entre ser associado da Conaje e possuir
algum outro tipo de associação, seja sindicato, órgão de classe ou cooperativa. F i g u r a 5 F i g u r a 5
Pergunta: é filiado a algum sindicato ou órgão de classe,
exceto a Conaje? 100%
90%
80%
70%
60%
50%
40%
30%
20%
10%
0%
39,02
4,88
34,15
81,25
18,75
Associados
Não-associados
Sindicato
21,95
Cooperativa
Órgão de classe
Não-associado a nenhum acima
Fonte: Pesquisa Perfil do Jovem Empreendedor (Small Business/Ebape/FGV, mar. 2007). Fonte: Pesquisa Perfil do Jovem Empreendedor (Small Business/Ebape/FGV, mar. 2007). Entre os associados, 61% apresentam outro tipo de associação, enquan-
to entre os não-associados, 81,2% não apresentam qualquer tipo de associati-
vismo. Considerando-se a motivação para se associar, os associados
apresentaram como resposta mais freqüente, quanto à necessidade de associa-
tivismo (44,4%), “a defesa de seus interesses”. Considerando-se os fatores dificuldade no desenvolvimento dos negóci-
os e assistência técnica recebida, a principal dificuldade encontrada pelos as-
sociados no desenvolvimento de seus negócios ao longo de 2006 foi o alto
nível dos impostos (34,38%), enquanto os não-associados responderam o bai-
xo lucro (23,1%). Quanto ao segundo fator, a assistência técnica mais freqüen- RAP Rio de Janeiro 41(4):797-807, Jul./Ago. 2007 805 Small Business Através do Panóptico te recebida entre os associados foi treinamento (30,4%), seguida de igual
percentual (26,1%) por assistência financeira e jurídica. Para os não-associa-
dos, 80% receberam assistência financeira e, os demais, treinamento. te recebida entre os associados foi treinamento (30,4%), seguida de igual
percentual (26,1%) por assistência financeira e jurídica. Para os não-associa-
dos, 80% receberam assistência financeira e, os demais, treinamento. Tecnologia, finanças e perspectivas futuras Para os empreendimentos dos associados em que o celular consistiu em um
instrumento de aumento da receita, ele trouxe um acréscimo de 30% ou mais
em 35,7% dos casos, igual percentual para um aumento de receita de 10% e
em 28,6% dos casos o aumento foi de 20%. No grupo dos não-associados,
quando o celular proporciona aumento da receita do negócio, este foi de 30%
ou mais, em 60% dos casos. É É relevante ressaltar e, constitui também um fator extremamente positi-
vo em termos de gestão e de produtividade, que todas as empresas dos asso-
ciados estão informatizadas, enquanto 75% das empresas dos não-associados
possuem computadores. Entre os associados, 40% possuem sistema integrado
entre a caixa registradora da empresa e o computador, sendo um importante
indicador de qualidade da gestão contábil-financeira da empresa. Para os não-
associados que possuem computador, esse percentual cai para 33%. F i g u r a 6
Receita total da empresa no ano de 2006
100%
90%
80%
70%
60%
50%
40%
30%
20%
10%
0%
7,32
9,76
4,88
14,63
48,78
6,25
18,75
75,00
Associados
Não-associados
Até R$ 120.000
R$ 240.001 a R$ 400.000
R$ 120.001 a R$ 240.000
R$ 400.001 a R$ 1.200.000
R$ 1.200.001 a R$ 2.400.000
R$ 2.400.001 a R$ 5.000.000
14,63
Fonte: Pesquisa Perfil do Jovem Empreendedor (Small Business/Ebape/FGV, mar. 2007). Receita total da empresa no ano de 2006 Fonte: Pesquisa Perfil do Jovem Empreendedor (Small Business/Ebape/FGV, mar. 2007). RAP Rio de Janeiro 41(4):797-807, Jul./Ago. 2007 RAP Rio de Janeiro 41(4):797-807, Jul./Ago. 2007 806 Construindo o Perfil do Jovem Empreendedor Construindo o Perfil do Jovem Empreendedor Um resultado que chamou a atenção foi que enquanto 41,3% dos asso-
ciados alegaram ter dificuldades para acessar serviços financeiros, esse valor
caiu 10 pontos em relação aos não-associados (31,4%). Esse é um dado que
merece ser melhor investigado em novas pesquisas devido ao fato de que ser
associado, em tese, deveria ser mais vantajoso para acesso a serviços financei-
ros, já que a entidade de classe pode negociar instrumentos financeiros para o
conjunto de seus membros junto ao setor financeiro. Entre os associados, a
principal dificuldade apresentada para acessar serviços financeiros foi o alto
custo das tarifas financeiras (47,1%), enquanto entre os não-associados, a
motivação mais freqüente foi a insuficiência de renda (40%). Considerando-se o instrumento cartão de crédito, apenas cerca de 40%
dos respondentes totais percebem vantagem em possuir um cartão de crédito
empresarial, exclusivo para uso da empresa. Todavia, entre aqueles que vêem
vantagem em possuir o cartão, cerca de 33% consideram que o fator mais
vantajoso é o melhor controle dos gastos da empresa. Essa última percepção é
bastante relevante quando atentamos para a melhoria da gestão contábil-fi-
nanceira dos pequenos e novos negócios. Sabemos que um dos motivos da
mortalidade das empresas é justamente a falta de controle das contas, fator
fortemente alardeado nas pesquisas do Sebrae. O cartão empresarial é um
instrumento importante tanto para o controle das contas do empreendimento,
quanto para a separação entre contas do negócio e contas pessoais e também
quanto à redução de custos de transação financeira, pois reduz sobremaneira
o uso dos boletos bancários. Por fim, no que se refere às perspectivas futuras, 85% dos associados
planejam aumentar o negócio, enquanto entre os não-associados esse número
cai para 62,5%, seguido de 12,5% que pretendem mudar de atividade, mas
continuar independentes. 4. Considerações finais As características já evidenciadas nos três eixos temáticos (caracterização
socioeconômica; momento e ambiente do negócio e dificuldades enfrentadas;
tecnologia, finanças e perspectivas futuras) são de fundamental importância
para a consubstanciação de políticas públicas focadas no segmento das micro
e pequenas empresas, visando o empoderamento do jovem empreendedor e,
por conseqüência, o fortalecimento da economia como um todo. Dentro dessa perspectiva, esta pesquisa levantou um perfil inicial desse
segmento e gerou informações para subsidiar futuras ações, públicas e/ou priva- RAP Rio de Janeiro 41(4):797-807, Jul./Ago. 2007 807 Small Business Através do Panóptico das, que coloquem os jovens empreendedores como atores relevantes no pro-
cesso de desenvolvimento socioeconômico brasileiro. das, que coloquem os jovens empreendedores como atores relevantes no pro-
cesso de desenvolvimento socioeconômico brasileiro. Referências bibliográficas ESTRELLA, J. Regarding microcredit as a policy to reduce inequalities and improve well-
being. 2007. (Versão preliminar da tese de doutorado). FONTANA, A.; FREY, J. H. The interview. From structured questions to negotiated text. In:
DENZIN, N. K.; LINCOLN, Y. S. (Orgs.). Handbook of qualitative research. 2. ed. London:
Sage Editions, 2000. IADB (Inter-American Development Bank). Demography: threat or opportunity? In: IADB
(Org.). Development: beyond economics. Washington, DC: IADB, 2000. IBGE (Instituto Brasileiro de Geografia e Estatística). Economia informal urbana (Ecinf). Rio de Janeiro: IBGE, 2003. ______. Síntese de indicadores sociais 2004. Rio de Janeiro: IBGE, 2005. MARCONI, M. A.; LAKATOS, E. M. Técnicas de pesquisa. São Paulo: Atlas, 1999. MINAYO, M. C. S.; SOUZA, E. R.; ASSIS, S. (Orgs.). Avaliação por triangulação de métodos. Rio de Janeiro: Fiocruz, 2005. PATTON, M. Q. Qualitative research & evaluation methods. London: Sage, 2004. PENN, G. Análise semiótica de imagens paradas. In: BAUER, M.; GASKELL, G. (Orgs.). Pesquisa qualitativa com texto, imagem e som. Petrópolis: Vozes, 2002. SCHLEMM, M. M. Empreendedorismo no Brasil. Curitiba: IBPQ, 2007. RAP Rio de Janeiro 41(4):797-807, Jul./Ago. 2007
|
https://openalex.org/W2056498645
|
https://ruj.uj.edu.pl/xmlui/bitstream/handle/item/132783/sobolewska_podolak_makowska-was_allium_ursinum_2015.pdf?sequence=1&isAllowed=y
|
English
| null |
Allium ursinum: botanical, phytochemical and pharmacological overview
|
Phytochemistry reviews
| 2,013
|
cc-by
| 12,909
|
Phytochem Rev (2015) 14:81–97
DOI 10.1007/s11101-013-9334-0 Phytochem Rev (2015) 14:81–97
DOI 10.1007/s11101-013-9334-0 Allium ursinum: botanical, phytochemical
and pharmacological overview Danuta Sobolewska • Irma Podolak •
Justyna Makowska-Wa˛s Received: 9 May 2013 / Accepted: 18 December 2013 / Published online: 25 December 2013
The Author(s) 2013. This article is published with open access at Springerlink.com folk tales, according to which bears after awaken-
ing from winter hibernation consume this plant to
remove toxins from the body and to regain strength
(Rejewski 1996). Another etymological hypothesis
refers to the ‘‘Ursa major’’, a constellation in the
northern hemisphere, perhaps because A. ursinum
was, to ancient Greeks especially, one of the most
northerly distributed Allium species (Bo¨hling 2008). Contemporary systematics places this plant in the
family Amaryllidaceae (previously in Alliaceae),
subgenus Amerallium Traub, section Arctoprasum
Kirschl. (Friesen et al. 2006; Chase et al. 2009;
Govaerts 2011). Several synonyms are recognized:
Allium nemorale Salisb., A. latifolium Gilib., and
Ophioscorodon ursinum (L.) Wallr. Thanks to its
wide-spread distribution and popularity as edible
and medicinal plant, most modern European lan-
guages have common names for A. ursinum which
are used interchangeably. These are: Ramson or
Bear’s garlic (English); Ba¨rlauch (German); Ail des
ours, Ail sauvage (French); Kyr velde;bq, Xecyor
velde;bq or Xepevia (Russian); Ramslo¨k (Swed-
ish);
Daslook
(Dutch);
Czosnek
niedz´wiedzi
(Polish). Abstract
Ramson—Allium ursinum L. is a medic-
inal and dietary plant species with a long tradition of
use. This mini-review summarizes the current knowl-
edge on the phytochemistry and pharmacological
properties of this valuable plant, with special emphasis
on antimicrobial, cytotoxic, antioxidant, and cardio-
protective effects. Keywords
Allium ursinum Biological
activity Ramson Steroidal saponins Sulfur
compounds D. Sobolewska (&) I. Podolak J. Makowska-Wa˛s
Department of Pharmacognosy, Jagiellonian University,
Medical College, 9 Medyczna Street, Krako´w, Poland
e-mail: dsobolew@cm-uj.krakow.pl Description As far as morphological features are concerned
ramson is a typical representative of the Allium genus. The plant is a bulbiferous, vernal geophyte. Its bulb is
narrow, elongated, about 1.5–6 cm long, surrounded
by the layers of clear skin with only a few fibers at the
base. Sometimes daughter bulbs are formed, what is
important for vegetative reproduction. Contractile
roots start to develop approximately from the age of
three (Eggert 1992; Ernst 1979; Szafer et al. 1988;
Oborny et al. 2011; Macku˚ and Krejcˇa 1989; Cˇ incˇura
et al. 1990). When the soil is soft enough to enable the
roots to dwell deeper and deeper, after 10 years they
can reach down the level even 27 cm lower (Ellenberg
1988). Ramson grows up to the height of 50 cm. The
aerial parts of the plant consist of a triquetrous, erect,
flower stem, solid in cross-section. Atop a stalk, there
is a semispherical umbel-like inflorescence, which
comprises
of
3–30
starry,
snowy-white
flowers
(according to Bła_zewicz-Woz´niak: 13.4–24.0 on
average). They are surrounded by 2–3 spathal bracts
until anthesis. Flower parts are in 6’s sets (Eggert
1992; Ernst 1979; Berger 1960; Szafer et al. 1988;
Rejewski 1996; Macku˚ and Krejcˇa 1989; Cˇ incˇura et al. 1990; Bła_zewicz-Woz´niak et al. 2011). Ramson’s
blooming usually starts in April and ends in the first
half of May. Introduction Allium ursinum L. has been used for centuries in
traditional medicine. However, studies on its compo-
sition and pharmacological activity are fairly recent
and scarce. The aim of the present review was to
summarize the most important aspects related to A. ursinum and provide an outline of phytochemical and
pharmacological properties of this relatively poorly
known plant species of the Allium genus. The species name ‘‘ursinum’’ is of Latin origin,
being derived from ‘‘ursus’’ (bear), and is related to Also, the name ‘‘wild garlic’’ is very often used
in literature with respect to A. ursinum, though it
can be sometimes ambiguous, since it also refers to
other species, like A. vineale or A. canadense, as
well as to plants from the genus Tulbaghia (Defelice
2003; Maine Rare Plant List 2013; Lyantagaye
2011). 12 3 3 Phytochem Rev (2015) 14:81–97 82 the seeds fall onto the ground directly beneath the
capsules, but they seem to be too heavy to be moved by
the wind at a ground level (Oborny et al. 2011; Ernst
1979). So, for a long-distance transport to potential
growing sites the participation of animals or running
water is needed (Eggert 1992). Most of the seeds
remain dormant for 1 or 2 years, however, some
germinate in the course of upcoming winter or spring,
usually from November to March (according to Ernst
it takes place from January to April) (Eggert 1992;
Ernst 1979). A dense carpet of A. ursinum can produce a
large number of seeds annually, even 10,000 seeds/m2
as was recorded in Gottingen Forest (Germany) (Ernst
1979). In Litovelske´ Pomoravı´ (Czech Republic)
floodplain forest, the mean seed production was esti-
mated as 2,692 seeds per m2 (max 5,612 seeds/m2)
(Rychnovska´ and Bedna´rˇ 1998). 123 Geographical distribution and habitat
requirements Allium ursinum is a perennial herbaceous species, of
wide-spread distribution both in Europe and Asia. Although, not growing at high altitudes (beyond
1.900 m) and in the far North (beyond ca 64N), it
can be found on natural stands from the Mediterranean
region to Scandinavia (Oborny et al. 2011). It is also
native to Asia Minor, the Caucasus, and Siberia, up to
the Kamchatka Peninsula (Rola 2012; Madaus 1938;
Oborny et al. 2011; Djurdjevic et al. 2004). The plant develops 2–3 leaves, which are shorter
than stem, smooth, flat, elliptic-lanceolate with a
distinct, well-developed blade, sharpened at the apex,
and gradually narrowed into petiole at the base. The
width of ramson leaves is 20–64 mm (Szafer et al. 1988; Bła_zewicz-Woz´niak and Michowska 2011). A
comparative study on a collection of A. ursinum
specimens from different ecotypes in Poland (Dukla,
Roztocze, Bieszczady) showed that, they differed
significantly in the width of leaf blades, the length of
leaf stalks and flowering stems, the number of flowers
in inflorescences (Bła_zewicz-Woz´niak and Mich-
owska 2011). Two subspecies of A. ursinum are recognized: A. ursinum ssp. ursinum and A. ursinum ssp. ucrainicum. The subdivision is based on the smoothness of pedicel
surface (Karpaviciene 2006). The pedicels of the ssp. ursinum are scabrid, with numerous papillae, and rough,
while the ssp. ucrainicum has smooth pedicels without
papillae (Rola 2012; Farkas et al. 2012). The former is
distributed in western and central Europe, whereas the
latter in eastern and southeastern part of the continent
(Rola 2012; Oborny et al. 2011). The distribution areas
of the two subspecies can overlap what results in the
existence of transitional forms. In Poland, both subspe-
cies along with the intermediate forms, occurring at the
border of the distribution ranges, were recorded (Rola
2012). In West Caucasus and Germany ssp. ucrainicum
is sometimes transplanted into gardens (Hanelt and
Bu¨ttner 2001).Also,cultivationtrialshavebeenmadein
former Czechoslovakia. Allium ursinum regenerates mainly by seeds; veg-
etative regeneration is of minor importance. The seeds
are black, subglobose, 2–3 mm wide, gathered in
trichotomic capsules (Hermy et al. 1999; Sendl 1995). Mean weight per seed is 5.4 ± 0.7 mg. They are shed
in June and July, however shedding time may be
delayed by weather conditions, e.g. a cold spring and
summer, or north-facing aspect (Ernst 1979). Most of 123 123 Phytochem Rev (2015) 14:81–97 83 The habitat preferences are equal for both subspe-
cies (Rola 2012). Chemical constituents Allium ursinum has a distinct garlic-like scent asso-
ciated with the presence of sulfur-containing com-
pounds which are the most characteristic constituents
in Allium plants. Geographical distribution and habitat
requirements Ramson—flourishes best in light and
medium, nutrient-rich, damp, but well-drained soils,
in full shade and semi-shade localities (Szafer and
Zarzycki 1972; Cˇ incˇura et al. 1990; Oborny et al. 2011). Although the plant prefers high air humidity, it
can be also found on shallow, and fairly dry, in
summer months especially, calcareous soil (Eggert
1992). However, both waterlogging and drought are
considered as restricting factors for ramson (Kovacs
2007). Another factor limiting its distribution is the
aluminium concentration in the soil water (Andersson
1993). Under experimental conditions aluminium (at
the concentration of 20 lM) severely restricted ram-
son root elongation. Also, the growth of new roots was
poor. Ramson often forms dense populations covering
large areas in horn-beam-oak and beech forests, it can
even become monodominant over big areas in the herb
layer in the woods (Szafer and Zarzycki 1972; Cˇ incˇura
et al. 1990; Oborny et al. 2011; Morschhauser et al. 2009). A. ursinum is considered as one of the species,
the patchy distribution of which is a characteristic
feature of the herb layer in the Hordelymo-Fagetum
forest community. Under the dense carpets of these
species, competition for space and light is likely to
occur (Leuschner and Lendzion 2009). Furthermore,
A. ursinum is a strong inter-specific competitor, which
affects the growth of other herbaceous plants via soil,
where phenolic phytotoxins (produced by ramson) are
accumulated. It competes with other plant species also
through volatile compounds (Djurdjevic et al. 2004). Experimental data from the seeds germination and
seedlings growth tests have shown that aqueous
extract and volatile compounds of ramson bulbs
inhibited other plants (lettuce, amaranth and wheat)
growth stronger than extract from the leaves (Djurdj-
evic et al. 2004). with high assimilation rate, and accumulation of
nutrients stored in the bulbs (Jandl et al. 1997). The
above-ground parts abruptly wither as summer arrives. The mortality rate of A. ursinum during the first
2 years of life is estimated at around 21 % (Bierzych-
udek 1982). In the following year it increases as the
plant develops contractile roots which are more
subjected to attack by insects and nematodes (Ernst
1979). It is estimated that only about 1–10 % of
ramson seedlings reach the reproductive age (Bier-
zychudek 1982). The estimated average life span and
age of the first sexual reproduction for A. ursinum are
8–10 and 4–5 years respectively. Sulfur compounds These are undoubtedly the most important ramson’s
constituents, both in terms of chemotaxonomic value
and pharmacological activity. Their qualitative and
quantitative profile is subject to great variation, the
predominant reason for which is their lability. Of the
various sulfur compounds present in this, as well as
other Allium species, glutamyl peptides and sulfoxides
are considered as primary. Usually, these plants
contain a high concentration of S-alk(en)yl-L-cys-
teine-sulfoxides—odorless, non-volatile sulfur sec-
ondary metabolites, which, following subsequent
hydrolysis, give rise to many volatile compounds,
including thiosulphinates and (poly)sulfides responsi-
ble for specific Allium flavor and odor (Boscher et al. 1995; Yoo and Pike 1998). The period of active growth of A. ursinum lasts
3.5–4 months starting in early spring, between late
February and early March, before the full development
of tree leaves (e.g. in the northern Vienna Woods—
approximately 60 days before) (Jandl et al. 1997). This provides enough of light in the first stage of the
plant growth and helps to avoid competition for light
with the canopy (Shmanova and Krichfalushii 1995;
Oborny et al. 2011). In late spring however, the canopy
cover protects the plant from direct sunlight and helps
to keep the appropriate humidity of the topsoil air. Rapid development of ramson stands is associated Ramson belongs to a methiin/alliin-type Allium
species, which means it contains mainly a mixture of
(?)-S-methyl-L-cysteine-sulfoxide (methiin) and (?)-
S-allyl-L-cysteine-sulfoxide = (?)-S-2-propenyl-L-
cysteine-sulfoxide (alliin) (Schmitt et al. 2002; Kubec
et al. 2000). However, isoalliin—(?)-S-(1-propenyl)-
L-cysteine-sulfoxide
and
propiin—(?)-S-propyl-L-
cysteine-sulfoxide are present as well (Fig. 1) (Sch-
mitt et al. 2002). Also, ethiin (S-ethyl-cysteine-sulf-
oxide) was reported in the sample of fresh leaves 12 3 3 Phytochem Rev (2015) 14:81–97 84 collected in April in Czech Republic (Kubec et al. 2000)
CH3
S
COOH
O
NH2
Methiin
S
COOH
O
NH2
CH2
Alliin
CH3
S
COOH
O
NH2
Isoalliin
S
COOH
O
NH2
CH3
Propiin
S
COOH
O
NH2
CH3
Ethiin
Fig. 1 Allium ursinum
cysteine-sulfoxides showed that the highest amounts in leaves, storage
leaves and bulbs (0.42, 0.26, 0.38 % respectively)
were reached in March and April, that is before
flowering time (Schmitt et al. 2002, 2005). Samples of
fruits and leaf stalks collected in June contained 0.25
and 0.15 % of cysteine sulfoxides (fw) respectively. Furthermore, the relative quantitative profile of the
investigated sulfoxides (alliin, methiin, isoalliin and
propiin) differed during the vegetation period. Sulfur compounds In
March the bulb contained almost the same amounts of
alliin and methiin. In the following weeks alliin
became the main component (73 ± 18 %), while
methiin content decreased to 15 ± 9 % in mid-May
(Schmitt et al. 2005). Then, the rise of methiin levels
was observed. The relative content of propiin was
always below 5 %, while of isoalliin *10 %. Fig. 1 Allium ursinum
cysteine-sulfoxides Fig. 1 Allium ursinum
cysteine-sulfoxides As was mentioned above, cysteine sulfoxides are
subject to hydrolytic cleavage leading to a formation
of a number of characteristic volatile secondary
products. This is performed by specific enzymes,
named C,S-lyases, which catalyse the cleavage of the
Cb–Sc bond of the sulfoxides. In the intact cell,
cysteine sulfoxides are localized in the cytoplasm,
while the hydrolytic enzyme is found in vacuoles. Cellular compartmentalization damage results in its
release, and subsequent hydrolysis of sulfoxides
(Boscher et al. 1995; Sendl 1995). This reaction
occurs upon tissue damage, e.g. when the organ is
crushed, minced, or otherwise processed, or in case of
the pathogens attack (Ankri and Mirelman 1999). collected in April in Czech Republic (Kubec et al. 2000). collected in April in Czech Republic (Kubec et al. 2000). The quantitative profile of cysteine-sulfoxides
depends on the plant organ and time of harvest. Their
total content in the leaves collected in April, expressed
as mg/100 g of fresh weight, was 101.9 (in this:
methiin—60.0, ethiin—0.4, propiin—1.2, alliin—
40.3, isoalliin—traces) (Kubec et al. 2000). The
content of total cysteine sulfoxides in the bulbs
harvested in late summer, calculated as alliin, was
0.26 % (amount was related to the fresh weight)
(Keusgen et al. 2003). In water extracts from cloves
and leaves containing hydrolytic enzyme inhibitor, the
alliin content ranges were: for cloves—0.65–1.10 and
for leaves 0.20–0.72; while methiin: 0.60–1.40 and
0.30–0.95, respectively (Sendl 1995). The relative
proportions of cysteine sulfoxides in ramson are
presented in Table 1 (Fritsch and Keusgen 2006). C,S-lyase isolated from ramson has a molecular
mass of 150,000 Da and consists of three subunits
(Landshuter et al. 1994). The optimal pH and temper-
ature for its activity were established at 6.0 and 35 C,
respectively. After 30 min. of incubation at pH 3.0
ramson’s C,S-lyase lost its activity by 90 %. Never-
theless, its enzymatic properties are maintained at low
temperatures, what was observed even after ramson
cloves were freezed at -20 C. Sulfur compounds Table 2 Data from quantitative HPLC determination of thio-
sulfinates in chloroform extracts from leaves and bulbs of
Allium ursinum (adapted from Sendl and Wagner 1991)
Plant material
Allicin (%)
MATS (%)
DMTS (%)
A. ursinum bulbs
0.53
0.70
0.27
A. ursinum leaves
0.13
0.26
0.13
MATS allyl-methyl and/or methyl-allyl thiosulfinate, DMTS
dimethyl thiosulfinate Table 2 Data from quantitative HPLC determination of thio-
sulfinates in chloroform extracts from leaves and bulbs of
Allium ursinum (adapted from Sendl and Wagner 1991) Table 2 Data from quantitative HPLC determination of thio-
sulfinates in chloroform extracts from leaves and bulbs of
Allium ursinum (adapted from Sendl and Wagner 1991) Table 2 Data from quantitative HPLC determination of thio-
sulfinates in chloroform extracts from leaves and bulbs of
Allium ursinum (adapted from Sendl and Wagner 1991) Thiosulfinates are regarded as primarily responsi-
ble for odor and flavor of freshly prepared ramson
macerates, and of the products obtained by both the
extraction and an unorthodox kind of distillation
conducted at room temperature (Block et al. 1992). The results showed that room temperature steam
distillation provides half the amount of thiosulfinates
obtained by direct extraction (Block et al. 1992). The major thiosulfinates found in ramson extracts
are allicin (diallyl thiosulfinate = di-2-propenyl thio-
sulfinate), and methyl-allyl- or dimethyl thiosulfinates
(Fig. 2) (Sendl and Wagner 1991). According to Sendl
they constitute 75–90 % of all the compounds formed
immediately after hydrolysis of cysteine sulfoxides
(Sendl 1995). Data from quantitative HPLC determi-
nation of thiosulfinates in chloroform extracts from
leaves and bulbs, refered to dry weight, is presented in
Table 2 (Sendl and Wagner 1991). In a ramson freeze-
dried
powder
total
thiosulfinates
concentrations
expressed as molar percentage of total was 21 (based
on the weight of powder) (Block et al. 1992). are formed as reaction products in solvents less polar
than 2-propanol, e.g. hexane (Sendl 1995). It was also
observed that pure allicin due to its thermolabile
nature degraded to vinyldithiins during GLC analysis. In the hexane extract from the bulbs of A. ursinum
Wagner and Sendl (1990) identified 2-vinyl-4H-1,3-
dithiin and 3-vinyl-4H-1,2-dithiin (Fig. 3). Vinyldi-
thiins (3,4-dihydro-3-vinyl-1,2-dithiin and 2-vinyl-
4H-1,3-dithiin) were identified also in the essential
oil isolated from the leaves and flowers of ramson
samples collected in Bulgaria (Ivanova et al. 2009). Thiosulfinates are unstable, reactive compounds,
that easily decompose to (poly)sulfides, dithiins,
ajoenes, and other volatile and non volatile com-
pounds. Sulfur compounds It seems that such
feature of the C,S-lyase system provides all year long
protection for the underground parts of the plant after
disruption of the parenchyma. In contrast to similar The analysis of the changes in the total cysteine
sulfoxides content in different parts of A. ursinum
collected in Germany throughout the vegetation
period (focused on the months from March to June) Table 1 The relative
proportions of cysteine
sulfoxides in leaves and
bulbs of ramson (adapted
from Fritsch and Keusgen
2006)
Total content
(average) (%)
Relative
methiin (%)
Relative
alliin (%)
Relative
isoalliin (%)
Relative
propiin (%)
Origin of
material
0.245
39
24
37
0
Urft in
der Eifel
0.164
35
28
37
0
123 Total content
(average) (%)
Relative
methiin (%)
Relative
alliin (%)
Relative
isoalliin (%)
Relative
propiin (%)
Origin of
material
0.245
39
24
37
0
Urft in
der Eifel
0.164
35
28
37
0 12 3 Phytochem Rev (2015) 14:81–97 85 S S
O
CH2
CH2
Allicin
CH3
S S
O
CH2
Methyl-allyl thiosulfinate
S S CH3
O
CH2
Allyl-methyl thiosulfinate
CH3
S S CH3
O
Dimethyl thiosulfinate
Fig. 2 Major thiosulfinates
found in ramson extracts
Table 2 Data from quantitative HPLC determination of thio-
sulfinates in chloroform extracts from leaves and bulbs of
Allium ursinum (adapted from Sendl and Wagner 1991)
Plant material
Allicin (%)
MATS (%)
DMTS (%)
A. ursinum bulbs
0.53
0.70
0.27
A. ursinum leaves
0.13
0.26
0.13
MATS allyl-methyl and/or methyl-allyl thiosulfinate, DMTS
dimethyl thiosulfinate Fig. 2 Major thiosulfinates
found in ramson extracts Fig. 2 Major thiosulfinates
found in ramson extracts enzymes isolated from A. sativum, A. cepa or A. porrum, ramson’s C,S-lyase is not a glycoprotein. This
was revealed as no interaction with Concanavalin-A-
Sepharose, and no staining by periodic acid Schiff-
reagent, were observed. The enzyme was however
sensitive to low hydroxylamine concentrations. The
ramson’s C,S-lyase catalyses S-alk(en)yl-cysteine-
sulfoxides hydrolysis, with alliin as the most preferred
substrate (Landshuter et al. 1994). Its comparison with
common garlic’s alliinase showed that even though it
is less specific to alliin, it still shows higher relative
activity towards other cysteine sulfoxides, especially
methiin and isoalliin (Schmitt et al. 2005). The
primary products formed as a result of C,S-lyases
action are thiosulfinates, pyruvic acid, and ammonia. Fig. 2 Major thiosulfinates
found in ramson extracts Sulfur compounds This takes place on storage, during process-
ing, e.g. in the presence of organic solvents, and also
when heat-treated. Allicin is very unstable even at
room temperature. Studies by Bagiu et al. (2010)
showed that after 20 h at 20 C it decomposed
completely resulting in di-2-propenyl disulfide, di-2-
propenyl trisulfide, di-2-propenyl sulfide, and sulfur
dioxide. Another group of thiosulfinates degradation pro-
ducts, namely ajoene, methyl- and dimethyl ajoenes,
were identified in acetone–chloroform extracts from
ramson bulbs (Fig. 3). Comparative analysis of A. sativum and A. ursinum extracts showed that ajoene
dominated in garlic, while its methyl- and dimethyl
homologues were the main components in ramson
extract (Wagner and Sendl 1990). It should be
mentioned that ajoenes and vinyldithiins were found
as well in oil-macerated garlic (Benkeblia and Lan-
zotti 2007). Vinyldithiins are cyclic compounds which are
another group of degradation products of allicin. They Apart from the above-mentioned sulfur-containing
sulfoxides
degradation
products,
various
sulfur 12 3 Phytochem Rev (2015) 14:81–97 86 Table 3 Composition of essential oils isolated from Allium
ursinum leaves collected at different sites in Europe (Ivanova
et al. 2009; Godevac et al. 2008; Schmitt et al. 2005;
Bła_zewicz-Woz´niak and Michowska 2011)
Compounds
Origin of plant material
*1
*2
*3
*4
*5
*6
Sulfide, methyl 2-propenyl
? Sulfide, di-2-propenyl
? Propylene sulfide
? Disulfide, methyl 2-propenyl
? ? ? ? ? ? Disulfide, methyl propyl
? ? ? ? ? Disulfide, methyl 1-propenyl,
(E)-
? ? ? ? ? Disulfide, methyl 1-propenyl,
(Z)-
? Disulfide, dimethyl
? ? ? ? ? Disulfide, dipropyl
? Hex-3-en-1-ol, (E)-
? ? Disulfide, di-2-propenyl
? ? ? ? ? ? Disulfide, 2-propenyl propyl
? ? ? Allyl (E)-1-propenyl disulfide
? ? Disulfide, 1-propenyl propyl, cis
? Disulfide, 1-propenyl propyl,
trans
? Nonanal
? ? ? Trisulfide, methyl 2-propenyl
? ? ? ? ? Trisulfide, methyl propyl
? ? Trisulfide, dimethyl
? ? ? ? ? ? Trisulfide, methyl 1-propenyl,
(Z)-
? Trisulfide, methyl 1-propenyl,
(E)-
? Tetrasulfide, dimethyl
? ? ? ? ? Trisulfide, dipropyl
? ? Trisulfide, di-2-propenyl
? ? ? ? ? Trisulfide, propyl 2-propenyl
? Trisulfide, 1-propenyl
2-propenyl, (Z)-
? Trisulfide, 1-propenyl
2-propenyl, (E)-
? Tetrasulfide, methyl 2-propenyl
? ? ? ? Tetrasulfide,di-2-propenyl
? ? ? ? ? 1-Propene, 3,30-thiobis-
(CAS#592-88-1)
? ? Propylthiol
? Dimethylthiophene
? ? Pentanal, 2-methyl
? 2-Hexenal
? ? Cumene
? ? Sulfur compounds R S
S S R'
O
R, R' = allyl; Ajoene
R, R' = allyl, methyl; Methyl ajoene
R, R' = methyl; Dimethyl ajoene
S
S
CH2
2-Vinyl-4H-1,3-dithiin
S
S
CH2
3-Vinyl-4H-1,2-dithiin
Fig. 3 Ajoenes and dithiins present in A. ursinum extracts R S
S S R'
O
R, R' = allyl; Ajoene
R, R' = allyl, methyl; Methyl ajoene
R, R' = methyl; Dimethyl ajoene Table 3 Composition of essential oils isolated from Allium
ursinum leaves collected at different sites in Europe (Ivanova
et al. 2009; Godevac et al. 2008; Schmitt et al. 2005;
Bła_zewicz-Woz´niak and Michowska 2011) Table 3 Composition of essential oils isolated from Allium
ursinum leaves collected at different sites in Europe (Ivanova
et al. 2009; Godevac et al. 2008; Schmitt et al. 2005;
Bła_zewicz-Woz´niak and Michowska 2011) 2-Vinyl-4H-1,3-dithiin S
S
CH2
3-Vinyl-4H-1,2-dithiin Fig. 3 Ajoenes and dithiins present in A. ursinum extracts compounds have also been detected as constituents of
ramson’s essential oil. This was obtained (0.007 %)
for the first time already in 1887 by Semmler, who
identified alkyl sulfides, alkyl polysulfides, and trace
amounts of mercaptan (Sendl 1995). The amount of oil varies depending on soil
condition, geographical location, and part used. For
example, isolation of essential oil from fresh and air-
dried leaves and flowers of ramson collected in Bosnia
yielded 0.011 % (w/w) and 0.024 % (w/w) respec-
tively (Copra-Janicijevic et al. 2008). The results of
qualitative analyses of essential oils from ramson
collected from different sites in Europe showed
significant differences in their composition (Table 3). Furthermore, different time of harvest and analytical
method applied influenced the profile of investigated
oils. From among over 20 components identified in the
volatile oil of A. ursinum collected in Serbia, the most
abundant fraction was disulfides (54.7 %), followed by
trisulfides (37.0 %), tetrasulfides (4.7 %), and the non-
sulfur components (1.0 %) (Godevac et al. 2008). The
composition of essential oils of three ecotypes of A. ursinum collected in Poland also differed significantly
in the dominant components (Bła_zewicz-Woz´niak and
Michowska 2011). The Roztocze ecotype contained
methyl-2-propenyl disulfide (16.05 % on average),
6,10,14-trimethyl-2-pentadecanone (13.55 %), non-
anal (11.93 %), and dimethyl trisulfide (12.07 %) as
main components. Sulfur compounds Table 3 continued
Compounds
Origin of plant material
*1
*2
*3
*4
*5
*
Pentacosane
? Hexacosane
? Heptacosane
? Nonacosane
? Phytol isomer
? tr. Traces
*1 Leaves harvested in the vicinity of Ihtiman, Bulgaria; the o
obtained
by
hydrodistillation
method;
GC/MS
analysi
(Ivanova et al. 2009)
*2 Leaves collected in the vicinity of Belgrade, Serbia
hydrodistillation; GC/MS analysis (Godevac et al. 2008)
*3 The samples collected in the area of Quedlinburg, Germany
the investigation by SPME-GC (Schmitt et al. 2005)
*4 The plant material harvested in Dukla, Poland
*5 Roztocze region, Poland
*6
Bieszczady,
Poland;
isolation
of
the
oil
by
steam
distillation;
GC/MS
analysis
(Bła_zewicz-Woz´niak
an
Michowska 2011) Table 3 continued
Compounds
Origin of plant material
*1
*2
*3
*4
*5
*6
Benzene, 1-ethyl-4-methyl
? 1,2,3-Trimethylbenzene
tr. ? ? 5-Hepten-2-one, 6-methyl
? n-Octanal
tr. ? ? Cyclohexanone, 2,2,6-trimethyl
? ? 1-Propene, 1-(methylthio)-, (Z)-
? ? ? 1-Propene, 1-(methylthio)-, (E)-
? ? tr. Decenal
? ? tr. 1-Cyclohexene-1-acetaldehyde,
2,2,6-trimethyl
tr. ? 2H-1-benzopyran 3,4,4a,5,6,8a-
hexahydro-2,5,5,8a-
tetramethyl, (2a,4a a a,8aa)-
? ? ? 2,6,10,10-Tetramethyl-1-oxa-
spiro[4,5]dec-6-ene
? ? 2-Undecanone, 6,10-dimethyl
tr. ? (E)-b-caryophyllene
? ? Geranyl acetone
? ? ? Alloaromadendrene
? ? (E)-b-ionone
? ? ? 2-Tridecanone
? ? ? 1,6,10-Dodecatrien-3-ol, 3,7,11-
trimethyl
? 2-Hexadecanol
? Spathulenol
? ? ? Caryophyllene oxide
? ? ? 1-Tetradecanal
? ? ? ? Pentadecanal
? ? ? 2-Pentadecanone, 6,10,14-
trimethyl
? ? ? 5,9,13-Pentadecatrien-2-one,
6,10,14-trimethyl, (E,E)-
? ? ? Phytol
? ? ? ? n-Hexadecanoic acid
? ? ? Phytol acetate
? ? ? 2-Vinyl-1,3-dithiane
? 3,4-Dihydro-3-vinyl-1,2-dithiin
? 2-Vinyl-4H-1,3-dithiin
? 2-Hexenol
? 1-Hexadecanal
? 1-Octadecenol
? 1-Octadecen
? Heneicosane
? Table 3 continued
Compounds
Origin of plant material
*1
*2
*3
*4
*5
*6
Pentacosane
? Hexacosane
? Heptacosane
? Nonacosane
? Phytol isomer
? tr. Traces
*1 Leaves harvested in the vicinity of Ihtiman, Bulgaria; the oil
obtained
by
hydrodistillation
method;
GC/MS
analysis
(Ivanova et al. 2009)
*2 Leaves collected in the vicinity of Belgrade, Serbia;
hydrodistillation; GC/MS analysis (Godevac et al. 2008)
*3 The samples collected in the area of Quedlinburg, Germany;
the investigation by SPME-GC (Schmitt et al. Sulfur compounds The Dukla ecotype oil was com-
posed mainly of phytol (17.03 %) and n-hexadecanoic
acid (16.57 %), while in the oil of the Bieszczady
ecotype phytol acetate (16.40 %) and (E)-b-ionone
(13.33 %)
dominated
(Bła_zewicz-Woz´niak
and 12 123 Phytochem Rev (2015) 14:81–97 87 Michowska 2011). The results of SPME-GC analysi
of ramson oil, from the leaves collected in the area o
Quedlinburg (Germany) showed that diallyl disulfid
was the major component, amounting to *50 %
(Schmitt et al. 2005). Allyl-methyl disulfide, allyl
methyl sulfide, diallyl sulfide, and (E)-allyl-1-propeny
disulfide were abundant, as well. However, accordin
to the authors, the SPME method did not supply
information on some other volatile substances, such a
2-hexenal or hex-3-en-1-ol, which were detected by an
SDE-GC method. GC/MS analysis of samples from
fresh flowers collected in the vicinity of Ihtiman
(Bulgaria), showed that the main components of th
volatile fraction were (E)-methyl-2-propenyl disulfide
methyl-2-propenyl trisulfide, dimethyl trisulfide, 3,4
dihydro-3-vinyl-1,2-dithiin and 2-vinyl-4H-1,3-dith
iin (Ivanova et al. 2009). Schmitt et al. (2005) detected
the decrease of the relative yield of oil (sum of volatil
substances) during the vegetation period, and observed
that this was in agreement with the decreasing amount
of cysteine sulfoxides. An especially significan
decrease in allyl methyl disulfide and dimethyl disul
fide levels was seen, while the relative content of (E)
allyl-1-propenyl disulfide increased. Apart from studies on the composition of steam
distilled essential oil from ramson, a very interesting
Table 3 continued
Compounds
Origin of plant material
*1
*2
*3
*4
*5
*6
Benzene, 1-ethyl-4-methyl
? 1,2,3-Trimethylbenzene
tr. ? ? 5-Hepten-2-one, 6-methyl
? n-Octanal
tr. ? ? Cyclohexanone, 2,2,6-trimethyl
? ? 1-Propene, 1-(methylthio)-, (Z)-
? ? ? 1-Propene, 1-(methylthio)-, (E)-
? ? tr. Decenal
? ? tr. 1-Cyclohexene-1-acetaldehyde,
2,2,6-trimethyl
tr. ? 2H-1-benzopyran 3,4,4a,5,6,8a-
hexahydro-2,5,5,8a-
tetramethyl, (2a,4a a a,8aa)-
? ? ? 2,6,10,10-Tetramethyl-1-oxa-
spiro[4,5]dec-6-ene
? ? 2-Undecanone, 6,10-dimethyl
tr. ? (E)-b-caryophyllene
? ? Geranyl acetone
? ? ? Alloaromadendrene
? ? (E)-b-ionone
? ? ? 2-Tridecanone
? ? ? 1,6,10-Dodecatrien-3-ol, 3,7,11-
trimethyl
? 2-Hexadecanol
? Spathulenol
? ? ? Caryophyllene oxide
? ? ? 1-Tetradecanal
? ? ? ? Pentadecanal
? ? ? 2-Pentadecanone, 6,10,14-
trimethyl
? ? ? 5,9,13-Pentadecatrien-2-one,
6,10,14-trimethyl, (E,E)-
? ? ? Phytol
? ? ? ? n-Hexadecanoic acid
? ? ? Phytol acetate
? ? ? 2-Vinyl-1,3-dithiane
? 3,4-Dihydro-3-vinyl-1,2-dithiin
? 2-Vinyl-4H-1,3-dithiin
? 2-Hexenol
? 1-Hexadecanal
? 1-Octadecenol
? 1-Octadecen
? Heneicosane
? Tricosane
? Tetracosane
? Phenolics As far as qualitative profile is concerned, ramson is
abundant predominantly in kaempferol derivatives. The ethanol extract from the leaves collected near
Laceno Lake (Italy) yielded: 3-O-b-neohesperidoside-
7-O-[2-O-(trans-p-coumaroyl)]-b-D-glucopyranoside
(1), 3-O-b-neohesperidoside-7-O-[2-O-(trans-p-feru-
loyl)]-b-D-glucopyranoside (2), 3-O-b-neohesperido-
side-7-O-[2-O-(trans-p-coumaroyl)-b-D-glucopyran-
osyl]-b-D-glucopyranoside (3), 3-O-b-neohesperido
side-7-O-b-D-glucopyranoside (4), 3-O-b-neohesperi-
doside (5) (Carotenuto et al. 1996). From the n-butanol
fraction of the dry leaves of ramson collected in
Denmark seven flavonoid glycosides were isolated. Three of them have been previously reported by
Carotenuto et al. (compounds 1, 4, 5). The remaining
were also identified as kaempferol derivatives: 3-O-b-
D-glucopyranoside,
3-O-b-D-glucopyranosyl-7-O-b-
D-glucopyranoside, 3-O-a-L-rhamnopyranosyl-(1 ? 2)-
[3-acetyl]-b-D-glucopyranoside and 3-O-a-L-rhamno-
pyranosyl-(1 ? 2)-[6-acetyl]-b-D-glucopyranoside
(Wu et al. 2009). Compounds 4, 5, kaempferol 3-O-b-
D-glucopyranoside and kaempferol 3-O-b-D-glucopyr-
anosyl-7-O-b-D-glucopyranoside were isolated from
the n-butanol extract from fresh flowers (Ivanova et al. 2009). The analysis of flavonoid content in acidified
methanol extracts from green and yellow leaves, stalks
and seeds collected in June in the forest area near
Wrocław (Poland) led to the isolation of 21 com-
pounds, all kaempferol derivatives (Oszmian´ski et al. 2013). Apart from sulfur-containing substances A. ursinum
has been also reported to be a good source of phenolic
compounds. It should be mentioned, however that the
extraction method may substantially alter the level of
active compounds isolated. Total polyphenol content,
expressed as gallic acid equivalents (GAE), in the leaf
extract obtained by a 12-day maceration with 70 %
ethanol at room temperature (20 C) was higher in
comparison with the one prepared by the ultrasound-
assisted extraction: 27.9 g GAE/100 g dry basis
versus *10 g GAE/100 g (Gıˆtin et al. 2012). Total
free phenolics content in the leaves was determined as
3.24 mg/g, while in the bulbs 2.30 mg/g. The amount
of bound forms was about the same in the leaves and in
the bulbs (1.10 and 1.00 respectively) (Djurdjevic
et al. 2004). Gross differences were also noted in
studies on gallic acid levels. Its qualitative and
quantitative analysis in hydroalcoholic extracts from
A. ursinum leaves showed that: 96 % methanol extract
had a gallic acid content 0.0576 mg/ml, 80 % meth-
anol extract—0.0165 mg/ml; while 96 % ethanol
extract—0.0076 mg/ml (Condrat et al. 2010). The studies on the content of phenolic acids in fresh
leaves and bulbs of ramson collected in an experi-
mental forest situated in West Serbia exhibited some
differences between free and bound compounds in
these plant parts (Djurdjevic et al. 2004). Sulfur compounds Such studies were
conducted in a Viennese suburban forested park in
which A. ursinum was grown as ground cover (Pux-
baum and Ko¨nig 1997). Sulfur emission rates (lg S)
per gram of dry weight and per unit of ground area were
1 lg/g 9 h and 60 lg/m2 9 h, respectively. The
authors claimed this was the highest rate ever reported
for such substances emitted from a terrestrial plant. Phenolics The amounts
of free phenolic acids in leaves and bulbs were 119.75
and
180.91 lg/g,
respectively,
while
of
bound
forms—135.30 and 248.97 lg/g, respectively. The
leaves contained free forms of ferulic and vanillic
acids, and bound forms of p-coumaric, ferulic and
vanillic acids. In the bulbs free ferulic, p-hydroxy-
benzoic and vanillic acids, and bound forms of p-
coumaric and ferulic acids were detected. Sulfur compounds 2005)
*4 The plant material harvested in Dukla, Poland
*5 Roztocze region, Poland
*6
Bieszczady,
Poland;
isolation
of
the
oil
by
steam
distillation;
GC/MS
analysis
(Bła_zewicz-Woz´niak
and
Michowska 2011) *
Benzene, 1-ethyl-4-methyl
1,2,3-Trimethylbenzene
5-Hepten-2-one, 6-methyl
n-Octanal
Cyclohexanone, 2,2,6-trimethyl
1-Propene, 1-(methylthio)-, (Z)-
1-Propene, 1-(methylthio)-, (E)-
Decenal
1-Cyclohexene-1-acetaldehyde,
2,2,6-trimethyl
2H-1-benzopyran 3,4,4a,5,6,8a-
hexahydro-2,5,5,8a-
tetramethyl, (2a,4a a a,8aa)-
2,6,10,10-Tetramethyl-1-oxa-
spiro[4,5]dec-6-ene
2-Undecanone, 6,10-dimethyl
(E)-b-caryophyllene
Geranyl acetone
Alloaromadendrene
(E)-b-ionone
2-Tridecanone
1,6,10-Dodecatrien-3-ol, 3,7,11-
trimethyl
2-Hexadecanol
Spathulenol
Caryophyllene oxide
1-Tetradecanal
? Pentadecanal
2-Pentadecanone, 6,10,14-
trimethyl
5,9,13-Pentadecatrien-2-one,
6,10,14-trimethyl, (E,E)-
Phytol
? n-Hexadecanoic acid
Phytol acetate
2-Vinyl-1,3-dithiane
? 3,4-Dihydro-3-vinyl-1,2-dithiin
? 2-Vinyl-4H-1,3-dithiin
? 2-Hexenol
? 1-Hexadecanal
? 1-Octadecenol
? 1-Octadecen
? Heneicosane
? Tricosane
? Tetracosane
? Michowska 2011). The results of SPME-GC analysis
of ramson oil, from the leaves collected in the area of
Quedlinburg (Germany) showed that diallyl disulfide
was the major component, amounting to *50 %
(Schmitt et al. 2005). Allyl-methyl disulfide, allyl-
methyl sulfide, diallyl sulfide, and (E)-allyl-1-propenyl
disulfide were abundant, as well. However, according
to the authors, the SPME method did not supply
information on some other volatile substances, such as
2-hexenal or hex-3-en-1-ol, which were detected by an
SDE-GC method. GC/MS analysis of samples from
fresh flowers collected in the vicinity of Ihtiman
(Bulgaria), showed that the main components of the
volatile fraction were (E)-methyl-2-propenyl disulfide,
methyl-2-propenyl trisulfide, dimethyl trisulfide, 3,4-
dihydro-3-vinyl-1,2-dithiin and 2-vinyl-4H-1,3-dith-
iin (Ivanova et al. 2009). Schmitt et al. (2005) detected
the decrease of the relative yield of oil (sum of volatile
substances) during the vegetation period, and observed
that this was in agreement with the decreasing amounts
of cysteine sulfoxides. An especially significant
decrease in allyl methyl disulfide and dimethyl disul-
fide levels was seen, while the relative content of (E)-
allyl-1-propenyl disulfide increased. (E)-b-ionone Apart from studies on the composition of steam-
distilled essential oil from ramson, a very interesting 12 3 3 Phytochem Rev (2015) 14:81–97 88 in March, from the Bacau city forests (Romania), using
ultrasound-assisted extraction was *7.3 mg QE/kg
fresh plant; while using conventional maceration—
2.7 mg QE/kg (Gıˆtin et al. 2012). Also, the total
content of flavonoids in the different parts of A. ursinum collected in June in the forest area near
Wrocław (Poland) differed significantly: seeds—
73.14 mg/100 g of dry mass, stalks—206.07 mg/
100 g,
green
leaves—1,856.31 mg/100 g,
yellow
leaves 2,362.96 mg/100 g (Oszmian´ski et al. 2013). aspect is investigation of the atmospheric emission rate
of organic sulfur compounds. Steroidal glycosides Similarly to organosulfur compounds, steroidal saponins
are also commonly found in the Allium genus. The
following were reported in the bulbs of A. ursinum:
diosgenin 3-O-a-L-rhamnopyranosyl-(1 ? 4)-a-L-rhamno
pyranosyl-(1 ? 4)-[a-L-rhamnopyranosyl-(1 ? 2)]-
b-D-glucopyranoside and (25R)-spirost-5,25(27)-dien- The flavonoid content (expressed as mg quercetin
equivalent—QE) determined in fresh leaves collected 123 123 123 89 Phytochem Rev (2015) 14:81–97 O
O
OH
O
H
O
H
Me
O
O
O
H
O
OH
O
OH
O
H
O
Me
O
OH
O
H
O
H
Me
O
O
R Fig. 4 Steroidal glycosides
isolated from the bulbs of A. ursinum Fig. 4 Steroidal glycosides
isolated from the bulbs of A. ursinum R = CH3; Diosgenin 3-O-α-L-rhamnopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→4)-
α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside
[ R = CH3; Diosgenin 3-O-α-L-rhamnopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→4)-
α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside
[ R = CH2; (25R)-spirost-5,25(27)-dien-3β-ol 3-O-α-L-rhamnopyranosyl-(1→4)-α-L-
rhamnopyranosyl-(1→4)-[α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside O
O
O
O
OH
O
H
O
H
Me
O
O
H
O
OH
O
OH
O
H
O
Me
O
OH
O
H
O
H
Me
3-Hydroxy-pregna-5,16-dien-20-on 3-O-α-L-rhamnopyranosyl-(1→4)-α-L-
rhamnopyranosyl-(1→4)-[α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside 3-Hydroxy-pregna-5,16-dien-20-on 3-O-α-L-rhamnopyranosyl-(1→4)-α-L-
rhamnopyranosyl-(1→4)-[α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside and leaves collected in April (Smeets et al. 1997). Root
compounds were identical to those found in bulbs:
AUAI, which is a heterodimeric lectin composed of
polypeptides of 12.5 and 11.5 kDa, and AUAII a
homodimeric lectin composed of polypeptides of
12 kDa. Both lectins are mannose-specific, and show
a good reaction with synthetic (1 ? 3) and (1 ? 6)
mannans. The ramson leaf lectin (AUAL) differs from
the bulb lectins, and also from the leaf-specific lectins
identified in other Allium species. It is a dimer
composed of 12 kDa subunits. 3b-ol 3-O-a-L-rhamnopyranosyl-(1 ? 4)-a-L-rhamno-
pyranosyl-(1 ? 4)-[a-L-rhamnopyranosyl-(1 ? 2)]-b-
D-glucopyranoside (Fig. 4) (Sobolewska et al. 2006). A
pregnane glycoside: 3-hydroxy-pregna-5,16-dien-20-on
3-O-a-L-rhamnopyranosyl-(1 ? 4)-a-L-rhamnopyrano-
syl-(1 ? 4)-[a-L-rhamnopyranosyl-(1 ? 2)]-b-D-gluco-
pyranoside has been identified as well (Fig. 4)
(Sobolewska et al. 2006). Diosgenin content in A. ursinum depended on the part
of the plant and the time of harvest (Sobolewska et al. 2009). Methanol extract prepared from fresh bulbs
collected in April, prior to flowering, yielded the highest
content of diosgenin (0,137 %). In the extract made from
leaves collected at the same time, the amount of diosgenin
was10timeslower,whileinthe one fromleavescollected
in March and June it was not detectable. Low diosgenin
content in ramson does not make this plant species a
valuable source for the isolation of this sapogenin. Thebulbsarealsorichinpolysaccharides.According
to Hegnauer, they may contain as much as 30–90 % of
mostly fructans (Hegnauer 1963). Fig. 4 Steroidal glycosides
isolated from the bulbs of A.
ursinum Steroidal glycosides In the bulbs harvested
between May and August the content of fructan U,
consisting of fructose residues only, was estimated as
75–90 % (Meier and Reid 1982). Unfortunately, no
modern structure elucidation studies on this compound
have been performed to date. The studies on reserve
carbohydrates in A. ursinum from the Sheffield flora
(United Kingdom) resulted in determination of the
maximum fructan concentration in the bulbs harvested
in summer, as 139 mg/g fresh weight (Hendry 1987). In an ethanol extract from fresh leaves b-sitosterol
3-O-b-D-glucopyranoside was found (Sabha et al. 2012). Uses Uses Ramson has been used for centuries to promote
general health, and as the old English proverb says: Eat leeks in Lide [March] and ramsons in May
And all the year after the physicians may play. Fig. 5 2-Di-O-a-linolenoyl-3-O-b-D-galactopyranosyl-sn-gly-
cerol (DLGG) found in an ethanol extract from fresh leaves of A. ursinum There is good evidence for the use of ramson by
Mesolithic people. Charred bulbs of A. ursinum were
identified—in the late Mesolithic settlement at Halss-
kov in Denmark (Kubiak-Martens 2002). It was
hypothesized that ramson was one of the plants that
contributed to the hunter-gatherer diet. A. ursinum was
known to the early Celts and to the ancient Romans. The Greek physician Dioscorides mentioned four
kinds of onion, among them A. ursinum and also
attributed a detoxifying effect to the plant (Meyer et al. 1999; Richter 1999). Ramson was well known also in
the Middle Ages; it belongs to the group of plants often
found at medieval West Slavic archeological sites
(Celka 2011). King Charles the Great, also known as
Charlemagne, included A. ursinum in his Capitulare
de Villis imperialibis, where he formally cataloged
plants, mostly those possessing medicinal properties,
and documented how the gardens should be planned
and cared for (Clickner 2011). Hieronymus Bock
provided drawings of the plant in his Kreutterbuch,
Lonicerus judged wild garlic to be superior to regular
garlic (Richter 1999; Bła_zewicz-Woz´niak et al. 2011;
Strzelecka and Kowalski 2000; Madaus 1938). oleic, palmitoleic, stearic, a-linolenic, and myristic
acid (Wiater et al. 1998). Moreover, water extracts
yielded fairly rare, but pharmacologically valuable c-
glutamylpeptides, and many amino acids, such as:
asparagine, glutamine, aspartic acid, glutamic acid,
arginine, alanine, glycine, threonine (Wagner and
Sendl 1990). In an ethanol extract from fresh leaves
2-di-O-a-linolenoyl-3-O-b-D-galactopyranosyl-sn-
glycerol (DLGG) (Fig. 5), was identified (Sabha et al. 2012). Ramson leaves seem to be relatively abundant in
pigments, as compared to other Allium plants, the
content of which amounts to: 2.87 ± 0.03 mg/g of
chlorophyll a, 1.35 ± 0.01 mg/g of chlorophyll b,
and as much as 9.99 ± 0.01 mg/g of carotenoids
(Sˇtajner and Szo¨llosi Varga 2003). Comparative
analysis of some macro- and microelements in A. ursinum and A. sativum showed that ramson con-
tained higher levels of magnesium (7,000 mg/kg),
manganese
(1,600 mg/kg)
and
iron
(230/mg/kg)
than garlic (6,114, 952, 14 mg/kg, respectively). A. ursinum is a rich source of adenosine (120 mg/kg)
(Nagori et al. 2010). Other Other interesting constituents identified in A. ursinum
include lectins, which were isolated from bulbs, roots A number of fatty acids were reported in the hexane
extract from the bulbs. These were palmitic, linoleic, 12 3 3 Phytochem Rev (2015) 14:81–97 90 O
OH
O
H
O
O
H
OH
CH2
C
H
H2C
O
O
O
CH3
O
CH3
Fig. 5 2-Di-O-a-linolenoyl-3-O-b-D-galactopyranosyl-sn-gly-
cerol (DLGG) found in an ethanol extract from fresh leaves of A. ursinum Uses Uses All parts of the plant are edible. For medical
purposes leaves/herb—Allii ursini folium/herba, col-
lected in April and May, and bulbs—Allii ursini
bulbus, collected in September and October, are used. Ramson is usually collected from the wild. However,
in Poland this species, which belongs to the group of
11 alliaceous plants growing wild there, has been
partially protected since 2004 and is listed in the ‘‘Red
list of plants and fungi in Poland’’, what made it
impossible to be wild-harvested (Szafer et al. 1988;
Zarzycki and Mirek 2006). Allium ursinum L. ssp. ucrainicum floral nectar
volume and concentration were investigated in three
different habitats in the Mecsek hills (South Trans-
danubia, Hungary) (Farkas et al. 2012). The study
revealed that ramson produces low to medium
volumes (ranged 0.1–3.8 ll) of highly concentrated
nectar (25–50 % sugar concentration). Freely sun-
exposed flowers produced lower quantity of nectar
than covered flowers at a given time. The higher
volume of nectar with higher sugar content was
observed in populations living in optimal life condi-
tions for A. ursinum (the sessile oak-hornbeam
association). The plants living in the silver lime-
flowering ash rock forest, where the lack of sufficient
nutrients was observed, produced lower quantities of
nectar. In European traditional medicine ramson has been
generally recommended as digestive stimulant, anti-
microbial agent, removing toxins from the body, and
to prevent cardiovascular diseases (Treben 1992;
Macku˚ and Krejcˇa 1989; Leporatti and Ivancheva
2003). It was often applied as a remedy in respiratory
problems, such as common cold with fever or
bronchitis. A. ursinum has been effective when used 12 3 Phytochem Rev (2015) 14:81–97 91 O
OH
O
OH
OH
O
OH
O
RO
OH
O
O
OH
O
H
OH
CH3
R = H; Kaempferol 3-O-β-neohesperidoside
R = Glc; Kaempferol 3-O-β-neohesperidoside-7-O-β-D-glucopyranoside O
OH
O
OH
OH
O
OH
O
RO
OH
O
O
OH
O
H
OH
CH3
β externally to support wound healing, in chronic skin
disorders, and in acne. In recent years there has been a growing interest in
its use as a dietary supplement and food. There are
some records that in the nineteenth century Switzer-
land butter made from milk of cows fed on ramson
were used. Such milk tasted slightly of garlic. Apparently in Eberbach in Germany there is a festival
called Ba¨rlauchtage—Bear’s Garlic Days, which is
devoted to this plant. Uses Today, it is a common practice to
use ramson in cuisine. Fresh leaves can be eaten raw or
cooked, and as a kind of pesto. They are often added to
soups, gnocchi, risotto, ravioli, and as a spice to flavor
hard cheeses or spreads based on cottage cheeses. Leaves and flowers can be used as a garnish to salads,
while ramson’s bulbs can be used like common garlic. R = H; Kaempferol 3-O-β-neohesperidoside R = H; Kaempferol 3-O-β-neohesperidoside R = Glc; Kaempferol 3-O-β-neohesperidoside-7-O-β-D-glucopyranoside Fig. 6 Flavonoids exerting in vitro inhibitory activity on
platelet aggregation induced by collagen Moreover, two of the flavonoids present in ramson
leaves: kaempferol 3-O-b-neohesperidoside-7-O-b-D-
glucopyranoside
and
3-O-b-neohesperidoside
(Fig. 6), showed in vitro inhibitory activity on platelet
aggregation induced by collagen (Carotenuto et al. 1996). As other kaempferol glycosides were inactive,
it was concluded that the presence of p-coumaroyl
group in the molecule and the increase in the number
of monosaccharides in the sugar residue deplete the
antiplatelet potential of these compounds. Allium ursinum is also a component of dietary
supplements available on the European market. For
example, it is one of constituents found in the recipes
used therapeutically in the University Hospital of
Bucharest (Romania) (Epure et al. 2011). Such
products as Api Ursomax and Memo Ursomax are
recommended as detoxifying and antiatherogenic
medicines. The former is additionally advertised as a
mineralizing agent, while Memo Ursomax is claimed
to be a memory stimulant. Ramson’s administration affects also the activity of
ACE. In vitro tests on the water extract from the leaves
(at the concentration of 0.3 mg/ml), showed higher
inhibition of this enzyme activity as compared to
garlic leaves extract (58 vs. 30 %) (Sendl et al. 1992a). This probably resulted from the differences in glut-
amyl peptides contents. The in vitro study on the effect
of the ramson essential oil on the artificial liposome
membrane model demonstrated that the fluidity of the
membrane close to the surface was statistically non-
significantly changed, while in deeper layers the
fluidity increased (Godevac et al. 2008). The authors
postulated that further studies should be continued to
estimate the role of A. ursinum volatile oil in the
regulation of membrane functions in hypertension. Pharmacological studies sativum
extracts (Sendl et al. 1992a). ramson was more effective than with garlic at the same
concentration (the final SBP—175 ± 1.2 mm Hg). A. ursinum decreased elevated circulating insulin con-
centration and total cholesterol level, however HDL
tended to increase. Similarly, when both garlics were
consumed at lower concentrations—0.1 % (w/w)—
systolic blood pressure readings at 10, 18, and 26 days
were significantly lower in rats consuming ramson
compared to the animals consuming common garlic. Authors concluded that these effects may be associ-
ated with high concentration of glutamyl peptides,
adenosine or phenolic compounds in ramson. They
suggested that consuming A. ursinum may result in a
greater therapeutic benefit when compared to A. sativum at a given concentration. Animal studies
demonstrated that ramson-containing diet may reduce
the size of the ischemic zone and ischemia and
reperfusion—induced arrhythmias (Rietz et al. 1993). microbes. It inhibited the growth of the following
bacteria: Staphylococcus aureus, Bacillus subtilis,
Escherichia coli, Proteus mirabilis, Salmonella ente-
ritidis, and fungi: Cladosporium sp., Aspergillus
niger, Rhizopus nigricans, Geotrichum candidum,
Penicillium expansum, Candida lipolytica, Mycoder-
ma, Saccharomycopsis fibuligera (Synowiec et al. 2010). The average antibacterial MIC value was
35 mg/ml with the exception of S. aureus ATTC
25923 strain, in the case of which the MIC was
17.7 mg/ml. The
highest
antifungal
effect
was
observed against C. lipolytica (MIC = 8.9 mg/ml),
whereas for other tested strains it was less pronounced
(MIC = 17.7 mg/ml), however still much higher in com-
parison to the water extract (concs. 41.9–83.7 mg/ml). The antibacterial activity of the water extract was seen
only against B. subtilis ATTC 6633 (at 83.7 mg/ml). A
water extract (at pH 7.0, adjusted with 0.1 mol/l
K2HPO4) from A. ursinum leaves exhibited antibacterial
activity in vitro against Listeria monocytogenes, S. aureus, E. coli, and Salmonella enterica subsp. enterica
(Sapunjieva et al. 2012). The inhibition zones were
greater in the case of Gram (?) bacteria. p
y
(
)
Ramson showed in vitro inhibitory activity on
cholesterol synthesis. Chloroform and chloroform/
acetone extracts from A. ursinum bulbs, at concentra-
tions of 166 lg/ml, inhibited cholesterol biosynthesis
by 49.3 and 48.9 %, respectively. The results were
nearly identical to those obtained for garlic extracts. Pharmacological studies ursinum bulbs, at concentra-
tions of 166 lg/ml, inhibited cholesterol biosynthesis
by 49.3 and 48.9 %, respectively. The results were
nearly identical to those obtained for garlic extracts. Of the pure investigated components present in the
extracts ajoene, methyl ajoene, 2-vinyl-4H-1,3-dithiin
and allicin were the strongest cholesterol synthesis
inhibitors, providing at the concentration of 10-3 M
the inhibition values of 69.5, 72, 58.4, and 52.2 %,
respectively (Sendl et al. 1992b). Pharmacological
studies have also revealed that chloroform and
acetone/chloroform extracts from ramson exerted
in vitro inhibitory activity on 5-lipoxygenase and
cyclooxygenase, however they were less effective
than the corresponding garlic extracts (Sendl et al. 1992a). Thrombocyte aggregation test revealed no
differences between A. ursinum and A. sativum
extracts (Sendl et al. 1992a). As was mentioned above A ursinum has been ramson was more effective than with garlic at the same
concentration (the final SBP—175 ± 1.2 mm Hg). A. ursinum decreased elevated circulating insulin con-
centration and total cholesterol level, however HDL
tended to increase. Similarly, when both garlics were
consumed at lower concentrations—0.1 % (w/w)—
systolic blood pressure readings at 10, 18, and 26 days
were significantly lower in rats consuming ramson
compared to the animals consuming common garlic. Authors concluded that these effects may be associ-
ated with high concentration of glutamyl peptides,
adenosine or phenolic compounds in ramson. They
suggested that consuming A. ursinum may result in a
greater therapeutic benefit when compared to A. sativum at a given concentration. Animal studies
demonstrated that ramson-containing diet may reduce
the size of the ischemic zone and ischemia and
reperfusion—induced arrhythmias (Rietz et al. 1993). Ramson showed in vitro inhibitory activity on
cholesterol synthesis. Chloroform and chloroform/
acetone extracts from A. ursinum bulbs, at concentra-
tions of 166 lg/ml, inhibited cholesterol biosynthesis
by 49.3 and 48.9 %, respectively. The results were
nearly identical to those obtained for garlic extracts. Of the pure investigated components present in the
extracts ajoene, methyl ajoene, 2-vinyl-4H-1,3-dithiin
and allicin were the strongest cholesterol synthesis
inhibitors, providing at the concentration of 10-3 M
the inhibition values of 69.5, 72, 58.4, and 52.2 %,
respectively (Sendl et al. 1992b). Pharmacological
studies have also revealed that chloroform and
acetone/chloroform extracts from ramson exerted
in vitro inhibitory activity on 5-lipoxygenase and
cyclooxygenase, however they were less effective
than the corresponding garlic extracts (Sendl et al. 1992a). Thrombocyte aggregation test revealed no
differences between A. ursinum and A. Pharmacological studies Modern pharmacological studies have confirmed many
of the above mentioned traditional indications of
ramson. For example, a great number of in vitro and
in vivo experiments showed that A. ursinum is a plant
with a high potential for the prevention and treatment of
cardiovascular system diseases. Different extracts
obtained from the fresh leaves of A. ursinum were
tested in vitro on human platelet aggregation. The
results showed a significant inhibitory activity of the
ethanol extract on ADP-induced aggregation. The
mechanism of action was similar to that of a reference
drug Clopidogrel (Hiyasat et al. 2009). It was suggested,
that the active compounds exerting antiaggregatory
effect are 1,2-di-O-a-linolenoyl-3-O-b-D-galactopyra-
nosyl-sn-glycerol (DLGG) (Fig. 5) and b-sitosterol
3-O-b-D-glucopyranoside (Sabha et al. 2012). DLGG
has previously been identified in a number of medicinal
and food plants, and has been shown to possess anti-
inflmmatory activity (Larsen and Christensen 2007). In vivo experiments on rats fed for 8 weeks
standard diet with 2 % of pulverized A. ursinum
leaves showed significantly lower plasma ACE activ-
ity in the ramson group as compared to control (Rietz
et al. 1993). The studies performed on Spontaneously
Hypertensive Rats (Okamoto strain) that were fed with
diet containing 1 % w/w ramson (Pfannenschmidt,
Inc. of Hamburg) showed that after 45 days it reduced
final mean systolic blood pressure when compared to
control (173 ± 0.7 vs. 189 ± 1.2 mm Hg respec-
tively) (Preuss et al. 2001). Diet enrichment with 12 3 3 Phytochem Rev (2015) 14:81–97 92 ramson was more effective than with garlic at the same
concentration (the final SBP—175 ± 1.2 mm Hg). A. ursinum decreased elevated circulating insulin con-
centration and total cholesterol level, however HDL
tended to increase. Similarly, when both garlics were
consumed at lower concentrations—0.1 % (w/w)—
systolic blood pressure readings at 10, 18, and 26 days
were significantly lower in rats consuming ramson
compared to the animals consuming common garlic. Authors concluded that these effects may be associ-
ated with high concentration of glutamyl peptides,
adenosine or phenolic compounds in ramson. They
suggested that consuming A. ursinum may result in a
greater therapeutic benefit when compared to A. sativum at a given concentration. Animal studies
demonstrated that ramson-containing diet may reduce
the size of the ischemic zone and ischemia and
reperfusion—induced arrhythmias (Rietz et al. 1993). Ramson showed in vitro inhibitory activity on
cholesterol synthesis. Chloroform and chloroform/
acetone extracts from A. Pharmacological studies The chloro-
form extract from the leaves showed inhibitory effect
on Candida albicans (MIC 312 lg/ml), as well
(Ivanova et al. 2009). The fresh water extract from
the bulbs inhibited the growth of different Candida
strains, with MIC ranging from 1 mg/ml to 4 mg/ml
depending on the particular yeast strain. The adhesion
of Candida ssp. isolates to catheters (silicone-elasto-
mer—coated latex urinary Foley catheter and PCV
Thorax catheter) was not prevented by the extract even
at the maximal concentration of 4.0 mg/ml (Chudzik
et al. 2010). The extracts prepared from fresh A. ursinum leaves collected in Romania during blossom-
ing phase inhibited the growth of Candida ssp. (C. albicans, C. famata, C. glabrata, C. krusei) at
concentrations ranging from 0.5 to 4.0 mg/ml (Bagiu
et al. 2010). specific agglutination activity (the lowest concentra-
tion which still yields a visible agglutination of a 1 %
suspension of erythrocytes) of AUAL, AUAI and
AUAII was the same (being 1.2 lg/ml). A. ursinum
lectins were more potent agglutinins than the A. sativum bulb lectins ASAI and ASAII (specific
activities being 6 and 100 lg/ml, respectively), but
less active than the garlic leaf lectin (0.2 lg/ml). The occurrence in various parts of the plant of
constituents with well-known antioxidant properties,
such as flavonoids or carotenoids, urged investigations
that would confirm ramson’s antioxidative potential. As
was shown, extracts from different parts exhibited high
free radicals scavenging activity. The antioxidant effect
of ramson leaves may be associated not only with the
presence of phenolic compounds but also with high
activity of antioxidant enzymes, like catalase and
peroxidase (11.48 ± 2.90 U/mg protein and 8.85 ±
0.19 U/mg protein, respectively), whereas in the bulbs,
with superoxide dismutase (31.43 ± 6.96 U/mg pro-
tein) (Sˇtajner et al. 2008; Sˇtajner and Szo¨llosi Varga
2003). The broad spectrum of antimicrobial activity of
Allium plants is generally associated with sulfur-
containing compounds, however our own studies have
shown that other constituents may as well contribute to
that effect, to some extent. The inhibitory activity of a
mixture of diosgenin tetrasaccharide and (25R)-spi-
rost-5,25(27)-dien-3b-ol tetrasaccharide isolated from
the bulbs against Candida glabrata and C. parapsilo-
sis was determined, with MIC values of 200 and
250 lg/ml, respectively (Sobolewska et al. 2003). Both compounds however, were ineffective against
Pseudomonas aeruginosa and A. niger at concentra-
tions up to 400 lg/ml, by the disc diffusion method. Pharmacological studies Of the pure investigated components present in the
extracts ajoene, methyl ajoene, 2-vinyl-4H-1,3-dithiin
and allicin were the strongest cholesterol synthesis
inhibitors, providing at the concentration of 10-3 M
the inhibition values of 69.5, 72, 58.4, and 52.2 %,
respectively (Sendl et al. 1992b). Pharmacological
studies have also revealed that chloroform and
acetone/chloroform extracts from ramson exerted
in vitro inhibitory activity on 5-lipoxygenase and
cyclooxygenase, however they were less effective
than the corresponding garlic extracts (Sendl et al. 1992a). Thrombocyte aggregation test revealed no
differences between A. ursinum and A. sativum
extracts (Sendl et al. 1992a). A comparative analysis of the in vitro germination
and growth inhibitory effects of the ethanol extracts
from flowers and leaves of A. ursinum against A. niger,
Botrytis cinerea, Botrytis paeoniae, Fusarium oxy-
sporum f.sp. tulipae, Penicilium gladioli and Sclero-
tina sclerotiorum showed that the flower extract
possessed the highest antifungal activity (MIC 100,
60, 70, 140, 90, and 60 lg/ml, respectively). The
authors claimed that the antifungal effects of the
extracts could be positively correlated with allicin
content: 1.946 mg allicin/ml flower extract versus
0.028 mg allicin/ml leaf extract (Parvu et al. 2011). Pure
allicin
at
concentrations
1.57–6.25 lg/ml
showed inhibitory activity against Candida, Crypto-
coccus, Trichophyton, Epidermophyton, and Micros-
porum strains (Ankri and Mirelman 1999). As was mentioned above, A. ursinum has been
valued in traditional medicine as an antimicrobial
agent used either internally or externally. There is a
substantial number of reports in which the antimicro-
bial activity of various extracts prepared from differ-
ent plant parts were tested in vitro against a wide array
of bacterial and fungal strains. Antimicrobial activity of the bulb juice of A. ursinum was correlated with storage temperature and
pH levels. Its activity against selected bacteria and
fungi decreased on storage at the temperature above
4 C and with an increase in the pH value (Tynecka
et al. 1993). The antimicrobial activity of different extracts
(acetone, chloroform, ethyl acetate, n-butanol and
water) from fresh flowers and leaves of Bulgarian
ramson was studied. Acetone extracts from both parts Comparative analysis of water and methanol
extracts from ramson herb (at the concentration range
0.16–83.7
and
0.06–35.5 mg/ml,
respectively)
showed that the latter was more active against 123 Phytochem Rev (2015) 14:81–97 93 and chloroform extract from the leaves were active
against S. aureus (MIC 625 lg/ml), while none of the
extracts inhibited the growth of E. coli. Pharmacological studies With regard to antifungal properties against Tricho-
phyton mentagrophytes and Microsporum canis the
saponin mixture was active at the concentration
400 lg/ml (Sobolewska et al. 2006). Also, the volatile oil of ramson has been tested,
however it demonstrated poor antioxidant activity
against DPPH? and ABTS? in comparison to BHT
(butylated hydroxytoluene) and Trolox. On the other
hand, in the beta-carotene-linoleic bleaching test the
oil showed an effect comparable to that of BHT
(Godevac et al. 2008). Based on these results, the
authors concluded that the antioxidant effect depends
on the method used, and also on which free radical
generator or oxidant is involved (Godevac et al. 2008). It seems therefore mandatory to employ different
analytical methods that would varying oxidation
initiators and targets. Nevertheless, some isolated ramson volatile oil
constituents have revealed promising antioxidant
properties. Diallyl disulfide increased the intracellular
content of reduced glutathione in rat red blood cells,
while diallyl sulfide enhanced the activity of antiox-
idative enzymes, and activated Nrf2 protein, what
resulted in suppression of inflammatory cytokines
(Wu et al. 2001; Kalayarasan et al. 2009). There were also some studies which evaluated the
potential of ramson against parasites. For example, the
juice from the bulbs was effective against free living
nematode Rhabditis sp., larvae of Nippostrongylus
brasiliensis, and hindered the development of Ascaris
suum eggs (Chybowski 1997). Isolated ramson’s lectins were assessed for poten-
tial inhibitory effect against HIV-1- and HIV-2-
induced cytopathicity in MT4 cells (Smeets et al. 1997). The EC50 values (the concentration required to
protect MT4 cells against cytopathicity of HIV
by 50 %) of bulbs and leaf lectins were about 3 and
5 lg/ml for HIV-1 and HIV-2, respectively. The Other
pharmacological
activities
which
were
reported for A. ursinum include in vitro cytotoxicity. Nine different extracts (chloroform, methanol, and
water) prepared by hot extraction of fresh leaves,
flowers, and flower stems were analysed in vitro against
murine cancer cell lines melanoma B16 and sarcoma 12 3 3 Phytochem Rev (2015) 14:81–97 94 XC (Trypan Blue Exclusion Test of Cell Viability)
(Sobolewska et al. 2012). The methanol extracts from
the aerial parts and the aqueous extracts from leaves and
flowers were inactive or only slightly active over the
entire concentration range (10–200 lg/ml) against both
cell lines, while the aqueous extracts from flower stems
showed no activity towards melanoma B16 cells. Pharmacological studies The
chloroform extract from flower stems showed the most
promising cytotoxic activity: at the concentration of
60 lg/ml of this extract 100 % of melanoma B16 cells
were killed after 24 h, while at the concentration of
20 lg/ml—after 48 h. In both cell lines colchicine had
an ED50 value lower than 2 lg/ml (0.5 ± 0.003—
melanoma B16; 1.5 ± 0.005—sarcoma XC) after 24 h
(Sobolewska et al. 2012). Moreover, cytotoxic activity
of a mixture of diosgenin tetrasaccharide and (25R)-
spirost-5,25(27)-dien-3b-ol tetrasaccharide on mela-
noma B16, sarcoma XC and human fibroblasts HSF
was evaluated as well. The saponin mixture was found
active against murine melanoma B16 and sarcoma XC. It exhibited 100 % effect at 2 lg/ml on both strains. It
showed no activity towards human fibroblasts HSF at
concentrations below 3 lg/ml (Sobolewska et al. 2006). disulfide, allylpropyl sulfide and allicin were identified
as allergens. All are present in various A. ursinum
preparations as well. The study on garlic-allergic
patients in Taiwan revealed that garlic C,S-lyase
(alliinase) was a major Allium sativum allergen (Kao
et al. 2004). As this enzyme showed cross-reactivity
with C,S-lyases from other species, the authors
concluded that this molecule is a common allergen
in Allium plants. The potential of A. ursinum to
enhance existing anticoagulant therapy should be
taken into consideration, as well. Even though the garlic-like odor of ramson
should enable its unambiguous identification, it
should be noted that there were some cases of fatal
poisoning by ingestion of toxic plants, the leaves of
which, due to a similar shape, were mistakenly wild-
harvested as ramson. These were, in particular,
autumn crocus (meadow saffron, Colchicum autum-
nale), the lily-of-the-valley (Convallaria majalis),
and white hellebore (Veratrum album) (Colombo
et al. 2010; Sundov et al. 2005; Gilotta and Brvar
2010; Klintschar et al. 1999). Diallyl disulfide (a component of ramson volatile
oil) inhibited the proliferation of various human
cancer cell lines, including breast, lung, colon cancers,
lymphomas and neuroblastoma. The mechanism of
action involved cell cycle arrest or apoptosis. Also,
diallyl trisulfide induced apoptosis in human prostate
cancer cell lines PC-3 and DU-145 (Lai et al. 2012). Conclusions Despite centuries of use of ramson as a substitute for
garlic (A. sativum), pharmacological studies on A. ursinum bulbs and leaves have begun fairly recently,
that is about 20 years ago. Thus, data referring to A. sativum, which is a species much valued for its
therapeutic potential are much more abundant, even
though the conclusions related to its clinical efficacy are
often inconsistent. A broad spectrum of biological
activities recorded for ramson extracts and the presence
of chemical compounds with high therapeutic potential,
makes this plant species a possible candidate for future
development as a medicinal product. Undoubtedly,
some problems that may appear are associated with
producing a uniform plant material as ramson compo-
sition is very sensitive to changes in growth conditions
what could hinder large scale production and standard-
ization.Nevertheless, it is worth noting,thatdefinitely in
recentyearsasbeenrecognizedasa valuablespice plant. References Colombo ML, Assisi F, Puppa TD et al (2010) Exposures and
intoxications after herb-induced poisoning: a retrospective
hospital-based study. J Pharm Sci Res 2(2):123–136 Andersson ME (1993) Aluminium toxicity as a factor limiting
the distribution of Allium ursinum L. Ann Bot 72:607–611 Ankri S, Mirelman D (1999) Antimicrobial properties of allicin
from garlic. Microbes Infect 1(2):125–129 Condrat D, Mosoarca C, Zamfir AD et al (2010) Qualitative and
quantitative analysis of gallic acid in Alchemilla vulgaris,
Allium ursinum, Acorus calamus and Solidago virga-aurea
by chip-electrospray ionization mass spectrometry and
high performance liquid chromatography. Cent Eur J Chem
8(3):530–535 g
Bagiu RV, Vlaicu B, Butnariu M (2010) Chemical composition
and in vitro antifungal activity screening of the Allium
ursinum L. (Liliaceae). Int J Mol Sci 13:426–1436 Benkeblia N, Lanzotti V (2007) Allium thiosulfinates: chemis-
try, biological properties and their potential utilization in
food preservation. Food 1(2):193–201 Copra-Janicijevic A, Muradic S, Huseinovic S et al (2008)
Isolation of essential oils of Allium ursinum L. from Bos-
nia. Planta Med 74:PI42 Berger F (1960) Handbuch der Drogenkunde. Wilhelm Maud-
rich/Verlag, Vienna Defelice MS (2003) Wild garlic, Allium vineale L.—little to
crow about. Weed Technol 17(4):890–895 Bierzychudek P (1982) Life histories and demography of shade-
tolerant temperate forest herbs: a review. New Phytol
90:757–776 Djurdjevic L, Dinic A, Pavlovic P et al (2004) Allelopathic
potential of Allium ursinum L. Biochem Syst Ecol
32:533–544 Bła_zewicz-Woz´niak M, Michowska A (2011) The growth,
flowering and chemical composition of leaves of three
ecotypes
of
Allium
ursinum
L. Acta
Agrobot
64(4):171–180 Eggert A (1992) Dry matter economy and reproduction of a
temperate forest spring geophyte Allium ursinum. Eco-
graphy 15:45–52 Bła_zewicz-Woz´niak M, Ke˛sik T, Michowska AE (2011)
Flowering of bear garlic (Allium ursinum L.) cultivated in
the field at varied nitrogen nutrition and mulching. Acta Sci
Pol Hortorum Cultus 10(3):133–144 g p y
Ellenberg HH (1988) Vegetation ecology of Central Europe
Cambridge University Press, Cambridge, New York Epure LI, Roman GV, Ma˘ra˘cineanu R (2011) Studies on
medicinal and aromatic plants used in the therapeutic
recepies in the Bucharest university hospital. Science
Papers UASVM, LIV, pp 304–313 Block E, Naganathan S, Putman D et al (1992) Allium chemis-
try: HPLC analysis of thiosulfinates from onion, garlic,
wild garlic (ramsoms), leek, scallion, shallot, elephant
(great-headed) garlic, chive, and chinense chive. Uniquely
high allyl to methyl ratios in some garlic samples. Adverse reactions Generally, A. ursinum is recognized as safe. However,
there is some evidence of hemolytic anemia due to
oxidative damage to erythrocytes following the con-
sumption of other Alliums by domestic and farm
animals (Munday et al. 2003). It seems that diallyl tri-
and tetrasulfides, which are highly toxic to erythro-
cytes, may be largely responsible for this effect, and
these compounds are present in ramson volatile oil, as
well. Even though there were no reports of hemolysis
associated with Allium plants consumption in humans,
certain individuals whose erythrocytes are unusually
vulnerable to oxidative damage, should consume
garlic with caution. Case reports of allergic reactions
to some garlic constituents have been also described
(Borelli et al. 2007), and such compounds as diallyl Open Access
This article is distributed under the terms of the
Creative Commons Attribution License which permits any use,
distribution, and reproduction in any medium, provided the
original author(s) and the source are credited. 12 3 123 Phytochem Rev (2015) 14:81–97 95 curing hiccups, the unusual power behind the world’s most
humble vegetable. F?W Media, Cincinnati References J Agric
Food Chem 40:2418–2430 Ernst WHO (1979) Population biology of Allium ursinum in
northern Germany. J Ecol 67:347–362 Farkas A, Molna`r R, Morschhauser T et al (2012) Variation in
nectar volume and sugar concentration of Allium ursinum
L. ssp. ucrainicum in three habitats. ScientificWorldJour-
nal 2012:138579 Bo¨hling N (2008) Eine Hypothese zur Ableitung des Namens
,,Ba¨r‘‘lauch. Ber Inst Landschafts-Pflanzeno¨kologie Univ
Hohenheim Heft 17:199–204 Friesen N, Fritsch RM, Blattner FR (2006) Phylogeny and new
intrageneric classification of Allium (Alliaceae) based on
nuclear ribosomal DNA its sequences. Aliso 22:372–395 Borelli F, Capasso R, Izzo AA (2007) Garlic (Allium sativum
L.): adverse effects and drug interactions in humans. Mol
Nutr Food Res 51:1386–1397 Fritsch RM, Keusgen M (2006) Occurrence and taxonomic
significance of cysteine sulphoxides in the genus Allium L. Fritsch RM, Keusgen M (2006) Occurrence and taxonomic
significance of cysteine sulphoxides in the genus Allium L. (Alliaceae). Phytochemistry 67:1127–1135 Boscher J, Auger J, Mandon N et al (1995) Qualitative and
quantitative comparison of volatile sulphides and flavour
precursors in different organs of some wild and cultivated
garlics. Biochem Syst Ecol 23(7/8):787–791 Gilotta I, Brvar M (2010) Accidental poisoning with Veratrum
album mistaken for wild garlic (Allium ursinum). Clin
Toxicol (Phila) 48(9):949–952 Carotenuto A, De Feo V, Fattorusso E et al (1996) The flavo-
noids of Allium ursinum. Phytochemistry 41(2):531–536 Gıˆtin L, Dinica˘ R, Parnavel R (2012) The influence of extraction
method on the apparent content of bioactive compounds in
Romanian Allium spp. leaves. Not Bot Horti Agrobot Cluj
Napoca 40(1):93–97 Celka Z (2011) Relics of cultivation in the vascular flora of
medieval West Slavic settlements and castles. Biodivers
Res Conserv 22:1–110 Chase MW, Reveal JL, Fay MF (2009) A subfamilial classifi-
cation for the expanded asparagalean families Amaryllid-
aceae, Asparagaceae and Xanthorrhoeaceae. Bot J Linn
Soc 161(2):132–136 Godevac D, Vujisic´ L, Mojovic´ M et al (2008) Evaluation of
antioxidant capacity of Allium ursinum L. volatile oil and its
effect on membrane fluidity. Food Chem 107(4):1692–1700 Govaerts R (ed) (2011) World checklist of selected plant fam-
ilies. Royal Botanic Gardens, Richmond, Surrey Chudzik B, Malm A, Rajtar B et al (2010) The fresh extracts of
Allium species as potential in vitro agents against plank-
tonic and adherent cells of Candida spp. Ann Univ Mariae
Curie-Sklodowska, DDD Pharm 23(1):73–78 Hanelt P, Bu¨ttner R (2001) Mansfeld’s encyclopedia of agri-
cultural and horticultural crops. Springer, Berlin, New York
Hegnauer R (1963) Chemotaxonomie der Pflanzen. References Birkha¨user/
Verlag, Basel Hanelt P, Bu¨ttner R (2001) Mansfeld’s encyclopedia of agri-
cultural and horticultural crops. Springer, Berlin, New York Hegnauer R (1963) Chemotaxonomie der Pflanzen. Birkha¨user/
Verlag, Basel Chybowski J (1997) Badania aktywnos´ci przeciwrobaczej
wycia˛go´w czosnkowych. Herba Pol 43(4):383–387
ˇ Hendry G (1987) The ecological significance of fructan in a
contemporary flora. New Phytol 106(Suppl.):201–216 Cˇ incˇura F, Fera´kova´ V, Ma´jovsky´ J et al (1990) Pospolite
ros´liny s´rodkowej Europy. PWRiL, Warszawa Hermy M, Honnay O, Firbank L et al (1999) An ecological
comparison between ancient and other forest plant species Clickner T (2011) A miscellany of garlic: from paying off
pyramids and scaring away tigers to inspiring courage and 12 3 3 Phytochem Rev (2015) 14:81–97 96 of Europe, and the implications for forest conservation. Biol Conserv 91(1):9–22 species (wild garlic) from Southern Africa. Tanzan J Sci
37:58–72 species (wild garlic) from Southern Africa. Tanzan J Sci
37:58–72 Macku˚ J, Krejcˇa J (1989) Atlas ros´lin leczniczych. Zakład Na-
rodowy im. Ossolin´skich, Wrocław Hiyasat B, Sabha D, Gro¨tzinger K et al (2009) Antiplatelet
activity of Allium ursinum and Allium sativum. Pharma-
cology 83(4):197–204 Madaus G (1938) Lehrbuch der Biologischen Heilmittel, vol 1. Georg Thieme/Verlag, Leipzig, pp 480–483 Ivanova A, Mikhova B, Najdenski H et al (2009) Chemical
composition and antimicrobial activity of wild garlic
Allium ursinum of Bulgarian origin. Nat Prod Commun
4(8):1059–1062 Maine Department of Agriculture, Conservation and Forestry
(2013) Maine rare plant list and rare plant fact sheets. Allium canadense. http://Maine.gov. Cited 10 Feb 2013 Jandl R, Kopeszki H, Glatzel G (1997) Effect of dense Allium
ursinum (L.) ground cover on nutrient dynamics and me-
sofauna of a Fagus sylvatica (L.) woodland. Plant Soil
189:245–255 Meier H, Reid JSG (1982) Reserve polysaccharides other than
starch in higher plants. In: Loewus FA, Tanner W (eds)
Plant carbohydrates I. Encyclopedia of plant physiology,
vol 13/A. Springer, Berlin Kalayarasan S, Prabhu PN, Sriram N et al (2009) Diallyl sulfide
enhances antioxidants and inhibits inflammation through
the activation of Nrf2 against gentamicin-induced nephro-
toxicity in Wistar rats. Eur J Pharmacol 606(1–3):162–171 Meyer FG, Emmart Trueblood E, Heller JL (1999) The great
herbal of Leonhart Fuchs. De historia stripium commentari
insignes, 1542. Stanford University Press, Stanford g
g
p
ity in Wistar rats. References Eur J Pharmacol 606(1–3):162–171 Morschhauser T, Rudolf K, Botta-Duka´t Z et al (2009) Density-
dependence in the establishment of juvenile Allium ursi-
num individuals in a monodominant stand of conspecific
adults. Acta Oecol 35(5):621–629 Kao S-H, Hsu C-H, Su S-N et al (2004) Identification and
immunologic characterization of an allergen, alliin lyase,
from garlic (Allium sativum). J Allergy Clin Immunol
113(1):161–168 Munday R, Munday JS, Munday CM (2003) Comparative
effects of mono-, di-, tri-, and tetrasulfides derived from
plants of the Allium family: redox cycling in vitro and
hemolytic activity and phase 2 enzyme induction in vivo. Free Radic Biol Med 34(9):1200–1211 Karpaviciene B (2006) Distribution of Allium ursinum L. in
Lithuania. Acta Biol Univ Daugavp 6(1–2):117–122 Keusgen M, Ju¨nger M, Krest I et al (2003) Development of
biosensor specific for cysteine sulfoxides. Biosens Bio-
electron 18(5–6):805–812 Nagori BP, Solanki R, Sharma N (2010) Natural healing
agent: garlic, an approach to healthy life. IJRAP 1(2):
358–366 Klintschar M, Beham-Schmidt C, Radner H et al (1999) Col-
chicine poisoning by accidental ingestion of meadow saf-
fron (Colchicum autumnale): pathological and medicolegal
aspects. Forensic Sci Int 106(3):191–200 Oborny B, Botta-Dukat Z, Rudolf K, Morschhauser T (2011)
Population ecology of Allium ursinum, a space-monopo-
lizing clonal plant. Acta Bot Hung 53(3–4):371–388 Kovacs JA (2007) Data to vegetation biology and coenological
relations of Allium ursinum L. stands in Eastern Transyl-
vania. Kanitzia 15:63–76 Oszmian´ski J, Kolniak-Ostek J, Wojdyło A (2013) Character-
ization and content of flavonol derivatives of Allium ursi-
num L. plant. J Agric Food Chem 61:176–184 Kubec R, Svobodova M, Velisek J (2000) Distribution of
S-alk(en)ylcysteine sulfoxides in some Allium species. Identification of a new flavour precursor: S-ethylcysteine
sulfoxide (ethiin). J Agric Food Chem 48:428–433 num L. plant. J Agric Food Chem 61:176–184 Parvu M, Parvu AE, Vlase L et al (2011) Antifungal properties
of Allium ursinum L. ethanol extract. J Med Plants Res
5(10):2041–2046 Kubiak-Martens L (2002) New evidence for the use of root
foods in pre-agrarian subsistence recovered from the late
Mesolithic site at Halsskov, Denmark. Veg Hist Archaeo-
bot 11:23–31 Preuss HG, Clouatre D, Mohamadi A et al (2001) Wild garlic
has a greater effect than regular garlic on blood pressure
and blood chemistries of rats. References 1) 224, B020 Sundov Z, Nincevic Z, Definis-Gojanovic M et al (2005) Fatal
colchicine poisoning by accidental ingestion of meadow
saffron—case report. Forensic Sci Int 149(2–3):253–256 Schmitt B, Schulz H, Strosberg J et al (2005) Chemical char-
acterization of Allium ursinum L. depending on harvesting
time. J Agric Food Chem 53:7288–7294 Synowiec A, Gniewosz M, Zieja I et al (2010) Poro´wnanie
włas´ciwos´ci
przeciwdrobnoustrojowych
ekstrakto´w
z
czosnku niedz´wiedziego (Allium ursinum). Zesz Probl
Poste˛po´w Nauk Rol 553:203–209 Sendl A (1995) Allium sativum and Allium ursinum: part 1. Chemistry,
analysis,
history,
botany. Phytomedicine
1(4):323–329 Szafer W, Zarzycki W (1972) Szata ros´linna Polski. PWN,
Warszawa Szafer W, Kulczyn´ski S, Pawłowski B (1988) Ros´liny polskie. PWN, Warszawa Sendl A, Wagner H (1991) Isolation and identification of
homologues of ajoene and alliin from bulb-extracts of
Allium ursinum. Planta Med 57(4):361–362 Treben M (1992) Apteka Pana Boga. Porady i praktyka sto-
sowania zio´ł leczniczych. Natur-Produkt TOM-MARK,
Warszawa Sendl A, Elbl G, Steinke B et al (1992a) Comparative phar-
macological investigations of Allium ursinum and Allium
sativum. Planta Med 58(1):1–7 Tynecka Z, Szczes´niak Z, Głowniak K (1993) The effect of
various environmental conditions on the antimicrobial
activity of Allium ursinum. Planta Med 59(7 SUPPL):A701 Sendl A, Schliack M, Lo¨ser R et al (1992b) Inhibition of cho-
lesterol synthesis in vitro by extracts and isolated com-
pounds
prepared
from
garlic
and
wild
garlic. Atherosclerosis 94(1):79–85 Wagner H, Sendl A (1990) Barlauch und Knoblauch. Verg-
leichende chemische und pharmakologische Untersuch-
ungen von Barlauch- und Knoblauchextrakten. Dtsch
Apoth Ztg 33:1809–1815 Shmanova IW, Krichfalushii WW (1995) Biomorphological
and ecologo-coenotic characteristics of Allium ursinum L. in Carpathians. Rast Res 31(3):1–18 Wiater M, Sobolewska D, Janeczko Z (1998) Fatty acids in lipid
fraction from the undeground parts of Allium ursinum L. In:
XVII Naukowy Zjazd Polskiego Towarzystwa Farm-
aceutycznego
‘‘Farmacja
w
perspektywie
XXI
w.’’. Streszczenia. Krako´w. 10–13 Sept 1998 Smeets K, Van Damme EJ, Van Leuven F et al (1997) Isolation,
characterization and molecular cloning of a leaf-specific
lectin from ramsons (Allium ursinum L.). Plant Mol Biol
35(4):531–535 Sobolewska D, Janeczko Z, Galanty A, Trojanowska D (2003)
Cytotoxic, antifungal and antibacterial activity of spirost-
anol saponin from ramson Allium ursinum L. In: Pro-
ceedings of the 3rd international symposium on natural
drugs. Naples. 2–4 Oct 2003 Wu CC, Sheen LY, Chen HW et al (2001) Effects of organo-
sulfur compounds from garlic oil on the antioxidation
system in rat liver and red blood cells. References Int Urol Nephrol 32(4):
525–530 Lai KC, Kuo CL, Ho HC et al (2012) Diallyl sulfide, diallyl
disulfide and diallyl trisulfide affect drug resistant gene
expression in colo 205 human colon cancer cells in vitro
and in vivo. Phytomedicine 19:625–630 Puxbaum H, Ko¨nig G (1997) Observation of dipropenyl disul-
fide and other organic sulfur compounds in the atmosphere
of a beech forest with Allium ursinum ground cover. Atmos
Environ 31(2):291–294 Landshuter J, Lohmu¨ller EM, Knobloch K (1994) Purification
and characterization of C-S-lyase from ramson, the wild
garlic, Allium ursinum. Planta Med 60:343–347 Rejewski M (1996) Pochodzenie łacin´skich nazw ros´lin polsk-
ich. Przewodnik botaniczny. Ksia˛_zka i Wiedza, Warszawa Richter T (1999) Ba¨rlauch in Medizin und Mythologie. Pharm
Ztg 144:2197–2198 Larsen E, Christensen LP (2007) Common vegetables and fruits
as a source of 1,2-di-O-a-linolenoyl-3-O-b-D-galactopyr-
anosyl-sn-glycerol. A potential anti-inflammatory and
antitumor agent. J Food Lipids 14:272–279 Rietz B, Isensee H, Strobach H et al (1993) Cardioprotective
actions of wild garlic (Allium ursinum) in ischemia and
reperfusion. Mol Cell Biochem 119:143–150 Leporatti ML, Ivancheva S (2003) Preliminary comparative
analysis of medicinal plants used in the traditional medi-
cine of Bulgaria and Italy. J Ethnopharmacol 87:123–142 Rola K (2012) Taxonomy and distribution of Allium ursinum
(Liliaceae) in Poland and adjacent countries. Biologia
67(6):1080–1087 Leuschner C, Lendzion J (2009) Air humidity, soil moisture and
soil chemistry as determinants of the herb layer composi-
tion in European beech forests. J Veg Sci 20:288–298 Rychnovska´ M, Bedna´rˇ V (1998) Floodplain forest: herb layer
as indicator of its ecological status. Acta Univ Palacki
Olomuc Fac Rer Nat Biol 36:7–15 Lyantagaye SL (2011) Ethnopharmacological and phytochem-
ical review of Allium species (sweet garlic) and Tulbaghia Sabha D, Hiyasat B, Gro¨tzinger K et al (2012) Allium ursinum
L.: bioassay-guided isolation and identification of a 12 3 Phytochem Rev (2015) 14:81–97 97 galactolipid and a phytosterol exerting antiaggregatory
effects. Pharmacology 89(5–6):260–269 galactolipid and a phytosterol exerting antiaggregatory
effects. Pharmacology 89(5–6):260–269 Sˇtajner D, Popovic´ BM, Candanovic´-Brunet J et al (2008)
Antioxidant and scavenger activities of Allium ursinum. Fitoterapia 79(4):303–305 Sapunjieva T, Alexieva I, Mihaylova D et al (2012) Antimi-
crobial and antioxidant activity of extracts of Allium ursi-
num L. J Biol Sci Biotech SE/ONLINE:143–145 Strzelecka H, Kowalski J (eds) (2000) Encyklopedia Zielarstwa
i Ziołolecznictwa. PWN, Warszawa Schmitt B, Glodek J, Keusgen M (2002) Ontogenic changes of
cysteine sulphoxides in Allium ursinum L. Revista de
Fitoterapia 2(Suppl. References Food Chem Toxicol
39(6):563–569 Wu H, Dushenkov S, Ho CT et al (2009) Novel acetylated fla-
vonoid glycosides from the leaves of Allium ursinum. Food
Chem 115(2):592–595 Sobolewska D, Janeczko Z, Kisiel W et al (2006) Steroidal
glycosides from the underground parts of Allium ursinum
L. and their cytostatic and antimicrobial activity. Acta Pol
Pharm Drug Res 63(3):219–223 Yoo KS, Pike LM (1998) Determination of flavor precursor
compound S-alk(en)yl-L-cysteine sulfoxides by an HPLC
method and their distribution in Allium species. Sci Hortic
75(1–2):1–10 Sobolewska D, Janeczko Z, Podolak I et al (2009) Densitometric
analysis of diosgenin in methanolic extracts of Allium
ursinum collected in different periods of plant develop-
ment. J Planar Chromatogr 22(4):305–307 Zarzycki K, Mirek Z (2006) Red list of plants and fungi in
Poland. Czerwona lista ros´lin i grzybo´w Polski. Instytut
Botaniki im. W. Szafera PAN, Krako´w Sobolewska D, Galanty A, Michalik M (2012) Preliminary
evaluation of cytotoxic activity of Allium ursinum extracts. Czas Aptek 12(228):41–44 Sˇtajner D, Szo¨llosi Varga I (2003) An evaluation of the anti-
oxidant abilities of Allium species. Acta Biol Szeged
47(1–4):103–106 123
|
https://openalex.org/W2104706617
|
https://europepmc.org/articles/pmc3399817?pdf=render
|
English
| null |
BAY61-3606 Affects the Viability of Colon Cancer Cells in a Genotype-Directed Manner
|
PloS one
| 2,012
|
cc-by
| 9,312
|
Abstract Background: K-RAS mutation poses a particularly difficult problem for cancer therapy. Activating mutations in K-RAS are
common in cancers of the lung, pancreas, and colon and are associated with poor response to therapy. As such, targeted
therapies that abrogate K-RAS-induced oncogenicity would be of tremendous value. Methods: We searched for small molecule kinase inhibitors that preferentially affect the growth of colorectal cancer cells
expressing mutant K-RAS. The mechanism of action of one inhibitor was explored using chemical and genetic approaches. Results: We identified BAY61-3606 as an inhibitor of proliferation in colorectal cancer cells expressing mutant forms of K-
RAS, but not in isogenic cells expressing wild-type K-RAS. In addition to its anti-proliferative effects in mutant cells, BAY61-
3606 exhibited a distinct biological property in wild-type cells in that it conferred sensitivity to inhibition of RAF. In this
context, BAY61-3606 acted by inhibiting MAP4K2 (GCK), which normally activates NFkb signaling in wild-type cells in
response to inhibition of RAF. As a result of MAP4K2 inhibition, wild-type cells became sensitive to AZ-628, a RAF inhibitor,
when also treated with BAY61-3606. Conclusions: These studies indicate that BAY61-3606 exerts distinct biological activities in different genetic contexts. se studies indicate that BAY61-3606 exerts distinct biological activities in different genetic contexts. Conclusions: These studies indicate that BAY61-3606 exerts distinct biological activities in different gen Citation: Lau KS, Zhang T, Kendall KR, Lauffenburger D, Gray NS, et al. (2012) BAY61-3606 Affects the Viability of Colon Cancer Cells in a Genotype-Directed
Manner. PLoS ONE 7(7): e41343. doi:10.1371/journal.pone.0041343
Editor: David J. Reiner, University of North Carolina, United States of America Citation: Lau KS, Zhang T, Kendall KR, Lauffenburger D, Gray NS, et al. (2012) BAY61-3606 Affects the Viability of Colon Cancer Cells in a Genotype-Directed
Manner. PLoS ONE 7(7): e41343. doi:10.1371/journal.pone.0041343 Editor: David J. Reiner, University of North Carolina, United States of America Received February 2, 2012; Accepted June 20, 2012; Published July 18, 2012 Copyright: 2012 Lau et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License,
use, distribution, and reproduction in any medium, provided the original author and source are credited. et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted
roduction in any medium, provided the original author and source are credited. Abstract Funding: This work was supported by grants from the National Cancer Institute (K01-CA118425 and P30-CA006516) and the American Cancer Society (MGO-
114877) and a donation from the Merlino Family Endowment Fund. KSL is a Robert Black Fellow of the Damon Runyon Cancer Research Foundation. The funders
had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: khaigis@partners.org via inactivation of its GTPase activity, direct pharmacologic
inhibition of RAS is not a viable therapeutic strategy. An alternate
strategy to counteract mutant K-RAS is to inhibit its downstream
effectors, for example, the RAF-MEK-ERK (MAPK) pathway. MEK inhibitors have received attention due their allosteric
mechanism of action, which confers extreme specificity, and their
demonstrated efficacy in melanomas and colon cancers expressing
activated B-RAF [6,7]. MEK inhibitors perform poorly in cancers
expressing mutant K-RAS, however, perhaps due to secondary
mutations that affect response or the existence of MEK-
independent signaling downstream of RAF [8,9]. Given the
presumptiveness of these scenarios, it is clear that a better
understanding of how the K-RAS signaling network operates in
cancer is needed to develop novel therapies. Ken S. Lau1,2, Tinghu Zhang3, Krystle R. Kendall1, Douglas Lauffenburger2, Nathanael S. Gray3,
Kevin M. Haigis1* ,2, Tinghu Zhang3, Krystle R. Kendall1, Douglas Lauffenburger2, Nathanael S. Gray
1 1 Molecular Pathology Unit, Center for Cancer Research and Center for Systems Biology, Massachusetts General Hospital, Charlestown, Massachusetts, United States of
America, 2 Department of Biological Engineering and Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts,
United States of America, 3 Department of Cancer Biology, Dana Farber Cancer Institute, Boston, Massachusetts, United States of America PLoS ONE | www.plosone.org Cell cycle analysis and cell viability assays Cell cycle analysis was performed via FACS-based propidium
iodide quantification, using standard methods. To measure cell
viability, cells were grown in 96-well plates in the presence or
absence of drug for 72 hours, fixed with 4% paraformaldehyde,
and then stained with Syto60 (Invitrogen). Plates were imaged and
quantified using the LiCor Odyssey system (LiCor). Introduction RAS family GTPases act as binary switches that undergo
a conformational change upon binding to GTP, allowing them to
engage a host of downstream signaling effectors including RAF,
PI3K, and RALGDS [1]. The intrinsic GTPase activity of RAS
hydrolyzes GTP to GDP, with the help of GTPase activating
protein (GAP) cofactors, to inactivate its signaling capability. Missense mutations in codons 12, 13, 61, or 146 are common in
cancer and are associated with resistance to GAP activity, allowing
RAS to persist in the activated, GTP-bound state. Activating K-
RAS mutations occur in approximately 15% of all cancers (making
it one of the most commonly mutated oncogenes), but are
particularly common in the most lethal forms of cancer, such as
those arising in the biliary tract, colon, lung, and pancreas [2]. In
colorectal cancer, for example, K-RAS is mutated in nearly 40%
of cases [2]. Importantly, tumors with K-RAS mutations are
especially refractory to conventional and targeted therapies and
are usually associated with poor prognosis [3–5]. In recent years, large-scale functional genomic approaches have
been employed to discover kinase targets that when knocked down
are ‘‘synthetic lethal’’ with mutant RAS. Potential therapeutic
targets that have been identified include TBK1 [10], STK33 [11],
CDK4 [12], and PLK1 [13], although it remains to be seen
whether any of these represent bona fide therapeutic targets for K-
RAS mutant cancers. Whereas understanding the mechanisms by The main challenge of counteracting the oncogenic effects of
activated K-RAS is the inability to directly inhibit the mutant
protein. Because the signaling properties of K-RAS are enhanced July 2012 | Volume 7 | Issue 7 | e41343 PLoS ONE | www.plosone.org 1 Selective Activity of BAY61-3606 control. All signals were normalized to a common control cell line
lysate in order for assays between plates to be comparable. control. All signals were normalized to a common control cell line
lysate in order for assays between plates to be comparable. which K-RAS signals through these targets is central to the design
of effective drugs, a less studied, and often overlooked, question is
why wild-type cells, which also express these targets, tolerate loss of
function of these enzymes. This issue is equally important for drug
design
because
the
advantage
of
targeted
therapies
(over
conventional chemotherapies) is their potential selectivity for
malignant cells. AZ-628 and BAY61-3606 suppress growth in cells
expressing K-RASG13D In an effort to identify novel therapeutic targets for colorectal
cancers expressing mutant K-RAS, we performed a screen for
small molecule kinase inhibitors that affect viability in a genotype-
specific manner. In these studies, we utilized a set of isogenic colon
cancer cell lines that differ only in their K-RAS mutation status. The parental cell lines, HCT-116 and DLD-1, carry a heterozy-
gous activating mutation in K-RAS (G13D/+). The derivative cell
lines, HKe-3 and DKs-8, retain the wild-type allele, but have lost
the mutant allele of K-RAS by virtue of gene targeting [17]. Consistent with our previous work [9], we found that K-RAS
mutant cells were hypersensitive to AZ-628, a pan-RAF inhibitor
[18,19], compared to wild-type cells, but insensitive to CI-1040,
a MEK inhibitor [6] (Fig. 1a). We also found that BAY61-3606,
a Spleen Tyrosine Kinase (SYK) inhibitor, affected overall viability
in HCT-116 and DLD-1 cells compared to HKe-3 and DKs-8
cells (Fig. 1a, Fig. S1a). Chemical derivation of BAY61-3603 Details on the synthesis of BAY derivatives, and the structures of
those derivatives, can be found in Figure S5. Results AZ-628 and BAY61-3606 suppress growth in cells
expressing K-RASG13D Methods Cell lines, knockdowns, and drug treatments
All colon cancer cell lines were maintained in DMEM
supplemented
with
penicillin
(100 units/mL),
streptomycin
(100 mg/mL), and 10% fetal bovine serum (FBS). The rectal
cancer cell line (Car1) was maintained in DMEM/F12 supple-
mented with penicillin (100 units/mL)/streptomycin (50 mg/mL),
and 5% FBS. Knockdowns were achieved with pSICOR or
pLKO lentiviral vectors [16]. The target sequences for knock-
downs can be found in Table S2. In drug treatment experiments,
cells were plated for 24 hours prior to exposure to drug. AZ-628
was obtained from AstraZeneca. CI-1040 was obtained from
Pfizer. R406 was synthesized in the Gray laboratory. BAY61-3606
and IKK VII were purchased from EMD Biosciences. BAY
derivatives were synthesized for this study. To determine how BAY61-3606 affected viability of cells
expressing mutant K-RAS, we analyzed the cell cycle of cells
treated with the drug. We found that BAY61-3606 did not alter
the cell cycle profile of DLD-1 cells, nor did it induce apoptosis
(Fig. 1b). In contrast to colorectal cancer cells expressing mutant
K-RAS, cell lines that express mutant B-RAF were sensitive to
inhibition of MEK and treatment with CI-1040 induced G1 arrest
[6] (Fig. 1b, c). Interestingly, we found that B-RAF mutant cells
lines were also sensitive to BAY61-3606 and that CI-1040 and
BAY61-3606 cooperated to produce an enhanced negative effect
on viability of cells expressing activated B-RAF (Fig. 1c). Taken
together, these observations suggested (1) that BAY61-3606
inhibits a protein that is required for overall viability in cells
expressing mutant K-RAS or B-RAF and (2) that BAY61-3606
targets a pathway that is independent of canonical MAPK/ERK
signaling. Biochemical activity assays The biochemical activity of BAY61-3606 and derivatives were
measured in two ways. First, we used Ambit’s KINOMEscanTM
technology to identify those kinases that are inhibited for substrate
binding by the compounds, all assayed at 1 mM. Second, we used
Invitrogen’s SelectScreenH Biochemical Kinase Profiling Service
to determine the in vitro IC50s for the compounds against specific
kinases. g
In this study, we have characterized the activity of BAY61-3606
in the context of colorectal cancer, providing insight into (1)
potential therapeutic targets for cancers expressing mutant K-RAS
and (2) pathways that regulate the response of non-mutant cells to
targeted inhibitors. BAY61-3606 was originally identified as an
orally available, ATP-competitive inhibitor of Spleen Tyrosine
Kinase (SYK) [14]. Since SYK plays an active role in in-
flammatory response, BAY61-3606 has mainly been used for
studying immune cell function. For example, BAY61-3606
suppresses antigen-induced airway inflammation in rats and B
cell migration in mice [14,15]. While all of the effects of BAY61-
3606 in immune cells are linked to its ability to inhibit SYK, it is
unknown whether BAY61-3606 has alternate targets of biological
relevance in other cellular contexts. In this study, we have
characterized two SYK-independent activities associated with
BAY61-3606 in colorectal cancer cells. Bio-Plex signaling assays The Bio-Plex assay system was used to measure signaling in
drug-treated cells. Briefly, cells were incubated in the presence of
drug for various amounts of time and then lysed in Bio-Rad cell
lysis buffer (Bio-Rad). Protein quantification was performed using
BCA assay (Pierce) and 5 mg of protein from each sample was used
for Bio-Plex analysis. Phospho-signaling assays were performed
using available phospho-signaling assay kits and quantified on
a Bio-Plex 200 system (Bio-Rad): p-Ikba (Ser32/Ser36), p-JNK
(Thr183/Tyr185),
p-MEK1
(Ser217/Ser221),
p-ERK1/2
(Thr202/Tyr204,
Thr185/Tyr187),
p-p90RSK
(Thr359/
Ser363), p-p38 (Thr180/Tyr182), p-c-JUN (Ser63), p-ATF2
(Thr71),
p-AKT
(Ser473),
p-S6
(Ser235/Ser236),
p-STAT3
(Ser727), p-STAT3 (Tyr705), and p-GSK3a/b (Ser21/Ser9). Bio-Plex assay for total MEK1 was also performed as a loading To test the second part of this hypothesis directly, we measured
the activation state of MEK and ERK in cells that were treated
with BAY61-3606. We found that inhibition of RAF with AZ-628
suppressed MEK and ERK phosphorylation, but that BAY61-
3606 was unable to do so (Fig. 1d). Thus, while AZ-628 and
BAY61-3606 both selectively affect viability in K-RAS mutant
cells, they appear to do so through distinct pathways, with BAY61-
3606 targeting a pathway that is independent of MEK and ERK. July 2012 | Volume 7 | Issue 7 | e41343 PLoS ONE | www.plosone.org 2 Figure 1. BAY61-3606 affects viability in cells expressing mutant K-RAS or B-RAF through a MAPK-independent pathway. (a) Cell
viability quantified by Syto60 after 72 hours of AZ-628, CI-1040 or BAY61-3606 treatment in HCT-116 (K-RASG13D/+, red) or HKe-3 (K-RAS2/+, blue) cell
lines. Relative cell viability was normalized to DMSO vehicle treated control for each cell line. Error bars represent SEM for 3 independent experiments. Cells expressing mutant K-RAS were relatively sensitive to AZ-628 and BAY61-3606, but not CI-1040. (b) Cell cycle profiles, as determined by
propidium iodide staining, of colorectal cancer cells with mutant B-RAF (HT-29) treated with CI-1040 or mutant K-RAS (DLD-1) treated with BAY61-
3606. While inhibition of MEK induces G1 arrest in HT-29 cells, as evidenced by the loss of the 4N peak, BAY61-3606 did not appear to alter the profile
of DLD-1 cells. (c) Cell viability of colorectal cancer cells expressing mutant B-RAF (V600E) after 72 hours of treatment with BAY61-3606 and/or CI-
1040. Cells expressing mutant B-RAF (HT-29 – solid outline and RKO – dotted outline) were sensitive to both BAY61-3606 and CI-1040 and these two
inhibitors cooperated to produce an enhanced response. Bio-Plex signaling assays (d) Phospho-MEK1 (Ser217/Ser221) and phospho-ERK1/2 (Thr202/Tyr204, Thr185/Tyr187)
levels in K-RAS wild-type and mutant cells after 45 minute exposure to 1 mM AZ-628, 1 mM BAY61-3606, or vehicle control. Signals were measured
using Bio-Plex assays. Relative signal was normalized to a master control lysate. AZ-628 treatment reduced the level of phospho-MEK and phospho-
ERK, but BAY61-3606 did not. doi:10.1371/journal.pone.0041343.g001
Selective Activity of BAY61-3606 Selective Activity of BAY61-3606 Figure 1. BAY61-3606 affects viability in cells expressing mutant K-RAS or B-RAF through a MAPK-independent pathway. (a) Cell
viability quantified by Syto60 after 72 hours of AZ-628, CI-1040 or BAY61-3606 treatment in HCT-116 (K-RASG13D/+, red) or HKe-3 (K-RAS2/+, blue) cell
lines. Relative cell viability was normalized to DMSO vehicle treated control for each cell line. Error bars represent SEM for 3 independent experiments. Cells expressing mutant K-RAS were relatively sensitive to AZ-628 and BAY61-3606, but not CI-1040. (b) Cell cycle profiles, as determined by
propidium iodide staining, of colorectal cancer cells with mutant B-RAF (HT-29) treated with CI-1040 or mutant K-RAS (DLD-1) treated with BAY61-
3606. While inhibition of MEK induces G1 arrest in HT-29 cells, as evidenced by the loss of the 4N peak, BAY61-3606 did not appear to alter the profile
of DLD-1 cells. (c) Cell viability of colorectal cancer cells expressing mutant B-RAF (V600E) after 72 hours of treatment with BAY61-3606 and/or CI-
1040. Cells expressing mutant B-RAF (HT-29 – solid outline and RKO – dotted outline) were sensitive to both BAY61-3606 and CI-1040 and these two
inhibitors cooperated to produce an enhanced response. (d) Phospho-MEK1 (Ser217/Ser221) and phospho-ERK1/2 (Thr202/Tyr204, Thr185/Tyr187)
levels in K-RAS wild-type and mutant cells after 45 minute exposure to 1 mM AZ-628, 1 mM BAY61-3606, or vehicle control. Signals were measured
using Bio-Plex assays. Relative signal was normalized to a master control lysate. AZ-628 treatment reduced the level of phospho-MEK and phospho-
ERK, but BAY61-3606 did not. d i 10 1371/j
l
0041343 001 doi:10.1371/journal.pone.0041343.g001 doi:10.1371/journal.pone.0041343.g001 (Fig. 2a). Knockdown of SYK did not affect overall growth rate
nor did it significantly affect the response of DLD-1 cells to
BAY61-3606 (Fig. 2b). Moreover, treatment of cells with R406,
a structurally distinct inhibitor of SYK, did not result in
a preferential effect in cells expressing mutant K-RAS (Fig. 2b). Taken together, these results suggested that SYK was not the
relevant target of BAY61-3606 in colorectal cancer cells expressing
mutant K-RAS. PLoS ONE | www.plosone.org Identification of biologically active BAY61-3606
derivatives Secondary to our genetic analysis, we took a chemical approach
to studying BAY61-3606. In this study, we generated 30
structurally related derivatives of BAY61-3606 and assayed their
ability to selectively affect viability of cells expressing mutant K-
RAS (Fig. S2). Of these 30 derivatives, only one (derivative 6)
retained its selectivity with potency similar to the parent
compound (Fig. 4a, b, Fig. S1b). Others (e.g. derivative 8) retained
selectivity but lost potency. To address the selectivity of BAY
derivative 6 more broadly, we assessed its activity in a panel of
colorectal cancer cell lines that were either mutant for wild-type
for K-RAS. Consistent with our analysis of isogenic pairs, 4/5 cell
lines expressing mutationally activated K-RAS responded to BAY
derivative 6, while 0/3 cell lines expressing wild-type K-RAS
responded (Fig. S3). Figure 2. Inhibition of SYK is not responsible for the BAY61-
3606 effect on cell viability in colorectal cancer cells. (a)
Knockdown of SYK protein in DLD-1 cells via shRNA, as shown by
Western Blotting. The Ramos (R) cell line, a hematopoetic cell line with
a high expression of SYK, was used as a positive control. (b) Cell viability
of DLD-1 cells (red), and its K-RAS wild-type (DKs-8 – blue) and SYK
knockdown (red shaded) derivatives after 72 hours of treatment with
1 mM of BAY61-3606 or R406, a distinct SYK inhibitor. Relative cell
viability was normalized to DMSO vehicle treated control for each cell
line. Error bars represent SEM for 3 independent experiments. DLD-1
cells and its K-RAS wild-type derivative did not exhibit sensitivity to
R406, while knocking down SYK in DLD-1 cells only minimally affected
its sensitivity to BAY61-3606. To compare with BAY61-3606, we assayed the ability of several
derivatives to competitively inhibit active site binding in a large
panel of kinases. Remarkably, each of the derivatives that were
tested essentially lost their ability to competitively inhibit active site
binding of the 402 kinases that were analyzed, including HIPK2
(Fig. S4). Consistent with this observation, in vitro activity assays
revealed a loss of kinase inhibition activity for the two derivatives
tested (Fig. S5). SYK is not the target of BAY61-3606 in cells expressing
mutant K-RAS SYK is not the target of BAY61-3606 in cells expressing
mutant K-RAS Given that BAY61-3606 was originally developed as an ATP-
competitive inhibitor of SYK [14], it was not surprising that it
appeared to target a MEK/ERK-independent pathway. Never-
theless, HCT-116 cells, which are sensitive to BAY61-3606, do not
express detectable levels of SYK, suggesting that it may not be the
target of BAY61-3606 in this context. To explore this further, we
used lentiviral shRNA to knock down SYK in DLD-1 cells July 2012 | Volume 7 | Issue 7 | e41343 PLoS ONE | www.plosone.org 3 Selective Activity of BAY61-3606 Figure 2. Inhibition of SYK is not responsible for the BAY61-
3606 effect on cell viability in colorectal cancer cells. (a)
Knockdown of SYK protein in DLD-1 cells via shRNA, as shown by
Western Blotting. The Ramos (R) cell line, a hematopoetic cell line with
a high expression of SYK, was used as a positive control. (b) Cell viability
of DLD-1 cells (red), and its K-RAS wild-type (DKs-8 – blue) and SYK
knockdown (red shaded) derivatives after 72 hours of treatment with
1 mM of BAY61-3606 or R406, a distinct SYK inhibitor. Relative cell
viability was normalized to DMSO vehicle treated control for each cell
line. Error bars represent SEM for 3 independent experiments. DLD-1
cells and its K-RAS wild-type derivative did not exhibit sensitivity to
R406, while knocking down SYK in DLD-1 cells only minimally affected
its sensitivity to BAY61-3606. doi:10 1371/journal pone 0041343 g002 could only identify a single shRNA sequence that produced
adequate knockdown of this target. As a result, this study was
suggestive, but not conclusive, of a role for HIPK2 as a target of
BAY61-3606 in cells expressing mutant K-RAS. We sought an
independent way to identify the target of BAY61-3606. Identification of biologically active BAY61-3606
derivatives BAY61-3606 targets MAP4K2 to affect the response of
wild-type cells to AZ-628 y
doi:10.1371/journal.pone.0041343.g002 y
In analyzing the relative activities of AZ-628 and BAY61-3606,
we tested whether these two compounds would cooperate to
produce an enhanced effect in cells expressing mutant K-RAS,
similar to the cooperative effect seen with CI-1040 and BAY61-
3606 in cells expressing mutant B-RAF. AZ-628 and BAY61-3606
did not cooperate in HCT-116 cells (Fig. 5a), however, suggesting
that they may target a common pathway. Interestingly these two
inhibitors did cooperate to negatively affect viability of cells
expressing wild-type K-RAS. In essence, HKe-3 cells that were
formally resistant to AZ-628 became sensitive when they were also
treated with BAY61-3606 (Fig. 5a). BAY derivative 6, which
retained its ability to selectively suppress proliferation in cells
expressing mutant K-RAS (Fig. 4b), lost its ability to cooperate
with AZ-628 in cells expressing wild-type K-RAS (Fig. 5b). BAY61-3606 targets a small number of kinases We hypothesized that the activity of BAY61-3606 in colon
cancer cells was due to an ‘‘off-target’’ effect of the inhibitor, yet
little
was known about the promiscuity of this particular
compound. To identify other potential targets of BAY61-3606,
we first assayed the ability of BAY61-3606 to competitively inhibit
active site binding in a panel of 402 kinases. We found that 1 mM
BAY61-3606 inhibited binding by greater than 90% for only 15
kinases (Fig. 3a, Table 1, Table S1), indicating that it is a highly
selective inhibitor, similar in selectivity to two clinical kinase
inhibitors, Imatinib and Gefitinib [20]. Since inhibition of active
site binding may, or may not, be directly correlated with inhibition
of kinase activity, we performed a secondary analysis in which we
determined the in vitro IC50 values for BAY61-3606 against many
of these candidate targets. This study revealed MAP4K2, a STE-
20 family kinase, as the kinase most sensitive to BAY61-3606, with
an in vitro IC50 of 11.3 nM (Table 1). With the exception of SYK, BAY61-3606 was more than 10-
fold more effective at inhibiting MAP4K2 than any other kinase. Moreover, BAY derivative 6 lost its ability to effectively inhibit the
in vitro kinase activity of MAP4K2 (Fig. S5). Based on these
observations, we explored whether MAP4K2 plays a role in the
sensitivity of wild-type and K-RAS mutant cells to BAY61-3606. HKe-3 and Dks-8 cells lacking MAP4K2 were hypersensitive to
AZ-628, while MAP4K2 knockout had no effect on the response
of HCT-116 or DLD-1 cells (Fig. 5c, Fig. S6a). We reasoned that
if inhibition of MAP4K2 by BAY61-3606 accounted for the
altered response of wild-type cells to AZ-628, then K-RAS wild-
type cells lacking MAP4K2 would not be further sensitized to AZ-
628 by treatment with BAY61-3606. As predicted, BAY61-3606
and AZ-628 failed to cooperatively affect viability in HKe-3 cells
lacking MAP4K2 (Fig. 5d). The results strongly suggest that enetic analysis of potential BAY61-3606 targets To follow-up on our biochemical analysis of BAY61-3606
activity,
we
used
lentiviral
shRNA
to
determine
whether
knockdown of any of the potential targets was able to phenocopy
treatment with the drug (i.e. to selectively affect viability of K-RAS
mutant cells) (Fig. S1c, Table S2). For these studies, we utilized the
DLD-1/DKs-8 isogenic cell lines because we had used this pair for
genetic analysis of SYK (Fig. 2). Knockdown of only one of the
potential targets that we assayed, HIPK2, was able to selectively
affect viability of K-RAS mutant cells (Fig. 3b). Nevertheless, we PLoS ONE | www.plosone.org July 2012 | Volume 7 | Issue 7 | e41343 4 Selective Activity of BAY61-3606 Figure 3. Identification and attempted validation of BAY61-3606 targets. (a) TREEspot image representing the inhibitory activity of BAY61-
3606. Kinases that are inhibited for ATP binding by BAY61-3606 are indicated by red dots on the phylogenetic tree of kinases. The size of the red dot
corresponds to the amount of inhibition by 1 mM BAY61-3606. The identity of the individual kinases can be found in Table S1. (b) Cell viability
quantified by Syto60 after shRNA-mediated knockdown of the potential BAY61-3606 targets in DLD-1 (red) or DKs-8 (blue) cell lines. Relative cell
viability is normalized to the parental cell lines infected with vector only. Error bars represent SEM for 2 independent experiments. doi:10.1371/journal.pone.0041343.g003 Figure 3. Identification and attempted validation of BAY61-3606 targets. (a) TREEspot image representing the inhibitory activity of BAY61-
3606. Kinases that are inhibited for ATP binding by BAY61-3606 are indicated by red dots on the phylogenetic tree of kinases. The size of the red dot
corresponds to the amount of inhibition by 1 mM BAY61-3606. The identity of the individual kinases can be found in Table S1. (b) Cell viability
quantified by Syto60 after shRNA-mediated knockdown of the potential BAY61-3606 targets in DLD-1 (red) or DKs-8 (blue) cell lines. Relative cell
viability is normalized to the parental cell lines infected with vector only. Error bars represent SEM for 2 independent experiments. doi:10.1371/journal.pone.0041343.g003 BAY61-3606 alters the response of cell expressing wild-type K-
RAS to AZ-628 by inhibiting the kinase activity of MAP4K2. First, we found that the basal level of Ikba phosphorylation was
significantly lower in HCT-116 than in HKe-3 cells, suggesting
that NFkb signaling is suppressed in cells expressing activated K-
RAS (Fig. S7). enetic analysis of potential BAY61-3606 targets Ikba phosphorylation was further induced in HKe-
3 cells after treatment with AZ-628 and this induction was
dependent upon MAP4K2 (Fig. 6a). This observation is consistent
with previous studies linking MAP4K2 to NFkb signaling [21]. Second, we found that JNK was strongly activated in HKe-3 cells
after treatment with AZ-628 (Fig. S7). Although MAP4K2 has
been previously linked to JNK signaling, this activation did not
appear to require MAP4K2 (Fig. S6b). MAP4K2 activates the NFkB pathway to counteract
AZ628-dependent growth inhibition Given that MAP4K2 is important for the response of wild-type
cells to AZ-628, we next asked how MAP4K2 functions to regulate
the response to RAF inhibition. To identify the pathway
responsible for MAP4K2 action, we used Bio-Plex phospho-
protein analysis to measure the effects of MAP4K2 knockdown on
various cellular signaling pathways. In total, we profiled 13
phospho-proteins: Ikba (Ser32/Ser36), JNK (Thr183/Tyr185),
MEK1 (Ser217/Ser221), ERK1/2 (Thr202/Tyr204, Thr185/
Tyr187), RSK (Thr359/Ser363), p38 (Thr180/Tyr182), c-JUN
(Ser63), ATF2 (Thr71), AKT (Ser473), S6 (Ser235/Ser236),
STAT3 (Ser727), STAT3 (Tyr705), and GSK3a/b (Ser21/Ser9)
(Fig. S7). We found two differences between HKe-3 and HCT-
116 that we posited might be related to the function of MAP4K2. Since NFkb was appeared to hyper-activated in wild-type cells
in a MAP4K2-dependent manner, we surmised that inhibition of
the NFkb pathway would have the same effect as BAY61-3606 or
MAP4K2 knockdown. That is, we expected that inhibition of
NFkb would increase the sensitivity of K-RAS wild-type cells to
AZ-628 without affecting the sensitivity of K-RAS mutant cells. As
predicted, inhibition of NFkb increased the sensitivity of HKe-3
and Dks-8 cells to AZ-628, essentially phenocopying MAP4K2
knockdown (Fig. 6b, Fig. S6c). By contrast, inhibition of JNK did
not alter the sensitivity of HKe-3 cells to AZ-628 (Fig. S6d). Further, as with BAY61-3606 treatment, knockdown of MAP4K2
abolished the ability of NFkb inhibition to sensitize wild-type cells
to AZ-628 (Fig. 6c). From these data, we conclude that MAP4K2
functions upstream in the NFkb pathway to regulate the response
of colorectal cancer cells to inhibition of RAF. Table 1. Activity of BAY61-3606 against selected kinases. Kinase
% inhibition
ATP binding*
IC50 (nM)**
CDK9
44
34.8
GSK3a
83
89.2
GSK3b
61
123
HIPK1
91.2
4240
HIPK2
91
248
MAP4K2
94
11.3
NEK1
73
159
ROCK1
96.8
144
ROCK2
87
166
SYK
93.9
25.5
*% inhibition of ATP binding as assessed by KINOMEscanTM (Ambit Biosciences). **In vitro IC50 were determined by SelectScreenH (Invitrogen). doi:10.1371/journal.pone.0041343.t001 Table 1. Activity of BAY61-3606 against selected kinases. Discussion K-RAS is mutationally activated in approximately 40% of
colorectal cancers [2]. Activated K-RAS is thought to confer
oncogenicity via its canonical downstream signaling pathways, for
example the RAF-MEK-ERK (MAPK) signaling cascade. Con-
sistent with this idea, activating mutations in B-RAF occur in 15%
of colorectal cancers and they are mutually exclusive with K-RAS
mutations [22]. Nevertheless, inhibition of MAPK signaling,
typically by directly inhibiting MEK, has been largely ineffective
in treating K-RAS mutant colorectal cancer [9,23]. The lack of
efficacy of MEK inhibitors in this context may be due to the
pleiotropic function of K-RAS, which has been shown to promote *% inhibition of ATP binding as assessed by KINOMEscanTM (Ambit Biosciences). **In vitro IC50 were determined by SelectScreenH (Invitrogen). doi:10.1371/journal.pone.0041343.t001 July 2012 | Volume 7 | Issue 7 | e41343 July 2012 | Volume 7 | Issue 7 | e41343 PLoS ONE | www.plosone.org 5 Selective Activity of BAY61-3606 Figure 4. Biological activities of BAY61-3606 derivatives. (a) GI50 values for BAY61-3606 derivatives performed in HCT-116 (red) or HKe-3
(blue) cells. Derivative 6 was chosen for further study for its increased potency and specificity for K-RAS mutant cells. (b) Cell viability quantified by
Syto60 after 72 hours exposure to BAY derivative 6 in HCT-116 (red) or HKe-3 (blue) cells. Relative cell viability was normalized to DMSO vehicle
treated control for each cell line. Error bars represent SEM for 3 independent experiments. HCT-116 cells were relatively sensitive to BAY derivative 6. doi:10.1371/journal.pone.0041343.g004 Figure 4. Biological activities of BAY61-3606 derivatives. (a) GI50 values for BAY61-3606 derivatives performed in HCT-116 (red) or HKe-3
(blue) cells. Derivative 6 was chosen for further study for its increased potency and specificity for K-RAS mutant cells. (b) Cell viability quantified by
Syto60 after 72 hours exposure to BAY derivative 6 in HCT-116 (red) or HKe-3 (blue) cells. Relative cell viability was normalized to DMSO vehicle
treated control for each cell line. Error bars represent SEM for 3 independent experiments. HCT-116 cells were relatively sensitive to BAY derivative 6. doi:10.1371/journal.pone.0041343.g004 Figure 5. MAP4K2 is a target for BAY61-3606 that modulates the response of wild-type cells to AZ-628. (a) Cell viability quantified by
Syto60 after 72 hours of combinatorial treatment with varying concentrations of BAY61-3606 and 1 mM AZ-628 in HCT-116 (red line) or HKe-3 (blue
lines) cells. Relative cell viability was normalized to DMSO vehicle treated control for each cell line. Discussion Error bars represent SEM for 3 independent
experiments. The two inhibitors cooperate in wild-type cells, but not in cells expressing mutant K-RAS. (b) Cell viability after 72 hours of
combinatorial treatment with varying concentrations of BAY derivative 6 and 1 mM AZ-628 in HCT-116 and HKe-3 cells. BAY derivative 6 does not
confer additional sensitivity to AZ-628 upon HKe-3 cells. (c) Cell viability quantified by Syto60 after 72 hours of AZ-628 treatment in HCT-116 or HKe-3
cell lines with MAP4K2 knockdown. Loss of MAP4K2 does not affect AZ-628 response in cells expressing mutant K-RAS, but enhances the effect of AZ-
628 in cells expressing wild-type K-RAS. (d) Cell viability after 72 hours of combinatorial treatment with 1 mM BAY61-3606 and 1 mM AZ-628 (shaded)
or 1 mM AZ-628 alone (clear) in HKe-3 cells with MAP4K2 knockdown. Relative cell viability was normalized to 1 mM AZ-628 treated samples. In
parental HKe-3 cells, BAY61-3606 confers sensitivity to AZ-628. Upon loss of MAP4K2, BAY61-3606 no longer sensitizes. doi:10.1371/journal.pone.0041343.g005 Figure 5. MAP4K2 is a target for BAY61-3606 that modulates the response of wild-type cells to AZ-628. (a) Cell viability quantified by
Syto60 after 72 hours of combinatorial treatment with varying concentrations of BAY61-3606 and 1 mM AZ-628 in HCT-116 (red line) or HKe-3 (blue
lines) cells. Relative cell viability was normalized to DMSO vehicle treated control for each cell line. Error bars represent SEM for 3 independent
experiments. The two inhibitors cooperate in wild-type cells, but not in cells expressing mutant K-RAS. (b) Cell viability after 72 hours of
combinatorial treatment with varying concentrations of BAY derivative 6 and 1 mM AZ-628 in HCT-116 and HKe-3 cells. BAY derivative 6 does not
confer additional sensitivity to AZ-628 upon HKe-3 cells. (c) Cell viability quantified by Syto60 after 72 hours of AZ-628 treatment in HCT-116 or HKe-3
cell lines with MAP4K2 knockdown. Loss of MAP4K2 does not affect AZ-628 response in cells expressing mutant K-RAS, but enhances the effect of AZ-
628 in cells expressing wild-type K-RAS. (d) Cell viability after 72 hours of combinatorial treatment with 1 mM BAY61-3606 and 1 mM AZ-628 (shaded)
or 1 mM AZ-628 alone (clear) in HKe-3 cells with MAP4K2 knockdown. Relative cell viability was normalized to 1 mM AZ-628 treated samples. In
parental HKe-3 cells, BAY61-3606 confers sensitivity to AZ-628. Upon loss of MAP4K2, BAY61-3606 no longer sensitizes. Discussion doi:10.1371/journal.pone.0041343.g005 July 2012 | Volume 7 | Issue 7 | e41343 PLoS ONE | www.plosone.org 6 Selective Activity of BAY61-3606 Figure 6. MAP4K2 modulates NFkb signaling. (a) Time course of phospho-Ikba (Ser32/Ser36) activation after 1 mM AZ-628 treatment in HCT-
116 (red lines) or HKe-3 (blue line) cells with MAP4K2 knock down, as measured via Bio-Plex. Relative signal was normalized to a master control lysate. Error bars represent SEM for 3 independent experiments. NFkb signaling was enhanced in HKe-3 cells after exposure to AZ-628 and this was
dependent upon MAP4K2. (b) Cell viability quantified by Syto60 after 72 hours of combinatorial treatment with IKK inhibitor VII and 1 mM AZ-628. Relative cell viability was normalized to DMSO vehicle treated control for each cell line. Like BAY61-3606, IKK inhibitor VII enhanced the effect of AZ-
628 specifically in K-RAS wild-type cells. (c) Cell viability after 72 hours of combinatorial treatment of 1 mM IKK inhibitor VII and 1 mM AZ-628 (shaded)
or 1 mM AZ-628 alone (clear) in HKe3 cells with MAP4K2 knockdown. Relative cell viability was normalized to 1 mM AZ-628 treated samples. Loss of
MAP4K2 abrogated the ability of IKK inhibitor VII to sensitize HKe-3 cells to AZ-628. doi:10.1371/journal.pone.0041343.g006 Figure 6. MAP4K2 modulates NFkb signaling. (a) Time course of phospho-Ikba (Ser32/Ser36) activation after 1 mM AZ-628 treatment in HCT-
116 (red lines) or HKe-3 (blue line) cells with MAP4K2 knock down, as measured via Bio-Plex. Relative signal was normalized to a master control lysate. Error bars represent SEM for 3 independent experiments. NFkb signaling was enhanced in HKe-3 cells after exposure to AZ-628 and this was
dependent upon MAP4K2. (b) Cell viability quantified by Syto60 after 72 hours of combinatorial treatment with IKK inhibitor VII and 1 mM AZ-628. Relative cell viability was normalized to DMSO vehicle treated control for each cell line. Like BAY61-3606, IKK inhibitor VII enhanced the effect of AZ-
628 specifically in K-RAS wild-type cells. (c) Cell viability after 72 hours of combinatorial treatment of 1 mM IKK inhibitor VII and 1 mM AZ-628 (shaded)
or 1 mM AZ-628 alone (clear) in HKe3 cells with MAP4K2 knockdown. Relative cell viability was normalized to 1 mM AZ-628 treated samples. Loss of
MAP4K2 abrogated the ability of IKK inhibitor VII to sensitize HKe-3 cells to AZ-628. Figure 6. MAP4K2 modulates NFkb signaling. Discussion (a) Time course of phospho-Ikba (Ser32/Ser36) activation after 1 mM AZ-628 treatment in HCT-
116 (red lines) or HKe-3 (blue line) cells with MAP4K2 knock down, as measured via Bio-Plex. Relative signal was normalized to a master control lysate. Error bars represent SEM for 3 independent experiments. NFkb signaling was enhanced in HKe-3 cells after exposure to AZ-628 and this was
dependent upon MAP4K2. (b) Cell viability quantified by Syto60 after 72 hours of combinatorial treatment with IKK inhibitor VII and 1 mM AZ-628. Relative cell viability was normalized to DMSO vehicle treated control for each cell line. Like BAY61-3606, IKK inhibitor VII enhanced the effect of AZ-
628 specifically in K-RAS wild-type cells. (c) Cell viability after 72 hours of combinatorial treatment of 1 mM IKK inhibitor VII and 1 mM AZ-628 (shaded)
or 1 mM AZ-628 alone (clear) in HKe3 cells with MAP4K2 knockdown. Relative cell viability was normalized to 1 mM AZ-628 treated samples. Loss of
MAP4K2 abrogated the ability of IKK inhibitor VII to sensitize HKe-3 cells to AZ-628. doi:10.1371/journal.pone.0041343.g006 its biologically active derivatives is a non-kinase protein. Aside
from inhibiting kinases, ATP analogs can affect biology through
other processes, including nucleic acids synthesis [29,30] and
microtubule motor transport [31]. Alternately, the relevant target
may not be among the 402 kinases that were surveyed in our assay
or BAY61-3606 and derivatives could inhibit kinase activity
without affecting active site binding. If so, they would not score in
our screen. transformation through the PI3K and RAL effector pathways in
addition to MAPK [24,25]. Yet, PI3K pathway mutations also
occur in colorectal cancers and are often coincident with K-RAS
mutations, suggesting that PI3K is not a common effector of K-
RAS signaling in colon cancer [26]. And while PI3K mutations
have been associated with resistance to MEK inhibitors in cancer
cell lines [27,28], K-RAS mutant colon cancers from genetically
engineered mice are intrinsically resistant to inhibition of MEK
[9]. Our data are consistent with an alternative explanation for the
lack of efficacy of MEK inhibitors in colorectal cancers expressing
mutant K-RAS – that there exists an alternate/parallel pathway
downstream of B-RAF that mediates K-RAS-induced oncogenic-
ity. In addition to its activity in cells expressing mutant K-RAS or
B-RAF, we also identified a secondary biological effect of BAY61-
3606; it conferred upon wild-type cells, which are normally
resistant to AZ-628, sensitivity to RAF inhibition (Fig. 5a). Discussion Using
a variety of approaches, we identified MAP4K2 as the target for
BAY61-3606 in wild-type cells. MAP4K2 (also known as GCK,
for Germinal Center Kinase) is a member of the STE20 family of
protein kinases [32]. MAP4K2 has been shown to play a role in
the inflammatory process and is activated by inflammatory stimuli
such as TNFa, IL-1, and LPS [33,34]. MAP4K2 interacts with
TNFR-associated factor 2 (TRAF2) and MAPK/ERK kinase
kinase 1 (MEKK1), thereby linking TNF signaling to activation of
the pro-death JNK and p38 MAPK pathways [35]. Aside from
activating pro-death signaling pathways, the TNF receptor
pathway can also promote cell survival through the NFkb
pathway. Incidentally, the NFkb pathway is recruited by the
TRAF2
complex
upon
TNFR
activation
[36]. Moreover,
MAP4K2 has been shown to positively regulate NFkb to protect
melanoma cells from UV-induced apoptosis [21]. In our study, we characterized the activity of a small molecule,
BAY61-3606, that preferentially affected viability in colorectal
cancer cells expressing mutant K-RAS compared to isogenic cells
expressing only wild-type K-RAS (Fig. 1a, Fig. S1a). Since
BAY61-3606 is an ATP-competitive kinase inhibitor, its ability
to preferentially affect cells expressing mutant K-RAS initially
suggested that it targets a kinase functioning downstream of K-
RAS to promote proliferation. We have previously shown that K-
RAS promotes colon cancer cell proliferation through a RAF-
dependent, but MEK-independent, signaling pathway [9]. Three
pieces of evidence implicate BAY61-3606 as an inhibitor of this
MEK-independent pathway downstream of RAF. First, BAY61-
3606 did not cooperate with AZ-628 in cells expressing mutant K-
RAS (Fig. 5a), suggesting that these inhibitors targeted a common
pathway. Second, BAY61-3606 affected growth in KRAS mutant
cells, but, unlike AZ-628, did not affect the phosphorylation state
of MEK or ERK (Fig. 1d). Finally, BAY61-3606 slowed the
growth of colorectal cancer cells expressing mutant B-RAF and
cooperated with a MEK inhibitor to produce an enhanced
response in these cells (Fig. 1c). Inhibition of RAF by AZ-628 leads to MAP4K2-dependent
activation of NFkb signaling in wild-type cells, presumably to
ensure cell survival (Fig. 6a). To maintain reliable behavior, cell
signaling networks have evolved robust feedback mechanisms in
order to minimize the effects of focal perturbations, which can
arise from various stresses that cells encounter. The robustness of
a signaling network is evident in its compensatory behavior given
the activation or inactivation of a single pathway. PLoS ONE | www.plosone.org Table S1
Active site binding inhibition data.
(PDF) Table S2
shRNAs used in this study. (PDF) Discussion For example, it
has been demonstrated that RAF pathway activity is anti-
correlated to PI3K signaling through global network feedback
[37]. Consequently, pharmacologic inhibition of MEK results in
PI3K pathway activation, resulting in cell survival [38]. Similarly,
inhibition of mTOR, a downstream effector of PI3K, has been Although BAY61-3606 was initially characterized as an ATP
competitive kinase inhibitor, its selectivity for decreasing the
viability of K-RAS mutant cells may not require this activity. Our
studies of BAY61-3606 derivatives demonstrate that those lacking
the ability to affect active site binding can still maintain their
ability to selectively affect cell viability. One possible explanation
for this observation is that the relevant target of BAY61-3606 and PLoS ONE | www.plosone.org July 2012 | Volume 7 | Issue 7 | e41343 7 Selective Activity of BAY61-3606 after 72 hours of BAY derivative 6 treatment in 5 cell lines
expressing mutant K-RAS and in 3 cell lines expressing wild-type
K-RAS. Relative cell viability was normalized to an untreated
control for each cell line. Error bars represent SEM for 3
independent experiments. With the exception of GP5d (highlight
in bold red), all of the cell lines expressing mutant K-RAS respond
to BAY derivative 6. (PDF) shown to increase ERK activity [39]. Given the large number of
mutations that cancer cells accumulate, oncogenic signaling
networks have evolved to be very different from signaling networks
in normal cells. Whereas cancer cells have evolved such that they
are robust to growth and proliferation, there may be areas in their
networks that are more fragile to perturbations than their wild-
type counterparts, which can be exploited for therapeutic
purposes. While our studies specifically address the function of the small
molecule kinase inhibitor BAY61-3606 in colorectal cancer cells,
they more generally address the varied activities that kinase
inhibitors can have in cancer cells. Although inhibitor promiscuity
is often viewed in a negative light, the ability of a given small
molecule to target multiple kinases may, in fact, be beneficial for
targeting diverse genotypic classes of cancer. Figure S4
BAY61-3606 derivatives lose ATP competitive
activity. TREEspot images for five different derivatives of
BAY61-3606. Both inhibitors that retained selectivity for K-RAS
mutant cells (e.g. 6 and 8) and those that lost selectivity (e.g. 1, 21,
and 28), failed to effectively inhibit ATP binding by the majority of
kinases that were assayed. Supporting Information Figure S5
Kinase inhibition profiles of BAY61-3606 and
its derivatives. Inhibitor activity was measured using Invitro-
gen’s SelectScreenH Biochemical Kinase Profiling Service. (PDF) Figure S1
Genetic analysis of BAY61-3606 response in
DLD-1 cells. (a) Cell viability quantified by Syto60 after
72 hours of BAY61-3606 treatment in DLD-1 (K-RASG13D/+,
red) or DKs-8 (K-RAS2/+, blue) cell lines. Relative cell viability
was normalized to DMSO vehicle treated control for each cell
line. Error bars represent SEM for 3 independent experiments. The differential response in the two cell lines is statistically
significant (p = 0.019 at 1 mM). (b) Cell viability quantified by
Syto60 after 72 hours of BAY derivative 6 treatment in DLD-1
(K-RASG13D/+, red) or DKs-8 (K-RAS2/+, blue) cell lines. Relative cell viability was normalized to DMSO vehicle treated
control for each cell line. Error bars represent SEM for 3
independent experiments. (c) Validation of shRNAs. Relative gene
expression of shRNA-mediated knockdowns of potential BAY61-
3606 targets in DLD-1 (red) and DKs-8 (blue) cells. Gene
expression is measured via Taqman assay and calculated using
standard methods in reference to the housekeeping gene TBP. Error bars represent SEM for 3 independent experiments. (PDF) Figure S6
Evaluation of MAP4K2 in the BAY61-3606
response. (a) Cell viability quantified by Syto60 after 72 hours of
AZ-628 treatment in DLD-1 or DKs-8 cell lines with MAP4K2
knockdown. Loss of MAP4K2 does not affect AZ-628 response in
cells expressing mutant K-RAS, but enhances the effect of AZ-628
in cells expressing wild-type K-RAS. (b) Time course of phospho-
JNK (Thr183/Tyr185) after 1 mM AZ-628 treatment in HCT-116
(red lines) or HKe-3 (blue line) cells with MAP4K2 knock down, as
measured by Bio-Plex. Relative signal was normalized to a master
control lysate. Error bars represent SEM for 3 independent
experiments. JNK signaling was enhanced in HKe-3 cells but was
independent of MAP4K2. (c) Cell viability quantified by Syto60
after 72 hours of combinatorial treatment with IKK inhibitor VII
and 1 mM AZ-628. Relative cell viability was normalized to
DMSO vehicle treated control for each cell line. Like BAY61-
3606, IKK
inhibitor VII
enhanced
the
effect
of AZ-628
specifically in K-RAS wild-type cells. (d) Cell viability quantified
by Syto60 after 72 hours of combinatorial treatment with the JNK
inhibitor SP600125 and 1 mM AZ-628. Relative cell viability was
normalized to DMSO vehicle treated control for each cell line. Unlike BAY61-3606, SP600125 did not affect AZ-628 sensitivity
in K-RAS wild-type cells. (PDF) Figure S7
Signaling pathway activity in response to AZ-
628. Phospho-protein measurements were made after 45 minutes
of exposure to 1 mM AZ-628 or DMSO vehicle control treatment
in HCT-116 (red squares) or HKe-3 (blue dots) cells. X’s represent
measurements
from
cells with MAP4K2
knock down. All
measurements were quantified by Bio-Plex signaling assays. Relative signal was normalized to a master control lysate. (PDF) Table S1
Active site binding inhibition data. (PDF) Supporting Information (PDF) Figure S2
Chemical derivation of BAY61-3606 deriva-
tives. (a) Synthesis of BAY derivative 6. To a stirred solution of
5,7-dichloroimidazo [1,5-f] pyrimidine (186.0 mg, 1.0 mmol) in
DMF (5.0 mL) was added 2-methoxybenzenamine (123.0 mg, 1.0
equiv). After 1 h heating at 70u, the mixture was purified on silica
gel column with methylene chloride and methanol (10:1) as eluent
to give of 7-chloro-N-(2-methoxyphenyl)imidazo [1,5-f] pyrimidin-
5-amine(245 mg, yield 89%). To a solution of 7-chloro-N-(2-
methoxyphenyl)imidazo
[1,5-f]
pyrimidin-5-amine
(200.0 mg,
0.73 mmol) and 3,4-dimethoxyphenylboronic acid (160.0 mg,
1.2 equiv) in 5.0 mL 1,4-dioxane was added Bis(triphenylpho-
sphine) palladium(II) dichloride (51.0 mg, 0.1 equiv) as catalyst
and saturated potassium carbonate aqueous solution (2.0 mL) as
base. The mixture was heated for 2 h at 80u and then was diluted
with chloroform and 2-propanol (50 mL, 4:1). The organic layer
was washed with water, brine and was dried with sodium sulfate. After removal of solvent, the crude was purified by column with
methylene chloride and methanol (10:1) to give BAY derivative 6
(192.0 mg, 70%). 1H NMR (DMSO-d6) 9.05 (s, 1 H), 8.22 (s, 1 H),
7.23 (d, J = 7.8 Hz, 1 H), 7,61-7.53 (m, 4 H), 7.26 (t, J = 7.8 Hz, 1
H), 7.16 (d, J = 8.4 Hz, 1 H), 7.04 (t, J = 7.8 Hz, 1 H), 6.94 (d,
J = 8.4 Hz, 1 H), 3.80 (s, 3 H), 3.75 (s, 3 H), 3.72 (s, 3 H). (b)
Chemical structures of all BAY derivatives. (PDF) Figure S2
Chemical derivation of BAY61-3606 deriva-
tives. (a) Synthesis of BAY derivative 6. To a stirred solution of
5,7-dichloroimidazo [1,5-f] pyrimidine (186.0 mg, 1.0 mmol) in
DMF (5.0 mL) was added 2-methoxybenzenamine (123.0 mg, 1.0
equiv). After 1 h heating at 70u, the mixture was purified on silica
gel column with methylene chloride and methanol (10:1) as eluent
to give of 7-chloro-N-(2-methoxyphenyl)imidazo [1,5-f] pyrimidin-
5-amine(245 mg, yield 89%). To a solution of 7-chloro-N-(2-
methoxyphenyl)imidazo
[1,5-f]
pyrimidin-5-amine
(200.0 mg,
0.73 mmol) and 3,4-dimethoxyphenylboronic acid (160.0 mg,
1.2 equiv) in 5.0 mL 1,4-dioxane was added Bis(triphenylpho-
sphine) palladium(II) dichloride (51.0 mg, 0.1 equiv) as catalyst
and saturated potassium carbonate aqueous solution (2.0 mL) as
base. The mixture was heated for 2 h at 80u and then was diluted
with chloroform and 2-propanol (50 mL, 4:1). The organic layer
was washed with water, brine and was dried with sodium sulfate. Supporting Information After removal of solvent, the crude was purified by column with
methylene chloride and methanol (10:1) to give BAY derivative 6
(192.0 mg, 70%). 1H NMR (DMSO-d6) 9.05 (s, 1 H), 8.22 (s, 1 H),
7.23 (d, J = 7.8 Hz, 1 H), 7,61-7.53 (m, 4 H), 7.26 (t, J = 7.8 Hz, 1
H), 7.16 (d, J = 8.4 Hz, 1 H), 7.04 (t, J = 7.8 Hz, 1 H), 6.94 (d,
J = 8.4 Hz, 1 H), 3.80 (s, 3 H), 3.75 (s, 3 H), 3.72 (s, 3 H). (b)
Chemical structures of all BAY derivatives. (PDF) References Cancer Cell 18: 63–73. 33. Pombo CM, Kehrl JH, Sanchez I, Katz P, Avruch J, et al. (1995) Activation of
the SAPK pathway by the human STE20 homologue germinal centre kinase. Nature 377: 750–754. 13. Luo J, Emanuele MJ, Li D, Creighton CJ, Schlabach MR, et al. (2009) A
genome-wide RNAi screen identifies multiple synthetic lethal interactions with
the Ras oncogene. Cell 137: 835–848. 34. Zhong J, Gavrilescu LC, Molnar A, Murray L, Garafalo S, et al. (2009) GCK is
essential to systemic inflammation and pattern recognition receptor signaling to
JNK and p38. Proc Natl Acad Sci U S A 106: 4372–4377. g
14. Yamamoto N, Takeshita K, Shichijo M, Kokubo T, Sato M, et al. (2003) The
orally available spleen tyrosine kinase inhibitor 2-[7-(3,4-dimethoxyphenyl)-
imidazo [1,2-c] pyrimidin-5-ylamino] nicotinamide dihydrochloride (BAY 61-
3606) blocks antigen-induced airway inflammation in rodents. J Pharmacol Exp
Ther 306: 1174–1181. 35. Yuasa T, Ohno S, Kehrl JH, Kyriakis JM (1998) Tumor necrosis factor signaling
to stress-activated protein kinase (SAPK)/Jun NH2-terminal kinase (JNK) and
p38. Germinal center kinase couples TRAF2 to mitogen-activated protein
kinase/ERK kinase kinase 1 and SAPK while receptor interacting protein
associates with a mitogen-activated protein kinase kinase kinase upstream of
MKK6 and p38. J Biol Chem 273: 22681–22692. 15. Pearce G, Audzevich T, Jessberger R (2011) SYK regulates B-cell migration by
phosphorylation of the F-actin interacting protein SWAP-70. Blood 117: 1574–
1584. 36. Natoli G, Costanzo A, Guido F, Moretti F, Bernardo A, et al. (1998) Nuclear
factor kB-independent cytoprotective pathways originating at tumor necrosis
factor receptor-associated factor 2. J Biol Chem 273: 31262–31272. 16. Ventura A, Meissner A, Dillon CP, McManus M, Sharp PA, et al. (2004) Cre-
lox-regulated conditional RNA interference from transgenes. Proc Natl Acad
Sci U S A 101: 10380–10385. 37. Courtois-Cox S, Genther Williams SM, Reczek EE, Johnson BW, McGillicuddy
LT, et al. (2006) A negative feedback signaling network underlies oncogene-
induced senescence. Cancer Cell 10: 459–472. 17. Shirasawa S, Furuse M, Yokoyama N, Sasazuki T (1993) Altered growth of
human colon cancer cell lines disrupted at activated Ki-ras. Science 260: 85–88. 18. Montagut C, Sharma SV, Shioda T, McDermott U, Ulman M, et al. (2008)
Elevated CRAF as a potential mechanism of acquired resistance to BRAF
inhibition in melanoma. Cancer Res 68: 4853–4861. 38. Gopal YN, Deng W, Woodman SE, Komurov K, Ram P, et al. References Sebolt-Leopold JS, Dudley DT, Herrera R, Van Becelaere K, Wiland A, et al. (1999) Blockade of the MAP kinase pathway suppresses growth of colon tumors
in vivo. Nat Med 5: 810–816. 27. Balmanno K, Chell SD, Gillings AS, Hayat S, Cook SJ (2009) Intrinsic
resistance to the MEK1/2 inhibitor AZD6244 (ARRY-142886) is associated
with weak ERK1/2 signalling and/or strong PI3K signalling in colorectal cancer
cell lines. International journal of cancer Journal international du cancer 125:
2332–2341. 7. Solit DB, Garraway LA, Pratilas CA, Sawai A, Getz G, et al. (2006) BRAF
mutation predicts sensitivity to MEK inhibition. Nature 439: 358–362. 8. Jhawer M, Goel S, Wilson AJ, Montagna C, Ling YH, et al. (2008) PIK3CA
mutation/PTEN expression status predicts response of colon cancer cells to the
epidermal growth factor receptor inhibitor cetuximab. Cancer Res 68: 1953–
1961. 28. Wee S, Jagani Z, Xiang KX, Loo A, Dorsch M, et al. (2009) PI3K pathway
activation mediates resistance to MEK inhibitors in KRAS mutant cancers. Cancer Res 69: 4286–4293. 29. Saikrishnan K, Powell B, Cook NJ, Webb MR, Wigley DB (2009) Mechanistic
basis of 59–39 translocation in SF1B helicases. Cell 137: 849–859. 9. Haigis KM, Kendall KR, Wang Y, Cheung A, Haigis MC, et al. (2008)
Differential effects of oncogenic K-Ras and N-Ras on proliferation, differenti-
ation and tumor progression in the colon. Nat Genet 40: 600–608. 30. Liu Y, Kung C, Fishburn J, Ansari AZ, Shokat KM, et al. (2004) Two cyclin-
dependent kinases promote RNA polymerase II transcription and formation of
the scaffold complex. Mol Cell Biol 24: 1721–1735. 10. Barbie DA, Tamayo P, Boehm JS, Kim SY, Moody SE, et al. (2009) Systematic
RNA interference reveals that oncogenic KRAS-driven cancers require TBK1. Nature 462: 108–112. 31. Zhao YC, Kull FJ, Cochran JC (2010) Modulation of the kinesin ATPase cycle
by neck linker docking and microtubule binding. J Biol Chem 285: 25213–
25220. 11. Scholl C, Frohling S, Dunn IF, Schinzel AC, Barbie DA, et al. (2009) Synthetic
lethal interaction between oncogenic KRAS dependency and STK33 suppres-
sion in human cancer cells. Cell 137: 821–834. 32. Dan I, Watanabe NM, Kusumi A (2001) The Ste20 group kinases as regulators
of MAP kinase cascades. Trends Cell Biol 11: 220–230. 12. Puyol M, Martin A, Dubus P, Mulero F, Pizcueta P, et al. (2010) A synthetic
lethal interaction between K-Ras oncogenes and Cdk4 unveils a therapeutic
strategy for non-small cell lung carcinoma. References 21. Ivanov VN, Kehrl JH, Ronai Z (2000) Role of TRAF2/GCK in melanoma
sensitivity to UV-induced apoptosis. Oncogene 19: 933–942. 1. Malumbres M, Barbacid M (2003) RAS oncogenes: the first 30 years. Nat Rev
Cancer 3: 459–465. 2. Lau KS, Haigis KM (2009) Non-redundancy within the RAS oncogene family:
Insights into mutational disparities in cancer. Mol Cells. 22. Davies H, Bignell GR, Cox C, Stephens P, Edkins S, et al. 22. Davies H, Bignell GR, Cox C, Stephens P, Edkins S, et al. (2002) Mutations of
the BRAF gene in human cancer. Nature 417: 949–954. the BRAF gene in human cancer. Nature 417: 949–954. Insights into mutational disparities in cancer. Mol Cells. 3. Benvenuti S, Sartore-Bianchi A, Di Nicolantonio F, Zanon C, Moroni M, et al. (2007) Oncogenic activation of the RAS/RAF signaling pathway impairs the
response of metastatic colorectal cancers to anti-epidermal growth factor
receptor antibody therapies. Cancer Res 67: 2643–2648. g
23. Wang Y, Van Becelaere K, Jiang P, Przybranowski S, Omer C, et al. (2005) A 23. Wang Y, Van Becelaere K, Jiang P, Przybranowski S, Omer C, et al. (2005) A
role for K-ras in conferring resistance to the MEK inhibitor, CI-1040. Neoplasia
7: 336–347. g
J
g
y
(
)
role for K-ras in conferring resistance to the MEK inhibitor, CI-1040. Neoplasia
7: 336–347. 24. Gupta S, Ramjaun AR, Haiko P, Wang Y, Warne PH, et al. (2007) Binding of
ras to phosphoinositide 3-kinase p110alpha is required for ras-driven
tumorigenesis in mice. Cell 129: 957–968. p
y
p
4. Di Fiore F, Blanchard F, Charbonnier F, Le Pessot F, Lamy A, et al. (2007)
Clinical relevance of KRAS mutation detection in metastatic colorectal cancer
treated by Cetuximab plus chemotherapy. Br J Cancer 96: 1166–1169. 25. Martin TD, Samuel JC, Routh ED, Der CJ, Yeh JJ (2011) Activation and
involvement of Ral GTPases in colorectal cancer. Cancer Res 71: 206–215. 5. De Roock W, Claes B, Bernasconi D, De Schutter J, Biesmans B, et al. (2010)
Effects of KRAS, BRAF, NRAS, and PIK3CA mutations on the efficacy of
cetuximab plus chemotherapy in chemotherapy-refractory metastatic colorectal
cancer: a retrospective consortium analysis. The lancet oncology 11: 753–762. 26. Velho S, Oliveira C, Ferreira A, Ferreira AC, Suriano G, et al. (2005) The
prevalence of PIK3CA mutations in gastric and colon cancer. European journal
of cancer 41: 1649–1654. 6. Acknowledgments We would like to thanks L. Kleiman and M. Patricelli for helpful comments
on the manuscript. We would like to thanks L. Kleiman and M. Patricelli for helpful comments
on the manuscript. Figure S3
Evaluation of BAY derivative 6 activity in
colorectal cancer cell lines. Cell viability quantified by Syto60 Figure S3
Evaluation of BAY derivative 6 activity in
colorectal cancer cell lines. Cell viability quantified by Syto60 July 2012 | Volume 7 | Issue 7 | e41343 PLoS ONE | www.plosone.org 8 Selective Activity of BAY61-3606 Author Contributions Conceived and designed the experiments: KSL DAL NSG KMH. Performed the experiments: KSL TZ KRK. Analyzed the data: KSL TZ
KRK DAL NSG KMH. Wrote the paper: KSL KMH. References (2010) Basal and
treatment-induced activation of AKT mediates resistance to cell death by
AZD6244 (ARRY-142886) in Braf-mutant human cutaneous melanoma cells. Cancer Res 70: 8736–8747. 19. Lyne PD, Aquila B, Cook DJ, Dakin LA, Ezhuthachan J, et al. (2009)
Identification of amidoheteroaryls as potent inhibitors of mutant (V600E) B-Raf
kinase with in vivo activity. Bioorg Med Chem Lett 19: 1026–1029. 39. Carracedo A, Ma L, Teruya-Feldstein J, Rojo F, Salmena L, et al. (2008)
Inhibition of mTORC1 leads to MAPK pathway activation through a PI3K-
dependent feedback loop in human cancer. J Clin Invest 118: 3065–3074. y
g
20. Karaman MW, Herrgard S, Treiber DK, Gallant P, Atteridge CE, et al. (2008)
A quantitative analysis of kinase inhibitor selectivity. Nat Biotechnol 26: 127–
132. PLoS ONE | www.plosone.org July 2012 | Volume 7 | Issue 7 | e41343 9
|
https://openalex.org/W4375848291
|
https://zenodo.org/records/7907184/files/Rapportage%20Sharehouse%205.3%20-%20Handvatten%20voor%20logistiek%20onderwijs%20v4%2012-04-2023.pdf
|
Dutch
| null |
D5.3-Incorporation of new required skills and teaching material in logistics curricula at STC, HR, Fontys and Windesheim during and immediately after the project.
|
Zenodo (CERN European Organization for Nuclear Research)
| 2,023
|
cc-by
| 16,412
|
2023 2023 2023 D5.3-Incorporation of new
required skills and teaching
material in logistics curricula at
STC, HR, Fontys and
Windesheim during and
immediately after the project. WELKE HANDVATTEN KUNNEN WORDEN
BENOEMD VOOR HET LOGISTIEKE ONDERWIJS,
ZOWEL MBO ALS HBO, OM KLAAR TE ZIJN VOOR
DE LOGISTIEKE VAARDIGHEDEN VAN DE
TOEKOMST?
DATUM: FEBRUARY 2023
AUTEURS:
GROLLEMAN, HENRI; HENDRICKX, RENÉ; WARFFEMIUS, PIM;
HEZEKEN, VAN ARJAN; VOS, MENNO
TYPE: PUBLIC 1
h
h
l b l
Dit onderzoeksproject maakt deel uit van het onderzoeksprogramma Duurzame Living Labs dat
(mede)gefinancierd is door de Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO),
Ministerie Infrastructuur & Waterstaat, Nationaal Regieorgaan Praktijkgericht Onderzoek (SIA) en de
Topsector Logistiek (Nummer 439.18.452 A). Inhoud
Inleiding ....................................................................................................................................................................... 3
HARD SKILLS ............................................................................................................................................................. 7
Analytische vaardigheden ............................................................................................................................ 7
Resultaten MBO-CuCo’s ........................................................................................................................... 7
Resultaten HBO-CuCo’s ............................................................................................................................ 8
Gaps & invulling ............................................................................................................................................. 9
Conclusie .......................................................................................................................................................... 9
Expert op vakgebied ...................................................................................................................................... 10
Resultaten MBO-CuCo’s ......................................................................................................................... 10
Resultaten HBO-CuCo’s ........................................................................................................................... 12
Conclusie ......................................................................................................................................................... 12
Voorbeelden van ‘eigen’ initiatieven ...................................................................................................... 13
Een toekomstbestendig warehouse .................................................................................................. 13
Deltion E-Logistics ..................................................................................................................................... 14
Experimenteren in een Living Lab: Exoskelets ............................................................................. 14
Datadriven Logistics ................................................................................................................................. 14
SOFT SKILLS ............................................................................................................................................................. 15
Resultaten CuCo’s MBO ................................................................................................................................ 15
Resultaten CuCo’s HBO ................................................................................................................................ 16
Conclusie ........................................................................................................................................................ 16
Voorbeelden van ‘eigen’ initiatieven ...................................................................................................... 17
Het Soft Skills Lab ........................................................................................................................................ 17
Deltion E-Logistics ......................................................................................................................................18
Leerlijn Professionele ontwikkeling ....................................................................................................18
De lerende organisatie ..............................................................................................................................18
ALGEMENE REFLECTIE ....................................................................................................................................... 19
ROL BEDRIJFSLEVEN ........................................................................................................................................... 19
Visie MBO ............................................................................................................................................................ 19
Visie HBO ............................................................................................................................................................. 19 1 Dit onderzoeksproject maakt deel uit van het onderzoeksprogramma Duurzame Living Labs dat
(mede)gefinancierd is door de Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO),
Ministerie Infrastructuur & Waterstaat, Nationaal Regieorgaan Praktijkgericht Onderzoek (SIA) en de
Topsector Logistiek (Nummer 439.18.452 A). Dit onderzoeksproject maakt deel uit van het onderzoeksprogramma Duurzame Living Labs dat
(mede)gefinancierd is door de Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO),
Ministerie Infrastructuur & Waterstaat, Nationaal Regieorgaan Praktijkgericht Onderzoek (SIA) en de
Topsector Logistiek (Nummer 439.18.452 A). Inhoud Rol van Living Lab ........................................................................................................................................... 20
LANDELIJK ONDERWIJS ................................................................................................................................... 20
MBO-onderwijs ................................................................................................................................................ 20
HBO-onderwijs .................................................................................................................................................. 21
HARD skills ...................................................................................................................................................... 22
SOFT skills ....................................................................................................................................................... 22
Conclusie ..................................................................................................................................................................23
Kritische reflectie ................................................................................................................................................ 25
Aanbevelingen voor Verder onderzoek ................................................................................................... 25
Bibliografie ............................................................................................................................................................... 27
Bijlage 01 – Resultatentabel Logistics Skills of the Future ( .......................................................... 28
Bijlage 02 – overzicht voorbeelden vanuit onderwijs ...................................................................... 29
Bijlage 03 – Data Science in het HBO-onderwijs (Fontys) ............................................................ 30
Bijlage 04 – Data Science in het HBO onderwijs (Hogeschool Rotterdam) .......................... 33
Bijlage 05 - Opzet Data Science for logistici (Hogeschool Rotterdam) ................................. 39
Bijlage 06 – Praktijkcasus STC ..................................................................................................................... 42
Bijlage 07 – Casus Deltion E-logistics ...................................................................................................... 43
Bijlage 08 – Lean Orange Belt ...................................................................................................................... 45
Bijlage 09 – De lerende organisatie ............................................................................................................ 47
Bijlage 10 - Professionele Ontwikkeling leerjaar 1 (Semester 1 & 2) .......................................... 49 www.sharehouselab.nl www.sharehouselab.nl 3 www.sharehouselab.nl Samenvatting Bij het inrichten van nieuwe en bestaande warehouses wordt in steeds grotere mate
digitalisering en robotisering toegepast. Diverse onderzoeken zijn reeds uitgevoerd
naar de impact op de banen van de toekomst in de logistiek door de digitalisering en
robotisering, hierbij wordt echter niet ingegaan op de veranderende skills van
medewerkers op verschillende opleidingsniveaus. In deze bijdrage worden de resultaten van gesprekken met verschillende Curricula
comissies van HBO en MBO opleidingen gedeeld. Vanuit dit onderzoek is gebleken dat
een aantal van de belangrijke veranderende skills worden ingevuld vanuit een
individuele opleiding, maar dat landelijke opleidingsprofielen hier nog niet geheel of in
voldoende mate bij aansluiten. Tevens wordt ervaren dat het ontwikkelen van
onderwijs steeds sneller plaats dient te vinden, waardoor het uitwisselen van onderwijs
belangrijker wordt. Hiertoe zijn in deze rapportage ook diverse voorbeelden van
onderwijs te vinden ter inspiratie. De belangrijkste resultaten kunnen worden gevonden in de oproep om meer
samenwerking te zoeken tussen onderwijs en bedrijfsleven. Het bedrijfsleven dient
meer geintegreerd te worden in het onderwijs. Daarnaast dienen onderwerpen als
datagedreven werken en digitalisering meer terug te komen op zowel HBO als MBO
onderwijs, waarbij ook de soft skills van de student een update nodig hebben. www.sharehouselab.nl 4 De toenemende digitalisering en robotisering in warehouses stelt nieuwe eisen aan
(toekomstige) warehouse medewerkers. Het rapport van Rathenau (Freese, Dekker,
Kool, Dekker, & Est, 2018) laat zien dat bijna elke baan van karakter verandert als gevolg
van robotisering en automatisering. Een belangrijke vraag is in hoeverre de logistieke
opleidingen toekomstbestendig zijn en goed aansluiten op de toekomstige behoeften
van werkgevers (Zijm & Klumpp, 2017). Daarvoor is het belangrijk om te weten welke
nieuwe skills (bestaande uit kennis, vaardigheden en gedrag) in middelbaar en hoger
beroepsonderwijs nodig zijn voor de logistieke sector en om eventuele mismatches
met het huidige curriculum inzichtelijk te maken. In het kader van het SHAREHOUSE
programma, waarin meer dan 20 partners samen onderzoek doen naar technische en
sociale innovatie in het magazijn van de toekomst (www.sharehouselab.nl), is een
Logistics Skills of the Future scan ontwikkeld (Grolleman, Hendrickx, Warffemius, Vos,
& Versluis, 2022). Een aantal van de skills die in dit onderzoek als ‘belangrijk’ zijn
benoemd, zijn het vertrekpunt geweest om met curriculum commissies (CuCo’s) en
opleidingsmanagers van HBO en MBO-opleidingen het gesprek aan te gaan. Het doel van deze rapportage is enerzijds een vergelijking te maken van een aantal
belangrijke skills vanuit de resultaten van de LSF-scan met Logistieke curicula op MBO
en HBO niveau. Anderzijds willen we recent ontwikkeld onderwijsaanbod op MBO en
HBO niveau delen die aansluiten op die verschuiving in gevraagde skills met als doel
logistieke opleidingen te inspireren en mogelijk zelfs aan te sporen tot samenwerking
aan vernieuwend’ onderwijsmaterieel. De onderzoeksvraag die ten grondslag aan deze rapportage heeft gelegen is als volgt:
“Welke skills kunnen we constateren in onderwijscurricula en op welke wijze dienen
kan deze worden gedicht met als doel; 1. Verhogen van bewustzijn van toekomstige
taken in de logistieke sector en 2. Verbeteren van aangeleerde skills voor toekomstige
taken in de logistieke sector.“ Er wordt gestart met een overzicht van de aanpak, waarin zowel de belangrijkste
resultaten vanuit de LSF-scan (Grolleman, Hendrickx, Warffemius, Vos, & Versluis,
2022) worden benoemd als ook een onderscheid wordt gemaakt naar het onderscheid
tussen de verschillende opleidingsniveaus. Vervolgens worden inzichten gegeven aan de hand van een indeling op hard en soft
skills inzicht gegeven in de resultaten aan de hand van gevoerde gesprekken. Dit wordt
continu opgebouwd aan de hand van een zo specifiek mogelijk gedefinieerde skill,
waarna inzicht wordt gegeven in de resultaten vanuit het MBO als HBO onderwijs. www.sharehouselab.nl www.sharehouselab.nl Na
een conclusie worden er per skill een aantal voorbeelden benoemd van aangeboden
onderwijs ter inspiratie. Na een reflectie op de resultaten, de rol en context van het bedrijfsleven in het
onderwijs wordt de vergelijking getrokken met de bevindingen met de huidige
(landelijke) opleidingsprofielen. Er wordt afgesloten met een conclusie waarin diverse
aanbevelingen worden gedaan met voor het logistieke onderwijs als ook enkele
aanbevelingen voor verder onderzoek. In verschillende bijlagen zijn korte www.sharehouselab.nl 5 beschrijvingen te vinden van onderwijsmodules waarin zowel de context, leerdoelen als
lesactiviteiten in worden beschreven. beschrijvingen te vinden van onderwijsmodules waarin zowel de context, leerdoelen als
lesactiviteiten in worden beschreven. Vertrekpunt en aanpak De resultaten uit het onderzoek ‘De belangrijkste logistieke skills in een veranderende
warehousecontext’ (Grolleman, Hendrickx, Warffemius, Vos, & Versluis, 2022) zijn als
vertrekpunt opgenomen. Zie voor een verkort overzicht van deze resultaten bijlage 01. Belangrijk om te benoemen is dat er binnen het MBO een keuze is gemaakt om het
MBO 1-niveau buiten beschouwing te laten. De reden hiervoor is dat een groot deel van
de competenties die hierin worden ontwikkeld algemene en een relatief klein
onderdeel logistieke competenties kent. De overige MBO-niveau’s (MBO-2/3/4) zijn
wel meegenomen. Dit sluit ook aan bij het eerdere onderzoek in het kader van de LSF-
scan (Grolleman, Hendrickx, Warffemius, Vos, & Versluis, 2022). Deze resultaten zijn besproken met Curricula Commissies (CuCo’s) &
opleidingsmanagers in medio oktober 2022. Betrokken CuCo’s op HBO niveau waren
van de Logistiek Management (LM) en Logistics Engineering (LEN) opleiding van Fontys
Techniek en Logistiek in Venlo, de CuCo van Logistiek Management (LM) en Logistics
Engineering (LEN) van Hogeschool Rotterdam en de CuCo van Logistiek Management
(LM) en Logistics Engineering (LEN) van Hogeschool Windesheim. De betrokken CuCo’s
op MBO niveau zijn het STC college in Rotterdam en Deltion te Zwolle. In de gesprekken met de CuCo’s stonden net zoals bij de expertmeetings de
analytische vaardigheden, expert op vakgebied en flexibiliteit centraal. Hierbij is ook de
rol van het bedrijfsleven besproken. Doel is om mogelijke “gaps” te signaleren in de
huidige curricula van de genoemde opleidingen en daarbij ook handreikingen te geven
hoe deze in te vullen. De gespreksverslagen met de CuCo’s, verder te noemen als
onderwijs in dit verslag, zijn indien gewenst op te vragen bij de auteurs van dit rapport. In de volgende uiteenzetting hebben we hard skills en soft skills als indeling gekozen. Hierbij zal allereerst inzicht worden gegeven in de resultaten van de interviews met de
CuCo’s vanuit het MBO en HBO, waarna een aantal (onderwijs)voorbeelden worden
benoemd die (deels) inspelen op de behoeften die vanuit de CuCo’s worden benoemd
(zie ook bijlage 2). Om vervolgens tot conclusies te komen, is het van belang om ook
het landelijke (opleidings-)profiel te evalueren vanuit de gedachte dat als het
landelijke profiel geen richting geeft, het een initiatief blijft dat vooral ingegeven wordt
door de motivatie van een individuele docent of opleiding in plaats van als sector
brede visie. Uiteindelijk zal worden afgesloten met een conclusie inclusief kritische
reflectie als ook aanbevelingen voor vervolgonderzoek. www.sharehouselab.nl www.sharehouselab.nl 6 Hard skills Hard skills
Omschrijving
HBO
R’dam
HBO
Venlo
HBO
Zwolle
MBO
R’dam
MBO
Zwolle
Data science module (bijlage 2 en 4)
X
X
nvt
nvt
Warehouse van de toekomst in relatie tot
trends in living lab (bijlage 3)
X
Management Dashboards (bijlage 2 en 4) in
power BI en Tableau
X
X
Onderzoek toepassing EXO skelets in
warehouse omgeving in living lab (pagina 5)
X
X
Sharehouse lab
-
Pick-to-light car
-
een scan systeem voor orderpicking
-
Exact online WMS pakket
-
Vision Picking door gebruik van een
head mounted display
X
X
X
X
Living Lab
-
een Pick-to-light stelling
-
een scan systeem voor orderpicking
-
een eigen ingericht ERP & WMS
systeem
-
een dozen inpak machine
X
X
X
X
Tabel x : Mogelijke gaps hard skills tussen opleidingen
Soft skills
Omschrijving
HBO
R’dam
HBO
Venlo
HBO
Zwolle
MBO
R’dam
MBO
Zwolle
Training in soft skills lab – communicatieve
vaardigheden – (bijlage 5)
X
E-logistics – Casestudy procesverbetering
(bijlage 6) en lean orange belt (bijlage 7)
X
X
Sociale innovatie binnen een lerende
organisatie
(bijlage 8)
X
Omgaan met weerstanden en draagvlak
creëren (bijlage 9)
X
Tabel x : Mogelijke gaps soft skills tussen opleidingen www.sharehouselab.nl www.sharehouselab.nl HARD SKILLS Dit gebeurt aan de hand van een
ketensimulator waarin de studenten bewust worden gemaakt van processen en hoe de
impact van de verschillende handelingen de kwaliteit of betrouwbaarheid bij een
andere partner in de keten beïnvloeden. Het denken in processen helpt om snel
verbanden te kunnen doorzien, waardoor analytische vaardigheden worden
ontwikkeld. Op het MBO wordt nog te weinig aandacht besteed aan het analyseren van grote
hoeveelheden data (o.b.v. de interviews met STC en Deltion). MBO studenten blijken
nog onvoldoende in staat om te denken in verschillende metingen die nodig zijn om
hun bevindingen en/of verbeteringen verder te kunnen onderbouwen. Deltion
benoemde daarin ook concreet dat studenten o.b.v. één meting hun bevindingen
hebben gebouwd en/of onderbouwd (n=1). Hierbij dienen de studenten (zeker op MBO
3 en MBO 4 niveau) meer kennis te hebben van de verschillende metingen die gedaan
kunnen worden, wat de voor- en nadelen zijn en hoe dit met data-analyse
onderbouwd kan worden. Het analyseren van data, door gebruik te maken van
spreadsheet-programma’s (bijv. Excel) wordt op dit moment nog niet gedaan binnen
het MBO-onderwijs (op geen enkel niveau). Vanuit analytisch perspectief wordt ‘beschrijvende statistiek’ ook niet behandeld. De
student weet wel wat het gemiddelde is, maar afwijkingen van het gemiddelde, modus
en mediaan zijn niet bekend voor de student. Vanuit Deltion wordt de Lean Orange Belt (in combinatie met Hogeschool Windesheim
te Zwolle) aangeboden. Hierin krijgen de studenten de basis mee van het werken met
Lean en een focus voor het met name het aanpakken van (kleinere) problemen
conform de DMAIC cyclus (Define, Measure, Analyse, Improve & Control) waarbij in
iedere fase een aantal tools en/of technieken worden benoemd waarmee de student
zijn probleem kan aanpakken, structuren en/of ontwerpen. Dit gebeurt op op MBO 4
niveau bij de opleiding Manager Transport en Logistiek. Na het doornemen van de gehanteerde definitie van analytische vaardigheden wordt
door het HBO onderwijs benadrukt dat ook het ‘onderzoekend vermogen’ expliciet aan
de definitie toegevoegd moet worden. Vanuit het bedrijfsleven is de behoefte aan een
sterkere inbedding van analytische vaardigheden gezien de steeds grotere
hoeveelheden data en de bijbehorende complexiteit. Met andere woorden, een nog
sterkere nadruk op data gedreven analyse. Hierbij is het niet zo dat de student een
expert moet worden op het gebied van ICT en automatisering. De student heeft wel
basiskennis van ICT nodig. HARD SKILLS Zoals eerder benoemd wordt in dit rapport een onderscheid gemaakt tussen hard- en
soft skills. In dit deel zullen we de hard skills bespreken. Tijdens de gesprekken met de CuCo’s van de verschillende onderwijsinstellingen is een
nadruk gelegd op ‘analytische vaardigheden’ en ‘expert op eigen vakgebied’. Deze
focus is aangebracht naar aanleiding van de resultaten vanuit eerder verricht
onderzoek naar de veranderende skills in een innovatieve warehouse context
(Grolleman, Hendrickx, Warffemius, Vos, & Versluis, 2022). Tijdens alle gesprekken met
de CuCo’s is gebleken dat zowel in de gesprekken met de MBO als de HBO opleidingen
een sterke focus heeft gelegen op de ‘analytische vaardigheden’. In het vervolg wordt voor zowel de ‘analytische vaardigheden’ als voor ‘expert op eigen
vakgebied’ gestart met een samenvatting van de inzichten op zowel MBO als HBO
niveau aan de hand van de gesprekken met de CuCo’s. Vervolgens wordt inzicht
gegeven in voorbeelden hoe de verschillende onderwijsinstellingen hieraan een
invulling geven. Analytische vaardigheden In het onderzoek naar de veranderende skills in een innovatieve warehouse context
(Grolleman, Hendrickx, Warffemius, Vos, & Versluis, 2022) zijn ‘analytische
vaardigheden’ gedefinieerd als: “Interpreteert complexe informatie, doorziet
verbanden, lost gestructureerd problemen op. Kan processen analyseren en storingen
signaleren en/of verhelpen.” Hieronder zal ingegaan worden op de verschillende aspecten van zowel het MBO, het
HBO en verschillende voorbeelden vanuit het onderwijs. Resultaten MBO CuCo s
Vanuit de 3 MBO niveaus (2/3/4) is het goed om op voorhand het onderscheid en de
focus op analytische vaardigheden verder te duiden. Hierbij wordt door verschillende
geïnterviewden vanuit verschillende CuCu’s het volgende benoemd;
Niveau
Toelichting
MBO 1
Voert uit
MBO 2
Signaleert
MBO 3
Pak problemen op / herkent complexere
problemen ¹
MBO 4
Verbanden zien en oplossen / pakt
complexere problemen op
1 Dit zijn problemen die verder gaan dan het ‘eigen domein’. Zo wordt het effect van problemen en acties doorzien
bijvoorbeeld in de IT, Materieel, toeleveranciers, KPI omschrijvingen, etc. 1 Dit zijn problemen die verder gaan dan het ‘eigen domein’. Zo wordt het effect van problemen en acties doorzien
bijvoorbeeld in de IT, Materieel, toeleveranciers, KPI omschrijvingen, etc. www.sharehouselab.nl 8 Vanuit het STC wordt aanvullend benoemd dat er veel wordt ingezet op het
‘procesdenken’. Het zien van verbanden tussen verschillende lagen in een organisatie
alsook voor een simpele logistieke keten. www.sharehouselab.nl HARD SKILLS Het is belangrijk dat de student de vaardigheid beheerst om
de logistieke parameters uit datasets te halen. Hiervoor moet de student eerst op zoek
gaan naar deze parameters. De uitdaging is om samen met het bedrijfsleven een
complexe en uitdagende dataset te verstrekken die studenten kunnen analyseren om www.sharehouselab.nl 9 van daaruit te kunnen simuleren. De huidige datasets in het onderwijs zijn nog te vaak
te theoretisch. van daaruit te kunnen simuleren. De huidige datasets in het onderwijs zijn nog te vaak
te theoretisch. Gaps & invulling Het interpreteren van complexe informatie is een HBO skill die ook raakt aan MBO 4. Voor alle MBO niveaus is kritisch vermogen belangrijk en het durven en kunnen melden
van mogelijke verbeteringen (kun je de boodschap overbrengen). Alle MBO niveaus en
het HBO niveau hebben samen een rol in het signaleren en oplossen van een probleem. Een belangrijke analytische vaardigheid voor MBO 2 is het signaleren en kunnen
melden. Bijvoorbeeld het signaleren als je ziet dat jouw order te laat wordt uitgeleverd
en de informatie hierover juist en compleet melden aan de juiste persoon in de
organisatie. Een belangrijke analytische vaardigheid voor MBO 3 is het herkennen van
complexere problemen en deze goed en tijdig overdragen aan de juiste persoon in de
organisatie die de problemen kan structureren en analyseren. Een belangrijke
analytische vaardigheid voor MBO4 is verbanden zien en oplossingen vinden. Op MBO
4 niveau is in het curriculum veel aandacht voor het juist en compleet kunnen melden
(communicatie vaardigheden), de stakeholders, jouw rol in de keten, vakoverstijgend
kunnen denken en werken en procesanalyse. Het geheel benoemen en zien in het
‘procesdenken’ lijkt daarin meer terug te komen vanuit verschillende (deel)
onderwerpen maar niet samengebracht tot één geheel. Daarnaast zouden er in een living lab omgevingen concrete experimenten kunnen
worden uitgevoerd. Bijvoorbeeld eerst HBO studenten laten nadenken hoe ze de meest
efficiënte route voor orderpicken kunnen ontwerpen. Een keer op basis van eigen
inzicht en een keer op basis van literatuur. Daarna kunnen beide methoden in het
Sharehouse worden toegepast en kunnen pick tijden, pick fouten, loop afstanden, etc. worden gemeten en kunnen beide systemen worden vergeleken. Zowel MBO als HBO
studenten kunnen hierbij een rol vervullen. Een ander voorbeeld kan zijn dat de HBO
studenten een werkinstructie ontwerpen, deze in het Sharehouse laten uitvoeren door
MBO studenten en vervolgens verbeteren op basis van feedback. Deze feedback kan
bijvoorbeeld komen van MBO studenten die volgens de werkinstructie hebben gewerkt
of uitgevoerde metingen van het aantal pick fouten of tijden aan de hand ven
registraties. Vervolgens kan de werkinstructie aangepast worden om tenslotte op basis
van metingen en feedback te beoordelen of het na de aanpassingen werkelijk beter
gaat. Conclusie www.sharehouselab.nl
Conclusie
Kort samengevat kan worden geconcludeerd dat er een volgende behoefte is vanuit
MBO en HBO opleidingen op de skill ‘analytische vaardigheden’. Behoefte onderwijs
‘Analytische vaardigheden’
MBO
HBO
Meer procesdenken
X
Data-analyses in Excel
X
X
De verbetercyclus
X
Logistieke parameters
X ort samengevat kan worden geconcludeerd dat er een volgende behoefte is vanui
BO en HBO opleidingen op de skill ‘analytische vaardigheden’. Kort samengevat kan worden geconcludeerd dat er een volgende behoefte is vanuit
MBO en HBO opleidingen op de skill ‘analytische vaardigheden’. www.sharehouselab.nl
Behoefte onderwijs
‘Analytische vaardigheden’
MBO
HBO
Meer procesdenken
X
Data-analyses in Excel
X
X
De verbetercyclus
X
Logistieke parameters
X X www.sharehouselab.nl www.sharehouselab.nl 10 Concrete praktijkvraagstukken
zelf ervaren en ontwerpen Concrete praktijkvraagstukken
zelf ervaren en ontwerpen www.sharehouselab.nl Expert op vakgebied g
In het onderzoek naar de veranderende skills in een innovatieve warehouse context
(Grolleman, Hendrickx, Warffemius, Vos, & Versluis, 2022) is ‘expert op eigen
vakgebied’ gedefinieerd als: “Diepgaande kennis, continu op zoek naar nieuwe kennis
op eigen vakgebied. Meer specifiek de kennis van diverse tools in een warehouse en/of
deze kunnen toepassen.” Vanuit deze skill is in de gesprekken een nadruk komen te
liggen op enerzijds de gebruikte tools en anderzijds de toepassing van deze tools voor
studenten binnen het onderwijs. Hieronder zal ingegaan worden op de verschillende aspecten van zowel het MBO, het
HBO en verschillende voorbeelden vanuit het onderwijs. 11 Als onderdeel van ‘expert op eigen vakgebied’ is ook gesproken over het ‘nieuwsgierig’
krijgen van de MBO-studenten. Hoe zorgen de student er nu voor dat zij zich ook na
hun studie, vanuit hun eigen regie, blijven ontwikkelen. Geïnitieerd door Deltion en
onderschreven door het STC geven beiden aan dat dit in het reguliere onderwijs nog
weinig gebeurt. Er zitten weinig onderdelen in waarin de studenten zelf inhoud moeten
zoeken. Veel, zo niet alle inhoud wordt voorgeschreven voor de student. Hierin zou
meer vrijheid en eigen initiatief bevorderd moeten worden in het onderwijs. De term “expert” doet binnen het MBO meer denken aan theorie. Echter MBO niveau 1
en 2 zijn vooral op de praktijk gericht. Op MBO 1 en 2 niveau gaat het om praktische
vaardigheden op de werkvloer. MBO niveau 1 krijgt gestuurde instructies en voert deze
uit. MBO 2/3 niveau zou de ideeënbus kunnen vullen met gesignaleerde
verbetermogelijkheden en MBO 4 zou deze uit kunnen voeren. In het curriculum gaat
het om de juiste balans tussen theorie en praktijk. Deze ligt per niveau (1, 2, 3 of 4)
anders. Op dit moment zijn praktijkopdrachten in het Sharehouse bij het STC nog vaak
ingeschaald op niveau 3 en 4 waarbij gewerkt wordt a.d.h.v. lijsten met vragen en
instructies. Op niveau 1 en 2 zou nog meer met praktijkopdrachten in het Sharehouse
kunnen worden gewerkt zonder de lijsten met vragen en instructies maar juist door de
goede vragen te stellen terwijl de studenten praktisch aan het werk zijn. Heel belangrijk
daarbij zijn goede praktijkbegeleiders die de juiste vragen kunnen stellen. Om ‘expert’ te zijn en te blijven wordt vanuit Deltion en STC aangegeven om meer
onderwijs te willen ontwikkelen waarbij MBO en HBO – studenten gezamenlijk aan
producten werken. Hier doel hierin is vooral te zien en ervaren wat de verschillen zijn
tussen de opleidingsniveaus en daarop te acteren in zowel kennisniveau als ook deels
in houding, communicatie en gedrag. Daarnaast wordt ook concreet de rol van het bedrijfsleven benoemt. Het bedrijfsleven
zou praktijkopdrachten en goede praktijkbegeleiders kunnen leveren waar MBO
studenten mee aan de slag kunnen in de veilige en gecontroleerde Living Lab
omgeving. Het liefst brede opdrachten waar de diverse aspecten van de logistiek in
samenhang aan bod komen, onder andere over digitalisering. Een deel van de theorie
kan dan nog beter in het Sharehouse worden gegeven waardoor de koppeling tussen
theorie en praktijk nog duidelijker wordt. Resultaten MBO-CuCo’s Binnen het Deltion wordt gebruikt gemaakt van Exact Online waarbij op verschillende
opleidingsniveaus diverse handelingen worden toegepast. Zo dient de MBO 3
medewerker pakbonnen te kunnen verwerken in een WMS/ERP systeem en dient een
MBO 4 medewerker de pakbonnen ook te kunnen samenstellen en uitprinten en
verschillende (simpele)foutieve registraties te kunnen corrigeren. Hierbij komt wel de
vraag in hoeverre het MBO kan bijblijven bij de innovaties van vandaag de dag. Living
Lab – Initiatieven als Sharehouse (STC Rotterdam) en Logistics Lab (Deltion Zwolle)
worden daarbij wel als concrete voorbeelden benoemd om hier handen en voeten aan
te geven. Binnen Deltion bestaat de innovatie momenteel uit; -
een Pick-to-light stelling
-
een scan systeem voor orderpicking
-
een eigen ingericht ERP & WMS systeem
-
een dozen inpak machine
-
er wordt nagedacht over facilitering van een Exoskelet Binnen STC bestaat de innovatie momenteel uit; Binnen STC bestaat de innovatie momenteel uit; •
Pick-to-light car •
een scan systeem voor orderpicking •
Exact online WMS pakket •
Er wordt nagedacht over facilitering van een Exoskelet •
Vision Picking door gebruik van een head mounted display Zoals eerder benoemd, wordt binnen het MBO geen gebruik gemaakt van specifieke
data analyse tools die hierin kunnen ondersteunen. www.sharehouselab.nl 12 Als laatste wordt de rol van begeleiding en het niveau van praktijkopdrachten
benoemt. Het bedrijfsleven heeft, zeker in het MBO, samen met het onderwijs een
essentiële rol in het opleiden van de studenten. Het is belangrijk dat een MBO student
een kwalitatief goede stageopdracht en begeleiding heeft waarbij de verschillende
facetten van de logistiek en de breedte van de werkzaamheden in samenhang aan bod
komen. Hierin wordt ook benoemt dat het niveau van zowel de begeleiding als de
opdrachten nog met regelmatig discutabel zijn. Zo werken MBO studenten, zeker op
niveau 3 en 4, nog regelmatig mee in het operationele proces terwijl zij juist meer
besturende en regelende taken zouden moeten (kunnen) uitvoeren. Binnen het HBO wordt gewerkt met verschillende tools, zowel vanuit data-analyse als
vanuit bedrijfsadministratie (ERP). De data analyse tools worden op HBO niveau nog vaak vanuit de theorie ingezet terwijl
juist toepassing in een echte praktijkomgeving heel belangrijk is. Nu blijft het vaak bij
een bedrijfsbezoek of gastcollege. Daarnaast moet een student meer zelf op zoek leren
gaan naar expertise. Daarvoor moet ook bij de HBO student voortdurend een actieve
onderzoekende houding gestimuleerd worden. De HBO Student moet nog meer
getraind worden om zelf op zoek te gaan naar de juiste expertise om zich deze kennis
zichzelf vervolgens ook eigen te maken. Tools die steeds meer gebruikt worden in het bedrijfsleven en ook in het onderwijs
voor visualisatie van data zijn PowerBi (Microsoft) en Tableau. Tools die gebruikt
worden voor structurering en oorzaken analyse zijn bijvoorbeeld het Green Belt
trainingsprogramma. Waar het vaak nog aan ontbreekt zijn een WMS of een TMS
omgeving met een dataset vanuit het bedrijfsleven en met een koppeling tussen een
WMS en TMS. 11 Hierbij is het belangrijk dat
praktijkbegeleiders uit het werkveld ervaring hebben met ook MBO 1 en 2 niveau. De
praktijkbegeleider moet op het juiste moment de juiste vragen weten te stellen terwijl
de student praktisch aan het werk is. Vanuit dit perspectief denkende wordt aangegeven dat het bedrijfsleven nog
onvoldoende bekend is met de rol die Living Labs kunnen betekenen voor het
werkveld. De meerwaarde van Living Labs voor het werkveld kan nog beter over het
voetlicht worden gebracht. Soms is het bedrijfsleven huiverig ontwikkelingen te delen
om concurrentiele voordelen te behouden. Deze hindernis zou kleiner kunnen zijn
tussen bedrijven uit verschillende bedrijfstakken die niet elkaars concurrent zijn. Deze
bedrijven zouden in een Living Lab samen met studenten en docenten kunnen werken
(testen, ontwikkelen, leren, onderzoeken). www.sharehouselab.nl Voorbeelden van ‘eigen’ initiatieven Voorbeelden van ‘eigen’ initiatieven
Alle CuCo’s gaven aan op verschillende elementen al druk bezig te zijn met het
ontwikkelen van onderwijsmateriaal dat meer en beter aansluit bij eerder
gedefinieerde behoeften. Hieronder zullen een aantal van deze initiatieven worden
gedeeld waarbij deze in de tabel hieronder worden gekoppeld aan de behoeften zoals
hiervoor is opgesteld. Behoefte onderwijs
MBO HBO
Een
toekomstbestendig
warehouse
Deltion E-Logistics
Experimenteren in
Living Lab: exoskelets
Datadriven Logistics
Analytische vaardigheden
Meer procesdenken
X
X
Data-analyses in Excel
X
X
X
De verbetercyclus
X
X
Logistieke parameters
X
X
X
Concrete
praktijkvraagstukken
zelf ervaren en
ontwerpen
X
X
X
Expert op eigen vakgebied
Werken met ERP
systemen
X
X
Werken met BI-tools
X
X
Bevorderen eigen
initiatief
X
X
X
X
Onderwijs MBO en
HBO combineren
X
X
X
X
Concrete
praktijkvraagstukken
X
X
X
X
X
Green Belt
trainingsprogramma’s
X
Een toekomstbestendig warehouse
Zo speelt bij het STC in Rotterdam een traject waarbij studenten vanuit de Hogeschool
Rotterdam aan een project werken genaamd ‘Ontwerpen van een toekomstbestendig
warehouse (zie bijlage 4). Hierin zijn de Leng -studenten van de HR gedurende een
aantal dagdelen aanwezig binnen het Sharehouse van het STC om vooral de innovaties
te zien en ervaren. Dit kunnen zij naar een inrichting en ontwerp van een nieuw Alle CuCo’s gaven aan op verschillende elementen al druk bezig te zijn met het
ontwikkelen van onderwijsmateriaal dat meer en beter aansluit bij eerder
gedefinieerde behoeften. Hieronder zullen een aantal van deze initiatieven worden
gedeeld waarbij deze in de tabel hieronder worden gekoppeld aan de behoeften zoals
hiervoor is opgesteld. Behoefte onderwijs
MBO HBO
Een
toekomstbestendig
warehouse
Deltion E-Logistics
Experimenteren in
Living Lab: exoskelets
Datadriven Logistics Conclusie www.sharehouselab.nl
Kort samengevat kan worden geconcludeerd dat er een volgende behoefte is vanuit
MBO en HBO opleidingen op de skill ‘expert op eigen vakgebied’. Behoefte onderwijs
‘Expert op eigen vakgebied’
MBO
HBO
Werken met ERP systemen
X
X
Werken met BI-tools
X
Bevorderen eigen initiatief
X
Onderwijs MBO en HBO combineren
X
X
Concrete praktijkvraagstukken
X
X
Green Belt trainingsprogramma’s
X Kort samengevat kan worden geconcludeerd dat er een volgende behoefte is vanuit
MBO en HBO opleidingen op de skill ‘expert op eigen vakgebied’. Kort samengevat kan worden geconcludeerd dat er een volgende behoefte is vanuit
MBO en HBO opleidingen op de skill ‘expert op eigen vakgebied’. Kort samengevat kan worden geconcludeerd dat er een volgende behoefte is vanuit
MBO en HBO opleidingen op de skill ‘expert op eigen vakgebied’. Behoefte onderwijs
‘Expert op eigen vakgebied’
MBO
HBO
Werken met ERP systemen
X
X
Werken met BI-tools
X
Bevorderen eigen initiatief
X
Onderwijs MBO en HBO combineren
X
X
Concrete praktijkvraagstukken
X
X
Green Belt trainingsprogramma’s
X www.sharehouselab.nl 13 www.sharehouselab.nl Een toekomstbestendig warehouse g
Zo speelt bij het STC in Rotterdam een traject waarbij studenten vanuit de Hogeschool
Rotterdam aan een project werken genaamd ‘Ontwerpen van een toekomstbestendig
warehouse (zie bijlage 4). Hierin zijn de Leng -studenten van de HR gedurende een
aantal dagdelen aanwezig binnen het Sharehouse van het STC om vooral de innovaties
te zien en ervaren. Dit kunnen zij naar een inrichting en ontwerp van een nieuw www.sharehouselab.nl 14 warehouse. In de toekomst zijn plannen om dit uit te breiden waarbij MBO studenten
ook een rol gaan spelen bij het experimenteren en testen van mogelijke ontwerpen. warehouse. In de toekomst zijn plannen om dit uit te breiden waarbij MBO studenten
ook een rol gaan spelen bij het experimenteren en testen van mogelijke ontwerpen. De achterliggende gedachte is dat de logistieke sector te maken heeft met grote
ontwikkelingen en innovaties. Denk aan alle ontwikkelingen op het gebied van ICT zoals:
Internet of Things (IoT), sensor technologie, robots, virtual reality (VR), augmented
reality (AR), of big data. HBO-studenten zullen steeds vaker komen te werken in een
digitale omgeving. Het gaat erom in dit project, hoe het warehouse toekomstbestendig
te maken is zodat het mee kan in de logistieke ontwikkelingen van de toekomst. En
technologie helpt daarbij. In een toekomstbestendig warehouse zijn de goederen- en
informatiestroom met elkaar verbonden. Het warehouse van de 21e eeuw is een high-
tech hal waarin veel geautomatiseerd is. De student gaat in dit project op zoek naar
logistieke trends en zet deze om in een warehouse ontwerp. Hierbij vinden er een
aantal lessen plaats in het living lab van Sharehouse bij STC. Dit stimuleert studenten
om out-of-the-box te denken en ook gelijk hardware en software in de praktijk te zien
en te ervaren. Experimenteren in een Living Lab: Exoskelets Hierbij wordt geëxperimenteerd met verschillende vormen van innovatieopdrachten in
de praktijklocatie van het MBO. Zo heeft er een onderzoek plaats gevonden vanuit het
Zwolse initiatief Logistics Lab waarbij studenten de mogelijkheden van Exoskelets
hebben uitgezocht voor een logistieke operatie. Hierbij hebben de MBO studenten ook
weer gebruik kunnen maken van de aanwezige Exoskelets waarbij HBO studenten een
deel van de uitleg en toelichting op zich hebben genomen. Deze initiatieven worden door alle betrokken partijen ondersteundmet de oproep om
meer niveau-overstijgend onderwijs te ontwikkelen.,waarbij zowel niveau MBO2/3/4 als
HBO betrokken is. Bovenstaande voorbeelden zijn hier een toepssing van, maar zou ook
binnen het MBO (met de verschillende opleidingsniveaus) meer opgepakt moeten
worden ten einde de complexiteit van verschillende problemen gezamenlijk te kunnen
aanpakken. www.sharehouselab.nl Deltion E-Logistics Vanuit het Deltion speelt op het vlak van het uitvoeren van een business case. Hierbij is
rondom de leeractiviteiten van MBO 2 en MBO 3 studenten een casus (Deltion E-
logistics) ontwikkeld. Hierbij zijn MBO 2 en MBO 3 studenten de medewerkers van het
bedrijf die actief orders verzamelen. De HBO studenten hebben de opdracht om
gezamenlijk met de MBO studenten te komen tot procesverbeteringen die de
‘grootste’ impact hebben op het reduceren van de doorlooptijd, rekening houdend met
de eenvoud van het proces aangezien de medewerkers het gehele proces simpel
moeten kunnen blijven begrijpen. Hierin vindt dus overleg plaats tussen de zowel de
studenten van het MBO als het HBO. Datadriven Logistics Na het doornemen van de curricula van de HBO opleiding Logistics engineering van
Hogeschool Rotterdam en Fontys Techniek en Logistiek in Venlo zijn nieuwe www.sharehouselab.nl 15 toepassingen naar voren gekomen die inspelen op de Datagedreven vaardigheden van
een student (zie bijlagen 3 en 5). Het is op zich logisch dat deze vaardigheden en
toepassingen sterker terug te vinden zijn bij de Logistics Engineering opleidingen dan
bij de Logistiek Management opleiding. Deze laatste opleiding is de nadruk sterker op
het organiseren en managen van logistieke processen. Alhoewel ook voor deze
opleiding de gevraagde analytische vaardigheden op bijv. data sets belangrijker zijn
geworden. Hier komen we in de kritische reflectie en aanbevelingen op terug. toepassingen naar voren gekomen die inspelen op de Datagedreven vaardigheden van
een student (zie bijlagen 3 en 5). Het is op zich logisch dat deze vaardigheden en
toepassingen sterker terug te vinden zijn bij de Logistics Engineering opleidingen dan
bij de Logistiek Management opleiding. Deze laatste opleiding is de nadruk sterker op
het organiseren en managen van logistieke processen. Alhoewel ook voor deze
opleiding de gevraagde analytische vaardigheden op bijv. data sets belangrijker zijn
geworden. Hier komen we in de kritische reflectie en aanbevelingen op terug. Een voorbeeld dat hierop inspeelt is van de Logistics Engineering opleiding van Fontys
Hogeschool Techniek en Logistiek in Venlo. Hier ontwerpen studenten na het
formuleren van KPI’s voor verschillende afdelingen in een logistieke onderneming een
dashboard met behulp van PowerBI. Hiervoor moeten ze de aangereikte dataset
opschonen en relaties aanbrengen om vervolgens een dashboard te bouwen. Hiervoor
worden ze ook getraind op het maken van krachtige visualisaties. Op deze wijze
kunnen ze een krachtig instrument ontwikkelen voor een bijvoorbeeld steeds
complexere warehouse omgeving. Tenslotte hebben de HR en Fontys opleidingen een
datascience module opgezet waarbij data analyse tools als R ingezet worden. Een
uitgebreidere toelichting van genoemde toepassingen zijn opgenomen in bijlagen 03
en 05. SOFT SKILLS Vanuit de resultaten van het eerder gepubliceerde onderzoek (Grolleman, Hendrickx,
Warffemius, Vos, & Versluis, 2022) is met name geconstateerd dat het bedrijfsleven in
een technologie veranderende warehouseomgeving behoefte heeft aan studenten die
met name ‘respectvol’ met elkaar omgaan, elkaar durven aan te spreken op gedrag en
de kwaliteit en kwantiteit van het geleverde werk. Tevens is benoemd dat studenten
bewust moeten zijn van de eigen kennis en vaardigheden en daarin ook hun eigen
ontwikkeling. In het vervolg worden soft skills in breed perspectief samengevat aan de hand van de
CuCu gesprekken op zowel MBO als HBO niveau. Vervolgens wordt inzicht gegeven in
voorbeelden hoe de verschillende onderwijsinstellingen hieraan een invulling geven en
inzicht gegeven in de behoeften vanuit het onderwijs. Resultaten CuCo’s HBO Ook op HBO niveau is het zoeken naar de vorm waarin ‘flexibiliteit’ wordt aangeboden. Binnen Windesheim wordt hierbij gebruik gemaakt van een leerlijn rondom
Professionele ontwikkeling. In het eerste jaar wordt er met de studenten gekeken naar
de rol die zij zelf vervullen in verschillende samenwerkingsvormen, waarop zij moeten
reflecteren. Daarin staat het onderwerp aanpassingsvermogen ook centraal. Tegelijkertijd zien we ook hierin dat het echt omgaan met snelle veranderingen en
daarin vooral denken in mogelijkheden en minder in problemen nog een punt is
waaraan ook HBO-opleidingen kunnen werken. Eén van de andere geconstateerde uitdagingen binnen het HBO-onderwijs is het
omarmen van sociale innovatie en hoe hierin HBO-studenten geschoold kunnen
worden. Veranderingen gaan immers sneller en sneller, de technologie van vandaag is
morgen alweer achterhaald. Hoe ga je hier dan mee om als individu én organisatie. Immers het weten wat de technologie en innovaties zijn van vandaag, is morgen alweer
ouderwets. Kortom, het HBO dient hierin studenten te triggeren en te leren hoe
studenten hierin zelfstandig en intrinsiek gemotiveerd kunnen raken om te leren alsook
hoe een cultuur binnen een organisatie kan worden gevormd om hier op een goede
manier mee om te gaan. Resultaten CuCo’s MBO Vanuit de MBO opleidingen wordt benadrukt dat het werken aan soft skills relatief
weinig gebeurt. Natuurlijk wordt er binnen de verschillende opleiding samengewerkt,
maar relatief weinig gereflecteerd op hun eigen houding en gedrag aspecten. Voor veel
MBO studenten is deze stap ook lastiger, aangezien zij deze punten graag in de praktijk
willen oppakken. Hierin is dan ook de vraag vanuit de MBO scholen op welke manier de
soft skills meer opgepakt kunnen worden aan de hand van concrete casusopdrachten
vanuit bedrijven in de regio. Daarnaast wordt benoemd dat het een goede stap zou zijn als de studenten meer en
bewuster communiceren met studenten vanuit verschillende opleidingsniveaus. Hierbij www.sharehouselab.nl 16 wordt niet alleen gedoeld op MBO en HBO, maar ook op MBO onderling. Hoe vaak zijn
MBO4 en MBO1 studenten concreet en gezamenlijk bezig met een opdracht waarin de
sterktes van beide groepen studenten onderzocht moeten worden om tot een goed
eindproduct te komen? Ook hier wordt het ‘procesdenken’ weer benoemt. Niet vanuit
de kennis die je dient te hebben over het proces, maar juist vanuit het aansluiten bij
het kennisniveau van je collega en wat ga je doen als dit kennisniveau niet aansluit. Ben
je dan in staat om anders te gaan communiceren? Als laatste onderdeel waarover veel is gesproken vanuit de CuCo’s op MBO niveau is
‘flexibiliteit’. Hierbij draait het met name om het snel kunnen aanpassen bij tegenslag of
weerstand. Flexibiliteit vraagt veel van de communicatieve vaardigheden van de
studenten. Binnen het Deltion wordt hierin nog weinig aandacht aan gegeven en is met
name afhankelijk van de docent. Sommige docenten geven wel heel bewust kritische
feedback, om de studenten te laten ervaren dat de praktijk niet altijd positief en
opbouwend is. Reflectie hierop, samen met de student vindt nauwelijks plaats. Het
geheel is daarbij nu sterk afhankelijk van eigen initiatief en inzet van de docent. Verder geldt dat alle geïnterviewde CuCo’s benoemen dat soft skills in het curriculum
naar voren komt, in de vorm van reflecties tijdens praktijkopdrachten en stages. Daarmee blijft het wel bedrijfs-, context- en situatie afhankelijk in hoeverre alle
studenten hier op een vergelijkbaar niveau mee te maken hebben gekregen. www.sharehouselab.nl Voorbeelden van ‘eigen’ initiatieven Alle CuCo’s gaven aan op verschillende elementen al druk bezig te zijn met het
ontwikkelen van onderwijsmateriaal dat meer en beter aansluit bij eerder
gedefinieerde behoeften. Hieronder zullen een aantal van deze initiatieven worden
gedeeld waarbij deze in de tabel hieronder worden gekoppeld aan de behoeften zoals
hiervoor is opgesteld. Behoefte onderwijs
MBO HBO
Het soft skills lab
Deltion E-Logistics
Leerlijn Professionele
ontwikkeling
De lerende organisatie SOFT SKILLS
Concrete
praktijkvraagstukken
X
X
Communiceren met
andere
onderwijsniveaus
X
X
Flexibele houding
X
X
X
Nieuwsgierigheid
X
X
X Conclusie Kort samengevat kan worden geconcludeerd dat er een volgende behoefte is vanuit
MBO en HBO opleidingen op het gebied van soft skills in meer algemene zin. www.sharehouselab.nl 17
Behoefte onderwijs
Soft Skills in algemene zin
MBO
HBO
Concrete praktijkvraagstukken
X
Communiceren met andere
onderwijsniveaus
X
Flexibele houding
X
x
Nieuwsgierigheid
X Voorbeelden van ‘eigen’ initiatieven
Alle CuCo’s gaven aan op verschillende elementen al druk bezig te zijn met het
ontwikkelen van onderwijsmateriaal dat meer en beter aansluit bij eerder
gedefinieerde behoeften. Hieronder zullen een aantal van deze initiatieven worden
gedeeld waarbij deze in de tabel hieronder worden gekoppeld aan de behoeften zoals
hiervoor is opgesteld. Behoefte onderwijs
MBO HBO
Het soft skills lab
Deltion E-Logistics
Leerlijn Professionele
ontwikkeling
De lerende organisatie
SOFT SKILLS
Concrete
praktijkvraagstukken
X
X
Communiceren met
andere
onderwijsniveaus
X
X
Flexibele houding
X
X
X
Nieuwsgierigheid
X
X
X
Het Soft Skills Lab
Vanuit het STC is het Soft Skills Lab ontwikkeld. Hierin worden studenten aan de hand
van praktijksituaties uitgedaagd om de juiste (communicatieve) vaardigheden in te
zetten ten einde het gewenste resultaat te behalen. De studenten worden getraind in
het omgaan met situaties en de (sociale) vaardigheden die dit van hen vraagt. Hierbij
wordt samengewerkt met verschillende bedrijven uit de regio die hun specifieke
praktijksituaties benoemen. In sommige gevallen zijn studenten van de toneelopleiding
betrokken bij het uitvoeren van opdrachten om de STC studenten een zo’n echte
mogelijke situatie te laten ervaren. Een voorbeeld hoe het wordt toegepast vanuit het
onderwijs wordt uiteengezet in bijlage 06. www.sharehouselab.nl Deltion E-Logistics Vanuit de samenwerking tussen Deltion (MBO) en Windesheim (HBO) is een case
ontwikkeld waarin de studenten van Deltion die de uitvoering van magazijntaken aan
het leren zijn, worden meegenomen door HBO-studenten ten einde procesverbetering
te realiseren. Natuurlijk zitten in de casus van de MBO-studenten verschillende ‘fouten’
verweven om daarmee het leerproces te voor de MBO-studenten te vergroten. Tegelijkertijd kunnen de HBO studenten met hun eigen vaardigheden, tools en
technieken de MBO studenten ondersteunen in de manier waarop zij deze Tegelijkertijd kunnen de HBO studenten met hun eigen vaardigheden, tools en
technieken de MBO studenten ondersteunen in de manier waarop zij deze
constateringen kunnen aanpakken en verder brengen. Het doel dat hiermee
nagestreefd wordt is tweezijdig; -
De MBO-studenten leren dat zij zelf kritisch mogen zijn op de uitgevoerde
processen en hiervoor (kleinere) verbeteringen kunnen en mogen aandrag De MBO studenten leren dat zij zelf kritisch mogen zijn op de uitgevoerde
processen en hiervoor (kleinere) verbeteringen kunnen en mogen aandragen
-
De HBO-studenten leren door de medewerkers vanaf het begin direct te
betrekken, verbeteringen makkelijker en eenvoudiger gedragen en daarmee ook
geadopteerd kunnen worden. -
De HBO-studenten leren door de medewerkers vanaf het begin direct te
betrekken, verbeteringen makkelijker en eenvoudiger gedragen en daarmee ook
geadopteerd kunnen worden. Voor meer informatie hierover zie bijlage 07 Voor meer informatie hierover zie bijlage 07 Vanuit de samenwerking tussen Deltion en Windesheim in de casus is ook de wens
ontstaan om de MBO-studenten meer handvatten en tools te geven om zelf meer regie
te kunnen pakken in het verbeteren van processen in kleine stappen. Hiermee worden
de MBO – studenten dus ook, mede door de activiteiten in de casus, geconfronteerd
met het verbeteren van vaardigheden. Hiervoor is een training opgezet waarin de Lean
Orange Belt binnen het MBO wordt opgepakt. Hierin leren de studenten de eerste
vaardigheden rondom het gestructureerd werken aan continu kleine verbeteringen. Aanvullende informatie hierover is te vinden in bijlage 08 Leerlijn Professionele ontwikkeling Om inzicht te krijgen in de wijze waarop studenten zelf bewust zijn van hun eigen
gedrag en houding wordt vanuit Windesheim vanaf jaar 1 gestart met een professionele
leerlijn. Hierin worden de studenten continu gereflecteerd op hun eigen houding en
gedrag in verschillende situaties om op deze manier inzicht te krijgen in de eigen
sterktes en zwaktes. De gedachte is dat samenwerking key is in menig organisatie,
kortom het is (en blijft) van groot belang je eigen sterktes en uitdagingen te kennen en
deze van anderen ook (snel) te herkennen zodat een effectieve wijze van
samenwerking aangegaan kan worden. Zie bijlage 10 voor een voorbeeld van de
invulling zoals dit vanuit Windesheim is vormgegeven. www.sharehouselab.nl Het Soft Skills Lab Vanuit het STC is het Soft Skills Lab ontwikkeld. Hierin worden studenten aan de hand
van praktijksituaties uitgedaagd om de juiste (communicatieve) vaardigheden in te
zetten ten einde het gewenste resultaat te behalen. De studenten worden getraind in
het omgaan met situaties en de (sociale) vaardigheden die dit van hen vraagt. Hierbij
wordt samengewerkt met verschillende bedrijven uit de regio die hun specifieke
praktijksituaties benoemen. In sommige gevallen zijn studenten van de toneelopleiding
betrokken bij het uitvoeren van opdrachten om de STC studenten een zo’n echte
mogelijke situatie te laten ervaren. Een voorbeeld hoe het wordt toegepast vanuit het
onderwijs wordt uiteengezet in bijlage 06. Vanuit het STC is het Soft Skills Lab ontwikkeld. Hierin worden studenten aan de hand
van praktijksituaties uitgedaagd om de juiste (communicatieve) vaardigheden in te
zetten ten einde het gewenste resultaat te behalen. De studenten worden getraind in
het omgaan met situaties en de (sociale) vaardigheden die dit van hen vraagt. Hierbij
wordt samengewerkt met verschillende bedrijven uit de regio die hun specifieke
praktijksituaties benoemen. In sommige gevallen zijn studenten van de toneelopleiding
betrokken bij het uitvoeren van opdrachten om de STC studenten een zo’n echte
mogelijke situatie te laten ervaren. Een voorbeeld hoe het wordt toegepast vanuit het
onderwijs wordt uiteengezet in bijlage 06. www.sharehouselab.nl 18 18 De lerende organisatie Vanuit de uitdaging op het gebied van nieuwsgierigheid vanuit het individu en
organisatie is binnen de opleiding Logistics Management van Windesheim is een
module ontwikkeld rondom ‘de lerende organisatie’. Dit is een organisatie die niet
alleen technologisch innovatief is, maar ook sociaal innovatief. Een organisatie
waar men flexibel is om verandering te adopteren, eigen te maken en te
realiseren. En waar op alle niveaus wordt geleerd en kennis en vaardigheden
worden ontwikkeld en gedeeld. In deze module leert de student zowel te kijken www.sharehouselab.nl 19
naar zijn eigen individu, zijn rol in de organisatie en de organisatie al geheel. Immers, alleen met een ‘lerende organisatie’ ben je sneller in staat om van
richting te veranderen en snel in te spelen op de veranderingen van vandaag én
van morgen. Deze module wordt anno 2023 als keuzemodule aangeboden, waarbij
studenten deze module met relatief veel zelfstudie zichzelf eigen maken. Nieuwsgierig
naar meer informatie zie dan bijlage 09. 19 naar zijn eigen individu, zijn rol in de organisatie en de organisatie al geheel. Immers, alleen met een ‘lerende organisatie’ ben je sneller in staat om van
richting te veranderen en snel in te spelen op de veranderingen van vandaag én
van morgen. Deze module wordt anno 2023 als keuzemodule aangeboden, waarbij
studenten deze module met relatief veel zelfstudie zichzelf eigen maken. Nieuwsgierig
naar meer informatie zie dan bijlage 09. naar zijn eigen individu, zijn rol in de organisatie en de organisatie al geheel. Immers, alleen met een ‘lerende organisatie’ ben je sneller in staat om van
richting te veranderen en snel in te spelen op de veranderingen van vandaag én
van morgen. Deze module wordt anno 2023 als keuzemodule aangeboden, waarbij
studenten deze module met relatief veel zelfstudie zichzelf eigen maken. Nieuwsgierig
naar meer informatie zie dan bijlage 09. ALGEMENE REFLECTIE ROL BEDRIJFSLEVEN Vanuit de gevoerde gesprekken met de CuCo’s van zowel de MBO als HBO opleidingen
is gebleken dat concrete praktijkvraagstukken een duidelijke wens is vanuit het
onderwijs als geheel. Hieronder volgen nog een aantal verschillende aandachtspunten
op het gebied van het bedrijfsleven. Visie MBO De rol van het bedrijfsleven is van essentieel belang voor de MBO opleidingen. In de
praktijk lijkt het dat de ontwikkeling van studenten afhangt van het bedrijf waar de
betreffende student stage loopt. Loop je stage bij een bedrijf met veel en goede
begeleiding , dan leer je meer. De vraag is op welke manier hier bij toekenning van
opleidingserkenning (dit verloopt via het SBB) meer aandacht aan gegeven kan worden
en hoe er, na signalering, binnen het onderwijs (tijdig) op geschakeld kan worden. Vanuit het Deltion wordt benoemd dat op school met name de logistieke context
wordt geleerd en dat de breedte en details worden aangeleerd in de praktijk. Hieraan
zijn verschillende (praktijk) opdrachten gekoppeld om de student bewust te maken
van zijn kennis en de mogelijkheden die er in de praktijk zijn. Hierin speelt ook de
ervaring van de (praktijk)begeleider in de begeleiding. Hoe ervarener de begeleider,
hoe meer de student in staat gesteld wordt om veel (verschillende) logistieke
onderdelen van een organisatie te zien én te ervaren. Vanuit het STC wordt hierbij aangevuld dat het werken met praktijkproblemen (vanuit
concrete praktijkvraagstukken van bedrijven) ook een mooie manier is om juist met de
praktijk in aanraking te komen en te leren. Het is van belang om het doel van de
opdracht goed door te spreken met het bedrijf om ook een concreet leerdoel te
kunnen nastreven. Hierin speelt veelvuldig een trial-and-error aanpak. Vooraf
uitdenken en achteraf evalueren of hetgeen dat was uitgedacht ook is behaald. www.sharehouselab.nl Rol van Living Lab Voor een structurele samenwerking met het bedrijfsleven in een living lab omgeving
zoals het Sharehouse en Logistics Lab zou een business case gemaakt kunnen worden
waarbij een win-win situatie tussen opleiding en bedrijf centraal staat. Hierbij kan op
verschillende manieren gebruik gemaakt worden van de fysieke omgeving. Denk
bijvoorbeeld aan: contractonderwijs voor bijscholing van medewerkers, het uitvoeren
van experimenten, student challenges etc. Belangrijk is dat zowel het bedrijfsleven als
het onderwijs hier een meerwaarde inzien en er commitment aan geven. www.sharehouselab.nl Visie HBO Ook binnen het HBO wordt de rol van het bedrijfsleven steeds belangrijker aangezien
praktijkprojecten, buiten stage en afstuderen, steeds centraler staan in het
opleidingsprogramma. Een structurele en strategische samenwerking met een aantal
bedrijven lijkt steeds belangrijker te worden om de kwaliteit en begeleiding van de
projecten te kunnen waarborgen. www.sharehouselab.nl LANDELIJK ONDERWIJS In de voorgaande onderdelen zijn verschillende aandachtsgebieden benoemd alsook
verschillende initiatieven die vanuit de verschillende MBO en HBO instellingen zijn
en/of worden ondernomen om in te spelen op deze aandachtsgebieden. Tegelijkertijd
reist de vraag, als deze aandachtsgebieden onvoldoende landelijke aandacht krijgen
blijven het losstaande initiatieven waarin in iedere regio eigen initiatieven en
onderzoeken worden ontplooid. Het doel van is enerzijds om inzicht te geven in het
huidige onderwijs vanuit de deelnemende onderwijsinstellingen en anderzijds
handvatten te geven voor de gemeenschappelijke opleidingsprofielen op landelijk
niveau. Deze stap wordt in dit onderdeel verder benoemt. Zowel binnen het MBO als het HBO wordt gebruik gemaakt van een gemeenschappelijk
profiel voor alle logistieke opleidingen. Vanuit het MBO wordt dit gestuurd vanuit het
SBB Sectorkamer (Samenwerkingsorganisatie Beroepsonderwijs Bedrijfsleven), waarbij
binnen het HBO vanuit het LPL (Landelijk Platform Logistiek) een gezamenlijke visie is
ontwikkeld. Beiden zullen hierna kort en individueel worden behandeld. 21 Wat opvalt is dat in beide keuzedelen kennis en informatie rondom stuurinformatie
zowel in het lezen als het opstellen ervan niet worden benoemd. Als we naar de
landelijke kwalificatiedossiers kijken, zien we hierin ook dat het werken met of aan de
hand van stuurinformatie nauwelijks wordt benoemd. Op MBO 4 niveau wordt het
benoemd in het kader van personeelsbeleid, maar verder komt dit niet terug. Wat opvalt aan de hand van de keuzedelen, als ook bij het doornemen van de
kwalificatie dossiers is dat gedragsvormen en sociaal handelen overal terugkomt. Echter een kritische blik en daaraan gelieerde houding en vaardigheden in een
‘samenwerkende’ omgeving komt nergens echt concreet naar voren. Op basis van de
resulaten van ons onderzoek zou dat meer aandacht behoeven bij de verschillende
logistieke opleidingen. Tevens is het onderwerp waarin de MBO-student (onafhankelijk van het MBO niveau)
wordt geconfronteerd met het ontwikkelen van zijn eigen vaardigen in een sterk
veranderende omgeving te beperkt aanwezig. Het gaat over zaken als wendbaarheid en
flexibiliteit; hoe ga jij als toekomstige medewerker om met elke dag andere instructies,
hoe sta jij stil bij innovaties en ben jij in staat hierin de mogelijkheden te zien zijn
onderwerpen die vanuit de resultaten van het onderzoek meer inhoud kan geven aan
de MBO professional van de toekomst. Vanuit het HBO onderwijs geldt dat bij het schrijven van deze rapportage (begin 2023)
het ‘Beroeps- en opleidingen profiel Logistiek’ uit 2019 (Vries, Crean, & Mandemakers,
2019) als basis is genomen voor een korte reflectie. In het document ‘Beroeps- en opleidingen profiel Logistiek’ wordt uiteindelijk
onderscheid gemaakt tussen hard en soft skills. Tabel nr xxx: Hard en soft skills (LPL, 2019) MBO-onderwijs Vanuit het MBO-onderwijs wordt vanaf augustus 2022 gewerkte met nieuwe
kwalificatiedossiers voor de logistieke opleidingen. Met name onderwerpen als ICT en
digitale vaardigheden zijn hierin sterker aangezet dan in de voorgaande
kwalificatiedossiers (Transport & Logistiek, 2022). Hierin wordt benadrukt dat er
keuzedelen zijn toegevoegd voor alle MBO niveaus. Zo is er een keuzedeel ‘Digitale
vaardigheden basis’ opgesteld met name voor MBO 2 en MBO 3 niveau en is er een
keuzedeel ‘Digitale vaardigheden gevorderd’ opgesteld voor MBO 3 en MBO 4 niveau. Deze keuzes worden aangeboden naast het in 2017 ontwikkelde keuzedeel
‘Automatisering met behulp van ERP-systemen’. In de nieuw ontwikkelde keuzedelen
ligt er vanuit de kennis & vaardigheden veel focus op aspecten waarop gelet moet
worden wanneer gewerkt wordt met data (basis) en is men in staat om data
zelfstandig te verwerken en vast te leggen (gevorderd). Met name binnen het
keuzedeel ‘automatisering met behulp van ERP-systemen’ wordt meer connectie
gemaakt met ICT systemen die binnen de logistieke context gebruikt worden, waarbij
ook koppelingen worden gemaakt met stamdata, verwerken van gegevens maar ook
kennis van innovatieve toepassing van hardware in productie en logistieke
omgevingen. www.sharehouselab.nl 21 Hard skills De regie- en architectenfunctie van logistiek
doet zich niet alleen conceptueel en abstract
voor. De sociale context wordt belangrijker:
klantcontacten, interne en externe
samenwerking, managen van
veranderingsprocessen etc. Om daadwerkelijk
bedrijfsprocessen op elkaar af te stemmen en
als één geheel te laten functioneren, dient de
logisticus over sociale en communicatieve
vaardigheden te beschikken. Motiveren, coachen
en kunnen onderhandelen verdienen hierbij veel
aandacht27. Logistiek is mensenwerk: met
mensen en voor mensen. Om in dit werkveld te
kunnen opereren/samenwerken en
daadwerkelijk veranderingen te realiseren zijn de
soft skills van fundamenteel belang. Dit betreft voor een deel gereedschappen die
ook snel veranderen. Het is daarom belangrijk
dat de logisticus over analytische vaardigheden
beschikt, waarmee hij/zij de bedrijfsprocessen
kan doorgronden en ICT-tools optimaal benut. Daarnaast worden ook wiskundige skills
belangrijker (met name vereist op wo-niveau). De behoefte aan digitale vaardigheden
in de logistiek zijn van fundamenteel belang. www.sharehouselab.nl 22 Aan de hand van dit onderscheid zal het ‘Beroeps- en opleidingen profiel Logistiek’ ook
worden bekeken. HARD skills In het document ‘Beroeps- en opleidingen profiel Logistiek’ (Vries, Crean, & In het document ‘Beroeps- en opleidingen profiel Logistiek’ (Vries, Crean, &
Mandemakers, 2019) worden verschillende innovaties benoemd alsook het kunnen
vertalen van de innovaties naar concrete bedrijfstoepassingen. De voorbeelden die
hierin met name worden behandeld, lijken wel sterker gericht op de fysieke innovatie
en minder op de digitale innovatie. Eén van de leerpunten vanuit het onderzoek naar
de veranderende skills in een warehousecontext (Grolleman, Hendrickx, Warffemius,
Vos, & Versluis, 2022) was met name het meer en concreter kunnen omgaan met het
vertalen van data naar dashboards en stuurinformatie. Zoals ook al eerder is benoemd,
zijn de Logistics Engineering opleidingen van Hogeschool Rotterdam en Fontys Venlo
concreet mee bezig om dit in het onderwijs te verankeren. Vanuit dit onderzoek zou onze aanbeveling zijn om dashboards, stuurinformatie en
daaraan gekoppelde BI-tooling (zoals PowerBI, Tableau of QlickSense) concreet te
benoemen in de volgende versie van het ‘Beroeps- en opleidingen profiel Logistiek’
zodra het weer opnieuw wordt ontwikkeld. Tevens is eerder benoemd dat, met name de Logistics Engineering opleidingen, het
programmeren van codes ter verwerken van ‘big data’ (middels Python en/of R) aan
het verkennen zijn of al hebben opgenomen in hun curricula. Aanbeveling is dit ook op
te gaan nemen in de komende versie van het ‘Beroeps- en opleidingen profiel
Logistiek’. www.sharehouselab.nl SOFT skills Soft skills worden vanuit het ‘Beroeps- en opleidingen profiel Logistiek’ (Vries, Crean, &
Mandemakers, 2019) op verschillende plekken concreet benoemd, alsook het belang
hiervan. Tegelijkertijd wordt ook aangegeven dat op “een aantal human capital thema’s
nog onderzoek is te doen. De belangrijkste hiervan is sociale innovatie. Hiermee wordt
bedoeld de herinrichting van onderwijs, arbeid en arbeidsmarkt voor de logistiek,
waardoor werkgeverschap en werknemerschap naar een hoger kwaliteitsniveau
worden getild. Onderzoek moet zich met name concentreren op de relatie tussen
organisatievormen bij logistieke bedrijven en hun betrokkenheid bij en adoptie van
innovatie”. Dit wordt verderop in het ‘Beroeps- en opleidingen profiel Logistiek’
vertaald naar “Logistici zullen over innovatieve skills moeten beschikken én bereid zijn
tot een leven lang leren om in te kunnen spelen op de snelle veranderingen binnen (en
in relatie tot) de logistiek” Uiteindelijk resulteert dit in een opsomming van professioneel vakmanschap
waaronder ook een wendbare professional hoort. Hieronder wordt verstaan: “De
afgestudeerde hbo-professional kan proactief inspelen op veranderingen in het werk
en de context van het werk. Hij of zij kan omgaan met weinig structuur en onzekere
factoren: ‘knowing what to do, when you don’t know what to do.’ Hij of zij ontwikkelt
hiertoe voortdurend de eigen professionaliteit en draagt bij aan de ontwikkeling van www.sharehouselab.nl 23 zijn of haar professie in de breedte. Daarbij is de hbo-professional in staat doelen voor
de korte termijn te verbinden met ambities voor de lange termijn en houdt daarbij oog
voor de snel veranderende omgeving en de grote hoeveelheden beschikbare
informatie (big data)” (Vries, Crean, & Mandemakers, 2019). Naar aanleiding van deze inzichten, lijkt een 1-op-1 relatie te liggen met de resultaten
van het onderzoek naar de veranderende skills in een warehouse-context (Grolleman,
Hendrickx, Warffemius, Vos, & Versluis, 2022). Echter wanneer het vervolg van het
‘Beroeps- en opleidingen profiel Logistiek’ wordt doorgenomen wordt er een brug
gemaakt naar een concreet verbetervoorstel (vanuit LM-perspectief) of een concreet
ontwerp (vanuit Leng-perspectief). Wanneer vervolgens een vertaling van het geheel
wordt gemaakt naar de logistieke deelcompetenties (als verantwoording voor de
logistieke HBO opleidingen) komt de focus meer te liggen op het veranderen in de
context van een doel, waarbij de huidige technologische ontwikkelingen wellicht meer
innovativiteit en wendbaarheid van zowel de professional als de organisatie vragen. Hierbij lijkt het begrip van ‘sociale innovatie’ zoals ook eerder is benoemd weer sterker
naar voren te komen. SOFT skills Sterker nog, er wordt in het ‘Beroeps- en opleidingen profiel Logistiek’ gesproken over
persoonlijk leiderschap (wat een vertaling is van stil kunnen staan bij de eigen
ontwikkeling) als voorwaarden van doorgroei naar leidinggevende functies. Dit roept de
vraag op, of doorgroei naar een leidinggevende functie de énige doorgroeimogelijkheid
is. Als vanuit dit perspectief naar de gedefinieerde BoKS (Body of Knowledge and Skills)
binnen het ‘Beroeps- en opleidingen profiel Logistiek’ wordt gekeken komen hoogover
de begrippen ‘organizational behavior’ en ‘communication skills’ naar voren. Dieper
kijken naar de begrippen en inhoud die hierin wordt benoemd, adviseren wij om binnen
‘organizational behavior’ ook de term sociale innovatie als onderwerp op te nemen
binnen de BoKS voor zowel de Leng als de LM opleiding. Logischerwijs is hierin qua
diepgang en aanpak een onderscheid te adviseren naar Leng en LM opleidingen. Tevens zouden we de aanbeveling willen doen om binnen ‘communication Skills’ ook
de professionele wendbaarheid en innovatieve mindset toe te voegen als skills
waaraan iedere Logistieke opleiding handen en voeten dient te geven. www.sharehouselab.nl Conclusie Binnen deze rapportage is inzicht gegeven in de belangrijkste resultaten vanuit het
onderzoek naar de veranderende skills door toepassing van meer ICT technologie in
een warehousecontext en de impact daarvan op het logistieke MBO en HBO onderwijs. Er is inzicht gegeven in zowel een aantal ontwikkelingen binnen het MBO en HBO
onderwijs vanuit de deelnemende kennisinstellingen; STC, Deltion, HR, Fontys Venlo,
Windesheim. Hieronder zullen enkele conclusies worden benoemt die in het vervolg
van onderwijsontwikkelingen in het logistieke domein aandacht verdienen. www.sharehouselab.nl 24 Het combineren van bedrijfsleven en onderwijs door middel van een living lab zoals het
Sharehouse is een waardevolle manier om contextrijk onderwijs te bieden waarin (alle)
studenten kunnen leren van zo concreet mogelijke praktijksituaties, zonder gebonden
te zijn aan de (individuele) begeleiding van één specifieke docent. Tevens bieden living labs een mooie oefen (en proef-) tuin om de verschillende
onderwijsniveaus van MBO 2 tot HBO niveau met elkaar in aanraking te laten komen en
daarmee ook van elkaar te leren. Hierbij kan gedacht worden aan initiatieven als het
warehouse van de toekomst en Deltion E-Logistics waarin MBO en HBO docenten op
verschillende manier met andere doelen in één project samenwerken. Het bedrijfsleven dient een (grotere) rol te gaan spelen in het ontwerpen en bij
voorkeur actief te participeren in het onderwijs waarbij het doel moet zijn om het
onderwijs zo gelijk mogelijk voor iedere student aan te bieden en het leereffect minder
afhankelijk is van een individuele praktijkbegeleider. Hierbij kan gedacht worden aan de
inzet van het Soft Skills Lab waarin bedrijven concrete praktijkvraagstukken aandragen
met als doel (MBO) studenten zelf te laten ervaren wat hun eigen houding en gedrag is
in verschillende situaties. Vanuit de huidige kwalificatiedossier op MBO niveau, lijkt het onderwerp rondom
digitalisering en data nog sterk gefocust te zijn rondom daadwerkelijk uit te voeren
taken & handelingen en minder op de wijze waarop digitalisering kan bijdragen en wat
de impact van digitalisering gaat zijn op het handelen van de MBO student. Daarnaast
is het advies om meer stil te staan bij reflectie met de MBO studenten in het
ontwikkelen van eigen leeractiviteiten aan de hand van technologische innovaties. Binnen het HBO niveau zien we op zowel Leng als LM opleidingen mooie (individuele)
initiatieven die in lijn liggen met de veranderende skills. De wijze waarop deze
veranderingen terug te vinden zijn in het huidige landelijke ‘Beroeps- en opleidingen
profiel Logistiek’ is nog te weinig concreet. Conclusie Hiervoor zijn een aantal aanbevelingen
benoemd om dit verder te complementeren en als Logistieke HBO-opleidingen met
elkaar na te streven en bij voorkeur gezamenlijk op te trekken. Hierbij kan gedacht
worden aan het integreren van begrippen als BI-tooling, programmeren voor beginners
en sociale innovatie vanuit een kennismindset. Geredeneerd vanuit de houding en
gedrag van studenten zouden professionele wendbaarheid en innovatieve mindset als
waardevolle toevoegingen worden geadviseerd om op te nemen. Nagedacht dient te worden hoe vanuit LM-perspectief naar Leng onderwerpen (zoals
Dashboarding & programmeren) te kijken en vanuit Leng-perspectief naar een
onderwerp als sociale innovatie. Het lijkt er daarbij op dat de onderwijsgebieden van
Leng en LM minder afzonderlijk zouden kunnen zijn dan momenteel is ingericht, maar
vooral afhankelijk is van de mindset van de professional. In essentie dienen, vanuit de
resultaten van dit onderzoek, de LM studenten meer technologische hard skills te
ontwikkelen en Leng studenten juist meer soft skills. www.sharehouselab.nl www.sharehouselab.nl Kritische reflectie Dit onderzoek is uitgevoerd aan de hand van de resultaten uit de Logistics Skills of the
Future Scan die is ontwikkeld (Grolleman, Hendrickx, Warffemius, Vos, & Versluis,
2022), waarbij verschillende gesprekken zijn gevoerd met opleidingsmanagers en
curriculum commissies van het STC, Deltion, Hogeschool Rotterdam, Fontys
Hogescholen Venlo & Hogeschool Windesheim. Belangrijk om te vermelden is dat het onderzoek heeft plaats gevonden met een
afvaardiging van een beperkt aantal MBO- en HBO-opleidingen. Deze rapportage is, als
eindresultaat, ook door (een deel van) de geïnterviewden onderschreven. Vanwege
beschikbare tijd is niet van iedere geïnterviewde een schriftelijke reactie gekomen. Hierdoor is de generaliseerbaarheid van dit onderzoek te betwisten, waarmee deze
rapportage vooral als inspiratie dient te worden gezien voor landelijke opleidingskaders
in zowel het logistieke MBO als HBO onderwijs. www.sharehouselab.nl 25 Tenslotte dienen de succesvolle pilots m.b.t. samenwerking tussen MBO en HBO
voortgezet worden. Denk bijvoorbeeld aan een onderzoek dat heeft plaatsgevonden
vanuit het Zwolse initiatief Logistics Lab mbt de Exoskelets. www.sharehouselab.nl www.sharehouselab.nl 26 De beoogde methodiek van deze Learning Community gaat uit van de bestaande
beroepsfuncties en door de competenties en vaardigheden in een beroepsfunctie te
vergelijken met de potentie van nieuwe technologie of eisen van de business
omstandigheden kan onderbouwd komt de verwachte verandering beter in beeld. De
tweede stap is het uitwerken van het transitiepad in termen van nieuwe functies, het
binden, leven-lang-leren en ontwikkelen van huidige medewerkers en het aantrekken
van nieuwe medewerkers. Binnen de Learning Community is ook een skill-vacature
monitor ontwikkeld waarmee de sector brede ontwikkelingen in gevraagd skills kunnen
worden gevolgd. In 2023 is gestart met 12 bedrijven in Oost-Nederland (Gelderland,
Overijssel) en Rotterdam. De learning community is open en bedrijven die een
vraagstuk willen oppakken en hun vragen, kennis en ervaring actief willen delen kunnen
aansluiten. De kennis en inzichten die in Sharehouse zijn ontwikkeld over de ontwikkeling van skills
in het warehouse van de toekomst worden meegenomen in de skills-vacature monitor. De Learning Community Future Skills in Logistics en Sharehouse zijn complementaire
initiatieven en kunnen bij een eventueel vervolg van Sharehouse de samenwerking
zoeken als er bedrijven in de Learning Community zijn die specifiek met skills
vraagstukken in warehousing aan de slag willen. Vanuit Sharehouse kunnen de al
geïdentificeerde skills verder worden aangescherpt. In de Learning Community zal
aandacht zijn voor de wijze waarop het bedrijf de verwachte of gewenste
veranderingen kan identificeren en kan omzetten naar maatregelen. Aanbevelingen voor Verder onderzoek Uiteraard stopt ook het leren naar de toekomstige skills niet. Vanuit het Sharehouse-
project, specifiek vanuit het onderdeel Skills of the Future in Education, is een focus
gelegd op de toekomstige skills in een veranderende warehousecontext. Daarmee zijn
de resultaten van dit onderzoek sterk gefocust op een warehouse omgeving en is
verder onderzoek noodzakelijk om het geheel ook breder te trekken naar de gehele
logistieke sector. Uiteraard zijn door de verschillende gesprekken in zowel de validatie van de resultaten
als ook het vaststellen van het onderwijs (zowel op MBO als HBO niveau) gedeeld met
de verschillende Curriculum Commissies met als doel de uitkomsten zo breed mogelijk
te kunnen toepassen. Het is ook mooi te zien dat ook de Logistieke sector erkent dat zij nog niet klaar is en
wil blijven leren en ontwikkelen met betrekking tot de toekomstige skills van logistieke
medewerkers op verschillende niveaus in bedrijven. Zo is er, sinds medio 2022, een
onderzoek gaande waarin een Learning Community Future Skills in Logistics is gestart
om zicht kunnen krijgen op skills die in de toekomst nodig zijn en hoe bedrijven hun
personeel en het personeelsbestand daarop kan voorbereiden. In de LC Future Skills in
Logistics wordt dit vraagstuk bottum-up vanuit cases van de deelnemende bedrijven
verkend en ontwikkelen de kennisinstellingen aan de hand van de cases een
methodiek die bedrijven en medewerkers kunnen volgen om meer inzicht te krijgen De
Learning Community Future Skills in Logistics is een samenwerking van Hogeschool
Rotterdam, Hogeschool Arnhem Nijmegen, Windesheim, TLN en Evofenedex. www.sharehouselab.nl 26 www.sharehouselab.nl 27 27 Bibliografie
Grolleman, H., Hendrickx, R., Warffemius, P., Vos, M., & Versluis, P. (2022). De
belangrijkste logistieke logistieke skills in een veranderende warehousecontext. Logistiek+, pp. 98-123. Transport & Logistiek. (2022, april 7). Meer aandacht voor ICT in mbo-opleidingen
logistiek. Transport & Logistiek. Vries, R. d., Crean, J. d., & Mandemakers, A. (2019, 11 22). Beroeps- en opleidingsprofiel
Logistiek - ten behoeve van de bacheropleidingen Logistics Management &
Logistics Engineering. Opgehaald van Vereniging hogescholen:
https://www.vereniginghogescholen.nl/system/profiles/documents/000/000/2
45/original/LPL_Beroeps-_en_Opleidings_Profiel_2019.pdf?1580199559 Grolleman, H., Hendrickx, R., Warffemius, P., Vos, M., & Versluis, P. (2022). De
belangrijkste logistieke logistieke skills in een veranderende warehousecontext. Logistiek+, pp. 98-123. Transport & Logistiek. (2022, april 7). Meer aandacht voor ICT in mbo-opleidingen
logistiek. Transport & Logistiek. Vries, R. d., Crean, J. d., & Mandemakers, A. (2019, 11 22). Beroeps- en opleidingsprofiel
Logistiek - ten behoeve van de bacheropleidingen Logistics Management &
Logistics Engineering. Opgehaald van Vereniging hogescholen:
https://www.vereniginghogescholen.nl/system/profiles/documents/000/000/2
45/original/LPL_Beroeps-_en_Opleidings_Profiel_2019.pdf?1580199559 Vries, R. d., Crean, J. d., & Mandemakers, A. (2019, 11 22). Beroeps- en opleidingsprofiel
Logistiek - ten behoeve van de bacheropleidingen Logistics Management &
Logistics Engineering. Opgehaald van Vereniging hogescholen:
https://www.vereniginghogescholen.nl/system/profiles/documents/000/000/2
45/original/LPL_Beroeps-_en_Opleidings_Profiel_2019.pdf?1580199559 Bijlage 01 – Resultatentabel Logistics Skills of the Future ( Bijlage 01 – Resultatentabel Logistics Skills of the Future ( Bijlage 01 – Resultatentabel Logistics Skills of the Future ( Bijlage 01 – Resultatentabel Logistics Skills of the Future (
Voor meer informatie en toelichting, zie ook (Grolleman, Hendrickx, Warffemius, Vos, &
Versluis, 2022). Voor meer informatie en toelichting, zie ook (Grolleman, Hendrickx, Warffemius, Vos, &
Versluis, 2022). -
Zowel het bedrijfsleven als het onderwijs moeten een win-win situatie hebben www.sharehouselab.nl www.sharehouselab.nl Omschrijving
HBO
R’da
m
HBO
Venlo
HBO
Zwoll
e
MBO
R’da
m
MBO
Zwoll
e
Data science module (bijlage 3 en 4)
X
X
nvt
nvt
Warehouse van de toekomst in relatie tot
trends in living lab (bijlage 3)
X
Management Dashboards (bijlage 3 en 4) in
power BI en Tableau
X
X
Onderzoek toepassing EXO skelets in
warehouse omgeving in living lab (pagina 5)
X
X
Sharehouse lab
-
Pick-to-light car
-
een scan systeem voor orderpicking
-
Exact online WMS pakket
-
Vision Picking door gebruik van een head
mounted display
X
X
X
X
Living Lab
-
een Pick-to-light stelling
-
een scan systeem voor orderpicking
-
een eigen ingericht ERP & WMS systeem
-
een dozen inpak machine
X
X
X
X
Voorbeelden vanuit opleidingen op het gebied van hard skills
Soft skills
Omschrijving
HBO
R’da
m
HBO
Venlo
HBO
Zwoll
e
MBO
R’da
m
MBO
Zwoll
e
Training in soft skills lab – communicatieve
vaardigheden – (bijlage 5)
X
E-logistics – Casestudy procesverbetering
(bijlage 6) en lean orange belt (bijlage 7)
X
X
Sociale innovatie binnen een lerende
organisatie
(bijlage 8)
X
Omgaan met weerstanden en draagvlak creëren
(bijlage 9)
X
Voorbeelden vanuit opleidingen op het gebied van soft skills. www.sharehouselab.nl www.sharehouselab.nl Bijlage 03 – Data Science in het HBO-onderwijs (Fontys)
Opzet Data Science Fontys – Introductie Data Science en management dashboards Bijlage 03 – Data Science in het HBO-onderwijs (Fontys) Context: Projectmatig werken Projectmatig werken komt in elk onderwijsblok terug. Dit kan zijn in de vorm
van een fictief project of een praktijk project. In dit blok is het nog een fictief
project wegens gebrek aan data van een bedrijf om een dashboard op te
bouwen. www.sharehouselab.nl Context: Ontwerpen van een Management Dashboard Elk logistiek bedrijf beschikt over grote hoeveelheden gegevens uit
verschillende informatiesystemen. Met gegevens alleen kunnen we niets, deze
gegevens moeten worden omgezet in bruikbare management- en/of
stuurinformatie. Een manier om deze informatie overzichtelijk te presenteren is
het opzetten van een management dashboard. Voordat een management dashboard wordt opgezet, dient men eerst inzicht te
krijgen in de strategie van de organisatie, de daarbij behorende kritische
succesfactoren en de daarvan afgeleide KPI’s per deelgebied. Deze
deelgebieden moeten in balans zijn en kunnen worden weergegeven in een
balanced scorecard (BSC). De logistiek verantwoordelijke moet inzicht hebben
in alle deelgebieden, dat wil zeggen dat hij/zij logistiek niet alleen vanuit een
bedrijfsprocesperspectief moet bekijken, maar ook vanuit een financieel
perspectief, een klantenperspectief en een innovatieperspectief. Na het
formuleren van relevante kengetallen en KPI’s wordt er in PowerBi een
datamodel opgezet en relaties gelegd tussen de verschillende kengetallen. Uiteindelijk worden de kengetallen gevisualiseerd in een dashboard. Een veelgehoorde term met betrekking tot het maken van management
dashboards is de toepassing van Data Science methodieken en tools. Dit is een
georganiseerd en systematisch proces waarbij organisaties gegevens uit zowel
interne als externe informatiebronnen verkrijgen, analyseren en verspreiden om
hun bedrijfsvoering en besluitvorming te kunnen ondersteunen. De studenten
leren in dit semester de eerste stappen te zetten in R. R is een softwarepakket
en een programmeertaal ontwikkeld voor statistiek en gegevensanalyse. Logistics Engineers zullen in hun beroep regelmatig deel uitmaken van een team
om een dashboard op te zetten of aan te passen en statistische analyses door
te voeren. -
Zowel het bedrijfsleven als het onderwijs moeten een win-win situatie hebben www.sharehouselab.nl -
Is er een business case te maken? -
Welke bedrijven kunnen een set data aanleveren in een warehouse context? -
Welke toegevoegde waarde heeft voor het bedrijf? Doel: een dashboard dat ook door de
deelnemende bedrijven geïmplementeerd wordt of gesimuleerd wordt. -
Als dit business model succesvol is, kan worden bekeken hoe MBO en HBO studenten
binnen het businessmodel ook kunnen leren en experimenteren in het Sharehouse. -
Is er een business case te maken? -
Als dit business model succesvol is, kan worden bekeken hoe MBO en HBO studenten
binnen het businessmodel ook kunnen leren en experimenteren in het Sharehouse. Context: Relevantie Data Science Wanneer men naar een keynote luistert of een industrieel evenement bezoekt,
wordt men overspoeld met modewoorden als big data, cloud computing,
kunstmatige intelligentie oftewel AI, blockchain, eerlijke en verantwoorde
logistiek, sharing economie en nog veel meer. Deze nieuwe technologieën en
logistieke concepten hebben de mogelijkheid om de huidige praktijken in de
toeleveringsketen volledig te hervormen en kunnen een grote impact hebben
op de manier waarop processen er in de toekomst uit zullen zien. Twee van deze ontwikkelingen, blockchain en kunstmatige intelligentie,
vertonen een groot potentieel om binnenkort in organisaties toe te passen. In
deze CAS zullen studenten kennismaken met een breed scala aan van
onderwerpen en zullen ze proberen nieuwe ontwikkelingen in de logistieke
praktijk toe te passen. Wanneer men naar een keynote luistert of een industrieel evenement bezoekt,
wordt men overspoeld met modewoorden als big data, cloud computing,
kunstmatige intelligentie oftewel AI, blockchain, eerlijke en verantwoorde
logistiek, sharing economie en nog veel meer. Deze nieuwe technologieën en
logistieke concepten hebben de mogelijkheid om de huidige praktijken in de
toeleveringsketen volledig te hervormen en kunnen een grote impact hebben
op de manier waarop processen er in de toekomst uit zullen zien. T
d
t ikk li
bl
k h i
k
t
ti
i t lli
ti De leerdoelen geven aan wat studenten aan het eind van de leerlijn moeten
beheersen De student weet welke nieuwe technologieën en logistieke
concepten een rol zullen spelen in de toekomstige toeleveringsketen en kan
een analyse maken in hoeverre hun toepassing ervan kan bijdragen aan het
verbeteren van processen en het bedrijfsresultaat. Hij/zij kan ook de mogelijke
toepassing van deze ontwikkelingen en nieuwe technologieën op een (fictieve)
business case en kan suggesties doen in zijn/haar toekomstige werksituatie om
hiervan gebruik te maken. De volgende onderwerpen zullen aan de orde komen binnen nieuwe De volgende onderwerpen zullen aan de orde komen binnen nieuwe
ontwikkelingen: De volgende onderwerpen zullen aan de orde komen binnen nieuwe
ontwikkelingen: • Inleiding: Supply chain and technology trends (waaronder robotics in
warehouses) • Belangrijkste aspecten van AI • Blockchain en mogelijke toepassingen • AI, Blockchain en Robotics proces automatiseringen Context: de opdracht Studenten wordt gevraagd hun vaardigheden in het toepassen van deze nieuwe
technologieën op processen aan te tonen en bewijs te leveren van de
bruikbaarheid (uit literatuur en empirische studies). Empirisch bewijs kan ook
worden verzameld via interviews met relevante belanghebbenden die deze
technologieën of concepten hebben geïmplementeerd. De volgende onderwerpen komen in deze data science cursus aan de orde g
p
• De CRSIP DM model
• R part 2
• Time series forecasting
• Forecasting with Knime
• Forecasting with R
• Data mining
• Proces mining
• Simulation • De CRSIP DM model
• R part 2
• Time series forecasting
• Forecasting with Knime
• Forecasting with R
• Data mining
• Proces mining
• Simulation p
1. Toepassen van forecasting met datasets uit de praktijk 2. Toepassen van simulatie met behulp van aangeleerde programma’s De activiteiten krijgen vorm door middel van individuele opdrachten en
samenvoegen daarvan in een portfolio. De activiteiten krijgen vorm door middel van individuele opdrachten en
samenvoegen daarvan in een portfolio. www.sharehouselab.nl www.sharehouselab.nl Bijlage 04 – Data Science in het HBO onderwijs (Hogeschool
Rotterdam) Bijlage 04 – Data Science in het HBO onderwijs (Hogeschool
Rotterdam) Opzet Ontwerpen van een toekomstbestendig warehouse Ongeveer een kwart van de opleiding Leng bestaat uit projectonderwijs. Kenmerkend voor onderwijsprojecten is dat de student leert zelfstandig als
professional te werken. In het project zal de student de kennis die hij heeft
opgedaan in de gerelateerde vakken verdiepen, aanvullen en toepassen op een
(fictieve) situatie uit de praktijk. Niet alleen op school wordt projectmatig gewerkt, ook in het bedrijfsleven
wordt steeds meer op deze manier gewerkt. Het bedrijfsleven ondergaat vele
invloeden. Invloeden van de zijde van afnemers, aandeelhouders, overheid en
niet in het minst van concurrenten. Afnemers veranderen continu hun wensen
en voorkeuren, concurrenten ontwikkelen continu nieuwe producten, overheid,
medewerkers en aandeelhouders schroeven hun eisen op. Dit leidt ertoe dat
ondernemingen hun aanpak steeds dienen aan te passen aan de veranderde
omgeving. Deze zaken worden in multidisciplinaire teams (projectteams)
opgelost. Het is van belang dat de student leert omgaan met projectonderwijs en
projectmatig werken. Op school en natuurlijk in zijn latere werk als logisticus. Het werken in projecten is iets dat de student moet leren. Een project is iets
anders dan het doen van een tentamen: de student moet samenwerken met
medestudenten, hij moet zelfstandig problemen kunnen oplossen, hij moet
vakoverstijgend kunnen denken en hij moet de juiste informatie weten te
vinden en toepassen. De kennis en ervaring die de student al heeft opgedaan
over het projectmatig werken heeft hij weer nodig in dit project en gaat hij
verder verdiepen. www.sharehouselab.nl www.sharehouselab.nl Context: Ontwerpen van een Warehouse Het ontwerp van het warehouse bepaalt in grote mate hoe goed het
functioneert. In een goed ontworpen warehouse komen fouten minder vaak
voor, wordt het beheer van de voorraden slim aangepakt, kunnen de voorraden
vaak lager zijn, gaat het orderpicken sneller, is de veiligheid beter (minder
ongelukken), is beter nagedacht over milieu en duurzaamheid, is de
informatiestroom beter op orde, is de service aan de klant hoger en zijn de
kosten lager. Logistics Engineers zullen in hun beroep regelmatig deel uitmaken
van een team om een warehouse opnieuw te ontwerpen of een bestaand
ontwerp te optimaliseren. www.sharehouselab.nl Context: Ontwerpen van een Toekomstbestendig Warehouse De logistieke sector maakt grote ontwikkelingen en innovaties door. Denk aan
alle ontwikkelingen op het gebied van ICT zoals: the Internet of Things (IoT),
sensor technologie, robots, virtual reality (VR), augmented reality (AR), of big
data. Logistics Engineers zullen steeds vaker komen te werken in een digitale
omgeving. In de logistieke keten van de toekomst zal het steeds meer om flexibiliteit,
snelheid en duurzaamheid draaien. Die keten – en dus ook het warehouse- zal
moeten transformeren om de veranderende marktvraag aan te kunnen. Het
gaat dus om de vraag hoe het warehouse toekomstbestendig te maken zodat
het mee kan in de logistieke ontwikkelingen van de toekomst. En technologie
helpt daarbij. In een toekomstbestendig warehouse zijn de goederen- en
informatiestroom met elkaar verbonden. Denk bijvoorbeeld aan: de
administratie van spullen bij levering (binnenkomst), coördinatie van goederen
die een plek krijgen in het magazijn, het werken met automatische
bestelniveaus van de voorraden in het warehouse, coördinatie van het
orderpicken of de aansturing van de distributie van goederen (bij welke klant
moet welk product wanneer worden afgeleverd). Het warehouse van de 21e
eeuw is een high-tech hal waarin bijna alles is geautomatiseerd. De relatie van deze module met de kerncompetenties van Leng staan in
onderstaand schema. Tabel 1: Leerdoelen en competenties www.sharehouselab.nl www.sharehouselab.nl www.sharehouselab.nl www.sharehouselab.nl h
h
l b l www.sharehouselab.nl www sharehouselab nl Directe koppeling met Sharehouse: lessen in Sharehouse organiseren Afgelopen cursusjaar hebben twee lessen in het Sharehouse plaatsgevonden. Dat is echt een grote meerwaarde gebleken. Dit zouden wij samen met
Sharehouse verder vorm willen geven en structureren voor komende
cursusjaren. -
Is er een business case te maken? -
Wat gemaakt kan worden is een logistiek business plan. Breng de waarde proposities in
kaart ahv het business model canvas. -
Wat gemaakt kan worden is een logistiek business plan. Breng de waarde proposities in
kaart ahv het business model canvas. -
Denk bijvoorbeeld aan het maken van een trainingsprogramma ism het bedrijfsleven. Een trainingsprogramma in het Sharehouse voor medewerkers van het bedrijfsleven in
de vorm van contractonderwijs. Sharehouse wordt dan gepositioneerd als een
bedrijfsschool voor een aantal bedrijven (gedeelde opleidingscapaciteit). -
Start met het inventariseren wat het bedrijfsleven aan scholing nodig heeft voor haar
eigen medewerkers en wat het Sharehouse daarin kan bieden. -
Als dit business model succesvol is, kan worden bekeken hoe MBO en HBO studenten
binnen het businessmodel ook kunnen leren en experimenteren in het Sharehouse. www.sharehouselab.nl www.sharehouselab.nl www.sharehouselab.nl www.sharehouselab.nl Bijlage 05 - Opzet Data Science for logistici (Hogeschool
Rotterdam)
Context: Relevantie Data Science Bijlage 05 - Opzet Data Science for logistici (Hogeschool
Rotterdam)
Context: Relevantie Data Science Door de voortgaande informatisering in de maatschappij is er een sterke groei
in de beschikbaarheid van data. Denk hierbij aan data die beschikbaar komt
dankzij systemen zoals ERP, TMS, WMS, IoT en Bonuskaarten. Die toename in het
beschikbaar zijn van data biedt allerlei kansen om middels (statistische)
analyses tot betere informatie te komen en daarmee tot betere adviezen en
besluiten. De beschikbaarheid van die grote hoeveelheden data maakt het
tevens mogelijk om AI (Artificial Intelligence) te verwezenlijken. De groei van de hoeveelheid data is dusdanig dat het bewerken met
traditionele systemen zoals databasemanagementsystemen en spreadsheets
niet goed meer mogelijk is. We spreken dan van Big Data waarbij het gaat om de
grootte van de hoeveelheid data, de snelheid waarmee nieuwe data
binnenkomt en de diversiteit van die data. Middels datawarehousing kan er
voor gezorgd worden dat de gewenste data op de juiste tijd in de juiste
hoeveelheid beschikbaar is. Het gericht zoeken naar verbanden in die Big Data
wordt data mining genoemd. Voor deze cursus gaan we ervan uit dat de
stappen van Big data naar datawarehouse naar datasets al gemaakt zijn. We
gaan wel aan de slag met de stap data mining. Dit is ook de activiteit waar je als
Logistics Engineer het meest mee in aanraking zal komen in het beroepenveld. De volgende onderwerpen komen in deze cursus aan de orde: • De verschillende begrippen welke vallen onder data science
• ERD, Databases
• KPI-Dashboard • Data mining process (aan de hand van CRISP-DM Business Understanding,
Data Understanding, Data Preparation, Modelling, Evaluation, Deployment) • Beschrijvende statistiek (Meetniveaus, populatie, steekproef, frequentie,
variabelen, diagrammen en histogrammen, gemiddelde, mediaan, modus,
variantie, meetkundig gemiddelde) • Beschrijvende statistiek (Meetniveaus, populatie, steekproef, frequentie,
variabelen, diagrammen en histogrammen, gemiddelde, mediaan, modus,
variantie, meetkundig gemiddelde) • Trends, voortschrijdend gemiddelde, correlatie, regressie, tijdreeksen
• Discrete en continue kansvariabelen, zweetkansen
• Kansverdelingen
• Steekproeftrekking, vaststellen aantal te verrichten metingen,
betrouwbaarheidsinterval
• Validatie, Toetsing van hypothesen. • Process mining
• AI
•
Data science en ethiek • Steekproeftrekking, vaststellen aantal te verrichten metingen, Context: de opdrachten 3. Toepassen van process mining op een gegeven dataset middels DISCO
4. Het doorlopen van de data mining cyclus (CRISP-DM) op een dataset
middels Power BI en middels KNIME
5. Toepassen van AI in een eenvoudige logistieke situatie
6. Het verzorgen van een of meerdere presentaties. Bijlage 05 - Opzet Data Science for logistici (Hogeschool
Rotterdam)
Context: Relevantie Data Science Op verzoek van de docent
tijdens de bijeenkomsten uitwerkingen laten zien en toelichting geven aan
de groep medestudenten en beantwoorden van vragen
7. Het geven van feedback op de uitwerkingen van studenten en het stellen
van vragen
8. Het mondeling toelichten van het gemaakte portfolio aan de docent
De activiteiten krijgen vorm door middel van individuele opdrachten en
samenvoegen daarvan in een portfolio. Koppeling met skills (zoals beschreven in paper Logistiek + NR13, tabel 5)
De relatie van deze module met de kerncompetenties van Leng staan in
onderstaand schema. Tabel 2: Leerdoelen en competenties
Leerdoel
Draagt bij aan de
ontwikkeling van de
kerncompetentie
Door middel van 3. Toepassen van process mining op een gegeven dataset middels DISCO 4. Het doorlopen van de data mining cyclus (CRISP-DM) op een dataset
middels Power BI en middels KNIME www.sharehouselab.nl www.sharehouselab.nl www.sharehouselab.nl De student kan:
•
op gestructureerde wijze
een KPI-Dashboard
opstellen middels Power
BI
•
op verantwoorde wijze de
CRISP-DM cyclus
toepassen op een
logistiek data mining
vraagstuk
•
op verantwoorde wijze
een steekproef vaststellen
en de uitkomsten
interpreteren
•
op verantwoorde wijze
process mining toepassen
op een logistiek vraagstuk
•
kan AI
toepassingsmogelijkheden
binnen een logistieke
context ethisch
verantwoord vaststellen
en een eenvoudige AI
toepassing gebruiken en
aanpassen
De student begrijpt de
belangrijkste begrippen die
horen bij data science
A2: Onderzoekt met behulp
van verantwoord gekozen
methoden en technieken
een
(economisch/technisch)
logistiek vraagstuk ter
verbetering/vernieuwing
van het logistiekeproces,
product en/of dienst. Boks algemeen landelijk:
•
Data-analyse
•
Statistische verdelingen
•
Forecasting
•
Kansverdelingen
•
Tijdanalyse
•
Betrouwbaarheidsintervallen
en steekproefgroottes
•
Modelleren
•
Dataverzameling + analyse
•
Betrouwbaarheid en
validiteit
Begrippen:
•
CRISP-DM
•
Power BI, KNIME, DISCO
•
meetniveaus
•
populatie, steekproef
•
frequentie
•
variabelen,
•
diagrammen&histogrammen
•
gemiddelde, mediaan,
modus, variantie,
meetkundig gemiddelde
•
trend, (on)afhankelijkheid,
verdelingsfunctie,
verwachtingswaarde en
variantie, discrete en
continue kansvariabelen
•
lineaire regressie
•
correlatie
•
kansen, kansverdelingen
•
voortschrijdend gemiddelde
•
tijdreeksen
•
validatie
•
toetsing van hypothesen. •
AI
•
Ethiek •
op gestructureerde wijze
een KPI-Dashboard
opstellen middels Power
BI •
op verantwoorde wijze de
CRISP-DM cyclus
toepassen op een
logistiek data mining
vraagstuk www.sharehouselab.nl www.sharehouselab.nl Bijlage 06 – Praktijkcasus STC Het STC heeft naar aanleiding van het onderzoek naar de logistieke skills in een veranderende
warehousecontext het product Vakoverstijgende Casus ontwikkeld, welke gegeven wordt aan
de 2e jaar studenten van de opleiding Logistiek Supervisor mbo niveau 4. De casus raakt de
skills op de gebieden kennis, vaardigheden en gedrag. Met de casus werken de studenten in groepen van 4 samen en worden alle aspecten van de
vakken die door de studenten zijn gevolgd ‘geraakt’. Op die manier zien zij de samenhang en het
belang van de vakken in de context van het beroep. De casus is geschreven met bedrijven
vanuit een diverse logistieke achtergrond. Met de kennis van de stageperiode (1e twee periodes
van het jaar) en de opdracht om kennis te nemen van de bedrijven draagt de casus bij aan de
beeldvorming van ketenlogistiek. Door de opzet, het samenwerken, het communiceren, onderzoek, overleggen (en vastleggen
daarvan) is er ook een groot aandeel soft skills training ondergebracht in deze casus. Een
onderwerp wat uit STC onderzoek al naar voren is gekomen om, vanuit input van het
bedrijfsleven, meer aandacht aan te besteden. Onderstaand staat de opbouw en de leerdoelen aangegeven van de casus: Tijdens jouw leerjaren aan het STC heb je verschillende vakken gevolgd. Waar het op het STC in eerste instantie
individuele vakken waren is het in de praktijk allemaal aan elkaar gekoppeld. De vakoverstijgende casus module
laat jullie zien hoe het er in de praktijk aan toegaat, waarbij je verschillende skills gaat combineren om tot een
goed eindresultaat te komen. Denk hierbij aan: samenwerken relatiebeheer werkoverleg communiceren in een andere taal etc g
Denk hierbij aan: samenwerken, relatiebeheer, werkoverleg, communiceren in een andere taal, etc. WIJZE VAN EXAMINERING EN/OF AFSLUITNG Je wordt tijdens deze module beoordeeld op je aanwezigheid, het samenwerken, en het maken van de diverse
onderdelen. LEERDOELEN EN SUCCESCRITERIA UITLEG Bij ieder leerdoel is beschreven wanneer je het leerdoel succesvol beheerst. Dat is volgens de onderstaande
schaal beschreven. Het begint bij een enkel idee, naar meerdere ideeën, naar de relatie tussen ideeën en de
transfer naar nieuwe ideeën. Bij ieder leerdoel is beschreven wanneer je het leerdoel succesvol beheerst. Dat is volgens de onderstaande
schaal beschreven. Het begint bij een enkel idee, naar meerdere ideeën, naar de relatie tussen ideeën en de
transfer naar nieuwe ideeën. www.sharehouselab.nl Bijlage 07 – Casus Deltion E-logistics www.sharehouselab.nl Bijlage 07 – Casus Deltion E-logistics Context: De logistieke sector groeit hard en is volop in beweging. Op alle
opleidingsniveaus (MBO, HBO, WO) veranderen banen in de logistiek vanwege
technologische ontwikkelingen en digitaliseringsvraagstukken. Dat vraagt om
een vernieuwde visie op ontwikkelen van human capital waarin leren, werken en
innoveren nauwer met elkaar verbonden zijn. Vanuit deze achtergrond is deze onderwijscasus ontwikkeld. In deze casus gaan de MBO & HBO - studenten samenwerken in een fysieke
logistieke omgeving, waarbij het van belang is dat MBO studenten inzicht
krijgen in wat HBO studenten doen en hoe ze werken (in het kader van
eventuele doorstroom naar het hoger onderwijs, i.e. AD of HBO onderwijs). Anderzijds is het van belang dat HBO studenten inzicht krijgen en vaardiger
worden in hoe zij de MBO studenten kunnen betrekken in hun verbeterproject. Leerdoelen/uitkomsten: Deze onderwijscasus kent meerdere leerdoelen, waarbij de essentie bestaat uit
het samenwerken en leren tussen MBO en HBO studenten. Leerdoelen waaraan
gewerkt worden zijn; Deze onderwijscasus kent meerdere leerdoelen, waarbij de essentie bestaat uit
het samenwerken en leren tussen MBO en HBO studenten. Leerdoelen waaraan
gewerkt worden zijn; -
Begrip krijgen voor de verschillen in competentieniveau vanuit zowel het MBO als
HBO perspectief. -
De verschillende rollen kennen en herkennen vanuit een (LEAN-) verbeterproject -
De verschillende rollen kennen en herkennen vanuit een (LEAN-) verbeterproject. Leerdoelen MBO-studenten Vanuit de MBO-studenten kunnen de volgende leerdoelen worden
gedefinieerd. • Het aanleren van diverse vaardigheden in de inslag-, opslag- en uitslag
werkzaamheden. • Leren wat het betekent om onderdeel uit te maken van een (LEAN-) verbeterproject. • Kunnen omgaan met druk. • Kritisch worden op het proces waar je zelf onderdeel van uitmaakt. • Basis vaardigheden aanleren WMS-systeem. • Aanleren om te kunnen gaan met veranderende werkomstandigheden. (toekomstgericht) Toetsing g
In de huidige vorm vindt de toetsing plaats door middel van een adviesportfolio
en een presentatie waarin de HBO-studenten een advies opleveren waarin de
verschillende leerdoelen zijn verwerkt en waarbij nadrukkelijk zichtbaar wordt
gemaakt op welk manier de medewerkers (de MBO studenten dus) worden
betrokken. Leerdoelen HBO-studenten Vanuit de HBO-studenten kunnen de volgende leerdoelen worden gedefinieerd. De HBO-studenten leren; Vanuit de HBO-studenten kunnen de volgende leerdoelen worden gedefinieerd. De HBO-studenten leren; -
wat het betekent een (LEAN-) verbeterproject zelf vorm te geven en in
groepsverband te faciliteren. -
inzien welke aanpak werkt om de MBO-student in een ‘kritische’ houding te krijgen
met betrekking tot het proces -
een goede probleemstelling te definieren -
een goede probleemstelling te definieren -
een dataverzamelingsplan op te stellen en daarbij zelf ook goede metingen te
ontwerpen -
een procesflow visueel weer te geven welke begrijpelijk is voor de MBO-studente -
oorzaak-gevolg relaties begrijpelijk weer te geven en gezamenlijk met MBO-
studenten af te kunnen stemmen -
middels brainstorm technieken gezamenlijk met de MBO-studenten verbeter ideeën
opstellen -
een afweging maken welke verbeter idee het beste is voor de organisatie, waarbij
rekening wordt gehouden met people, planet, profit. -
een business case voor één van de verbeter ideeën uitwerken -
een voorstel doen hoe het verbetervoorstel geborgd kan worden www.sharehouselab.nl Bijlage 08 – Lean Orange Belt Bijlage 08 – Lean Orange Belt Heb je je wel eens afgevraagd welke processen je achter de schermen allemaal
in gang zet als: • Je schoenen bestelt via een webwinkel? • Je een abonnement afsluit bij een aanbieder van je favoriete series en films • Een student een herkansing aanvraagt via de examencommissie? Om een dienst te verlenen of een product te maken zijn processen nodig. Organisaties bestaan omdat ze waarde toevoegen voor de klant. Vind jij het
leuk om samen na te denken hoe je processen slimmer kunt organiseren? In dat
geval is Lean Management interessant voor jou. • Een student een herkansing aanvraagt via de examencommissie? Om een dienst te verlenen of een product te maken zijn processen nodig. Organisaties bestaan omdat ze waarde toevoegen voor de klant. Vind jij het
leuk om samen na te denken hoe je processen slimmer kunt organiseren? In dat
geval is Lean Management interessant voor jou. De Orange Belt in Lean geeft je een ‘kijkje in de keuken’ van Lean. De MBO studenten krijgen veelal informatie over de theorie van uitvoerende
werkzaamheden, echter wordt hen nog weinig handvatten gegeven om
gestructureerd aan de slag te gaan met procesverbeteringen. Op het Logistieke
MBO niveau zijn de studenten goed in staat om de goederenstromen te
herkennen en soms zelf te verbeteren. Maar hoe zorg jij er nu voor dat je ook in
staat bent om continu (kleine stapjes) te verbeteren. Hoe moet je dan naar
processen kijken, welke tools en technieken kunnen je daar dan bij helpen en
hoe zorg je er ook voor dat je de organisatie hierin goed kunt meenemen en
uiteindelijk ervoor zorgt dat de verbeteringen ook continu op de voorgestelde
manier worden uitgevoerd? Van de Lean-filosofie is het continu verbeteren in kleine stappen een manier
van denken waarmee tevens sociale innovatie vanaf de werkvloer meer en meer
wordt gestimuleerd. Immers, elke dag een kleine verbetering vanaf de werkvloer
levert sneller resultaat dan één omvangrijk project. www.sharehouselab.nl www.sharehouselab.nl Leerinhoud: We starten met een introductie van Lean. Je maakt kennis met de achtergrond
en uitgangspunten van het Lean gedachtengoed. Daarin staat het toevoegen
van waarde voor een klant centraal. Je leert vervolgens hoe je samen met collega’s stapsgewijs eenvoudige
processen kunt visualiseren, analyseren, verbeteren en borgen. Dat doe je aan
de hand van een doorlopende casus. Daarnaast pas je deze inzichten toe in je eigen werk. Je gaat op Safari om
verspillingen in je eigen werkomgeving te herkennen. Je maakt kennis met de
A3 methode waarmee je een kleine procesverbetering gaat uitwerken. Er zijn 3 bijeenkomsten van 4 uur, met de volgende inhoud Er zijn 3 bijeenkomsten van 4 uur, met de volgende inhoud: • Theorie deel I, casus en introductie opdracht Lean Safari • Theorie deel II, casus, game en introductie opdracht A3 • Werkbijeenkomst A3 procesverbetering Na deze bijeenkomsten wordt een show & tell bijeenkomst gepland. Daarin
presenteert elke deelnemer zijn of haar A3 poster over een eigen
procesverbetering in het bijzijn van medecursisten. Leidinggevenden zijn ook
welkom. Leerdoelen/uitkomsten: Na deze module ben je in staat om zelf gestructureerd kleine verbeteringen te
kunnen initiëren. En kun je deelnemen in een groter project, waarbij
verschillende Lean-elementen worden toegepast. Deze module geeft handvatten om deel te nemen aan een procesverbetering. Ook leer je om zelf ‘kritisch’ en gestructureerd te kijken naar je ‘eigen’ (werk-
)processen. www.sharehouselab.nl Toetsing g
Deze training wordt afgerond met het inleveren van een ‘ingevulde A3-poster’
(Format Orange Belt). Hiermee toon je aan dat je actief met Lean bezig bent
geweest en dat je je hebt ontwikkeld in het vakgebied van proces verbeteren. www.sharehouselab.nl www.sharehouselab.nl Bijlage 09 – De lerende organisatie Bijlage
Context Context Technologische innovatie heeft een toevlucht genomen. Termen als
automatisering en robotisering zijn inmiddels gemeengoed geworden in de
logistieke sector. Maar inmiddels gaan we alweer een stap verder. Een
ontwikkeling die niet wordt gekenmerkt door de opkomst van een enkele
nieuwe technologie, maar door een samengaan van diverse technologieën en
disciplines, zoals kunstmatige intelligentie, robotica, biotechnologie en nieuwe
maakmethoden (zoals 3D-printen). Samen zorgen ze voor ongekende nieuwe
mogelijkheden en veranderen ze het proces van ontwerpen, produceren en
consumeren ingrijpend. Toch is het invoeren, adopteren of integreren van deze nieuwe technologieën
en disciplines voor veel organisaties een uitdaging. Niet alleen hebben
dergelijke veranderingen impact op de supply chain en de primaire processen
binnen een organisatie, maar ook worden andere skills en vaardigheden van
medewerkers verwacht. Sociale innovatie is als belangrijkste enabler van
technologische innovatie ondenkbaar geworden. Je kunt nog zo hard
technologische vernieuwing inzetten, als medewerkers deze niet zelf willen
inzetten en ze willen adopteren dan gaat het nooit lukken. Dit vraagt om een lerende organisatie, een organisatie die niet alleen
technologisch innovatief is, maar ook sociaal innovatief. Een organisatie waar
men flexibel is om verandering te adopteren, eigen te maken en te realiseren. En waar op alle niveaus wordt geleerd en kennis en vaardigheden worden
ontwikkeld en gedeeld. Het is een model waar veel organisaties warm voor
worden, maar ze hebben veelal geen idee hoe dat moet. In deze module wordt vanuit verschillende invalshoeken (het individu, het team,
de organisatie/keten) de lerende organisatie belicht met als doel in staat te zijn
een lerende cultuur in organisaties te creëren, waarin medewerkers zelf aan de
slag gaan om proces- en technologische innovaties te realiseren. Leerdoelen/uitkomsten: www.sharehouselab.nl Leerinhoud: 1. Het verkrijgen van kennis op gebied van diverse thema’s omtrent de lerende
organisatie. 2. Het toepassen van kennis ten aanzien van een lerende organisatie 3. Het analyseren van aspecten van een lerende organisatie m.b.v. kennis bij een
organisatie 4. Het formuleren van een interventiestrategie gericht op het vergroten van het lere
binnen organisaties 5. Samenwerken en vergroten van adviesvaardigheden en sociale vaardigheden De opdracht Studenten verdiepen zich in het onderwerp en schrijven een visie ten aanzien
van het onderwerp. Op basis van de visie wordt er een GAP-analyse uitgevoerd
bij een organisatie. Aansluitend formuleren de studenten een
interventieprogramma waarmee het leren kan worden gestimuleerd. De activiteiten krijgen vorm door middel van groepsopdrachten en
samenvoegen daarvan in een portfolio. De volgende onderwerpen komen aan de orde: De volgende onderwerpen komen aan de orde: • Inleiding: Lerende organisatie, waaronder sociale innovatie • Medewerker van de toekomst, skills & vaardigheden, drijfveren en motivatie,
werkgeluk • Persoonlijk leiderschap, learning agility, duurzame inzetbaarheid • Werkplekleren, training within industry, leercultuur, zelforganisatie en
zelfsturende teams • Transformationeel leiderschap, coachend leiderschap, dienend leiderschap
en gedeeld leiderschap • Co-creatie, ketenregie • Interventies om leren te stimuleren • Interventies om leren te stimuleren De leerdoelen geven aan wat studenten aan het eind van de leerlijn moeten
beheersen De leerdoelen geven aan wat studenten aan het eind van de leerlijn moeten
beheersen De student weet uit welke componenten een lerende organisatie bestaat en op
welke manier leren in een organisatie kan worden gestimuleerd en gefaciliteerd
en kan hierbij een visie/gewenste situatie formuleren waarin een lerende
organisatie wordt beschreven. De student is in staat een GAP-analyse te maken
binnen een organisatie waarin wordt beschouwd welke componenten aanwezig www.sharehouselab.nl zijn in de huidige situatie van de organisatie en kan suggesties geven voor
interventies waardoor het leren wordt gestimuleerd. zijn in de huidige situatie van de organisatie en kan suggesties geven voor
interventies waardoor het leren wordt gestimuleerd. Leerinhoud: Leeruitkomsten M1 1. De student bereid een kennismakingsgesprek met een organisatie voor door zich te
verdiepen in deze organisatie. De student voert het kennismakingsgesprek waarin hij/zij
gebruik maakt van luister- en basisgespreksvaardigheden en erkende gesprekstechnieken. De
student speelt in op non-verbale communicatie en herkent de communicatiestijl van de
gesprekspartners en past de eigen stijl erop aan. 2. De student presenteert en ontvangt hierover feedback. De student maakt hierbij gebruik van
basispresentatievaardigheden. Hij/Zij heeft een open houding ten aanzien van de ontvangen
feedback en reflecteert hier kritisch op. De student geeft op een passende wijze feedback op
de presentatievaardigheden van een of meerdere peers. 2. De student presenteert en ontvangt hierover feedback. De student maakt hierbij gebruik van
basispresentatievaardigheden. Hij/Zij heeft een open houding ten aanzien van de ontvangen
feedback en reflecteert hier kritisch op. De student geeft op een passende wijze feedback op
de presentatievaardigheden van een of meerdere peers. 1. De student kan kwaliteiten en valkuilen van zichzelf en anderen herkennen en analyseren,
met betrekking tot teamvorming tijdens de samenwerking. Hierbij wordt de gekozen
projectmanagementmethode gehanteerd. De studenten maken gebruik van ieders kracht
wat leidt tot acceptatie van de verschillen tussen alle groepsleden. 2. De student schrijft een stappenplan met betrekking tot het organiseren van toekomstige
projecten. In het stappenplan worden teamrollen en teamvorming behandelt en maken
gebruik van een projectmanagementmethode wat leidt tot inzicht in het organiseren van een
samenwerking. 3. De student regisseert zijn of haar eigen professionele ontwikkeling, met betrekking tot het
kernonderwerp samenwerken, door het verzamelen van bewijslast en te reflecteren aan de
hand van een zelfgekozen reflectiemodel, waaruit de student twee SMART leerdoelen
formuleert ten behoeve van toekomstige ontwikkeling. www.sharehouselab.nl ijlage 10 - Professionele Ontwikkeling leerjaar 1 (Semester 1 & 2 Context Als de logistieke procesverbeteraar van de toekomst is het van essentieel belang dat
studenten theoretische kennis in de praktijk kunnen toepassen. Ter ondersteuning
hieraan wordt er gedurende de hele opleiding aandacht besteed aan de professionele
ontwikkeling van de studenten. Hierin staan 3 kernonderwerpen centraal: o Samenwerken o Adviseren o Omgaan met weerstand & draagvlak creëren
In het eerste leerjaar van de opleiding gaan studenten bezig met het kernonderwerp
Samenwerken. Tijdens semester 1 staan communicatieve vaardigheden centraal. Deze
hebben studenten nodig voor semester 2. In semester 2 staat samenwerken centraal. www.sharehouselab.nl www.sharehouselab.nl De volgende leerinhoud wordt aangeboden tijdens PO M1: o Communicatiemodellen
o (Non)verbale communicatie
o Miscommunicatie
o Communicatiestijlen
o Communicatieniveaus o Communicatiemodellen www.sharehouselab.nl o Gedrag tijdens communicatie o Gedrag tijdens communicatie o Feedback geven en ontvangen Context: de opdracht M1 Tijdens PO M1 bereiden we de studenten voor op het assessment die zij aan het einde
van de periode afleggen. Aan het eind van periode 1 betreft dit gesprekstechnieken. In
periode 2 betreft dit presenteren in combinatie met feedback geven en ontvangen. De volgende leerinhoud wordt aangeboden tijdens PO M2: De volgende leerinhoud wordt aangeboden tijdens PO M2: o Ieders kracht gebruiken
o Aanpassingsvermogen
o Besluitvorming
o Overtuigen
o Teamrollen
o Gedragsprofielen
o Fasen in teamvorming
o Interculturele verschillen o Ieders kracht gebruiken Context: de opdracht M2 Studenten krijgen verwerkingsopdrachten om bewustwording te creëren van hun eigen
rol binnen een team. Daarnaast kennen zij hun kwaliteiten en valkuilen en kunnen
hierdoor een Persoonlijk Profiel maken. Dit Persoonlijk Profiel gebruiken studenten
tijdens de samenwerkingsopdracht die zij in een looptijd van 4 weken gaan uitvoeren. Ondertussen krijgen studenten handvatten mee over teamvorming, doelen formuleren,
overtuigen, besluitvorming en aanpassingsvermogen. Deze aspecten passen zij direct
toe in de samenwerkingsopdracht. Vervolgens gaan zij in periode 4 de praktijk in en
verzamelen bewijslast waarin zij laten zien aan de vaardigheid te hebben gewerkt en
reflecteren op hun ontwikkeling. Dit dient ter voorbereiding op BUCA in leerjaar 2. Deze
opdrachten worden samengevoegd in het portfolio: Evaluatie & Ontwikkeling. www.sharehouselab.nl www.sharehouselab.nl
|
https://openalex.org/W4224002938
|
https://jurnal.uns.ac.id/mekanika/article/download/51479/35415
|
English
| null |
Nanofiber Semiconductor Experiment with Distance Variation Electrospinning Method to Improve DSSC Performance
|
Mekanika
| 2,022
|
cc-by
| 3,414
|
Mekanika: Majalah Ilmiah Mekanika Mekanika: Majalah Ilmiah Mekanika Volume 19 (1) 2020
Volume 21 (1) 2022 Volume 19 (1) 2020
Volume 21 (1) 2022 Page 1 - 6
Page 13 - 19 Page 1 - 6
Page 13 - 19 Nanofiber Semiconductor Experiment with Distance Variation
Electrospinning Method to Improve the Efficiency of the DSSC
Denny Widhiyanuriyawan1, Zainal Arifin2*, Syamsul Hadi2, Muhammad Rijalul Fadli2
1 Department of Mechanical Engineering, Universitas Brawijaya, Malang, Indonesia
2 Department of Mechanical Engineering, Universitas Sebelas Maret, Surakarta, Indonesia
*Corresponding Author’s email address: zainal_arifin@staff.uns.ac.id Keywords:
DSSC
Electrospinning
Rotating collector
Distance Abstract
Dye-Sensitized Solar Cell (DSSC) is a solar cell that uses dyes to transfer sunlight
to electrical energy. DSSC construction uses a layered system (sandwich) that
consists of a working electrode and an opposing electrode, both of which are placed
on conducting glass and electrolytes to allow electron cycling. DSSC has a high
price, so increasing efficiency is needed to use these solar cells more widely. This
research aims to determine the effect of the distance between the tip and the rotating
collector to increase the efficiency of the Dye-Sensitized Solar Cell (DSSC) and
examine its impact on the morphology of the ZnO nanofiber. This experiment is
carried out by varying the distance between the tip to the rotating collector, 4 cm, 6
cm, and 8 cm. This research indicates that at a distance of 8 cm, it produces a small,
uniform, and stable ZnO nanofiber structure with Voc, Jsc, FF, and DSSC efficiency
values of 0.559 V, 9.809 mA/cm2, 43.3%, and 2.3%. In addition, at a distance of 8
cm, it also produces the highest DSSC electrical efficiency from the other distances
due to the absorbance of the dye and high electron excitation. Revised 27 May 2021; received in revised version 20 February 2022; Accepted 21 March 2022
Available Online 31 March 2022 Widhiyanuriyawan et al. Gratzel's research shows that DSSC has a relatively low price compared to the previous generation,
simple fabrication, and environmentally friendly materials [4]. DSSC consists of five layers: Fluorine-
doped Tin Oxide (FTO) glass, semiconductor, dye, electrolyte, and counter electrode [5]. The efficiency of
the DSSC depends on the semiconductor that must have high electron mobility and a surface area to absorb
the dye. Currently, DSSC has an efficiency of 15% [6]. Semiconductor materials that are often used in
DSSC are TiO2 and ZnO. Both have their advantages and disadvantages, including the advantages of ZnO,
namely having a wide band gap of 3.37 eV [7] and the high electron mobility of TiO2 [8]. At the same time,
the drawback is the level of chemical stability which is relatively low. TiO2 has advantages, namely high
color adsorption ability and high chemical stability but the disadvantage of TiO2 is that it has a low band
gap of 3.2 eV [7]. To get optimal performance, the Double Layer method is used. This method is used to
form a DSSC anode using several semiconductor layers to form a DSSC anode order to have good
performance. The electrospinning method with a rotating collector makes nanofiber fibers with an easy
working principle and can maintain the uniformity of the nanofiber produced. In principle, electrospinning
utilizes the electrostatic force that arises from charged particles due to a high voltage between the tip of the
tip and the rotating collector [9]. Photoanode of DSSC made of nanomaterial with different morphologies are nanofibers, nanowires,
nanorods, nanobelts, nanotubes, nano spiral, nano rings [10], and nanoparticles [11]. Nanofiber has the
advantage that nanofiber morphology can provide direct electron flow from photogeneration to the
conductive substrate. The branching structure makes the morphology of the nanofiber has a high surface
area. Nanofiber structure that can stick to the substrate and the dye will directly facilitate the mobility of
electrons that have been excited. The direct deposition method is a coating technique on nanomaterials with
an electrospinning process in which the precursor solution is sprayed directly onto the FTO glass. The
benefit of this method is that the ZnO coating process on the DSSC assembly will be shorter and reduce
nanofiber damage. 1 Introduction Energy is the primary source of life in the world. The use of energy in society still depends on non-
renewable fossil fuels such as petroleum, coal, and natural gas. This fossil fuel is not only limited in use
but also can cause environmental pollution. To overcome this problem, another alternative fuel is needed. Therefore, researchers focused on clean and renewable energy sources, namely solar energy. Solar energy
is a source of energy that produces heat and radiation. Solar cells are a tool that can convert solar radiation
into electrical energy through a photovoltaic process [1,2]. Solar cells have been developed from generation to generation with different characters and different
components, namely silicon-based solar cells, thin film-based solar cells (thin film), and DSSC. Silicon-
based solar cells are made of silicon and have very high efficiency. The disadvantage of this solar cell is
that it is expensive to produce. Thin film-based solar cells are a development of silicon solar cells, which
have lower efficiency than silicon solar cells but are more economical. The price is lower than silicon solar
cells [3]. Michael Gratzel first discovered the Dye-Sensitized Solar Cell (DSSC) solar cell in 1991. https://dx.doi.org/10.20961/mekanika.v21i1.51479 https://dx.doi.org/10.20961/mekanika.v21i1.51479 2579-3144
© 2022 Mekanika: Majalah Ilmiah Mekanika. All right reserved 2579-3144
© 2022 Mekanika: Majalah Ilmiah Mekanika. All right reserved 13 Volume 21 (1) 2022 Widhiyanuriyawan et al. Meanwhile, the attachment process to nanomaterials such as doctor blade, slip casting,
and spin coating uses a similar technique by coating the nanomaterial from a paste form into a thin layer
on FTO glass. The process before coating the nanomaterial paste is by making a solution first. Then the
solution is converted into a nanomaterial that has been sintered [12]. This method still has some
disadvantages, such as a more extended procedure and can damage the nanofibers. In this study, the DSSC photoanode manufacturing process will be examined using distance variations
in the electrospinning process with the direct deposition method on a rotating collector and a method of
coating nanomaterials using ZnO material with an electrospinning process where the results of spraying
ZnO solution will be captured directly by the FTO which has been coated with TiO2 nanoparticles-
moreover, glued to the rotating collector. Therefore, the direct deposition method using electrospinning
with a rotating collector will shorten the DSSC fabrication process and hopefully reduce the damage to the
nanofiber produced and produce a good performance. Volume 21 (1) 2022 Volume 21 (1) 2022 Volume 21 (1) 2022 Widhiyanuriyawan et al. remove the formed foam [13,14] so that a Zn(Ac)2/PVA solution is produced, which is used to produce
ZnO nanofiber fibres using an electrospinning machine with a rotating collector as shown in Figure 1. Figure 1. Schematic illustration of electrospinning with rotating collector [15] Figure 1. Schematic illustration of electrospinning with rotating collector [15] The solution that has been synthesized is then put into 1 ml of the electrospinning solution pump
injection. The central part of the electrospinning device consists of a high-voltage Direct Current (DC)
source, a spinneret tube, and a small diameter needle with an inner diameter of 200 µm, which is connected
to the positive 15 kV high voltage terminal to the plate on the rotating collector made of metal to which
FTO glass has been deposited with TiO2. Nanoparticles and connected to the negative terminal using the
direct deposition method (Figure 1). The flow rate of the solution through the needle is 4 µL / minute
because it can produce more uniform nanofiber fibers. The tip distance to the rotating collector uses a
variation of 4, 6, 8 cm. the rotating speed at the collector is 45 rpm. The solution then attaches itself to the
surface of the collector plate in the form of pulling strands called green fibers. The green fibers were then
centered at a temperature of about 500 ℃ with one hour of holding to remove organic matter and form ZnO
crystals [16]. The result of nanofiber deposition on FTO coated with TiO2 nanoparticles can be used as a
double layer photoanode. 2.1 Preparation of photoanode This research uses semiconductor types of ZnO nanofiber and TiO2 nanoparticles. To obtain the
nanofiber morphology, an electrospinning process was carried out with a rotating collector so that
semiconductors need to be added to precursor solutions which are polymers with good electrical
conductivity. Preparation of precursors using 10 grams of Polyvinyl Alcohol (PVA) (Merck, MW = 72,000)
mixed with 10 ml of distilled water (H2O). To produce a homogenized solution, the stirring process was
carried out for four hours at a solution temperature of 70 ℃. The Zn(Ac)2 solution was synthesized using 2
grams of zinc acetate dihydrate ((CH3COO)2Zn.2H2O, Merck) and 8 ml of H2O, after which the
homogeneity process was carried out by stirring for 1 hour, keeping the solution at 70 ℃. The
homogenization process between the PVA and Zn(Ac)2 solution was carried out in a 4:1 wt% at 70 ℃ for
eight hours. Furthermore, the Zn(Ac)2/PVA solution was left to stand at room temperature for 24 hours to 14 2.2 DSSC fabrication and testing process After the sintering process is carried out, the FTO is left to cool then soaked in sanitizer-N719 dye
([RuL2 (NCS) 2]: 2TBA (L = 2,2'-bipyridyl-4,4'-dicarboxylic acid; TBA = tetra- n-butylammonium)) from
Dyesol for one day. The DSSC fabrication process combines the FTO with a counter electrode, including
semiconductors and dye. Then, the electrolyte solution is placed between the FTO and the counter electrode
using the injection method. The electrolyte used in this study was Iodide (I3-) EL-HPE from Dyesol. In
general, the performance of the DSSC can be characterized by several electrical parameters, including the
Photocurrent-Voltage curve (JV curve), Open-Circuit Photovoltage (Voc), and Short-Circuit Photocurrent
Density (Jsc), Fill Factor (FF), and Efficiency (η). The measurement is carried out by shining the DSSC at
the measured intensity (generally 100 mW/cm2) and given a variation of the electric load from zero to
infinity. Then the voltage and current are measured every time [18]. The results of the measurements are in
the form of voltage and current values, which are plotted in a graph as in Figure 2. 15 Widhiyanuriyawan et al. Volume 21 (1) 2022
Figure 2. Current – voltage curve (I–V curve) on DSSC [17] Volume 21 (1) 2022 Widhiyanuriyawan et al. Figure 2. Current – voltage curve (I–V curve) on DSSC [17] Figure 2. Current – voltage curve (I–V curve) on DSSC [17] From the voltage-current curve (Figure 2), it can be seen that the Fill Factor (FF) value, that the fill
factor is a parameter that determines the efficiency of the DSSC. Graphically, the fill factor is the ratio
between the maximum power (PMPP) to the multiplication of short-circuit current (Isc) and open-circuit
voltage (Voc). To find out the fill factor (FF) value, you can use Equation 1 [17]. 𝐹𝐹= 𝑉𝑀𝑃𝑃 𝑥 𝐼𝑀𝑃𝑃
𝐼𝑆𝐶 𝑥 𝑉𝑂𝐶
(1) 𝐹𝐹= 𝑉𝑀𝑃𝑃 𝑥 𝐼𝑀𝑃𝑃
𝐼𝑆𝐶 𝑥 𝑉𝑂𝐶 (1) The efficiency of the DSSC (Equation 2) solar cell is the ratio between the maximum power (PMPP) to the
power emitted by light in the active region of the solar cell (Plight). Which was (Plight) is the multiplication
of sunlight intensity (Irad) to the active area of the solar cell (A) [18]. 𝜂= 𝑃𝑀𝑃𝑃
𝑃𝑙𝑖𝑔ℎ𝑡
= 𝑃𝑀𝑃𝑃
𝐼 𝑥 𝐴= 𝐼𝑆𝐶 𝑥 𝑉𝑂𝐶 𝑥 𝐹𝐹
𝐼 𝑥 𝐴
(2) (2) 3.1 DSSC nanofiber morphology test 3.1 DSSC nanofiber morphology test Figure 3 shows the results of the Scanning Electron Microscopy (SEM) ZnO nanofiber test by using
the shorter tip distance to the rotating collector, which will result in an irregular morphology of the
nanofiber, has a large diameter and has branches like dendrites on nerves. This situation occurs due to poor
solvent evaporation, which causes irregular beading and build-up of nanofiber, as shown in Figure 3 (a). The shorter the tip distance to the rotating collector, the less solvent evaporation process, so that the solvent
adhering to the Fluorine-doped Tin Oxide (FTO) glass is less than perfect. This is consistent with previous
research, which states that the distance between the tip and the rotating collector has sufficient evaporation
time for the nanofiber to fully adhere to the FTO glass, resulting in a thinner and uniform nanofiber. Meanwhile, with the decreasing distance between the tip and the rotating collector, the resulting nanofiber
is irregular, the nanofiber becomes branched, and there are lumps in the resulting nanofiber. 16 Volume 21 (1) 2022 Widhiyanuriyawan et al. Figure 3. SEM test results for ZnO nanofiber with variations in tip distance to the rotating collector: (a) 4 cm, (b) 6
cm, and (c) 8 cm with a flow rate of 4 µL/minute
3 2 DSSC
f
(a)
(b)
(c) (a)
(b) (b) (a) (a) (b) (c) (c) Figure 3. SEM test results for ZnO nanofiber with variations in tip distance to the rotating collector: (a) 4 cm, (b) 6
cm, and (c) 8 cm with a flow rate of 4 µL/minute 3.2 DSSC performance The voltage-current curve shows the performance of Dye-Sensitized Solar Cells (DSSC) solar cells
at a tip to collector rotating distance of 4.6 and 8 cm. It is known that the voltage-current curve on the DSSC
is shown in Figure 4. Some of the DSSC performance parameters include Short-Circuit photocurrent
density (Jsc) of 5 mA/cm, Open-Circuit photovoltage (Voc) of 0.658 V, Fill Factor (FF) of 0.29%, and
efficiency of 0.98% for the tip distance to the rotating collector of 8 cm which can produce the highest
current. However, the efficiency values at the tip to collector rotating distances of 4 cm and 6 cm are shown
in Table 1. The results of this study indicate that the Jsc value affects the resulting efficiency value. The
more the Jsc value increases, the DSSC efficiency value will also increase. 17
Figure 4. Current – Voltage curve (I – V curve) on DSSC
0
1
2
3
4
5
6
7
8
9
10
11
0
0.2
0.4
0.6
I (mA)
Voltage (V)
4 cm
6 cm
8 cm I (mA) 17
Figure 4. Current – Voltage curve (I – V curve) on DSSC 17 Volume 21 (1) 2022 Widhiyanuriyawan et al. Table 1. The performance of DSSC
Distance
(cm)
Voc
(V)
Jsc
(mA/cm2)
FF
(%)
η
(%)
4
0.544
7.1
48
1.91
6
0.566
8.1
45.8
2.11
8
0.559
9.8
43.3
2.37 Table 1. The performance of DSSC 4 Conclusions The ZnO nanofiber fabrication process in making photoanodes with nanofiber fibers in DSSC was
carried out using an electrospinning machine with the use of variations in the distance between the tip to
the rotating collector in the electrospinning process using the direct deposition method. The addition of the
distance in the electrospinning process will produce nanofiber fibers with a small and uniform diameter and
size to improve the performance of DSSC because the distribution of electrons becomes more even. Vice
versa, the larger the size and diameter of the resulting nanofiber fibers, the lower the performance of the
DSSC. DSSC performance is also influenced by the value of Jsc. The higher the value of Jsc, the higher the
efficiency of DSSC. In this study, the highest efficiency of the DSSC was 2.37, which was achieved at a
distance of 8 cm. The Voc, Jsc, and FF values were 0.559 V, 9.8 mA/cm2, and 43.3%. References 1. W. Liu, H. Ma, and A. Walsh, “Advance in photonic crystal solar cells,” Renew. Sustain. Energy Rev., vol. 116, article no. 109436, 2019. 1. W. Liu, H. Ma, and A. Walsh, “Advance in photonic crystal solar cells,” Renew. Sustain. Energy Rev., vol. 116, article no. 109436, 2019. 2. S. D. Prasetyo, A. R. Prabowo, and Z. Arifin, “The effect of collector design in increasing PVT performance:
Current state and milestone,” Mater. Today Proc., vol. 63, pp. S1-S9, 2022. 2. S. D. Prasetyo, A. R. Prabowo, and Z. Arifin, “The effect of collector design in increasing PVT performance:
Current state and milestone,” Mater. Today Proc., vol. 63, pp. S1-S9, 2022. 3. Z. Arifin, S. Hadi, H. N. Jati, S. D. Prasetyo, and Suyitno, “Effect of electrospinning distance to fabricate ZnO
nanofiber as photoanode of dye-sensitized solar cells,” AIP Conf. Proc., vol. 2217, article no. 030095, 2020. 4. T. Ponken, S. Kongsankham, S. Panya, W. Choawunklang, and P. Ardchongthong, “Effect of Titanium
Dioxide (TiO2) Light Scattering Layer deposited by Spray Deposition Method at Room Temperature for Dye-
sensitized Solar Cell (DSSC),” Mater. Today Proc., vol. 17, pp. 1249-1258, 2019. 4. T. Ponken, S. Kongsankham, S. Panya, W. Choawunklang, and P. Ardchongthong, “Effect of Titanium
Dioxide (TiO2) Light Scattering Layer deposited by Spray Deposition Method at Room Temperature for Dye-
sensitized Solar Cell (DSSC),” Mater. Today Proc., vol. 17, pp. 1249-1258, 2019. M. K. Nazeeruddin, E. Baranoff, and M. Grätzel, “Dye-sensitized solar cells: A brief overview,” S
ol. 85, no. 6, pp. 1172-1178, 2011. vol. 85, no. 6, pp. 1172-1178, 2011. 6. K. K. Tehare, S. T. Navale, F. J. Stadler, Z. He, and H. Yang, “Enhanced DSSCs performance of TiO2
nanostructure by surface passivation layers,” Mater. Res. Bull., vol. 99, pp. 491-95, 2018. K. K. Tehare, S. T. Navale, F. J. Stadler, Z. He, and H. Yang, “Enhanced DSSCs performan
anostructure by surface passivation layers,” Mater. Res. Bull., vol. 99, pp. 491-95, 2018. 7. B. Boro, B. Gogoi, B. M. Rajbongshi, and A. Ramchiary, “Nano-structured TiO2/ZnO nanocomposite for
dye-sensitized solar cells application: A review,” Renew. Sustain. Energy Rev., vol. 81, pp. 2264-2270, 2018. 8. S. Suhaimi, M. M. Shahimin, Z. A. Alahmed, J. Chyský, and A. H. Reshak, “Materials for enhanced dye-
sensitized solar cell performance: Electrochemical application,” Int. J. Electrochem. Sci., vol. 10, no. 4, pp. 2859-2871, 2015. 9. T. Wahyudi and D. 5 Acknowledgement This work was partially supported by a PDUPT grant from the Ministry of Research, Technology,
and Higher Education, the Republic of Indonesia, with contract number 112/UN27.21/HK/2020 for FY
2020. References Sugiyana, “Pembuatan Serat Nano Menggunakan Metode Electrospinning,” Arena Tekstil,
vol. 26, no. 1, pp. 29-34, 2011. (in Indonesia). 10. S. Yun and S. Lim, “Effect of Al-doping on the structure and optical properties of el
nanofiber films,” J. Colloid Interface Sci., vol. 360, no. 2, pp. 430-439, 2011. d S. Lim, “Effect of Al-doping on the structure and optical properties of electrospun zinc oxid
ilms,” J. Colloid Interface Sci., vol. 360, no. 2, pp. 430-439, 2011. ,
f
pp
11. A. E. R. Mohamed and S. Rohani, “Modified TiO2 nanotube arrays (TNTAs): Progressive strategies towards
visible light responsive photoanode, a review,” Energy Environ. Sci., vol. 4, no. 4, pp. 1065-1086, 2011. 11. A. E. R. Mohamed and S. Rohani, “Modified TiO2 nanotube arrays (TNTAs): Progressive strategies towards
visible light responsive photoanode, a review,” Energy Environ. Sci., vol. 4, no. 4, pp. 1065-1086, 2011. 12. B. D. Li and Y. Xia, “Electrospinning of Nanofibers : Reinventing the Wheel ?” Adv. Mater., vol. 16, no. 14,
pp. 1151-1170, 2004. g
p
p
,
,
gy
,
,
, pp
,
12. B. D. Li and Y. Xia, “Electrospinning of Nanofibers : Reinventing the Wheel ?” Adv. Mater., vol. 16, no. 14,
pp. 1151-1170, 2004. 13. S. Suyitno, A. Purwanto, R. L. L. G. Hidayat, I. Sholahudin, M. Yusuf, S. Huda, and Z. Arifin., “Fabrication
and characterization of zinc oxide-based electrospun nanofibers for mechanical energy harvesting,” J. Nanotechnol. Eng. Med., vol. 5, no. 1, article no. 011002, 2014. 18 Volume 21 (1) 2022 Widhiyanuriyawan et al. Widhiyanuriyawan et al. 14. Z. Arifin, S. Hadi, S. Suyitno, A. R. Prabowo, and S. D. Prasetyo, “Characterization of zno nanofiber on
double-layer dye-sensitized solar cells using direct deposition method,” Period. Tche Quim., vol. 17, no. 36,
pp. 263-277, 2020. pp
15. H. N. Jati, M. Z. Khusaini, H. Sutanto, and Z. Arifin, “Application of direct deposition method for dye-
sensitized solar cell manufacturing process Application of Direct Deposition Method for Dye-Sensitized Solar
Cell Manufacturing Process,” AIP Conf. Proc., vol. 2097, article no. 030022, 2019. g
16. B. Sutanto, Z. Arifin, S. Hadi, L. M. Pranoto, and Y. V. Agustia, “Enhancement ZnO Nanofiber as
Semiconductor for Dye- Sensitized Solar Cells by Using Al Doped,” AIP Conf. Proc., vol. 1717, article no. 040006, 2016. 17. L. L. Tobin, T. O’Reilly, D. Zerulla, and J. T. Sheridan, “Characterising dye-sensitised solar cells,” Optik,
vol. 122, no. 14, pp. 1225-1230, 2011. pp
18. K. Sharma, V. Sharma, and S. S. Sharma, “Dye-Sensitized Solar Cells : Fundamentals and Current Status,”
Nanoscale Res. Lett., vol. 13, article no. 381, 2018. 19
|
https://openalex.org/W2981522273
|
https://jurnal.uns.ac.id/sepa/article/download/28152/pdf
|
Indonesian
| null |
ANALISIS PERWILAYAHAN DAN KONTRIBUSI KOMODITAS JERUK SIAM TERHADAP PEREKONOMIAN KABUPATEN BANYUWANGI
|
Sepa
| 2,019
|
cc-by
| 6,338
|
ISSN : 1829-9946 ISSN : 1829-9946 ISSN : 1829-9946 SEPA : Vol. 16 No.1 September 2019 : 85 – 96
DOI: https://doi.org/10.20961/sepa.v16i1.28152 ANALISIS PERWILAYAHAN DAN KONTRIBUSI KOMODITAS JERUK SIAM
TERHADAP PEREKONOMIAN KABUPATEN BANYUWANGI Rizqa Nurmalia*, Soetriono, Anik Suwandari
Program Studi Agribisnis, Fakultas Pertanian, Universitas Jember,
Jl. Kalimantan No. 37 Kampus Tegalboto Sumbersari Jember
*Corresponding author: nurmaliarizqa@gmail.com Abstract: Siam Oranges in Banyuwangi is spread in nine central districts. The aim of
the study is to explain the districts that are the basis of Siam Oranges, the
characteristics of the spread of Siam Oranges, and it contribution to the economy of
Banyuwangi. The method used in this study is an analytical method. The data used in
the study were secondary data from the Department of Agriculture, Forestry, and
Plantation of Banyuwangi. Analysis of data with the LQ method, localization and
specialization, and contributions will be done through the proportion formula. LQ
analysis is calculated using three indicators, namely land area, production and
revenue. The results of the analysis show that the based on the average LQ value from
2013-2017 which has consistently been the base district of the Siam Oranges based on
three different indicators are Bangorejo District, Purwoharjo District, Tegaldlimo
District, Cluring District, and Gambiran District. Localization analysis shows that
Siam Oranges are not localized in a particular district. Specialization analysis shows
that in none of the sub-districts of the Siam Oranges base specialize in just the Siam
Oranges commodity. Contribution analysis shows that the contribution of Siam
Oranges to the economy of Banyuwangi from 2013-2017 is 0,0019; 0,0041; 0,0102;
0,0050 dan 0,0020. It means low because these presentage is less than the average
contribution component of economy Banyuwangi. PENDAHULUAN yang menjadi daerah dengan produksi Jeruk
Siam tertinggi
serta beragamnya jumlah
produksi yang dihasilkan oleh masing-masing
kecamatan menunjukkan potensi yang berbeda
pula yang dimiliki tiap daerah. Pusat Data dan Informasi Pertanian (2016)
mencatat bahwa Jeruk Siam di Jawa Timur
dihasilkan dari beberapa kabupaten sebagai
sentra produksinya, diantaranya Kabupaten
Banyuwangi,
Jember,
Malang,
Pasuruan,
Lumajang dan lainnya. Produksi Jeruk Siam
tertinggi tahun 2015 sebesar 205.685 ton
dihasilkan oleh Kabupaten Banyuwangi dengan
nilai share terhadap provinsi sebesar 42,82%. Hal tersebut dapat diartikan bahwa hampir
sebagian besar kebutuhan Jeruk Siam Provinsi
Jawa Timur adalah dipenuhi oleh Kabupaten
Banyuwangi. Berikut gambaran perkembangan
produksi Jeruk Siam di beberapa kecamatan
penghasil
Jeruk
Siam
di
Kabupaten
Banyuwangi. Berdasarkan Rencana Strategis Dinas
Pertanian Kabupaten Banyuwangi Tahun 2016-
2021, salah satu strategi pembangunan yang
harus
diperhatikan
adalah
strategi
pengembangan kawasan strategis kabupaten
melalui
strategi
pengembangan
kawasan
strategis ekonomi yang meliputi penetapan
kawasan hortikultura. Mendukung strategi
tersebut,
produk
hortikultura
Kabupaten
Banyuwangi yang terbagi menjadi dua produk
yaitu sayuran dan buah-buahan dimana salah
satu komoditas hortikultura buah unggulannya
adalah Jeruk Siam yang tersebar di beberapa
kecamatan dengan produksi berbeda-beda juga
terlihat adanya gambaran pergantian kecamatan
yang menempati posisi sebagai kecamatan
dengan produksi Jeruk Siam tertinggi tiap
tahunnya, maka melalui gambaran tersebut
tentunya komoditas Jeruk Siam juga perlu
menjadi salah satu fokus komoditas yang
memerlukan penetapan kawasan strategisnya
sehingga dapat teridentifikasi dengan jelas
kecamatan mana yang memiliki keunggulan
komparatif yang potensial untuk dilakukan
pengembangan. Oleh karena itu, pemetaan
wilayah untuk komoditas Jeruk Siam di
Banyuwangi
menjadi
penting
dilakukan. Adapun tujuan dari penelitian adalah untuk Berdasarkan Tabel 1 terlihat bahwa
produksi Jeruk Siam yang dihasilkan dari tiap-
tiap kecamatan dari tahun 2013-2017 adalah
berbeda-beda. Tahun 2013 produksi Jeruk Siam
tertinggi
dihasilkan
oleh
Kecamatan
Tegaldlimo sebesar 395.718,75 kw. Produksi
Jeruk Siam tertinggi tahun 2014 dihasilkan oleh
Kecamatan Bangorejo sebesar 2.457.890 kw. Tahun 2015 Kecamatan Bangorejo masih
menjadi penghasil Jeruk Siam tertinggi yaitu
sebesar 8.039.430,00 kw. Tahun 2016 produksi
Jeruk Siam tertinggi juga masih dihasilkan oleh
Kecamatan Bangorejo dengan produksi sebesar
1.800.016 kw. Tahun 2017 produksi Jeruk
Siam
tertinggi
kembali
dihasilkan
oleh
Kecamatan Tegaldlimo. Pergantian Kecamatan Tabel 1. Perkembangan Produksi Jeruk Siam di Beberapa Kecamatan di Kabupaten Banyuwangi Tahun
2013-2017
No. Kecamatan
Produksi (kw)
2013
2014
2015
2016
2017
1. Pesanggaran
33.795,90
50.785,00
71.474,00
60.239,00
14.940,00
2. Siliragung
37.199,25
69.230,00
171.260,00
412.984,00
303.610,00
3. Bangorejo
381.037,50
2.457.890,00
8.039.430,00
1.800.016,00
113.488,00
4. Purwoharjo
347.568,30
290.565,00
106.841,00
449.654,00
177.791,00
5. Tegaldlimo
395.718,75
300.249,00
371.304,00
1.024.242,00
942.100,00
6. kontribusi komponen penyusun PDRB perekonomian Kabupaten Banyuwangi
(0,015%). Kata Kunci: jeruk siam, kecamatan basis, lokalisasi, spesialisasi, kontribusi. Keywords: siam oranges, base districts, localization, specialization, contribution. Abstrak: Jeruk Siam di Kabupaten Banyuwangi tersebar di sembilan kecamatan
sentra. Tujuan penelitian adalah untuk menjelaskan kecamatan basis Jeruk Siam,
karakteristik penyebarannya, serta kontribusinya terhadap perekonomian Kabupaten
Banyuwangi. Metode yang digunakan dalam penelitian adalah metode analitik. Data
yang digunakan dalam penelitian adalah data sekunder yang berasal dari Dinas
Pertanian, Kehutanan, dan Perkebunan Kabupaten Banyuwangi. Analisis data
menggunakan metode LQ, lokalisasi dan spesialisasi, serta analisis kontribusi
ekonomi melalui rumus proporsi. Analisis LQ dihitung dengan menggunakan tiga
indikator yaitu luas lahan, produksi dan penerimaan. Hasil penelitian menunjukkan
bahwa berdasarkan rata-rata nilai LQ tahun 2013-2017, yang secara konsisten menjadi
kecamatan basis Jeruk Siam pada tiga indikator yang berbeda selama kurun waktu
lima tahun yaitu Kecamatan Bangorejo, Kecamatan Purwoharjo, Kecamatan
Tegaldlimo, Kecamatan Cluring, dan Kecamatan Gambiran. Analisis lokalisasi
menunjukkan bahwa Jeruk Siam tidak terlokalisasi pada kecamatan tertentu melainkan
terlokalisasi di kecamatan-kecamatan basis. Analisis spesialisasi menunjukkan bahwa
Kabupaten Banyuwangi tidak menspesialisasikan kegiatan pertaniannya hanya pada
komoditas Jeruk Siam saja. Analisis kontribusi menunjukkan bahwa prsentase
kontribusi Jeruk Siam terhadap perekonomian Banyuwangi dari tahun 2013 sampai
2017 berturut-turut adalah 0,0019; 0,0041; 0,0102; 0,0050 dan 0,0020 yang berarti
bahwa kontribusi Jeruk Siam terhadap perekonomian Kabupaten Banyuwangi adalah
rendah karena angka presentase kontribusi Jeruk Siam lebih kecil dari rata-rata 85 Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … kontribusi komponen penyusun PDRB perekonomian Kabupaten Banyuwangi
(0,015%). PENDAHULUAN Muncar
62.295,75
30.810,00
0
191.280,00
123.760,00
7. Cluring
61.524,45
45.905,00
22.037,00
482.672,00
691.746,00
8. Gambiran
50.597,33
33.210,00
29.125,00
577.432,00
171.521,00
9. Tegalsari
67.770,00
31.865,00
0
151.630,00
67.456.00
Sumber: Dinas Pertanian Kehutanan dan Perkebunan Banyuwangi, 2018 (data diolah). embangan Produksi Jeruk Siam di Beberapa Kecamatan di Kabupaten Banyuwangi Tahun
3 2017 86 Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … menjelaskan kecamatan basis Jeruk Siam,
karakteristik penyebaran Jeruk Siam serta
kontribusi Jeruk Siam terhadap perekonomian
Kabupaten Banyuwangi. (β=1,677), dan nanas (β=1,440). Artinya
keenam komoditi buah tersebut dapat dijadikan
andalan hortikultura buah-buahan Provinsi
Gorontalo yang dapat dikembangkan mulai dari
agribisnis hulu sampai hilir karena memiliki
keunggulan komparatif. Berdasarkan hasil penelitian Mulyono
dan Munibah (2016) yang menggunakan
pendekatan Location Quotient dan Shift Share
Analysis dalam penentuan komoditas unggulan
di Kabupaten Bantul, diketahui bahwa Padi
Sawah memiliki keunggulan komparatif di 12
kecamatan dengan nilai LQ terbesar yaitu 1,33,
Jagung memiliki keunggulan komparatif di 4
kecamatan dengan nilai LQ terbesar yaitu 2,29,
Kacang
Tanah
memiliki
keunggulan
komparatif di 6 kecamatan dengan nilai LQ
terbesar yaitu 3,10, Ubi Jalar memiliki
keunggulan komparatif di 3 kecamatan dengan
nilai LQ terbesar yaitu 18,04, serta Ubi Kayu
memiliki
keunggulan
komparatif
di
3
kecamatan dengan nilai LQ terbesar yaitu 4,14. Komoditas-komoditas
tersebut
memiliki
keunggulan komparatif di kecamatan tertentu
karena
share
luas
panennya
di
tingkat
kecamatan lebih tinggi dibandingkan dengan
tingkat kabupaten, sehingga nilai LQ lebih
besar dari 1. Komoditas yang memiliki
keunggulan
komparatif
menunjukkan
komoditas
tersebut
diproduksi
melalui
dominasi dukungan sumber daya alam, dimana
daerah lain tidak mampu memproduksinya. gg
p
Larasati (2011) melakukan penelitian
untuk mengetahui kontribusi ubi jalar terhadap
PDRB Provinsi Jawa Timur menggunakan
analisis
proporsi
dengan
kriteria
yang
digunakan adalah kontribusi ubi jalar dikatakan
tinggi jika presentasenya lebih dari 0,093% dan
dikatakan rendah jika presentasenya kurang
dari 0,093%. Berdasarkan hasil penelitian,
diketahui bahwa pada tahun 1994 kontribusi
ubi jalar sebesar 0,0025% dan mengalami
peningkatan
pada
tahun
1995
menjadi
0,0030%. Kontribusi ubi jalar pada tahun 1996
dan
1997
adalah
0,002%
dan
terjadi
peningkatan pada tahun 1998 yaitu 0,005%. Pada tahun 1999 kontribusinya mengalami
penurunan yaitu sebesar 0,001% dan semakin
menurun pada tahun 2000 sampai dengan 2009
menjadi
dibawah
0,001%. Hal
tersebut
menunjukkan bahwa kontribusi ubi jalar
terhadap PDRB Provinsi Jawa Timur dari tahun
1994-2009 cenderung mengalami penurunan
dan secara keseluruhan kontribusinya dikatakan
rendah karena kurang dari 0,093%. PENDAHULUAN Penelitian analisis perwilayahan dan
kontribusi Jeruk Siam di Banyuwangi penting
dilakukan melihat kondisi tersebarnya Jeruk
Siam di beberapa wilayah sentra dengan
produksi
yang
berbeda-beda. Gambaran
tersebut menunjukkan bahwa potensi yang
dimiliki tiap wilayahpun berbeda. Wilayah
basis Jeruk Siam adalah wilayah yang memiliki
keunggulan komparatif dibandingkan wilayah
non basis Jeruk Siam sehingga wilayah basis
merupakan wilayah yang paling potensial untuk
dilakukan
pengembangan. Kebaruan
dari
penelitian
ini
adalah
apabila
penelitian-
penelitian sebelumnya hanya melibatkan 1
indikator saja dalam menentukan wilayah basis
namun
pada
penelitian
ini
dilakukan
perhitungan LQ dengan menggunakan 3
indikator. Hal tersebut dapat menunjukkan
wilayah yang mana yang secara konsisten
menjadi wilayah basis Jeruk Siam dengan tiga
indikator berbeda sehingga wilayah basis Jeruk
Siam yang dihasilkan dalam penelitian terbukti
benar-benar unggul tidak hanya dalam satu atau
dua indikator namun unggul dalam tiga Boekoesoe (2011) dengan menggunakan
analisis LQ, menemukan bahwa terdapat 18
kecamatan
yang
menjadi
basis
wilayah
komoditas mangga dimana luas lahan tanaman
mangga yang terbesar adalah di Kecamatan
Tilongkabila,
Kabupaten
Bone
Bolango. Terdapat 19 komoditi buah-buahan di Provinsi
Gorontalo. Berdasarkan analisis lokalisasi
terdapat 3 komoditi buah-buahan yang nilai
koefisien lokalisasinya lebih besar dari satu
(α>1), yaitu jeruk besar (α=1,491), rambutan
(α=1,327), dan salak (α=1,258). Sedangkan
komoditi yang paling kecil nilai koefisien
lokalisasinya adalah pisang (α=0,175), yang
berarti bahwa komoditi pisang adalah paling
menyebar di wilayah-wilayah kecamatan di
Provinsi Gorontalo atau dengan kata lain
hampir seluruh kecamatan terdapat pisang. Berdasarkan analisis koefisien spesialisasi
terdapat 6 komoditi buah yang menjadi spesial
bagi Provinsi Gorontalo dengan nilai β>1 yaitu
mangga (β=4,178), nangka (β=2,912), pisang
(β=2,907), pepaya (β=1,940), jeruk siam 87 Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Vi
=
Luas lahan, produksi, penerimaan
komoditas Jeruk Siam periode
tahun 2013-2017 di Kabupaten
Banyuwangi indikator. Analisis kontribusi dilakukan untuk
menjelaskan Jeruk Siam sebagai salah satu
komoditas buah unggulan yang produksinya
tinggi di Kabupaten Banyuwangi apakah dapat
memberikan kontribusi yang tinggi pula
terhadap perekonomian Banyuwangi. y
g
Vt
=
Total
luas
lahan,
produksi,
penerimaan
seluruh
komoditas
buah periode tahun 2013-2017 di
Kabupaten Banyuwangi METODE PENELITIAN Penelitian dilakukan di Kabupaten Banyuwangi
Provinsi Jawa Timur. Metode yang digunakan
dalam penentuan daerah penelitian adalah
metode secara sengaja (purposive methode). Pemilihan daerah penelitian didasarkan atas
pertimbangan bahwa Kabupaten Banyuwangi
merupakan wilayah dengan tingkat kontribusi
tertinggi Jeruk Siam di Provinsi Jawa Timur,
Metode yang digunakan dalam penelitian ini
adalah metode analitik. Menurut Nazir (2005),
metode analitik bertujuan untuk menguji
hipotesis-hipotesis dan melakukan interpretasi
yang lebih mendalam tentang hubungan-
hubungan. Data
yang
digunakan
dalam
penelitian adalah data sekunder. Sumber data
utama dalam penelitian berasal dari Dinas
Pertanian,
Kehutanan,
dan
Perkebunan
Kabupaten Banyuwangi, Badan Pusat Statistik
(BPS) Kabupaten Banyuwangi, serta instansi
terkait lainnya. Adapun data yang diperlukan
dalam penelitian adalah data luas lahan,
produksi,
dan
harga
buah-buahan
setiap
kecamatan di Kabupaten Banyuwangi tahun
2013 sampai dengan 2017 serta data PDRB
Kabupaten Banyuwangi atas dasar harga
berlaku tahun 2013-2017. Kriteria pengambilan keputusan: 1. Apabila LQ >1, artinya kecamatan i
merupakan basis Jeruk Siam berdasarkan
indikator luas lahan, produksi, penerimaan. 2. Apabila LQ<1, artinya kecamatan i bukan
merupakan basis Jeruk Siam berdasarkan
indikator luas lahan, produksi, penerimaan. Analisis data yang digunakan untuk
mencapai tujuan kedua menggunakan analisis
lokalisasi dan spesialisasi. Menurut Pasaribu
dan Soetriono (2009) menyatakan bahwa
penggunaan analisis lokalisasi dan spesialisasi
dapat digunakan rumus: Rumus Lokalisasi: αi= [Si/Ni] – [∑Si/∑Ni] ...(2) Rumus Lokalisasi: αi= [Si/Ni] – [∑Si/∑Ni] ...(2) αi
=
Koefisien
lokalisasi
komoditas
Jeruk Siam Si
=
Luas lahan, produksi, penerimaan
komoditas Jeruk Siam periode
tahun 2013-2017 di Kecamatan i Ni
=
Luas lahan, produksi, penerimaan
komoditas Jeruk Siam periode
tahun 2013-2017 di Kabupaten
Banyuwangi Analisis data yang digunakan untuk
mencapai tujuan pertama menggunakan analisis
LQ (Location Quotient) oleh Soetriono (1996)
dan Suharyanto et al. (2013) dengan rumus: y
g
∑Si
=
Total
luas
lahan,
produksi,
penerimaan
seluruh
komoditas
buah periode tahun 2013-2017 di
Kecamatan i LQ = (vi/vt)/(Vi/Vt) ……………………… ( 1 )
Keterangan:
LQ
=
Nilai LQ rasio komoditas Jeruk
Siam di setiap kecamatan di
Kabupaten Banyuwangi
vi
=
Luas lahan, produksi, penerimaan
komoditas Jeruk Siam periode
tahun 2013-2017 di Kecamatan i
vt
=
Total
Luas
lahan,
produksi,
penerimaan
seluruh
komoditas
buah periode tahun 2013-2017 di
Kecamatan i LQ = (vi/vt)/(Vi/Vt) ……………………… ( 1 ) ∑Ni
=
Total
luas
lahan,
produksi,
penerimaan
seluruh
komoditas
buah periode tahun 2013-2017 di
Kabupaten Banyuwangi g
LQ
=
Nilai LQ rasio komoditas Jeruk
Siam di setiap kecamatan di
Kabupaten Banyuwangi Kriteria pengambilan keputusan: Kriteria pengambilan keputusan: vi
=
Luas lahan, produksi, penerimaan
komoditas Jeruk Siam periode
tahun 2013-2017 di Kecamatan i 1. Apabila α≥1, artinya komoditas Jeruk Siam
terlokalisasi pada suatu kecamatan tertentu. 2. Apabila α<1, artinya komoditas Jeruk Siam
tidak terlokalisasi pada suatu kecamatan
tertentu atau dengan kata lain menyebar
dibeberapa
kecamatan
di
Kabupaten
Banyuwangi. vt
=
Total
Luas
lahan,
produksi,
penerimaan
seluruh
komoditas
buah periode tahun 2013-2017 di
Kecamatan i 88 Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … 2. Rumus Lokalisasi: αi= [Si/Ni] – [∑Si/∑Ni] ...(2) Apabila kontribusi (%) rata-rata kontribusi
PDRB
Kabupaten
Banyuwangi,
maka
kontribusi Jeruk Siam terhadap PDRB
Kabupaten Banyuwangi adalah rendah Keterangan: g
βi
=
Koefisien spesialisasi komoditas
Jeruk Siam Si
=
Luas lahan, produksi, penerimaan
komoditas Jeruk Siam periode
tahun 2013-2017 di Kecamatan i Ni
=
Luas lahan, produksi, penerimaan
komoditas Jeruk Siam periode
tahun 2013-2017 di Kabupaten
Banyuwangi ∑ komponen penyusun PDRB Kabupaten
Banyuwangi = sektor pertanian, kehutanan, dan
perikanan;
sektor
pertambangan
dan
penggalian; sektor industri pengolahan; sektor
pengadaan listrik dan gas; sektor pengadaan air,
pengelolaan sampah, limbah, dan daur ulang;
sektor konstruksi; sektor perdagangan besar
dan eceran, reparasi mobil dan sepeda motor;
sektor transportasi dan pergudangan; sektor
penyediaan akomodasi dan makan minum;
sektor informasi dan komunikasi; sektor jasa
keuangan dan asuransi; sektor real estate;
sektor jasa perusahaan; sektor administrasi
pemerintahan, pertahanan dan jaminan sosial
wajib; sektor jasa pendidikan; sektor jasa
kesehatan dan kegiatan sosial; serta sektor jasa
lainnya. y
g
∑Si
=
Total
luas
lahan,
produksi,
penerimaan
seluruh
komoditas
buah periode tahun 2013-2017 di
Kecamatan i ∑Ni
=
Total
luas
lahan,
produksi,
penerimaan
seluruh
komoditas
buah periode tahun 2013-2017 di
Kabupaten Banyuwangi Kriteria pengambilan keputusan: Kriteria pengambilan keputusan: Kriteria pengambilan keputusan: Kriteria pengambilan keputusan: 1. Apabila
β 1,
artinya
suatu
wilayah
(kecamatan) sudah menspesialisasikan pada
satu jenis komoditas pertanian tertentu
dalam hal ini komoditas Jeruk Siam. 2. Apabila
β<1,
artinya
suatu
wilayah
(kecamatan)
belum
dan
atau
tidak
menspesialisasikan
pada
satu
jenis
komoditas pertanian tertentu dalam hal ini
komoditas Jeruk Siam. Nilai rata-rata kontribusi diatas masih
dibagi lagi dengan komponen penyusun sektor
pertanian yang terdiri dari 7 subsektor,
sehingga perhitungannya menjadi: Rata-rata
kontribusi
komponen
penyusun
Sektor pertanian = 4,76% / 7 = 0,68% Analisis data yang digunakan untuk
mencapai tujuan ketiga menggunakan rumus
proporsi
oleh
Djarwanto
(2001)
sebagai
berikut: Nilai rata-rata kontribusi diatas masih
harus dibagi lagi dengan total komoditas
penyusun subsektor hortikultura yang terdiri
dari 48 komoditas, sehingga perhitungannya
menjadi: Kontribusi (%) = [X/Y] x 100% Kontribusi (%) = [X/Y] x 100% Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … 1. Apabila kontribusi (%) > 0,015%, maka
kontribusi komoditas Jeruk Siam terhadap
PDRB
Kabupaten
Banyuwangi
adalah
tinggi. 1. Apabila kontribusi (%) > 0,015%, maka
kontribusi komoditas Jeruk Siam terhadap
PDRB
Kabupaten
Banyuwangi
adalah
tinggi. Kecamatan
Tegaldlimo,
Kecamatan
Bangorejo, Kecamatan Muncar dan Kecamatan
Pesanggaran
tidak
menjadi
kecamatan
konsisten karena selama kurun waktu 2013-
2017 terdapat satu tahun tertentu pada
kecamatan-kecamatan
tersebut
yang
menghasilkan nilai LQ kurang dari 1 atau tidak
menjadi kecamatan basis Jeruk Siam. Hasil
terebut secara berturut-turut yaitu Kecamatan
Tegaldlimo menjadi kecamatan non basis Jeruk
Siam pada tahun 2014, Kecamatan Bangorejo
menjadi kecamatan non basis Jeruk Siam pada
tahun 2017, Kecamatan Muncar menjadi
kecamatan non basis Jeruk Siam pada tahun
2014 dan 2015, Kecamatan Pesanggaran
menjadi kecamatan non basis Jeruk Siam pada
tahun 2015. Kecamatan-kecamatan tersebut
disebut non basis karena jumlah luas lahan
Jeruk Siam yang dimiliki tidak mampu
mengimbangi
total
luas
lahan
seluruh
komoditas buah yang diusahakan masyarakat
pada kecamatan-kecamatan tersebut yang juga
berarti bahwa ada komoditas buah lain yang
lebih mendominasi diusahakan masyarakat
pada kecamatan-kecamatan tersebut. 2. Apabila kontribusi (%) ≤ 0,015%, maka
kontribusi komoditas Jeruk Siam terhadap
PDRB
Kabupaten
Banyuwangi
adalah
rendah. Keterangan: Keterangan: Rata-rata
kontribusi
komponen
penyusun
PDRB total Kabupaten Banyuwangi =
0,68% / 48 = 0,015% X
= PDRB komoditas Jeruk Siam Kabupaten
Banyuwangi (Rp) Y
= PDRB sektor pertanian di Kabupaten
Banyuwangi (Rp) Berdasarkan
perhitungan
tersebut,
kriteria yang digunakan dalam menentukan
tinggi
rendahnya
kontribusi
Jeruk
Siam
terhadap PDRB Kabupaten Banyuwangi adalah
sebagai berikut: Kriteria pengambilan keputusan: Kriteria pengambilan keputusan: Kriteria pengambilan keputusan: 1. Apabila kontribusi (%) > rata-rata kontribusi
PDRB
Kabupaten
Banyuwangi,
maka
kontribusi Jeruk Siam terhadap PDRB
Kabupaten Banyuwangi adalah tinggi 89 Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … berdasarkan indikator produksi adalah tidak
terdapat kecamatan yang selalu menghasilkan
nilai LQ lebih dari satu (LQ>1) atau selalu
menjadi kecamatan basis Jeruk Siam selama
kurun waktu lima tahun tersebut. Kecamatan
Purwoharjo dengan rata-rata LQ tertinggi
pertama sebesar 1,75 tidak menjadi kecamatan
konsisten
karena
pada
tahun
2017
menghasilkan nilai LQ kurang dari 1 (LQ<1)
atau tidak menjadi kecamatan basis Jeruk Siam. Kecamatan Tegaldlimo dengan rata-rata LQ
tertinggi kedua sebesar 1,71 tidak menjadi
kecamatan konsisten karena pada tahun 2015
dan 2017 menjadi kecamatan non basis Jeruk
Siam. Kecamatan Bangorejo dengan rata-rata
LQ tertinggi ketiga sebesar 1,70 tidak menjadi
kecamatan konsisten karena pada tahun 2017
menjadi kecamatan non basis Jeruk Siam. Kecamatan Cluring dengan rata-rata LQ
tertinggi keempat sebesar 1,40 juga tidak
menjadi kecamatan konsisten karena pada
tahun 2014 dan 2015 menjadi kecamatan non
basis Jeruk Siam. Kecamatan Gambiran dengan
rata-rata LQ tertinggi kelima atau rata-rata LQ
terendah sebesar 1,39 tidak menjadi kecamatan
konsisten karena pada tahun 2014 menjadi
kecamatan non basis Jeruk Siam. Kecamatan-
kecamatan tersebut menjadi kecamatan non
basis Jeruk Siam karena jumlah produksi Jeruk
Siam yang dihasilkan pada tahun-tahun tersebut
tidak mampu mengimbangi total produksi
seluruh komoditas buah yang diusahakan
masyarakat
di
masing-masing
kecamatan. Artinya
komoditas
Jeruk
Siam
bukan
merupakan komoditas buah yang mendominasi
diusahakan di kecamatan-kecamatan tersebut. berdasarkan indikator produksi adalah tidak
terdapat kecamatan yang selalu menghasilkan
nilai LQ lebih dari satu (LQ>1) atau selalu
menjadi kecamatan basis Jeruk Siam selama
kurun waktu lima tahun tersebut. Kecamatan
Purwoharjo dengan rata-rata LQ tertinggi
pertama sebesar 1,75 tidak menjadi kecamatan
konsisten
karena
pada
tahun
2017
menghasilkan nilai LQ kurang dari 1 (LQ<1)
atau tidak menjadi kecamatan basis Jeruk Siam. Kecamatan Tegaldlimo dengan rata-rata LQ
tertinggi kedua sebesar 1,71 tidak menjadi
kecamatan konsisten karena pada tahun 2015
dan 2017 menjadi kecamatan non basis Jeruk
Siam. Kecamatan Bangorejo dengan rata-rata
LQ tertinggi ketiga sebesar 1,70 tidak menjadi
kecamatan konsisten karena pada tahun 2017
menjadi kecamatan non basis Jeruk Siam. Kecamatan Cluring dengan rata-rata LQ
tertinggi keempat sebesar 1,40 juga tidak
menjadi kecamatan konsisten karena pada
tahun 2014 dan 2015 menjadi kecamatan non
basis Jeruk Siam. Kecamatan Gambiran dengan
rata-rata LQ tertinggi kelima atau rata-rata LQ
terendah sebesar 1,39 tidak menjadi kecamatan
konsisten karena pada tahun 2014 menjadi
kecamatan non basis Jeruk Siam. HASIL DAN PEMBAHASAN Kecamatan Basis dan Non Basis Komoditas
Jeruk Siam di Kabupaten Banyuwangi
Berdasarkan analisis LQ indikator luas lahan
yang telah dilakukan, diperoleh nilai LQ Jeruk
Siam seperti yang tersaji pada Tabel 2. Kecamatan Basis dan Non Basis Komoditas
Jeruk Siam di Kabupaten Banyuwangi p
y
g
Berdasarkan analisis LQ indikator luas lahan
yang telah dilakukan, diperoleh nilai LQ Jeruk
Siam seperti yang tersaji pada Tabel 2. p
y
g
j p
Tabel 2. menunjukkan bahwa kecamatan
yang menjadi basis Jeruk Siam dari tahun
2013-2017
berubah-ubah. Hal
tersebut
disebabkan oleh luas lahan yang juga berbeda-
beda
setiap
tahunnya
di
masing-masing
kecamatan penghasil Jeruk Siam. Berdasarkan
rata-rata nilai LQ terdapat sembilan (9)
kecamatan yang merupakan wilayah basis
Jeruk Siam. Kesembilan kecamatan tersebut
adalah Kecamatan Pesanggaran, Siliragung,
Bangorejo, Purwoharjo, Tegaldlimo, Muncar,
Cluring, Gambiran, dan Kecamatan Tegalsari. Artinya bahwa komoditas Jeruk Siam di
Kabupaten Banyuwangi memiliki keunggulan
komparatif di sembilan kecamatan tersebut. Berdasarkan
analisis
LQ
indikator
produksi yang telah dilakukan, diperoleh hasil
analisis seperti yang tersaji pada Tabel 3. Hasil
analisis LQ berdasarkan indikator produksi
menunjukkan
bahwa
terdapat
lima
(5)
kecamatan yang menghasilkan nilai rata-rata
LQ > 1 atau disebut sebagai kecamatan basis
Jeruk Siam. Kelima kecamatan tersebut adalah
Kecamatan
Bangorejo,
Purwoharjo,
Tegaldlimo,
Cluring,
dan
Kecamatan
Gambiran. Berdasarkan konsistensinya per
tahun, yang secara konsisten menempati posisi
sebagai kecamatan basis komoditas Jeruk Siam Berdasarkan konsistensinya, yang secara
konsisten menempati posisi sebagai kecamatan
basis pada tahun 2013-2017 berdasarkan
indikator
luas
lahan
adalah
Kecamatan
Siliragung, Purwoharjo, Cluring, Gambiran,
dan Tegalsari. Tabel 2. Hasil analisis LQ Jeruk Siam Berdasarkan Indikator Luas Lahan (ha) di Kabupaten Banyuwangi
No
Kecamatan
Nilai LQ Indikator Luas Lahan
2013
2014
2015
2016
2017
Rata-rata
1
Pesanggaran
1,8508
1,6409
0,5458
1,1229
1,2448
1,2811
2
Siliragung
1,6741
2,2189
1,2737
1,3759
1,5289
1,6143
3
Bangorejo
1,6200
3,1763
2,4453
1,7960
0,0058
1,8087
4
Purwoharjo
4,0067
4,0564
2,7449
2,0951
2,2850
3,0376
5
Tegaldlimo
2,1316
0,7320
2,5638
2,0999
2,5999
2,0254
6
Muncar
2,1591
0,4577
0,6715
1,4938
1,6792
1,2922
7
Cluring
3,8228
1,4356
2,1609
1,9671
2,2000
2,3173
8
Gambiran
1,6786
1,0271
2,0795
1,6967
1,9256
1,6815
9
Tegalsari
3,9280
1,4089
1,3087
1,4856
1,6463
1,9555
Sumber: Data Sekunder Diolah (2018) isis LQ Jeruk Siam Berdasarkan Indikator Luas Lahan (ha) di Kabupaten Banyuwangi Tabel 2. Hasil analisis LQ Jeruk Siam Berdasarkan Indikator Luas Lahan (ha) di Kabupaten Ba 90 Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Kecamatan-
kecamatan tersebut menjadi kecamatan non
basis Jeruk Siam karena jumlah produksi Jeruk
Siam yang dihasilkan pada tahun-tahun tersebut
tidak mampu mengimbangi total produksi
seluruh komoditas buah yang diusahakan
masyarakat
di
masing-masing
kecamatan. Artinya
komoditas
Jeruk
Siam
bukan
merupakan komoditas buah yang mendominasi
diusahakan di kecamatan-kecamatan tersebut. Terdapat empat (4) kecamatan yang
sebelumnya telah dijelaskan pada hasil analisis
LQ
berdasarkan
indikator
luas
lahan
menghasilkan rata-rata nilai LQ lebih dari 1
atau
masuk
kedalam
kecamatan
basis
komoditas Jeruk Siam namun berdasarkan
indikator produksi tidak termasuk dalam
kecamatan
basis
Jeruk
Siam. Keempat
kecamatan
tersebut
adalah
Kecamatan
Pesanggaran,
Siliragung,
Muncar,
dan
Tegalsari. Empat kecamatan tersebut tidak
menjadi kecamatan basis dikarenakan hasil
rataan produksi Jeruk Siam di kecamatan-
kecamatan tersebut lebih rendah atau tidak
mampu mengimbangi hasil rataan produksi dari
seluruh komoditas buah dalam satu wilayah
kecamatan. Berdasarkan
analisis
LQ
indikator
penerimaan yang telah dilakukan, diperoleh
hasil seperti yang tersaji pada Tabel 4. Hasil
analisis LQ berdasarkan indikator penerimaan
menunjukkan
bahwa
terdapat
enam
(6)
kecamatan yang menghasilkan nilai rata-rata
LQ>1 atau disebut sebagai kecamatan basis
komoditas Jeruk Siam. Keenam kecamatan
tersebut
adalah
Kecamatan
Bangorejo,
Purwoharjo, Tegaldlimo, Cluring, Gambiran
dan Tegalsari. Berdasarkan konsistensi per tahun, yang
secara konsisten menempati posisi sebagai
kecamatan
basis
komoditas
Jeruk
Siam
berdasarkan indikator penerimaan adalah tidak
terdapat kecamatan yang secara konsisten
selalu menjadi kecamatan basis selama kurun
waktu
lima
tahun
tersebut. Kecamatan
Tegaldlimo dengan rata-rata LQ tertinggi Tabel 3. Hasil Analisis LQ Jeruk Siam Berdasarkan Indikator Produksi (kw) di Kabupaten Banyuwangi
No
Kecamatan
Nilai LQ Indikator Produksi
2013
2014
2015
2016
2017
Rata-rata
1
Pesanggaran
0,5974
0,3545
0,5541
0,2791
0,0953
0,3761
2
Siliragung
0,6676
0,2433
0,0937
1,3709
1,3947
0,7541
3
Bangorejo
2,1263
1,9773
1,5497
2,0874
0,7451
1,6971
4
Purwoharjo
3,0693
1,8377
1,1603
1,8755
0,8022
1,7490
5
Tegaldlimo
2,5910
1,1645
0,8386
1,9032
2,0406
1,7076
6
Muncar
0,6436
0,1957
0
0,8676
1,3382
0,6090
7
Cluring
1,4871
0,9755
0,4648
2,0553
2,0288
1,4023
8
Gambiran
1,2089
0,7387
1,2046
2,0023
1,8307
1,3970
9
Tegalsari
1,4227
0,7119
0
1,5723
1,2587
0,9931
Sumber: Data Sekunder Diolah (2018) Q Jeruk Siam Berdasarkan Indikator Produksi (kw) di Kabupaten Banyuwangi Tabel 3. Hasil Analisis LQ Jeruk Siam Berdasarkan Indikator Produksi (kw) di Kabupaten Banyuwangi 91 Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … sebesar
1,57
tidak
menjadi
kecamatan
konsisten
karena
pada
tahun
2015
menghasilkan nilai LQ kurang dari 1 (LQ<1)
atau tidak menjadi kecamatan basis Jeruk Siam. Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Kecamatan Bangorejo dengan rata-rata LQ
tertinggi kedua sebesar 1,38 tidak menjadi
kecamatan konsisten karena pada tahun 2017
menjadi kecamatan non basis. Kecamatan
Purwoharjo dengan rata-rata LQ tertinggi
ketiga sebesar 1,37 tidak menjadi kecamatan
konsisten karena pada tahun 2015 dan 2017
menjadi kecamatan non basis. Kecamatan
Cluring dengan rata-rata LQ tertinggi keempat
sebesar
1,35
tidak
menjadi
kecamatan
konsisten karena pada tahun 2013 dan 2014
menjadi kecamatan non basis. Kecamatan
Gambiran dengan rata-rata LQ tertinggi kelima
sebesar
1,31
tidak
menjadi
kecamatan
konsisten karena pada tahun 2014 menjadi kecamatan non basis. Kecamatan Tegalsari
dengan rata-rata LQ tertinggi keenam atau rata-
rata LQ terendah sebesar 1,11 tidak menjadi
kecamatan konsisten karena pada tahun 2014
dan 2015 menjadi kecamatan non basis. Kecamatan non basis Jeruk Siam berdasarkan
indikator penerimaan dapat diartikan bahwa
Jeruk
Siam
bukan
merupakan
sumber
penerimaan terbesar bagi kecamatan-kecamatan
tersebut. Rangkuman kecamatan yang menjadi
basis Jeruk Siam berdasarkan rata-rata nilai LQ
dalam tiga indikator berbeda yaitu luas lahan,
produksi, dan penerimaan tersaji pada Tabel 5. Berdasarkan Tabel 5. diketahui bahwa
dalam tiga indikator berbeda terdapat jumlah
yang berbeda pula yang menjadi kecamatan
basis Jeruk Siam di Kabupaten Banyuwangi. Tabel 4. Hasil Analisis LQ Jeruk Siam Berdasarkan Indikator Penerimaan (Rp) di Kabupaten Banyuwangi
No
Kecamatan
Nilai LQ Indikator Penerimaan
2013
2014
2015
2016
2017
Rata-rata
1
Pesanggaran
0,7134
0,4932
0,5577
0,3720
0,0717
0,4416
2
Siliragung
0,7596
0,2715
0,1031
0,8121
1,5138
0,6920
3
Bangorejo
1,8820
1,5888
1,3519
1,5426
0,5463
1,3823
4
Purwoharjo
2,2620
1,4371
0,9520
1,4879
0,7517
1,3781
5
Tegaldlimo
2,0787
1,0899
0,9809
1,3780
2,3132
1,5681
6
Muncar
0,6915
0,2653
0
0,8598
1,9659
0,7565
7
Cluring
1,4880
0,9783
0,5780
1,5404
2,1803
1,3530
8
Gambiran
1,1756
0,7124
1,1036
1,5047
2,0611
1,3115
9
Tegalsari
1,5894
0,9108
0
1,3152
1,7157
1,1062
Sumber: Data Sekunder Diolah (2018)
Tabel 5. Kecamatan Basis Jeruk Siam di Kabupaten Banyuwangi Indikator Luas Lahan (ha), Produksi (kw)
dan Penerimaan (rp)
No. Indikator
Luas Lahan
Produksi
Penerimaan
1. Purwoharjo
Purwoharjo
Tegaldlimo
2. Cluring
Tegaldlimo
Bangorejo
3. Tegaldlimo
Bangorejo
Purwoharjo
4. Tegalsari
Cluring
Cluring
5. Bangorejo
Gambiran
Gambiran
6. Gambiran
Tegalsari
7. Siliragung
8. Muncar
9. Pesanggaran
Sumber: Data Sekunder Diolah (2018) Tabel 4. Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … (2016)
berdasarkan
perhitungan koefisien lokalisasi, diketahui
bahwa tidak ada yang mencapai nilai 1 artinya
bahwa Kabupaten Cirebon tidak melakukan
konsentrasi kegiatan usahatani atau kegiatan
usahataninya tidak terkumpul hanya di salah
satu kecamatan melainkan tersebar di beberapa
kecamatan di Kabupaten Cirebon. Selama
kurun waktu tahun 2013 hingga 2017 kegiatan
pertanian
komoditas
Jeruk
Siam
tidak
terlokalisasi atau menyebar. kecamatan
yang
bersangkutan
memiliki
karakteristik iklim, cuaca, dan topografi yang
hampir sama sehingga Jeruk Siam dapat
dikembangkan dengan baik di kecamatan-
kecamatan
tersebut. Tersebarnya
kegiatan
usaha pertanian Jeruk Siam dapat pula
memberikan keuntungan bagi pelaku usaha-
usaha yang berkaitan dengan komoditas Jeruk
Siam karena apabila kebutuhan terhadap Jeruk
Siam tidak bisa diperoleh di suatu kecamatan
maka masih ada kecamatan-kecamatan lain
yang dapat dijadikan rujukan untuk memenuhi
kebutuhan akan Jeruk Siam. Hal ini sesuai
dengan
penelitian
yang
telah
dilakukan
Syamsiyah
et
al. (2016)
berdasarkan
perhitungan koefisien lokalisasi, diketahui
bahwa tidak ada yang mencapai nilai 1 artinya
bahwa Kabupaten Cirebon tidak melakukan
konsentrasi kegiatan usahatani atau kegiatan
usahataninya tidak terkumpul hanya di salah
satu kecamatan melainkan tersebar di beberapa
kecamatan di Kabupaten Cirebon. Selama
kurun waktu tahun 2013 hingga 2017 kegiatan
pertanian
komoditas
Jeruk
Siam
tidak
terlokalisasi atau menyebar. Berdasarkan rata-rata nilai LQ tahun 2013
sampai 2017 yang secara konsisten selalu
menjadi kecamatan basis Jeruk Siam pada 3
indikator yang berbeda adalah Kecamatan
Bangorejo, Kecamatan Purwoharjo, Kecamatan
Tegaldlimo,
Kecamatan
Cluring,
dan
Kecamatan
Gambiran. Artinya
bahwa
kecamatan-kecamatan
basis
tersebut
merupakan
wilayah
yang
relatif
lebih
berpotensi dilakukan pengembangan daripada
kecamatan basis yang nonkonsisten. Berdasarkan rata-rata nilai LQ tahun 2013
sampai 2017 yang secara konsisten selalu
menjadi kecamatan basis Jeruk Siam pada 3
indikator yang berbeda adalah Kecamatan
Bangorejo, Kecamatan Purwoharjo, Kecamatan
Tegaldlimo,
Kecamatan
Cluring,
dan
Kecamatan
Gambiran. Artinya
bahwa
kecamatan-kecamatan
basis
tersebut
merupakan
wilayah
yang
relatif
lebih
berpotensi dilakukan pengembangan daripada
kecamatan basis yang nonkonsisten. Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Hasil Analisis LQ Jeruk Siam Berdasarkan Indikator Penerimaan (Rp) di Kabupaten Banyuwangi
No
Kecamatan
Nilai LQ Indikator Penerimaan
2013
2014
2015
2016
2017
Rata-rata
1
Pesanggaran
0,7134
0,4932
0,5577
0,3720
0,0717
0,4416
2
Siliragung
0,7596
0,2715
0,1031
0,8121
1,5138
0,6920
3
Bangorejo
1,8820
1,5888
1,3519
1,5426
0,5463
1,3823
4
Purwoharjo
2,2620
1,4371
0,9520
1,4879
0,7517
1,3781
5
Tegaldlimo
2,0787
1,0899
0,9809
1,3780
2,3132
1,5681
6
Muncar
0,6915
0,2653
0
0,8598
1,9659
0,7565
7
Cluring
1,4880
0,9783
0,5780
1,5404
2,1803
1,3530
8
Gambiran
1,1756
0,7124
1,1036
1,5047
2,0611
1,3115
9
Tegalsari
1,5894
0,9108
0
1,3152
1,7157
1,1062
Sumber: Data Sekunder Diolah (2018) Hasil Analisis LQ Jeruk Siam Berdasarkan Indikator Penerimaan (Rp) di Kabupaten Banyuwangi
Nilai LQ Indikator Penerimaan Tabel 5. Kecamatan Basis Jeruk Siam di Kabupaten Banyuwangi Indikator Luas Lahan (ha), Produksi (kw)
dan Penerimaan (rp)
No. Indikator
Luas Lahan
Produksi
Penerimaan
1. Purwoharjo
Purwoharjo
Tegaldlimo
2. Cluring
Tegaldlimo
Bangorejo
3. Tegaldlimo
Bangorejo
Purwoharjo
4. Tegalsari
Cluring
Cluring
5. Bangorejo
Gambiran
Gambiran
6. Gambiran
Tegalsari
7. Siliragung
8. Muncar
9. Pesanggaran
Sumber: Data Sekunder Diolah (2018) 92 Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Tabel 6. Hasil Analisis Lokalisasi Jeruk Siam Berdasarkan Indikator Luas Lahan, Produksi dan Penerimaan
Tahun 2013-2017 di Kabupaten Banyuwangi
No
Kecamatan
Rata-rata Koefisien Lokalisasi Tahun 2013-2017
Luas Lahan
Produksi
Penerimaan
1
Pesanggaran
0,0162
-0,0296
-0,0106
2
Siliragung
0,0342
-0,0800
-0,0264
3
Bangorejo
0,0926
0,0921
0,1916
4
Purwoharjo
0,1030
0,0448
0,0484
5
Tegaldlimo
0,0909
0,0830
0,1079
6
Muncar
0,0162
-0,0203
0,0013
7
Cluring
0,0616
0,0385
0,0595
8
Gambiran
0,0288
0,0185
0,0274
9
Tegalsari
0,0401
0,0051
0,0096
Sumber: Data Sekunder Diolah (2018). rkan Indikator Luas Lahan, Produksi dan Penerimaa Sumber: Data Sekunder Diolah (2018). kecamatan
yang
bersangkutan
memiliki
karakteristik iklim, cuaca, dan topografi yang
hampir sama sehingga Jeruk Siam dapat
dikembangkan dengan baik di kecamatan-
kecamatan
tersebut. Tersebarnya
kegiatan
usaha pertanian Jeruk Siam dapat pula
memberikan keuntungan bagi pelaku usaha-
usaha yang berkaitan dengan komoditas Jeruk
Siam karena apabila kebutuhan terhadap Jeruk
Siam tidak bisa diperoleh di suatu kecamatan
maka masih ada kecamatan-kecamatan lain
yang dapat dijadikan rujukan untuk memenuhi
kebutuhan akan Jeruk Siam. Hal ini sesuai
dengan
penelitian
yang
telah
dilakukan
Syamsiyah
et
al. Karakteristik Penyebaran Berdasarkan analisis lokalisasi indikator luas
lahan, produksi, dan penerimaan yang telah
dilakukan, dperoleh hasil yang tersaji terlihat
pada Tabel 6. p
Hasil analisis lokalisasi berdasarkan
indikator luas lahan, produksi dan penerimaan
pada Tabel 5. menunjukkan bahwa dari tahun
2013 sampai 2017 kecamatan-kecamatan yang
menjadi wilayah basis komoditas Jeruk Siam
berdasarkan
tiga
indikator
tersebut
menghasilkan angka koefisien lokalisasi yang
berfluktuatif. Hasil analisis lokalisasi juga
menunjukkan
bahwa
rata-rata
koefisien
lokalisasi pada kecamatan basis komoditas
Jeruk Siam bernilai positif kurang dari satu (α
(+)< 1). Hal tersebut berarti bahwa komoditas
Jeruk Siam tidak terlokalisasi pada suatu
kecamatan tertentu melainkan terlokalisasi
kecamatan yang menjadi kecamatan basis
komoditas Jeruk Siam. Tersebarnya kegiatan
usaha pertanian Jeruk Siam di Kabupaten
Banyuwangi
dikarenakan
kecamatan- Berdasarkan
analisis
spesialisasi
indikator luas lahan, produksi, dan penerimaan
yang telah dilakukan, diperoleh hasil yang
pada Tabel 7. Hasil
analisis
spesialisasi
bahwa
kecamatan basis komoditas Jeruk Siam yang
menghasilkan angka koefisien lokalisasi positif
kurang dari satu (α (+)<1) juga menghasilkan
angka koefisien spesialisasi positif kurang dari
satu (β(+)<1). Artinya bahwa sejak tahun 2013 93 Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Tabel 7. Hasil Analisis Spesialisasi Jeruk Siam Berdasarkan Indikator Luas Lahan, Produksi dan Penerimaan
Tahun 2013-2017 di Kabupaten Banyuwangi
No
Kecamatan
Rata-rata Koefisien Spesialisasi Tahun 2013-2017
Luas Lahan
Produksi
Penerimaan
1
Pesanggaran
0,0220
-0,4115
-0,3057
2
Siliragung
0,1253
-0,2592
-0,2189
3
Bangorejo
0,1618
0,1756
0,2207
4
Purwoharjo
0,4436
0,1525
0,1839
5
Tegaldlimo
0,2902
0,1402
0,2513
6
Muncar
0,0503
-0,3263
-0,1959
7
Cluring
0,2886
0,0238
0,1406
8
Gambiran
0,1838
0,0578
0,1471
9
Tegalsari
0,1606
-0,1797
-0,0125
Sumber: Data Sekunder Diolah (2018) Analisis Spesialisasi Jeruk Siam Berdasarkan Indikator Luas Lahan, Produksi dan Penerimaan
2013-2017 di Kabupaten Banyuwangi Sumber: Data Sekunder Diolah (2018) sampai tahun 2017 kecamatan basis komoditas
Jeruk Siam tidak menspesialisasikan komoditas
Jeruk Siam di wilayahnya. Hal tersebut
dikarenakan sebagian besar masyarakat atau
petani yang melakukan usaha budidaya Jeruk
Siam tidak menjadikan Jeruk Siam sebagai
tanaman utama yang dibudidayakan di lahan
mereka dengan kata lain ada komoditas
pertanian tertentu lainnya yang dijadikan
sebagai komoditas utama di wilayah mereka. Hal ini sesuai dengan penelitian yang telah
dilakukan Syamsiyah, dkk (2016) berdasarkan
koefisien spesialisasi tidak dihasilkan nilai
yang lebih ataupun sama dengan satu artinya di
Kabupaten Cirebon tidak satu kecamatanpun
yang melakukan spesialisasi pada kegiatan
usahatani tertentu. Kontribusi Komoditas Jeruk Siam terhadap
Perekonomian Kabupaten Banyuwangi Kontribusi Komoditas Jeruk Siam terhadap
Perekonomian Kabupaten Banyuwangi Kontribusi Komoditas Jeruk Siam terhadap
Perekonomian Kabupaten Banyuwangi
Hasil perhitungan kontribusi komoditas Jeruk
Siam
terhadap
perekonomian
Kabupaten
Banyuwangi ditampilkan dalam Tabel 9. Hasil perhitungan kontribusi komoditas Jeruk
Siam
terhadap
perekonomian
Kabupaten
Banyuwangi ditampilkan dalam Tabel 9. Berdasarkan Tabel 9. diketahui bahwa
kontribusi
Jeruk
Siam
terhadap
PDRB
perekonomian Kabupaten Banyuwangi dalam
kurun waktu 5 tahun berada pada kriteria
rendah. Hal
tersebut
dikarenakan
angka
presentase kontribusi Jeruk Siam masih lebih
kecil dari rata-rata kontribusi komponen
penyusun PDRB perekonomian Kabupaten
Banyuwangi (0,015%). Kontribusi Jeruk Siam
terhadap PDRB perekonomian dari tahun 2013
sampai dengan 2017 berturut-turut adalah
0,0019; 0,0041; 0,0102; 0,0050 dan 0,0020. Rendahnya kontribusi Jeruk Siam disebabkan
sumbangan Jeruk Siam masih rendah kepada
PDRB
subsektor
hortikultura
Kabupaten
Banyuwangi. Hal ini disebabkan ada 48
komoditas hortikultura yang dibudidayakan di
Kabupaten
Banyuwangi,
sehingga
nilai
penerimaan dari Jeruk Siam memberikan
kontribusi yang rendah terhadap perekonomian
Kabupaten Banyuwangi. Rendahnya kontribusi
Jeruk Siam dimungkinkan pula karena harga
Jeruk Siam yang berfluktuatif dari tahun 2013
sampai 2017 bahkan menunjukkan penurunan Karakteristik Penyebaran Kontribusi Komoditas Jeruk Siam Terhadap Perekonomian Kabupaten Banyuwangi Tahun 2013-
2017 ,
Keterangan: *Rata-rata kontribusi komponen penyusun PDRB Perekonomian Banyuwangi adalah 0,01 pada tahun 2017 sebesar 1,151 yang berarti
bahwa setiap kenaikan satu-satuan luas lahan,
produksi dan penerimaan Jeruk Siam di
Kabupaten Banyuwangi akan meningkatkan
luas lahan, produksi dan penerimaan Jeruk
Siam di wilayah basis sebesar hasil yang telah
diperoleh tersebut. Berdasarkan uraian tersebut
dapat dikatakan bahwa usaha pertanian Jeruk
Siam di Kabupaten Banyuwangi potensial
untuk dilakukan pengembangan. pada tahun 2017 sebesar 1,151 yang berarti
bahwa setiap kenaikan satu-satuan luas lahan,
produksi dan penerimaan Jeruk Siam di
Kabupaten Banyuwangi akan meningkatkan
luas lahan, produksi dan penerimaan Jeruk
Siam di wilayah basis sebesar hasil yang telah
diperoleh tersebut. Berdasarkan uraian tersebut
dapat dikatakan bahwa usaha pertanian Jeruk
Siam di Kabupaten Banyuwangi potensial
untuk dilakukan pengembangan. di dua tahun terakhir dan harga paling rendah
berada pada tahun 2017 yaitu hanya sebesar Rp
5500., per kilogram sehingga memberikan
peneimaan yang rendah pula untuk Kabupaten
Banyuwangi. KESIMPULAN DAN SARAN Berdasarkan hasil-hasil analisis yang telah
diperoleh, maka simpulan yang dapat diambil
dari penelitian ini adalah Kecamatan penghasil
Jeruk
Siam
di
Kabupaten
Banyuwangi
berdasarkan rata-rata nilai LQ tahun 2013
sampai 2017 yang secara konsisten selalu
menjadi kecamatan basis dengan tiga indikator
berbeda yaitu luas lahan, produksi dan
penerimaan adalah Kecamatan Bangorejo,
Kecamatan
Purwoharjo,
Kecamatan
Tegaldlimo,
Kecamatan
Cluring,
dan
Kecamatan Gambiran, karakteristik penyebaran
komoditas
Jeruk
Siam
di
Kabupaten
Banyuwangi pada tahun 2013 sampai dengan
2017
tidak
terlokalisasi
dan
tidak
terspesialisasi, serta kontribusi komoditas Jeruk
Siam
terhadap
perekonomian
Kabupaten
Banyuwangi pada tahun 2013 sampai dengan
2017 adalah rendah. Karakteristik Penyebaran Selama kurun waktu tahun
2013 sampai dengan 2017 di Kabupaten
Banyuwangi tidak satupun kecamatan basis
komoditas Jeruk Siam yang menspesialisasikan
kegiatan pertaniannya pada komoditas Jeruk
Siam saja. Besaran angka dampak pengganda (multiplier
effect) berdasarkan analisis Regional Multiplier
(RM) kegiatan budidaya Jeruk Siam di
Kabupaten Banyuwangi berdasarkan indikator
luas lahan, produksi dan penerimaan tahun
2013 sampai 2017 tersaji dalam Tabel 8. Tabel 8. Nilai Regional Multiplier (RM) Komoditas
Jeruk Siam di Kabupaten Banyuwangi
Berdasarkan
Indikator
Luas
Lahan,
Produksi dan Penerimaan Tahun 2013-
2017
Tahun
Nilai RM
Luas
Lahan
Produksi
Penerimaan
2013
1,007
1,104
1,105
2014
1,083
1,086
1,071
2015
1,054
1,078
1,073
2016
1,006
1,063
1,104
2017
1,014
1,143
1,151
Sumber: Data Sekunder Diolah (2018). Tabel 8. Nilai Regional Multiplier (RM) Komoditas
Jeruk Siam di Kabupaten Banyuwangi
Berdasarkan
Indikator
Luas
Lahan,
Produksi dan Penerimaan Tahun 2013-
2017 j
Menurut Pasaribu dan Soetriono (2009)
untuk menganalisa perkembangan daerah yang
dekat hubungannya dengan penelaahan siklus
daerah digunakan analisis Regional Multiplier
(RM). Analisis pengganda sederhana seperti ini
menggunakan
telaah
dasar
perekonomian
daerah yang membedakan antara sektor basis
dengan sektor non basis. Analisis Regional
Multiplier (RM) menunjukkan besarnya angka
pengganda atau efek perambatan dari suatu
kegiatan budidaya komoditas Jeruk Siam pada
suatu wilayah terhadap kegiatan budidaya
komoditas Jeruk Siam di wilayah basis. Berdasarkan hasil analisis Regional
Multiplier
(RM)
diketahui
bahwa
RM
menunjukkan angka lebih dari satu (RM>1). Artinya usaha budidaya Jeruk Siam di wilayah
Kabupaten Banyuwangi memberikan dampak
pengganda terhadap usaha budidaya Jeruk Siam
di
wilayah
basis. Nilai
RM
tertinggi
berdasarkan indikator luas lahan terdapat pada
tahun 2014 sebesar 1,083, nilai RM tertinggi
berdasarkan indikator produksi terdapat pada
tahun 2017 sebesar 1,143, nilai RM tertinggi
berdasarkan indikator penerimaan terdapat 94 Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Tabel 9. Kontribusi Komoditas Jeruk Siam Terhadap Perekonomian Kabupaten Banyuwangi Tahun 2013-
2017
Tahun
PDRB Jeruk Siam
PDRB Perekonomian
Kabupaten
Banyuwangi
Kontribusi
Jeruk Siam
Keterangan
(Miliar Rupiah)
(Miliar Rupiah)
(%)
2013
935,99
47.364.724,70
0,0019
Rendah
2014
2.186,15
53.371.199,30
0,0041
Rendah
2015
6.170,71
60.179.292,90
0,0102
Rendah
2016
3.331,77
66.348.468,12
0,0050
Rendah
2017
1.445,59
72.245.718,94
0,0020
Rendah
Sumber: Data sekunder diolah, 2018
Keterangan: *Rata-rata kontribusi komponen penyusun PDRB Perekonomian Banyuwangi adalah 0,015 % . DAFTAR PUSTAKA Boekoesoe, Y. 2011. Analisis Perwilayahan
Komoditas
Mangga
di
Provinsi
Gorontalo. Jurnal Ilmiah Agropolitan, 4
(2), 527-534. Djarwanto. 2001. Statistik Non Parametrik
Bagian 1 Edisi 3. Yogyakarta: BPFE-
UGM. 95 Rizqa N., Soetriono, Anik S.: Analisis Perwilayahan dan Kontribusi … Pemkab Banyuwangi. 2016. Rencana Strategis
Dinas Pertanian Kabupaten Banyuwangi
Tahun 2016-2021. Banyuwangi: Dinas
Pertanian Kabupaten Banyuwangi. Larasati,
N. 2011. Faktor-Faktor
yang
Mempengaruhi
Permintaan
dan
Penawaran Ubi Jalar serta Kontribusinya
terhadap Perekonomian Jawa Timur. Skripsi. Jember: Universitas Jember. Pemkab Banyuwangi. 2016. Rencana Strategis
Dinas Pertanian Kabupaten Banyuwangi
Tahun 2016-2021. Banyuwangi: Dinas
Pertanian Kabupaten Banyuwangi. Pusat Data dan Informasi Pertanian. 2015. Komoditas
Pertanian
Hortikultura:
Jeruk. Jakarta: Kementerian Pertanian. Nazir, M. 2005. Metode Penelitian. Jakarta:
Ghalia Indonesia. Mulyono, J. dan Munibah, K. 2016. Pendekatan
Location Quotient dan Shift Share
Analysis dalam Penentuan Komoditas
Unggulan
Tanaman
Pangan
di
Kabupaten
Bantul. Informatika
Pertanian, 25(2), 221-230. Soetriono. 1996. Sektor Basis Kedelai sebagai
Pendukung Agroindustri di Kecamatan
Ambulu Kabupaten Jember. Agrijurnal,
2(3), 1-13. Suharyanto, Agustian, A dan Silitonga, P.Y. 2013. Analisis Daya Saing Komoditas
Perkebunan Di Kabupaten Kaimana
Provinsi Papua Barat. SEPA, 10(1), 148-
155. Pasaribu,
A.P. dan
Soetriono. 2009. Perwilayahan
dan
Strategi
Pengembangan Komoditas Karet (Hevea
brasillensis) di Indonesia. J-SEP, 3(3), 1-
14. 96
|
https://openalex.org/W2593459708
|
https://www.nature.com/articles/srep43538.pdf
|
English
| null |
Circulating cathepsin-S levels correlate with GFR decline and sTNFR1 and sTNFR2 levels in mice and humans
|
Scientific reports
| 2,017
|
cc-by
| 9,107
|
Circulating cathepsin-S levels
correlate with GFR decline and
sTNFR1 and sTNFR2 levels in mice
and humans
September 2016
25 January 2017
OPEN received: 12 September 2016
accepted: 25 January 2017
Published: 27 February 2017 received: 12 September 2016
accepted: 25 January 2017
Published: 27 February 2017 Dominik Steubl1,*, Santhosh V. Kumar2,*, Maia Tato2, Shrikant R. Mulay2, Anders Larsson3,
Lars Lind3, Ulf Risérus4, Lutz Renders1, Uwe Heemann1, Axel C. Carlsson3,5, Johan Ärnlöv3,6 &
Hans-Joachim Anders2 Cardiovascular complications determine morbidity/mortality in chronic kidney disease (CKD). We
hypothesized that progressive CKD drives the release of cathepsin-S (Cat-S), a cysteine protease that
promotes endothelial dysfunction and cardiovascular complications. Therefore, Cat-S, soluble tumor-
necrosis-factor receptor (sTNFR) 1/2 and glomerular filtration rate (GFR) were measured in a CKD
mouse model, a German CKD-cohort (MCKD, n = 421) and two Swedish community-based cohorts
(ULSAM, n = 764 and PIVUS, n = 804). Association between Cat-S and sTNFR1/2/GFR was assessed
using multivariable linear regression. In the mouse model, Cat-S and sTNFR1/2 concentrations were
increased following the progressive decline of GFR, showing a strong correlation between Cat-S and
GFR (r = −0.746, p < 0.001) and Cat-S and sTNFR1/sTNFR2 (r = 0.837/0.916, p < 0.001, respectively). In the human cohorts, an increase of one standard deviation of estimated GFR was associated with a
decrease of 1.008 ng/ml (95%-confidence interval (95%-CI) −1.576–(−0.439), p < 0.001) in Cat-S levels
in MCKD; in ULSAM and PIVUS, results were similar. In all three cohorts, Cat-S and sTNFR1/sTNFR2
levels were associated in multivariable linear regression (p < 0.001). In conclusion, as GFR declines Cat-S
and markers of inflammation-related endothelial dysfunction increase. The present data indicating
that Cat-S activity increases with CKD progression suggest that Cat-S might be a therapeutic target to
prevent cardiovascular complications in CKD. Cathepsin-S (Cat-S) is a cysteine protease with intracellular and extracellular proteolytic activities1. Within endo-
somes, Cat-S is involved in the processing of the major histocompatibility class II complex during antigen pres-
entation, an essential element of host defense and autoimmunity2,3. In inflammatory states, Cat-S is released by
activated monocytes and neutrophils into the circulation where it contributes globally to endothelial dysfunction
by activating protease-activated receptor (PAR)-2 on microvascular endothelial cells to increase microvascu-
lar permeability and leukocyte adhesion4,5. Furthermore, release of Cat-S by activated macrophages within the
vascular wall degrades the elastic fibers of the tunica elastic, a process that contributes to vascular wall degen-
eration, media calcification, and aneurysm formation6–12. www.nature.com/scientificreports www.nature.com/scientificreports www.nature.com/scientificreports received: 12 September 2016
accepted: 25 January 2017
Published: 27 February 2017 Materials and Methods Animal studies. All experiments were performed in accordance with relevant guidelines and regulations and
were approved by the regional Government of Oberbayern, Germany. Male C57BL/6 mice were obtained from
Charles River (Sulzfeld, Germany) at the age of 7–8 weeks, mice were housed in groups of five in standard hous-
ing conditions with unlimited access to food and water for one week for acclimatization. CKD was induced by
feeding an oxalate-rich diet, which was prepared by adding 50 μmol/g sodium oxalate to a calcium-free standard
diet (Ssniff, Soest, Germany) as previously described22. Oxalate-rich diet was fed for 21 days. This model simulates
progressive CKD and its complications such as metabolic acidosis. Plasma samples were collected prior to diet
change and at days 1, 7, 14, and 21. All experimental procedures were approved by the local government authori-
ties. All plasma samples were analyzed for the levels of Cat-S, sTNFR1, and sTNFR2 using commercially available
enzyme-linked immunosorbent assay (ELISA) kits (Cloud-clone Corp. TX, USA, SEB933Mu; Sino Biologicals
Inc. PA, SEK50496 and SEK50128, respectively). Glomerular filtration rate measurement in conscious mice. Glomerular filtration rate (GFR)
was measured at different time points (day 0, 1, 7, 14 and 21) using an automated miniaturized imager device
(Medibeacon, Germany). The mouse neck was shaved and the device was mounted on the shaved area using
double-sided adhesive tape, then fluorescein-isothiocyanate(FITC)-sinistrin (Roche, Mannheim, Germany) was
injected intravenously to each mouse at the dose of 150 mg/kg. The transcutaneous FITC signal was recorded
continuously for a period of 1.5 h. Imaging data were analyzed using MPD Lab software (Medibeacon) and GFR
was calculated from the decrease of fluorescence intensity over time using a two-compartment calculation algo-
rithm as described23. Human cohorts and definition of chronic kidney disease stages. The Munich CKD cohort (MCKD)
consisted of 421 patients from the outpatient clinic at Klinikum rechts der Isar, Munich, Germany. Among them
were 69 patients without kidney disease (CKD 0°) serving as controls and 352 patients normally distributed
across stages I°-V° 24 of CKD. All participants provided written informed consent. The study adhered to the dec-
laration of Helsinki and was approved by the local ethics committee.h The Uppsala Longitudinal Study of Adult Men (ULSAM) was initiated in 1970. Materials and Methods All 50-year-old men born
between 1920 and 1924 and living in Uppsala, Sweden, were invited to participate in a health survey aimed
at identifying cardiovascular risk factors (13, described in detail at http://www.pubcare.uu.se/ULSAM). The
analyses presented in this study are based on the fourth examination cycle of ULSAM, in which participants
were approximately 77 years old (1998–2001). Of 1398 invited men, 838 (60%) participated. Of these, 89 were
excluded due to missing data on Cat-S, GFR or co-variates, leaving 749 participants as the present study sample. All 70-year-old individuals living in Uppsala, Sweden, between 2001 and 2004 were eligible for the Prospective
Investigation of the Vasculature in Uppsala Seniors (PIVUS) study (13, described in detail at http://www.medsci. uu.se/pivus/pivus.htm). In the present study the second examination cycle of PIVUS was used (2006–2009), in
which participants were 75 years old. Of 964 invited participants, 827 participated (86%). Of these, 23 participants
were excluded due to missing data on Cat-S, GFR or co-variates, leaving 804 participants as the present study
sample. All participants gave written informed consent and the ethics committee of Uppsala University approved
both study protocols. The studies adhere to the declaration of Helsinki. h
We applied the criteria for CKD stages according to the current Kidney Disease: Improving Global Outcomes
(KDIGO) guidelines24: “CKD is defined as abnormalities of kidney structure or function, present for >3 months,
with implications for health.” Therefore, the diagnosis of CKD was made when either estimated glomerular fil-
tration rate (eGFR) was <60 ml/min/1.73 m2 of body surface area (BSA) and/or apparent signs of kidney dam-
age were present. For diagnosis of kidney damage, we defined proteinuria with a cut-off >150 mg/g creatinine
(MCKD) or albuminuria >26.4 mg/g creatinine (ULSAM and PIVUS) on spot-urine specimen and/or histologi-
cally proven kidney disease and/or abnormalities detected in imaging techniques (ultrasound, computed tomog-
raphy, magnetic resonance imaging or nuclear imaging). Once the diagnosis of CKD was established, patients
were assigned to a certain stage of CKD according to the KDIGO recommendation: CKD I° when eGFR >90 ml/
min/1.73 m2 BSA, CKD II° 61–90 ml/min/1.73 m2 BSA, CKD 3° 31–60 ml/min/1.73 m2 BSA, CKD IV° 16–30 ml/
min/1.73 m2 BSA and CKD V° 0–15 ml/min/1.73 m2 BSA. Patients with a single kidney (e.g. due to nephrec-
tomy) were assumed to have CKD despite an eGFR >60 ml/min/1.73 m2 BSA. Circulating cathepsin-S levels
correlate with GFR decline and
sTNFR1 and sTNFR2 levels in mice
and humans
September 2016
25 January 2017
OPEN Indeed, circulating Cat-S levels have been identified
as a biomarker for age-related vascular diseases, hypertension, obesity, diabetes, and overall mortality in large
population-based cohort studies13–18. Recent data from mouse models suggest that selective inhibition of Cat-S
can attenuate renal inflammation in lupus nephritis and reduce atherosclerosis in CKD2,19. Therefore Cat-S might
be a therapeutic target in proinflammatory states, such as arterio-/atherosclerosis. 1Abteilung für Nephrologie, Klinikum rechts der Isar, Technische Universität München, Munich, Germany. 2Medizinische Klinik und Poliklinik IV, Renal Division, Klinikum der Universität München, Campus Innenstadt,
München, Germany. 3Department of Medical Sciences, Uppsala University, Uppsala, Sweden. 4Departments of
Public Health and Caring Sciences/Clinical Nutrition, Uppsala University, Uppsala, Sweden. 5Division of Family
Medicine, Department of Neurobiology, Care Sciences and Society, Karolinska Institutet, Huddinge, Sweden. 6School of Health and Social Studies, Dalarna University, Falun, Sweden. *These authors contributed equally to this
work. Correspondence and requests for materials should be addressed to D.S. (email: dominik.steubl@lrz.tum.de) SCIeNTIFIC ReporTS | 7:43538 | DOI: 10.1038/srep43538 1 www.nature.com/scientificreports/ Chronic kidney disease (CKD) is a systemic inflammatory state that dramatically accelerates cardiovascular
morbidity and mortality20,21. The causal relationships between uremia, systemic inflammation and cardiovascular
disease in CKD are not entirely clear. Based on the findings mentioned above, we hypothesized that CKD and sys-
temic inflammation are associated with increasing levels of circulating Cat-S. To test this hypothesis, in a mouse
model of progressive CKD and in three clinical cohorts of CKD patients we quantified glomerular filtration rate
(GFR) as a measure of CKD, serum levels of soluble tumor necrosis factor-alpha receptors (sTNFR1 and sTNFR2)
as a measure of systemic inflammation, and serum/plasma concentrations of Cat-S. Materials and Methods Demographic characteristics and laboratory parameters in MCKD cohort. Data are presented as
mean ± standard deviation or as absolute numbers (n) with percentage (%) in brackets; CKD = chronic kidney
disease; BMI = body mass index; BUN = blood-urea nitrogen; eGFR calculated from CKD-EPIcreatinine-cystatin c
formula; BSA = body surface area; sTNFR = soluble tumor-necrosis-factor receptor; CRP = c-reactive protein;
protein/creatinine ratio in urine; antihypertensive medication also includes ACE-inhibitors/AT1-receptor
antagonists, which also might have been used for solely antihypertensive medication; therefore, the number of
patients with antihyperrtensive medication exceeds the number of patients with arterial hypertension in CKD
groups. latex-enhanced reagent (NLatexCystatin C; Siemens, Deerfield, IL, USA) detected using a BN ProSpec analyser
(Siemens) in ULSAM; or with latex-enhanced reagents (Gentian, Moss, Norway) detected using an Architect
ci8200 (Abbott Laboratories, Abbott Park, IL, USA) in PIVUS. In MCKD, the eGFR equation was calculated
based on serum creatinine and cystatin C concentrations, as published25. In ULSAM and PIVUS, eGFR was
assessed by a sole cystatin C-equations, as no data on circulating creatinine was available in these cohorts26. High-sensitive CRP measurements were performed using a latex-enhanced reagent (Siemens) assayed on a BN
ProSpec® analyzer (Siemens) in ULSAM; with the use of an Architect ci8200 in PIVUS; or using a turbidimetric
measurement in MCKD. Serum levels of Cat-S were measured by an ELISA (human cathepsin S [total] DY1183;
R&D Systems, Minneapolis, Minnesota). In the MCKD cohort, blood samples were frozen for a median of 515
days (range 340–677 days) until Cat-S levels were quantified in November 2015. In the ULSAM cohort, blood
samples were frozen for a median of 10.0 years (range 8.7–12.4 years) and in the PIVUS study, for 2.0 years
(range, 0.3–4.0 years) until analysis of Cat-S. The intra-assay coefficient of variation was 7%. To date, there are
no established reference values for circulating Cat-S in humans. Soluble TNFR-1 and TNFR-2 were measured as
previously described26. In ULSAM, we also analyzed urinary Cat-S levels in 513 individuals using the same assay
as above. Urine creatinine was analyzed with a modified kinetic Jaffe reaction on an Architect Ci8200® analyzer
(Abbott, Abbot Park, IL, USA) and Cat-S/creatinine ratio (in μg/mol) was calculated. Statistical Analysis. For statistical analysis IBM SPSS 20 (IBM, Armonk, USA) and Stata 12 (Stata Corp.,
USA) were used. Continuous data are expressed as mean with standard deviation (SD), categorical variables are
reported in absolute numbers and percentages. Materials and Methods 69 patients in MCKD without a
medical history for CKD and an eGFR >60 ml/min/1.73 m2 BSA were assigned to the group CKD 0°. Patients’
demographics of MCKD and PIVUS/ULSAM are given in Table 1, 2 and 3. Standard methods were used to assess
anthropometrical measurements, blood pressure, and blood sampling. Venous blood samples were drawn in the
morning after an overnight fast and stored at −80 °C. Biomarker analyses. For biomarker analysis, plasma was used in MCKD and serum in PIVUS/ULSAM. Creatinine and blood-urea-nitrogen (BUN) were measured using photometric techniques (creatinine: nor-
mal range 0.7–1.3 mg/dl in males and 0.5–1.1 mg/dl in females; BUN: normal range 7–18 mg/dl). Materials and Methods In MCKD,
cystatin C levels were assessed using a nephelometric immunoassay (normal range 0,50–0,96 mg/l); or using a SCIeNTIFIC ReporTS | 7:43538 | DOI: 10.1038/srep43538 2 www.nature.com/scientificreports/ MCKD
Total (n = 421)
CKD 0°
(n = 69, 16.4%)
CKD I°
(n = 37, 8.8%)
CKD II°
(n = 68, 16.2%)
CKD III°
(n = 126, 29.9%)
CKD IV°
(n = 80, 19.0%)
CKD V°
(n = 41, 9.7%)
Age (years, SD)
56.8 (±16.4)
49.1(±15.1)
41.6 (±14.9)
52.3 (±14.3)
63.8 (±13.6)
62.2 (±16.2)
59.1 (±14.9)
Gender (male, %)
246 (58.4)
29 (42.0)
19 (51.4)
39 (57.4)
80 (63.5)
52 (65.0)
27 (65.9)
BMI (kg/m2, SD)
26.9 (±5.7)
25.0 ± 6.0
26.1 (±5.6)
26.6 (±5.6)
26.9 (±5.0)
27.6 (±6.0)
28.5 (±6.1)
Diabetes mellitus (n, %)
96 (22.8)
5 (7.2)
3 (8.1)
8 (11.8)
30 (23.8)
29 (36.3)
21 (29.3)
Antidiabetic medication (n, %)
72 (17.1)
5 (7.2)
3 (8.1)
5 (7.4)
28 (22.2)
22 (27.5)
9 (22.0)
Arterial hypertension (n, %)
277 (65.8)
13 (18.8)
21 (56.8)
39 (57.4)
99 (78.6)
66 (82.5)
39 (95.1)
Systolic blood pressure (mmHg, SD)
137.5 (±20.1)
135.7 (±20.4)
137.1 ± 20.6
133.5 (±17.8)
136.1 (±19.9)
140.9 (±18.1)
144.0 (±25.6)
Diastolic blood pressure (mmHg, SD)
82.8 (±12.3)
83.1 (±10.9)
86.4 ± 11.8
82.7 (±11.7)
82.7 (±12.4)
81.1 (±13.5)
82.8 (±12.0)
Antihypertensive medication (n)
320
13
26
55
114
72
40
Cardiovascular disease (n, %)
56 (13.3)
2 (2.9)
2 (5.4)
4 (5.9)
22 (17.5)
18 (22.5)
8 (19.5)
Creatinine (mg/dl, SD)
2.0 (±1.7)
0.9 (±0.2)
1.0 (±0.3)
1.3 (±0.3)
1.7 (±0.4)
2.8 (±0.8)
6.0 (±2.6)
Cystatin C (mg/l, SD)
1.53 (±0.96)
0.65 (±0.17)
0.58 (±0.18)
0.91 (0.16)
1.44 (±0.29)
2.41 (±0.50)
3.52 ±0.63)
BUN (mg/dl, SD)
29.8 (±17.9)
13.5 (±3.8)
14.8 (±4.8)
19.3 (±5.2)
27.3 (±9.2)
43.1 (±12.7)
63.9 (±16.8)
eGFR (ml/min/1.73 m2 BSA, SD)
57.9 (±37.2)
108.2 (±21.8)
113.1 (±20.5)
73.1 (±8.7)
43.6 (±8.4)
22.6 (±4.6)
11.5 (±3.0)
Cathepsin-S, (ng/ml, SD)
12.5 (±5.0)
10.8 (±4.1)
11.2 (±8.5)
11.6 (±4.1)
12.9 (±4.4)
14.0 (±5.3)
13.5 (±3.8)
sTNFR1 (ng/ml, SD)
1.94 (±2.07)
0.67 (±0.62)
0.52 (±0.59)
0.83 (±0.57)
1.51 (±0.93)
3.44 (±1.74)
5.59 (±3.03)
sTNFR2 (ng/ml, SD)
11.6 (±16.7)
7.1 (±23.3)
4.5 (±2.8)
5.3 (±3.1)
10.3 (±13.8)
20.2 (±21.0)
23.4 (±7.9)
CRP (mg/dl, SD)
0.7 (±1.6)
0.3 (±0.4)
0.2 (±0.3)
0.5 (±1.0)
0.7 (±1.4)
1.1 (±2.1)
1.4 (±2.8)
Protein/creatinine ratio (mg/g, SD)
872.6 (±1697.9)
59.6 (±63.1)
827.3 (±1523.1)
474.2 (±809.7)
643.3 (±1164.7)
1875.4 (±2791.4)
2143.3 (±1609.9)
Underlying disease
None (n, %)
69 (16.4)
69 (100)
—
—
—
—
—
Diabetic nephropathy (n, %)
26 (6.2)
—
0 (0)
1 (1.5)
10 (7.9)
9 (11.3)
6 (14.6)
Hypertensive nephropathy (n, %)
58 (13.8)
—
4 (10.8)
7 (10.3)
16 (12.7)
19 (23.8)
12 (29.3)
Glomerulonephritis (n, %)
112 (26.6)
—
17 (45.9)
26 (38.2)
39 (31.0)
22 (27.5)
8 (19.5)
Polycystic kidney disease (n, %)
15 (3.6)
—
4 (10.8)
3 (4.4)
4 (3.2)
2 (2.5)
2 (4.9)
Postrenal failure (n, %)
29 (6.9)
—
1 (2.7)
9 (13.2)
13 (10.3)
4 (5.0)
2 (4.9)
Others/unknown (n, %)
112 (26.6)
—
11 (29.7)
22 (32.4)
44 (34.9)
24 (30.0)
11 (26.8) Table 1. Table 1. Demographic characteristics and laboratory parameters in MCKD cohort. Data are presented as
mean ± standard deviation or as absolute numbers (n) with percentage (%) in brackets; CKD = chronic kidney
disease; BMI = body mass index; BUN = blood-urea nitrogen; eGFR calculated from CKD-EPIcreatinine-cystatin c
formula; BSA = body surface area; sTNFR = soluble tumor-necrosis-factor receptor; CRP = c-reactive protein;
protein/creatinine ratio in urine; antihypertensive medication also includes ACE-inhibitors/AT1-receptor
antagonists, which also might have been used for solely antihypertensive medication; therefore, the number of
patients with antihyperrtensive medication exceeds the number of patients with arterial hypertension in CKD
groups. Cathepsin-S serum levels increase following the decline of glomerular filtration rate in mice. Cathepsin-S serum levels increase following the decline of glomerular filtration rate in mice. To address our hypothesis we first studied serum levels of Cat-S in a mouse model of progressive CKD22. Feeding
mice a sodium oxalate-rich and calcium-depleted diet resulted in a progressive nephrocalcinosis and a con-
tinuous decline of GFR, similar to CKD in children with primary hyperoxaluria type 127. Serum samples were
obtained at several time points and Cat-S levels were found to increase as CKD progressed with a correlation
coefficient of -0.7459 between Cat-S and GFR (p < 0.001, Fig. 1A). Next, we measured serum levels of sTNFR1
and sTNFR2, two established biomarkers of inflammation-related endothelial dysfunction, known to correlate
with CKD progression and to predict cardiovascular mortality in CKD26. Both sTNFRs showed the same negative
correlation with GFR decline (p < 0.001 respectively, Fig. 1B and C). Furthermore the pathophysiologic conse-
quences of oxalate diet-induced CKD could be observed in the mouse kidney morphology using silver staining
(Fig. 1D). The histological examination confirmed a progressive tubular atrophy and fibrosis at different time
intervals after feeding the oxalate diet, as well as an accumulation of F4/80 positive macrophages during the time
course of CKD progression. Cathepsin-S serum levels are negatively correlated with estimated glomerular filtration rate
in German CKD patients. In patients of the Munich CKD cohort (MCKD), plasma Cat-S concentrations
progressively increased from CKD I° to CKD V° (Table 1, Fig. 2A). Compared to CKD 0°, the groups CKD
III° (p = 0.002), IV° and V° (p < 0.001 respectively) showed significantly higher Cat-S levels, whereas CKD I°
(p = 0.674) and CKD II° (p = 0.278) did not differ significantly from CKD 0°. sTNFR1 and sTNFR2 concentra-
tions also increased from CKD I° to CKD V° (Table 1). In univariate analysis including both non-CKD and CKD
patients, Cat-S was inversely correlated to eGFR (rho (r) = −0.244, p < 0.001, Fig. 2A) and was positively associ-
ated with creatinine (r = 0.167, p = 0.001), BUN (r = 0.166, p = 0.001), cystatin C (r = 0.231, p < 0.001) as well as
both sTNFR1 (r = 0.304, p < 0.001, Fig. 2B) and sTNFR2 (r = 0.178, p < 0.001, Fig. 2C). Materials and Methods Normality of distribution for all parameters tested was tested
using Shapiro-Wilk-test. Differences of biomarker concentrations between patient groups were evaluated using
student’s t-test. Univariate correlations between Cat-S and eGFR, sTNFR1/2 were evaluated using Spearman-rank
coefficient in the mouse model and Pearson-correlation coefficient in the human cohorts. Additionally in MCKD,
correlations between Cat-S and blood creatinine, BUN and cystatin C were assessed using Pearson-coefficient. Materials and Methods Data
are presented as mean ± standard deviation or as absolute numbers (n) with percentage (%) in brackets;
CKD = chronic kidney disease; BMI = body mass index; LDL = low-density lipoprotein; HDL = high-density
lipoprotein; eGFR calculated from cystatin c formula; BSA = body surface area; sTNFR = soluble tumor-
necrosis-factor receptor; CRP = c-reactive protein; Albumin/creatinine ratio in urine. To adjust the association between Cat-S (dependent variable) and eGFR/sTNFRs (independent variables) for
demographic (age, gender, body-mass-index) and inflammation parameters (CRP) multivariable linear regres-
sion modelling was applied in the human cohorts. To assess the association between Cat-S and eGFR, the latter
was transformed into a standardized scale. Since sTNFRs (independent variables) were not sufficiently normally
distributed, both were transformed into logarithmic scales in the referring linear regression model. In secondary
analyses we also investigated the association between urinary Cat-S/creatinine ratio with eGFR using univariate
Spearman-Rank correlation coefficient. All reported p-values are two-sided, with a significance level of 0.05. Statistical evaluation was performed similarly in all three cohorts included in the study. To adjust the association between Cat-S (dependent variable) and eGFR/sTNFRs (independent variables) for
demographic (age, gender, body-mass-index) and inflammation parameters (CRP) multivariable linear regres-
sion modelling was applied in the human cohorts. To assess the association between Cat-S and eGFR, the latter
was transformed into a standardized scale. Since sTNFRs (independent variables) were not sufficiently normally
distributed, both were transformed into logarithmic scales in the referring linear regression model. In secondary
analyses we also investigated the association between urinary Cat-S/creatinine ratio with eGFR using univariate
Spearman-Rank correlation coefficient. All reported p-values are two-sided, with a significance level of 0.05. Statistical evaluation was performed similarly in all three cohorts included in the study. Materials and Methods SCIeNTIFIC ReporTS | 7:43538 | DOI: 10.1038/srep43538 3 www.nature.com/scientificreports/ ULSAM
Total
(n = 749)
CKD 0°
(n = 85, 11.4%)
CKD I°
(n = 20, 2.7%)
CKD II°
(n = 491, 65.6%)
CKD III°
(n = 142, 19.0%)
CKD IV°
(n = 11, 1.5%)
CKD V°
(n = 0)
Age (years, SD)
77.6 (±0.78)
77.4 (±0.83)
77.6 (±0.79)
77.6 (±0.78)
77.6 (±0.68)
77.8 (±0.74)
—
BMI (kg/m2, SD)
26.3 (±3.5)
25.9 ± 2.8
26.1 (±5.6)
26.2 (±3.39)
27.0 (±4.0)
26.8 (±4.3)
—
Diabetes mellitus (n, %)
82 (10.9)
13 (15.3)
3 (15.0)
1 (0.2)
61 (43.0)
4 (3)
—
Antidiabetic medication (n, %)
66 (8.8)
7 (8.2)
1 (5.0)
37 (7.5)
19 (13.4)
2 (18.2)
—
Arterial hypertension (n, %)
332 (44.3)
28 (32.9)
7 (35.0)
193 (39.3)
96 (67.6)
8 (72.3)
—
Systolic blood pressure (mmHg, SD)
150 (±21)
147 (±21)
149 ± 20
151 (±20)
150 (±21.4)
161 (±31)
—
Diastolic blood pressure (mmHg, SD)
81.3 (±9.8)
79.3 (±9.1)
81.0 ± 13.3
81.7 (±9.4)
81.1 (±10.8)
80.7 (±7.7)
—
Antihypertensive medication (n, %)
307 (41.0)
27 (31.8)
6 (30.0)
174 (35.4)
92 (64.8)
8 (72.7)
—
Cardiovascular disease (n, %)
204 (27.2)
17 (20.0)
1 (5.0)
118 (24.0)
62 (43.7)
6 (54.5)
—
Lipid-lowering therapy (n, %)
127 (17.0)
17 (20.0)
2 (10.0)
72 (14.7)
33 (23.2)
3 (27.3)
—
LDL (mmol/l, SD)
3.5 (±0.9)
3.6 (±0.9)
3.7 (±0.7)
3.5 (±0.9)
3.4 (±0.9)
3.3 (±1.0)
—
HDL (mmol/l, SD)
1.3 (±0.3)
1.4 (±0.4)
1.5 (±0.3)
1.3 (±0.3)
1.2 (±0.3)
1.1 (±0.2)
—
Cystatin C (mg/l, SD)
1.09 (±0.28)
0.82 (±0.05)
0.83 (±0.04)
1.03 (±0.09)
1.41 (±0.17)
2.58 (±0.28)
—
eGFR (ml/min/1.73 m2 BSA, SD)
73.3 (±17.1)
100.2 (±9.1)
98.0 (±6.9)
75.1 (±8.1)
51.2 (±7.0)
23.9 (±3.5)
—
Cathepsin-S, (ng/ml, SD)
14.3 (±4.0)
13.4 (±3.5)
15.1 (±3.2)
14.1 (±3.9)
15.1 (±4.4)
17.4 (±5.9)
—
sTNFR1 (ng/ml, SD)
2.07 (±0.81)
1.61 (±0.49)
1.16 (±0.60)
2.0 (±0.64)
1.51 (±0.80)
5.06 (±1.9)
—
CRP (mg/dl)
0.4 (±0.7)
0.3 (±0.5)
0.4 (±0.7)
0.4 (±0.5)
0.6 (±1.2)
0.5 (±0.2)
—
Albumin/creatinine ratio (mg/g, SD)
38.9 (±169.00)
6.8 (±5.5)
50.0 (±36.6)
26.4 (±93.3)
45.2 (±100.3)
836.0 (±1001.4)
— Table 2. Demographic characteristics and laboratory parameters in ULSAM cohort (100% male). Cathepsin-S serum levels increase following the decline of glomerular filtration rate in mice. In multivariable linear
regression analysis, an increase of one standard deviation in eGFR (37.2 ml/min/1.73 m2 BSA) was associated
with a decrease of 1.008 ng/ml in Cat-S concentration (p < 0.001, Table 4). Cat-S was also positively associated
with (log) sTNFR1 and (log) sTNFR2 in multivariable regression analysis: one unit increase of (log) TNFR1 was
associated with an increase of 1.314 ng/ml of Cat-S (p < 0.001, Table 5); an increase of one unit of (log) TNFR2
was associated with an increase of 1.976 ng/ml of Cat-S (p < 0.001, Table 5). Cathepsin-S serum levels increase following the decline of glomerular filtration rate in mice. SCIeNTIFIC ReporTS | 7:43538 | DOI: 10.1038/srep43538 4 www.nature.com/scientificreports/ PIVUS
Total
(n = 804)
CKD 0°
(n = 69, 8.6%)
CKD I°
(n = 18, 2.2%)
CKD II°
(n = 443, 55%)
CKD III°
(n = 261, 32%)
CKD IV°
(n = 13, 1.6%)
CKD V°
(n = 0, 0%)
Age (years, SD)
75.3 (±0.19)
75.3 (±0.18)
75.3 (±0.19)
75.3 (±0.18)
75.3 (±0.19)
75.3 (±0.23)
—
Gender (male, %)
408 (50.8)
47 (68.1)
7 (38.9)
245 (55.3)
105 (40.2)
4 (30.7)
—
BMI (kg/m2, SD)
26.8 (±4.3)
25.4 ± 3.5
23.6 (±3.0)
26.5 (±4.1)
28.0 (±4.6)
27.7 (±5.8)
—
Diabetes mellitus (n, %)
93 (11.6)
7 (10.1)
1 (5.5)
41 (9.3)
40 (15.3)
4 (30.7)
—
Antidiabetic medication (n, %)
72 (9.0)
4 (5.8)
1 (5.6)
32 (7.2)
32 (12.3)
3 (23.1)
—
Arterial hypertension (n, %)
654 (81.3)
47 (68.2)
16 (88.9)
360 (81.3)
218 (83.5)
13 (100)
—
Systolic blood pressure (mmHg, SD)
149 (±19)
147 (±18.6)
155 (± 21.2)
148 (±18.6)
149 (±20.7)
150 (±21.9)
—
Diastolic blood pressure (mmHg, SD)
75.9 (±9.4)
73.5 (±10.2)
80.2 ± 9.8
76.2 (±9.0)
75.9 (±9.5)
72.8 (±14.4)
—
Antihypertensive medication (n)
389 (48.4)
22 (31.9)
7 (38.9)
187 (42.2)
162 (62.1)
11 (84.6)
—
Cardiovascular disease (n, %)
68 (8.5)
3 (4.3)
2 (11.1)
18 (4.1)
40 (15.3)
5 (38.5)
—
Lipid-lowering therapy (n, %)
206 (25.6)
19 (27.5)
4 (22.2)
104 (23.5)
74 (28.4)
5 (38.5)
—
LDL (mmol/l, SD)
3.4 (±0.9)
3.5 (±0.9)
3.6 (±1.4)
3.5 (±0.9)
3.1 (±0.9)
3.2 (±0.8)
—
HDL (mmol/l, SD)
1.5 (±0.5)
1.8 (±0.5)
1.5 (±0.4)
1.5 (±0.4)
1.3 (±0.4)
1.3 (±0.5)
—
Cystatin C (mg/l, SD)
0.92 (±0.18)
0.73 (±0.08)
0.74 (±0.09)
0.87 (±0.09)
1.05 (±0.16)
1.52 (±0.40)
—
eGFR (ml/min/1.73 m2 BSA, SD)
67.8 (±19.3)
102.2 (±20.3)
102.6 (±10.7)
72.9 (±8.0)
50.1 (±7.2)
21.0 (±7.5)
—
Cathepsin-S, (ng/ml, SD)
20.4 (±7.4)
17.9 (±5.6)
18.6 (±6.7)
20.3 (±7.1)
21.1 (±7.9)
25.0 (±11.2)
—
sTNFR1 (ng/ml, SD)
2.46 (±1.3)
1.81 (±0.35)
1.82 (±0.37)
2.18 (±0.44)
2.93 (±1.43)
3.44 (±1.74)
—
sTNFR2 (ng/ml, SD)
6.33 (±2.89)
4.68 (±1.41)
6.21 (±7.34)
5.71 (±1.66)
7.42 (±2.49)
6.76 (±5.49)
—
CRP (mg/dl)
0.2 (±0.3)
0.2 (±0.3)
0.2 (±0.3)
0.2 (±0.4)
0.2 (±0.3)
0.2 (±0.1)
—
Albumin/creatinine ratio (mg/g, SD)
54.6 (±255.2)
11.3 (±6.0)
111.8 (±115.3)
33.1 (±139.0)
90.2 (±398.6)
294.8 (±356.4)
—
Table 3. Demographic characteristics and laboratory parameters in PIVUS cohort. Cathepsin-S serum levels increase following the decline of glomerular filtration rate in mice. Data are presented as
mean ± standard deviation or as absolute numbers (n) with percentage (%) in brackets; CKD = chronic kidney
disease; BMI = body mass index; LDL = low-density lipoprotein; HDL = high-density lipoprotein; eGFR
calculated from cystatin c formula; BSA = body surface area; sTNFR = soluble tumor-necrosis-factor receptor;
CRP = c-reactive protein; Table 3. Demographic characteristics and laboratory parameters in PIVUS cohort. Data are presented as
mean ± standard deviation or as absolute numbers (n) with percentage (%) in brackets; CKD = chronic kidney
disease; BMI = body mass index; LDL = low-density lipoprotein; HDL = high-density lipoprotein; eGFR
calculated from cystatin c formula; BSA = body surface area; sTNFR = soluble tumor-necrosis-factor receptor;
CRP = c-reactive protein; Cathepsin-S serum levels are negatively correlated with glomerular filtration rate in two
Swedish population cohorts. To validate the data from the MCKD cohort using unrelated and unselected
population cohorts, we analyzed two population-based samples, ULSAM and PIVUS from the Uppsala region
in Sweden. Similarly to MCKD, Cat-S concentrations gradually increased with deterioration of kidney function
in both cohorts (Tables 2 and 3). In univariate analysis, serum Cat-S was negatively correlated with eGFR in
both cohorts (p < 0.001 respectively, Figs 3A and 4A). Analogous to MCKD, Cat-S was positively associated
with sTNFR1 in both ULSAM (p < 0.001, Fig. 3B) and PIVUS (p < 0.001, Fig. 4B). sTNFR2 was only available
in PIVUS, in which it also positively associated with Cat-S (p < 0.001, Fig. 4C). In multivariable linear regres-
sion analysis, an increase of one standard deviation of eGFR (±17.1 ml/min/1.73 m2 BSA in ULSAM, ±19.3 ml/
min/1.73 m2 BSA in PIVUS) resulted in a 0.664 ng/ml decrease of Cat-S serum concentrations (p < 0.001, Table 4)
in ULSAM and a 1.076 ng/ml decrease (p < 0.001, Table 4) in PIVUS. Cat-S was also significantly associated
with (log) sTNFR1 in both ULSAM and PIVUS: a one-unit increase of (log) sTNF1 was associated with a Cat-S
increase of 2.474 ng/ml in ULSAM and 4.034 ng/ml in PIVUS (p < 0.001 respectively, Table 5). In PIVUS, Cat-S
and (log) sTNFR2 showed a significant association, with a Cat-S increase of 8.935 ng/ml per unit increase of
(log) sTNF2 (p < 0.001, Table 5). There was no association between urinary Cat-S/creatinine ratio with eGFR
(p = 0.465, Supplementary Figure 1). Cathepsin-S serum levels increase following the decline of glomerular filtration rate in mice. (p
,
pp
y
g
)
Taken together, we conclude that Cat-S serum levels show an negative correlation with renal function as
defined by measured (mice) or estimated (human) GFR. This inverse correlation is identical to those of sTNFR1
and sTNFR2, two validated markers of systemic inflammation-related endothelial dysfunction. Discussion We hypothesized that the progression of CKD and its related systemic inflammatory state is associated with
increasing levels of circulating Cat-S, a protease involved in the pathogenesis of cardiovascular disease and a
potential therapeutic target to attenuate atherosclerosis in CKD19. Our approach to address this issue included
experiments in which we induced CKD in mice, and cross-sectional analyses of three European human cohorts,
one of selected CKD patients (MCKD) and two population-based cohorts (PIVUS, ULSAM) of elderly individ-
uals. Our data confirm the hypothesis and reveal a consistent negative correlation between Cat-S serum levels
and measured GFR (mice) or estimated GFR (humans). In addition, we found a positive correlation of Cat-S
levels with markers of glomerular function (blood creatinine, BUN and cystatin C) as well as with two established
markers of systemic inflammation in CKD patients, sTNFR1 and sTNFR226. This shows a robust association of
Cat-S levels with different markers of glomerular function and inflammation.h f
gl
These results establish the impact of renal function on Cat-S concentrations in the circulation. We recently
reported that Cat-S is filtered across the glomerular filtration barrier5. Because proximal tubular cells of mouse
and human kidneys contain Cat-S protein but no RNA transcripts, intrarenal Cat-S mRNA (and protein) expres-
sion largely originates from infiltrating CD68+ macrophages that (unlike tubular epithelial cells) do not express SCIeNTIFIC ReporTS | 7:43538 | DOI: 10.1038/srep43538 5 www.nature.com/scientificreports/ ntificreports/
Figure 1. Mouse model of progressive chronic kidney disease: Plasma samples were collected at different
time points in oxalate-rich diet fed mice as per the different stages of CKD. We analyzed serum levels of
Cat-S (A), sTNFR1 (B), and sTNFR2 (C) plotted with GFR at day 0, 1, 7, 14 and 21. D: Representative images
of silver staining and F4/80 positive macrophages (source of Cat-S) in the kidneys of oxalate-rich diet fed mice
at different time points. Data expressed as absolute individual values from 5–6 mice each group and Spearman
correlation analysis was performed against change in GFR; level of significance for the correlations between
GFR and Cat-S, sTNFR1 and sTNFR2 was p < 0.001, respectively. Figure 1. Mouse model of progressive chronic kidney d
time points in oxalate-rich diet fed mice as per the differ
Cat-S (A), sTNFR1 (B), and sTNFR2 (C) plotted with GFR
of silver staining and F4/80 positive macrophages (source
at different time points. Discussion Data expressed as absolute individ
correlation analysis was performed against change in GFR
GFR and Cat-S, sTNFR1 and sTNFR2 was p < 0.001, respe
Study
group
regression
coefficient B
95% confidence
intervals
p-value Figure 1. Mouse model of progressive chronic kidney d
time points in oxalate-rich diet fed mice as per the differ
Cat-S (A), sTNFR1 (B), and sTNFR2 (C) plotted with GFR
of silver staining and F4/80 positive macrophages (source
at different time points. Data expressed as absolute individ
l ti
l
i
f
d
i
t h
i GFR Figure 1. Mouse model of progressive chronic kidney disease: Plasma samples were collected at different
time points in oxalate-rich diet fed mice as per the different stages of CKD. We analyzed serum levels of
Cat-S (A), sTNFR1 (B), and sTNFR2 (C) plotted with GFR at day 0, 1, 7, 14 and 21. D: Representative images
of silver staining and F4/80 positive macrophages (source of Cat-S) in the kidneys of oxalate-rich diet fed mice
at different time points. Data expressed as absolute individual values from 5–6 mice each group and Spearman
correlation analysis was performed against change in GFR; level of significance for the correlations between
GFR and Cat-S, sTNFR1 and sTNFR2 was p < 0.001, respectively. Study
group
regression
coefficient B
95% confidence
intervals
p-value
MCKD
−1.008
−1.576–(−0.439)
<0.001
ULSAM
−0.664
−0.952–(−0.375)
<0.001
PIVUS
−1.076
−1.604–(−0.547)
<0.001
Table 4. Multivariable linear regression models showing the association between standard deviation
increments in eGFR (independent variable, in ml/min/1.73 m2 body surface area (BSA)) and cathepsin-S
(dependent variable, in ng/ml) in all three study cohorts. Multivariable analysis adjusted for age, gender,
body-mass-index and c-reactive protein; the regression coefficient B describes the change of cathepsin-S
(in ng/ml) per change of one standard deviation (SD) of eGFR in every cohort (SD in MCKD 37.2 ml/
min/1.73 m2 BSA, in ULSAM 17.1 ml/min/1.73 m2 BSA, in PIVUS 19.3 ml/min/1.73 m2 BSA). Study
group
regression
coefficient B
95% confidence
intervals
p-value
MCKD
−1.008
−1.576–(−0.439)
<0.001
ULSAM
−0.664
−0.952–(−0.375)
<0.001
PIVUS
−1.076
−1.604–(−0.547)
<0.001
Table 4. Multivariable linear regression models showing the association between standard deviation
increments in eGFR (independent variable, in ml/min/1.73 m2 body surface area (BSA)) and cathepsin-S
(dependent variable, in ng/ml) in all three study cohorts. Discussion Multivariable analysis adjusted for age, gender,
body-mass-index and c-reactive protein; the regression coefficient B describes the change of cathepsin-S
(in ng/ml) per change of one standard deviation (SD) of eGFR in every cohort (SD in MCKD 37.2 ml/
min/1.73 m2 BSA, in ULSAM 17.1 ml/min/1.73 m2 BSA, in PIVUS 19.3 ml/min/1.73 m2 BSA). Study
group
regression
coefficient B
95% confidence
intervals
p-value
MCKD
−1.008
−1.576–(−0.439)
<0.001
ULSAM
−0.664
−0.952–(−0.375)
<0.001
PIVUS
−1.076
−1.604–(−0.547)
<0.001 Table 4. Multivariable linear regression models showing the association between standard deviation
increments in eGFR (independent variable, in ml/min/1.73 m2 body surface area (BSA)) and cathepsin-S
(dependent variable, in ng/ml) in all three study cohorts. Multivariable analysis adjusted for age, gender,
body-mass-index and c-reactive protein; the regression coefficient B describes the change of cathepsin-S
(in ng/ml) per change of one standard deviation (SD) of eGFR in every cohort (SD in MCKD 37.2 ml/
min/1.73 m2 BSA, in ULSAM 17.1 ml/min/1.73 m2 BSA, in PIVUS 19.3 ml/min/1.73 m2 BSA). Table 4. Multivariable linear regression models showing the association between standard deviation
increments in eGFR (independent variable, in ml/min/1.73 m2 body surface area (BSA)) and cathepsin-S
(dependent variable, in ng/ml) in all three study cohorts. Multivariable analysis adjusted for age, gender,
body-mass-index and c-reactive protein; the regression coefficient B describes the change of cathepsin-S
(in ng/ml) per change of one standard deviation (SD) of eGFR in every cohort (SD in MCKD 37.2 ml/
min/1.73 m2 BSA, in ULSAM 17.1 ml/min/1.73 m2 BSA, in PIVUS 19.3 ml/min/1.73 m2 BSA). SCIeNTIFIC ReporTS | 7:43538 | DOI: 10.1038/srep43538 6 www.nature.com/scientificreports/ /
p
/ sufficient amounts of the intrinsic Cat-S inhibitor cystatin-C5. We therefore assume tha
levels are largely produced from activated circulating monocytes but not locally in re
i f
h
h
d b
h f
h
ld
d
i
ifi
i i
Figure 2. MCKD study; univariate correlation analysis using Pearson-correlation coe
association between serum cathepsin-S (in ng/ml) and (A) estimated glomerular filtra
EPIcreatinine-cystatin c, in ml/min/1.73 m2 body surface area); (B) serum soluble TNF recep
(C) serum soluble TNF receptor 2 (TNFR-2, in ng/ml). Figure 2. MCKD study; univariate correlation analysis using Pearson-correlation coefficient (r) to assess the
association between serum cathepsin-S (in ng/ml) and (A) estimated glomerular filtration rate (eGFR CKD-
EPIcreatinine-cystatin c, in ml/min/1.73 m2 body surface area); (B) serum soluble TNF receptor 1 (TNFR-1, in ng/ml);
(C) serum soluble TNF receptor 2 (TNFR-2, in ng/ml). sufficient amounts of the intrinsic Cat-S inhibitor cystatin-C5. We therefore assume that in CKD circulating Cat-S
levels are largely produced from activated circulating monocytes but not locally in renal tissue4. This hypothesis
is further strengthened by the fact that we could not detect a significant association between urinary Cat-S/ Discussion Figure 2. MCKD study; univariate correlation analysis using Pearson-correlation coefficient (r) to assess the
association between serum cathepsin-S (in ng/ml) and (A) estimated glomerular filtration rate (eGFR CKD-
EPIcreatinine-cystatin c, in ml/min/1.73 m2 body surface area); (B) serum soluble TNF receptor 1 (TNFR-1, in ng/ml);
(C) serum soluble TNF receptor 2 (TNFR-2, in ng/ml). sufficient amounts of the intrinsic Cat-S inhibitor cystatin-C5. We therefore assume that in CKD circulating Cat-S
levels are largely produced from activated circulating monocytes but not locally in renal tissue4. This hypothesis
is further strengthened by the fact that we could not detect a significant association between urinary Cat-S/ sufficient amounts of the intrinsic Cat-S inhibitor cystatin-C5. We therefore assume that in CKD circulating Cat-S
levels are largely produced from activated circulating monocytes but not locally in renal tissue4. This hypothesis
is further strengthened by the fact that we could not detect a significant association between urinary Cat-S/ sufficient amounts of the intrinsic Cat-S inhibitor cystatin-C5. We therefore assume that in CKD circulating Cat-S
levels are largely produced from activated circulating monocytes but not locally in renal tissue4. This hypothesis
is further strengthened by the fact that we could not detect a significant association between urinary Cat-S/ SCIeNTIFIC ReporTS | 7:43538 | DOI: 10.1038/srep43538 7 www.nature.com/scientificreports/ Study group/Biomarker
regression
coefficient B
95% confidence
interval
p-value
MCKD
Cathepsin-S vs sTNFR-1
1.314
0.833–1.794
<0.001
Cathepsin-S vs sTNFR-2
1.976
1.437–2.514
<0.001
ULSAM
Cathepsin-S vs sTNFR-1
2.474
1.654–3.293
<0.001
PIVUS
Cathepsin-S vs sTNFR-1
4.034
2.297–5.772
<0.001
Cathepsin-S vs sTNFR-2
8.935
7.548–10.323
<0.001
Table 5. Multivariable linear regression models showing the association between (log) sTNFR-1/-2
(independent variable, in ng/ml) and cathepsin-S (dependent variable, in ng/ml) in all three study cohorts. Multivariable analysis adjusted for age, gender, body-mass-index, and c-reactive protein; sTNFR-2 not available
in ULSAM; the regression coefficient B describes the change of cathepsin-S (in ng/ml) per change of (log)
sTNFR-1/2. Study group/Biomarker
regression
coefficient B
95% confidence
interval
p-value
MCKD
Cathepsin-S vs sTNFR-1
1.314
0.833–1.794
<0.001
Cathepsin-S vs sTNFR-2
1.976
1.437–2.514
<0.001
ULSAM
Cathepsin-S vs sTNFR-1
2.474
1.654–3.293
<0.001
PIVUS
Cathepsin-S vs sTNFR-1
4.034
2.297–5.772
<0.001
Cathepsin-S vs sTNFR-2
8.935
7.548–10.323
<0.001
Table 5. Multivariable linear regression models showing the association between (log) sTNFR-1/-2
(independent variable, in ng/ml) and cathepsin-S (dependent variable, in ng/ml) in all three study cohorts. Discussion Multivariable analysis adjusted for age, gender, body-mass-index, and c-reactive protein; sTNFR-2 not available
in ULSAM; the regression coefficient B describes the change of cathepsin-S (in ng/ml) per change of (log)
sTNFR-1/2. Study group/Biomarker
regression
coefficient B
95% confidence
interval
p-value
MCKD
Cathepsin-S vs sTNFR-1
1.314
0.833–1.794
<0.001
Cathepsin-S vs sTNFR-2
1.976
1.437–2.514
<0.001
ULSAM
Cathepsin-S vs sTNFR-1
2.474
1.654–3.293
<0.001
PIVUS
Cathepsin-S vs sTNFR-1
4.034
2.297–5.772
<0.001
Cathepsin-S vs sTNFR-2
8.935
7.548–10.323
<0.001 Table 5. Multivariable linear regression models showing the association between (log) sTNFR-1/-2
(independent variable, in ng/ml) and cathepsin-S (dependent variable, in ng/ml) in all three study cohorts. Multivariable analysis adjusted for age, gender, body-mass-index, and c-reactive protein; sTNFR-2 not available
in ULSAM; the regression coefficient B describes the change of cathepsin-S (in ng/ml) per change of (log)
sTNFR-1/2. Table 5. Multivariable linear regression models showing the association between (log) sTNFR-1/-2
(independent variable, in ng/ml) and cathepsin-S (dependent variable, in ng/ml) in all three study cohorts. Multivariable analysis adjusted for age, gender, body-mass-index, and c-reactive protein; sTNFR-2 not available
in ULSAM; the regression coefficient B describes the change of cathepsin-S (in ng/ml) per change of (log)
sTNFR-1/2. Figure 3. ULSAM study; univariate correlation analysis using Pearson-correlation coefficient (r) to assess the
association between serum cathepsin-S (in ng/ml) and (A) estimated glomerular filtration rate (eGFR CKD-
EPIcystatin c, in ml/min/1.73 m2 body surface area); (B) serum soluble TNF receptor 1 (TNFR-1, in ng/ml). Figure 3. ULSAM study; univariate correlation analysis using Pearson-correlation coefficient (r) to assess the
association between serum cathepsin-S (in ng/ml) and (A) estimated glomerular filtration rate (eGFR CKD-
EPIcystatin c, in ml/min/1.73 m2 body surface area); (B) serum soluble TNF receptor 1 (TNFR-1, in ng/ml). creatinine ratio and eGFR in the ULSAM cohort (Supplementary Figure 1). Furthermore, circulating Cat-S was
not associated with proteinuria/albuminuria in all three cohorts (Supplementary Table 1). Both increased pro-
duction and a decline of glomerular filtration surface, as it occurs along nephron loss during CKD, may reduce
the clearance of Cat-S from the circulation, and therefore, increase circulating Cat-S levels. In fact, Cat-S has a
similar molecular weight to several markers of glomerular filtration such as cystatin C, beta-trace protein, and
beta2-microglobulin. This molecular weight similarity suggests that Cat-S may be freely filtered through the glo-
meruli and thus provide a potential marker of GFR. SCIeNTIFIC ReporTS | 7:43538 | DOI: 10.1038/srep43538 8 www.nature.com/scientificreports/ www.nature.com/scientificreports/ Figure 4. Discussion PIVUS study; univariate correlation analysis using Pearson-correlation coe
association between serum cathepsin-S (in ng/ml) and (A) estimated glomerular filtr
EPIcystatin c, in ml/min/1.73 m2 body surface area); (B) serum soluble TNF receptor 1 (
(C) serum soluble TNF receptor 2 (TNFR-2, in ng/ml). Figure 4. PIVUS study; univariate correlation analysis using Pearson-correlation coefficient (r) to assess the
association between serum cathepsin-S (in ng/ml) and (A) estimated glomerular filtration rate (eGFR CKD-
EPIcystatin c, in ml/min/1.73 m2 body surface area); (B) serum soluble TNF receptor 1 (TNFR-1, in ng/ml);
(C) serum soluble TNF receptor 2 (TNFR-2, in ng/ml). Markers of systemic inflammation, such as urinary monocyte chemotactic protein-1, tumor necrosis
factor-alpha (TNF-α), and interleukin-6 (IL-6) have been shown to correlate with the severity of CKD28,29. In
addition, inflammatory markers sTNFR1 and sTNFR2 have been associated with CKD26. The latter markers
are not only biomarkers of systemic inflammation but also predict cardiovascular morbidity and mortality13. SCIeNTIFIC ReporTS | 7:43538 | DOI: 10.1038/srep43538 9 www.nature.com/scientificreports/ Both TNFRs are shed mainly by activated endothelial cells and are therefore considered to represent markers of
inflammation-related endothelial dysfunction30. Other markers of endothelial dysfunction, such as asymmetric
dimethylarginine (ADMA), L-arginine, soluble vascular adhesion molecule-1 (sVCAM-1) and soluble E-selectin
(sE-selectin) have also been associated with CKD31, demonstrating that endothelial dysfunction contributes to
cardiovascular morbidity/mortality in CKD patients. We have recently shown that Cat-S is an upstream inducer
of inflammation-related endothelial dysfunction by cleaving the extracellular domain of the protease-activated
receptor (PAR)-2 on the luminal membranes of vascular endothelial cells5. As a working hypothesis, any stimulus
that activates monocytes and neutrophils in the circulation, as may occur during extra-corporal circulation dur-
ing hemodialysis or by bacterial endotoxin leakage from the intestinal tract32, may induce Cat-S release, which
activates endothelial cells to release sTNFR1 and sTNFR2. In line with this hypothesis Cat-S has been previously
shown to be associated with the sTNFRs in hemodialysis patients, a highly inflammatory milieu33. The vasculo-
pathic effects of Cat-S also have a direct role in promoting cardiovascular complications, such as the processes of
atherogenesis and vascular wall degeneration, including aneurysm formation and media calcification6–12,18,19,34.f g
g
g
yi
Interestingly, we observed that Cat-S and sTNFR1/2 concentrations differed between the three human cohorts
in groups with similar eGFR (e.g. CKD III°). Therefore we wondered if other demographic/laboratory parameters
besides kidney function might influence their blood concentrations. Discussion We reran an exemplary multivariable linear
regression analysis focusing on parameters possibly influencing Cat-S in MCKD. We set Cat-S as the dependent
variable and added independent variables as follows: eGFR, gender, age, BMI, total serum protein, serum CRP
and prevalence of diabetes mellitus, arterial hypertension, vascular disease and tumor disease. In this analy-
sis, besides eGFR only gender was weakly associated with Cat-S (p = 0.043), otherwise no significant data was
obtained. Since the clinical samples were similar, we surmise that methodological rather than clinical circum-
stances lead to these findings. In particular, the sample material may have an influence on the results since plasma
was used in MCKD and serum in PIVUS/ULSAM. Furthermore, samples were stored for a much longer period of
time in PIVUS/ULSAM. Currently there are no reference values for Cat-S or sTNFR 1/2 in human material, and it
would be important to determine in the future whether different value ranges exist for different sample materials
and storage conditions. g
Our study has several strengths, including the use of both experimental and clinical data and the validation
of results in independent clinical cohorts with a wide range of glomerular filtration rate. One limitation is that
statistical analyses were not adjusted for multiple testing. However, the consistency of results from three different
cohorts and a mouse model would argue against spurious association as an explanation of our findings. Other
limitations include the lack of longitudinal data in the human cohorts and the use of different sample material
in the three human cohorts, possibly limiting the comparability of the results obtained in the three cohorts. It is
acknowledged that our data are cross-sectional in nature and only reflect the association between Cat-S and kid-
ney function, with no outcome data provided. Hence, no clinical consequences can be drawn from our findings. However, the association of circulatory Cat-S with outcomes in CKD patients should be evaluated in future epi-
demiological studies. Lastly, additional markers of endothelial dysfunction and systemic inflammation that have
been shown to be associated with CKD could not be measured in our cohorts, and might have been compared
with Cat-S to validate the proinflammatory status. pl
y
In summary, Cat-S, a cysteine protease released by activated immune cells, is consistently found at increased
levels that correlate with the progression of CKD in mice and humans. Discussion It is tempting to speculate that Cat-S is
a causative factor in this association, and a mediator of CKD-related cardiovascular morbidity and mortality. As pharmacological Cat-S inhibition has been shown to prevent macrovascular and microvascular disease in
experimental animal models, there appears to be a valid scientific rationale to study whether Cat-S is associated
with adverse outcomes in CKD and whether its inhibition can reduce cardiovascular morbidity and mortality in
CKD patients. References 1. Reiser, J., Adair, B. & Reinheckel, T. Specialized roles for cysteine cathepsins in health and disease. J Clin Invest. 120, 3421–3431
(2010). 2. Rupanagudi, K. V. et al. Cathepsin S inhibition suppresses systemic lupus erythematosus and lupus nephritis because cathepsi
essential for MHC class II-mediated CD4 T cell and B cell priming. Ann Rheum Dis. 74, 452–463 (2015). p
g
3. Shi, G. P. et al. Cathepsin S required for normal MHC class II peptide loading and germinal center development. Immunity. 10,
197–206 (1999).l 4. Liuzzo, J. P., Petanceska, S. S., Moscatelli, D. & Devi, L. A. Inflammatory mediators regulate cathepsin S in macrophages and
microglia: A role in attenuating heparan sulfate interactions. Mol Med. 5, 320–333 (1999). g
g
p
5. Kumar V. S. et al. Cathepsin S Cleavage of Protease-Activated Receptor-2 on Endothelial Cells Promotes Microvascular Diabetes
Complications. J Am Soc Nephrol. 27, 1635–1649 (2016).ii 6. Aikawa, E. et al. Arterial and aortic valve calcification abolished by elastolytic cathepsin S deficiency in chronic renal disease
Circulation 119, 1785–1794 (2009). 7. Lafarge, J. C., Naour, N., Clement, K. & Guerre-Millo, M. Cathepsins and cystatin C in atherosclerosis and obesity. Biochimie 92
1580–1586 (2010).h 8. Liu, J. et al. Lysosomal cysteine proteases in atherosclerosis. Arterioscler Thromb Vasc Biol. 24, 1359–1366 (2004).i l. Lysosomal cysteine proteases in atherosclerosis. Arterioscler Thro 9. Qin, Y. et al. Deficiency of cathepsin S attenuates angiotensin II-induced abdominal aortic aneurysm formation in apolipoprotein
E-deficient mice. Cardiovasc Res. 96, 401–410 (2012).h i
10. Rodgers, K. J. et al. Destabilizing role of cathepsin S in murine atherosclerotic plaques. Arterioscler Thromb Vasc Biol. 26, 851–856
(2006). 11. Samokhin, A. O., Lythgo, P. A., Gauthier, J. Y., Percival, M. D. & Bromme, D. Pharmacological inhibition of cathepsin S decreases
atherosclerotic lesions in Apoe-/- mice. J Cardiovasc Pharmacol. 56, 98–105 (2010).ii p
2. Sukhova, G. K. et al. Deficiency of cathepsin S reduces atherosclerosis in LDL receptor-deficient mice. J Clin Invest. 111, 897–906
(2003). (
)
13. Jobs, E. et al. Association between serum cathepsin S and mortality in older adults. JAMA. 306, 1113–1121 (2011). 14. Jobs, E. et al. Serum cathepsin S is associated with decreased insulin sensitivity and the development of type 2 diabetes in a (
)
13. Jobs, E. et al. Association between serum cathepsin S and mortality in older adults. JAMA. 306, 1113–1121 (2011). 14. Jobs, E. et al. www.nature.com/scientificreports/ Estimating glomerular filtration rate from serum creatinine and cystatin C. N Engl J Med. 367 20–29 (2012). ,
g gi
y
g J
(
)
26. Carlsson, A. C., Larsson, T. E., Helmersson-Karlqvist, J., Larsson, A., Lind, L. & Arnlov, J. Soluble TNF receptors and kidney
dysfunction in the elderly. J Am Soc Nephrol. 25, 1313–1320 (2014). 26. Carlsson, A. C., Larsson, T. E., Helmersson-Karlqvist, J., Larsson, A., Lind, L. & Arnlov, J
dysfunction in the elderly. J Am Soc Nephrol. 25, 1313–1320 (2014). y
y
p
27. Cochat, P. & Rumsby, G. Primary hyperoxaluria. N Engl J Med. 369, 649–658 (2013). 28. Lee, B. T. et al. Association of C-reactive protein, tumor necrosis factor-alpha, and interleukin-6 with chronic kidney disease. BMC
Nephrol. 16, 77 (2015).lh 29. Nadkarni, G. N. et al. Association of Urinary Biomarkers of Inflammation, Injury, and Fibrosis with Renal Function Decline
ACCORD Trial. Clin J Am Soc Nephrol. 11, 1343–1352 (2016). 30. Mikacenic, C. et al. Biomarkers of Endothelial Activation Are Associated with Poor Outcome in Critical Illness. PLoS One. 10,
e0141251 (2015). 31. Chen, J. et al. Interrelationship of Multiple Endothelial Dysfunction Biomarkers with Chronic Kidney Disease. PLoS One. 10,
e0132047 (2015).h (
)
2. Anders, H. J., Andersen, K. & Stecher, B. The intestinal microbiota, a leaky gut, and abnormal immunity in kidney disease. Kidney
Int. 83, 1010–1016 (2013).hl ,
(
)
3. Carlsson, A. C. et al. Endostatin, Cathepsin S, and Cathepsin L, and Their Association with Inflammatory Markers and Mortality in
d
d l
l
d
f
(
) 33. Carlsson, A. C. et al. Endostatin, Cathepsin S, and Cathepsin L, and Their Association with Inflammatory Markers and Morta
Patients Undergoing Hemodialysis. Blood Purif. 39, 259–265 (2015).i g
g
y
f
,
(
)
34. Shi, G. P. et al. Deficiency of the cysteine protease cathepsin S impairs microvessel growth. Circ Res. 92, 493–500 (2003). Acknowledgements g
We thank Dr. Paul Albert, PhD, University of Ottawa for revision of the manuscript. HJ.A. was supported by
European Union’s Horizon 2020 research and innovation programme under grant agreement No. 668036 (project
RELENT). A.V. and J.Ä. were supported by grants from the Swedish Research Council, Swedish Heart-Lung
foundation, the European Union Horizon 2020 (Project SCOPE, grant number 634869), the Marianne and
Marcus Wallenberg Foundation, Dalarna University, and Uppsala University. This work was supported by the
German Research Foundation (DFG) and the Technische Universität München within the funding programme
Open Access Publishing. h
German Research Foundation (DFG) and the Technische Universität München within the funding programm
Open Access Publishing. www.nature.com/scientificreports/ www.nature.com/scientificreports/ 5. Naour, N. et al. Cathepsins in human obesity: changes in energy balance predominantly affect cathepsin s in adipose tissue and in
circulation. J Clin Endocrinol Metab. 95, 1861–1868 (2010). 6. Jobs, E. et al. Serum cathepsin S is associated with serum C-reactive protein and interleukin-6 independently of obesity in elderly
men. J Clin Endocrinol Metab. 95, 4460–4644 (2010). 17. Lv, B. J., Lindholt, J. S., Cheng, X., Wang, J. & Shi, G. P. Plasma cathepsin S and cystatin C levels and risk of abdominal aortic
aneurysm: a randomized population-based study. PLoS One. 7, e41813 (2012). y
p p
y
8. Lutgens, S. P., Cleutjens, K. B., Daemen, M. J. & Heeneman, S. Cathepsin cysteine proteases in cardiovascular disease. FASEB J. 21
3029–3041 (2007).i 19. Figueiredo, J. L. et al. Selective cathepsin S inhibition attenuates atherosclerosis in apolipoprotein E-deficient mice with chronic
renal disease. Am J Pathol. 185, 1156–1166 (2015). 20. Go, A. S., Chertow, G. M., Fan, D., McCulloch, C. E. & Hsu, C. Y. Chronic kidney disease and the risks of death, cardiovascular
events, and hospitalization. N Engl J Med. 351, 1296–1305 (2004). p
g
1. Stenvinkel, P. et al. Emerging biomarkers for evaluating cardiovascular risk in the chronic kidney disease patient: how do new piece
fit into the uremic puzzle? Clin J Am Soc Nephrol. 3, 505–521 (2008). g
21. Stenvinkel, P. et al. Emerging biomarkers for evaluating cardiovascular risk in the chronic kidne
fit into the uremic puzzle? Clin J Am Soc Nephrol. 3, 505–521 (2008). 21. Stenvinkel, P. et al. Emerging biomarkers for evaluating cardiov i
22. Mulay, S. R. et al. Oxalate-induced chronic kidney disease with its uremic and cardiovascular complications in C57BL/6 mice. Am J
Physiol Renal Physiol. 2016 Jan 13: ajprenal.00488.2015 (epub ahead of print). 23. Schreiber, A. et al. Transcutaneous measurement of renal function in conscious mice. Am J Physiol Renal Physiol. 303, F783
(2012). 24. Stevens, P. E., Levin, A., Kidney Disease: Improving Global Outcomes Chronic Kidney Disease Guideline Development Work
Group. Evaluation and management of chronic kidney disease: synopsis of the kidney disease: improving global outcomes 2012
clinical practice guideline. Ann Intern Med. 158, 825–830 (2013).i y
p
g
y
p
Group. Evaluation and management of chronic kidney disease: synopsis of the kidney disease: improving global outcomes 2012
clinical practice guideline. Ann Intern Med. 158, 825–830 (2013).i p
g
,
(
)
25. Inker, L. A. et al. Author Contributions Conception or design, or analysis and interpretation of data, or both: D.S., S.V.K., M.T., S.R.M., A.C.C., J.Ä.,
H.J.A. Drafting the article or revising it: D.S., S.V.K., A.L., L.L., U.R., A.C.C., J.Ä., H.J.A. Providing intellectual
content of critical importance to the work described: A.L., L.L., U.R., L.R., U.H., A.C.C., J.Ä., H.J.A. Final approval
of the version to be published: all authors. References Serum cathepsin S is associated with decreased insulin sensitivity and the development of type 2 diabetes in a
community-based cohort of elderly men. Diabetes Care. 36, 163–165 (2013). 13. Jobs, E. et al. Association between serum cathepsin S and mortality in older adults. JAMA. 306, 1113–1121 (2011). J b
E
l S
h
i
S i
i
d
i h d
d i
li
i i i
d h d
l
f
di b
i 13. Jobs, E. et al. Association between serum cathepsin S and mort SCIeNTIFIC ReporTS | 7:43538 | DOI: 10.1038/srep43538 10 Additional Information Supplementary information accompanies this paper at http://www.nature.com/srepihi Competing financial interests: The authors declare no competing financial interests. Competing financial interests: The authors declare no competing financial interests. How to cite this article: Steubl, D. et al. Circulating cathepsin-S levels correlate with GFR decline and sTNF
and sTNFR2 levels in mice and humans. Sci. Rep. 7, 43538; doi: 10.1038/srep43538 (2017). How to cite this article: Steubl, D. et al. Circulating cathepsin-S levels correlate with GFR decline and sTNFR1
nd sTNFR2 levels in mice and humans. Sci. Rep. 7, 43538; doi: 10.1038/srep43538 (2017). Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and
institutional affiliations. Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps an
institutional affiliations. This work is licensed under a Creative Commons Attribution 4.0 International License. The images
or other third party material in this article are included in the article’s Creative Commons license,
unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license,
users will need to obtain permission from the license holder to reproduce the material. To view a copy of this
license, visit http://creativecommons.org/licenses/by/4.0/ SCIeNTIFIC ReporTS | 7:43538 | DOI: 10.1038/srep43538 11
|
https://openalex.org/W2938102902
|
https://europepmc.org/articles/pmc6515182?pdf=render
|
English
| null |
High Accuracy Buoyancy for Underwater Gliders: The Uncertainty in the Depth Control
|
Sensors
| 2,019
|
cc-by
| 12,008
|
Received: 8 March 2019; Accepted: 12 April 2019; Published: 17 April 2019
Received: 8 March 2019; Accepted: 12 April 2019; Published: date Received: 8 March 2019; Accepted: 12 April 2019; Published: 17 April 2019
Received: 8 March 2019; Accepted: 12 April 2019; Published: date Abstract: This paper is a section of several preliminary studies of the Underwater Drones Group
of the Università degli Studi “Roma Tre” Science Department: We describe the study philosophy,
the theoretical technological considerations for sizing and the development of a technological
demonstrator of a high accuracy buoyancy and depth control. We develop the main requirements and
the boundary conditions that design the buoyancy system and develop the mathematical conditions
that define the main parameters. Abstract: This paper is a section of several preliminary studies of the Underwater Drones Group of
the Università degli Studi “Roma Tre” Science Department: We describe the study philosophy, the
theoretical technological considerations for sizing and the development of a technological
demonstrator of a high accuracy buoyancy and depth control. We develop the main requirements
and the boundary conditions that design the buoyancy system and develop the mathematical conditions
that define the main parameters. Keywords: uncertainty; buoyancy; depth control; accuracy; AUV; glider; autonomous; underwater;
vehicle
Keywords: uncertainty; buoyancy; depth control; accuracy; AUV; glider; autonomous; underwater;
vehicle sensors sensors sensors High Accuracy Buoyancy for Underwater Gliders:
The Uncertainty in the Depth Control †
Article
High Accuracy Buoyancy for Underwater Gliders:
The Uncertainty in the Depth Control † Enrico Petritoli, Fabio Leccese *
and Marco Cagnetti
Enrico Petritoli Fabio Leccese * and Marco Cagnetti Enrico Petritoli, Fabio Leccese *
and Marco Cagnetti
Enrico Petritoli Fabio Leccese * and Marco Cagnetti Science Department, Università degli Studi “Roma Tre”, Via della Vasca Navale n. 84, 00146 Rome, Italy;
e_petritoli@libero.it (E.P.); ing.marco.cagnetti@gmail.com (M.C.)
* Correspondence: fabio.leccese@uniroma3.it; Tel.: +39-06-5733-7347
† This paper is an extended version of our paper published in Petritoli, E.; Leccese, F; Cagnetti, M. A High
Accuracy Buoyancy System Control for an Underwater Glider. In Proceedings of the second IEEE
International Workshop on Metrology for the Sea (MetroSea), Bari, Italy, 8–10 October 2018. g
Science Department, Università degli Studi “Roma Tre”, Via della Vasca Navale n. 84, 00146 Rome, Italy;
e_petritoli@libero.it (E.P.); ing.marco.cagnetti@gmail.com (M.C.)
* Correspondence: fabio.leccese@uniroma3.it; Tel.: +39-06-5733-7347
† This paper is an extended version of our paper published in Petritoli, E.; Leccese, F; Cagnetti, M. A High
Accuracy Buoyancy System Control for an Underwater Glider. In Proceedings of the second IEEE
International Workshop on Metrology for the Sea (MetroSea), Bari, Italy, 8–10 October 2018. Science Department, Università degli Studi “Roma Tre”, Via della Vasca Navale n. 84, 00146 Rome, Italy;
e_petritoli@libero.it (E.P.); ing.marco.cagnetti@gmail.com (M.C.)
g
Science Department, Università degli Studi “Roma Tre”, Via della Vasca Navale n. 84, 00146 Rome, Italy;
e petritoli@libero it (E P ); ing marco cagnetti@gmail com (M C ) Science Department, Università degli Studi “Roma Tre”, Via della Vasca Navale n. 84, 00146 Rome, Italy;
e_petritoli@libero.it (E.P.); ing.marco.cagnetti@gmail.com (M.C.)
* Correspondence: fabio.leccese@uniroma3.it; Tel.: +39-06-5733-7347
† This paper is an extended version of our paper published in Petritoli, E.; Leccese, F; Cagnetti, M. A High
Accuracy Buoyancy System Control for an Underwater Glider. In Proceedings of the second IEEE
International Workshop on Metrology for the Sea (MetroSea), Bari, Italy, 8–10 October 2018. Science Department, Università degli Studi “Roma Tre”, Via della Vasca Navale n. 84, 00146 Rome, Italy;
e_petritoli@libero.it (E.P.); ing.marco.cagnetti@gmail.com (M.C.)
* Correspondence: fabio.leccese@uniroma3.it; Tel.: +39-06-5733-7347
† This paper is an extended version of our paper published in Petritoli, E.; Leccese, F; Cagnetti, M. A High
Accuracy Buoyancy System Control for an Underwater Glider. In Proceedings of the second IEEE
International Workshop on Metrology for the Sea (MetroSea) Bari Italy 8–10 October 2018 www.mdpi.com/journal/sensors
www.mdpi.com/journal/sensors
1.1.3. The Mk. III Architecture The Mk. III sub-glider has a cylindrical fuselage with a radome on the bow containing the
customizable payload and, on the other end, the hydrodynamic fairing. The vehicle does not have
moving surfaces: control is provided by the displacement of the battery package that varies the position
of the centre of mass. The wings aerofoil is based on the Eppler E838 Hydrofoil. The aerofoil has the
maximum thickness 18.4% at 37.2% chord and maximum camber 0% at 46.5% chord. The arrangement
of the internal sectors is visible in Figure 2a,b. The buoyancy system is contained in the buoyancy
control bay: it accommodates the buoyancy motor and the oil tank and provides longitudinal balance
to the system by adjusting the level in the reservoir. The bladder is contained in the hydrodynamic
fairing, in contact with the open water. The fairing is not a critical structural part—it has the task of not
disturbing the hydrodynamic flow of the fuselage [28]. 1.1.1. AUV Evolution The exploration of the underwater world has always been one of mankind’s dreams: submarines
and bathyscaphes (for extreme depths) have been developed to study the “deep blue”. Due to obvious
dangers, the human exploration can take place only for very short periods and very limited areas: for
these reasons, the exploration of the sea has immediately been drawn towards unmanned automatic
systems [6–8]. An AUV is a vehicle that travels underwater without requiring input from an operator; this means
that it must be equipped with a “brain” that regulates and coordinates its position, its depth and its
speed: moreover, it is able to collect and store data from the payload. One of the first realizations was
the Autonomous LAgrangian Circulation Explorer (ALACE) system, a buoy that was able to vary its
buoyancy and therefore its depth. Although it possessed a great endurance, it only could be employed
for great depths and in open sea—the consequences of these limitations are evident. The next step was the use of Remote Operated Vehicles (ROVs). These, thanks to the constant
development of electronic miniaturization, are extremely high performing vehicles for short-lasting
marine operations, but the require the constant presence of a support vessel. The need to get rid of the randomness of the currents has led to the natural development of the
underwater glider concept [9–15]. 1.1.2. The Underwater Glider An underwater glider is a vehicle that, by changing its buoyancy, moves up and down in the
ocean like a profiling float [16]. It uses hydrodynamic wings to convert vertical motion into horizontal
motion, moving forward with very low power consumption [17–22]. While not as fast as conventional
AUVs, the glider, using buoyancy-based propulsion, offers increased range and endurance compared
to motor-driven vehicles and missions may extend to months and to several thousands of kilometres
in range. An underwater glider follows an up-and-down, sawtooth-like mission profile providing data
on temporal and spatial scales unavailable with previous types of AUVs [23–27]. 1. Introduction
1. Introduction This paper is part of several preliminary studies by Underwater Drones Group (UDG) of the
Science Department of the Università degli Studi “Roma Tre”, which is developing an advanced
Autonomous Underwater Vehicle (AUV) for the exploration of the sea at high depths. The final aim
of the project is to create a platform for underwater scientific research that can accommodate a wide
range of different payloads. This paper is part of several preliminary studies by Underwater Drones Group (UDG) of the
Science Department of the Università degli Studi “Roma Tre”, which is developing an advanced
Autonomous Underwater Vehicle (AUV) for the exploration of the sea at high depths. The final aim
of the project is to create a platform for underwater scientific research that can accommodate a wide
range of different payloads. We will examine the buoyancy system and evaluate its sizing; then we will illustrate the
technological solution we have come up with in order to realize the hydraulic system to be assembled
in the Underwater Glider Mk. III (see Figure 1) [1–5]. range of different payloads. We will examine the buoyancy system and evaluate its sizing; then we will illustrate the
technological solution we have come up with in order to realize the hydraulic system to be assembled
in the Underwater Glider Mk. III (see Figure 1) [1–5]. (a)
(b)
Figure 1. Perspective view of the Underwater Glider Mk. III. (a) Rear/port; (b) front/starboard. Figure 1. Perspective view of the Underwater Glider Mk. III. (a) Rear/port; (b) front/starboard. (b) (b) (b) (a) Figure 1 Pe specti e
ie
of the U de
ate Glide Mk III (a) Rea /po t (b) f o t/sta boa d
Figure 1. Perspective view of the Underwater Glider Mk. III. (a) Rear/port; (b) front/starboard. www.mdpi.com/journal/sensors
www.mdpi.com/journal/sensors Sensors 2019, 19, x; doi:
Sensors 2019, 19, 1831; doi:10.3390/s19081831 2 of 17 Sensors 2019, 19, 1831 1.1. The Underwater Glider where ζ is the parameter vector [29].
,
, (b) (a) Figure 2. Underwater Glider Mk. III cutaway: (a) fuselage prospective section; (b) fuselage sagittal
Figure 2. Underwater Glider Mk. III cutaway: (a) fuselage prospective section; (b) fuselage
sagittal section. −𝜁௬
𝜁௫
0
here 𝜁 is the parameter vector [29]. al conventions and symbols that will b
d the N axis. d the Z axis. nd the X axis. −𝜁௭
𝜁௬
0
−𝜁௫
𝜁
0
ቍ
(b) .4. Conventions
We introduce, for clarity, the mathematical conventions and symbols that will be used in
bsequent discussion (see Figure 3a,b):
here:
α (or φ) is the angle between the x axis and the N axis. β (or θ) is the angle between the z axis and the Z axis. γ (or ψ) is the angle between the N axis and the X axis. For the rotation matrix, we have:
𝑹= ቌ
0
−𝜁௭
𝜁௬
𝜁௭
0
−𝜁௫
−𝜁௬
𝜁௫
0
ቍ
(1
here 𝜁 is the parameter vector [29]. (a)
(b)
Figure 3. The Euler angles. (a) Body frame (blue) and reference frame (red); (b) The body frame
referred to the drone. Figure 3. The Euler angles. (a) Body frame (blue) and reference frame (red); (b) The body frame
referred to the drone. ions
duce, for clarity, the mathematic
scussion (see Figure 3a,b):
s the angle between the x axis an
the angle between the z axis and
s the angle between the N axis an
tation matrix, we have:
𝑹= ቌ
0
𝜁௭
𝜁
(a) (b) (b) (a) 𝜁௬
𝜁௫
0
ere 𝜁 is the parameter vector [29]. Figure 3. The Euler angles. (a) Body frame (blue) and reference frame (red); (b) The body frame
referred to the drone
Figure 3. The Euler angles. (a) Body frame (blue) and reference frame (red); (b) The body frame
referred to the drone. where ζ is the parameter vector [29].
,
, where ζ is the parameter vector [29]. (a)
(b)
Figure 2. Underwater Glider Mk. III cutaway: (a) fuselage prospective section; (b) fuselage sagittal
Figure 2. Underwater Glider Mk. III cutaway: (a) fuselage prospective section; (b) fuselage
sagittal section. (a)
(b)
Figure 2. Underwater Glider Mk. III cutaway: (a) fuselage prospective section; (b) fuselage sagittal
section. 1.1.4. Conventions
We introduce, for clarity, the mathematical conventions and symbols that will be used in the
subsequent discussion (see Figure 3a,b):
where:
•
α (or φ) is the angle between the x axis and the N axis. •
β (or θ) is the angle between the z axis and the Z axis. •
γ (or ψ) is the angle between the N axis and the X axis. For the rotation matrix, we have:
𝑹= ቌ
0
−𝜁௭
𝜁௬
𝜁௭
0
−𝜁௫
−𝜁௬
𝜁௫
0
ቍ
(1)
Where 𝜁 is the parameter vector [29]. (a)
(a)
Figure 2. Underwater Glider Mk. III cutaway: (a) f
section. 1.1.4. Conventions
We introduce, for clarity, the mathematical co
subsequent discussion (see Figure 3a,b):
where:
•
α (or φ) is the angle between the x axis and the
•
β (or θ) is the angle between the z axis and the
•
γ (or ψ) is the angle between the N axis and th
For the rotation matrix, we have:
𝑹= ቌ
0
−𝜁௭
𝜁௭
0 (b)
(b)
uselage prospective section; (b) fuselage sagittal
nventions and symbols that will be used in th
N axis. Z axis. e X axis. 𝜁௬
−𝜁௫
0
ቍ
(1) (a)
(b)
Figure 2. Underwater Glider Mk. III cutaway: (a) fuselage prospective section; (b) fuselage sagittal
Figure 2. Underwater Glider Mk. III cutaway: (a) fuselage prospective section; (b) fuselage
sagittal section. (a)
(b)
Figure 2. Underwater Glider Mk. III cutaway: (a) fuselage prospective section; (b) fuselage sagittal
section. 1.1.4. Conventions
We introduce, for clarity, the mathematical conventions and symbols that will be used in th
subsequent discussion (see Figure 3a,b):
where:
•
α (or φ) is the angle between the x axis and the N axis. •
β (or θ) is the angle between the z axis and the Z axis. •
γ (or ψ) is the angle between the N axis and the X axis. For the rotation matrix, we have:
𝑹= ቌ
0
−𝜁௭
𝜁௬
𝜁௭
0
−𝜁௫
−𝜁௬
𝜁௫
0
ቍ
(1)
Where 𝜁 is the parameter vector [29]. 2. Materials and Methods 2.1. The Buoyancy System 1.1.4. Conventions We introduce, for clarity, the mathematical conventions and symbols that will be used in the
subsequent discussion (see Figure 3a,b): where: •
α (or ϕ) is the angle between the x axis and the N axis. •
β (or θ) is the angle between the z axis and the Z axis. •
γ (or ψ) is the angle between the N axis and the X axis. 3 of 17 Sensors 2019, 19, 1831 For the rotation matrix, we have: For the rotation matrix, we have: For the rotation matrix, we have: R =
0
−ζz
ζy
ζz
0
−ζx
−ζy
ζx
0
R =
0
−ζz
ζy
ζz
0
−ζx
−ζy
ζx
0
(1)
3 of 17 (1) 2 1 2 The System Prototype
2.1.2. The System Prototype y
yp
Our group has developed a technology demonstrator (see Figure 4) of the buoyancy system to
validate the related technology and then to test it. To reduce the force required to actuate the oil
piston, which pushes the oil in the bladder at high pressure, is necessary to reduce the piston surface
(diameter) and increase the stroke: so, the buoyancy engine resembles a “shotgun”. An open-loop
stepper motor was used to drive the screw inside the actuator that, in turn, pushes the piston. Two
l
id
l
l t th fl
f il i t th bl dd
[53 57]
Our group has developed a technology demonstrator (see Figure 4) of the buoyancy system to
validate the related technology and then to test it. To reduce the force required to actuate the oil piston,
which pushes the oil in the bladder at high pressure, is necessary to reduce the piston surface (diameter)
and increase the stroke: so, the buoyancy engine resembles a “shotgun”. An open-loop stepper motor
was used to drive the screw inside the actuator that, in turn, pushes the piston. Two solenoid valves
regulate the flow of oil into the bladder [53–57]. solenoid valves regulate the flow of oil into the bladder [53–57]. The first problem was the occurrence of actuator buckling: under the push of the engine, the
probability of a part bending is high, thus deforming the thread and jamming the mechanism. The
problem was solved by constructing a rigid cage with four struts that support the piston's push load,
leaving the screw only with the rolling friction load. In the early project development stages, the
workgroup was oriented to use a centrifugal pump for all drives: this technology however did not
allow us to create strong pressure differences; the need to use a more powerful engine was also
highlighted because the prevalence was too low: this would have led to an excessive battery
consumption. The second solution was to use volumetric pumps in order to obtain greater differences
The first problem was the occurrence of actuator buckling: under the push of the engine,
the probability of a part bending is high, thus deforming the thread and jamming the mechanism. The problem was solved by constructing a rigid cage with four struts that support the piston’s push
load, leaving the screw only with the rolling friction load. 2.1.1. Basic Concepts Gliders are controlled through hydrostatics (vertical forces) and manipulate hydrostatic balances
in order to accomplish roll and pitch of the vehicle. Stability of the vehicle is a major critical factor: 4 of 17
static Sensors 2019, 19, 1831
2.1.1. Basic Concep
Glid a stable vehicle has the centre of gravity below the centre of buoyancy. In this configuration, the weight
of the vehicle creates a restoring moment to add stability to the vehicle. Roll and pitch on the glider
is accomplished by moving the battery pack. Figure 4 below displays a basic concept of a buoyancy
system for the glider [30–36]. balances in order to accomplish roll and pitch of the vehicle. Stability of the vehicle is a major critical
factor: a stable vehicle has the centre of gravity below the centre of buoyancy. In this configuration,
the weight of the vehicle creates a restoring moment to add stability to the vehicle. Roll and pitch on
the glider is accomplished by moving the battery pack. Figure 4 below displays a basic concept of a
buoyancy system for the glider [30–36]. Figure 4. Basic scheme of the buoyancy system. Figure 4. Basic scheme of the buoyancy system. Figure 4. Basic scheme of the buoyancy system. Figure 4. Basic scheme of the buoyancy system. The system is extremely simple: while descending, hydraulic fluid moves from the external
inflatable bladder, which produces a high pressure in the internal reservoir, which is at a low pressure
through a valve: the decrease in volume of the bladder creates an increase in density, causing negative
buoyancy [37–44]
The system is extremely simple: while descending, hydraulic fluid moves from the external
inflatable bladder, which produces a high pressure in the internal reservoir, which is at a low pressure
through a valve: the decrease in volume of the bladder creates an increase in density, causing negative
buoyancy [37–44]. buoyancy [37–44]. While ascending, hydraulic fluid moves from internal accumulator to the external inflatable
bladder through the pump. The increase in volume creates a decrease in density causing positive
buoyancy. The seawater also flushes out the open hydrodynamic fairing of the vehicle, aiding it to
rise to the surface. For neutral buoyancy, the vehicle must have a density equal to seawater [45–52]. While ascending, hydraulic fluid moves from internal accumulator to the external inflatable
bladder through the pump. The increase in volume creates a decrease in density causing positive
buoyancy. 2.1.1. Basic Concepts The seawater also flushes out the open hydrodynamic fairing of the vehicle, aiding it to rise
to the surface. For neutral buoyancy, the vehicle must have a density equal to seawater [45–52]. 2 1 2 The System Prototype
2.1.2. The System Prototype In the early project development stages, the
workgroup was oriented to use a centrifugal pump for all drives: this technology however did not allow
us to create strong pressure differences; the need to use a more powerful engine was also highlighted
because the prevalence was too low: this would have led to an excessive battery consumption. The
second solution was to use volumetric pumps in order to obtain greater differences in pressure (even
ones considerably higher than needed). Unfortunately, these would require too much power and are
too heavy for our small vehicle [58–62]. At this stage of development, we have also thought to use the oil tank only as a passive fluid
reservoir and trimmable counterweight of the payload and as an active actuator for longitudinal
stability. The long travel of the piston (forward or backward) ensures the necessary variation of the
bladder volume for manoeuvrability [63]. 5 of 17
tudinal
n of the
udinal
of the Sensors 2019, 19, 1831
reservoir and trim
bili
Th l
reservoir and trim
stability The long Now, the system prototype seems to work correctly and promises new developments: it is under
in several fatigue cycle trials in order to investigate the parts more prone to failure as a sort of burn-in
test. Our prototype (breadboard) is presented in Figure 5. bladder volume for manoeuvrability [63]. Now, the system prototype seems to work correctly and promises new developments: it is under
in several fatigue cycle trials in order to investigate the parts more prone to failure as a sort of burn-
in test. Our prototype (breadboard) is presented in Figure 5. y [
]
Now, the system prototype seems to work correctly and promises new developments: it is under
in several fatigue cycle trials in order to investigate the parts more prone to failure as a sort of burn-
in test. Our prototype (breadboard) is presented in Figure 5. Figure 5. Buoyancy engine prototype. Figure 5. Buoyancy engine prototype. Figure 5. Buoyancy engine prototype. Figure 5. Buoyancy engine prototype. Figure 5. Buoyancy engine prototype. Figure 5. Buoyancy engine prototype. 2.2. Buoyancy
2.2. Buoyancy
2.2. Buoyancy y
y
Archimedes' principle is the main concept underlying the buoyancy of underwater vehicles. When a vehicle is submerged in water, a buoyant force acts on the body vertically upward due to the
pressure forces below the submerged body being greater than the pressure forces above. The buoyant
force results in a value equal to the weight it displaces [64]. Archimedes’ principle is the main concept underlying the buoyancy of underwater vehicles. When a vehicle is submerged in water, a buoyant force acts on the body vertically upward due to the
pressure forces below the submerged body being greater than the pressure forces above. The buoyant
force results in a value equal to the weight it displaces [64]. Archimedes' principle is the main concept underlying the buoyancy of underwater vehicles. When a vehicle is submerged in water, a buoyant force acts on the body vertically upward due to the
pressure forces below the submerged body being greater than the pressure forces above. The buoyant
force results in a value equal to the weight it displaces [64]. BBB = Buoyancy of the oil bladder. BGB = Buoyancy of the naked glider. 2.2.2. Dynamic Balance on the Vertical Plane
2.2.2. Dynamic Balance on the Vertical Plane Here, the drone dive (or emersion) is examined at constant speed: it is in steady-state gliding,
the geometry of total forces is explained in Figure 7a. Figure 7b shows the Eppler E838 characteristic
“Cl vs. Alpha (=angle of attack αatt)”. Here, the drone dive (or emersion) is examined at constant speed: it is in steady-state gliding,
the geometry of total forces is explained in Figure 7a. Figure 7b shows the Eppler E838 characteristic
“Cl vs. Alpha (=angle of attack 𝛼௧௧)”. (a)
(b)
Figure 7. Dynamic balance of the forces (a) Drone geometrical balance of the forces; (b) Cl vs. Alpha
(=angle of attack 𝛼
) diagram for Eppler E838 aerofoil
Figure 7. Dynamic balance of the forces (a) Drone geometrical balance of the forces; (b) Cl vs. Alpha
(=angle of attack, αatt diagram for Eppler E838 aerofoil. (a) (b) (a) (b) Figure 7. Dynamic balance of the forces (a) Drone geometrical balance of the forces; (b) Cl vs. Alpha
( a
le of atta k
) dia
a
fo E
le E838 ae ofoil
Figure 7. Dynamic balance of the forces (a) Drone geometrical balance of the forces; (b) Cl vs. Alpha
(=angle of attack, αatt diagram for Eppler E838 aerofoil. (
g
,
௧௧)
g
pp
At equilibrium for the dynamics on the vertical plane at constant speed we have
At equilibrium, for the dynamics on the vertical plane at constant speed we have: n the vertical plane at constant speed we have:
𝑊்ை்
ሬሬሬሬሬሬሬሬሬሬ⃗+ 𝐿ሬ⃗+ 𝐷ሬሬ⃗= 0
(5)
𝐿= 1
2 𝜌𝑣ଶ𝑆𝐶
(6)
→
WTOT +
→
L +
→
D = 0
(5)
L = 1
2ρv2SCL
(6) on the vertical plane at constant speed we have:
𝑊்ை்
ሬሬሬሬሬሬሬሬሬሬ⃗+ 𝐿ሬ⃗+ 𝐷ሬሬ⃗= 0
(5)
→
WTOT +
→
L +
→
D = 0
(5) (5)
(5) The expression for the lift is:
The expression for the lift is: The expression for the lift is:
The expression for the lift is: 𝐿= 1
2 𝜌𝑣ଶ𝑆𝐶
(6)
L = 1
2ρv2SCL
(6) (6)
(6) 2
According to the Eppler E838 characteristic “Cl vs. Alpha” (Figure 7b) when the angle of attack
𝛼௧௧ = 0° (is null) the 𝐶 is zero so that the lift force 𝐿 is null. 2.2.2. Dynamic Balance on the Vertical Plane
2.2.2. Dynamic Balance on the Vertical Plane This shows that the drone cannot
progress horizontally at constant speed (straight and level): the only mission profile allowed is a
sawtooth curve [65]
According to the Eppler E838 characteristic “Cl vs. Alpha” (Figure 7b) when the angle of attack
αatt = 0◦(is null) the CL is zero so that the lift force L is null. This shows that the drone cannot progress
horizontally at constant speed (straight and level): the only mission profile allowed is a sawtooth
curve [65]. where:
where: WBA = Weight of the battery pack. 𝑊 = Weight of the battery pack. BA
Weight of the battery pack. OT = Weight of the oil tank.
g
y p
ை் = Weight of the oil tank. WOT = Weight of the oil tank. 𝑊ை் = Weight of the oil tank. WOT = Weight of the oil tank. 𝑊ை் = Weight of the oil tank. WOT = Weight of the oil tank. 𝑊ை் = Weight of the oil tank. g
WGB = Weight of the naked glider (without oil tank and batteries). For “dry weight”, we mean
the weight of the vehicle out of the sea, without the hydrostatic force. 𝑊ீ = Weight of the naked glider (without oil tank and batteries). For “dry weight”, we mean
the weight of the vehicle out of the sea, without the hydrostatic force. So Equation (2) becomes:
So Equation (2) becomes: So Equation (2) becomes:
So Equation (2) becomes: X
Fz = WBA + WOT + WGB + BGB + BBB = 0
(4)
𝐹௭= 𝑊+ 𝑊ை்+ 𝑊ீ+ 𝐵ீ+ 𝐵= 0
(4) (4)
(4) This series of equations will be useful later to establish the drone descent attitude. This series of equations will be useful later to establish the drone descent attitude. This series of equations will be useful later to establish the drone descent attitude. This series of equations will be useful later to establish the drone descent attitude. 2.2.1. Static Buoyancy
2.2.1. Static Buoyancy
2.2.1. Static Buoyancy y
y
Now, consider our drone (see Figure 6): it is in steady state. Now, consider our drone (see Figure 6): it is in steady state. Now, consider our drone (see Figure 6): it is in steady state. y
y
Now, consider our drone (see Figure 6): it is in steady state. Now, consider our drone (see Figure 6): it is in steady state. Now, consider our drone (see Figure 6): it is in steady state. Now, consider our drone (see Figure 6): it is in steady state. Figure 6. Drone in buoyancy Balance. Figure 6. Drone in buoyancy Balance. Figure 6. Drone in buoyancy Balance. Figure 6. Drone in buoyancy Balance. Figure 6. Drone in buoyancy Balance. Figure 6. Drone in buoyancy Balance. g
y
y
In these conditions the total weight WTOT of the glider is given by:
In these conditions, the total weight WTOT of the glider is given by:
In these conditions, the total weight WTOT of the glider is given by: In these conditions, the total weight WTOT of the glider is given by:
𝑊்ை்= −𝑊ௐ+ 𝐵ீ+ 𝐵
(2)
In these conditions, the total weight WTOT of the glider is given by:
𝑊்ை்= −𝑊ௐ+ 𝐵ீ+ 𝐵
(2)
h
In these conditions, the total weight WTOT of the glider is given by:
WTOT = −WDW + BGB + BBB
(2) (2)
(2)
(2) 𝑊்ை்= −𝑊ௐ+ 𝐵ீ+ 𝐵
𝑊்ை்= −𝑊ௐ+ 𝐵ீ+ 𝐵
WTOT = −WDW + BGB + BBB where:
where:
where: 𝑊்ை்= Net total “weight” in the water. 𝑊்ை் = Net total “weight” in the water. WTOT = Net total “weight” in the water. 𝑊்ை் Net total weight in the w
WDW = Dry Weight of the glider. BBB = Buoyancy of the oil bladder. BGB = Buoyancy of the naked glider. 6 of 17 Sensors 2019, 19, 1831
𝑊ௐ = Dry W
𝐵 = Buoyan The expression of dry weight is:
The expression of dry weight is: WDW = WBA + WOT + WGB
(3)
𝑊ௐ= 𝑊+ 𝑊ை்+ 𝑊ீ
(3) WDW = WBA + WOT + WGB
𝑊ௐ= 𝑊+ 𝑊ை்+ 𝑊ீ (3)
(3) 2 2 3 Glider Trajectory
2.2.3. Glider Trajectory 𝑐: is the mean aerodynamic cho
𝛼௧௧: is the angle of attack
c: is the mean aerodynamic chord. αatt: is the angle of attack. 𝑐: is the mean aerodyn
𝛼௧௧: is the angle of atta
αatt: is the angle of attack. ௧௧
g
For the other coefficients we ha
For the other coefficients, we have: ௧௧
g
For the other coefficients we ha
For the other coefficients, we have: e
ave:
𝐶(𝛼) = 𝐶
ఈ∙𝛼௧௧
(𝛼) = 𝐶
+ 𝐶
ఈ∙𝛼௧௧
ଶ
𝐶(𝛼) = 𝐶
ఈ∙𝛼௧௧
(8)
d
f
b
h f
h
CL(α) = Cα
L·αatt
CD(α) = C0
D + Cα
D·α2
att
Cm(α) = Cα
m·αatt
(8)
7 of 17 (8
(8) The coefficient of drag 𝐶 is composed of two members: the first 𝐶 is insensitive to the an
of attack and is constant; the second one (𝐶
ఈ∙𝛼௧௧
ଶ) is instead a function of the square of the ang
Note that the zero lift coefficient 𝐶
= 0 because the wing profile that has been chosen for our proj
is symmetrical (type Eppler 883). Now is necessary to separate the contributions of the fuselage (bod
and of the wings, for the three factors of lift, friction and pitching moment; so the expression is:
൝
𝕃= 𝐿+ 𝐿௪
The coefficient of drag CD is composed of two members: the first C0
D is insensitive to the angle of
attack and is constant; the second one (Cα
D·α2
att) is instead a function of the square of the angle. Note
that the zero lift coefficient C0
L = 0 because the wing profile that has been chosen for our project is
symmetrical (type Eppler 883). Now is necessary to separate the contributions of the fuselage (body)
and of the wings, for the three factors of lift, friction and pitching moment; so the expression is:
oth by the wings. Moreover, unlike gliders in air, AUVs can have ascending glide slopes if the
uoyancy is positive, producing a negative sink rate. The buoyancy engine of the glider allows changing its net buoyancy into alternating positive
negative states, thereby imparting it with the ability to string together a succession of descending
ascending glide slopes referred to as a sawtooth glide. 2 2 3 Glider Trajectory
2.2.3. Glider Trajectory This parameter is necessary to
d
fil [66]
k and is constant; the second one (𝐶
ఈ∙𝛼
h
l f
ff
b
rodynamic chord
cm is a non-dimensional coefficient. constant descent profile [66]. 2.2.4. Gliding Forces
that the zero lift coefficient 𝐶
= 0 because the wing profile that has been chosen for our project
mmetrical (type Eppler 883). Now is necessary to separate the contributions of the fuselage (body)
of the wings, for the three factors of lift, friction and pitching moment; so the expression is:
y
gle of attack
coefficients, we have:
𝐶(𝛼)
𝐶ఈ𝛼
This parameter is necessary to know the exact attitude and therefore the αatt to obtain a constant
descent profile [66]. constant descent profile [66]. 2 2 4 Gliding Forces
that the zero lift coefficient 𝐶
= 0 because the wing profile that has been chosen for our project
mmetrical (type Eppler 883). Now is necessary to separate the contributions of the fuselage (body)
f th
i
f
th th
f
t
f lift f i ti
d
it hi
t
th
i
i
y
gle of attack
coefficients, we have:
This parameter is necessary to know the exact attitude and therefore the αatt to obtain a constant
descent profile [66]. 2 2 3 Glider Trajectory
2.2.3. Glider Trajectory j
y
Because its motion is due to the difference between the forces of weight and buoyancy, the glider
is unable to proceed straight and level, thus being forced to follow a dive/climb trajectory made
Because its motion is due to the difference between the forces of weight and buoyancy, the glider
is unable to proceed straight and level, thus being forced to follow a dive/climb trajectory made smooth
by the wings. Moreover, unlike gliders in air, AUVs can have ascending glide slopes if the net buoyancy
is positive, producing a negative sink rate. Sensors 2019, 19, 1831
Sensors 2019, 19, 7 of 177 o The buoyancy engine of the glider allows changing its net buoyancy into alternating positive and
negative states, thereby imparting it with the ability to string together a succession of descending and
ascending glide slopes referred to as a sawtooth glide. smooth by the wings. Moreover, unlike gliders in air, AUVs can have ascending glide slopes if
net buoyancy is positive, producing a negative sink rate. The buoyancy engine of the glider allows changing its net buoyancy into alternating posit
and negative states, thereby imparting it with the ability to string together a succession of descend The behaviour of the vehicle is considered in case of a simple glide slope (refer to Figure 7a)
g
,
y
p
g
y
g
g
and ascending glide slopes referred to as a sawtooth glide. Lift = L = qSCL
Drag = D = qSCD
hing moment =
is considered in case of a simple glide slope (refer to Figure 7a):
𝐿𝑖𝑓𝑡= 𝕃= 𝑞𝑆𝐶
𝐷𝑟𝑎𝑔= 𝔻= 𝑞𝑆𝐶
𝑖𝑛𝑔 𝑚𝑜𝑚𝑒𝑛𝑡= 𝕞= 𝑞𝑆𝑐𝐶
(7
= qScCm
(7) Lift = L = qSCL
Drag = D = qSCD
s considered in case o
𝐿𝑖𝑓𝑡= 𝕃= 𝑞𝑆𝐶
𝐷𝑟𝑎𝑔= 𝔻= 𝑞𝑆𝐶 (7
(7) Pitching moment =
𝑃𝑖𝑡𝑐ℎ𝑖𝑛𝑔 𝑚𝑜𝑚𝑒𝑛𝑡= 𝕞= 𝑞𝑆𝑐𝐶
= qScCm whe
where: 𝑞=
ଵ
ଶ𝜌𝑣ଶ: is the dynamic pressure
𝑆: is the characteristic area
𝑐: is the mean aerodynamic chord
𝛼௧௧: is the angle of attack
q = 1
2ρv2: is the dynamic pressure. S: is the characteristic area. c: is the mean aerodynamic chord. αatt: is the angle of attack. F
h
h
ffii
h 𝑞=
ଵ
ଶ𝜌𝑣ଶ: is the dynamic pressu
𝑆: is the characteristic area
q = 1
2ρv2: is the dynamic pressure. S: is the characteristic area. 2 2 3 Glider Trajectory
2.2.3. Glider Trajectory ൝𝔻= 𝐷+ 𝐷
𝕞= 𝑚+ 𝑚௪
(9
es (approximated) equations and the simplifications above cit
ecomes:
L = Lb + Lw
D = Db + Dw
d in case of a simple glide slope (refer to Figure 7a):
= 𝕃= 𝑞𝑆𝐶
= 𝔻= 𝑞𝑆𝐶
𝑡= 𝕞= 𝑞𝑆𝑐𝐶
(7)
= mb + mw
(9)
7 of 17
Vs can have ascending glide slopes if the ve cit
(9) ⎪⎪
⎧
𝕃= 𝑞ඥ𝑉ଷ൜𝐶
ഀ∙𝛼௧௧+ 𝑆௪
√𝑉ଷ∙𝐶
௪ഀ∙𝛼௧௧ൠ
ඥ
൜ൣ
൧
𝑆௪ൣ
൧ൠ
re:
𝑞=
ଵ
ଶ𝜌𝑣ଶ: is the dynamic pressure
According to the Navier-Stokes (approximated) equations and the simplifications above cited, the
previous system of equations becomes:
ositive, producing a negative sink rate. cy engine of the glider allows changing its net buoyancy into alternating positive
es, thereby imparting it with the ability to string together a succession of descending 𝑞ඥ𝑉൜ൣ𝐶+ 𝐶
𝛼௧௧൧+
√𝑉ଷൣ𝐶
+ 𝐶
𝛼௧௧൧ൠ
= 𝑞ඥ𝑉ଷቊ𝐶
ഀ∙𝛼௧௧−𝑐
𝑙/ೢ∙𝑆௪
𝑐∙√𝑉ଷ
∙𝐶
௪ഀ∙𝛼௧௧ቋ
(10
wing area
area
namic chord
tack
ents, we have:
𝐶(𝛼) = 𝐶
ఈ∙𝛼௧௧
ଶ
L = q
√
V3
Cbα
L ·αatt +
Sw
√
V3 ·Cwα
L ·αatt
D = q
√
V3
h
Cb0
D + Cbα
D ·α2
att
i
+
Sw
√
V3
h
Cw0
D + Cwα
D ·α2
att
i
s a sawtooth glide. sidered in case of a simple glide slope (refer to Figure 7a):
𝐿𝑖𝑓𝑡= 𝕃= 𝑞𝑆𝐶
𝑟𝑎𝑔= 𝔻= 𝑞𝑆𝐶
𝑚𝑜𝑚𝑒𝑛𝑡= 𝕞= 𝑞𝑆𝑐𝐶
(7)
= q
√
V3
Cbα
m ·αatt −cm
lcb/acw·Sw
c·
√
V3 ·Cwα
L ·αatt
(10) (10) where: 𝑙/ೢ is the distance between the mean aerodynamic and the center of buoyancy. 𝑐is a non-dimensional coefficient
𝐶(𝛼) = 𝐶
ఈ∙𝛼௧௧
he dynamic pressure
Sw is the wing area. 𝑙/ೢ is the dis
𝑐is a non-dim
e dynamic pressure
Sw is the wing area. 𝑐 is a non-dimensional coefficient
Thi
t
i
t
k
th
t
ttit d
d th
f
th
e coefficient of drag 𝐶 is composed of two members: the first 𝐶
is insensitive to the a
k
d i
t
t th
d
(𝐶ఈ
ଶ) i i
t
d
f
ti
f th
f th
eristic area
w
g
lcb/acw is the distance between the mean aerodynamic and the center of buoyancy. In order to a
g ,
𝐶(𝛼) =
𝐶(𝛼) = 𝐶
2.2.4. Gliding Forces Restoring Moment on the Vertical Plan In order to a
g ,
𝐶(𝛼) =
𝐶(𝛼) = 𝐶
2.2.4. Gliding Forces therefore the balance is not null: from the buoyant force expression, the change in volume needed
the buoyancy engine for a full dive is calculated as:
𝐹௨ௗ= ∆𝐵≠0
(11
൝𝔻= 𝐷+ 𝐷௪
𝕞= 𝑚+ 𝑚௪
(9)
According to the Navier-Stokes (approximated) equations and the simplifications above cited,
revious system of equations becomes:
𝐶(𝛼)
𝐶+ 𝐶
𝛼௧௧
𝐶(𝛼) = 𝐶
ఈ∙𝛼௧௧
(8)
nt of drag 𝐶 is composed of two members: the first 𝐶
is insensitive to the angle
nstant; the second one (𝐶
ఈ∙𝛼௧௧
ଶ) is instead a function of the square of the angle. In order to allow the vehicle to glide, it is necessary to create a differential buoyancy force and
therefore the balance is not null: from the buoyant force expression, the change in volume needed for
the buoyancy engine for a full dive is calculated as: ancy force due to the difference of volume of the bladder:
ௗ= ∆𝐵= 1
2 𝜌𝑔∙∆𝑉ௗௗ
(12
ଷ∙𝐶
௪ഀ∙𝛼௧௧ൠ
𝑆
ng profile that has been chosen for our project
parate the contributions of the fuselage (body)
d pitching moment; so the expression is:
X
FZumbalanced = ∆BBB , 0
(11) (11) ancy force due to the difference of volume o
= ∆𝐵
= 1 𝜌𝑔∙∆𝑉
∙𝐶
௪ഀ∙𝛼௧௧ൠ
𝑆
ng profile that has been chosen for our proje
arate the contributions of the fuselage (bod
d
h
h
X
FZumbalanced = ∆BBB , 0 ncy force due to the differenc
= ∆𝐵
= 1 𝜌𝑔∆𝑉
∙𝐶
௪ഀ∙𝛼௧௧ൠ
𝑆
ng profile that has been chosen
arate the contributions of the
h
h
X
FZumbalanced = ∆BBB , 0 8 of 17 Sensors 2019, 19, 1831 In which ∆BBB , 0 is the buoyancy force due to the difference of volume of the bladder: In which ∆BBB , 0 is the buoyancy force due to the difference of volume of the bladder: X
FZumbalanced = ∆BBB = 1
2 ρg·∆Vbladder
(12) (12) where:
ρ = Seawater density (average 1.025 kg/l). ρ = Seawater density (average 1.025 kg/l). ρ = Seawater density (average 1.025 kg/l). g = Gravity approx. to 9.81 m/s2. g = Gravity approx. to 9.81 m/s2. g
y pp
∆Vbladder = Volume difference of the bladder. The relationship between difference volume of the bladder and buoyancy force is: ∆Vbladder =
2
ρg ·∆BBB
(13) ∆Vbladder =
2
ρg ·∆BBB (13) 2.2.5. Restoring Moment on the Vertical Plan 2.2.5. 2.2.6. Uncertainty in the Depth Control In this part, the problem of uncertainty in controlling the depth of the drone is examined. In the
case where the drone has a very low vertical speed, due to the high viscosity of the water, the vehicle
stops (and therefore stabilizes itself in the vertical plan) in a few centimeters of water, so this is not
a problem. Consider the case in which the drone is gliding, at a constant speed (Vz < 0): at a depth of
+5 m compared to the “target depth”, the bladder swells up to make the drone assume a neutral
buoyancy. From a dynamic point of view, our simulations show us that the drone will behave like a
mass-spring-damper system, whose temporal behavior is described in Equation (23) and visible in
Figure 8 (blue line). Figure 8 (blue line).
zCG(t) = Ze−ζωnt· cos
p
1 −ζ2ωnt −Φ
ωn = 2π fn
(23) (23) The maximum precision in the measurement of depth is not given by the precision of the
instrument, in this case a piezoelectric depth meter can be accurate up to 2.5 cm, but in the architecture
of the drone. Because of all the possible attitudes of the vehicle, it is not known if the transducer is
placed higher or lower than the centre of gravity: so, unless more than one transducer is installed
(which is impractical) the only consideration is that the maximum distance at which the transducer
should be placed from the centre of gravity is precisely 49.7 cm, so, the maximum range of precision
obtainable from the instrument is: (24) δz = 0.497 m
(24) As shown in Figure 8 (blue line) the natural dynamic stability of the vehicle, i.e., the oscillation
within the error band, is obtained after a time Tnormal = 9.78 s which is not acceptable (see Equation (25)). It is not acceptable for two reasons: first of all because of the long damping time of the oscillation;
secondly for the overshoot of the “target depth”: the depth limit may have been placed not only by the
type of mission but also by the nature (orography) of the seabed. 2.2.5. Restoring Moment on the Vertical Plan The position of the center of mass is given by: The position of the center of mass is given by: The position of the center of mass is given by: rCG =
R
r·ρ(r)dV
R
ρ(r)dV
(14) rCG =
R
r·ρ(r)dV
R
ρ(r)dV (14) where: ρ( )
y
dV: is the considered volume. dV: is the considered volume. i th di t
id
d f
th
f
f r: is the distance considered from the reference frame. r: is the distance considered from the reference frame. The gliders masses define above, is: The gliders masses define above, is: The gliders masses define above, is: rCG =
P
i ri·Wi
P
i Wi
= rBA·WBA + rOT·WOT + rGB·WGB
WBA + WOT + WGB
(15) (15) The vectorial balance of the forces becomes: The vectorial balance of the forces becomes:
Fgravitational = WDW
RTz
Fbuoyancy = −VDispalacement·g
RTz
(16) (16) where VDispalacement is the volume of the drone (submerged). VDispalacement is the volume of the drone (submerged). VDispalacement is the volume of the drone (submerged). T p
RT is the rotation function around the reference frame. p
RT is the rotation function around the reference frame. z is the upward direction. 2.2.5. Restoring Moment on the Vertical Plan p
hen the geometrical centre of the body is offset from the CG frame, the resulting torque TG is
y: When the geometrical centre of the body is offset from the CG frame, the resulting torque TG is
given by:
T TG = rCG × WDW
RTz
(17) (17) G
CG
DW
(
)
The expression of rCG is:
rCG =
xCG
yCG
zCG
(18)
The expression of ˆrCG is:
0
The expression of rCG is: rCG =
xCG
yCG
zCG
(18) (18) The expression of ˆrCG is: ˆrCG =
0
−zCG
yCG
zCG
0
−xCG
−yCG
xCG
0
(19) (19) Therefore, we have: Therefore, we have:
TG = WDW·ˆrCG
RTz
(20) TG = WDW·ˆrCG
RTz
(20) (20) Sensors 2019, 19, 1831 9 of 17 According to which, when the body frame is coincident with the centre of the figure: According to which, when the body frame is coincident with the centre of the figure: WDW·rCG = rGB·BGB + rBB·BBB
(21) (21) Moreover, the resulting balancing torque is TG = (rGB·BGB + rBB·BBB)·g
RTz
(22) (22) This parameter is necessary to know the exact torque and therefore the position to which the
servomechanism to move the battery pack will have to obtain a constant descent profile [67]. This parameter is necessary to know the exact torque and therefore the position to which the
servomechanism to move the battery pack will have to obtain a constant descent profile [67]. 2.2.6. Uncertainty in the Depth Control 2.2.6. Uncertainty in the Depth Control Model parameters. fn
ζnormal
ζcompensated
Z
1
0.0794
0.15779
4.972 2.2.6. Uncertainty in the Depth Control Therefore, such behavior could lead
to a collision of the vehicle with the seabed itself or a known obstacle, thus leading to damage: (
zCG(t)normal ≤δz
Tnormal = 9.78 s
(25) (25) The solution is to use the bladder as a “hydrostatic parachute” that changes the damping conditions
of the system. In this case the simulation have evidenced that, if you place the bladder at maximum
buoyancy (Figure 8—red line) shortly after reaching “+5” depth, the resulting behavior of the vehicle
is visible in Figure 8 (azure line). In this case the tolerance band is reached in Tcompensated = 4.85 s (see Equation (26)), half the
previous time and there is no danger of overshooting the depth, thus keeping us always in safe
conditions: ( zCG(t)compensated ≤δz
Tcompensated = 4.85 s
(26) ( zCG(t)compensated ≤δz
Tcompensated = 4.85 s (26) 10 of 17 10 of 17 Sensors 2019, 19, 1831 From a mathematical point of view, the system changes the damping factor, increasing it
considerably in fact, ζnormal < ζcompensated. The model parameters are shown in Table 1 below. Sensors 2019, 19, x
10 of 17 From a mathematical point of view, the system changes the damping factor, increasing it
considerably in fact, ζnormal < ζcompensated. The model parameters are shown in Table 1 below. Sensors 2019, 19, x
10 of 17 Figure 8. Drone depth dynamic behavior: in the blue line the normal (uncompensated) damping; the
azure line the compensated behavior; dotted lines are the upper and lower limit. The red line (out of
scale) is the bladder’s “hydrostatic chute” action. Figure 8. Drone depth dynamic behavior: in the blue line the normal (uncompensated) damping; the
azure line the compensated behavior; dotted lines are the upper and lower limit. The red line (out of
scale) is the bladder’s “hydrostatic chute” action. Figure 8. Drone depth dynamic behavior: in the blue line the normal (uncompensated) damping; the
azure line the compensated behavior; dotted lines are the upper and lower limit. The red line (out of
scale) is the bladder’s “hydrostatic chute” action. Figure 8. Drone depth dynamic behavior: in the blue line the normal (uncompensated) damping; the
azure line the compensated behavior; dotted lines are the upper and lower limit. The red line (out of
scale) is the bladder’s “hydrostatic chute” action. mulate the behavior of the Eppler 838 aerofoil an
pensive but more sophisticated 3D simulation
Table 1. purpose, the program “Java
2.2.7. Dynamic Simulation several traditional methods for aerofoil analysis. The backbone of the program consists of two
methods:
•
The evaluation of the potential flow. The analysis is done with a higher order panel method
(linear varying vorticity distribution) and it calculates the local, inviscid flow velocity along the
surface of the aerofoil for any desired angle of attack. In the section, we simulate the behavior of the Eppler 838 aerofoil and the fuselage to evaluate,
before a detailed and expensive but more sophisticated 3D simulation, if the proportions and dimensions
of the drone fall within the range of measures evaluated as a requirement. For this purpose, the program
“JavaFoil—Analysis of Airfoils” [68] is used. JavaFoil is a program which uses several traditional
methods for aerofoil analysis. The backbone of the program consists of two methods: •
The evaluation of the boundary layer. The analysis is steps along the upper and the lower
surfaces of the aerofoil, starting at the stagnation point, solves a set of differential equations to
find the various boundary layer parameters, according to the integral method. •
The evaluation of the potential flow. The analysis is done with a higher order panel method (linear
varying vorticity distribution) and it calculates the local, inviscid flow velocity along the surface
of the aerofoil for any desired angle of attack. The equations and criteria for transition and separation are based on the procedures described
by Eppler. A standard compressibility correction according to Karman and Tsien has been
implemented to take moderate Mach number effects into account. Usually this means Mach numbers
•
The evaluation of the boundary layer. The analysis is steps along the upper and the lower surfaces
of the aerofoil, starting at the stagnation point, solves a set of differential equations to find the
various boundary layer parameters, according to the integral method. between zero and 0.5 [69–72]. The simulation shows the critical parameters for the hydrodynamic behaviour of the model: the
results are given in Table 2 below [73–78]. Table 2. Body and wing critical parameters [Speed = 0.52 kts, Re = 104 and 𝛼௧௧ = 3.3°]. The equations and criteria for transition and separation are based on the procedures described by
Eppler. A standard compressibility correction according to Karman and Tsien has been implemented
to take moderate Mach number effects into account. Usually this means Mach numbers between zero
and 0.5 [69–72]. purpose, the program “Java
2.2.7. Dynamic Simulation ters for the hydrodynam
ters [Speed = 0.52 kts, Re =
w0
D
Cbα
D
Cwα
D
676
0.00018
0.001515
3.3◦around the body
express the induced resi
ttack is provided in Figu
= 3.3° around the body 11 of 17 Sensors 2019, 19, 1831 The simulation shows the critical parameters for the hydrodynamic behaviour of the model: the
results are given in Table 2 below [73–78]. The trend of the flow lines with αatt = 3.3◦around the body and the wing are visible in
Figure 9a,b, below. Sensors 2019, 19, x
11 of 17 For completeness, in order to be able to express the induced resistance and the stall delay, the
behavior of the aerofoil at different angles of attack is provided in Figure 10. The trend of the flow lines with 𝛼௧௧ = 3.3° around the body and the wing are visible in
Figure 9a,b, below. (a)
(b)
Figure 9. Flow field and pressure gradient of conditions: Speed = 0.52 kts, Re = 104 and
𝛼௧௧= 3.3°, (a) Body profile of the drone; (b) Eppler 838 Aerofoil. Figure 9. Flow field and pressure gradient of conditions: Speed = 0.52 kts, Re = 104 and αatt = 3.3◦,
(a) Body profile of the drone; (b) Eppler 838 Aerofoil. (b) Figure 9. Flow field and pressure gradient of conditions: Speed = 0.52 kts, Re = 104 and
𝛼
3 3° (a) Body profile of the drone; (b) Eppler 838 Aerofoil
Figure 9. Flow field and pressure gradient of conditions: Speed = 0.52 kts, Re = 104 and αatt = 3.3◦,
(a) Body profile of the drone; (b) Eppler 838 Aerofoil. 12 of 17
12 of 17 Sensors 2019, 19, 1831
Sensors 2019, 19, x 19, 19, 1831
19, 19, x
(a)
(b)
(c)
ure 10. Flow field and pressure gradient of conditions (Speed = 0.52 kts, Re = 104) are shown at
owing 𝛼௧௧ : (a) 0.5° (b) 4.0° (c) 15.0°. ure 10. Flow field and pressure gradient of conditions (Speed = 0.52 kts, Re = 104) are shown at
owing αatt: (a) 0.5◦(b) 4.0◦(c) 15.0◦. (b) (c) (c) Figure 10. Flow field and pressure gradient of conditions (Speed = 0.52 kts, Re = 104) are shown at the
following 𝛼
: (a) 0 5° (b) 4 0° (c) 15 0°
Figure 10. 3. Conclusions This paper reports part of several preliminary studies of the Underwater Drones Group of
the Università degli Studi “Roma Tre” Science Department and follows the route traced on several
conference papers presented at the IEEE International Workshop on Metrology for the Sea (MetroSea);
this part is dedicated to the design and engineering study of the part relating to the UAV buoyancy. This paper highlights the large series of considerations and structural dimensions, going down in
great detail, of the Underwater Glider Mk. III that is currently in an advanced development phase. The real novelties of this work are highlighted in the development due to two strong constraints that
our group inserted during the design of this AUV: the first is to evaluate the project always under the
most conservative (pessimistic) operating conditions; the second is to evaluate how any changes made
to the subsystem in development (in our case the buoyancy s/s) is reflected (and forced) on all the other
parts (or subsystems). All the results both from the partial simulations and from the construction and
testing of subsystems will then be used in the operating vehicle. The first section is dedicated to the design and engineering study of the part relating to the UAV
buoyancy. In the first section, the architecture, the internal arrangement of the sub glider, the type of
mission profile and the maximum requirements for the performances are broadly described. In the
second part, the buoyancy system is described from an engineering-construction point of view: the
solutions developed and implemented in a working prototype were illustrated. The last part describes the mathematical requirements for sizing the vehicle. Firstly, the static
requirements that are used to determine the mechanical and dimensional sizing of the buoyancy
engine are examined. Then the dynamic stability (on the vertical plane) of the vehicle is analysed: this
quantifies the forces involved during the “glide”. The trajectory is analysed to decide the attitude and
the angle of attack: the latter is necessary, in stationary conditions, to determine the work point of the
profile or the position of the profile in the diagram “Cl vs. Alpha”. purpose, the program “Java
2.2.7. Dynamic Simulation Flow field and pressure gradient of conditions (Speed = 0.52 kts, Re = 104) are shown at the
following αatt: (a) 0.5◦(b) 4.0◦(c) 15.0◦. Sensors 2019, 19, 1831 13 of 17 13 of 17 3. Conclusions In order to have a constant attitude,
it is necessary to balance the moments in the vertical plane so that, once the wing profile is “started”
and in progressive acceleration (i.e., while is nearly to reach the terminal velocity) the pitch up effects
must be compensated by the movement of the centre of gravity. An evaluation of the uncertainty in
the depth control is also provided. Lastly, a simplified simulation is introduced in order to observe the hydrodynamic behaviour of
the fuselage (limited to the profile) and of the aerofoil at different angles of attack, to highlight the
stall characteristics. Author Contributions: The authors have made an equal contribution in all the sections of this article. Funding: This research received no external funding. Conflicts of Interest: The authors declare no conflict of interest. References 1. Wood, S. Autonomous underwater gliders. In Underwater Vehicles; Chapter 26; Inzartsev, A.V., Ed.; In-Tech:
Vienna, Austria, 2009; pp. 499–524. 1. Wood, S. Autonomous underwater gliders. In Underwater Vehicles; Chapter 26; Inzartsev, A.V., Ed.; In-Tech:
Vienna, Austria, 2009; pp. 499–524. 2. Kayton, M.; Fried, W.R. Avionics Navigation Systems; Wiley: New York, NY, USA, 2007; ISBN 9780471547952. 3. Jenkins, S.A.; Humphreys, D.E.; Sherman, J.; Osse, J.; Jones, C.; Leonard, N.; Graver, J.; Bachmayer, R.;
Clem, T.; Carroll, P.; et al. Underwater Glider System Study; Technical Report No. 53; Scripps Institution of
Oceanography: San Diego, CA, USA, May 2003. g
p y
g
y
4. Graver, J.G. Underwater Gliders: Dynamics, Control and Design. Ph.D. Thesis, Princeton University,
Princeton, NJ, USA, 2005. 5. Kan, L.; Zhang, Y.; Fan, H.; Yang, W.; Chen, Z. MATLAB-Based simulation of buoyancy-driven underwater
glider motion. J. Ocean Univ. Chin. 2007, 7, 113–118. [CrossRef] 6. Woolsey, C.A.; Leonard, N.E. Stabilizing underwater vehicle motion using internal rotors. Automatica 2002,
38, 2053. [CrossRef] 6. Woolsey, C.A.; Leonard, N.E. Stabilizing underwater vehicle motion using internal rotors. Automatica 2002,
38, 2053. [CrossRef] Sensors 2019, 19, 1831 14 of 17 7. Jekeli, C. Inertial Navigation Systems with Geodetic Applications; Walter de Gruyter Berlin: New York, NY, USA,
2001. 8. Smith, R.N.; Chao, Y.; Jones, B.H.; Caron, D.A.; Li, P.; Sukhatme, G.S. Trajectory Design for Autonomous
Underwater Vehicles based on Ocean Model Predictions for Feature Tracking. In Field and Service Robotics;
Springer: Berlin/Heidelberg, Germany, 2010; pp. 263–273, 2009. 9. Mitchell, B.; Wilkening, E.; Mahmoudian, N. Low cost underwater gliders for littoral marine research. In Proceedings of the American Control Conference (ACC), Seattle, WA, USA, 17–19 June 2013; pp. 1412–1417. g
g
In Proceedings of the American Control Conference (ACC), Seattle, WA, USA, 17–19 June 2013; pp. 1412–1417. 10. Meyer, D. Glider Technology for Ocean Observations: A Review. Ocean Sci. Discuss. 2016, 2016, 1–26. [CrossRef] 10. Meyer, D. Glider Technology for Ocean Observations: A Review. Ocean Sci. Discuss. 2016, 2016, 1–26. [CrossRef] 11. Davis, R.E.; Eriksen, C.C.; Jones, C.P. Autonomous buoyancy-driven underwater gliders. In Technology
and Applications of Autonomous Underwater Vehicles; Griffiths, G., Ed.; Taylor & Francis: London, UK, 2002;
pp. 37–58. 12. Wang, C.; Zhang, Z.; Gu, J.; Liu, J.; Miao, T. Design and Hydrodynamic Performance Analysis of Underwater
Glider Model. In Proceedings of the 2012 International Conference on Computer Distributed Control and
Intelligent Environmental Monitoring (CDCIEM), Hunan, China, 5–6 March 2012; pp. 225–227. References [CrossRef] 13. De Francesco, E.; De Francesco, E.; De Francesco, R.; Leccese, F.; Cagnetti, M. A proposal to update LSA
databases for an operational availability based on autonomic logistic. In Proceedings of the 2nd IEEE
International Workshop on Metrology for Aerospace, MetroAeroSpace, Benevento, Italy, 4–5 June 2015;
pp. 38–43. [CrossRef] 14. Eriksen, C.C.; Osse, T.J.; Light, R.D.; Wen, T.; Lehman, T.W.; Sabin, P.L.; Ballard, J.W.; Chiodi, A.M. Seaglider:
A longrange autonomous underwater vehicle for oceanographic research. IEEE J Oceanic Eng. 2001, 26,
424–436. [CrossRef] 15. Rudnick, D.L.; Eriksen, C.C.; Fratantoni, D.M.; Perry, M.J. Underwater Gliders for Ocean Research. Mar. Technol. Soc. J. 2004, 38, 1. 16. Austin, R. Unmanned Aircraft Systems; Wiley: Hoboken, NJ, USA, 2010. 17. Graver, J.G.; Liu, J.; Woolsey, C.; Leonard, N.E. Design and Analysis of an Underwater Vehicle for Controlled
Gliding. In Proceedings of the Conference on Information Sciences and Systems (CISS), Princeton, NJ, USA,
2–5 July 1998. 18. Leccese, F.; Cagnetti, M.; Giarnetti, S.; Petritoli, E.; Luisetto, I.; Tuti, S.; Ðurovi´c-Pejˇcev, R.; Ðorđevi´c, T.;
Tomaševi´c, A.; Bursi´c, V.; et al. A Simple Takagi-Sugeno Fuzzy Modelling Case Study for an Underwater
Glider Control System. In Proceedings of the 2018 IEEE International Workshop on Metrology for the
Sea; Learning to Measure Sea Health Parameters (MetroSea), Bari, Italy, 8–10 October 2018; pp. 262–267. [CrossRef] 19. Teledyne Webb Research. Slocum Electric Glider. Available online: http://www.webbresearch.com/ (accessed
on 1 February 2019). 20. Wood, S. State of Technology in Autonomous Underwater Gliders. Mar. Technol. Soc. J. 2013, 47, 5. [CrossRef] 20. Wood, S. State of Technology in Autonomous Underwater Gliders. Mar. Technol. Soc. J. 2013, 47, 5. [CrossRef]
21. Waldmann, C.; Kausche, A.; Iversen, M. MOTH-An underwater glider design study carried out as part of
th HGF lli
ROBEX I
P
di
f th 2014 IEEE/OES A t
U d
t
V hi l
(AUV) ,
gy
J
,
,
[
]
21. Waldmann, C.; Kausche, A.; Iversen, M. MOTH-An underwater glider design study carried out as part of
the HGF alliance ROBEX. In Proceedings of the 2014 IEEE/OES Autonomous Underwater Vehicles (AUV),
Oxford, UK, 25–29 July 2014; pp. 1–3. [CrossRef] 22. Davis, R.E.; Webb, D.C.; Regier, A.; Dufour, J. The Autonomous Lagrangian Circulation Explorer (ALACE). J. Atmos. Oceanic Technol. 1992, 9, 264–285. [CrossRef] 23. Seo, D.C.; Gyungnam, J.; Choi, H.S. Pitching control simulation of an underwater glider using CFD analysis. References In Proceedings of the Oceans—MTS/IEEE Kobe Techno-Ocean, Kobe, Japan, 8–11 April 2008; pp. 1–5. 24. Evans, C.D.; Riggins, R. The Design and Analysis of Integrated Navigation Systems Using Real INS and GPS
Data. In Proceedings of the IEEE 1995 National Aerospace and Electronics Conference, Dayton, OH, USA,
22–26 May 1995. 25. Yu, J.; Zhang, F.; Zhang, A.; Jin, W.; Tian, Y. Motion Parameter Optimization and Sensor Scheduling for the
Sea-Wing Underwater Glider. IEEE J. Oceanic Eng. 2013, 38, 243–254. [CrossRef] 26. Hussain, N.A.A.; Arshad, M.R.; Mohd-Mokhtar, R. Modeling and Identification of an Underwater Glider. In Proceedings of the 2010 International Symposium on Robotics and Intelligent Sensors (IRIS2010), Nagoya,
Japan, 8–11 March 2010. 27. Bohenek, B.J. The Enhanced Performance of an Integrate Navigation System. In A Highly Dynamic Environment;
Air Force Institute of Technology: Dayton, OH, USA, 1994. Sensors 2019, 19, 1831 15 of 17 28. Beard, R.W.; McLain, T.W. Small Unmanned Aircraft—Theory and Practice; Princeton University Press: Princeton,
NJ, USA, 2012. 28. Beard, R.W.; McLain, T.W. Small Unmanned Aircraft—Theory and Practice; Princeton University Press: Princeton,
NJ, USA, 2012. 29. Petritoli, E.; Leccese, F.; Ciani, L. Reliability assessment of UAV systems. In Proceedings of the 4th IEEE
International Workshop on Metrology for AeroSpace (MetroAeroSpace), Padua, Italy, 21–23 June 2017;
pp. 266–270. pp
30. Watts, A.C.; Ambrosia, V.G.; Hinkley, E.A. Unmanned Aircraft Systems in Remote Sensing and Scientific
Research: Classification and Considerations of Use. Remote Sens. 2012, 4, 1671–1692. [CrossRef] 31. Petritoli, E.; Leccese, F. Improvement of altitude precision in indoor and urban canyon navigation for
small flying vehicles. In Proceedings of the 2nd IEEE International Workshop on Metrology for Aerospace
(MetroAeroSpace 2015), Benevento, Italy, 4–5 June 2015; pp. 56–60. [CrossRef] 32. Andrade-Bustos, I.; Salgado-Jiménez, T.; García-Valdovinos, L.G.; Bandala-Sánchez, M. Stable Sliding PD
Control for underwater gliders: Experimental results. In Proceedings of the Oceans 2016 MTS/IEEE Monterey,
Monterey, CA, USA, 19–23 September 2016; pp. 1–7. 33. Pereira, A.; Heidarsson, H.; Caron, D.A.; Jones, B.H.; Sukhatme, G.S. A communication framework for
the cost-effective operation of slocum gliders in coastal regions. In Proceedings of the 7th International
Conference on Field and Service Robotics, Cambridge, MA, USA, 14–16 July 2009; pp. 1–10. 34. Skibski, C.E. Design of an Autonomous Underwater Glider focusing on External Wing Control Surfa
Sensor Integration. Bachelor’s Thesis, Florida Institute of Technology, Melbourne, FL, USA, 2009. 35. Isa, K.; Arshad, M.R. References An analysis of homeostatic motion control system for a hybrid-driven underwater glider. In Proceedings of the 2013 IEEE/ASME International Conference on Advanced Intelligent Mechatronics
(AIM), Wollongong, Australia, 9–12 July 2013; pp. 1570–1575. 36. Küchemann, D. The Aerodynamic Design of Aircraft; Pergamon international library of science, technology,
engineering, and social studie; Pergamon Press: Oxford, UK, 1978. 37. Parthasarathy, G.; Sree, D.S.; Manasa, B.L. Design Mathematical Modeling and Analysis of Underwater
Glider. Int. J. Sci. Res. 2015, 4, 711–714. 38. Song, D.L.; Yao, L.L.; Wang, Z.Y.; Han, L. Pitching Angle Control Method of Underwater Glider Based on
Motion Compensation. In Proceedings of the 2015 International Conference on Computational Intelligence
and Communication Networks (CICN), Jabalpur, India, 12–14 December 2015; pp. 1548–1551. 39. Smith, R.N.; Schwager, M.; Smith, S.L.; Rus, D.; Sukhatme, G.S. Persistent Ocean Monitoring with Underwater
Gliders Towards Accurate Reconstruction of Dynamic Ocean Processes. In Proceedings of the 2011 IEEE
International Conference Robotics and Automation (ICRA), Shanghai, China, 9–13 May 2011; pp. 1517–1524. 40. Techy, L.; Tomokiyo, R.; Quenzer, J.; Beauchamp, T.; Morgansen, K. Full Scale Wind Tunnel Study of the Seaglider
Underwater Glider; UWAA Technical Report UWAATR-2010-0002; University of Washington: Seattle, WA,
USA, September 2010. 41. Wilcox, J.S.; Bellingham, J.G.; Zhang, Y.; Baggeroer, A.B. Performance metrics for oceanographic surveys
with autonomous underwater vehicles. IEEE J Oceanic Eng. 2001, 26, 711–725. [CrossRef] 42. Petritoli, E.; Giagnacovo, T.; Leccese, F. Lightweight GNSS/IRS integrated navigation system for UAV vehicles. In Proceedings of the 2014 IEEE International Workshop on Metrology for Aerospace (MetroAeroSpace),
Benevento Italy, 29–30 June 2014; pp. 56–61. [CrossRef] In Proceedings of the 2014 IEEE International Workshop on Metrology for Aerospace (MetroAeroSpace),
Benevento Italy, 29–30 June 2014; pp. 56–61. [CrossRef] y
pp
43. Webb, D.C.; Simonetti, P.J.; Jones, C.P. SLOCUM: An underwater glider propelled by environmental energy. IEEE J Oceanic Eng. 2001, 26, 447–452. [CrossRef] 44. Stommel, H. The Slocum Mission. Oceanography 1989, 2, 22–25. [CrossRef] 45. Sherman, J.; Davis, R.E.; Owens, W.B.; Valdes, J. The autonomous underwater glider “Spray”. IEEE J Oceanic
Eng. 2001, 26, 437–446. [CrossRef] 46. Titterton, D.; Weston, J. Strapdown Inertial Navigation Technology, 2nd ed.; The Institution of Electrical Engineers
and The American Institute of Aeronautics and Astronautics: Reston, VA, USA, 2004. 47. Jun, B.H.; Park, J.Y.; Lee, F.Y.; Lee, P.M.; Lee, C.M.; Kim, K.; Lim, Y.K.; Oh, J.H. Development of the AUV
‘ISiMI’ and free running test in an ocean engineering basin. Ocean Eng. 2009, 36, 1. References In Proceedings of
the IMEKO TC19 Workshop on Metrology for the Sea, MetroSea 2017: Learning to Measure Sea Health
Parameters, Naples, Italy, 11–13 October 2017; pp. 7–12. 57. DSIAC. Reliability of UAVs and Drones. Available online: https://www.dsiac.org/resources/journals/dsiac/
spring-2017-volume-4-number-2/reliability-uavs-and-drones (accessed on 28 February 2019). 58. Etkin, B. Dynamic of Flight; John Wiley and Sons: Hoboken, NJ, USA, 1959. 58. Etkin, B. Dynamic of Flight; John Wiley and Sons: Hoboken, NJ, USA, 1959. 59. Tan, X.; Kim, D.; Usher, N.; Laboy, D.; Jackson, J.; Kapetanovic, A.; Rapai, J.; Sabadus, B.; Zhou, X. An Autonomous Robotic Fish for Mobile Sensing. In Proceedings of the 2006 IEEE/RSJ International
Conference on Intelligent Robots and Systems, Beijing, China, 9–15 October 2006; pp. 5424–5429. 60. Smith, R.N.; Pereira, A.; Chao, Y.; Li, P.P.; Caron, D.A.; Jones, B.H.; Sukhatme, G.S. Autonomous Underwater
Vehicle trajectory design coupled with predictive ocean models: A case study. In Proceedings of the 2010
IEEE International Conference on Robotics and Automation (ICRA), Anchorage, AK, USA, 4–8 May 2010;
pp. 4770–4777. 61. Petritoli, E.; Leccese, F.; Cagnetti, M. A High Accuracy Buoyancy System Control for an Underwater Glider. In Proceedings of the 2018 IEEE International Workshop on Metrology for the Sea; Learning to Measure Sea
Health Parameters (MetroSea), Bari, Italy, 8–10 October 2018; pp. 257–261. [CrossRef] 62. Techy, L.; Kristi Morganseny, A.; Woolseyz, C.A. Long-baseline acoustic localization of the Seaglider
underwater glider. In Proceedings of the American Control Conference (ACC) 2011, San Francisco, CA, USA,
29 June–1 July 2011; pp. 3990–3995. 63. Hussain, N.A.A.; Ali, S.S.A.; Saad, M.N.M.; Nordin, N. Underactuated nonlinear adaptive control approach
using U-model for multivariable underwater glider control parameters. In Proceedings of the 2016 IEEE
International Conference on Underwater System Technology: Theory and Applications (USYS), Penang,
Malaysia, 13–14 December 2016; pp. 19–25. [CrossRef] 64. Yu, P.; Zhao, Y.; Wang, T.; Zhou, H.; Su, S.; Zhen, C. Steady-state spiral motion simulation and turning speed
analysis of an underwater glider. In Proceedings of the 2017 4th International Conference on Information
Cybernetics and Computational Social Systems (ICCSS), Dalian, China, 24–26 July 2017; pp. 372–377. 65. Bloch, T.; Krishnaprasad, P.S.; Leonard, N.; Murray, R. Jerrold Eldon Marsden [Obituaries]. Control Syst. IEEE 2011, 31, 105–108. 66. Williams, C.D. AUV systems research at the NRC-IOT: An update. In Proceedings of the Underwater
Technology 2004—UT ’04. 2004 International Symposium on, Taipei, Taiwan, 20–23 April 2004; pp. 59–73. 67. Jing, D.; Haifeng, W. References [CrossRef] 48. Leonard, N.E.; Graver, J.G. Model-Based feedback control of autonomous underwater gliders. IEEE J. Ocean
Eng. 2001, 26, 4. [CrossRef] 48. Leonard, N.E.; Graver, J.G. Model-Based feedback control of autonomous underwater gliders. IEEE J. Ocean
Eng. 2001, 26, 4. [CrossRef] 49. Arima, M.; Ichihashi, N.; Ikebuchi, T. Motion characteristics of an underwater glider with independently
controllable main wings. In Proceedings of the OCEANS 2008—MTS/IEEE Kobe Techno-Ocean, Kobe, Japan,
8–11 April 2008; pp. 1–7. 49. Arima, M.; Ichihashi, N.; Ikebuchi, T. Motion characteristics of an underwater glider with independently
controllable main wings. In Proceedings of the OCEANS 2008—MTS/IEEE Kobe Techno-Ocean, Kobe, Japan,
8–11 April 2008; pp. 1–7. Sensors 2019, 19, 1831 16 of 17 16 of 17 50. Roger, E.O.; Genderson, J.G.; Smith, W.S.; Denny, G.; Farley, P.J. Underwater acoustic glider. In Proceedings of
the IEEE International Geoscience and Remote Sensing Symposium, Anchorage, AK, USA, 20–24 September
2004; pp. 2241–2244. 51. Woithe, H.C.; Kremer, U. A programming architecture for smart autonomous underwater vehicles. In Proceedings of the IEEE/RSJ International Conference on Intelligent Robots and Systems 2009—IROS 2009,
St. Louis, MI, USA, 11–15 October 2009; pp. 4433–4438. 52. Woithe, H.C.; Tilkidjieva, D.; Kremer, U. Towards a Resource-Aware Programming Architecture for Smart
Autonomous Underwater Vehicles; Technical Report, DCS-TR-637; Rutgers University: New Brunswick, NJ,
USA, 2008. 53. Lennart, L. System Identification Theory for the User, 2nd ed.; Prentice Hall: Upper Saddle River, NJ, USA, 1999. 53. Lennart, L. System Identification Theory for the User, 2nd ed.; Prentice Hall: Upper Saddle River, NJ, USA, 1999. 54
Petritoli E ; Leccese F High accuracy attitude and navigation system for an autonomous underwater vehicle 53. Lennart, L. System Identification Theory for the User, 2nd ed.; Prentice Hall: Upper Saddle River, NJ, USA, 1999. 54. Petritoli, E.; Leccese, F. High accuracy attitude and navigation system for an autonomous underwater vehicle
(AUV) A t IMEKO 2018 7 3 9 [C
R f] 54. Petritoli, E.; Leccese, F. High accuracy attitude and navigation system for an autonomous underwater vehicle
(AUV). Acta IMEKO 2018, 7, 3–9. [CrossRef] 55. Petritoli, E.; Leccese, F. A high accuracy navigation system for a tailless underwater glider. In Proceedings
of the IMEKO TC19 Workshop on Metrology for the Sea, MetroSea 2017: Learning to Measure Sea Health
Parameters, Naples, Italy, 11–13 October 2017; pp. 127–132. 56. Petritoli, E.; Leccese, F. A high accuracy attitude system for a tailless underwater glider. 69.
Vogeltanz, T. A Survey of Free Software for the Design, Analysis, Modelling, and Simulation of an Unmanned
Aerial Vehicle. Arch. Comput. Meth. Eng. 2016, 23, 449. [CrossRef] References System health management for unmanned aerial vehicle: conception, state-of-art,
framework and challenge. In Proceedings of the Electronic Measurement & Instruments (ICEMI). In Proceedings of the 2013 IEEE 11th International Conference, Harbin, China, 16–19 August 2013; Volume 2,
pp. 859–863. 68. Javafoil—Analysys of Airfolis. Available online: https://www.mh-aerotools.de/airfoils/javafoil.htm (accessed
on 15 February 2019). 69. Vogeltanz, T. A Survey of Free Software for the Design, Analysis, Modelling, and Simulation of an Unmanned
Aerial Vehicle. Arch. Comput. Meth. Eng. 2016, 23, 449. [CrossRef] 17 of 17 Sensors 2019, 19, 1831 17 of 17 70. Vogeltanz, T.; Jašek, R. Free software for the modelling and simulation of a mini-UAV. In Proceedings of the
Mathematics and Computers in Science and Industry, Varna, Bulgaria, 13–15 September 2014; pp. 210–215. 71. Jodeh, N.; Blue, P.; Waldron, A. Development of small unmanned aerial vehicle research platform: Modeling
and simulating with flight test validation. In Proceedings of the AIAA Modeling and Simulation Technologies
Conference and Exhibit, Keystone, CO, USA, 21–24 August 2006; p. 6261. 72. Mueller, T.J.; DeLaurier, J.D. Aerodynamics of small vehicles. Annual review of fluid mechanics 2003, 35, 89–111. [CrossRef] 73. Fabiani, P.; Fuertes, V.; Piquereau, A.; Mampey, R.; Teichteil-Königsbuch, F. Autonomous flight and navigation
of VTOL UAVs: From autonomy demonstrations to out-of-sight flights. Aerosp. Sci. Technol. 2007, 11,
183–193. [CrossRef] 74. Wu, N.; Wu, C.; Ge, T.; Yang, D.; Yang, R. Pitch Channel Control of a REMUS AUV with Input Saturation and
Coupling Disturbances. Appl. Sci. 2018, 8, 253. [CrossRef] 75. Hahn, A. Vehicle sketch pad: A parametric geometry modeller for conceptual aircraft design. In Proceedings
of the 48th AIAA Aerospace Sciences Meeting Including the New Horizons Forum and Aerospace Exposition,
Orlando, FL, USA, 4–7 January 2010; p. 657. [CrossRef] 76. Wang, T.; Wu, C.; Wang, J.; Ge, T. Modeling and Control of Negative-Buoyancy Tri-Tilt-Rotor Autonomous
Underwater Vehicles Based on Immersion and Invariance Methodology. Appl. Sci. 2018, 8, 1150. [CrossRef] 77. Boussalis, H.; Valavanis, K.; Guillaume, D.; Pena, F.; Diaz, E.U.; & Alvarenga, J. Control of a simulated
wing structure with multiple segmented control surfaces. In Proceedings of the 2013 21st Mediterranean
Conference IEEE Control & Automation (MED), Crete, Greece, 25–28 June 2013; pp. 501–506. 78. Wang, T.; Wang, J.; Wu, C.; Zhao, M.; Ge, T. Disturbance-Rejection Control for the Hover and Transition
Modes of a Negative-Buoyancy Quad Tilt-Rotor Autonomous Underwater Vehicle. Appl. Sci. 2018, 8, 2459. [CrossRef] © 2019 by the authors. Licensee MDPI, Basel, Switzerland. References This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/). © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).
|
W2324349643.txt
|
http://www.nomos-elibrary.de/10.5771/0340-8574-2007-1-29.pdf
|
de
|
Das Format mit Inhalt füllen
|
Blätter der Wohlfahrtspflege
| 2,007
|
cc-by
| 782
|
Monitoring
Bei Abschnitt C (Planung und Konzeption) geht es nicht nur um die Vorbereitung von Interventionen (»Dienstleistungen«), sondern auch von Hilfesystemen
und um die Prozessplanung. Auch hier
unterscheiden sich Bachelor und Master
durch den Grad der Eigenständigkeit und
hinsichtlich der Fähigkeit, Lösungsstrategien für neue und unbekannte Aufgabenstellungen zu entwickeln.
Der Abschnitt D (»Recherche und Forschung in der Sozialen Arbeit«) hat die
deutlichste Wissenschaftsorientierung,
denn es geht für Bachelor und Master um
die Bearbeitung von Forschungsfragen
und um fachliche Informationsbeschaffung. Master werden breitere Kenntnisse
und weit mehr Eigenständigkeit als dem
Bachelor attestiert Während Bachelor
beispielsweise die Fähigkeit zugeordnet
ist, angeleitete Praxisforschung zu betreiben, verfügen Master über die Fähigkeit,
das Design von Untersuchungen zu entwickeln, Forschungsergebnisse kritisch zu
bewerten und innovative Methoden und
Strategien zu entwickeln.
Das Format mit Inhalt füllen
Die Deutsche Gesellschaft für Soziale Arbeit versteht sich als unabhängiges Forum
für die Wissenschaft und Praxis Sozialer Arbeit. Sie nimmt für sich in Anspruch, die
Sache der Profession der Sozialen Arbeit und die Disziplin der Sozialarbeitswissenschaft zu vertreten. Ihr Vorsitzender Prof. Dr. Wolf Rainer Wendt äußert sich in einem Kurzinterview zu dem im letzten Jahr verabschiedeten Qualifikationsrahmen
Soziale Arbeit.
Der Abschnitt E (»Organisation,
Durchführung und Evaluation in der Sozialen Arbeit«) behandelt die sichtbarste
Ebene der Sozialen Arbeit: von ihrer Organisation und Durchführung bis hin zu
ihrer Evaluation: Einschätzung und Einsatz
sächlicher
und
personeller
Ressourcen. Für Master wird u. a. festgestellt, dass sie auch die Weiterentwikklung von Methoden und Qualitätsmanagementsystemen betreiben können.
Der Abschnitt F schließlich (»Professionelle allgemeine Fähigkeiten und Haltungen in der Sozialen Arbeit«) benennt »weitere, nicht fachspezifische Fähigkeiten, die
für die erfolgreiche, professionelle Soziale
Arbeit als Vorbedingung gelten müssen«.
Der Abschnitt G (»Persönlichkeit und Haltungen«) differenziert bewusst nicht zwischen Bachelor und Master.
Fazit und Ausblick
Blätter der Wohlfahrtspflege: Wie steht die Deutsche Gesellschaft für Soziale Arbeit
zu dem Qualifikationsrahmen des Fachbereichstages?
Wolf Rainer Wendt: Unsere Gesellschaft begrüßt diese Ausarbeitung. Wichtig ist die
grundsätzliche Entscheidung für ein generalistisches Studium. Die Darstellung
kommt einem wissenschaftlichen Anspruch nach, den die Soziale Arbeit wie andere
Disziplinen in der Lehre an Hochschulen erheben kann und formuliert diesen Anspruch auf den Qualifikationsniveaus des Bachelor- und des Masterstudiums. Das
Format, das den Studiengängen zugeschrieben wird, bedarf aber einer inhaltlichen
Ausfüllung. Im Text wird von einer »Prozesslogik« Sozialer Arbeit sowie von einem
»kollektiven Wissens- und Fähigkeitskanon« gesprochen – ohne nähere Angaben
zur Sache.
Blätter der Wohlfahrtspflege: Welcher Inhalt passt Ihrer Ansicht nach in den Qualifikationsrahmen?
Wolf Rainer Wendt: Wir haben zur inhaltlichen Ausgestaltung des Studiums der Sozialen Arbeit ein Kerncurriculum zur Diskussion gestellt (nachzulesen auf unserer
Website http://www.dgsinfo.de, Rubrik Mitteilungen, Online-Mitteilung 77). Die
Hochschulausbildung muss den ganzen Gegenstandsbereich der Sozialen Arbeit auf
dem Bachelor-Level behandeln. Die Berufsbefähigung, die ein Bachelor haben sollte, hängt weniger von der Fülle des Wissens aus herkömmlichen »Bezugsdisziplinen«
als von der Kompetenz ab, die in der Behandlung der Problematik gesellschaftlicher
Daseinsgestaltung und individueller Lebensführung in konkreten personen- und situationsbezogenen Aufgabenstellungen erworben wird. Einen breiten Einblick in die
Praxis eingeschlossen. Das Masterstudium baut auf dieser Fundierung, aber nicht
durch »Mehr vom Gleichen«, sondern mit der Kompetenz für Entwicklungs- und
Steuerungsaufgaben.
Blätter der Wohlfahrtspflege: Wie müssen nun die nächsten Schritte aussehen?
Wolf Rainer Wendt: Zu klären ist die Differenzierung der Masterstudiengänge. Die
Profession braucht einerseits einen sozialarbeitswissenschaftlich akzentuierten, stärker forschungsorientierten konsekutiven Masterstudiengang, der für Leitungsaufgaben qualifiziert und der (in Verbindung mit oder an einer Universität) die Voraussetzungen für eine Promotion schafft und so in Zukunft eine Selbstrekrutierung der
Lehre im Fachgebiet ermöglicht. Andererseits ist darauf hinzuwirken, dass für die
Spezialisierungen im Handlungsfeld solche anwendungsbezogenen Studiengänge
vorhanden sind, die für eine Fachsozialarbeit qualifizieren, damit der Profession zugeordnet bleiben und nicht den Zusammenhang mit ihr verlieren.
Blätter der Wohlfahrtspflege 1/2007
Der Qualifikationsrahmen Soziale Arbeit ist – nach Modularisierung und Umstellung auf Bachelor und Master – eine
weitere Mitwirkung der Fachbereiche der
Sozialen Arbeit an der Harmonisierung
des europäischen Hochschulraumes und
der Abstimmung von hochschulisch und
außerhochschulisch erworbenen beruflichen Qualifikationen.
Der Qualifikationsrahmen Soziale Arbeit ist kein berufspolitisches Dokument.
Man könnte ihn als Ausbildungsversprechen bezeichnen, das den Mindestumfang
der mit dem Abschluss Bachelor oder
Master erworbenen und geprüften Qualifikationen umfasst. Er trifft keine inhaltliche Festlegung und stellt keine endgültige und vollständige Liste einzelner professioneller Kompetenzen vor. Im
Wettstreit der Disziplinen kann er der Sozialen Arbeit als legitimatorisches Instrument dienen – beispielsweise um gute Zugangsmöglichkeiten zur Promotion.
Die Anregungen und Kommentare, die
bis hin zur Abschlussdiskussion vorgelegt
und behandelt wurden, haben wesentlich
zur Qualität des Qualifikationsrahmens
Soziale Arbeit beigetragen. In der einstimmigen (!) Beschlussfassung wurde
eine Revision des Qualifikationsrahmens
Soziale Arbeit in eineinhalb Jahren vorgesehen. Eine fraglos wichtige Weiterentwicklung wäre die Hinzunahme der Doktoratsebene. Diskutiert wird auch, wie die
Anschlussstellen an internationale Doku-
https://doi.org/10.5771/0340-8574-2007-1-29
Generiert durch IP '51.159.168.146', am 29.06.2024, 12:26:14.
Das Erstellen und Weitergeben von Kopien dieses PDFs ist nicht zulässig.
29
|
|
https://openalex.org/W2995501892
|
https://www.scielo.br/j/rgo/a/VV9Kn9QVddchL4tHB8xcCTd/?lang=en&format=pdf
|
English
| null |
Efficacy assessment of oral and sublingual sedation using alprazolam in implantology surgical procedures
|
RGO - Revista Gaúcha de Odontologia
| 2,019
|
cc-by
| 5,627
|
Efficacy assessment of oral and sublingual sedation using
alprazolam in implantology surgical procedures Avaliação da eficácia da sedação por via oral e sublingual com
alprazolam em procedimentos cirúrgicos de Implantodontia Antonio José Bittencourt da ROSA1
Bruno Salles SOTTO-MAIOR2 0000-0002-9462-0299
Carlos Eduardo FRANCISCHONE3
Juliana Cama RAMACCIATO4 0000-0002-3081-1504
Rui Barbosa de BRITO JÚNIOR5 0000-0002-3484-9438
Rogério Heládio Lopes MOTTA4 0000-0002-6983-7883 Antonio José Bittencourt da ROSA1
Bruno Salles SOTTO-MAIOR2 0000-0002-9462-0299
Carlos Eduardo FRANCISCHONE3
Juliana Cama RAMACCIATO4 0000-0002-3081-1504
Rui Barbosa de BRITO JÚNIOR5 0000-0002-3484-9438
Rogério Heládio Lopes MOTTA4 0000-0002-6983-7883 NICAL | CLÍNIC
| ORIGINAL NICAL | CLÍNIC
| ORIGINAL http://dx.doi.org/10.1590/1981-86372019000603516 Efficacy assessment of oral and sublingual sedation using
alprazolam in implantology surgical procedures Rosa AJB, Sotto-Maior BS, Francischone CE, Ramacciato JC, Brito Junior RB, Motta RHL Efficacy assessment of oral and sublingual sedation
using alprazolam in implantology surgical procedures. RGO, Rev Gaúch Odontol. 2019;67:e20190060. http://dx.doi.org/10.1590/1981-
86372019000603516 g
p
g
SP, Brasil.
4 Faculdade São Leopoldo Mandic, Instituto de Pesquisas São Leopoldo Mandic, Área de Farmacologia, Anestesiologia e Terapêutica.
Rua José Rocha Junqueira, 13, 13045-755, Campinas, SP, Brasil. Correspondência para / Correspondence to: RHL MOTTA. E-mail:
<rogerio.motta@slmandic.edu.br>. RESUMO Objetivo: Avaliar a eficácia de duas formulações de alprazolam (ALP) na sedação de voluntários submetidos à procedimentos cirúrgicos
de Implantodontia. Métodos: Foi realizado um estudo clinico, randomizado e cego, com 52 voluntários (ambos os gêneros) que
foram divididos em 2 grupos: G1 (n=26) - administração de ALP (0,5mg) por via oral uma hora antes da sessão clínica; e G2 (n=26) -
administração de ALP (0,5mg) por via sublingual 30 minutos antes da sessão clínica. Foram avaliadas a pressão arterial (PA) e frequência
cardíaca (FC), e foram utilizadas a escala de ansiedade dental de Corah (COR) e escala analógica visual (EAV) para avaliação do nível de
ansiedade. Adicionalmente, foram utilizados o teste de Trieger (TRI) e um questionário de percepção (QP) para avaliar as respostas aos
tratamentos. Todos os parâmetros foram avaliados em 3 momentos (pré, trans e pós-operatório imediato). Resultados: Para PA e FC
não foram observadas diferenças estatísticas significativas entre os períodos para G1 e G2 (p>0,05). Para COR, a maioria dos voluntários
foi classificada como ansiosos ou muitos ansiosos em ambos os grupos (p>0,05), e para EAV a diminuição da ansiedade foi significativa
(p<0,05) e não houve diferença estatistica significativa entre os dois grupos tanto no período “pré” (p=0,18) quanto no “pós” (p=0,12). Para TRI, não houve diferença significativa entre os períodos pré e pós considerando o número de pontos perdidos (p>0,05). Quanto
ao QP, ambas as formulações tiveram boa aceitabilidade pelos voluntários (p>0,05), não ocorrendo amnésia anterógrada. Conclusão:
Concluiu-se que ambas as formulações de alprazolam foram eficazes para a sedação dos voluntários avaliados. studied and recommended for dental procedures in
anxious patients [10,11]. Taking into consideration the
age range of patients treated in the area of Implantology,
alprazolam may be a choice of treament due to its latency
time and shorter half-life [11]. In addition, one research
compared alprazolam pharmacokinetic and farmacodynamic
parameters in the elderly and young adults, with only a
small increase in plasma concentration in the elderly with
no evidence of an increase in the effect of the drug [12]
Meanwhile, oral administration of alprazolam can present
some limitations, such as variable action time and effect
delay [13]. Literature has discussed the search for new
anxiolytics suggesting another sublingual formulation
of alprazolam, which presents a shorter latency time as
clinical advantage. ABSTRACT Objective: The aim of this study was to evaluate the efficacy of two formulations of alprazolam (ALP) in patients undergoing oral
implantology surgical procedures. Methods: A single-blinded randomized clinical study was performed with 52 volunteers (both
genders) divided into 2 groups: G1 (n = 26) – oral administration of ALP (0.5 mg) 1h before surgical procedures; and G2 (n = 26) –
sublingual administration of ALP (0.5mg) 30 minutes before surgery. Blood pressure (BP) and heart rate (HR) were evaluated. Corah
dental anxiety scale (COR), visual analogic scale (VAS), Trieger test (TRI) and the perception questionnaire (PQ) were used to assess
the level of anxiety and responses to formulations, respectively. All parameters were evaluated at three times (be-fore, during and
postoperative). Results: For BP and HR no statistically significant differences were observed between periods for G1 and G2 (p> 0.05). For COR, most volunteers were classi-fied such as anxious or very anxious. For VAS, decreased anxiety was significant (p< 0.05) and no
statistically significant differences between the two groups both in the period “pre” (p= 0.18) and “post” (p= 0.12) were observed. For
TRI, there were no significant differences between the periods considering the number of points lost (p> 0.05). The anxiety reduction
was significant (p< 0.05) for both formulations. As for PQ, both formulations showed good volunteers’ acceptance (p> 0.05), and
they have not reported anterograde amnesia. Conclusion: It was concluded that both formulations were effective for sedation of the
evaluated volunteers. Indexing terms: Alprazolam. Conscious sedation. Dental implantation. poldo Mandic, Instituto de Pesquisas São Leopoldo Mandic, Área de Histologia. Campinas, SP, Brasil. Rosa AJB, Sotto-Maior BS, Francischone CE, Ramacciato JC, Brito Junior RB, Motta RHL Efficacy assessment of oral and sublingual sedation
using alprazolam in implantology surgical procedures. RGO, Rev Gaúch Odontol. 2019;67:e20190060. http://dx.doi.org/10.1590/1981-
86372019000603516 CC
BY RGO, Rev Gaúch Odontol. 2019;67:e20190060 AJB ROSA et al. RGO, Rev Gaúch Odontol. 2019;67:e20190060 Objetivo: Avaliar a eficácia de duas formulações de alprazolam (ALP) na sedação de voluntários submetidos à procedimentos cirúrgicos
de Implantodontia. Métodos: Foi realizado um estudo clinico, randomizado e cego, com 52 voluntários (ambos os gêneros) que
foram divididos em 2 grupos: G1 (n=26) - administração de ALP (0,5mg) por via oral uma hora antes da sessão clínica; e G2 (n=26) -
administração de ALP (0,5mg) por via sublingual 30 minutos antes da sessão clínica. Foram avaliadas a pressão arterial (PA) e frequência
cardíaca (FC), e foram utilizadas a escala de ansiedade dental de Corah (COR) e escala analógica visual (EAV) para avaliação do nível de
ansiedade. Adicionalmente, foram utilizados o teste de Trieger (TRI) e um questionário de percepção (QP) para avaliar as respostas aos
tratamentos. Todos os parâmetros foram avaliados em 3 momentos (pré, trans e pós-operatório imediato). Resultados: Para PA e FC
não foram observadas diferenças estatísticas significativas entre os períodos para G1 e G2 (p>0,05). Para COR, a maioria dos voluntários
foi classificada como ansiosos ou muitos ansiosos em ambos os grupos (p>0,05), e para EAV a diminuição da ansiedade foi significativa
(p<0,05) e não houve diferença estatistica significativa entre os dois grupos tanto no período “pré” (p=0,18) quanto no “pós” (p=0,12).
Para TRI, não houve diferença significativa entre os períodos pré e pós considerando o número de pontos perdidos (p>0,05). Quanto
ao QP, ambas as formulações tiveram boa aceitabilidade pelos voluntários (p>0,05), não ocorrendo amnésia anterógrada. Conclusão:
Concluiu-se que ambas as formulações de alprazolam foram eficazes para a sedação dos voluntários avaliados.
d
d
l
l
d
l
d Termos de indexação: Alprazolam. Sedação consciente. Implantação dentária. METHODS anxiety”. The classification was performed by placing a
vertical mark on the line and the distance between the
mark and the 0 end was measured by a digital caliper
(Mitutoyo®, USA), thus assessing the volunteer’s anxiety
level [18]. This research was submitted and approved by the
Research Ethics Committee of Faculdade São Leopoldo
Mandic under protocol CAAE# 30650314.3.0000.5374. The sample of this research consisted of 52 (fifty-two)
volunteers, of both genders, mean age of 51.8 (± 7.1)
years, who sought for oral rehabilitation care. After the surgical procedure, the volunteers were
also asked to respond to a questionnaire in order to
evaluate their perception regarding the sedation method,
the occurrence of anterograde amnesia and the presence
of side effects. In addition, the Trieger test was used, which
consists of joining points of a pre-established figure in two
moments: in the immediate preoperative period and after
the end of the procedures [19,20]. Additional information
on surgical procedures was also recorded, such as number
of implants installed, the place of the installation, time
of surgery, number of anesthetic cartridge used and any
complications during surgery. The study was performed in patients requiring
surgical intervention to install at least four implants in the
same surgery or maxillary sinus surgeries. In the routine
treatment of anxiety, alprazolam is used in doses of 0.5
to 0.75mg in adults and 0.25 to 0.5mg in the elderly [15]. Considering the doses available in the Brazilian market for
both pharmaceutical formulations (oral and sublingual),
the dose of 0.5mg was adopted for both formulations in
the present study. Based on the above, the volunteers were
divided into 2 groups: group 1 (n = 26) – oral administration of
alprazolam 0.5 mg tablet (Frontal®, Pfizer) one hour before
the clinical procedure, and group 2 (n = 26) – sublingual
administration of alprazolam 0.5mg tablet (Frontal SL®,
Pfizer) 30 minutes before the clinical procedure. For surgical procedures, local anesthesia was
performed with lidocaine 2% with epinephrine 1: 100,000
(Alphacaine®, Nova DFL) for blocking the involved nerves
or infiltrative technique in each case. The number of
anesthetic cartidrges were recorded in the clinical form
of each volunteer. Prophylactic antibiotic therapy (oral
administration of 2g of amoxicillin (Medley®) and oral
single dose of 4 mg dexamethasone (Decadron®, Aché)
was administered one hour before the procedures as
a drug protocol for all implant procedures. INTRODUCTION Fear and anxiety are routine problems in the dental
clinic and dental treatment can be considered a crucial
problem for odontophobic patients [1]. Fear of undergoing
dental treatments or anxiety about these treatments is
called by several authors as dental anxiety or odontophobia
[2]. Literature has presented several terms concerning the
patient’s acceptance of the dental treatment, such as the
fear of the treatment itself and dental anxiety, which is
related to the fear of seeking and attending a dental office
and phobia [3]. Fear and anxiety in dental treatment is worldwide. However, some subgroups present higher levels than others
[4] which may lead to dental appointment avoidance,
or yet only seek care in cases of pain [5]. Furthermore,
odontophobic patients may be less cooperative during
procedures and present less satisfaction with surgical
treatments [6] Within this context, a study has shown that
sublingual alprazolam dosage obtained a higher plasma
concentration peak (17.3 vs 14.9 ng/ml) and a shorter
time to reach it (1.17 vs 1.73 hours) when compared to
the same orally taken dose (1mg) [14]. It was concluded
that alprazolam sublingual absorption is fast and a
viable alternative for people with panic syndrome. For
dental procedures, these drug features would allow
its administration in a shorter time interval during the
preoperative as well as in a controlled environment. However, further studies should be developed in order
to evaluate the drug efficacy in Dentistry, mainly in
Implantology. Thus, the aim of this study was to compare
the efficacy of oral and sublingual alprazolam sedation in
volunteers undergoing implantology surgical procedures. Dental surgical procedures such as the installation
of dental implants are associated with high pain
expectation, one of the main causes of anxiety [7] which
can impair the pain control and trigger the need for larger
amounts of anesthetics, analgesics and other medications
[8]. Thus, it is of utmost importance that anxiety caused
by dental treatment is reduced as well as other factors
associated with anxiety such as pain expectations and
previous negative experiences related to pre-surgical and
trans-surgical anxiety [9]. Among the clinical options for anxiety control in
Dentistry, oral sedation with benzodiazepines has been 2 Oral and sublingual sedation METHODS RGO, Rev Gaúch Odontol. 2019;67:e20190060 METHODS To control
postoperative pain, ibuprofen 400mg (Alivium®, Mantecorp)
was administered every six hours for 2 days. For the other
surgical procedures performed, the antibiotic prophylaxis
was performed according to the patient’s profile and
procedure invasiveness [21,22]. The volunteers received detailed information about
the study in order to obtain the informed consent form
and were included according to the following inclusion
and exclusion criteria: a) treatment plan accepted and in
need of surgical procedures; b) volunteers classified as
healthy during clinical anamnesis (ASA I or ASA II); c) ability
to provide written consent; d) not use benzodiazepines or
analgesics for a period of at least 15 days before surgeries;
e) no contraindication to the use of the drugs proposed
in the study (allergy, glaucoma, myasthenia gravis,
pregnancy, among others). After volunteer’s agreement of
participation and signing of the consent term, a careful
anamnesis was performed followed by a clinical chart
completion and the volunteer’s anxiety was evaluated by
the Corah scale [16,17]. The results were initially submitted to Shapiro-Wilk
and Bartlett test in order to observe the normality of the
distribution and the similarity of the deviations, respectively. Normal distribution or similar deviations (p <0.05) were not
observed, and the data were submitted to non-parametric
tests. The relative distribution of age, gender and other
factors according to the studied groups was evaluated by
the Mann-Whitney and Chi-square test. Afterwards, the volunteers’ vital signs (heart and
respiratory rate, blood pressure and oxygen saturation)
were monitored during the procedures at different
moments (immediate preoperative period, after the first
dental implant was installed or after access to the maxillary
sinus, end of the procedure). Anxiety was also measured
by visual analogue scale (VAS) before and after procedures. The VAS is characterized by a 10 cm straight line with no
numbers except at the ends where 0 (zero) corresponds
to “no anxiety” and 10 (ten) to “maximum intensity of Systolic and diastolic pressures, heart rate over
time and the influence of treatments for each group
(VAS values) were separately assessed by Friedman test,
and the comparison between the groups was performed
by the Mann-Whitney test. Trieger test parameters were
evaluated by the Wilcoxon test for comparison between RGO, Rev Gaúch Odontol. 2019;67:e20190060 3 AJB ROSA et al. the pre and final periods and by Mann-Whitney test for
evaluation between the groups. METHODS The volunteers’ perception
of the effect of the treatments was analyzed using Fisher’s
exact test and Chi-square test. All analyzes were performed
considering a significance level of 5%, using GraphPad
Prism 6.0 and BioEstat 5.0 softwares. the procedure performed in each group did not present
significant differences between the groups leading to
sample homogeinity. The volunteers’ distribution in each
group considering the classification of anxiety, measured by
Corah scale and age, was also homogeneous. Figure 1 shows the results obtained for systolic
and diastolic blood pressure in relation to the administered
formulations. RESULTS It was observed that the systolic pressure decreased
(Friedman test, p< 0.05) over time for both groups. In
fact, the mean difference (± standard deviation) between
the measurements obtained in the period pre and pos
showed a 5% reduction in the systolic pressure. The mean
difference between the pre and final periods was -6.8% As shown in table 1, no statistically significant
differences (p> 0.05) between the groups concerning
the surgical time required for the procedures, amount of
local anesthetic used, gender and the systemic profile of
the volunteers. Moreover, the type and characteristics of Table 1. Relative distribution of gender and other factors according to the studied groups. Factors
Group 1 (oral) n=26
Group 2 (sublingual) n=26
P values
Surgical time in hours
(mean ± standard deviation)
2.2 (±0.8)
2.0 (±0.5)
0.52
Number of cartridges used
(mean ± standard deviation)
3.5 (±0,6)
3.7 (±0,6)
0.43
Gender
Male
10 (38.4%)
10 (38.4%)
1.00
Female
16 (61.6%)
16 (61.6%)
ASA
I
20 (76.9%)
21 (80.8%)
0.73
II
6 (23.1%)
5 (19.2%)
Procedure
Only sinus lift
9 (34.6%)
9 (34.6%)
1.00
Only implants
15 (57.7%)
15 (57.7%)
Implants + sinus lift
2 (7.7%)
2 (7.7%)
Operated side
Both
7 (26.9%)
7 (26.9%)
0.26
Right
8 (30.8%)
13 (50%)
Left
11 (42.3%)
6 (23.1%)
Region
Mandíbule
9 (34.6%)
10 (38.5%)
0.77
Maxillae
17 (65.4%)
16 (61.5%)
Anterior
5 (19.2%)
5 (19.2%)
1.00
Posterior
19 (73.1%)
19 (73.1%)
Both
2 (7.7%)
2 (7.7%)
Corah Scale
Not anxious
8 (30.8%)
7 (26.9%)
0.14
Anxious
7 (26.9%)
2 (7.7%)
Very anxious
7 (26.9%)
7 (26.9%)
Phobic
4 (15.4%)
10 (38.5%) Table 1. Relative distribution of gender and other factors according to the studied groups. Oral and sublingual sedation Figure 1. Median (interquartile range) of systolic arterial pressures (filled lines) and diastolic (dashed lines) according to groups and verification periods. rtile range) of systolic arterial pressures (filled lines) and diastolic (dashed lines) according to groups and verification periods. Figure 1. Median (interquartile range) of systolic arterial pressures (filled lines) and diastolic (dashed lines) according to grou (± 7.1%) and -9.1% (± 7.8%), respectively for groups
1 and 2. In addition, the comparison among the mean
differences showed no statistically significant differences
(Kruskal-Wallis test, p>0.05) between the groups. RESULTS It was
also observed that, considering the systolic pressure found
before (p = 0.61), trans (p = 0.65) or at the end of the
procedures (p = 0.87), there was no statistically significant
difference Mann-Whitney between the two groups. Regarding diastolic blood pressure, it was
observed that for both groups there was no significant
statistical difference between the periods (Friedman test,
p> 0.05). Furthermore, no significant statistical differences
(Mann-Whitney test) were observed between the two
groups, considering the diastolic pressure before (p= 0.53),
trans (p= 0.90) or at the end of the procedures (p= 0.78). Actually, the differences between the diastolic pressures Figure 2. Median (interquartile range) of heart rate according to groups and periods of verification. Figure 2. Median (interquartile range) of heart rate according to groups and periods of verification. Figure 2. Median (interquartile range) of heart rate according to groups and periods of verification RGO, Rev Gaúch Odontol. 2019;67:e20190060 AJB ROSA et al. Trieger test, as well as by the number of lost points (figure
5) when filling the same test. There were no statistical
significant differences (Wilcoxon test) between the pre
and final periods, considering the number of points lost
for both group 1 (p= 0.07) and group 2 (p= 0.06) and
neither between groups 1 and 2 (Mann-Whitney test),
both for the pre (p= 0.48) and final (p= 0.92) periods. However, the time spent filling the test was significantly
lower (Wilcoxon’s test) in the final period for both group 1
(p= 0.01) and group 2 (p< 0.0001), although no statistical
significant differences (Mann-Whitney test) were found
between the groups, both in the pre- (p= 0.84) and in the
post-procedure periods (p= 0.29). measured in the pre-period and the others showed a 1.5%
reduction or less. Thus, there was no significant blood
pressure change for both alprazolam formulations. Figure 2 shows the effects of the two treatments
over the periods evaluated regarding heart rate. It was observed
that there was a decrease (Friedman test, p< 0.05) over time,
for both groups. Nonetheless there was no significant
statistical difference (Mann-Whitney test) between the
two groups either before (p= 0.95), during (p= 0.41)
or at the end of the procedures (p= 0.93). Thus, there
was no difference between the effects of treatments on
heart rate. Figure 3 shows the influence of treatments on
anxiety measured by VAS in two different periods. RESULTS The anxiety
reduction was very significant (Friedman test, p< 0.0001)
between the “pre” and “final” periods for groups 1 and 2. However, there was no statistical significant difference
between the two groups in both “pre” (p= 0.18)
and “final” (p= 0.12) periods. Thus, both treatments
caused significant reduction of anxiety (measured by
VAS). Nonetheless, no difference was found between
alprazolam formulations. Table 2 shows the volunteers’s perception
regarding the different forms of alprazolam effects. The
analysis of the data revealed that there was no statistically
significant difference between the treatments in relation to
the procedure recall and most of the volunteers reported
that remembered the whole procedure. Moreover,
although no differences were observed between the
groups regarding previous experiences, most volunteers
pointed out that they felt a lot of difference compared to
previous dental procedures. More drowsiness was reported
in the volunteers of group 2 than in group 1. The treatment effects on individuals’ alertness
was assessed by the time (figure 4) spent to complete the Figure 3. Median (interquartile range) of anxiety (measured by VAS) according to the groups and the periods assessed. Fi
3 M di
(i
il
)
f
i
(
d b VAS)
di
h
d h
i d
d Figure 3. Median (interquartile range) of anxiety (measured by VAS) according to the groups and the periods assessed. Oral and sublingual sedation Oral and sublingual sedation Figure 4. Number of lost points during the completion of the Trieger test according to the groups and the evaluation periods. The center line represents the
median, the box represents the 1st and 3rd quartiles and the whiskers show the minimum and maximum values. Figure 4. Number of lost points during the completion of the Trieger test according to the groups and the evaluation periods. The center line represents the
median, the box represents the 1st and 3rd quartiles and the whiskers show the minimum and maximum values. Figure 4. Number of lost points during the completion of the Trieger test according to the groups and the evaluation periods. The center line represents the
median, the box represents the 1st and 3rd quartiles and the whiskers show the minimum and maximum values. Figure 5. Time spent (in seconds) during the completion of the Trieger test according to the groups and the verification periods. DISCUSSION Benzodiazepines are commonly drugs used orally
in Dentistry due to their efficacy and safety, since it has an
effective reverser and present good patient acceptability
[10,13,21]. Nonetheless, there have been few clinical
studies concerning the sublingual use of these drugs in
dental procedures, which has led the present study. In the present study, the anxiety reduction between
the pre- and postoperative period was also very significant
with the administration of both alprazolam formulations,
suggesting that the two formulations were effective in
the anxiety control of the evaluated volunteers. This result
corroborates with Wolf et al. [30] who observed anxiety
reduction in volunteers with doses of 0.25mg and 1mg
of alprazolam when compared to placebo. In addition,
systolic and heart rate values were decreased and diastolic
pressure remained stable, according to Joshi et al. [31],
who observed the stability of vital signs and reduction of
anxiety for the dose of 0.5mg for pre-medication in oral
surgery. Regarding drug administration, sublingual alprazolam
is expected to provide a faster action onset and greater
bioavailability than oral administration due to its rapid
absorption and first passmetabolism avoidance. Scavone
et al. [20] compared alprazolam when administered orally
or sublingually and found a higher plasma concentration
with the sublingual route, although the differences have
not shown statistical significance. The benzodiazepine
sublingual administration has proved better intake over
oral administration due to its increased bioavailability and
to faster promote higher blood concentration [23-25]. A possible hypothesis for this arterial pressure
decrease would be the more controlled response of
adrenaline and noradrenaline during the stress, confirming
the findings of Van den Berg et al. [32], who suggested that
alprazolam presents specific adrenomedullary suppression
activity. These results demonstrate the clinical safety of
alprazolam at the dose of 0.5 mg in the preoperative
period, besides providing adequate maintenance of the
vital signs during the clinical procedures, corroborating
with several authors [10,11,13]. The analysis of the variables such as age, surgical
time, cartidges number used, gender, systemic condition (ASA
classification), operated region (side, posterior or anterior,
upper or lower) showed procedure homogenization and
no interference with results, which is an important issue in
the present study [26]. The result of the patients’ anxiety
profile analysis demonstrated the predominance of anxious
and very anxious individuals, regardless of age and gender. RESULTS The center line represents the
median, the box represents the 1st and 3rd quartiles and the whiskers show the minimum and maximum values. Figure 5. Time spent (in seconds) during the completion of the Trieger test according to the groups and the verification periods. The center line represents the
median, the box represents the 1st and 3rd quartiles and the whiskers show the minimum and maximum values. Figure 5. Time spent (in seconds) during the completion of the Trieger test according to the groups and the verification periods. The center line represents the
median, the box represents the 1st and 3rd quartiles and the whiskers show the minimum and maximum values. RGO, Rev Gaúch Odontol. 2019;67:e20190060 AJB ROSA et al. Table 2. Volunteers’ perception on sedation induced by treatments. Table 2. Volunteers perception on sedation induced by treatments. Group 1 (oral)
Group 2 (sublingual)
P
Procedure recall
Everything
25 (96.2%)
23 (88.5%)
0.61000
Majority of the procedure
1 (3.8%)
3 (11.5%)
Effects reported
None
24 (92,3%)
14 (53,8%)
0.0001*
Muscle relaxation
1 (3.8%)
-
Drowsiness
-
11 (42.3%)
Drowsiness/Dizziness
-
1 (3,.%)
Dizziness
1 (3.8%)
-
Difference compared to previous procedure
Much
20 (76.9%)
21 (80.8%)
0.73**
Some
3 (11.5%)
3 (11.5%)
Little
2 (7.7%)
-
Indifferent
1 (3.8%)
2 (7.7%)
Note: *Comparison between none and drowsiness. **Comparison between “ much “ and all the other classifications together. Note: *Comparison between none and drowsiness. **Comparison between “ much “ and all the other classifications together. observed a predominance of anxious patients in several
dentistry specialties. RGO, Rev Gaúch Odontol. 2019;67:e20190060 REFERENCES 1. Candido MC, Andreatini R, Zielak JC, de Souza JF, Losso
EM. Assessment of anxiety in pa-tients who undergo
surgical procedures for tooth implants: a prospective study. Oral Maxillofac Surg. 2015 Sep;19(3):253-8. http://dx.doi. org/10.1007/s10006-014-0480-3 2. Crofts-Barnes NP, Brough E, Wilson KE, Beddis AJ,
Girdler NM. Anxiety and quality of life in phobic dental
patients. J Dent Res. 2010 Mar;89(3):302-6. http://dx.doi. org/10.1177/0022034509360189 Regarding the volunteers’ perception, they
reported presenting no undesirable side effects, and
most of them would choose to take alprazolam again if
they needed a new dental procedure. Furthermore, most
volunteers reported that they remembered the whole
procedure. Additionally, for the formulation of alprazolam
administered by the sublingual route, some of the
volunteers reported drowsiness, which can be explained by
the lower latency of the drug [30,31,34]. 3. Armfield JM. Predicting dental avoidance among dentally
fearful Australian adults. Eur J Oral Sci. 2013 Jun;121(3 Pt
2):240-6. http://dx.doi.org/10.1111/eos.12036 4. Carter AE, Carter G, Boschen M, AlShwaimi E, George
R. Pathways of fear and anxiety in dentistry: A review. World J Clin Cases. 2014 Nov; 2(11):642-53. http://dx.doi. org/10.12998/wjcc.v2.i11.642 5. Armfield JM, Stewart JF, Spencer AJ. The vicious cycle of
dental fear: exploring the interplay between oral health,
service utilization and dental fear. BMC Oral Health. 2007 Jan
14;7:1. http://dx.doi.org/10.1186/1472-6831-7-1 Although volunteers reported more drowsiness
with this formulation, the results of the Trieger test
demonstrate the patient’s rapid recovery, which suggests
the clinical feasibility of this formulation for Implantology
clinical procedures. It is also essential to highlight that no
evaluated formulation presented adverse reactions and
intercurrences, demonstrating that the clinical safety of this
type of benzodiazepine in clinical procedures [13,35]. Considering the profile of the patients evaluated, the
results obtained in the present study suggest that both
alprazolam formulations may be viable clinical options
for surgical procedures in Implantology. Considering
the importance of the subject and the scarcity of similar
studies, new studies should be carried out in order
to more demonstrate the use of alprazolam for surgical
Implantology procedures. 6. González-Lemonnier S, Bovaira-Forner M, Peñarrocha-Diago
M, Peñarrocha-Oltra D. Relationship between preoperative
anxiety and postoperative satisfaction in dental implant
surgery with intravenous conscious sedation. Med Oral Patol
Oral Cir Bucal. 2010 Mar 1;15(2):e379-82. http://dx.doi. org/10.4317/medoral.15.e379 7. Oosterink FM, de Jongh A, Aartman IH. What are people afraid
of during dental treatment? Anxiety-provoking capacity of 67
stimuli characteristic of the dental setting. Eur J Oral Sci. 2008
Feb;116(1):44-51. http://dx.doi.org/10.1111/j.1600-0722.20
07.00500.x 8. Collaborators AJB DA ROSA is the principal investigator and led the
writing of the manuscript. JC RAMACCIATO and RHL MOTTA are
the project managers, co-investigators and contributed to the
writing and revision of the manuscript. CE FRANCISCHONE, BS
SOTTO-MAIOR and RB BRITO JUNIOR are co-investigators and
contributed to the writing and revision of the manuscript. CONCLUSION It was concluded that both formulations of
alprazolam were effective for the sedation of the volunteers
evaluated. The Trieger test was chosen as assessment for
evaluating the effect of two routes of administration on
motor coordination, since it has been widely used and
recommended as a method of evaluating post-sedation
responsiveness for patient release [19,20]. The evaluation
of the results obtained by the Trieger test shows that the
motor coordination recovery was similar when the two
administration routes were compared. However, the time
taken to complete the test was significantly lower in the
final period for the two routes, indicative of increased
motor coordination of the patient due to the control /
reduction of preoperative anxiety. Thus, the results have
suggested that both formulations provide a rapid recovery
of the patient, probably due to their shorter half-life, which
s one of the reasons for their clinical indication for elderly
patients [11,13,21]. DISCUSSION This corroborates with different authors [7,27,29], who The use of the Corah scale chosen to evaluate
patients’ fear and anxiety is well publicized in the literature 8 Oral and sublingual sedation and its reliability and validity are supported by several studies
[16,17,33]. Moreover, visual analogue scale (VAS) has been
used in studies for many years and has been confirmed
as an effective method to evaluate the volunteers’ anxiety
[18]. 2010. Biomed Res Int. 2017;2017:2841549. http://dx.doi.
org/10.1155/2017/2841549 2010. Biomed Res Int. 2017;2017:2841549. http://dx.doi. org/10.1155/2017/2841549 24. Shavakhi A, Soleiman S, Gholamrezaei A, Khodadoostan M,
Shavakhi S, Tahery A, et al. Premedication with sublingual
or oral alprazolam in adults undergoing diagnostic upper
gastrointestinal endoscopy. Endoscopy. 2014;46(8):633-9. http://dx.doi.org/10.1055/s-0034-1377305 10. Corcuera-Flores JR, Silvestre-Rangil J, Cutando-Soriano A,
López-Jiménez J. Current methods of sedation in dental
patients - a systematic review of the literature. Med Oral
Patol Oral Cir Bucal. 2016 Sep 1;21(5):e579-86. http://dx.doi. org/10.4317/medoral.20981 25. Woolgrove J, Cumberbatch G. Dental anxiety and regularity
of dental attendance. J Dent. 1986 Oct;14(5):209-13. http://
dx.doi.org/10.1016/0300-5712(86)90003-5 11. Cogo K, Bergamaschi CC, Yatsuda R, Volpato MC, Andrade
ED. Sedação consciente com benzodiazepínicos em odonto-
logia. Rev Odontol Univ São Paulo. 2006;18(2)181-8. 26. Vassend O. Anxiety, pain and discomfort associated with
dental treatment. Behav Res Ther. 1993 Sep;31(7):659-66. http://dx.doi.org/10.1016/0005-7967(93)90119-f 12. Kaplan GB, Greenblatt DJ, Ehrenberg BL, Goddard JE,
Harmatz JS, Shader RI. Single-dose pharmacokinetics and
pharmacodynamics of alprazolam in elderly and young
subjects. J Clin Pharmacol. 1998 Jan;38(1):14-21. http://dx.doi. org/10.1002/j.1552-4604.1998.tb04370.x 27. de Carvalho RW, Falcão PG, Campos GJ, Bastos Ade S, Pereira
JC, Pereira MA, et al. Anxiety regarding dental treatment:
prevalence and predictors among Brazilians. Cien Saude
Colet. 2012 Jul;17(7):1915-22. http://dx.doi.org/10.1590/S14
13-81232012000700031 13. Haas DA. Oral sedation in dental practice. Royal College of
Dental Surgeons of Ontario; 2015. 28. Newton JT, Buck DJ. Anxiety and pain measures in dentistry: a
guide to their quality and ap-plication. J Am Dent Assoc. 2000
Oct;131(10):1449-57. http://dx.doi.org/10.14219/jada.archive. 2000.0056 14. Scavone JM, Greenblatt DJ, Shader RI. Alprazolam kinetics
following sublingual and oral administration. J Clin
Psychopharmacol. 1987 Oct;7(5):332-4. http://dx.doi.org/10. 1097/00004714-198710000-00008 29. Dailey YM, Humphris GM, Lennon MA. The use of dental
anxiety questionnaires: a survey of a group of UK dental
practitioners. Br Dent J. 2001 Apr 28;190(8):450-3. http://dx.doi. org/10.1038/sj.bdj.4801000 15. Matear DW, Clarke D. Considerations for the use of oral
sedation in the institutionalized geriatric patient during dental
interventions: a review of the literature. Spec Care Dentist. 1999 Mar-Apr;19(2):56-63. http://dx.doi.org/10.1111/j.1754-45
05.1999.tb01369.x 30. Wolf DL, Desjardins PJ, Black PM, Francom SR, Mohanlal
RW, Fleishaker JC. Anticipatory anxiety in moderately to
highly-anxious oral surgery patients as a screening model for
anxiolytics: evaluation of alprazolam. J Clin Psychopharmacol. 2003 Feb;23(1):51-7. http://dx.doi.org/10.1097/00004714-2
00302000-00009 16. Corah NL. Development of a dental anxiety scale. J Dent Res. 1969 Jul-Aug;48(4):596. http://dx.doi.org/10.1177/002203
45690480041801 17. Corah NL, Gale EM, IIlig SJ. Assessment of a dental anxiety
scale. J Am Dent Assoc. 1978 Nov;97(5):816-9. http://dx.doi. org/10.14219/jada.archive.1978.0394 31. REFERENCES Eli I, Schwartz-Arad D, Bartal Y. Anxiety and ability to recognize
clinical information in dentis-try. J Dent Res. 2008 Jan;87(1):65-8. http://dx.doi.org/10.1177/154405910808700111 9. Lino PA, Martins MAP, Silva MESE, de Abreu MHNG. Anxiolytics,
Sedatives, and Hypnotics Prescribed by Dentists in Brazil in RGO, Rev Gaúch Odontol. 2019;67:e20190060 AJB ROSA et al. 2010. Biomed Res Int. 2017;2017:2841549. http://dx.doi.
org/10.1155/2017/2841549 Joshi S, Ansari AS, Mazumdar S, Ansari S. A comparative
study to assess the effect of oral alprazolam as premedication
on vital parameters of patients during surgical removal of
impacted mandibular third molars. Contemp Clin Dent. 2016
Oct-Dec;7(4):464-468. http://dx.doi.org/10.4103/0976-237X. 194125 18. Facco E, Zanette G, Favero L, Bacci C, Sivolella S, Cavallin
F, et al. Toward the validation of visual analogue scale for
anxiety. Anesth Prog. 2011 Spring;58(1):8-13. http://dx.doi. org/10.2344/0003-3006-58.1.8 32. Van Den Berg F, Tulen JH, Boomsma F, Noten JB, Moleman
P, Pepplinkhuizen L. Effects of alprazolam and lorazepam
on catecholaminergic and cardiovascular activity during
supine rest, mental load and orthostatic challenge. Psychopharmacology (Berl). 1996 Nov;128(1):21-30. http://
dx.doi.org/10.1007/s002130050105 19. Trieger N, Newman MG, Miller JC. An objective measure of
recovery. Anesth Prog. 1969 Jan;16(1):4-7. 20. Newman MG, Trieger N, Miller JC. Measuring recovery from
anesthesia-a simple test. Anesth Analg. 1969;48(1):136-40. 21. Andrade ED. Terapeutica Medicamentosa na Odontologia. 3ª
ed. São Paulo: Artes Médicas; 2014. 33. Sharma A, Pant R, Priyadarshi S, Agarwal N, Tripathi S,
Chaudhary M. Cardiovascular Changes Due to Dental Anxiety
During Local Anesthesia Injection for Extraction. J Maxillofac
Oral Surg. 2019 Mar;18(1):80-87. http://dx.doi.org/10.1007/s12
663-018-1085-4 22. Resnik RR, Misch C. Prophylactic antibiotic regimens in oral
implantology: rationale and protocol. Implant Dent. 2008
Jun;17(2):142-50. http://dx.doi.org/10.1097/ID.0b013e3181
752b09 34. Standards for Conscious Sedation in the Provision of Dental
Care. The dental faculties of the royal colleges of surgeons
and the Royal College of Anaesthetists. 2015. 23. Berthold CW, Dionne RA, Corey SE. Comparison of sublingually
and orally administered tria-zolam for premedication before
oral surgery. Oral Surg Oral Med Oral Pathol Oral Radiol
Endod. 1997 Aug;84(2):119-24. http://dx.doi.org/101016/
s1079-2104(97)90055-x 35. Donaldson M, Gizzarelli G, Chanpong B. Oral sedation:
a primer on anxiolysis for the adult patient. Anesth Prog. 2007;54(3):118-28. http://dx.doi.org/10.2344/0003-3006 Received on: 15/2/2019
Final version resubmitted on: 8/5/2019
Approved on: 20/5/2019 Received on: 15/2/2019
Final version resubmitted on: 8/5/2019
Approved on: 20/5/2019 10 RGO, Rev Gaúch Odontol. 2019;67:e20190060 10
|
https://openalex.org/W4310433449
|
https://jovacet.ac.za/index.php/JOVACET/article/download/250/133
|
English
| null |
TVET leaders’ experiences of an enquiry-based blended-learning programme
|
Journal of vocational, adult and continuing education and training
| 2,022
|
cc-by
| 8,303
|
TVET leaders’ experiences
of an enquiry-based
blended-learning programme Tanya Smit and Suzanne Bester
University of Pretoria KEYWORDS
Vocational education, TVET, leadership development, blended learning, enquiry-based learning Background and context Technical and vocational education and training colleges (TVET) in South Africa have been
in existence for 20 years after the former technical colleges and colleges of education and
training centres were merged in 2002. Considerable expectations existed about how these 50
colleges would become more vital institutions with the capacity to teach increased numbers
of students in a variety of programmes (HRDC, 2013). Despite having received substantial
funding from the government at the launch of the TVET colleges, these institutions still face
significant challenges (Mohlokoane & Coetzer, 2007; Badenhorst & Radile, 2018). In
assessing the quality of TVET colleges, Worku (2019) identified a lack of leadership as one
of the primary challenges. This lack of leadership was identified in both administrators and
academics, which resulted in a lack of resources, poor curriculum design, a lack of quality
lecturers and inadequate facilities in colleges (Mohlokoane & Coetzer, 2007; Badenhorst &
Radile, 2018; Robertson & Frick, 2018; Worku, 2019). Specific areas of underperformance
include a failure to attend scheduled council board meetings, to form partnerships with
relevant stakeholders or to ensure that the curricula are aligned to the labour markets (Worku,
2019). It was also found that more accountability, fairness, objectivity and transparency were
needed to warrant the effective use of college resources (Worku, 2019). In a study by Robertson and Frick (2018), it was found that TVET managers often come straight
from a teaching environment and are expected to lead and manage TVET colleges without having
undergone the necessary training. Furthermore, the studies by Worku (2019) and Robertson and
Frick (2018) show the gap between what is expected of TVET managers and their current
performance, indicating a need for them to participate in further training and development
(Worku, 2019). Similarly, Robertson and Frick (2018) alluded to the need for TVET managers to
obtain the necessary skills and knowledge to manage and lead their institutions effectively in order
to meet the mandate set for TVET colleges. But investment in the professional development of
TVET managers is needed to enable the quality of leadership in the colleges to improve. Improving
leadership could, in turn, result in improved curriculum design and implementation, which could
lead to the quality graduates needed in the labour market being produced (Balkrishen & Mestry,
2016; Badenhorst & Radile, 2018; Worku, 2019). ABSTRACT A grave need exists among managers in Technical and Vocational Education and Training (TVET) in
South Africa to have their leadership and managerial skills developed. The Postgraduate Diploma
in TVET programme at the University of Pretoria was developed in 2019 as an enquiry-based
blended-learning programme aimed at contributing to an appropriate response to this need. The
purpose of this exploratory case study was to describe the experiences of the first cohort of
programme participants according to the way the design of this programme influenced their
engagement in the programme and how it supported the development of their leadership and
managerial skills. Data were generated using reflective essays and a group reflection session with
all the participants. The essays and the transcription of the group reflection session were
thematically analysed. The findings suggest that the programme promoted the development of
the participants’ leadership and managerial skills. It also developed their critical thinking skills,
their ability to work independently and their inclination to become self-efficacious learners. The
facilitators in this programme became powerful role models to the research participants to
replicate their learning experiences in their own institutions. Although the participants found it
challenging to adjust to working from home while also studying during the COVID-19 lockdown
period, they were inspired and motivated to persist through their networking experiences with
international and national peers and leaders in their fields of education and industry. KEYWORDS
Vocational education, TVET, leadership development, blended learning, enquiry-based learning — 136 — TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester Background and context A needs analysis conducted in 2018 by the Department of Higher Education and Training
(DHET, 2018) similarly identified the skills gap evident in TVET college leaders. The Post
Graduate Diploma in Technical and Vocational Education and Training (PGDip in TVET)
programme that is reported on in this article was developed at the request of the DHET in
support of the College Lecturer Education Project which was part of the Teaching and
Learning Development Capacity Improvement Programme (TLDCIP) – a partnership
programme between the DHET and the European Union (EU). The University of Pretoria, in consultation with TVET colleges, the DHET, industry and the
Technical University of Munich (TUM), developed the PGDip TVET programme in 2019. The German Agency for International Cooperation (GIZ) funded the development of the — 137 — Journal of Vocational, Adult and Continuing Education and Training 5(1) 10.14426/jovacet.v5i1.250 programme and the Education, Training and Development Practices Sector Education and
Training Authority (ETDP SETA) sponsored the tuition fees of the programme’s participants. This internationally benchmarked, one-year enquiry-based blended-learning programme was
envisaged as a means of supporting leaders in the TVET sector in developing strategic
leadership skills and advancing their professional competency. The programme consists of eight compulsory modules and one optional module. The
purpose of the programme, as illustrated in Figure 1, is to create enabling opportunities for
TVET leaders that could help them to develop the professional knowledge, skills and values
required to become innovative, visionary and strategic leaders who are able to navigate
context-specific demands and perform leadership functions in the following key performance
areas (KPAs): Curriculum Design and Development, Managing and Leading People,
Managing physical infrastructure and Finance with the last about developing innovation and
quality. The professional development and mentoring framework was central to the
programme and it provided opportunities for the participants to develop and network with
other TVET leaders or experts. Various approaches to enquiry-based learning – such as
problem-based learning, project-based learning, game-based learning and discovery learning
– were used in the different modules. The programme leaders took a decision that the word
‘student’ would not be used when referring to the managers and leaders who decided to enrol
for the programme. Instead, they would be called ‘programme participants’. Background and context The reason for
this is that the teaching and learning approach in the programme was collaborative and
participatory in nature and therefore it made sense to refer to the future graduates in the
programme as programme participants. Figure 1 shows the structure of the PGDip in TVET programme and how all eight modules,
the KPAs and the mentoring framework are interlinked. The programme challenges the
participants to understand the TVET context, think critically, communicate, collaborate,
and develop their professional and strategic leadership competencies as TVET leaders. The Professional Development (PFO 700) module is an integration of all the modules and
KPAs. In KPA 1, the modules Curriculum Leadership (CDD 711) and Instructional Leadership
(LMD 700) are included. The modules Emotional-Social Well-being (KGG 731) and
Management and Leadership in Education (LVO 732) form part of KPA 2. In KPA 3, the
Financial and Human Resource Management (EDM 734) module is a compulsory module and
then the research module NMQ 745 and 755 are optional modules that programme participants
could take in addition to the eight compulsory modules if they wish to pursue postgraduate
studies further. In KPA 4 the modules included are Educational Technology in Higher
Education (OWT 730) and Assessment and Quality Assurance (QPI 713). Following the implementation of this programme in 2020, the programme developers and
facilitators embarked on a series of research studies to investigate various aspects of this
programme. This study reports on one aspect currently under investigation: the experiences
of the first cohort of participants in the enquiry-based blended-learning programme. — 138 — TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester p
f
q
y
g p g
FIGURE 1: Overview of the PGDip in TVET programme and module alignment
Mentoring
Framework
KPA2
KPA1
LMD
700
EDM
734
KGG
731
QPI
713
CDD
711
MNQ
745 and
755
LVO
732
OWT
730
KPA4
KPA3
Peer mentoring
Peer mentoring
Peer mentoring
Peer mentoring FIGURE 1: Overview of the PGDip in TVET programme and module alignment Conceptual framework The fundamental changes in the global economy, jobs and the industry have reshaped the
nature of work, driving new and different skills compared to those required in the past
(Choudaha & Van Rest, 2018). According to the World Economic Forum’s (WEF, 2020) ‘the
world of of jobs’ report, the top two skills that will grow in prominence in the next five years
are critical-thinking and problem-solving skills. Newly emerging skills this year are those in
self-management such as active learning, resilience, stress tolerance and flexibility – which is
why, according to Gleason (2018), higher education needs to prepare thinkers of the Fourth
Industrial Revolution (4IR) more adequately. These changes in the world of work drive — 139 — Journal of Vocational, Adult and Continuing Education and Training 5(1) 10.14426/jovacet.v5i1.250 changes in educational practices, and for this reason authors such as Jonker, März and Voogt
(2018) and Lubua (2019) suggest flexible, diverse and individualised approaches to learning. According to Chu, Reynolds, Tavares, Notari and Lee (2017), enquiry-based approaches to
learning are vital for developing 21st-century skills and practices. Simply stated, enquiry-based learning emphasises an active approach to learning, which
assumes that learners are actively involved in their learning, solving problems unique to
their contexts and constructing new knowledge in the process. Pedaste, Mäeots, Siiman,
De Jong, Van Riesen, Kamp, Manoli, Zacharia and Tsourlidaki (2015) cite several studies
that provide evidence to support the effectiveness of enquiry-based learning and state that
it is generally regarded as a vital element of building a scientifically literate community. As
part of an enquiry-based approach to learning, learners are normally guided through an
authentic process of scientific discovery. They are encouraged to formulate problems, ask
questions, construct knowledge, formulate solutions and then evaluate, refine and test
their solutions. It is important to remember that enquiry-based learning is not a linear
process and that it requires active engagement in a continuous cycle of exploration and
refinement. Facilitating learning in accordance with this approach implies that lecturers or
facilitators play a facilitating and mediating role in learning and that no ‘traditional’
teaching practice is involved. Technical education is extremely important to the development of an industrial society,
but in the post-industrial world employers value softer skills such as leadership skills,
creativity, the ability to think outside the box and openness to multiple perspectives. Conceptual framework These
skills are poised to become more important as artificial intelligence replaces human workers
in many technical fields (Lewis, 2018). According to Lewis (2018), higher education needs
to emphasise pedagogy that is student-centred and individualised. Assessments are arguably
the most effective when grounded in project-based learning and authentic experiences. In
the enquiry-based learning approach adopted as a conceptual framework for this
programme, the programme participants followed methods and practices similar to those
of professional scientists in order to construct knowledge (Pedaste et al., 2015) and they
were mentored to move through five distinct phases of enquiry: orientation,
conceptualisation, investigation, conclusion and discussion. The cycle is often viewed as an
approach to solving problems and involves the application of several problem-solving
skills. The findings on the merits of enquiry-based approaches to learning are mixed, though. Whereas some consider it to be an effective instructional approach or educational strategy
(Alfieri, Brooks, Aldrich & Tenenbaum, 2011; Furtak, Seidel, Iverson & Briggs, 2012),
others suggest that it may not offer any special advantages over traditional approaches to
teaching and learning (Jerrim, Oliver & Sims, 2019). It has also been noted that the
implementation of enquiry-based learning creates challenges such as a lack of resources and
training, an over-emphasis on assessing content, the level of difficulty of this approach and
the time-consuming nature of enquiry-based learning (Gutierez, 2015). — 140 — TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester The enquiry-based approach followed in this programme has two elements that the authors
hypothesised could strengthen the effects of such learning on student achievement. First, it
is designed around a three-pillar mentoring framework (individual, peer and expert
mentoring) and, second, the online blended-learning component has been designed to
prepare and engage programme participants maximally through online learning activities
(such as podcasts, videos) and professional self-assessment activities (leadership competencies
and skills, readiness for learning and engaging). The learning approach of the PGDip in
TVET programme aligns with Cronje’s definition of blended learning, which the author has
described as ‘the appropriate use of a mix of theories, methods and technologies to optimise
learning in a given context’ (Cronje, 2020:120). Conceptual framework In alignment with the enquiry-based learning approach, the programme participants were
given opportunities to think critically, solve real-life problems and develop technological
literacy while engaging in instructional leadership design and development. In addition, the
blended-learning meta-structure used in the PGDip in TVET programme emphasised active
engagement: the programme participants were challenged to think, plan and make decisions
strategically so as to be agents of change and to emerge as visionary leaders. Pedaste et al. (2015) agree on the effectiveness of enquiry-based learning since it focuses on
the student and is regarded as a vital element in building a scholarly community. The TVET
leadership programme participants took responsibility for their learning while engaging and
participating in the four contact sessions, continuous blended-learning activities and a study
visit to Germany. The objectives of the study visit component were for the programme
participants to engage with experts in the dual approach to vocational education in Germany
and to collaborate on strategic plans that they could implement in the South African context. Unfortunately, the study visit had to be adjusted due to the COVID-19 pandemic: instead of
an in-person visit, the programme participants engaged in streaming video tours of German
industry and TVET college sites, using recorded or live interviews, webinars and presentations
at study visit events or seminars. The programme participants also took part in collaborative
sessions on Design Thinking. This contextually driven change to the programme was
supported by Zhao and Watterston’s (2021) view that COVID-19 constructed a distinctive
opportunity for education change. As stated previously, the programme’s components were integrated and employed a three-
pillar mentoring approach, namely individual professional mentoring, peer-group mentoring
and KPA mentoring. Various components, activities, media, technologies and support
materials were included. The meta-structure of the programme expected the participants to
participate actively during, for instance, the action research, expert panel discussions, podcast
discussions, reflective journal writing, the design of posters, scholarly readings and the
making of videos and presentations. Instructional technology and apps such as Linoit,
Mentimeter, Miro, Wiki’s, Loom, Anchor, PhotoCircle and Canva were used. The programme
participants engaged continuously with their peers and national, international, industry and
higher education experts or facilitators. — 141 — Journal of Vocational, Adult and Continuing Education and Training 5(1) 10.14426/jovacet.v5i1.250 Journal of Vocational, Adult and Continuing Education and Training 5(1) 10.14426/jovacet.v5i1.250 The participants resided across South Africa and had demanding work schedules as TVET
leaders. Conceptual framework Apart from their busy schedules, they entered this programme at a time when
struggles to access electricity, stable internet data, learning materials, etc, were commonplace
for most learners in South Africa. Cronje (2020:117) cautions that there must be ‘sensitivity
to the context in which learning occurs’ for blended learning to be successful. During the programme and following the enquiry-based blended-learning approach, the
participants progressed through a non-linear scholarly discovery process by formulating
problems, asking questions, constructing knowledge and identifying, implementing,
evaluating, refining and testing solutions. Dewey agrees that enquiry is ‘a backward and
forward connection, a discovery of the connection of things’ (Dewey, 1916:206). The PGDip
in TVET enquiry-based framework was adapted from Pedaste et al. (2015) and implemented
as outlined in Figure 2. ORIENTATION
CONTACT 1
PROGRAMME MODULES
BLENDED-ONLINE-LEARNING-COMPONENTS
CONCEPTUALISATION
CONTACT 2 AND 3
INVESTIGATION
CONCLUSION
CONTACT 4
FIGURE 2: PGDip in TVET adapted an enquiry-based blended-learning framework
CONCLUSION
DATA-INTERPRETATION
ORIENTATION
HYPOTHESIS-GENERATION
EXPERIMENTATION
QUESTIONING
EXPLORATION PROGRAMME MODULES CONTACT 2 AND 3 FIGURE 2: PGDip in TVET adapted an enquiry-based blended-learning framework — 142 — TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester The programme participants progressed through four stages of the framework, as illustrated
in Figure 2. During the orientation phase, the participants explored a problem regarding
their strategic plan development. They reflected critically on their TVET college and
stimulated curiosity and interest in their chosen problem as a means of exploring their
strategic plan development. The second phase, conceptualisation, entailed creating theory-based questions or hypotheses. This phase was divided into two sub-phases: questioning and hypothesis generation. The
programme participants questioned and tried to understand the concepts connected to their
identified problem. During questioning, they formulated an open question about their
context. In addition, they formulated a vision for their TVET college. In the third phase, investigation, further exploration was planned, and the data or
information was collected and analysed. In the last phase of the framework process, the
programme participants completed their strategic plan by drawing conclusions from the
data. The overarching communication phase was an external but integrated part of the
enquiry-based blended-learning process, as it was present in every phase. Throughout, the
programme participants communicated and gained feedback from their peers and mentors
to help articulate their understanding. Conceptual framework Furthermore, they reflected on their learning as
Leijen, Allas, Toom, Husu, Marcos, Meijer, Knezic, Pedaste and Krull (2014) advised by
describing, justifying, critiquing and discussing their experiences, their learning and the
meaning made. In the third phase, investigation, further exploration was planned, and the data or
information was collected and analysed. In the last phase of the framework process, the
programme participants completed their strategic plan by drawing conclusions from the
data. The overarching communication phase was an external but integrated part of the
enquiry-based blended-learning process, as it was present in every phase. Throughout, the
programme participants communicated and gained feedback from their peers and mentors
to help articulate their understanding. Furthermore, they reflected on their learning as
Leijen, Allas, Toom, Husu, Marcos, Meijer, Knezic, Pedaste and Krull (2014) advised by
describing, justifying, critiquing and discussing their experiences, their learning and the
meaning made. Research theory underpinning this study For this study, an interpretivist meta-theoretical paradigm and a qualitative methodological
paradigm were used. Interpretivists believe that people construct reality through their lived
experiences and that one needs to consider the context in which research is conducted in
order to interpret data accurately (Willis, 2007). Interpretivists value subjectivity and believe
that reality is constructed by means of individual interpretations through which people
construct their own meaning of events (Mack, 2010). Interpretivists view events as distinctive
phenomena that cannot be generalised and include the acceptance of multiple perspectives
on and experiences of one incident (Mack, 2010). Therefore, researchers within this paradigm
explore real-world situations that unfold naturally while they remain unobtrusive, non-
controlling and non-manipulative observers (Tuli, 2010). Qualitative approaches are generally preferred methods of enquiry within an interpretivist
paradigm (Thomas, 2003; Willis, 2007). Such approaches often yield the rich information
necessary to understanding specific contexts accurately (Willis, 2007). Qualitative research is
an exploratory approach to enquiry that aims to understand the meaning that individuals or
groups assign to social phenomena (Creswell, 2014). — 143 — Journal of Vocational, Adult and Continuing Education and Training 5(1) 10.14426/jovacet.v5i1.250 Similarly, in this study, the authors believed that the participants had varying real-life
experiences that could influence their viewpoints about the way in which this programme
influenced their engagement and how it supported the development of their leadership and
managerial skills. Since the participants were directly exposed to the programme, the authors
believed that their personal viewpoints would inform the purpose of this study. Through
qualitative enquiry, they were able to ask broad, open-ended questions that enabled the
participants to respond freely in reflecting on their experiences of the programme. By
exploring those experiences, the authors could reflect on the implications for further research
and for enhancing the programme. Purpose of the study The purpose of this interpretivist, qualitative exploratory case study was to describe the
experiences of the first cohort of participants to enrol for the PGDip in TVET programme
in 2020, relating to the way in which the design of this programme influenced their
engagement in the programme and how it supported the development of their leadership and
managerial skills. Although this study is not aimed specifically at programme design
improvement, it could offer insight into aspects of the programme that might need refinement
and improvement based on the participants’ experiences. It could also inform other
programme developers who want to develop similar programmes in the future. Participants and recruitment The project received ethical clearance from the Research Ethics Committee in the Faculty of
Education. The participants were recruited during the first contact session in February 2020
before the programme commenced. The recruitment was done during a face-to-face information
session before the COVID-19 lockdown came into effect. The participants received verbal and
written information about the purpose of the study and the data that could be included in it. Their participation was motivated by the altruistic notion that their contribution could
influence the improvement of the programme, benefitting those who would be following in
their footsteps. All 29 participants opted to be included as research participants in the study. They were also informed that non-participation would not affect them academically and that
they could withdraw at any time by sending an email to the programme coordinator. This was
reiterated before every data-collection activity and again at the end of the programme. Drawn from TVET colleges located around the country, most of the participants (51%) were
men. One participant had a PhD and two had previously obtained Master’s degrees in
education. Of the participants, 12 held Honours degrees, 12 had obtained BEd degrees and
two had BCom degrees. Twelve of the participants were campus managers, six were deputy
principals, three were heads of school, three were heads of department and one was a TVET
college principal. The participants were all older than 40, with half being over 50 and three
being older than 60. — 144 — TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester Data collection and analysis Data were generated using reflective essays and an online group reflection. The participants
were asked to reflect on the following topics: •
How the programme dealt with their needs or expectations as TVET managers (or
not). •
How they experienced the design of the programme. •
Whether they preferred the blended approach instead of a traditional face-to-face
only approach, and why. The reflective essays, the recording and the transcription of the group reflection session were
analysed thematically using a six-phase reflexive thematic framework (Braun & Clarke, 2020). The data were analysed inductively. Initial categories were generated, analysed for similarities
and collated into preliminary codes that identified major themes. Findings Two major themes emerged in the data. The first related to the perceived benefits or gains the
participants reported, whereas the second related to the challenges they faced or how the
programme had not met their needs. The two themes are discussed under the sub-themes
developing leadership and managerial skills, the blended-learning approach, and the enquiry-
based approach. Theme 1: Benefits and gains Developing leadership and managerial skills Developing leadership and managerial skills The TVET programme places a strong emphasis on networking and mentoring as leadership
skills and the participants in the programme were exposed to various opportunities to
experience the value of mentoring and networking. Concerning their networking experiences
in the programme, one of the participants stated: [W]e always worked in isolation as TVETs; this programme has actually brought
us close to even share knowledge and resources outside this programme. Another felt that the networking opportunities reframed the way he viewed himself and his
college, stating: Another felt that the networking opportunities reframed the way he viewed himself and his
college, stating: … having to see myself and my college as part of a global village is quite great. … having to see myself and my college as part of a global village is quite great. is sentiment was shared by another participant, who stated: This sentiment was shared by another participant, who stated: This sentiment was shared by another participant, who stated: I am now thinking outside my environment – looking at provincial, national,
continental and global perspectives. — 145 — Journal of Vocational, Adult and Continuing Education and Training 5(1) 10.14426/jovacet.v5i1.250 The programme encouraged the participants to evaluate not only their own leadership and
managerial practices but also the practices that exist in their colleges. One participant stated: The programme encouraged the participants to evaluate not only their own leadership and
managerial practices but also the practices that exist in their colleges. One participant stated: [I]t [the programme] helped me take an in-depth look at my leadership strengths
and weaknesses. Several participants commented on how the programme had helped them to develop specific
skills, such as strategic planning. One participant concluded: Several participants commented on how the programme had helped them to develop specific
skills, such as strategic planning. One participant concluded: The college has year after year developed a strategic plan but consistently failed to
achieve the set objectives. I now understand why we failed to achieve our
objectives. We only developed the plan but ignored the organising, delegating,
and coordination of leadership and management tasks. Maybe it might be that we
lacked knowledge on those concepts. The same participant also concluded by remarking: [I]t is now up to me to be the change agent in the college by ensuring that the
knowledge gained in this course is put to good use and also benefit[s] the college. Developing leadership and managerial skills Some participants also reported developing soft skills such as being more inclusive and
accepting of others, gaining more confidence, managing conflict better, involving staff, and
focusing on improving staff and student morale and well-being. One participant said the
following in this regard: I became more consultative and inclusive in decision-making processes because of
this programme. I became more consultative and inclusive in decision-making processes because of
this programme. Blended learning As stated earlier, the blended learning in this programme consisted of various activities such as
self-study, podcasts, videos, assignments, mentoring, discussions, face-to-face contact sessions
and a study visit to industry partners. When asked if they preferred a blended-learning approach
to the traditional face-to-face only approach, the participants indicated that they preferred the
blended learning. They believed the online activities augmented the face-to-face virtual contact
sessions during block weeks. The following two quotations from the participants illustrate this: E-learning allows more effective interactions between me and my instructors with
emails, discussion boards and chat rooms. Blended learning enabled me to access
the materials from anywhere at any time while enjoying the benefits of face-to-
face support and instruction through contact sessions. [P]rogramme participants from various provinces in our country could interact
and engage meaningfully with learning without having to be in the same physical — 146 — TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester room with the facilitators. Hybrid technology successfully replaced the physical
classroom. room with the facilitators. Hybrid technology successfully replaced the physical
classroom. The University of Pretoria, where this programme is presented, is a contact learning
institution and the initial planning for the programme was that the face-to-face contact
sessions would be conducted in person on campus. Similarly, the scholarly visit to TUM in
Germany would have been in person. However, this plan had to be amended due to the hard
lockdown that existed in South Africa and globally for most of 2020, when this programme
was launched. Both the face-to-face contact sessions and the study visit had to be moved
online, with no direct in-person contact taking place between the facilitators and the
programme participants. Although the participants expressed their gratitude that the
programme could continue online, the message was also clear that they needed both online
activities and in-person contact with their facilitators and the peer group. This is evident in
the following statement by one of the participants: Although we could continue studying online, I was so much looking forward to
experience having contact sessions at the university. This was confirmed by another participant, who stated that they needed the human contact
associated with face-to-face learning. Another benefit that the participants reported was that the programme enabled mastery
learning. They were able to learn at their own pace, in their own time, through a variety of
activities. This is evident from the following quotations: I can tailor my learning experience while at the same time offering flexible time frames
that can be personalised to me, offering me the ability to learn at my own pace. I can tailor my learning experience while at the same time offering flexible time frames
that can be personalised to me, offering me the ability to learn at my own pace. Learning platforms provide access to all kinds of multimedia and interactive
content that significantly expand classroom face-to-face interaction. It was more engaging to me and enabled me to manage time efficiently. Kept me
to plan and to develop self-accountability while at the same time learning skills in
the use of technological devices/equipment such as computers to study. It was more engaging to me and enabled me to manage time efficiently. Kept me
to plan and to develop self-accountability while at the same time learning skills in
the use of technological devices/equipment such as computers to study. room with the facilitators. Hybrid technology successfully replaced the physical
classroom. A third benefit was that the participants believed the programme enabled them to take
responsibility for their own learning and to master various skills. This is evident from the
following statement: It challenges programme participants on how we do things in going forward as it
is bringing a different dimension on how a student conducts his or her studies. The online component of this programme had a very positive impact on my
studies and has taught me self-reliance and discipline skills. — 147 — Journal of Vocational, Adult and Continuing Education and Training 5(1) 10.14426/jovacet.v5i1.250 Finally, the programme inspired the participants, who are teachers themselves, to use their
blended-learning experiences to transform their own colleges. At the time of this study, they
were struggling with the implementation of online learning that had been forced on them by
the COVID-19 lockdown regulations. This is evident in the following two statements: The impact is so great to the extent that my college has started doing the same. I
made inputs based on what I was exposed to in this programme and now for me
teaching and learning has no borders. … as a campus manager leading one of the campuses in the deep rural province
of the Eastern Cape with poor facilities and infrastructure not conducive for
virtual teaching and learning, noting that most students own smart phones, I
inspired lecturers to make sure they interact with students by all means. Enquiry-based learning The enquiry-based learning approach that was followed in this programme aimed to
emphasise the participants’ role in the learning process and to encourage them to explore, ask
questions, share ideas and promote their critical thinking skills. Some of the participants
commented on the ways in which the programme had encouraged them towards self-efficacy
and to apply what they had gained from the programme in their own colleges. The following
quotations illustrate this: It [the programme] encourages independence and innovation. The programme is very well suited to a business development manager who must
at all times encourage innovation and creativity in the college offering. It [the learning approach of the programme] is the most creative way of producing
learners who will be able to research and look for information whenever they are
in a particular situation. Hybrid learning [with reference to the entire approach in the programme] is
important because it breaks down the traditional walls of learning, and now with
access to present-day technologies and resources hybrid learning forces time
management, working independently, flexibility in your learning as [a] student
and has taught me independence and discipline. Personally, it has covered all the
critical skills needed by a leader in an ideal TVET college. Through this programme I have learned that for every challenge there is a
solution. Through enquiry-based learning one can be able to understand the
problem [one is] faced with, identify the reasons why the problem exists and
explore possible solutions available and decide on the best solution to address the
problem. — 148 — TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester Theme 2: Challenges and unmet needs — 149 — Journal of Vocational, Adult and Continuing Education and Training 5(1) 10.14426/jovacet.v5i1.250 Journal of Vocational, Adult and Continuing Education and Training 5(1) 10.14426/jovacet.v5i1.250 The programme did not address the capacity to deal with strikes by students
and staff and the political tension that mobilise[s] and cause[s] college
instability. Blended learning The challenges the participants experienced with the learning activities primarily related to
practical matters such as having limited or no access to the internet or experiencing problems
with connectivity. An unreliable electricity supply and having to balance their work lives with
learning activities were challenging. The participants dealt with many upheavals in their schedules and faced staff challenges
arising from COVID-19. This is evident in the following statements: The important ingredient [in the programme] is the access to a stable Wi-Fi and
reliable electricity connections. The important ingredient [in the programme] is the access to a stable Wi-Fi and
reliable electricity connections. Online interaction was possible though not as convenient at times (network
problems, wrong times, and congested college). Online interaction was possible though not as convenient at times (network
problems, wrong times, and congested college). Theme 2: Challenges and unmet needs The benefits and gains the participants in this study reported exceeded the challenges and
unmet needs they shared. The researchers reflected on why this was the case and concluded
that there may have been many explanations. For instance, it could be that they mostly had
overwhelmingly positive experiences of the programme. Alternatively, it could be that they
wanted to please the facilitators, who were also the data collectors, or that they felt they could
not criticise the programme because they were sponsored financially by the DHET/ETDP
SETA and had been selected above other candidates to be part of this programme. The
researchers comment on this again in the ‘Limitations’ section below. Developing leadership and managerial skills Three participants reported that COVID-19 posed a significant challenge to acquiring all the
necessary leadership skills. One participant believed the pandemic, with the associated
lockdown regulations, to a certain extent limited his opportunities for networking and
receiving mentoring. This is illustrated by the following statement: COVID-19 did have an impact on my progress in the PGDip programme because
we could not visit Germany to be fully exposed to the dual approach system for
vocational education. A second participant noted that her progress was affected by the many changes she had to
adapt to which affected her progress, such as: A second participant noted that her progress was affected by the many changes she had to
adapt to which affected her progress, such as: [A]djusting to work from home, attending online meetings, at times managing
the crisis of infected colleagues and family. A third participant indicated that: [T]he pandemic increased my workload at work and that impacted my studies
negatively … and I felt overwhelmed at some point. Some expectations of the programme regarding the development of specific leadership and
managerial skills that were not met are evident in the following statements: Project management was not addressed. As leaders we are often expected to
manage projects. I wish the strategic plan was aligned to monitoring and evaluation of the action
plans developed. I wish the strategic plan was aligned to monitoring and evaluation of the action
plans developed. The programme is more focussed on teaching and learning and it does not talk of
support staff of which (sic) at the college they play an important role. One participant highlighted various issues they struggled with in the programme: It [the programme] was difficult for an older learner like myself who had to make
an extra effort … I felt sort of out of my depth and was shy to ask questions in
front of my peers. Enquiry-based learning When they developed this programme, the devlopers were concerned about the complexity
of enquiry-based learning. They felt this could be a potential barrier to the participants’
learning if they had demanding work schedules and, in most instances, they had not
undertaken postgraduate studies at all or recently. Some of the participants commented on
how they initially felt overwhelmed by the programme and struggled to find some rhythm
and routine. One participant stated that she felt overwhelmed by the programme’s complexity
during the orientation session, where they were introduced to the programme structure and
the mode of delivery. She stated the following: During orientation the programme seemed complicated and I was not sure if it
was a good decision for me to continue. A few participants commented on how they struggled to adjust to the programme. One
stated: It was a struggle at first. Another stated their challenges a little differently: At the beginning of the course, I was unable to balance my work and my studies
but that changed after developing a strategy to at least, in a day, read or complete
an activity; that is when I started to enjoy the online learning and sharing
information with some of my colleagues. — 150 — TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester Discussion The findings of this study show that the subjective experiences of the participants of the
enquiry-based blended-learning design of the PGDip in TVET programme were
predominantly positive. The programme encouraged the participants to evaluate themselves
critically as leaders and managers and also to evaluate the leadership practices at their
colleges. It challenged them to identify the problems and challenges they encountered in
their colleges and to develop a strategic vision and plan to resolve them. The programme
also promoted the development of soft skills such as being flexible and adapting to change,
being resilient and overcoming challenges, and being inclusive in decision-making
processes. This finding is significant when one considers that this programme is an attempt to redress
the lack of leadership in TVET colleges, as identified by various scholars, with a call for
increased accountability and objectivity in colleges (Mohlokoane & Coetzer, 2007;
Badenhorst & Radile, 2018; Robertson & Frick, 2018; Worku, 2019). Our findings concerning the enquiry-based and blended-learning approach followed in the
programme show that these approaches to learning complement each other. The combined
approaches encouraged the participants to become independent, critical and self-efficacious
learners. They discovered that co-constructing their learning with peers and experts in the
TVET sector creates a strong community of practice that promotes their learning even
during challenging times. These findings have implications for practice. They align with Jonker, März and Voogt
(2018) and Lubua (2019), who recommend flexible, diverse and individualised approaches
to learning which prepare learners for the 4IR. This can be achieved by developing skills
such as critical thinking, problem-solving and self-management, including active learning,
resilience, stress tolerance and flexibility (WEF, 2020). The challenges the participants shared were driven mainly by the adverse circumstances
that they faced as a result of the COVID-19 pandemic. This also affected the way in
which the programme was delivered. Similarly, access to resources such as electricity and
stable internet connectivity played a significant role in the way the participants
experienced this programme – which relied heavily on these resources. Discussion Given that for
learners in South Africa these challenges are unlikely to disappear in the foreseeable
future, it is essential that TVET facilitators who plan to use enquiry-based blended- — 151 — Journal of Vocational, Adult and Continuing Education and Training 5(1) 10.14426/jovacet.v5i1.250 learning methods are sensitive to these challenges and plan to overcome and manage
them in ways that will not compromise learning (Cronje, 2020). It is also important to
note that when challenging pedagogical approaches such as enquiry-based learning are
pursued, learners may feel overwhelmed and this may inhibit their growth and undermine
their self-belief. It may be necessary, therefore, to normalise the challenging nature of
this approach by making them aware of the level of difficulty associated with it and
suggesting strategies that could possibly support them while they are adjusting to this
approach (Khalaf & Zin, 2018). Conclusion Exploring and describing the experiences of the PGDip in TVET participants of the
enquiry-based blended-learning approach used in the programme showed that this
approach to learning was well received by the participants and that they found it well
suited to developing their leadership and managerial skills. The participants acknowledged
that the enquiry-based blended-learning approach influenced their engagement in the
programme and that it enabled them as TVET leaders to function in the South African
context. The challenges that were noted with this approach were mostly related to
contextual factors that could easily be managed and resolved by the facilitators of the
programme. Leaders are not developed overnight and there is not one single approach that
could be regarded as the best to deal positively with the leadership skills crisis in the TVET
sector. However, this study suggests that an enquiry-based blended-learning approach
could be one option that has the potential to develop critical and self-driven leaders in this
sector. Limitations of this study and future research This study was conducted during the COVID-19 lockdown in 2020, when face-to-face
contact learning was primarily converted into online modes. This had implications for the
ways in which the participants evaluated the programme, since they were unable to
experience the entire design of the programme, which would have included in-person face-
to-face learning during week-long contact sessions on campus. They would also have
experienced the scholarly in-person study visit to Germany, which had to be experienced
online instead. To understand fully the ways in which the participants experience the
intended design of the programme, this study may have to be repeated with a subsequent
cohort now that more relaxed lockdown measures permit contact and travel. This may
result in their reporting different experiences, especially their mentoring and networking
experiences. Another limitation that was identified relates to the method of data collection. As stated
previously, the data were collected by two of the facilitators in the programme. In
addition, the participants were asked to report on a programme that the ETDP SETA
and the DHET had funded fully. It is plausible that the participants’ feedback was
mainly positive due to the unequal dynamics that may have existed between participants
and facilitators, and the participants’ need to please the facilitators and their funders
could have been an inhibiting factor. Future research could include a follow-up study
with the participants to gain their perspective on the ways in which this programme
changed and benefitted their leadership skills after applying their newly acquired
knowledge gained from the programme. As noted, the purpose of this study was to explore the subjective experiences of the first
cohort of participants who enrolled for the PGDip programme. The programme design
and the way it influenced their engagement in the programme were considered. From a
programme improvement perspective, this study provided a number of valuable insights. However, the study was limited to experiences that are subjective and contextually bound. Considering the need to advance research relating to the ways in which leadership skills
can be developed in the TVET sector, it would be prudent to conduct studies that
objectively assess the effectiveness of enquiry-based learning and blended learning,
specifically in TVET leadership development programmes. — 152 — TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester Acknowledgements This study was funded by the UP Scholarship of Teaching and Learning (SoTL) grant The authors wish to acknowledge other colleagues in the programme, namely Prof. Salome
Human-Vogel, Dr Celeste Combrinck and Dr Marius Pienaar, for their contributions during
the SoTL grant application and data-collection process. Significant contributions to the development and implementation of the PGDip in TVET
programme were made by the College Lecturer Education Project, part of the TLDCIP, a
partnership programme between the DHET and the EU. Acknowledgement is given to
ETDP SETA and GIZ for funding the programme. The partnership with TUM is recognised,
especially concerning the development of the programme. The authors wish to acknowledge and express their gratitude to the PGDip in TVET
programme participants for their willingness to participate in the study. REFERENCES Alfieri, L, Brooks, PJ, Aldrich, NJ & Tenenbaum, HR. 2011. Does discovery-based instruction
enhance learning? Journal of Educational Psychology, 103(1):1–18. Badenhorst, JW & Radile, RS. 2018. Poor performance at TVET colleges: Conceptualising a
distributed instructional leadership approach as a solution. Africa Education Review,
15(3):91–112. Balkrishen, P & Mestry, R. 2016. The leadership role of campus managers to improve student
achievement in colleges. South African Journal of Higher Education, 30(5):28–47. Balkrishen, P & Mestry, R. 2016. The leadership role of campus managers to improve student
achievement in colleges. South African Journal of Higher Education, 30(5):28–47. Balkrishen, P & Mestry, R. 2016. The leadership role of campus managers to improve student
achievement in colleges. South African Journal of Higher Education, 30(5):28–47. — 153 — Journal of Vocational, Adult and Continuing Education and Training 5(1) 10.14426/jovacet.v5i1.250 Braun, V & Clarke, V. 2020. One size fits all? What counts as quality practice in (reflexive)
thematic analysis? Qualitative Research in Psychology, 18(3):328–352. Choudaha, R & Van Rest, E. 2018. Envisioning pathways to 2030: Megatrends shaping the
future of global higher education and international student mobility. Study Portals. Available
at: https://files.eric.ed.gov/fulltext/ED592843.pdf. [Accessed 20 March 2020]. Chu, SKW, Reynolds, BR, Tavares, NJ, Notari, M & Lee, CQY. 2017. 21st century skills
development through inquiry-based learning. Singapore: Springer. Creswell, JW. 2014. Research design: Qualitative, quantitative, and mixed method approaches (4
ed.). London: Sage. Cronje, J. 2020. Towards a new definition of blended learning. Electronic Journal of
e-Learning, 18(2):114–121. Dewey, J. 1916. Democracy and education. New York: MacMillan. DHET (Department of Higher Education and Training). 2018. Implementation Evaluation of
the Technical and Vocational Education and Training College Expansion and Capacity
Development Programme. [Online]. Available at: https://www.dhet.gov.za/SiteAssets/
PolicyResearchEvaluation/Final%20report%20TVET%20CECDP-6%2004%202018.pdf. [Accessed 11 April 2022]. Furtak, EM, Seidel, T, Iverson, H & Briggs, DC. 2012. Experimental and quasi-experimental
studies of inquiry-based science teaching: A meta-analysis. Review of Educational
Research, 82(3):300–329. Gleason, NW (Ed.). 2018. Higher education in the era of the fourth industrial revolution. Singapore:
Palgrave Macmillan. Gutierez, SB. 2015. Collaborative professional learning through lesson study: Identifying the
challenges of inquiry-based teaching. Issues in Educational Research, 25(2):118–134. HRDC (Human Resource Development Council). 2013. Profile of the TVET sector paper. [Online]. Available at: https://hrdcsa.org.za/2013/08/15/profile-tvet-sector-paper/. [Accessed
11 April 2022]. Jerrim, J, Oliver, M & Sims, S. 2019. The relationship between inquiry-based teaching and
students’ achievement. New evidence from a longitudinal PISA study in England. Learning
and Instruction, 61:35–44. Jonker, H, März, V & Voogt, J. REFERENCES 2018. Teacher educators’ professional identity under construction:
The transition from teaching face-to-face to a blended curriculum. Teaching and Teacher
Education, 71. https://doi.org/10.1016/j.tate.2017.12.016. Khalaf, BK & Zin, ZBM. 2018. Traditional and inquiry-based learning pedagogy: A systematic
critical review. International Journal of Instruction, 11(4):545–564. https://doi.org/10.12973/
iji.2018.11434a. Leijen, Ä, Allas, R, Toom, A, Husu, J, Marcos, JJM, Meijer, P, Knezic, D, Pedaste, M & Krull, E. 2014. Guided reflection for supporting the development of student teachers’ practical
knowledge. Procedia – Social and Behavioral Sciences, 112:314–322. Lewis, P. 2018. Globalizing the liberal arts: Twenty-first-century education in higher education in the
era of the fourth industrial revolution. Singapore: Palgrave Macmillan. Lubua, EW. 2019. Addressing students’ poor performance through blended learning. Information
Technologist, 16(1):137–147. — 154 — TVET leaders’ experiences of an enquiry-based blended-learning programme – T Smit and S Bester Mack, L. 2010. The philosophical underpinnings of educational research. Polyglossia, 19:5–11. Available
at:
https://secure.apu.ac.jp/rcaps/uploads/fckeditor/publications/polyglossia/
Polyglossia_V19_LindsLi.pdf. [Accessed 13 April 2020]. Mohlokoane, M & Coetzer, I. 2007. Towards a leadership model for the effective management of
Further Education and Training colleges. Africa Education Review, 4(1):15–27. Pedaste, M, Mäeots, M, Siiman, LA, De Jong, T, Van Riesen, SA, Kamp, ET, Manoli, CC,
Zacharia, ZC & Tsourlidaki, E. 2015. Phases of inquiry-based learning: Definitions and the
inquiry cycle. Educational Research Review, 14:47–61. Robertson, C & Frick, L. 2018. Conflicting priorities: The dichotomous roles of leadership and
management at TVET colleges. Journal of Vocational, Adult and Continuing Education and
Training, 1(1):71. Thomas, RM. 2003. Blending qualitative and quantitative: Research methods in theses and
dissertations. California: Sage. Tuli, F. 2010. The basis of distinction between qualitative and quantitative research in social
science: Reflection on ontological, epistemological and methodological perspectives. Ethiopian Journal of Education and Science, 6(1):97–108. WEF (World Economic Forum). 2020. The world of jobs report. [Online]. Available at: https://
www3.weforum.org/docs/WEF_Future_of_Jobs_2020.pdf. [Accessed 11 April 2022]. Willis, JW. 2007. Foundations of qualitative research: Interpretive and critical approaches. London:
Sage. Worku, Y. 2019. Leadership and governance factors that affect the quality of education in further
education and training colleges in Gauteng province. African Journal of Democracy and
Governance, 6(2–3):226–246. Zhao, Y & Watterston, J. 2021. The changes we need: Education post COVID-19. Journal of
Educational Change, 22(1):3–12. — 155 —
|
https://openalex.org/W295368082
|
https://europepmc.org/articles/pmc3684864?pdf=render
|
English
| null |
Poly[tri-μ-aqua-diaqua-μ-phosphonoformato-cobalt(II)sodium]
|
Acta crystallographica. Section E
| 2,013
|
cc-by
| 3,786
|
Table 1 Hydrogen bond geometry (A, ). D—H A
D—H
H A
D A
D—H A
O10—H10B O4i
0.82 (1)
1.99 (1)
2.805 (3)
173 (4)
O10—H10A O5ii
0.82 (1)
1.99 (3)
2.736 (3)
151 (5)
O9—H9B O1iii
0.82 (1)
1.87 (1)
2.683 (3)
178 (4)
O9—H9A O5iv
0.82 (1)
1.98 (1)
2.790 (4)
171 (4)
O8—H8B O4v
0.82 (1)
2.24 (2)
2.988 (4)
152 (3)
O8—H8A O2iii
0.82 (1)
1.94 (1)
2.751 (3)
168 (4)
O7—H7B O10vi
0.82 (1)
1.88 (1)
2.703 (4)
177 (4)
O7—H7A O5ii
0.82 (1)
1.96 (2)
2.757 (3)
166 (5)
O6—H6B O8iii
0.82 (1)
2.02 (2)
2.806 (3)
161 (4)
O6—H6A O3ii
0.82 (1)
1.97 (2)
2.698 (3)
148 (4)
Symmetry
codes:
(i)
x þ 1; y þ 3
2; z þ 1
2;
(ii)
x; y þ 1
2; z þ 3
2;
(iii)
x þ 1; y þ 1; z þ 2; (iv) x; y þ 1; z þ 2; (v) x þ 1; y; z; (vi) x; y þ 3
2; z 1
2. Correspondence e-mail: zxq0537@126.com Received 26 March 2013; accepted 9 May 2013 Key indicators: single-crystal X-ray study; T = 223 K; mean (O–C) = 0.004 A˚;
R factor = 0.035; wR factor = 0.066; data-to-parameter ratio = 10.1. The title complex, [CoNa(CO5P)(H2O)5]n, was obtained by
reacting sodium phosphonoformate with cobalt nitrate. The
complex contains cobalt(II) and sodium ions, which are
bridged by the O atoms of two aqua ligands. The CoII ion is
octahedrally coordinated by three phosphonoformato ligands
(one bi- and the other monodentate) and by two O atoms from
the bridging aqua ligands. The sodium cation is hexa-
coordinated by six O atoms from four bridging and two
terminal aqua ligands. The complex molecules are linked to
give a three-dimensional structure by phosphonoformate
ligands bridging CoII atoms and water molecules establishing
cobalt–sodium bridges. O—H O hydrogen bonding between
the aqua ligands and all O atoms of the phosphonoformato
ligand and neighbouring aqua ligands help to consolidate the
packing. Data collection: CrystalClear (Rigaku, 1999); cell refinement:
CrystalClear; data reduction: CrystalStructure (Rigaku/MSC &
Rigaku, 2000); program(s) used to solve structure: SHELXS97
(Sheldrick, 2008); program(s) used to refine structure: SHELXL97
(Sheldrick, 2008); molecular graphics: SHELXTL (Sheldrick, 2008);
software used to prepare material for publication: publCIF (Westrip,
2010). inorganic compounds Data collection
Rigaku Saturn diffractometer
Absorption correction: multi-scan
(REQAB; Jacobson, 1998)
Tmin = 0.636, Tmax = 0.899
3715 measured reflections
1691 independent reflections
1539 reflections with I > 2(I)
Rint = 0.028
Refinement
R[F 2 > 2(F 2)] = 0.035
wR(F 2) = 0.066
S = 1.09
1691 reflections
168 parameters
10 restraints
All H-atom parameters refined
max = 0.37 e A˚ 3
min = 0.45 e A˚ 3 Acta Crystallographica Section E
Structure Reports
Online
ISSN 1600-5368 Acta Crystallographica Section E
Structure Reports
Online
ISSN 1600-5368 Data collection
Rigaku Saturn diffractometer
Absorption correction: multi-scan
(REQAB; Jacobson, 1998)
Tmin = 0.636, Tmax = 0.899 Acta Crystallographica Section E
Structure Reports Related literature Supplementary data and figures for this paper are available from the
IUCr electronic archives (Reference: IM2427). For biological applications of organophosphorus complexes,
see: Xue et al. (2010); Torres Martin de Rosales et al. (2009);
Galanski et al. (2003); Margiotta et al. (2007); Mesri et al. (1996). Table 1 The authors thank the Foundation of State Key Laboratory
Cultivation Base for the Chemistry and Molecular Engi-
neering of Medicinal Resources (CMEMR2012-A11) and the
Research Projects of Guangxi Department of Education
(201010LX200) for financial support. Poly[tri-l-aqua-diaqua-l-phosphono-
formato-cobalt(II)sodium] Xu-Jian Luoa and Xiu-Qing Zhangb* References Galanski, M., Slaby, S., Jakupec, M. A. & Keppler, B. K. (2003). J. Med. Chem. 46, 4946–4951. Jacobson, R. (1998). REQAB. Private communication to the Rigaku
Corporation, Tokyo, Japan. Supplementary data and figures for this paper are available from the
IUCr electronic archives (Reference: IM2427). Xu-Jian Luoa and Xiu-Qing Zhangb* aCollege of Chemistry and Chemical Engineering, Central South University,
Changsha 410083, People’s Republic of China, and bCollege of Chemistry and
Bioengineering, Guilin University of Technology, Guilin 541004, People’s Republic
of China aCollege of Chemistry and Chemical Engineering, Central South University,
Changsha 410083, People’s Republic of China, and bCollege of Chemistry and
Bioengineering, Guilin University of Technology, Guilin 541004, People’s Republic
of China Refinement H atoms of the water molecules were located in a difference Fourier map and refined, with O—H distances restrained to
0.82 Å. Comment Organophosphates have been widely used in medicinal chemistry and life science. They play an important role in life
processes of substance transportation and energy transformation, and are also important for biological substances, such as
ATP, DNA, RNA, etc. Bisphosphonates (BPs) are metabolically stable analogues of pyrophosphates. They have a very
high affinity to calcium ions and therefore show a very strong inhibitory effect on osteoclastic resorption. They are used
as therapeutic agents for several bone-related diseases. Foscarnet and phosphonoacetic acid are known to inhibit viral
DNA polymerase, inhibit the replication of herpes viruses, and also inhibit retroviruses (Mesri et al., 1996). Recently,
several bifunctional metal-phosphonate complexes have been explored (Galanski et al., 2003; Margiotta et al., 2007; Xue
et al., 2010; Torres Martin de Rosales et al., 2009). The molecular structure of the title compound is shown in Fig. 1. Each Co(II) ion is in an octahedral environment
coordinated by two O atoms (O1, O4) from a chelating phosphonoformate ligand, two O atoms (O6, O7) from two
bridging water molecules and two O atoms (O2A, O3B) from two other phosphonoformates. Similarly, the Na(I) ion is
coordinated by four O atoms (O6, O7, O8, O9) from four bridging water molecules and two O atoms (O9C, O10) from
two terminal water ligands. The complex is linked to 3-D structure by phosphonoformate ligands bridging cobalt atoms
and water molecules establishing cobalt sodium bridges (Fig. 2). Experimental An aqueous solution (15 ml) of Co(NO3)2 × 6 H2O (0.145 g, 0.5 mmol) was added dropwisely to an aqueous solution (15
ml) of sodium phosphonoformate (0.180 g, 0.6 mmol) at 323 K. The resulting mixture was refluxed for 3 h, and then the
aqueous solution was allowed to cool down to room temperature. Pink block shaped crystals suitable for X-ray single
diffraction analysis were harvested by slow evaporation (yield, 65%). Experimental Experimental
Crystal data
[CoNa(CO5P)(H2O)5]
Mr = 294.98
Monoclinic, P21=c
a = 8.299 (2) A˚
b = 11.785 (3) A˚
c = 9.769 (3) A˚
= 106.204 (4)
V = 917.5 (4) A˚ 3
Z = 4
Mo K radiation
= 2.13 mm1
T = 223 K
0.30 0.14 0.05 mm Crystal data
[CoNa(CO5P)(H2O)5]
Mr = 294.98
Monoclinic, P21=c
a = 8.299 (2) A˚
b = 11.785 (3) A˚
c = 9.769 (3) A˚
= 106.204 (4) Margiotta, N., Ostuni, R., Teoli, D., Morpurgo, M., Realdon, N., Palazzo, B. &
Natile, G. (2007). Dalton Trans. pp. 3131–3139. g
,
,
,
,
,
,
p
g ,
,
,
,
,
Natile, G. (2007). Dalton Trans. pp. 3131–3139. Mesri, E. A., Cesarman, E., Arvanitakis, L., Rafii, S., Moore, M. A., Posnett, D. V = 917.5 (4) A˚ 3
Z = 4
Mo K radiation
= 2.13 mm1
T = 223 K
0.30 0.14 0.05 mm V = 917.5 (4) A˚ 3
Z = 4
Mo K radiation
= 2.13 mm1
T = 223 K
0.30 0.14 0.05 mm ,
(
)
pp
Mesri, E. A., Cesarman, E., Arvanitakis, L., Rafii, S., Moore, M. A., Posnett, D. N., Knowles, D. M. & Asch, A. S. (1996). J. Exp. Med. 183, 2385–2390. N., Knowles, D. M. & Asch, A. S. (1996). J. Exp. Med. 183, 2385–2390. Rigaku (1999) CrystalClear Rigaku Corporation Tokyo Japan Rigaku (1999). CrystalClear. Rigaku Corporation, Tokyo, Japan. Rigaku/MSC & Rigaku (2000). CrystalStructure. Rigaku/MSC, The Woodands, Rigaku/MSC & Rigaku (2000). CrystalStructure. Rigaku/M Texas, USA, and Rigaku Coporation, Tokyo, Japan. g
p
y
p
Sheldrick, G. M. (2008). Acta Cryst. A64, 112–122. Sheldrick, G. M. (2008). Acta Cryst. A64, 112–122. Torres Martin de Rosales, R., Finucane, C., Mather, S. J. & Blower, P. J. (2009). Chem. Commun. 32, 4847–4849. Westrip, S. P. (2010). J. Appl. Cryst. 43, 920–925. Xue, Z., Lin, M., Zhu, J., Zhang, J., Li, Y. & Guo, Z. (2010). Chem. Commun. 46, 1212–1214. i35 Acta Cryst. (2013). E69, i35 Luo and Zhang doi:10.1107/S1600536813012853 supplementary materials Acta Cryst. (2013). E69, i35 [doi:10.1107/S1600536813012853] Acta Cryst. (2013). E69, i35 [doi:10.1107/S1600536813012853] Computing details Data collection: CrystalClear (Rigaku, 1999); cell refinement: CrystalClear (Rigaku, 1999); data reduction:
CrystalStructure (Rigaku/MSC & Rigaku, 2000); program(s) used to solve structure: SHELXS97 (Sheldrick, 2008);
program(s) used to refine structure: SHELXL97 (Sheldrick, 2008); molecular graphics: SHELXTL (Sheldrick, 2008);
software used to prepare material for publication: publCIF (Westrip, 2010). sup-1 Acta Cryst. (2013). E69, i35 supplementary materials Figure 1
Molecular structure of one repeating unit of the coordination polymer with the atom-numbering scheme displacement
ellipsoids drawn at the 30% probability level. Figure 2 g
Packing diagram of the title compound viewed along c axis. Figure 1 g
Molecular structure of one repeating unit of the coordination polymer with the atom-numbering scheme displacement
ellipsoids drawn at the 30% probability level. Molecular structure of one repeating unit of the coordination polymer with the atom-numbering scheme displacement
ellipsoids drawn at the 30% probability level. sup-2 Acta Cryst. (2013). E69, i35 supplementary materials Figure 2
P
ki
di
f th titl
d i
d l
i Poly[tri-µ-aqua-diaqua-µ-phosphonoformato-cobalt(II)sodium] E69, i35 supplementary materials θmax = 25.5°, θmin = 3.1°
h = −8→10
k = −12→14
l = −11→7
Refinement
Refinement on F2
Least-squares matrix: full
R[F2 > 2σ(F2)] = 0.035
wR(F2) = 0.066
S = 1.09
1691 reflections
168 parameters
10 restraints
Primary atom site location: structure-invariant
direct methods
Secondary atom site location: difference Fourier
map
Hydrogen site location: inferred from
neighbouring sites
All H-atom parameters refined
w = 1/[σ2(Fo2) + (0.0277P)2]
where P = (Fo2 + 2Fc2)/3
(Δ/σ)max = 0.001
Δρmax = 0.37 e Å−3
Δρmin = −0.45 e Å−3 k = −12→14
l = −11→7 Refinement f
Refinement on F2
Least-squares matrix: full
R[F2 > 2σ(F2)] = 0.035
wR(F2) = 0.066
S = 1.09
1691 reflections
168 parameters
10 restraints
Primary atom site location: structure-invariant
direct methods
Secondary atom site location: difference Fourier
map
Hydrogen site location: inferred from
neighbouring sites
All H-atom parameters refined
w = 1/[σ2(Fo2) + (0.0277P)2]
where P = (Fo2 + 2Fc2)/3
(Δ/σ)max = 0.001
Δρmax = 0.37 e Å−3
Δρmin = −0.45 e Å−3 Secondary atom site location: difference Fourier
map
Hydrogen site location: inferred from
neighbouring sites
All H-atom parameters refined
w = 1/[σ2(Fo2) + (0.0277P)2]
where P = (Fo2 + 2Fc2)/3
(Δ/σ)max = 0.001
Δρmax = 0.37 e Å−3
Δρmin = −0.45 e Å−3 Special details Geometry. All e.s.d.'s (except the e.s.d. in the dihedral angle between two l.s. planes) are estimated using the full
covariance matrix. The cell e.s.d.'s are taken into account individually in the estimation of e.s.d.'s in distances, angles and
torsion angles; correlations between e.s.d.'s in cell parameters are only used when they are defined by crystal symmetry. An approximate (isotropic) treatment of cell e.s.d.'s is used for estimating e.s.d.'s involving l.s. planes. Refinement. Refinement of F2 against ALL reflections. The weighted R-factor wR and goodness of fit S are based on F2,
conventional R-factors R are based on F, with F set to zero for negative F2. The threshold expression of F2 > σ(F2) is used
only for calculating R-factors(gt) etc. and is not relevant to the choice of reflections for refinement. R-factors based on F2
are statistically about twice as large as those based on F, and R- factors based on ALL data will be even larger. Poly[tri-µ-aqua-diaqua-µ-phosphonoformato-cobalt(II)sodium] Crystal data
[CoNa(CO5P)(H2O)5]
Mr = 294.98
Monoclinic, P21/c
Hall symbol: -P 2ybc
a = 8.299 (2) Å
b = 11.785 (3) Å
c = 9.769 (3) Å
β = 106.204 (4)°
V = 917.5 (4) Å3
Z = 4
F(000) = 596
Dx = 2.135 Mg m−3
Mo Kα radiation, λ = 0.71075 Å
Cell parameters from 3028 reflections
θ = 3.1–27.5°
µ = 2.13 mm−1
T = 223 K
Block, pink
0.30 × 0.14 × 0.05 mm
Data collection
Rigaku Saturn
diffractometer
Radiation source: fine-focus sealed tube
Graphite monochromator
Detector resolution: 14.63 pixels mm-1
ω scans
Absorption correction: multi-scan
(REQAB; Jacobson, 1998)
Tmin = 0.636, Tmax = 0.899
3715 measured reflections
1691 independent reflections
1539 reflections with I > 2σ(I)
Rint = 0.028 F(000) = 596
Dx = 2.135 Mg m−3
Mo Kα radiation, λ = 0.71075 Å
Cell parameters from 3028 reflections
θ = 3.1–27.5°
µ = 2.13 mm−1
T = 223 K
Block, pink
0.30 × 0.14 × 0.05 mm Absorption correction: multi-scan
(REQAB; Jacobson, 1998)
Tmin = 0.636, Tmax = 0.899
3715 measured reflections
1691 independent reflections
1539 reflections with I > 2σ(I)
Rint = 0.028 sup-3 Acta Cryst. (2013). Special details Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Å2)
x
y
z
Uiso*/Ueq
Co1
0.08112 (5)
0.64526 (3)
0.69444 (4)
0.01133 (13)
P1
−0.00606 (10)
0.38705 (6)
0.65570 (8)
0.01079 (19)
Na1
0.49306 (16)
0.63218 (10)
0.95685 (13)
0.0201 (3)
O1
0.1359 (3)
0.47211 (16)
0.6737 (2)
0.0137 (5)
O2
0.0139 (3)
0.30646 (17)
0.7807 (2)
0.0146 (5)
O3
−0.0498 (3)
0.32183 (16)
0.5160 (2)
0.0147 (5)
O4
−0.1606 (3)
0.58356 (16)
0.6816 (2)
0.0158 (5)
O5
−0.3256 (3)
0.43235 (17)
0.6558 (2)
0.0189 (5)
O6
0.1718 (3)
0.62170 (19)
0.9232 (2)
0.0179 (5)
H6A
0.139 (5)
0.668 (3)
0.971 (4)
0.047 (14)*
H6B
0.164 (4)
0.5600 (16)
0.960 (3)
0.024 (10)*
O7
0.3298 (3)
0.71602 (19)
0.7377 (3)
0.0171 (5)
H7A
0.313 (6)
0.777 (2)
0.770 (5)
0.088 (19)*
H7B
0.388 (4)
0.706 (3)
0.683 (3)
0.029 (11)*
O8
0.7866 (3)
0.6010 (2)
0.9711 (3)
0.0226 (6)
H8A
0.850 (4)
0.636 (3)
1.038 (3)
0.048 (14)*
H8B
0.816 (4)
0.619 (3)
0.901 (2)
0.027 (11)*
O9
0.5412 (3)
0.5383 (2)
1.1731 (3)
0.0189 (5)
H9A
0.473 (3)
0.540 (3)
1.220 (3)
0.031 (12)*
H9B
0.639 (2)
0.537 (3)
1.221 (4)
0.040 (13)*
O10
0.5291 (3)
0.8103 (2)
1.0633 (3)
0.0219 (6)
H10A
0.484 (7)
0.866 (3)
1.017 (5)
0.12 (2)*
H10B
0.620 (3)
0.839 (3)
1.104 (4)
0.043 (13)*
C1
−0.1872 (4)
0.4775 (2)
0.6624 (3)
0.0133 (7) Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Å2)
x
y
z
Uiso*/Ueq l atomic coordinates and isotropic or equivalent isotropic displacement parameters (Å2) sup-4 Acta Cryst. (2013). Special details E69, i35 supplementary materials pp
y
Atomic displacement parameters (Å2)
U11
U22
U33
U12
U13
U23
Co1
0.0135 (2)
0.0099 (2)
0.0113 (2)
−0.00024 (18)
0.00465 (17)
0.00001 (17)
P1
0.0130 (4)
0.0095 (4)
0.0109 (4)
0.0001 (3)
0.0048 (3)
0.0003 (3)
Na1
0.0223 (7)
0.0168 (6)
0.0199 (7)
0.0021 (6)
0.0037 (6)
−0.0001 (5)
O1
0.0123 (11)
0.0095 (10)
0.0190 (12)
−0.0013 (9)
0.0040 (9)
0.0003 (9)
O2
0.0205 (12)
0.0125 (10)
0.0101 (11)
−0.0016 (9)
0.0030 (9)
0.0013 (9)
O3
0.0230 (13)
0.0117 (10)
0.0101 (11)
−0.0018 (9)
0.0055 (10)
−0.0005 (9)
O4
0.0170 (12)
0.0103 (10)
0.0222 (12)
−0.0006 (9)
0.0090 (10)
−0.0009 (9)
O5
0.0135 (12)
0.0158 (11)
0.0286 (14)
−0.0032 (10)
0.0078 (10)
0.0000 (10)
O6
0.0261 (14)
0.0141 (12)
0.0152 (12)
0.0033 (11)
0.0085 (11)
0.0011 (11)
O7
0.0197 (14)
0.0171 (12)
0.0173 (13)
0.0003 (11)
0.0096 (11)
−0.0026 (10)
O8
0.0244 (14)
0.0275 (13)
0.0157 (14)
−0.0062 (11)
0.0054 (12)
−0.0047 (12)
O9
0.0153 (14)
0.0238 (13)
0.0187 (14)
0.0017 (11)
0.0065 (12)
0.0003 (10)
O10
0.0221 (15)
0.0181 (12)
0.0250 (15)
−0.0031 (11)
0.0059 (12)
−0.0010 (11)
C1
0.0145 (17)
0.0158 (16)
0.0098 (17)
0.0013 (14)
0.0038 (14)
−0.0018 (13)
Geometric parameters (Å, º)
Co1—O3i
2.036 (2)
Na1—H7A
2.64 (4)
Co1—O2ii
2.097 (2)
O2—Co1iv
2.097 (2)
Co1—O4
2.104 (2)
O3—Co1i
2.036 (2)
Co1—O1
2.113 (2)
O4—C1
1.274 (3)
Co1—O7
2.156 (3)
O5—C1
1.251 (4)
Co1—O6
2.168 (2)
O6—H6A
0.819 (10)
P1—O1
1.519 (2)
O6—H6B
0.820 (10)
P1—O2
1.519 (2)
O7—H7A
0.817 (10)
P1—O3
1.520 (2)
O7—H7B
0.820 (10)
P1—C1
1.859 (3)
O8—H8A
0.821 (10)
Na1—O9
2.319 (3)
O8—H8B
0.817 (10)
Na1—O10
2.325 (3)
O9—Na1iii
2.351 (3)
Na1—O9iii
2.351 (3)
O9—H9A
0.822 (10)
Na1—O7
2.404 (3)
O9—H9B
0.818 (10)
Na1—O8
2.429 (3)
O10—H10A
0.822 (10)
Na1—O6
2.596 (3)
O10—H10B
0.818 (10)
Na1—Na1iii
3.221 (2)
O3i—Co1—O2ii
89.89 (8)
O8—Na1—Na1iii
82.90 (8)
O3i—Co1—O4
98.89 (9)
O6—Na1—Na1iii
87.00 (7)
O2ii—Co1—O4
86.39 (8)
O9—Na1—H7A
149.2 (10)
O3i—Co1—O1
93.29 (8)
O10—Na1—H7A
72.6 (5)
O2ii—Co1—O1
169.80 (9)
O9iii—Na1—H7A
102.7 (5)
O4—Co1—O1
83.54 (8)
O7—Na1—H7A
17.9 (5)
O3i—Co1—O7
88.26 (9)
O8—Na1—H7A
120.2 (11)
O2ii—Co1—O7
89.84 (9)
O6—Na1—H7A
65.1 (11)
O4—Co1—O7
171.90 (9)
Na1iii—Na1—H7A
141.2 (8)
O1—Co1—O7
99.94 (9)
P1—O1—Co1
117.89 (13)
O3i—Co1—O6
167.17 (9)
P1—O2—Co1iv
134.36 (12)
O2ii—Co1—O6
91.74 (8)
P1—O3—Co1i
137.88 (13) Atomic displacement parameters (Å2)
U11
U22
U33
U12
U13
U23
Co1
0.0135 (2)
0.0099 (2)
0.0113 (2)
−0.00024 (18)
0.00465 (17)
0.00001 (17)
P1
0.0130 (4)
0.0095 (4)
0.0109 (4)
0.0001 (3)
0.0048 (3)
0.0003 (3)
Na1
0.0223 (7)
0.0168 (6)
0.0199 (7)
0.0021 (6)
0.0037 (6)
−0.0001 (5)
O1
0.0123 (11)
0.0095 (10)
0.0190 (12)
−0.0013 (9)
0.0040 (9)
0.0003 (9)
O2
0.0205 (12)
0.0125 (10)
0.0101 (11)
−0.0016 (9)
0.0030 (9)
0.0013 (9)
O3
0.0230 (13)
0.0117 (10)
0.0101 (11)
−0.0018 (9)
0.0055 (10)
−0.0005 (9)
O4
0.0170 (12)
0.0103 (10)
0.0222 (12)
−0.0006 (9)
0.0090 (10)
−0.0009 (9)
O5
0.0135 (12)
0.0158 (11)
0.0286 (14)
−0.0032 (10)
0.0078 (10)
0.0000 (10)
O6
0.0261 (14)
0.0141 (12)
0.0152 (12)
0.0033 (11)
0.0085 (11)
0.0011 (11)
O7
0.0197 (14)
0.0171 (12)
0.0173 (13)
0.0003 (11)
0.0096 (11)
−0.0026 (10)
O8
0.0244 (14)
0.0275 (13)
0.0157 (14)
−0.0062 (11)
0.0054 (12)
−0.0047 (12)
O9
0.0153 (14)
0.0238 (13)
0.0187 (14)
0.0017 (11)
0.0065 (12)
0.0003 (10)
O10
0.0221 (15)
0.0181 (12)
0.0250 (15)
−0.0031 (11)
0.0059 (12)
−0.0010 (11)
C1
0.0145 (17)
0.0158 (16)
0.0098 (17)
0.0013 (14)
0.0038 (14)
−0.0018 (13) Atomic displacement parameters (Å2) sup-5 Acta Cryst. Acta Cryst. (2013). E69, i35 Special details E69, i35 supplementary materials supplementary materia
93.92 (9)
C1—O4—Co1
118.1 (2)
87.32 (8)
Co1—O6—Na1
99.98 (10)
79.02 (9)
Co1—O6—H6A
116 (3)
114.36 (12)
Na1—O6—H6A
113 (3)
114.88 (13)
Co1—O6—H6B
121 (2)
110.56 (12)
Na1—O6—H6B
101 (2)
103.06 (13)
H6A—O6—H6B
105 (4)
103.76 (14)
Co1—O7—Na1
106.63 (11)
109.21 (12)
Co1—O7—H7A
99 (4)
93.19 (10)
Na1—O7—H7A
98 (3)
92.76 (10)
Co1—O7—H7B
121 (3)
173.87 (11)
Na1—O7—H7B
103 (2)
156.76 (11)
H7A—O7—H7B
125 (4)
89.86 (9)
Na1—O8—H8A
112 (3)
85.24 (9)
Na1—O8—H8B
116 (3)
87.73 (10)
H8A—O8—H8B
105 (4)
96.27 (10)
Na1—O9—Na1iii
87.24 (10)
82.52 (10)
Na1—O9—H9A
122 (3)
114.84 (10)
Na1iii—O9—H9A
109 (3)
90.14 (9)
Na1—O9—H9B
115 (3)
95.69 (9)
Na1iii—O9—H9B
104 (3)
85.74 (9)
H9A—O9—H9B
114 (4)
66.63 (9)
Na1—O10—H10A
119 (4)
167.95 (9)
Na1—O10—H10B
125 (3)
46.79 (7)
H10A—O10—H10B
99 (4)
139.96 (10)
O5—C1—O4
123.0 (3)
45.97 (7)
O5—C1—P1
119.6 (2)
126.93 (8)
O4—C1—P1
117.3 (2)
−112.78 (14)
O7—Na1—O6—Co1
20.00 (8)
117.81 (14)
O8—Na1—O6—Co1
−79.7 (5)
−0.87 (16)
Na1iii—Na1—O6—Co1
−112.70 (8)
−96.88 (14)
O3i—Co1—O7—Na1
−154.99 (11)
11.0 (5)
O2ii—Co1—O7—Na1
115.12 (11)
1.70 (13)
O4—Co1—O7—Na1
52.9 (6)
174.31 (13)
O1—Co1—O7—Na1
−61.95 (11)
95.95 (14)
O6—Co1—O7—Na1
23.32 (10)
−157.24 (16)
O9—Na1—O7—Co1
−19.2 (3)
−25.7 (2)
O10—Na1—O7—Co1
−116.96 (11)
91.3 (2)
O9iii—Na1—O7—Co1
66.71 (11)
−43.0 (2)
O8—Na1—O7—Co1
146.19 (10)
−174.31 (17)
O6—Na1—O7—Co1
−20.70 (9)
72.1 (2)
Na1iii—Na1—O7—Co1
45.96 (16)
89.9 (2)
O10—Na1—O9—Na1iii
−178.55 (11)
179.2 (2)
O9iii—Na1—O9—Na1iii
0.0
−2.5 (2)
O7—Na1—O9—Na1iii
84.4 (2)
−118.4 (6)
O8—Na1—O9—Na1iii
−82.39 (9)
−89.3 (2)
O6—Na1—O9—Na1iii
85.74 (9)
−13.2 (4)
Co1—O4—C1—O5
178.7 (2) pp
y
O4—Co1—O6
93.92 (9)
C1—O4—Co1
118.1 (2)
O1—Co1—O6
87.32 (8)
Co1—O6—Na1
99.98 (10)
O7—Co1—O6
79.02 (9)
Co1—O6—H6A
116 (3)
O1—P1—O2
114.36 (12)
Na1—O6—H6A
113 (3)
O1—P1—O3
114.88 (13)
Co1—O6—H6B
121 (2)
O2—P1—O3
110.56 (12)
Na1—O6—H6B
101 (2)
O1—P1—C1
103.06 (13)
H6A—O6—H6B
105 (4)
O2—P1—C1
103.76 (14)
Co1—O7—Na1
106.63 (11)
O3—P1—C1
109.21 (12)
Co1—O7—H7A
99 (4)
O9—Na1—O10
93.19 (10)
Na1—O7—H7A
98 (3)
O9—Na1—O9iii
92.76 (10)
Co1—O7—H7B
121 (3)
O10—Na1—O9iii
173.87 (11)
Na1—O7—H7B
103 (2)
O9—Na1—O7
156.76 (11)
H7A—O7—H7B
125 (4)
O10—Na1—O7
89.86 (9)
Na1—O8—H8A
112 (3)
O9iii—Na1—O7
85.24 (9)
Na1—O8—H8B
116 (3)
O9—Na1—O8
87.73 (10)
H8A—O8—H8B
105 (4)
O10—Na1—O8
96.27 (10)
Na1—O9—Na1iii
87.24 (10)
O9iii—Na1—O8
82.52 (10)
Na1—O9—H9A
122 (3)
O7—Na1—O8
114.84 (10)
Na1iii—O9—H9A
109 (3)
O9—Na1—O6
90.14 (9)
Na1—O9—H9B
115 (3)
O10—Na1—O6
95.69 (9)
Na1iii—O9—H9B
104 (3)
O9iii—Na1—O6
85.74 (9)
H9A—O9—H9B
114 (4)
O7—Na1—O6
66.63 (9)
Na1—O10—H10A
119 (4)
O8—Na1—O6
167.95 (9)
Na1—O10—H10B
125 (3)
O9—Na1—Na1iii
46.79 (7)
H10A—O10—H10B
99 (4)
O10—Na1—Na1iii
139.96 (10)
O5—C1—O4
123.0 (3)
O9iii—Na1—Na1iii
45.97 (7)
O5—C1—P1
119.6 (2)
O7—Na1—Na1iii
126.93 (8)
O4—C1—P1
117.3 (2)
O2—P1—O1—Co1
−112.78 (14)
O7—Na1—O6—Co1
20.00 (8)
O3—P1—O1—Co1
117.81 (14)
O8—Na1—O6—Co1
−79.7 (5)
C1—P1—O1—Co1
−0.87 (16)
Na1iii—Na1—O6—Co1
−112.70 (8)
O3i—Co1—O1—P1
−96.88 (14)
O3i—Co1—O7—Na1
−154.99 (11)
O2ii—Co1—O1—P1
11.0 (5)
O2ii—Co1—O7—Na1
115.12 (11)
O4—Co1—O1—P1
1.70 (13)
O4—Co1—O7—Na1
52.9 (6)
O7—Co1—O1—P1
174.31 (13)
O1—Co1—O7—Na1
−61.95 (11)
O6—Co1—O1—P1
95.95 (14)
O6—Co1—O7—Na1
23.32 (10)
O1—P1—O2—Co1iv
−157.24 (16)
O9—Na1—O7—Co1
−19.2 (3)
O3—P1—O2—Co1iv
−25.7 (2)
O10—Na1—O7—Co1
−116.96 (11)
C1—P1—O2—Co1iv
91.3 (2)
O9iii—Na1—O7—Co1
66.71 (11)
O1—P1—O3—Co1i
−43.0 (2)
O8—Na1—O7—Co1
146.19 (10)
O2—P1—O3—Co1i
−174.31 (17)
O6—Na1—O7—Co1
−20.70 (9)
C1—P1—O3—Co1i
72.1 (2)
Na1iii—Na1—O7—Co1
45.96 (16)
O3i—Co1—O4—C1
89.9 (2)
O10—Na1—O9—Na1iii
−178.55 (11)
O2ii—Co1—O4—C1
179.2 (2)
O9iii—Na1—O9—Na1iii
0.0
O1—Co1—O4—C1
−2.5 (2)
O7—Na1—O9—Na1iii
84.4 (2)
O7—Co1—O4—C1
−118.4 (6)
O8—Na1—O9—Na1iii
−82.39 (9)
O6—Co1—O4—C1
−89.3 (2)
O6—Na1—O9—Na1iii
85.74 (9)
O3i—Co1—O6—Na1
−13.2 (4)
Co1—O4—C1—O5
178.7 (2) sup-6 Acta Cryst. Special details (2013). Special details (2013). E69, i35 supplementary materials Symmetry codes: (ii) −x, y+1/2, −z+3/2; (iii) −x+1, −y+1, −z+2; (v) x+1, −y+3/2, z+1/2; (vi) −x, −y+1, −z+2; (vii) x+1, y, z; (v y+1/2, −z+3/2; (iii) −x+1, −y+1, −z+2; (v) x+1, −y+3/2, z+1/2; (vi) −x, −y+1, −z+2; (vii) x+1, y, z; (viii) x, −y+3/2, z−1/2. supplementary materials O2ii—Co1—O6—Na1
−110.39 (9)
Co1—O4—C1—P1
2.6 (3)
O4—Co1—O6—Na1
163.11 (8)
O1—P1—C1—O5
−177.4 (2)
O1—Co1—O6—Na1
79.78 (9)
O2—P1—C1—O5
−57.9 (3)
O7—Co1—O6—Na1
−20.89 (9)
O3—P1—C1—O5
60.1 (3)
O9—Na1—O6—Co1
−159.41 (9)
O1—P1—C1—O4
−1.1 (3)
O10—Na1—O6—Co1
107.37 (10)
O2—P1—C1—O4
118.4 (2)
O9iii—Na1—O6—Co1
−66.65 (10)
O3—P1—C1—O4
−123.7 (2)
Symmetry codes: (i) −x, −y+1, −z+1; (ii) −x, y+1/2, −z+3/2; (iii) −x+1, −y+1, −z+2; (iv) −x, y−1/2, −z+3/2. Symmetry codes: (i) −x, −y+1, −z+1; (ii) −x, y+1/2, −z+3/2; (iii) −x+1, −y+1, −z+2; (iv) −x, y−1/2, −z+3/2. Hydrogen-bond geometry (Å, º)
D—H···A
D—H
H···A
D···A
D—H···A
O10—H10B···O4v
0.82 (1)
1.99 (1)
2.805 (3)
173 (4)
O10—H10A···O5ii
0.82 (1)
1.99 (3)
2.736 (3)
151 (5)
O9—H9B···O1iii
0.82 (1)
1.87 (1)
2.683 (3)
178 (4)
O9—H9A···O5vi
0.82 (1)
1.98 (1)
2.790 (4)
171 (4)
O8—H8B···O4vii
0.82 (1)
2.24 (2)
2.988 (4)
152 (3)
O8—H8A···O2iii
0.82 (1)
1.94 (1)
2.751 (3)
168 (4)
O7—H7B···O10viii
0.82 (1)
1.88 (1)
2.703 (4)
177 (4)
O7—H7A···O5ii
0.82 (1)
1.96 (2)
2.757 (3)
166 (5)
O6—H6B···O8iii
0.82 (1)
2.02 (2)
2.806 (3)
161 (4)
O6—H6A···O3ii
0.82 (1)
1.97 (2)
2.698 (3)
148 (4)
Symmetry codes: (ii) −x, y+1/2, −z+3/2; (iii) −x+1, −y+1, −z+2; (v) x+1, −y+3/2, z+1/2; (vi) −x, −y+1, −z+2; (vii) x+1, y, z; (viii) x, −y+3/2, z−1/2. Hydrogen-bond geometry (Å, º) Hydrogen-bond geometry (Å, º) sup-7 Acta Cryst. (2013). E69, i35
|
https://openalex.org/W4282916587
|
https://hal.science/hal-03690066v2/document
|
English
| null |
Coronilla juncea, a native candidate for phytostabilization of potentially toxic elements and restoration of Mediterranean soils
|
Scientific reports
| 2,022
|
cc-by
| 13,899
|
Coronilla juncea, a native candidate for
phytostabilization of potentially toxic elements and
restoration of Mediterranean soils Alma Heckenroth, Pascale Prudent, Hélène Folzer, Jacques Rabier, Criquet
Stéven, Arne Saatkamp, Marie-Dominique Salducci, Laurent Vassalo, Isabelle
Laffont-Schwob To cite this version: Alma Heckenroth, Pascale Prudent, Hélène Folzer, Jacques Rabier, Criquet Stéven, et al.. Coro-
nilla juncea, a native candidate for phytostabilization of potentially toxic elements and restoration
of Mediterranean soils. Scientific Reports, 2022, 12, pp.10033. 10.1038/s41598-022-14139-4. hal-
03690066v2 Coronilla juncea, a native candidate
for phytostabilization of potentially
toxic elements and restoration
of Mediterranean soils
OPEN Alma Heckenroth1,2,3, Pascale Prudent4, Hélène Folzer2,3, Jacques Rabier3,
Stéven Criquet3, Arne Saatkamp3, Marie‑Dominique Salducci3, Laurent Vassalo4 &
Isabelle Laffont‑Schwob1,2* Alma Heckenroth1,2,3, Pascale Prudent4, Hélène Folzer2,3, Jacques Rabier3,
Stéven Criquet3, Arne Saatkamp3, Marie‑Dominique Salducci3, Laurent Vassalo4 &
Isabelle Laffont‑Schwob1,2* Soil contamination pattern due to industrial activities often leads to high concentrations of potentially
toxic elements (PTE) decreasing with depth. This spatial heterogeneity of the soil contamination may
have significant consequences on the soil properties and soil living communities. We evaluated the
effects of both surface and solum soil contamination heterogeneity on Coronilla juncea L. (Fabaceae)
functional traits in field conditions and the phytostabilization potential of this species. Plant and soil
samples were collected on 3 sites along a PTE contamination gradient. The correlations between
PTE concentration in plant and soil samples at 2 depths, physico-chemical properties of soil, plant
biomass and soil microbial activity were tested. Field measurements highlight a decreasing PTE
concentration with soil depth in addition to an important surface heterogeneity of As, Cu, Pb, Sb and
Zn soil concentrations. Root PTE concentrations in C. juncea did not follow soil PTE concentrations. Concentrations of PTE in the root parts were higher than those of the aerial parts. Low PTE
translocation and root symbioses with microorganisms suggest that this native plant species may play
a role as engineer species with positive implications for the phytostabilization of Mediterranean PTE
contaminated soils and their ecological restoration. For the two last centuries, industrial activities have generated an accumulation of potentially toxic elements
(PTE)1 in soils at the vicinity of activity areas2. Soil contamination by inorganic compounds coming from slap
heaps and atmospheric deposits is a dissuasive limit for sustainable land uses in many parts of the world3. Indeed,
PTE lead to ecotoxicological effects on plant and soil organisms and, microbial activities that can degrade soil
functions, constrain vegetation recovery and be considered to constitute a human and environmental health
risk. Phytoremediation techniques have been developed for many ecological contexts4. However, little is known
on how spatial heterogeneity in contaminant distribution affects the efficiency of phytoremediation approaches. If industrial contaminations are often characterized by an important spatial heterogeneity5–7, previous research
works mostly focused on topsoil (ca. 0–25 cm) contamination heterogeneity as a whole. The effects of contamina-
tion heterogeneity through the soil profile have been little investigated and approaches taking into consideration
both spatial axes of contamination heterogeneity (surface and solum) are lacking. HAL Id: hal-03690066 L’archive ouverte pluridisciplinaire HAL, est
destinée au dépôt et à la diffusion de documents
scientifiques de niveau recherche, publiés ou non,
émanant des établissements d’enseignement et de
recherche français ou étrangers, des laboratoires
publics ou privés. HAL is a multi-disciplinary open access
archive for the deposit and dissemination of sci-
entific research documents, whether they are pub-
lished or not. The documents may come from
teaching and research institutions in France or
abroad, or from public or private research centers. www.nature.com/scientificreports www.nature.com/scientificreports 1Aix Marseille Univ, IRD, LPED, Marseille, France. 2Fédération de Recherche ECCOREV N° 3098, Aix Marseille Univ,
CNRS, Aix‑en‑Provence, France. 3Aix Marseille Univ, Avignon Université, CNRS, IRD, IMBE, Marseille, France. 4Aix
Marseille Univ, CNRS, LCE, Marseille, France. *email: isabelle.schwob@univ-amu.fr Scientific Reports | (2022) 12:10033 www.nature.com/scientificreports/ Recent works that studied the effects of contamination variability on composition and structure of plant com-
munities found correlations between plant assemblages and PTE soil concentrations indicating an alteration
of the ecosystem trajectory under pollution pressure5,17–19. Plants can deal with small-scale heterogeneity of
contaminants by avoiding the most contaminated patches of soil20. This ‘rooting plasticity’ may have important
consequences for efficiency of phytostabilization in sites where the contaminations are concentrated in topsoils. qfi
y
p y
p
Plant-soil-microorganism interactions in the rhizosphere are determinant for the ability of plant and associ-
ated microorganism communities to develop, by increasing the resistance of the host plant to biotic and abi-
otic stresses21. However, Giller et al.22 showed that microorganisms are more sensitive to stress caused by PTE
contamination compared to other soil organisms. By affecting structure and functional diversity of microbial
communities, PTE are likely to alter the biological processes of soils, such as litter decomposition, soil organic
matter mineralization, and nutrient recycling23,24. Furthermore, inorganic contaminations affect interactions
between plants and microorganisms in the rhizosphere25. Yet, these interactions are a major determinant of the
plant tolerance mechanisms to contaminants, by influencing nutrient availability and PTE bioavailability and
ecotoxicity26 and leading in some cases to a decrease of stress and toxicity generated by PTE27. Indeed, plant-
associated microorganisms can contribute to a reduction of PTE in aboveground parts of plants by promoting
the sequestration of contaminants in both roots and rhizosphere microorganisms28, immobilization in the soil29,
or by direct root accumulation30,31 in continuous feedback linked to plant community composition and soil
abiotic properties32. Phytostabilization approaches are based on the combined action of plants and their associ-
ated microorganisms for stabilization of inorganic contaminants and the physical protection against erosion
provided by plant cover to reduce the transfer of these contaminants33–36. Thus, spatial heterogeneity of soil
contaminants needs to be considered to avoid any failure in reducing PTE transfer. Mechanisms underlying the
strategy of plant-microorganism to deal with contamination heterogeneity have to be elucidated. Moreover, the
length of summer dryness of the soil in certain biogeographic areas such as the Mediterranean Basin added to
steep slopes leading to soil erosion are local constraints increasing the risk of PTE transfer37. www.nature.com/scientificreports/ Besides, a recent
research work on spatial distribution of PTE from smelters from Hunan Province of China contributed to a better
understanding of PTE distribution in soil layers and demonstrated the effect of Zn, Cu, As on microorganism
communities with the help of metagenomic methods38. Microorganisms are a key performance indicator of
the soil quality and may provide information of soil degradation using methods from microbial biomass and
activities to express soil functionalities and/or metagenomic to express soil biodiversity38,39. Consequently, to
optimize phytostabilization of polluted soils, there is a need to take into consideration the spatial heterogeneity
of the soil contamination and plant-microorganism response to PTE in soil40,41. p
g
p
Phytostabilization approaches constitute a nature-based solution considered as adapted to the context of
protected areas where no redevelopment of brownfields is considered. We selected a former lead smelter named
Escalette located in the Calanques National Park (Marseille, south-east France). A previous study highlighted
an important heterogeneity of the PTE topsoil contamination19 and, has led to a selection of native pseudo-
metallophytes being potential candidates for phytostabilization. Pseudo-metallophytes refer to plant species
having acquired PTE tolerance however being able to grow both in PTE polluted and non-polluted areas. They
differ from metallophytes restricted to metal-rich soils42. Amongst those, the native leguminous Coronilla juncea
L. appeared to tolerate high levels of PTE in soils. This species is known to form root symbioses with N-fixing
bacteria favouring rehabilitation of arid degraded lands and considered as a nurse plant43,44. The current study
focused on the phytostabilization potential of this plant species in the context of elevated PTE contamination
heterogeneity in Escalette soils. g
y
We further assume that the PTE soil concentration should be higher in the topsoil layer, as the industrial con-
tamination of the area was mainly caused by atmospheric deposition of contaminants from the smelter activity,
then by those contained in ashes from the degraded horizontal chimney located upstream of the contaminated
studied area38. We were also expecting deleterious effects on microbial and plant traits depending on PTE soil
concentrations39. We hypothesize deleterious effects may follow a hump-back model with intermediary PTE
contamination being the less deleterious on biotic interactions (sensu Grime45).f Our purpose here was to investigate: (i) the effects of both surface and solum contaminant heterogeneities on
root and microorganism functional traits; (ii) the phytostabilization efficiency under field conditions. www.nature.com/scientificreports/ We there-
fore conducted an analysis of PTE concentrations in plant and soil samples, the characterization of pedological
properties and indirect measurements of soil microbial activity based on basal and substrate induced respirations
(BR vs SIR) and estimation of the number of root nodules, depending on soil depth in a range of field conditions. Coronilla juncea, a native candidate
for phytostabilization of potentially
toxic elements and restoration
of Mediterranean soils
OPEN However, the heterogeneity
of soil characteristics even at a small scale is known to influence the response of plants and their associated
microorganisms in terms of structure and functions, from the processes linked to the root growth and nutrient
resource capture to ecosystem functioning8–11. This soil heterogeneity contributes to the characteristic patchy
distribution of vegetation in dry ecosystems, patches that in turn provide more favorable microenvironments
for plant regeneration12. In industrial and urban contexts, spatial heterogeneity of the soil contamination is
often considered as typical7. This is also the case for smelter activities that generate important PTE atmospheric
emissions and then particular deposits at the topsoil surface of the surrounding environment6,13,14. The depth
distribution of industrial contaminants throughout the soil profile shows often decreasing concentrations of PTE
with depth, caused by atmospheric deposits and particle run-off15,16. These surface and solum heterogeneities of
soil contamination may have significant consequences on soil functions and communities of living organisms. 1Aix Marseille Univ, IRD, LPED, Marseille, France. 2Fédération de Recherche ECCOREV N° 3098, Aix Marseille Univ,
CNRS, Aix‑en‑Provence, France. 3Aix Marseille Univ, Avignon Université, CNRS, IRD, IMBE, Marseille, France. 4Aix
Marseille Univ, CNRS, LCE, Marseille, France. *email: isabelle.schwob@univ-amu.fr | https://doi.org/10.1038/s41598-022-14139-4 Scientific Reports | (2022) 12:10033 Scientific Reports | (2022) 12:10033 www.nature.com/scientificreports/ Results Soil physico‑chemical characteristics. All soil samples showed similar pedological properties. Since no
significant effect of soil depth was observed on the pedological properties, only the properties measured in the
topsoil, between 0 and 10 cm depth, are given in Table 1. Cation exchange capacity (CEC) and MgO were sig-
nificantly lower in Escalette 2 (E2) compared to Escalette 1 (E1) and Sormiou (S), respectively. Cation exchange
capacity was positively correlated with total organic carbon (TOC; r = 0.8; p-value ≤ 0.001; Fig. 1). Apart the
higher exchangeable Mg content, no significant difference of concentrations for nutritional elements and organic
matter could be detected for soil from the control site to contaminated soils E1 and E2. The soil texture of all
sites was mostly loam and silty loam (Table 1). Total organic carbon content ranged from 2.8 to 4.8%; however,
no significant differences between sites could be revealed. Considering the results obtained for each site and at
both soil depths (Fig. 1), TOC, total Kjeldahl nitrogen (NTK), cation exchange capacity and exchangeable P
and Ca were altogether positively correlated (p-value ≤ 0.05; Fig. 1). The substrate-induced respiration/soil basal
respiration (SIR/BR) ratio showed the lowest value at the most contaminated site E2, with an average increasing
ratio around 490% vs 790% and 760% for S and E1, respectively (Table 1). Considering all sites and soil depths https://doi.org/10.1038/s41598-022-14139-4 Scientific Reports | (2022) 12:10033 | www.nature.com/scientificreports/ Table 1. Soil pedological properties (mean ± standard deviation) of the studied sites measured between 0 and
10 cm depth and FAO soil classification. Means (n = 5) followed by different letters are significantly different
(Wilcoxon test. p ≤ 0.05). Sites: S = reference site at Sormiou. E1 = Escalette 1 and E2 = Escalette 2. Parameters:
CEC = cation exchange capacity; NTK = total Kjeldahl nitrogen; TC = total carbon; IC = inorganic carbon;
TOC = total organic carbon; SIR/BR = substrate-induced respiration/soil basal respiration. Results Soil pedological properties
Sites
S
E1
E2
pHwater
8.08 ± 0.03a
7.94 ± 0.10a
8.11 ± 0.12a
CEC (meq/100 g)
20.30 ± 3.00ab
24.82 ± 7.74a
16.20 ± 3.52b
Clay (%)
20.92 ± 6.26a
20.00 ± 6.02a
15.76 ± 5.52a
Fine silt (%)
19.08 ± 1.05a
16.3 ± 6.45a
19.36 ± 8.27a
Coarse silt (%)
27.16 ± 6.41a
35.78 ± 5.41a
33.06 ± 7.49a
Fine sand (%)
14.4 ± 3.89a
10.56 ± 3.12a
12.5 ± 2.04a
Coarse sand (%)
18.5 ± 3.79a
17.36 ± 5.75a
19.34 ± 6.06a
CaO (g/kg)
10.86 ± 1.48a
11.09 ± 1.62a
9.03 ± 1.31a
K2O (g/kg)
0.22 ± 0.05a
0.21 ± 0.055a
0.19 ± 0.05a
MgO (g/kg)
0.42 ± 0.10a
0.21 ± 0.06b
0.18 ± 0.06b
Na2O (mg/kg)
36.2 ± 14.52a
28.8 ± 10.38a
36.4 ± 25.70a
P2O5 (mg/kg)
26.6 ± 11.35a
59.8 ± 38.54a
37.8 ± 9.98a
NTK (mg/g)
2.85 ± 0.77a
3.88 ± 2.13a
2.69 ± 0.94a
TC (%)
7.12 ± 2.47a
9.28 ± 2.27a
6.85 ± 1.27a
IC (%)
3.23 ± 1.12a
4.51 ± 1.81a
4.04 ± 0.68a
TOC (%)
3.89 ± 1.48a
4.78 ± 2.66a
2.82 ± 1.04a
SIR/BR (%)
794 ± 51a
757 ± 121a
493 ± 136b
FAO soil classification
Calcisol
Calcisol
Calcisol Table 1. Soil pedological properties (mean ± standard deviation) of the studied sites measured between 0 and
10 cm depth and FAO soil classification. Means (n = 5) followed by different letters are significantly different
(Wilcoxon test. p ≤ 0.05). Sites: S = reference site at Sormiou. E1 = Escalette 1 and E2 = Escalette 2. Parameters:
CEC = cation exchange capacity; NTK = total Kjeldahl nitrogen; TC = total carbon; IC = inorganic carbon;
TOC = total organic carbon; SIR/BR = substrate-induced respiration/soil basal respiration. Table 1. Soil pedological properties (mean ± standard deviation) of the studied sites measured between 0 and
10 cm depth and FAO soil classification. Means (n = 5) followed by different letters are significantly different
(Wilcoxon test. p ≤ 0.05). Sites: S = reference site at Sormiou. E1 = Escalette 1 and E2 = Escalette 2. Parameters:
CEC = cation exchange capacity; NTK = total Kjeldahl nitrogen; TC = total carbon; IC = inorganic carbon;
TOC = total organic carbon; SIR/BR = substrate-induced respiration/soil basal respiration. Table 1. Soil pedological properties (mean ± standard deviation) of the studied sites measured between 0 and
10 cm depth and FAO soil classification. Results Soil PTE
Depth
Sites
S
E1
E2
Al
0–10 cm
31,926 ± 8261a
30,382 ± 10802a
34,621 ± 8900a
10–20 cm
33,149 ± 8591a
27,945 ± 4622a
41,217 ± 10741a
As
0–10 cm
15.5 ± 3.5c
675 ± 262b
5945 ± 1974a
10–20 cm
14.0 ± 1.9b
505 ± 207b
1745 ± 863a
Cu
0–10 cm
15.9 ± 2.3bc
23.7 ± 7.8b
30.5 ± 9.9ab
10–20 cm
14.2 ± 1.5a
20.9 ± 7.5a
19.1 ± 10.3a
Fe
0–10 cm
23,590 ± 4885a
21,904 ± 5133a
23,989 ± 4737a
10–20 cm
19,866 ± 2932a
21,400 ± 5003a
26,337 ± 6222a
Mn
0–10 cm
683 ± 130a
491 ± 252a
634 ± 149a
10–20 cm
713 ± 143a
440 ± 185a
705 ± 156a
Pb
0–10 cm
90.9 ± 36.9c
6286 ± 2811b
42,320 ± 11251a
10–20 cm
59.3 ± 17.9c
5624 ± 3775b
12,159 ± 5932a
Sb
0–10 cm
1.4 ± 1.31c
202 ± 38b
2589 ± 829a
10–20 cm
1.1 ± 0.05c
191 ± 69b
885 ± 359a
Ti
0–10 cm
581 ± 79a
491 ± 88a
522 ± 30a
10–20 cm
558 ± 118a
490 ± 74a
549 ± 67a
Zn
0–10 cm
105 ± 18.2c
1767 ± 760b
10,704 ± 4524a
10–20 cm
96.1 ± 13.5c
1290 ± 720b
3898 ± 833a Table 2. Soil elements (PTE and other element pseudo-total concentrations in mg/kg) measured at 0–10 cm
and 10–20 cm depth. In the same row, values (mean ± standard deviation (n = 5)) followed by different letters
are significantly different (Wilcoxon test. P ≤ 0.05). Sites: S = reference site at Sormiou. E1 = Escalette 1 and
E2 = Escalette 2. Table 2. Soil elements (PTE and other element pseudo-total concentrations in mg/kg) measured at 0–10 cm
and 10–20 cm depth. In the same row, values (mean ± standard deviation (n = 5)) followed by different letters
are significantly different (Wilcoxon test. P ≤ 0.05). Sites: S = reference site at Sormiou. E1 = Escalette 1 and
E2 = Escalette 2. together, there were no significant correlations between BR, SIR neither SIR/BR and, other soil characteristics. But there were significant negative correlations between SIR/BR and As, Pb, Sb and Zn soil pseudo-total concen-
trations (p-value ≤ 0.05; Fig. 1). Soil potentially toxic element contamination heterogeneity. Results Looking to the element concentra-
tions within the topsoil between 0 and 10 cm depth, it was possible to distinguish two groups of elements: on
one hand, As, Cu, Pb, Sb and Zn with concentrations that were significantly different among sites and following
a decreasing trend i.e. E2 > E1 > S, out of which no significant difference of Cu concentrations were pointed out
between E1 and E2 (Table 2); on the other hand, Al, Fe, Mn and Ti soil concentrations with no significant differ-
ence detected between the 3 sites. The analysis of the PTE concentrations globally showed an important hetero-
geneity of topsoils as indicated by high standard deviations (Table 2). As, Cu, Pb, Sb and Zn showed decreasing
concentrations with soil depth (Table 3). Moreover, significant differences were observed for the variation of
PTE concentrations between 0–10 cm and 10–20 cm depth when comparing the most contaminated site E2 and
reference site S (Table 3). Concerning As and Zn, this difference between the two depths was also significant for
all sites. Since this difference of concentration calculated between 0–10 cm and 10–20 cm soil depth quantifies
the PTE contamination heterogeneity through the soil profile, it was quite clear that the most contaminated
sites showed also the most heterogeneous vertical contamination regarding the data (Table 2). More globally,
the contamination heterogeneity, both horizontal and vertical, seemed more marked on the site where PTE
concentrations reached the highest levels. The phytoavailable PTE concentrations showed the same trend as
pseudo-total ones with the highest values observed at the E2 site, then decreasingly E1 and S (only elements that
showed different pseudo-total concentrations from one site to another are presented Table 3). At the reference
site S, phytoavailable concentrations of As and Sb were below the detection limit. For Pb, a phytoavailable frac-
tion of 4% was measured between 0 and 10 cm depth at this site. Cu phytoavailable concentrations were similar
among sites, following the trend of the Cu pseudo-total concentrations. For the other PTE, the phytoavailable
fractions seemed to vary depending on each element, site and depth: at Escalette sites (E1 and E2), phytoavail-
able fractions were between 3 and 9% for Sb, between 9 and 13% for As, 11% and 19% for Cu, between 17 and
21% for Zn, and from 25 to 49% for Pb. Results Means (n = 5) followed by different letters are significantly different
(Wilcoxon test. p ≤ 0.05). Sites: S = reference site at Sormiou. E1 = Escalette 1 and E2 = Escalette 2. Parameters:
CEC = cation exchange capacity; NTK = total Kjeldahl nitrogen; TC = total carbon; IC = inorganic carbon;
TOC = total organic carbon; SIR/BR = substrate-induced respiration/soil basal respiration. Figure 1. Correlogram of Spearman correlation test performed between pedological properties and soil
PTE (metal and metalloid pseudo-total concentrations in mg/kg) measured at both soil depths (0–10 cm and
10–20 cm). Blue dots correspond to the positive correlations and red dots to the negative correlations. The size
and color intensity of the dots are proportional to the correlation coefficient value. Parameters: CEC = cation
exchange capacity; NTK = total Kjeldahl nitrogen; TC = total carbon; IC = inorganic carbon; TOC = total organic
carbon; SIR/BR = substrate-induced respiration/soil basal respiration. Figure 1. Correlogram of Spearman correlation test performed between pedological properties and soil
PTE (metal and metalloid pseudo-total concentrations in mg/kg) measured at both soil depths (0–10 cm and
10–20 cm). Blue dots correspond to the positive correlations and red dots to the negative correlations. The size
and color intensity of the dots are proportional to the correlation coefficient value. Parameters: CEC = cation
exchange capacity; NTK = total Kjeldahl nitrogen; TC = total carbon; IC = inorganic carbon; TOC = total organic
carbon; SIR/BR = substrate-induced respiration/soil basal respiration. Figure 1. Correlogram of Spearman correlation test performed between pedological properties and soil
PTE (metal and metalloid pseudo-total concentrations in mg/kg) measured at both soil depths (0–10 cm and
10–20 cm). Blue dots correspond to the positive correlations and red dots to the negative correlations. The size
and color intensity of the dots are proportional to the correlation coefficient value. Parameters: CEC = cation
exchange capacity; NTK = total Kjeldahl nitrogen; TC = total carbon; IC = inorganic carbon; TOC = total organic
carbon; SIR/BR = substrate-induced respiration/soil basal respiration. https://doi.org/10.1038/s41598-022-14139-4 Scientific Reports | (2022) 12:10033 | www.nature.com/scientificreports/ Table 2. Soil elements (PTE and other element pseudo-total concentrations in mg/kg) measured at 0–10 cm
and 10–20 cm depth. In the same row, values (mean ± standard deviation (n = 5)) followed by different letters
are significantly different (Wilcoxon test. P ≤ 0.05). Sites: S = reference site at Sormiou. E1 = Escalette 1 and
E2 = Escalette 2. Results Variations (δ) of PTE (pseudo-total concentrations) between 0–10 cm and 10–20 cm depth within
the same sample (δ = [PTE]0–10 – [PTE]10–20) and soil phytoavailable PTE concentrations (mean ± standard
deviation (n = 5)). For phytoavailable PTE, the percentage in brackets refers to the ratio of the phytoavailable
PTE concentrations on pseudo-total PTE concentrations ([PTE]phytoavailable/[PTE]pseudo-total). “ < LOD” indicates
values below detection limits. In the same row, values followed by different letters are significantly different
(Wilcoxon test, P ≤ 0.05). Table 4. PTE and other element concentrations measured in the aerial parts (AP) ant root parts (RP) of
the individuals of Coronilla juncea at the different sites (S, E1, E2) and soil depths. The average root part
concentrations (in italics) have been calculated using mean concentrations and root biomasses per depth. For
a same element and a same site, different letters indicate a significant difference between sites, organs and/or
depths (Wilcoxon test, p-value ≤ 0.05). Results Cu phytoavailable fraction ranged from 14 to 19% at the reference site,
and Zn fraction was 11% for both depths at the same site. The analysis of regressions between pseudo-total and
phytoavailable concentrations showed significant positive correlations (p-value ≤ 0.001), such correlations being
higher in the case of Sb and Zn (r2 > 0.9) compared to As and Pb (0.76 < r2 < 0.78). For Cu, the correlation coef-
ficient r2 was around 0.6. Accumulation and transfer of PTE in plants. There were no significant differences between PTE con-
centrations in root parts of C. juncea from different soil depths for both contaminated sites E1 and E2 (Table 4),
in contrast to the PTE concentrations measured in the soils. There were generally higher concentrations of PTE
in the root parts compared to the aerial parts, especially at the most contaminated site E2 and with the major
concentration differences observed between aerial parts and root parts from the topsoil layer (0 to 10 cm). Fur- https://doi.org/10.1038/s41598-022-14139-4 Scientific Reports | (2022) 12:10033 | www.nature.com/scientificreports/ Table 3. Variations (δ) of PTE (pseudo-total concentrations) between 0–10 cm and 10–20 cm depth within
the same sample (δ = [PTE]0–10 – [PTE]10–20) and soil phytoavailable PTE concentrations (mean ± standard
deviation (n = 5)). For phytoavailable PTE, the percentage in brackets refers to the ratio of the phytoavailable
PTE concentrations on pseudo-total PTE concentrations ([PTE]phytoavailable/[PTE]pseudo-total). “ < LOD” indicates
values below detection limits. In the same row, values followed by different letters are significantly different
(Wilcoxon test, P ≤ 0.05). Results Sites
Variations (δ) of PTE soil concentration between 0–10 cm and 10–20 cm
As
Cu
Pb
Sb
Zn
Sormiou (S)
Min
− 0.5
− 0.8
13
− 1.1
− 7.3
Mean ± SD
5.5 ± 5.0 c
1.8 ± 2.5 bc
37 ± 23 bc
1.5 ± 1.7 bc
10.6 ± 20.9 c
Max
11.5
4.8
66
2.6
40.0
Escalette 1 (E1)
Min
86
0.05
− 756
− 50
243
Mean ± SD
151 ± 72 b
3.3 ± 2.6 b
732 ± 1113 b
10.5 ± 44 b
446 ± 223 b
Max
230
6.6
1914
47
643
Escalette 2 (E2)
Min
3670
11.3
25,593
1281
4249
Mean ± SD
4850 ± 1321 a
13.9 ± 4.1 a
34,116 ± 8069 a
1985 ± 614 a
8164 ± 3556 a
Max
6441
20.0
44,991
2714
12,351
Sites
Depths
Phytoavailable PTE (mg kg−1) at 0–10 cm and 10–20 cm soil depth
As
Cu
Pb
Sb
Zn
Sormiou (S)
0–10 cm
< LOD
2.9 ± 1.3 (19%)
5.52 ± 7.01 (4%)
< LOD
11.13 ± 5.37 (11%)
10–20 cm
< LOD
2.0 ± 0.8 (14%)
< LOD
< LOD
10.32 ± 3.4 (11%)
Escalette 1 (E1)
0–10 cm
78.3 ± 43.5 (11%)
4.1 ± 1.3 (17%)
3075 ± 1344 (49%)
7.88 ± 3.57 (4%)
325 ± 184 (17%)
10–20 cm
49.7 ± 35.0 (9%)
3.0 ± 1.1 (14%)
2336 ± 1245 (45%)
4.86 ± 4.18 (3%)
241 ± 144 (19%)
Escalette 2 (E2)
0–10 cm
670 ± 376 (11%)
5.9 ± 2.2 (19%)
10,246 ± 2873 (25%)
234 ± 90 (9%)
2062 ± 918 (20%)
10–20 cm
267 ± 266 (13%)
2.4 ± 2.8 (11%)
4818 ± 3163 (41%)
74 ± 35 (8%)
750 ± 269 (21%) Table 3. Variations (δ) of PTE (pseudo-total concentrations) between 0–10 cm and 10–20 cm depth within
the same sample (δ = [PTE]0–10 – [PTE]10–20) and soil phytoavailable PTE concentrations (mean ± standard
deviation (n = 5)). For phytoavailable PTE, the percentage in brackets refers to the ratio of the phytoavailable
PTE concentrations on pseudo-total PTE concentrations ([PTE]phytoavailable/[PTE]pseudo-total). “ < LOD” indicates
values below detection limits. In the same row, values followed by different letters are significantly different
(Wilcoxon test, P ≤ 0.05). Table 3. Results Site, organ
and depth
PTE and other elements concentrations (mg kg−1)
Al
As
Cu
Fe
Mn
Pb
Sb
Ti
Zn
Sormiou (S)
S AP
326.9 ± 223.1 a
< LOD
9 ± 3.5 a
146.3 ± 65.4 bc
31.4 ± 10.2
a
3.5 ± 4.1 a
< LOD
3.2 ± 1.5 b
70.8 ± 36.6 a
S RP
0–10 cm
345.3 ± 265 a
< LOD
0.8 ± 0.7 b
35 ± 78.3 c
< LOD
1.2 ± 1.2 a
< LOD
4.4 ± 1.9 b
15.3 ± 14.7 b
S RP
10–20 cm
631.8 ± 372.9 a
< LOD
7.2 ± 4.1 a
444.5 ± 269.4 a
17.6 ± 6.4
b
2.4 ± 1.7 a
< LOD
8.8 ± 5.3 a
17.8 ± 15.2 b
Average RP 377.1 ± 331.7
< LOD
2.3 ± 4.4
129.2 ± 277.2
5.0 ± 9.6
1.1 ± 1.4
< LOD
2.8 ± 5.4
14.0 ± 14.1
Escalette 1 (E1)
E1 AP
162.4 ± 79.6 b
4.9 ± 4.1 a
4.2 ± 0.7 a
117.9 ± 51.3 b
26.0 ± 3.7 a 59.7 ± 29.9 a
2.3 ± 1.6 a
2.3 ± 1.3 b
88.6 ± 28.9 a
E1 RP
0–10 cm
739.8 ± 569.4 a
22.8 ± 17.5
a
7.8 ± 4.3 a
482.6 ± 343.1 a
15.8 ± 10.3
a
271.6 ± 183.4
a
6.3 ± 3.7 a
9.9 ± 8.6 a
97.4 ± 62.3 a
E1 RP
10–20 cm
1736.6 ± 32.5 ab
50.5 ± 19.6
a
5.7 ± 1.2 a
1116.9 ± 74.4
ab
22.9 ± 1.9 a 463 ± 251.7 a
13 ± 3.1 a
22.7 ± 1.3
ab
127.1 ± 47.6 a
Average RP 693.6 ± 673.0
22.6 ± 21.2
6.57 ± 3.6
462.0 ± 418.5
13.9 ± 9.1
271.7 ± 204.2
6.1 ± 2.6
8.8 ± 9.4
93.3 ± 56.3
Escalette 2 (E2)
E2 AP
140.1 ± 119.8 b
9.4 ± 11.8 b 3.8 ± 0.9 b
90.9 ± 78.4 b
30.2 ± 7.9 a 86.5 ± 91.1 b
5.4 ± 4.3 b
1.8 ± 1.7 b
138.7 ± 18.7 a
E2 RP
0–10 cm
718.2 ± 341.5 a
96.4 ± 50.4
a
6.6 ± 2.6 a
498.7 ± 241.8 a
16.9 ± 7.3 a 742.4 ± 405.1
a
44.5 ± 23.1
a
10.5 ± 4.7
a
280.7 ± 161.3 a
E2 RP
10–20 cm
1706.6 ± 1105.5
ab
38.3 ± 2.4
ab
11 ± 7.6 ab
1104.3 ± 744.4
ab
30.5 ± 7.8 a 310.2 ± 37.8
ab
35.1 ± 3.4
ab
25.4 ± 14.3
ab
208.7 ± 66.3 a
Average RP 677.3 ± 717.0
71.7 ± 49.9
6.0 ± 4.3
463.2 ± 467.6
16.3 ± 9.5
553.3 ± 392.6
25.8 ± 24.6
10.1 ± 10.0
233.1 ± 138.9 Table 4. Results PTE and other element concentrations measured in the aerial parts (AP) ant root parts (RP) of
the individuals of Coronilla juncea at the different sites (S, E1, E2) and soil depths. The average root part
concentrations (in italics) have been calculated using mean concentrations and root biomasses per depth. For
a same element and a same site, different letters indicate a significant difference between sites, organs and/or
depths (Wilcoxon test, p-value ≤ 0.05). Table 4. PTE and other element concentrations measured in the aerial parts (AP) ant root parts (RP) of
the individuals of Coronilla juncea at the different sites (S, E1, E2) and soil depths. The average root part
concentrations (in italics) have been calculated using mean concentrations and root biomasses per depth. For
a same element and a same site, different letters indicate a significant difference between sites, organs and/or
depths (Wilcoxon test, p-value ≤ 0.05). Table 4. PTE and other element concentrations measured in the aerial parts (AP) ant root parts (RP) of
the individuals of Coronilla juncea at the different sites (S, E1, E2) and soil depths. The average root part
concentrations (in italics) have been calculated using mean concentrations and root biomasses per depth. For
a same element and a same site, different letters indicate a significant difference between sites, organs and/or
depths (Wilcoxon test, p-value ≤ 0.05). https://doi.org/10.1038/s41598-022-14139-4 Scientific Reports | (2022) 12:10033 | www.nature.com/scientificreports/ Table 5. Bioconcentration factors (BCF) i.e. ratios of Coronilla juncea root PTE concentrations vs. soil PTE
pseudo-total concentrations for As, Cu, Pb, Sb and Zn. Min: minimum; Max: maximum; Means (n = 5); SD:
standard deviation. Sites: S = reference site at Sormiou. E1 = Escalette 1 and E2 = Escalette 2. Results Bioconcentration factors (BCF) showed increasing values with increasing soil depth (Table 5),
highlighted by a positive correlation: this relation was significant considering the BCF of As, Pb, Sb and Zn,
for which Spearman’s r was respectively 0.55, 0.55, 0.66 and 0.63 (p-value < 0.05). The BCF were also negatively
correlated to the PTE pseudo-total soil concentrations with r equal to − 0.69, − 0.66, − 0.64, − 0.53 and − 0.62
for As, Cu, Pb, Sb and Zn respectively (p-value < 0.05). There were no significant correlations between the root
concentrations and BCF for the industrial-originated elements As, Pb, Sb and Zn but a positive one for Al, Cu,
Fe, Mn and Ti (0.56 ≤ r ≤ 0.93; p-value < 0.05). Amongst all the BCF, Cu was the element for which the maximum
values were measured. Translocation factors (TF) for the industry-related PTE were the highest for As, Pb, Sb and Zn at the E2
site (Table 6). At the most contaminated site E2, maximum TF values for As, Pb and Sb ranged from 25.60 to
28.95% and seemed lower than maximum TF values of Cu and Zn at 86.76 and 186.05%, respectively. For all
data measured at Escalette sites, root concentrations for As, Cu and Pb were negatively correlated to Zn trans-
location factor (− 0.73 ≤ r ≤ − 0.68; p-value < 0.05). For all correlations calculated between the TF and PTE root
concentrations (within the topsoil), r coefficients were systematically negative but not significant, except for the
ones mentioned above and, also between TF for Cu and Cu and Zn concentrations in the root parts (r = − 0.88
and r = − 0.69 respectively; p-value < 0.05). Effect of the soil contamination heterogeneity on plant development and its associated
microorganisms. Even if the mean values of root biomass seemed higher in all topsoils compared to deeper
soils, it was only significantly different at E2 site (Fig. 2). Considering all sites and depths, positive correlations
were observed between PTE soil concentrations and root biomasses (0.44 ≤ r ≤ 0.58; p-value < 0.05) with no clear
evidence of an inhibition of the root growth by contaminants. Because of the small sample size and an important
variability (i.e. Results Site
S
E1
E2
Depth
0–10 cm
10–20 cm
0–10 cm
10–20 cm
0–10 cm
10–20 cm
BCF As (%)
Min
0.94
5.66
0.69
2.56
Mean ± SD
< LOD
3.23 ± 2.10
7.79 ± 3.02
< LOD
1.69 ± 1.14
4.07 ± 2.14
Max
6.06
9.93
3.59
5.58
BCF Cu (%)
Min
< LOD
18.25
25.68
29.73
11.89
39.04
Mean ± SD
3.77 ± 8.44
34.06 ± 19.25
25.85 ± 0.24
52.16 ± 35.04
23.64 ± 13.31
92.06 ± 74.98
Max
18.87
67.18
26.02
104.04
45.10
145.07
BCF Pb (%)
Min
< LOD
1.71
5.84
0.00
0.74
3.05
Mean ± SD
1.42 ± 1.56
4.16 ± 2.43
8.04 ± 3.11
4.68 ± 3.80
1.76 ± 1.10
4.56 ± 2.15
Max
3.78
7.52
10.23
8.12
3.59
6.08
BCF Sb (%)
Min
0.98
4.99
0.80
4.11
Mean ± SD
< LOD
3.18 ± 1.83
6.82 ± 2.58
< LOD
1.74 ± 1.05
6.46 ± 3.32
Max
5.60
8.64
3.52
8.80
BCF Zn (%)
Min
< LOD
2.44
5.96
8.23
1.39
6.31
Mean ± SD
12.43 ± 13.60
5.25 ± 2.60
7.87 ± 2.71
19.66 ± 18.53
2.75 ± 1.93
9.03 ± 3.85
Max
35.22
8.95
9.79
47.35
6.14
11.76 Table 5. Bioconcentration factors (BCF) i.e. ratios of Coronilla juncea root PTE concentrations vs. soil PTE
pseudo-total concentrations for As, Cu, Pb, Sb and Zn. Min: minimum; Max: maximum; Means (n = 5); SD
standard deviation. Sites: S = reference site at Sormiou. E1 = Escalette 1 and E2 = Escalette 2. Table 5. Bioconcentration factors (BCF) i.e. ratios of Coronilla juncea root PTE concentrations vs. soil PTE
pseudo-total concentrations for As, Cu, Pb, Sb and Zn. Min: minimum; Max: maximum; Means (n = 5); SD:
standard deviation. Sites: S = reference site at Sormiou. E1 = Escalette 1 and E2 = Escalette 2. thermore, the highest plant concentration values of As, Pb, Sb and Zn were always measured in the root parts
located within the topsoil, where the PTE contaminations originated from past industries were also the highest. Nevertheless, no significant differences between PTE mean concentrations of the roots sampled at different
depths were detected. At the reference site S, significant differences were only measured for Cu, Fe, Mn and Ti
between root parts from both depths with higher concentrations in the deepest samples (from 10 to 20 cm below
the surface). Results Sites
S
E1
E2
TF As (%)
Min
5.83
1.61
Mean ± SD
< LOD
67.00 ± 123.45
9.49 ± 9.82
Max
287.75
25.60
TF Cu (%)
Min
0.00
35.58
39.65
Mean ± SD
20.46 ± 68.12
65.59 ± 29.49
61.73 ± 19.71
Max
152.32
98.81
86.76
TF Pb (%)
Min
0.00
11.06
3.58
Mean ± SD
55.75 ± 57.03
56.62 ± 91.14
12.33 ± 10.34
Max
127.06
219.55
28.95
TF Sb (%)
Min
14.80
4.01
Mean ± SD
< LOD
48.19 ± 52.62
13.32 ± 8.66
Max
141.31
26.84
TF Zn (%)
Min
0.00
42.77
22.80
Mean ± SD
402.92 ± 321.34
186.65 ± 198.34
75.76 ± 64.56
Max
685.81
506.08
186.05 Table 6. Translocation factors (TF) in Coronilla juncea individuals i.e. ratios of PTE aerial part concentration
vs. PTE root concentration in the topsoil (0 to 10 cm depth) for As, Cu, Pb, Sb and Zn. Min: minimum; Max:
maximum; Means (n = 5); SD: standard deviation. Sites: S = reference site at Sormiou. E1 = Escalette 1 and
E2 = Escalette 2. Sites
S
E1
E2
TF As (%)
Min
5.83
1.61
Mean ± SD
< LOD
67.00 ± 123.45
9.49 ± 9.82
Max
287.75
25.60
TF Cu (%)
Min
0.00
35.58
39.65
Mean ± SD
20.46 ± 68.12
65.59 ± 29.49
61.73 ± 19.71
Max
152.32
98.81
86.76
TF Pb (%)
Min
0.00
11.06
3.58
Mean ± SD
55.75 ± 57.03
56.62 ± 91.14
12.33 ± 10.34
Max
127.06
219.55
28.95
TF Sb (%)
Min
14.80
4.01
Mean ± SD
< LOD
48.19 ± 52.62
13.32 ± 8.66
Max
141.31
26.84
TF Zn (%)
Min
0.00
42.77
22.80
Mean ± SD
402.92 ± 321.34
186.65 ± 198.34
75.76 ± 64.56
Max
685.81
506.08
186.05 Table 6. Translocation factors (TF) in Coronilla juncea individuals i.e. ratios of PTE aerial part concentration
vs. PTE root concentration in the topsoil (0 to 10 cm depth) for As, Cu, Pb, Sb and Zn. Min: minimum; Max:
maximum; Means (n = 5); SD: standard deviation. Sites: S = reference site at Sormiou. E1 = Escalette 1 and
E2 = Escalette 2. Figure 2. Relative root biomass (mean ± standard deviation) i.e. ratio of the dry root biomass of Coronilla
juncea vs. dry soil mass at different soil depths and sites. Means are expressed in gram of dry roots per kg of dry
soil. Results only one nodule observed in all Sormiou root samples, from 0 to 11 nodules per root samples
in E1 site, and from 0 to 4 nodules, in E2 site), no significant differences were detected on nodules occurrence
between the different conditions. Nevertheless, it was obvious that number of root nodule was more important
in the root samples from Escalette sites than S site: from the 5 individuals of C. juncea at each Escalette site we
collected a total of 40 nodules at E1 site and 15 nodules at E2 vs. 1 nodule at Sormiou site (S). Spearman correla-
tion tests performed from all sites data confirmed these significant relations as nodule occurrence was positively
correlated with pseudo-total soil concentrations of As, Pb, Sb and Zn (r = 0.51; 0.53; 0.46 et 0.53 respectively;
p-value ≤ 0.05). Similar results were obtained for phytoavailable PTE soil concentrations (results not showed;
p-value ≤ 0.05). However, at both Escalette sites, there were negative correlations observed between the number
of nodules and the mobile fraction of As, Cu and Sb (− 0.65 ≤ r ≤ − 0.52; p-value < 0.05), also with pH (r = − 0.53;
p-value < 0.05). Number of root nodules was positively correlated with soil carbon concentration, mostly total
C (r = 0.77; p-value < 0.001) and total organic C (r = 0.76; p-value < 0.001), and NTK (r = 0.73; p-value < 0.001). Furthermore, the distribution of nodules in regard to increasing PTE soil concentrations (expressed as pollution
load index—PLI) seemed to follow a trend (Fig. 3): there were more nodules per root sample at both depths https://doi.org/10.1038/s41598-022-14139-4 Scientific Reports | (2022) 12:10033 | www.nature.com/scientificreports/ Table 6. Translocation factors (TF) in Coronilla juncea individuals i.e. ratios of PTE aerial part concentration
vs. PTE root concentration in the topsoil (0 to 10 cm depth) for As, Cu, Pb, Sb and Zn. Min: minimum; Max:
maximum; Means (n = 5); SD: standard deviation. Sites: S = reference site at Sormiou. E1 = Escalette 1 and
E2 = Escalette 2. Results Different letters indicate a significant difference between means (p-value ≤ 0.05). Sites: S = reference site at
Sormiou. E1 = Escalette 1 and E2 = Escalette 2. Figure 2. Relative root biomass (mean ± standard deviation) i.e. ratio of the dry root biomass of Coronilla
juncea vs. dry soil mass at different soil depths and sites. Means are expressed in gram of dry roots per kg of dry
soil. Different letters indicate a significant difference between means (p-value ≤ 0.05). Sites: S = reference site at
Sormiou. E1 = Escalette 1 and E2 = Escalette 2. at medium level of TPE contaminations (corresponding mostly to Escalette 1 (E1) samples). We observed an
absence of nodules in roots both for most of the non-contaminated samples from Sormiou (S) and for some of
the most contaminated points at Escalette 2 (E2), that could reveal plant tolerance mechanism to face PTE con-
tamination linked to N-fixing bacteria. at medium level of TPE contaminations (corresponding mostly to Escalette 1 (E1) samples). We observed an
absence of nodules in roots both for most of the non-contaminated samples from Sormiou (S) and for some of
the most contaminated points at Escalette 2 (E2), that could reveal plant tolerance mechanism to face PTE con-
tamination linked to N-fixing bacteria. www.nature.com/scientificreports/ w.nature.com/scientificreports/
Figure 3. Number of nodules (black crosses) in root samples at different soil depths in regard to an increasing
pollution load index (red bars) gradient in the soil. On the axis X, the sample code corresponds to the site (first
letter), the individual (first number) and, the depth (second number): i.e. S3.2 = Sormiou. Individual 3. Depth:
10–20 cm. Figure 3. Number of nodules (black crosses) in root samples at different soil depths in regard to an increasing
pollution load index (red bars) gradient in the soil. On the axis X, the sample code corresponds to the site (first
letter), the individual (first number) and, the depth (second number): i.e. S3.2 = Sormiou. Individual 3. Depth:
10–20 cm. Figure 3. Number of nodules (black crosses) in root samples at different soil depths in regard to an increasing
pollution load index (red bars) gradient in the soil. On the axis X, the sample code corresponds to the site (first
letter), the individual (first number) and, the depth (second number): i.e. S3.2 = Sormiou. Individual 3. Depth:
10–20 cm. confirmed that Pb and Zn in soils profiles up to 7 km from the brownfield were directly linked to the industrial
activity whereas Cu contamination of the soils could not be linked to this activity. y
y
Moreover, physico-chemical analyses of the natural geochemical environment conducted at both reference
and contaminated sites and through the solum revealed quite constant values typical to local Mediterranean
alkaline coastal soils48,49. It highlighted a relative similarity concerning the natural composition of the soils at
the scale of the study area, underpinning the choice of the sites for comparing pollution levels. Furthermore,
the properties that appeared to be the most spatially heterogeneous considering both surface soils and solum
were those linked to the industrial contamination, represented by exogenous trace metal and metalloids. Some
parameters linked to physico-chemical and biological properties of the soil showed some differences between
sites, but globally, physico-chemical soil characteristics seemed to have a negligible influence on PTE mobility
and, PTE concentration levels showed little impact on soil physico-chemical characteristics. Those characteristics
were representative of calcareous Mediterranean soils and of results previously obtained in the same biogeo-
graphical context14,50 i.e. www.nature.com/scientificreports/ Mediterranean oligotrophic soils, with low content of nutrients and organic matter, and
high level of carbonates, the latter considered as reducing PTE mobility51 and host a plant cover adapted to this
harsh habitat with majorly stress-tolerant plant species52,53.f j
y
p
p
Our study aimed at identifying deleterious effects of PTE soil concentrations on microbial and C. juncea traits. A significant negative correlation between Sb and respiration (SIR/BR ratio) was measured in agreement with Sb
toxicity effects on local soil microbial communities observed in a previous study in the massif of Marseilleveyre54
corroborating the toxic effect of this PTE recently examined on Chinese soils spiked with Sb at two different
speciations (Sb(III) and Sb(V))55. Moreover, Guillamot et al.54 also highlighted that the Sb toxicity effect on
microbial community was more important on non-contaminated soils involving inherited tolerance mechanisms
for the local soil microbial communities that develop on contaminated sites. It seems consistent with the results
obtained here on SIR/BR: the absence of significant difference between the reference site S and contaminated
site E1 could be related to the tolerance of microbial communities; as SIR/BR appeared to be significantly lower
at E2 site, which could indicate toxicity effects due to higher PTE contamination. 2
yf
g
However, the number of nodules in the C. juncea root systems was not reduced in the most contaminated
site and no significant reduction of C. juncea growth has been revealed depending on PTE. These results may
confirm the PTE tolerance of this pseudo-metallophyte.hi i
p
p y
The tolerance of C. juncea to PTE might be linked to its root symbiosis with nitrogen-fixing bacteria as pre-
viously reported for other leguminous56,57. Our data sustain this idea by the higher number of nodules in root
parts of plants located in the contaminated area. More precisely, we observed an absence of nodules in roots for
most of the samples from the non-contaminated site Sormiou (S) and a small number of nodules for the most
contaminated points at Escalette 2 (E2). The presence of nodules can be correlated to the soil P availability as it
can enhance nodule development in N-fixing legumes58. P was positively correlated to PTE soil concentrations
in this study, these results could also reveal plant and microorganism tolerance mechanisms to PTE contamina-
tion linked to N-fixing bacteria, as well as ecotoxicological effects linked to increasing PTE concentrations59,60. Discussion
fi ld
l Our field results clearly highlighted the spatial distribution of industry-related PTE depending on soil location
and depth. A surface heterogeneity of As, Cu, Pb, Sb and Zn soil concentrations was observed. A decreasing
trend in surface contamination was reported when increasing the distance to the source i.e. the brownfield of
Escalette. Highly contaminated sites also showed a heterogeneous solum PTE contamination with higher levels
of industry-related PTE in topsoils. Such a solum PTE distribution trend was previously demonstrated at a
local scale by Heckenroth et al.19 and clearly correlated to the distance to the ruins of the smelter chimney. Our
results are also in agreement with those mapped at the scale of the massif of Marseilleveyre46 demonstrating
the effect of wind and uneven relief on PTE dispersion. Our results sustain the idea of a highest contamination
of the topsoil layer, which was also observed by Cabrera et al.15 in Spain, Imperato et al.16 in Italy and Li et al.38
in China considering urban and industrial contaminations. Moreover, our results are in agreement with those
also observed in the massif of Marseilleveyre by Gelly et al.47 in which isotopic analyses were conducted and https://doi.org/10.1038/s41598-022-14139-4 Scientific Reports | (2022) 12:10033 | www.nature.com/scientificreports/ www.nature.com/scientificreports/ also assist the reestablishment of the ecosystem61. These traits are characteristic of good candidates for phyto-
stabilization with a plant tolerance to a wide range of soil PTE concentrations. Furthermore, C. juncea may act
as an ecosystem engineer62 by increasing soil nitrogen content via rhizobial nitrogen fixation63. Consequently, it
may function as a nurse species and favour the establishment of other plants species. Padilla et al.64 previously
showed the usefulness of C. juncea for restoration in arid environments. also assist the reestablishment of the ecosystem61. These traits are characteristic of good candidates for phyto-
stabilization with a plant tolerance to a wide range of soil PTE concentrations. Furthermore, C. juncea may act
as an ecosystem engineer62 by increasing soil nitrogen content via rhizobial nitrogen fixation63. Consequently, it
may function as a nurse species and favour the establishment of other plants species. Padilla et al.64 previously
showed the usefulness of C. juncea for restoration in arid environments. Using native plants for in situ phytostabilization purposes implies to get a strong knowledge about plant strate-
gies in the field, which can be assessed by measuring root traits like PTE uptake, biomass and properties linked
to symbiotic microorganisms65. These parameters may vary in relation to spatial heterogeneity contamination,
which can be both of surface and solum. Under field conditions, response of plants to this spatial heterogeneity
will also determinate their suitability and efficiency for PTE stabilization. fi
Many phytoremediation projects focus on response of plants in pot experiments using homogeneous soil
samples under controlled conditions42,66,67. However, they seem little representative of the high heterogeneity
under field conditions in contaminated areas or the complexity of biotic interactions. In the context of the highly
constrained field conditions of our study area, i.e. shallow soil characterized as “skeletal”, high contamination by
PTE and semi-arid climatic conditions37, it is necessary to go further in the understanding of microsite condi-
tions and related responses of the plant and their associated microorganisms at different soil depths to select an
adapted phytostabilization strategy. Furthermore, it is a known response of individual plants to use avoidance
strategies in relation to heterogeneous soil contamination: for instance, in the case of atmospherically deposited
PTE contaminants, topsoil layer where the pollutants are the most concentrated may be avoided by roots20, and
consequently phytostabilization will not be optimal.h q
y
y
The sheer presence of C. Methods
All h All the experiments comply with relevant institutional, national, and international guidelines and legislation. All the experiments comply with relevant institutional, national, and international guidelines an Study sites and sampling. We studied two sites in the Calanques National Park (43.21357° N, 5.36735°
E) at Sormiou and Escalette (Fig. 4), composed by a mosaic of spontaneous plant communities that corre-
spond to calcareous xero-thermophilous shrublands (Mediterranean matorral), grasslands and scarce stands of
Pinus halepensis Mill. Both sites have a semi-arid, Thermo-Mediterranean climate with average annual rainfall
of 500 mm and average annual temperature of 16 °C69,70. Sites are at 120 to 140 m altitude. Escalette site is a
brownfield of a former lead (Pb) smelter. PTE transfers, mostly composed of Pb, zinc (Zn), arsenic (As), copper
(Cu) and antimony (Sb), are generated by erosion of the contaminated soils and degraded smelter buildings,
under the influence of local strong winds and water runoff. Cadmium (Cd) has not been identified as a PTE
in this study due to its soil content below level of detection in all previous studies. Previous studies at Escalette
site and surroundings showed a high heterogeneity of surface soil contamination and high PTE tolerance of the
native vegetation14,19,71–75.i Using previous measurements of PTE topsoil concentrations and phyto-ecological inventories on the field14,
we preselected measure points where C. juncea individuals were abundant. We conducted measurements of
the topsoil PTE contamination at the foot of the plants using a portable X-ray fluorescence analyzer (Niton®,
xl3t GOLDD; Thermo Scientific). In order to avoid extreme values, only the points corresponding to Pb con-
centrations between the first and third quartiles have been conserved. We discarded plants growing directly
in calcareous rocks. Sampling points were split into two groups at the Escalette site (Fig. 4a,c): “Escalette 1”
(E1) corresponding to the lowest PTE concentrations and “Escalette 2” (E2) corresponding to the highest PTE
concentration range located downstream of the degraded horizontal chimney. We used a third site at Sormiou
(S) as the reference (Fig. 4a,b) representative of the local PTE soil background concentrations in the Calanques
National Park. All sampling (soil and plants) were done according to the authorization given par the Calanques
national park which restricts scientific sampling on its protected area.i pi
p
g
p
For each study site, five replicates corresponding to a whole plant-soil sampling were selected. Aerial parts
of C. juncea were cut at the collar and collected. www.nature.com/scientificreports/ juncea on the extremely contaminated Escalette site already demonstrated its toler-
ance to PTE contamination in a previous study on plant communities19. The results of this study highlighted the
ability of C. juncea to tolerate contaminants and to grow in a wide range of soil PTE concentrations. PTE con-
tamination exerted neither significant nor visible negative effects on plant biomass, even under increasing uptake
of PTE by the plant. Bioconcentration factors (BCF) in aerial parts were always < 1 for industrial PTE, revealing
restricted transfer of PTE from soil to roots, depending on soil PTE concentration level. The lowest translocation
factors (TF), i.e. transfer of PTE from roots to shoots, were measured at the most contaminated site E2. Thus,
the translocation restriction seemed to evolve depending on the concentration of PTE contained in the roots.f g
Many studies on contaminated soils only consider the effects of single stressors on isolated endpoints68. Our
study integrated a pool of PTE, some of them originating from industrial activities, and considered two biologi-
cal components i.e. plants and soil microorganisms. C. juncea tolerance to PTE was confirmed, as well as the
soil resilience ability in the context of both Escalette sites, even if some toxic effects seemed to limit microbial
activity at the most contaminated site E2. Conclusions In a context of high surface and solum heterogeneity of soil contaminants from past industrial activities, Coronilla
juncea seems a good candidate for phytostabilization in its Mediterranean distribution area. Low translocation
of contaminants and root symbiosis with nitrogen fixing microorganisms suggest that Coronilla juncea may play
a prime role for the restoration of contaminated ecosystems at Escalette and other Mediterranean brownfields
by promoting soil quality. www.nature.com/scientificreports/ i
g
gf
g
Our results demonstrated that root PTE concentrations in C. juncea did not follow the solum PTE concentra-
tions since no significant differences of PTE accumulation in roots were observed between the different depths
from the same site (E1 or E2) despite a gradient of soil contamination. Moreover, concentrations of PTE in the
root parts were higher than those of the aerial parts. C. juncea, by reducing PTE transfer into the food web, may Scientific Reports | (2022) 12:10033 | https://doi.org/10.1038/s41598-022-14139-4 www.nature.com/scientificreports/ Methods
All h We then sampled roots and soil at a 15 cm radius around the Scientific Reports | (2022) 12:10033 | https://doi.org/10.1038/s41598-022-14139-4 www.nature.com/scientificreports/ Figure 4. Location of the Escalette and Sormiou study areas (a); location of the sampling points S at the
reference site Sormiou (b) and location of the sampling points E1 and E2 at the Escalette contaminated site (c). Calanques NP = Calanques National Park area. Map created using QGIS including data collected by PJ Dumas
and A Heckenroth. Figure 4. Location of the Escalette and Sormiou study areas (a); location of the sampling points S at the
reference site Sormiou (b) and location of the sampling points E1 and E2 at the Escalette contaminated site (c). Calanques NP = Calanques National Park area. Map created using QGIS including data collected by PJ Dumas
and A Heckenroth. collar: two samples of roots and soil, from 0 to 10 cm depth, and from 10 to 20 cm depth. We kept all plant and
soil samples at 4 °C in the lab until analysis. Soil samples were sieved through a 2 mm sieve and root parts were
isolated from the soil with laboratory forceps. collar: two samples of roots and soil, from 0 to 10 cm depth, and from 10 to 20 cm depth. We kept all plant and
soil samples at 4 °C in the lab until analysis. Soil samples were sieved through a 2 mm sieve and root parts were
isolated from the soil with laboratory forceps. Soil analysis. Soil physico‑chemical characteristics. Soil samples were dried at room temperature for the
analysis of physico-chemical properties and PTE analyses. pH (ISO 10390), total Kjeldahl nitrogen (NTK, ISO
11261) and total organic carbon contents (TOC, ISO 10694) were analyzed by Laboratoire Développement Mé-
diterranée (COFRAC accreditation n° 1-5865). Soil texture was determined using five fractions without decar-
bonatation (NF X31-107): clay (< 2 µm), fine silt (2 µm to 20 µm), coarse silt (20 µm to 50 µm), fine sand (50 µm
to 0.2 mm) and coarse sand (0.2 mm to 2 mm). Available phosphorus (ISO 11263), exchangeable cations (CaO,
K2O, MgO, Na2O, ISO 23470) and cation exchange capacity (CEC, ISO 23470) were also determined by Labora-
toire Développement Méditerranée (COFRAC accreditation n° 1-5865). An aliquot of each soil sample was kept
for PTE analyses as pseudo-total and EDTA extractable fraction. Soil potentially toxic element analysis. www.nature.com/scientificreports/ used. Furthermore, to obtain an assessment of phytoavailable PTE, we used a solution of EDTA (0.05 mol L−1)
with pH adjusted to 7 at a ratio of dry soil/EDTA corresponding to 1/10 w/v76. The solution was agitated at room
temperature in an orbital shaker at 125 rpm for 1 h (Fisher Bioblock Scientific SM30B). Then, the EDTA extracts
were centrifuged for 10 min at 8000 rpm (JP Selecta Medifriger BL-S). All the mineralization and extract prod-
ucts were filtered with a 0.45 μm-mesh, and the PTE concentrations were determined by inductively coupled
plasma-atomic emission spectroscopy (ICP-AES, Jobin Yvon Horiba Spectra 2000). Quality assurance–quality
controls and accuracy were checked using standard soil reference materials (CRM049–050, from RTC, USA)
with accuracies within 100 ± 10%. We calculated contamination factors from PTE concentrations measured in E1
and E2 compared to the background PTE. Then, these contamination factors were used to estimate a multi-con-
tamination level expressed as a pollution load index (PLI) allowing to estimate a multi-contamination level34,69,72. Soil microbial activity. We estimated soil microbial activity by measuring the soil basal respiration (BR), i.e. CO2 released, using 10 g of sub-samples at 60% of their water holding capacity (WHC) weighed in 117 mL glass
flasks (presumed saturating quantity). After internal atmosphere replacement, flasks were closed hermetically
and incubated for 60 min at 20 °C. The CO2 released in the flasks was measured by gas chromatography (GC,
Chrompack CHROM 3-CP 9001) and with a PorapakTM column containing 650 mL h−1 helium circulating. The
substrate-induced respiration (SIR), i.e. CO2 released when substrate is added (g CO2 h−1 g−1 DW), was meas-
ured according to Anderson and Domsch77, using same method than for BR and by adding 50 mg of powdered
glucose to each soil sample. Samples were pre-incubated for 90 min to reach a maximum rate of SIR. Then, after
internal atmosphere replacement, flasks were closed hermetically and incubated for 60 min at 20 °C. The CO2
released in the flasks was measured in the same way than for BR. The growth of microbial activity was estimated
by calculating the ratio between basal and substrate-induced respiration SIR/BR. Plant and root nodule analyses. Roots were separated from soil samples, washed under tap water to
remove soil particles, then carefully rinsed with deionized water and dried using paper towel. For each root
sample, the total number of nodules has been reported. Root and aerial parts of C. www.nature.com/scientificreports/ juncea were weighted within
few hours of the return from the field to obtain the fresh biomass. The plant samples were dried at 60 °C for at
least 72 h and weighed again to obtain the dry biomass. To analyze the plant PTE concentration, the dry samples
were ground at 0.5 mm: the aerial parts using tungsten carbide blades (Foss Tecator Cyclotec 1093) and the root
parts using a mixer mill (Retsch MM400). About 0.5 g dry matter of each plant sample (triplicates) was digested
in microwave mineralizer system (Milestone Start D) with an acidic mixture (2/3 HNO3 + 1/3 HCl). After a fil-
tration using a 0.45 µm mesh, the solutions were analyzed by ICP-AES as previously described for soil samples. Standard plant reference material (DC 73349 from NCS, China) was analyzed as a part of the quality assurance-
quality control protocol (accuracies within 100 ± 10%). Data processing and statistical analyzes. In order to estimate the variation of soil PTE pseudo-total
concentrations, we calculated differences of concentration (delta values) between samples of 0–10 cm and
10–20 cm depth within a plot (δ = [PTE]0–10 − [PTE]10–20). For comparing among sites, we chose to use delta
values rather than directly soil pseudo-total concentrations per depth, that showed a high intra-site variability. Root bioconcentration factors (BCF) were calculated from the soil and root PTE pseudo-total concentra-
tions as following67: BCFroot = [PTE]root parts/[PTE]soil where [PTE] is the PTE pseudo-total concentration. The
translocation factor (TF) was estimated as following: TF = [PTE]aerial parts/[PTE]root part.t g
p
p
We tested for normality using a Shapiro–Wilk test in R 3.0.2. software78 and stats and ade4 packages79. When
data were not normally distributed, we used Wilcoxon tests to analyze the differences between samples from
different sites and/or depths (soil and root samples). We also used Spearman rank correlation tests to analyze
the correlations between variables. Received: 21 November 2021; Accepted: 18 May 2022 Received: 21 November 2021; Accepted: 18 May 2022 Methods
All h Soil dried samples were ground by a RETSCH zm 1000 grinder with
tungsten blades and titanium sieve to 0.2 mm particles, before analyses of the PTE concentrations. To obtain
pseudo-total PTE extracts, we mineralized soils in three replicates in a microwave mineralizer (Milestone Start
D) using aqua regia (1/3 HNO3 + 2/3 HCl). A ratio of dry soil/aqua regia solution corresponding to 1/20 w/v was https://doi.org/10.1038/s41598-022-14139-4 Scientific Reports | (2022) 12:10033 | www.nature.com/scientificreports/ Data availability All data generated or analysed during this study are included in the published article available on request. More
information is available from the corresponding author on reasonable request. Collection of plant material was
done in agreement with the PNCal regulation number 02015-046 available at http://www.calanques-parcnation
al.fr/fr/download/file/fid/1095. www.nature.com/scientificreports/ Ectomycorrhizal fungi and associated bacteria provide protection against heav
(Pinus sylvestris L.) seedlings. Water Air Soil Pollut. 182, 83–90 (2007). 28. Krupa, P. & Kozdrój, J. Ectomycorrhizal fungi and associated bacteria provide protection against heavy metals in inoculated pine
(Pinus sylvestris L.) seedlings. Water Air Soil Pollut. 182, 83–90 (2007). 9. Janoušková, M., Pavlíková, D. & Vosátka, M. Potential contribution of arbuscular mycorrhiza to cadmium immobilisation in soil
Chemosphere 65, 1959–1965 (2006).f 0. Leyval, C., Turnau, K. & Haselwandter, K. Effect of heavy metal pollution on mycorrhizal colonization and function: Physiological
ecological and applied aspects. Mycorrhiza 7, 139–153 (1997).h g
pp
p
y
31. Zhang, Y., Zhang, Y., Liu, M., Shi, X. & Zhao, Z. Dark septate endophyte (DSE) fungi isolated from metal polluted soils: Their
taxonomic position, tolerance, and accumulation of heavy metals in vitro. J. Microbiol. 46, 624–632 (2008). 31. Zhang, Y., Zhang, Y., Liu, M., Shi, X. & Zhao, Z. Dark septate endophyte (DSE) fungi isolated from metal polluted soils: Their
t
i
iti
t l
d
l ti
f h
t l i
it
J Mi
bi l 46 624 632 (2008) hang, Y., Zhang, Y., Liu, M., Shi, X. & Zhao, Z. Dark septate endophyte (DSE) fungi isolated from metal polluted soils: Their
axonomic position, tolerance, and accumulation of heavy metals in vitro. J. Microbiol. 46, 624–632 (2008). 2. Krumins, J. A., Goodey, N. M. & Gallagher, F. Plant–soil interactions in metal contaminated soils. Soil Biol. Biochem. 80, 224–231
(2015). 3. Glick, B. R. Phytoremediation: Synergistic use of plants and bacteria to clean up the environment. Biotechnol. Adv. 21, 383–393
(2003).f (
)
34. Heckenroth, A. et al. What are the potential environmental solutions for diffuse pollution ? In Pollution of Marseille’s Industrial
Calanques—The Impact of the Past on the Present 291–328 (REF2C, 2016). h
5. Li, M. S. Ecological restoration of mineland with particular reference to the metalliferous mine wasteland in China: A review o
research and practice. Sci. Total Environ. 357, 38–53 (2006). research and practice. Sci. Total Environ. 357, 38 53 (2006). 36. Mendez, M. O. & Maier, R. M. Phytoremediation of mine tailings in temperate and arid environments. Rev. Environ. Sci. Biotechnol. 7, 47–59 (2008) 6. Mendez, M. O. & Maier, R. M. Phytoremediation of mine tailings in temperate and arid environments. Rev. Environ. Sci. Biotechnol
7, 47–59 (2008).f 37. Yaalon, D. H. www.nature.com/scientificreports/ Soils in the Mediterranean region: What makes them different?. CATENA 28, 157–169 (1997). f
38. Li, S. et al. A comprehensive survey on the horizontal and vertical distribution of heavy metals and microorganisms in soils of a
Pb/Zn smelter. J. Hazard. Mater. 400, 123255 (2020).f 9. Pérez-de-Mora, A. et al. Microbial community structure and function in a soil contaminated by heavy metals: Effects of plan
growth and different amendments. Soil Biol. Biochem. 38, 327–341 (2006).fifi gf
0. Keller, C. et al. Root development and heavy metal phytoextraction efficiency: Comparison of different plant species in the field
Plant Soil. 249, 67–81 (2003). (
)
41. Lambrechts, T. et al. Comparative analysis of Cd and Zn impacts on root distribution and morphology of Lolium perenne and
Trifolium repens: Implications for phytostabilization. Plant Soil 376, 229–244 (2014).f f
p
y
2. Pauwels, M., Frérot, H., Bonnin, I. & Saumitou-Laprade, P. A broad-scale analysis of population differentiation for Zn tolerance
i
i
d l
i
f
t l
t d
A bid
i h ll i (B
i
) J E
l Bi l 19 1838 1850 (2006) 42. Pauwels, M., Frérot, H., Bonnin, I. & Saumitou-Laprade, P. A broad-scale analysis of population differentiation for Zn tolerance
in an emerging model species for tolerance study: Arabidopsis halleri (Brassicaceae). J. Evol. Biol. 19, 1838–1850 (2006).h 42. Pauwels, M., Frérot, H., Bonnin, I. & Saumitou Laprade, P. A broad scale analysis of population differentiation for Zn tolerance
in an emerging model species for tolerance study: Arabidopsis halleri (Brassicaceae). J. Evol. Biol. 19, 1838–1850 (2006). 43 P dill
F M & P
i
F I Th
l
f
l
t i th
t
ti
f d
d d
i
t F
t E
l E
i
4 196 202 in an emerging model species for tolerance study: Arabidopsis halleri (Brassicaceae). J. Evol. Biol. 19, 1838–1850 (2006). 3. Padilla, F. M. & Pugnaire, F. I. The role of nurse plants in the restoration of degraded environments. Front. Ecol. Environ. 4, 196–202
(2006). 44. Robles, A. B., Allegretti, L. I. & Passera, C. B. Coronilla juncea is both a nutritive fodder shrub and useful in the rehabilitation of
abandoned Mediterranean marginal farmland. J. Arid Environ. 50, 381–392 (2002). g
45. Grime, J. P. Plant Strategies and Vegetation Processes (Wiley, 1979).ff 45. Grime, J. P. Plant Strategies and Vegetation Processes (Wiley, 1979).ff 46. Laffont-Schwob, I. et al. References 1. Pourret, O. & Hursthouse, A. It’s time to replace the term “heavy metals” with “potentially toxic elements” when reporting envi-
ronmental research. IJERPH 16, 4446 (2019). 1. Pourret, O. & Hursthouse, A. It’s time to replace the term “heavy metals” with “potentially toxic elements” when reporting envi-
ronmental research. IJERPH 16, 4446 (2019). 2. Wuana, R. A. & Okieimen, F. E. Heavy metals in contaminated soils: A review of sources, chemistry, risks and best available strate-
gies for remediation. ISRN Ecol. 2011, 1–20 (2011). g
3. Mahar, A. et al. Challenges and opportunities in the phytoremediation of heavy metals contaminated soils: A review. Ecotoxicol. Environ. Saf. 126, 111–121 (2016).i g
3. Mahar, A. et al. Challenges and opportunities in the phytoremediation of heavy metals contaminated soils: A review. Ecotoxicol
Environ. Saf. 126, 111–121 (2016).i g
3. Mahar, A. et al. Challenges and opportunities in the phytoremediation of heavy metals contaminated soils: A review. Ecotoxicol. Environ. Saf. 126, 111–121 (2016).i f
4. Vangronsveld, J. et al. Phytoremediation of contaminated soils and groundwater: Lessons from the field. Environ. Sci. Pollut. Res. 16, 765–794 (2009). f
4. Vangronsveld, J. et al. Phytoremediation of contaminated soils and groundwater: Lessons from the field. Environ. Sci. Pollut. Res
16, 765–794 (2009). f
4. Vangronsveld, J. et al. Phytoremediation of contaminated soils and groundwater: Lessons from the field. Environ. Sci. Pollut. Res. 16, 765–794 (2009). 5. Desjardins, D., Nissim, W. G., Pitre, F. E., Naud, A. & Labrecque, M. Distribution patterns of spontaneous vegetation and pollution
at a former decantation basin in southern Québec, Canada. Ecol. Eng. 64, 385–390 (2014). g
6. Marchiol, L. et al. Gentle remediation at the former “Pertusola Sud” zinc smelter: Evaluation of native species for phytoremediation
purposes. Ecol. Eng. 53, 343–353 (2013). g
6. Marchiol, L. et al. Gentle remediation at the former “Pertusola Sud” zinc smelter: Evaluation of native species for phytoremediation
purposes. Ecol. Eng. 53, 343–353 (2013). 6. Marchiol, L. et al. Gentle remediation at the former “Pertusola Sud” zinc smelter: Evaluation of native species for phytoremediation
purposes. Ecol. Eng. 53, 343–353 (2013). Scientific Reports | (2022) 12:10033 | https://doi.org/10.1038/s41598-022-14139-4 www.nature.com/scientificreports/ www.nature.com/scientificreports/ 7. van Oort, F. et al. Les pollutions métalliques d’un site industriel et des sols environnants : distributions hétérogènes des métaux et
relations avec l’usage des sols. In: Contaminations métalliques des agrosystèmes et écosystèmes péri-urbains 15–44 (Editions Quae,
2009). )
ge, A. Plastic plants and patchy soils. J. Exp. Bot. 57, 401–411 (200 8. Hodge, A. Plastic plants and patchy soils. J. Exp. Bot. 57, 401– g
p
p
y
p
(
)
9. Huber-Sannwald, E. & Jackson, R. B. Heterogeneous soil-resource distribution and plant responses—from individual-plant growth
to ecosystem functioning. In Progress in Botany Vol. 62 (eds Esser, K. et al.) 451–476 (Springer, 2001). g
p
p
y
p
9. Huber-Sannwald, E. & Jackson, R. B. Heterogeneous soil-resource distribution and plant responses—from individua
to ecosystem functioning In Progress in Botany Vol 62 (eds Esser K et al ) 451 476 (Springer 2001) 9. Huber-Sannwald, E. & Jackson, R. B. Heterogeneous soil-resource distribution and plant responses—from individual-plant growth
to ecosystem functioning. In Progress in Botany Vol. 62 (eds Esser, K. et al.) 451–476 (Springer, 2001). 0. Loecke, T. D. & Philip Robertson, G. Soil resource heterogeneity in the form of aggregated litter alters maize productivity. Plan
Soil 325, 231–241 (2009). 1. Reynolds, H. L., Hungate, B. A., Iii, F. S. C. & D’Antonio, C. M. Soil Heterogeneity and Plant Competition in an Annual Grassland
16 (2021). (
)
12. Maestre, F. T., Cortina, J., Bautista, S., Bellot, J. & Vallejo, R. Small-scale environmental heterogeneity and spatiotemporal dynamics
of seedling establishment in a semiarid degraded ecosystem. Ecosystems 6, 630–643 (2003).h 3. Shutcha, M. N. et al. Three years of phytostabilisation experiment of bare acidic soil extremely contaminated by copper smelting
using plant biodiversity of metal-rich soils in tropical Africa (Katanga, DR Congo). Ecol. Eng. 82, 81–90 (2015). g p
y
p
(
g ,
g )
g
,
(
)
14. Testiati, E. et al. Trace metal and metalloid contamination levels in soils and in two native plant species of a former industrial site:
E
l
i
f h
h
bili
i
i l J H
d M
248 249 131 141 (2013) g p
y
p
(
g ,
g )
g
,
(
)
14. Testiati, E. et al. Trace metal and metalloid contamination levels in soils and in two native plant species of a former industrial site:
Evaluation of the phytostabilization potential. J. Hazard. Mater. www.nature.com/scientificreports/ 248–249, 131–141 (2013).f 14. Testiati, E. et al. Trace metal and metalloid contamination levels in soils and in two native plant species of a for
Evaluation of the phytostabilization potential. J. Hazard. Mater. 248–249, 131–141 (2013).f phytostabilization potential. J. Hazard. Mater. 248–249, 131–141 5. Cabrera, F., Clemente, L., Díaz Barrientos, E., López, R. & Murillo, J. M. Heavy metal pollution of soils affected by the Guadiamar
toxic fiood. Sci. Total Environ. 242, 117–129 (1999). i
(
)
6. Imperato, M. et al. Spatial distribution of heavy metals in urban soils of Naples city (Italy). Environ. Pollut. 124, 247–256 (2003). i
16. Imperato, M. et al. Spatial distribution of heavy metals in u 7. Gallagher, F. J., Pechmann, I., Bogden, J. D., Grabosky, J. & Weis, P. Soil metal concentrations and vegetative assemblage structure
in an urban brownfield. Environ. Pollut. 153, 351–361 (2008).i i
8. Gallagher, F. J., Pechmann, I., Holzapfel, C. & Grabosky, J. Altered vegetative assemblage trajectories within an urban brownfield
Environ. Pollut. 159, 1159–1166 (2011). 19. Heckenroth, A. et al. Selection of native plants with phytoremediation potential for highly contaminated Mediterranean soil
restoration: Tools for a non-destructive and integrative approach. J. Environ. Manag. 183, 850–863 (2016). 20. Dickinson, N. M., Turner, A. P. & Lepp, N. W. How do trees and other long-lived plants survive in polluted environments?. F
Ecol. 5, 5 (1991).h (
)
21. Partida-Martínez, L. P. & Heil, M. The microbe-free plant: Fact or artifact?. Front. Plant Sci. 2, 100 (2011). h
22. Giller, K. E., Witter, E. & Mcgrath, S. P. Toxicity of heavy metals to microorganisms and microbial processes in agricultural soils:
A review. Soil Biol. Biochem. 30, 1389–1414 (1998). 23. Kabata-Pendias, A. & Pendias, H. Trace Elements in Soils and Plants (CRC Press, 2001). 24. Tyler, G. Heavy metal pollution and mineralisation of nitrogen in forest soils. Nature 255, 701–702 (1975). 25. Seshadri, B., Bolan, N. S. & Naidu, R. Rhizosphere-induced heavy metal(loid) transformation in relation to bioavailability and
remediation. J. Soil Sci. Plant Nutr. https://doi.org/10.4067/S0718-95162015005000043 (2015). 6. Kidd, P. et al. Trace element behaviour at the root–soil interface: Implications in phytoremediation. Environ. Exp. Bot. 67, 243–259
(2009).f 7. Rivera-Becerril, F. Cadmium accumulation and buffering of cadmium-induced stress by arbuscular mycorrhiza in three Pisum
sativum L. genotypes. J. Exp. Bot. 53, 1177–1185 (2002). g
yp
p
(
)
28. Krupa, P. & Kozdrój, J. www.nature.com/scientificreports/ et al. Gain in biodiversity but not in phytostabilization after 3 years of ecological restoration of contaminated Mediter-
ranean soils. Ecol. Eng. 157, 105998 (2020). g
0. Michelaki, C. et al. An integrated phenotypic trait-network in thermo-Mediterranean vegetation describing alternative, coexisting
resource-use strategies. Sci. Total Environ. 672, 583–592 (2019).fhfi g
1. Affholder, M.-C. et al. Transfer of metals and metalloids from soil to shoots in wild rosemary (Rosmarinus officinalis L.) growing
on a former lead smelter site: Human exposure risk. Sci. Total Environ. 454–455, 219–229 (2013).fh 2. Affholder, M.-C. et al. As, Pb, Sb, and Zn transfer from soil to root of wild rosemary: Do native symbionts matter?. Plant Soil 382
219–236 (2014). 3. Ellili, A. et al. Decision-making criteria for plant-species selection for phytostabilization: Issues of biodiversity and functionality
J. Environ. Manag. 201, 215–226 (2017).f g
74. Laffont-Schwob, I. et al. Insights on metal-tolerance and symbionts of the rare species Astraga
bilization of polluted soils and plant conservation. ecmed 37, 57–62 (2011). 4. Laffont-Schwob, I. et al. Insights on metal-tolerance and symbionts of the rare species Astragalus tragacantha aiming at phytosta-
bilization of polluted soils and plant conservation. ecmed 37, 57–62 (2011). 5. Rabier, J. et al. Heavy metal and arsenic resistance of the halophyte Atriplex halimus L. along a gradient of contamination in a
French Mediterranean spray zone. Water Air Soil Pollut. 225, 1993 (2014).i ench Mediterranean spray zone. Water Air Soil Pollut. 225, 1993 (2 p y
6. Quevauviller, Ph. et al. Interlaboratory comparison of EDTA and DTPA procedures prior to certification of extractable trace ele-
ments in calcareous soil. Sci. Total Environ. 178, 127–132 (1996). 7. Anderson, J. P. E. & Domsch, K. H. A physiological method for the quantitative measurement of microbial biomass in soils. Soi
Biol. Biochem. 10, 215–221 (1978). 8 R D
l
t C
T
df 78. R Development Core Team.pdf. 78. R Development Core Team.pdf. 78. R Development Core Team.pdf. 79. Dray, S., Dufour, A. B. & Chessel, D. The ade4 package—II: Two-table and K-table methods. R News 7, 6 (2007). www.nature.com/scientificreports/ & Pajuelo, E. “In situ” phytostabilisation of heavy metal polluted soils using
Lupinus luteus inoculated with metal resistant plant-growth promoting rhizobacteria. J. Hazard. Mater. 177, 323–330 (2010).h 9. Dary, M., Chamber-Pérez, M. A., Palomares, A. J. & Pajuelo, E. “In situ” phytostabilisation of heavy metal polluted soils using
Lupinus luteus inoculated with metal resistant plant-growth promoting rhizobacteria. J. Hazard. Mater. 177, 323–330 (2010). 0. Reichman, S. M. The potential use of the legume–rhizobium symbiosis for the remediation of arsenic contaminated sites. Soil Biol
Bi
h
39 2587 2593 (2007) 60. Reichman, S. M. The potential use of the legume–rhizobium symbiosis for the remediation of arsenic contaminated sites. Soil
Biochem. 39, 2587–2593 (2007). (
)
61. Parraga-Aguado, I., Querejeta, J.-I., González-Alcaraz, M.-N., Jiménez-Cárceles, F. J. & Conesa, H. M. Usefulness of pioneer
vegetation for the phytomanagement of metal(loid)s enriched tailings: Grasses vs. shrubs vs. trees. J. Environ. Manag. 133, 51–58
(2014). (
)
62. Jones, C. G., Lawton, J. H. & Shachak, M. Organisms as ecosystem engineers. Oikos 69, 373 (1994).fi G., Lawton, J. H. & Shachak, M. Organisms as ecosystem enginee 63. Carrasco, L., Azcón, R., Kohler, J., Roldán, A. & Caravaca, F. Comparative effects of native filamentous and arbuscular mycorrhizal
fungi in the establishment of an autochthonous, leguminous shrub growing in a metal-contaminated soil. Sci. Total Environ. 409,
1205–1209 (2011). (
)
64. Padilla, F. M., Ortega, R., Sánchez, J. & Pugnaire, F. I. Rethinking species selection for restoration of arid shrublands. Basic A
Ecol. 10, 640–647 (2009). 65. Ilunga wa Ilunga, E. et al. Plant functional traits as a promising tool for the ecological restoration of degraded tropical metal-rich
habitats and revegetation of metal-rich bare soils: A case study in copper vegetation of Katanga, DRC. Ecol. Eng. 82, 214–221
(2015). (
)
66. Salducci, M.-D. et al. How can a rare protected plant cope with the metal and metalloid soil pollution resulting from past industrial
activities? Phytometabolites, antioxidant activities and root symbiosis involved in the metal tolerance of Astragalus tragacantha. Chemosphere 217, 887–896 (2019). p
67. Kachout, S. S. et al. Accumulation of Cu, Pb, Ni and Zn in the halophyte plant Atriplex grown on polluted soil. J. Sci. Food Agric. 92, 336–342 (2012).fh 68. Schaeffer, A. et al. The impact of chemical pollution on the resilience of soils under multiple stresses: A conceptual framework for
future research. Sci. Total Environ. 568, 1076–1085 (2016).t 9. Tosini, L. www.nature.com/scientificreports/ www.nature.com/scientificreports/ 50. Dias, T. et al. Ammonium as a driving force of plant diversity and ecosystem functioning: Observations based on 5 years’ manipula-
tion of n dose and form in a Mediterranean ecosystem. PLoS ONE 9, e92517 (2014).i y
51. Remon, E. et al. Soil characteristics, heavy metal availability and vegetation recovery at a former metallurgical landfill: Implica
in risk assessment and site restoration. Environ. Pollut. 137, 316–323 (2005). y
y
g
sk assessment and site restoration. Environ. Pollut. 137, 316–323 ( 2. Baumberger, T. et al. Plant community changes as ecological indicator of seabird colonies’ impacts on Mediterranean Islands. Ecol
Ind. 15, 76–84 (2012).f 3. Navas, M.-L., Roumet, C., Bellmann, A., Laurent, G. & Garnier, E. Suites of plant traits in species from different stages of a Medi-
terranean secondary succession: Plant traits and succession. Plant Biol. 12, 183–196 (2010).f 53. Navas, M.-L., Roumet, C., Bellmann, A., Laurent, G. & Garnier, E. Suites of plant traits in species from different stages of a Medi-
terranean secondary succession: Plant traits and succession. Plant Biol. 12, 183–196 (2010). 54 Guillamot, F, Calvert, V, Millot, M -V & Criquet, S Does antimony affect microbial respiration in Mediterranean soils? A micro- f
terranean secondary succession: Plant traits and succession. Plant Biol. 12, 183–196 (2010). 54. Guillamot, F., Calvert, V., Millot, M.-V. & Criquet, S. Does antimony affect microbial respiration in Mediterranean soils? A micro-
cosm experiment Pedobiologia 57 119 121 (2014) 4. Guillamot, F., Calvert, V., Millot, M.-V. & Criquet, S. Does antimony affect microbial respiration in Mediterranean soils? A micro-
cosm experiment. Pedobiologia 57, 119–121 (2014).f p
g
55. Wang, A., He, M., Ouyang, W., Lin, C. & Liu, X. Effects of antimony (III/V) on microbial activities and bacterial community
structure in soil. Sci. Total Environ. 789, 148073 (2021). 6. Oleńska, E. et al. Trifolium repens-associated bacteria as a potential tool to facilitate phytostabilization of zinc and lead polluted
waste heaps. Plants 9, 1002 (2020).f 57. Stambulska, U. Y., Bayliak, M. M. & Lushchak, V. I. Chromium(VI) toxicity in legume plants: Modulation effects of rhizobial
symbiosis. BioMed Res. Int. 2018, 1–13 (2018). y
(
)
58. Karthika, K. S., Rashmi, I. & Parvathi, M. S. Biological functions, uptake and transport of essential nutrients in relation
to plant growth. In Plant Nutrients and Abiotic Stress Tolerance 1–49 (Springer Singapore, 2018). https://doi.org/10.1007/
978-981-10-9044-8_1. 59. Dary, M., Chamber-Pérez, M. A., Palomares, A. J. www.nature.com/scientificreports/ Diffuse and widespread present-day pollution. In Pollution of Marseille’s industrial Calanques—The Impact
of the Past on the Future 204–249 (REF2C, 2016). f
47. Gelly, R. et al. Lead, zinc, and copper redistributions in soils along a deposition gradient from emissions of a Pb-Ag smelter decom-
missioned 100 years ago. Sci. Total Environ. 665, 502–512 (2019).i y
g
48. Tóth, G. et al. Soils of the European Union. JRC Scientific and Technical Reports 85 (2008). i
49. IUSS Working Group WRB. Base de référence mondiale pour les ressources en sols 2014, Mise à jour 2015. Système international de
classification des sols pour nommer les sols et élaborer des légendes de cartes pédologiques. Rapport sur les ressources en sols du monde. Vol. 106 (2015). https://doi.org/10.1038/s41598-022-14139-4 Scientific Reports | (2022) 12:10033 | Author contributions A.H., P.P., H.F., J.R. and I.L.S. contributed to the study conception and design. Material preparation, data col-
lection and analysis were performed by A.H., P.P., H.F., J.R., S.C., M.D.S., L.V. and I.L.S. The first draft of the
manuscript was written by A.H. and all authors commented on previous versions of the manuscript. All authors
read and approved the final manuscript. Funding acquisition was achieved by A.H., H.F. and I.L.S. Competing interests h The authors declare no competing interests. Acknowledgementsh g
This work has been carried out thanks to the support of the Marséco project (ANR Marséco 2008 CESA 018)
and the A*MIDEX project (n° ANR-11-IDEX-0001-02), funded by the Investissements d’Avenir French Govern-
ment programme, managed by the French National Research Agency (ANR). This research was supported by a
CIFRE grant (no. 2012/0175) for Alma Heckenroth PhD, from the Association Nationale de la Recherche et de
la Technologie and the ORTEC Générale de Dépollution company. Many thanks to the Calanques National Park,
particularly to Lidwine Le Mire-Pecheux, for advice and authorization for plant and soil harvesting (authoriza-
tion: di_2015_046_prlv-sol_coronilla-juncea_lafont_imbe). Many thanks to OT-MED scientific network for
improving reflection about global change in the context of management of polluted Mediterranean soils. A. Heckenroth and H. Folzer thank the Fondation ECCOREV for funding the ECCOREV RISQ 2018 project on
crossed risks of pollution and fire in the Calanques hills. Many thanks to Carine Demelas and Maxime Verlande
for their technical help and to Jean-François Mauffrey who did the internal review of this paper with valuable
suggestions. https://doi.org/10.1038/s41598-022-14139-4 Scientific Reports | (2022) 12:10033 | www.nature.com/scientificreports/ Fundingh g
This work has been carried out thanks to the support of the Marséco project (ANR Marséco 2008 CESA 018)
and the A*MIDEX project (n° ANR-11-IDEX-0001-02), funded by the Investissements d’Avenir French Govern-
ment programme, managed by the French National Research Agency (ANR). This research was supported by
a CIFRE grant (no. 2012/0175) for Alma Heckenroth PhD, from the Association Nationale de la Recherche et
de la Technologie and the ORTEC Générale de Dépollution company. A. Heckenroth and H. Folzer thank the
Fondation ECCOREV for funding the ECCOREV RISQ 2018 project on crossed risks of pollution and fire in
the Calanques hills. © The Author(s) 2022 Additional information Correspondence and requests for materials should be addressed to I.L.-S. Reprints and permissions information is available at www.nature.com/reprints. Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and
institutional affiliations. Open Access This article is licensed under a Creative Commons Attribution 4.0 International
License, which permits use, sharing, adaptation, distribution and reproduction in any medium or
format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the
Creative Commons licence, and indicate if changes were made. The images or other third party material in this
article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the
material. If material is not included in the article’s Creative Commons licence and your intended use is not
permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from
the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. © The Author(s) 2022 https://doi.org/10.1038/s41598-022-14139-4 Scientific Reports | (2022) 12:10033 |
|
https://openalex.org/W3004542971
|
https://ieeexplore.ieee.org/ielx7/6287639/8948470/08984362.pdf
|
English
| null |
An Improved LCL-L Compensation Topology for Capacitive Power Transfer in Electric Vehicle Charging
|
IEEE access
| 2,020
|
cc-by
| 9,642
|
Received January 17, 2020, accepted February 3, 2020, date of publication February 5, 2020, date of current version February 14, 2020. Received January 17, 2020, accepted February 3, 2020, date of publication February 5, 2020, date of current version February 14, 2020. Received January 17, 2020, accepted February 3, 2020, date of publication February 5, 2020, date of current version February 14, 2020. Digital Object Identifier 10.1109/ACCESS.2020.2971961 VAN-BINH VU
, (Student Member, IEEE), MOHAMED DAHIDAH
, (Senior Member, IEEE),
VOLKER PICKERT
, (Member, IEEE), AND VAN-TUNG PHAN
, (Senior Member, IEEE)
School of Engineering, Newcastle University, Newcastle upon Tyne NE1 7RU, U.K.
Corresponding author: Van-Binh Vu (v.b.vu2@ncl.ac.uk)
This work was funded by Newcastle University in the Newcastle-Singapore Research Scholarship scheme. VAN-BINH VU
, (Student Member, IEEE), MOHAMED DAHIDAH
, (Senior Member, IEEE),
VOLKER PICKERT
, (Member, IEEE), AND VAN-TUNG PHAN
, (Senior Member, IEEE)
School of Engineering, Newcastle University, Newcastle upon Tyne NE1 7RU, U.K. Corresponding author: Van-Binh Vu (v.b.vu2@ncl.ac.uk)
This work was funded by Newcastle University in the Newcastle-Singapore Research Scholarship scheme. VAN-BINH VU
, (Student Member, IEEE), MOHAMED DAHIDAH
, (Senior Member, IEEE),
VOLKER PICKERT
, (Member, IEEE), AND VAN-TUNG PHAN
, (Senior Member, IEEE)
School of Engineering, Newcastle University, Newcastle upon Tyne NE1 7RU, U.K. Corresponding author: Van-Binh Vu (v.b.vu2@ncl.ac.uk)
This work was funded by Newcastle University in the Newcastle-Singapore Research Scholarship scheme. ABSTRACT This paper proposes an LCL-L compensation circuit for high power capacitive power trans-
fer (CPT) aiming at minimizing number of resonant components and improving system performance. The
proposed topology adopts only four external resonant components at both sides of the capacitive coupler. The output power is proportional to the capacitive coupling coefficient, therefore, it simplifies the design
procedure and abolishes protection circuit requirements under coupler’s misalignment. Moreover, optimizing
efficiency at full-load conditions of compensation network can be easily achieved in this system by designing
resonant components at the highest value of the mutual capacitance. Theoretical analysis of the proposed
system is conducted alongside comparison to lasted CPT compensation circuits. Simulation and experimental
results of a 1.5-kW CPT prototype with an air gap distance of 150 mm are provided to verify the feasibility
and the effectiveness of the proposed system. System performances under different coupler’s misalignment
conditions and output power levels are also examined and discussed as well. INDEX TERMS Capacitive power transfer, compensation circuits, LCL-L topology, electric vehicles, battery
charger. INDEX TERMS Capacitive power transfer, compensation circuits, LCL-L topology, electric vehicles, battery
charger. The associate editor coordinating the review of this manuscript and
approving it for publication was Mohammad S. Khan
. This work is licensed under a Creative Commons Attribution 4.0 License. For more information, see http://creativecommons.org/licenses/by/4.0 I. INTRODUCTION Secondly, the output power is
proportional to capacitive coupling coefficient, which brings
several benefits in terms of design and protection aspects. Dif-
ferentiating from the existing work [7], in which the authors
have only focused on design of the compensation network
for the movable devices with low output power and small air-
gap, a design guideline of a LCL-L compensation network is
given in this paper for the stationary electric vehicle battery
applications with high output power (i.e. 1.5 kW) and large
air-gap (i.e. 15 cm). The third method focuses on the resonant network by
utilizing new compensation topology to upgrade the output
power under low mutual capacitance conditions [21]–[25]. These papers proposed adding extra capacitors connected in
parallel with the capacitive coupler at both sides to solve the
low coupling capacitance problem. Combining with recent
advancement in power semiconductor devices, in which high
frequency and high-power switches are available, the CPT
compensation circuit can be realized at higher than 1 MHz
switching frequency. In [21], a double-sided LCLC topol-
ogy is proposed to transfer 3 kW power with 15 cm air-
gap and achieving 90.8% efficiency. As mentioned in [21],
an additional 100-pF capacitor was connected in parallel with
231-µH series inductor at both sides to operate at 1 MHz of
switching frequency. However, there are eight external com-
ponents in the compensation circuit, reducing the system’s
compactness and increasing the implementation cost. In order
to reduce the number of components and coupler’s installa-
tion space, a four-plate structure for the capacitive coupler
is proposed in [22]. The system in [22] used two separated
pairs of metal, which are vertically integrated to reduce the
space. Moreover, two extra parallel capacitors in [21] can be
replaced by adopting two large coupling capacitors of two
plates that are on the same side in [22]. Double-sided LCLC
topology in [21] can be simplified as double-sided LCL
in [22] with only six external components. To further reduce
the number of components, double-sided LC topology [23],
[24] is proposed by developing from a series compensation
circuit where only an external inductor and capacitor are The theoretical analysis is provided, and relevant equations
are derived to achieve the above features. The relationship
between the output voltage, power and coupling coefficient
is presented, where conditions to attain near Zero-Phase-
Angle (ZPA) and Zero Voltage Switching (ZVS) conditions
of primary inverter are investigated. I. INTRODUCTION Firstly, the highest efficiency point is normally
preferred at the full load condition in power converter system
to reduce total losses [26]. Hence, the CPT system can attain
the highest efficiency by designing compensation circuit of
CPT system at full load and highest value of k, meaning
at perfectly alignment case. Secondly, overpower protection
circuit is not required in any misalignment case, as output
power is also degraded when k decreases. However, the out-
put power in the case of double-sided LC topology [23], [24]
is reversely proportional with k and it is only a function of
the switching frequency and the capacitive coupler value. Consequently, it is very difficult to a design compensation
network for achieving high efficiency at the full load condi-
tion and furthermore, the protection requirement is a must if
misalignment occurs. This also reduces flexibility to design
the capacitive coupler for attaining a relatively large coupling
coefficient resulting in better system efficiency. FIGURE 1. A capacitive charger system for electric vehicles. In order to address the above issues, several solutions
have been proposed recently. The first solution utilizes wide-
bandgap semiconductors (SiC/GaN) to operate the CPT
charger at high frequency for compensating the low mutual
capacitance and achieving high power density [17], [18]. Most of the recent CPT systems operate at a frequency of
greater than 1 MHz. In [18], the new design method of the
6.78-MHz CPT system is proposed to mitigate the negative
effect of parasitic elements. The second approach uses the
multi-modular architecture to achieve high power transfer
while maintaining the safety limits [19], [20]. Therefore, the
total output power is summed up from each module’s power. However, this approach increases the component counts con-
sisting of power electronics converters as well as coupling
capacitor plates and resonant networks. This paper introduces the LCL-L compensation circuit
topology (i.e. illustrated in Fig. 2) for high power and large
air-gap CPT systems such as Electric Vehicle battery charging
to eliminate the abovementioned drawbacks associated with
double-sided LC topology. The first feature of the proposed
topology is that only four external components are required,
which is similar to the topology presented in [23], [24]. An LCL compensation network is connected at the primary
side and an inductor is connected in series with capacitive
coupler at the secondary side. I. INTRODUCTION high voltage stresses among the capacitor coupler plates and
compensation components. To simplify the capacitive cou-
pler structure, the CPT systems with only two coupling plates
are proposed in [12], [13]. The output power in several CPT
systems can be increased to kilowatt scales for applications
such as contactless stationary EVs charging [14], [15] or
dynamic EVs charging [16]. As compared with an Inductive Power Transfer (IPT) sys-
tem, a capacitive power transfer (CPT) system has several
benefits such as implementing low-cost and insensitive to
nearby metal objects [1]. The developments of contactless
CPT systems for low power and low air-gap applications
such as consumer electronics, LED lighting, battery charging
have been received the most attention [2]–[13]. The single
series inductor is normally added into either one side or both
sides of capacitor plates to compensate for the high leakage
capacitances [2]–[4]. The CPT system and the associated
compensation circuits for multiple pickups constant current
output applications are presented in [5]. To prevent the elec-
trical and health hazards when the secondary side load is
removed suddenly, the different design approaches of the
compensation networks are discussed in [6]–[8]. In addition,
different methods are introduced in [9], [10] to reduce the However, the CPT technology has still not been widely
used in high power and large air-gap applications due to
its limitation of low mutual capacitance. When the air-
gap reaches to around 15-20 cm for applications such as
Electric Vehicle battery charging (i.e. illustrated in Fig. 1),
then mutual capacitance values between two coupling plates
reduces to picofarad range [21]. Consequently, it requires
very large inductors in the compensation circuit if simple
series compensation topologies are used [2], [3]. Low mutual
capacitance also results in very large voltage stresses on
resonant inductors. In terms of energy conversion, high effi-
ciency is very difficult to achieve at low coupling capacitance
condition. The associate editor coordinating the review of this manuscript and
approving it for publication was Mohammad S. Khan
. 27757 V.-B. Vu et al.: Improved LCL-L Compensation Topology for Capacitive Power Transfer in Electric Vehicle Charging connected in series and parallel with the capacitive coupler
in each side respectively. FIGURE 1. A capacitive charger system for electric vehicles. In both [21] and [22], the output power is proportional to
the capacitive coupling coefficient k, which brings several
advantages. II. PROPOSED LCL-L COMPENSATION
TOPOLOGY CPT SYSTEM
A. OPERATING PRINCIPLE At the secondary side, the voltage loop can be expressed
by Eqs (3-4). Figure 2a illustrates the proposed LCL-L compensation
topology for CPT, which comprises a full-bridge inverter
at the primary side, compensation networks in both sides,
capacitive coupler with integrated structure [22] and a bridge
rectifier at the secondary side. CPT system is energized
by a DC Voltage source Vin to provide a DC power for
an output battery load, which in this case is simply repre-
sented by resistive load, Ro, for simplicity. Vo and Io are the
required output voltage and output current, respectively. In this paper, the First Harmonic Approximation (FHA) is
used to analyze the proposed system in the frequency-domain
for simplicity. Therefore, first harmonic components can be
derived from the equivalent circuit of Fig. 3 as given by (1). Therefore, vin_AC, vo_AC and io_AC in (1) are the first harmonic
components of the input voltage, output voltage and current
of the compensation circuit, respectively. In the following
analysis Vin, V1, V2, Vo, Iin, I1, and Io are used to express
the complex form of the corresponding variables. Io = jωCMV1 −jωC2V2
(3)
Vo = V2 −jωL2Io
=
1 −ω2L2C2
V2 + ω2L2CMV1
(4)
ω2L2C2 = 0
(5) (3) 1 −ω2L2C2 = 0 (5) In order to simplify the control of Vo, L2 and C2 should
form a series resonance as shown in (5). Therefore, the output
voltage directly depends on the mutual capacitance between
the two sides, CM and the voltage value on equivalent pri-
mary capacitor V1. Moreover, V1 in the CPT system plays
a similar role as the transmitter coil current in the IPT sys-
tem [27]. Similarly, to simplify the control of output voltage,
the value of V1 should be maintained constant regardless of
load condition at the secondary side. I. INTRODUCTION A flowchart of the
detailed design procedure is illustrated alongside with sim-
ulation results. Comparison is presented between doubled-
side LCLC presented in [21], [22], doubled-side LC reported
in [23], [24] and the proposed LCL-L topologies, consider-
ing the number of external components and output power
characteristics. A laboratory prototype of 1.5-kW CPT is
developed to experimentally validate the proposed topol-
ogy. The performance of the proposed circuit topology is
further examined under misalignment and different load
conditions. 27758 VOLUME 8, 2020 VOLUME 8, 2020 V.-B. Vu et al.: Improved LCL-L Compensation Topology for Capacitive Power Transfer in Electric Vehicle Charging FIGURE 2. (a) The proposed LCL-L topology for capacitive power transfer, (b) Structure of the capacitive coupler. FIGURE 2. (a) The proposed LCL-L topology for capacitive power transfer, (b) Structure of the capacitive coupler. FIGURE 3. Equivalent circuit model of LCL-L compensation topology. for capacitive coupler. The value of C1, C2 and CM are
depicted in (2) according to [22]. C1 = C12 + (C13 + C14) (C23 + C24)
C13 + C14 + C23 + C24
C2 = C34 + (C13 + C23) (C14 + C24)
C13 + C14 + C23 + C24
CM =
C24C13 −C14C23
C13 + C14 + C23 + C24
(2 FIGURE 3. Equivalent circuit model of LCL-L compensation topology. (2) VOLUME 8, 2020 B. COMPARISON BETWEEN CPT COMPENSATION
TOPOLOGIES The input impedance for CPT system Zin can be derived
from the equivalent circuit depicted in Fig. 2 in the same way
as the IPT system, where the phase angle of Zin indicates the
phase difference between Vin and Iin. To minimize the losses
and stresses on the primary inverter, the phase angle of Zin
should be positive and close to zero to achieve soft-switching
turn-on for MOSFETs of the primary inverter [22]. This
section explores the conditions to attain Zero-Phase-Angle
(ZPA) of Zin. Fig. 3 shows the simplified model of Fig 2 where
ZS represents the secondary circuit’s impedance and ZR is the
reflection of ZS to the primary side as expressed in (13). In this section, doubled-side LCLC, doubled-side LC, and
the proposed LCL-L topologies are compared to each other,
considering the following aspects: (a) the number of external
components (b) output power characteristic (c) efficiency at
rated power and (d) design flexibility. This is summarized
in Table 1 below. As demonstrated by Table 1, minimiz-
ing the number of components is an important feature of
double-sided LC and the proposed LCL-L topology in which
each topology only requires four external components, in
addition, the coupling capacitors. Smaller number of compo-
nents, lower implementation cost, and higher power density. The second factor is the output power, which is inversely
proportional to the mutual capacitance Cm in the case of
double-sided LC topology while it is proportional to Cm
for both, the double-sided LCLC and the proposed LCL-L. Design compensation circuits necessitate that the highest
value of Po must correspond to the highest value of Cm. This
is to guarantee that maximum efficiency can be attained at
the highest output power point. On the other hand, when
misalignment occurs, Cm drops below certain range and the
input power exceeds the limited value, therefore overpower
protection is essential for double-sided LC to stop charger
while double-sided LCLC and the proposed topology do not
require one as output power is proportional with Cm. ZS =
V2
jωCMV1
=
1
jωC2 (jωL2 + Rac)
1
jωC2 + jωL2 + Rac
(13)
ZR =
V1
−jωCMV2
=
1
ω2C2
MZS
(14) (13) (14) Therefore, the input impedance of the proposed system, Zin
can then be written as (15), considering the resonant operation
condition described by (5) and (9). II. PROPOSED LCL-L COMPENSATION
TOPOLOGY CPT SYSTEM
A. OPERATING PRINCIPLE jωL1 +
1
jωC1
+
1
jωC1p
= 0
(16)
Zin =
1
1 −ω2L1C1p
C2
M
ω2L2
2 +R2ac
×
h
ω2C1C2L1pL2Rac+jωL1p
ω2L2
2C2
M + C1C2R2
ac
i
(17) I1 = −jωC1pVin =
1
jωL1p
Vin
(8)
jωL1p +
1
jωC1p
= 0
(9) (16) (8) (9) From (6), to keep V1 independent of load condition, then
1/(ωC1p) should be very small as compared to ωL1. Eq. (10)
expresses V1 under this constraint combining with (8). From (6), to keep V1 independent of load condition, then
1/(ωC1p) should be very small as compared to ωL1. Eq. (10)
expresses V1 under this constraint combining with (8). i
(17) Hence, the phase angle of Zin can be calculated as follows: V1 ≈−jωL1I1 = −L1
L1p
Vin
(10) ϕ (Zin)=tan−1
Im (Zin)
Re (Zin)
=tan−1
"
ω2L2
2C2
M + C1C2R2
ac
ωC1C2L2Rac
#
(18) (10) Therefore, the output voltage of the proposed topology is
independent of the load condition and is proportional with a
coupling capacitor between the two sides. Eqs. (11) and (12)
depict Vo and output power Po while Io is the output battery
current, which is a function of the load condition. Conse-
quently, Po is also proportional to the coupling capacitor CM. It is noted that the phase of output voltage Vo and input
voltage Vin are reversed. In other words, the phase-shift θ in
(1) between Vo and Vin is 180o. #
(18) The design procedure in this paper neglects the term
ω2L2
2C2
M as compared to C1C2R2
ac. Therefore, ϕ (Zin) ≈
tan−1
Rac
ωL2
, this can be noticeable that ϕ (Zin) is always
positive. MOSFETs of the primary inverter are turned on at
a zero-voltage switching (ZVS) condition. Furthermore, Rac
can be calculated according to load requirement while the
switching frequency can be maintained at around 1-MHz. Therefore, L2 should be selected for ωL2 to be large enough
as compared to Rac. This constraint guarantees the near ZPA
operation for the primary inverter. Vo ≈−ω2L2CM
L1
L1p
Vin
(11)
Po = VoIo ≈ω2L2CM
L1
L1p
VinIo
(12) (11) (12) II. PROPOSED LCL-L COMPENSATION
TOPOLOGY CPT SYSTEM
A. OPERATING PRINCIPLE At the primary side, the
relationship between Vin and V1 can be expressed as follows: V1 = VC1p + VL1 = (I1 −Iin)
1
jωC1p
−jωL1I1
=
1
jωC1p
−jωL1
I1 −
1
jωC1p
Iin
(6)
Vin = jωL1pIin + (Iin −I1)
1
jωC1p
=
jωL1p +
1
jωC1p
Iin −
1
jωC1p
I1
(7)
vin_AC (t) = 4
π Vin sin (2πft)
vo_AC (t) = 4
π Vo sin (2πft + θ)
io_AC (t) = π
2 Io sin (2πft + θ)
RAC = 8
π2
Vo
Io
= 8
π2 Ro
(1) (6) (1) (7) As it can be noticed from Fig. 2 (a) and (b), the capacitor
between plates Pi and Pj is given by Cij (i, j = 1,4). The
coupling capacitors C12 and C34 are adopted to form two extra
parallel capacitors, which use to boost the voltage supplied where VC1p and VL1 are voltage on C1p and L1 respectively. Similar to the primary LCC topology in IPT systems [28],
the L1p and C1p form a resonant circuit and function as an
input filter, which removes high-order harmonics from the where VC1p and VL1 are voltage on C1p and L1 respectively. where VC1p and VL1 are voltage on C1p and L1 respectively. Similar to the primary LCC topology in IPT systems [28],
the L1p and C1p form a resonant circuit and function as an Similar to the primary LCC topology in IPT systems [28],
the L1p and C1p form a resonant circuit and function as an
input filter, which removes high-order harmonics from the VOLUME 8, 2020 V.-B. Vu et al.: Improved LCL-L Compensation Topology for Capacitive Power Transfer in Electric Vehicle Charging is described by (16). Therefore, the simplified version of Zin
is given by (17). current injected to primary plates. Furthermore, their reso-
nance makes I1 constant regardless of the load condition. As a result, I1 can then be derived as (8) with the condition
presented in (9). B. COMPARISON BETWEEN CPT COMPENSATION
TOPOLOGIES In the next step, specifications of the CPT system
are determined, and equivalent load resistance is calculated
based on the charging current and voltage. In the meantime,
the size and geometry of the CPT coupler are determined
based on the air-gap of the coupler and the limitations of the
installation area. Once the coupler’s dimensions are defined,
then C1, C2 and CM can be estimated by using electro-
magnetic field simulation software such as MAXWELL 3D,
while L2 can be calculated by (5). At this point, it is also
necessary to see if the set of parameters can also satisfy the
conditions of Rac ≪ωoL2. If results are not satisfactory, then
C1, C2 and CM need to be adjusted by varying the coupler’s
dimension. Value of CM is the simulated value with 15-cm
air-gap and no misalignment. The FEA simulation model and
final dimensions of the capacitive coupler can be found in
Fig. 6 in which the dimensions of plates P1 and P2 are iden-
tical with P3 and P4, respectively. The thickness of all four
plates are the same as 2 mm. Once all the above conditions
are matched, then values of L1, L1p and C1p are calculated the system, resulting in lower efficiencies at heavy loads. In double-sided LCLC and the proposed topology, efficiency
optimization is easier as it only needs to design the highest Po
value when Cm reaches maximum value (no misalignment). However, if misalignment occurs, Cm is reduced with reduc-
tion of Po. Efficiency will be also dropped, however, at a
lower level of output power. Another factor is the system flexibility in designing the
capacitive coupler. As shown in Table 1, the output power
can be regulated by adjusting the ratio between Cf 1Cf 2 and
C1C2 in an LCLC-compensated CPT system. Similarly, in the
proposed topology, the output power can also be adjusted by
varying the values of L1, L2 and L1p when coupling capacitors
are fixed. This feature provides the flexibility to design the
capacitive coupler to achieve a relatively large mutual capac-
itance Cm, hence a better system efficiency. This advantage,
however, is incapable to achieve in double-sided LC topology. III. DESIGN PROCEDURE AND SIMULATION
VERIFICATION B. COMPARISON BETWEEN CPT COMPENSATION
TOPOLOGIES Zin =
jωL1p
1 + jωC1p
jωL1 +
ZR
1+jωC1ZR
=
jωL1p (1 + jωC1ZR)
1 −ω2C2
1p + jωZR
C1 + C1p −ω2C1C1pL1
(15) (15) To further simplify Zin, another resonant loop at the pri-
mary side, between C1, C1p, and L1 should be formed, which Moreover, if the protection circuit is not required, then
Cm must be reduced in order to increase the power level of 27760 VOLUME 8, 2020 VOLUME 8, 2020 V.-B. Vu et al.: Improved LCL-L Compensation Topology for Capacitive Power Transfer in Electric Vehicle Charging TABLE 1. Comparison between different CPT compensation topologies. TABLE 1. Comparison between different CPT compensation topologies. First, the operating frequency needs to be designed by consid-
ering the volume and losses of resonant components as well as
power switches, therefore, the frequency of 1 MHz is selected
in this work. In the next step, specifications of the CPT system
are determined, and equivalent load resistance is calculated
based on the charging current and voltage. In the meantime,
the size and geometry of the CPT coupler are determined
based on the air-gap of the coupler and the limitations of the
installation area. Once the coupler’s dimensions are defined,
then C1, C2 and CM can be estimated by using electro-
magnetic field simulation software such as MAXWELL 3D,
while L2 can be calculated by (5). At this point, it is also
necessary to see if the set of parameters can also satisfy the
conditions of Rac ≪ωoL2. If results are not satisfactory, then
C1, C2 and CM need to be adjusted by varying the coupler’s
dimension. Value of CM is the simulated value with 15-cm
air-gap and no misalignment. The FEA simulation model and
final dimensions of the capacitive coupler can be found in
Fig. 6 in which the dimensions of plates P1 and P2 are iden-
tical with P3 and P4, respectively. The thickness of all four
plates are the same as 2 mm. Once all the above conditions
are matched, then values of L1, L1p and C1p are calculated First, the operating frequency needs to be designed by consid-
ering the volume and losses of resonant components as well as
power switches, therefore, the frequency of 1 MHz is selected
in this work. III. DESIGN PROCEDURE AND SIMULATION
VERIFICATION The flowchart in Fig. 5 illustrates the practical procedure for a
designing compensation circuit for the proposed CPT system. 27761 27761 VOLUME 8, 2020 V.-B. Vu et al.: Improved LCL-L Compensation Topology for Capacitive Power Transfer in Electric Vehicle Charging FIGURE 6. FEA model and dimensions of the capacitive coupler. FIGURE 7. Voltage gain and phase of input impedance of the LCL-L circuit
with parameters in Table 2. 1 MH
M
hil
th
h
l
f Z
t 1 MH
i FIGURE 4. Simplified equivalent circuit of Fig. 3 (a) Primary side,
(b) Secondary side. FIGURE 5. Flowchart of the proposed CPT design procedure. FIGURE 6. FEA model and dimensions of the capacitive coupler. FIGURE 7. Voltage gain and phase of input impedance of the LCL-L circuit
with parameters in Table 2. 1 MHz. Meanwhile, the phase angle of Zin at 1 MHz is
always positive and very close to zero. This confirms that
the proposed topology enables the CPT system to operate
in the Constant Voltage (CV) mode with near ZPA achieve-
ment. This is further confirmed by the simulation results of
different load conditions as illustrated in Fig. 8. Input voltage FIGURE 4. Simplified equivalent circuit of Fig. 3 (a) Primary side,
(b) Secondary side. FIGURE 4. Simplified equivalent circuit of Fig. 3 (a) Primary side,
(b) Secondary side. FIGURE 6. FEA model and dimensions of the capacitive coupler. FIGURE 7. Voltage gain and phase of input impedance of the LCL-L circuit
with parameters in Table 2. FIGURE 6. FEA model and dimensions of the capacitive coupler. FIGURE 6. FEA model and dimensions of the capacitive coupler. FIGURE 6. FEA model and dimensions of the capacitive coupler. FIGURE 4. Simplified equivalent circuit of Fig. 3 (a) Primary side,
(b) Secondary side. FIGURE 5. Flowchart of the proposed CPT design procedure. FIGURE 7. Voltage gain and phase of input impedance of the LCL-L circuit
with parameters in Table 2. 1 MHz. Meanwhile, the phase angle of Zin at 1 MHz is
always positive and very close to zero. This confirms that
the proposed topology enables the CPT system to operate
in the Constant Voltage (CV) mode with near ZPA achieve-
ment. This is further confirmed by the simulation results of
different load conditions as illustrated in Fig. 8. III. DESIGN PROCEDURE AND SIMULATION
VERIFICATION Input voltage
Vin slightly delays with input current Iin and the phase-shift
between them is very small, confirming that the resonant
network achieves near ZPA operation. Soft-switching oper-
ation for the inverter (i.e. ZVS) is also achieved as the cut-off
current at the switching transient is about 5 A. In addition,
the input and output voltage are out of phase as predicted
by (11). Moreover, when output power varies, the shape of FIGURE 5. Flowchart of the proposed CPT design procedure. using (9), (11) and (16). Furthermore, the specifications of
the CPT charger and all other parameters are summarized
in Table 2. In order to verify the above analysis and design
procedure, the voltage gain GV and the phase of the input
impedance Zin are drawn as shown in Fig. 7. Figure 7 shows that the voltage gain is constant regardless
of the load conditions at the resonant frequency of around 27762 27762 VOLUME 8, 2020 VOLUME 8, 2020 V.-B. Vu et al.: Improved LCL-L Compensation Topology for Capacitive Power Transfer in Electric Vehicle Charging FIGURE 8. Simulation results of input waveforms and output waveforms
with Vin = 300 V and when the load at (a) 40 and input waveforms
when the load at (b) 20 and (c) 10 . FIGURE 9. Experimental setup for the proposed LCL-L CPT charger
(a) capacitive coupler (b) The resonant networks in both sides. (IDW30G65C5 from Infineon) are used for the secondary
f ll b id
tifi TABLE 2. CPT system parameters. FIGURE 8. Simulation results of input waveforms and output waveforms
with Vin = 300 V and when the load at (a) 40 and input waveforms
when the load at (b) 20 and (c) 10 . TABLE 2. CPT system parameters. TABLE 2. CPT system parameters. Iin waveform changes accordingly. As the proposal LCL-L
is the voltage source, then resistive load value Ro decreases
when output power increases (i.e. reduce from 40 to 10
in simulation). At the smallest value of Ro of 10 (also
the heaviest load), phase-shift between voltage and current
is smallest and ZPA condition is very close. It also confirms
the above condition of Rac ≪ωoL2. Phase-shift between Vin
and Iin slightly increases when Ro increases as the CPT circuit
goes towards to light load condition. Nevertheless, it is worth
noting from Fig. III. DESIGN PROCEDURE AND SIMULATION
VERIFICATION 8a that the input current waveform, Iin is not
a sinusoidal due to high harmonics. However, when the load
increases then the input current becomes nearly sinusoidal at
the full load (Fig. 8c). The resonance between L1p and C1p
acts as a high-pass filter, which removes higher harmonics
components at light load and provides a sinusoidal voltage
and current for the primary capacitor plates. FIGURE 8. Simulation results of input waveforms and output waveforms
with Vin = 300 V and when the load at (a) 40 and input waveforms
when the load at (b) 20 and (c) 10 . FIGURE 8. Simulation results of input waveforms and output waveforms
with Vin = 300 V and when the load at (a) 40 and input waveforms
when the load at (b) 20 and (c) 10 . FIGURE 9. Experimental setup for the proposed LCL-L CPT charger
(a) capacitive coupler (b) The resonant networks in both sides. IV. EXPERIMENTAL VERIFICATION 10c presents the voltage stresses on the capacitor
coupler at the primary side between P1 and P2 and at the
secondary side between P3 and P4. They are also equivalent
to voltages V1 and V2 in Fig. 3, respectively, where their
phase-shift is of 90 degrees. As the voltage range of probe is
limit, then the waveforms are captured with an input voltage
of only 100V. At full load conditions, these voltage stresses
have RMS values up to 4 and 5.5 kV for primary and sec-
ondary couplers respectively. Therefore, the distance between
plate P1 and P2 and between plates P3 and P4 should be large
enough to avoid arcing between two plates. According to the
breakdown voltage of air value, then the distance between
plates is set at 1 cm for both sides in this work. For completeness, the variations of mutual capacitance
with respect to different conditions of misalignment or
air-gap is also experimentally investigated and depicted
i
Fi
11 It
f
d th t th
l
f C
d
b FIGURE 11. Variation of mutual capacitance CM versus (a) Misalignments. (b) Different air-gap. GURE 10. Experimental waveforms of (a) input voltage and current,
b) output voltage and current at rated load, (c) voltages on primary and
econdary capacitive couplers with Vin = 100 V. imulation results shown in Fig. 8c. At full-load conditio
he output power of 1.5 kW is obtained at the secondary sid
or which the waveforms of output resonant network’s curre FIGURE 11. Variation of mutual capacitance CM versus (a) Misalignments. (b) Different air-gap. Io and voltage Vo are illustrated in Fig. 10b. It should be
noted that as the circuit operates at a very high frequency, then
some small ringing and spikes occurred in the waveforms. However, this can be suppressed or minimized with a better
design of power PCB. Fig. 10c presents the voltage stresses on the capacitor
coupler at the primary side between P1 and P2 and at the
secondary side between P3 and P4. They are also equivalent
to voltages V1 and V2 in Fig. 3, respectively, where their
phase-shift is of 90 degrees. As the voltage range of probe is
limit, then the waveforms are captured with an input voltage
of only 100V. At full load conditions, these voltage stresses
have RMS values up to 4 and 5.5 kV for primary and sec-
ondary couplers respectively. IV. EXPERIMENTAL VERIFICATION Fig. 9 portrays a laboratory prototype, which was developed
based on the system parameters tabulated in Table 2 to vali-
date the simulation and the theoretical findings. As mentioned
earlier, capacitive coupler (i.e. Fig. 9a) consists of four alu-
minum plates with vertical structure. All these four plates are
of square shape with the dimensions of 915 mm × 915 mm
and 615 mm × 615 mm for large and small plates, respec-
tively. Fig. 9b depicts the resonant network part. High-
power and high-frequency polypropylene film capacitors
from TDK are used to construct the resonant capacitor C1p
due to their low equivalent series resistance (ESR) and high
current carrying capability. All resonant inductors are con-
structed using Litz wire of 38 AWG to reduce the conduc-
tion losses at high frequency. To prevent potential arcing
between different inductor’s turns, the PVC isolation tape
is wounded around the copper Litz wire. The full-bridge
inverter is built using low-RdsON Silicon Carbide (SiC) MOS-
FETs (C2M0025120D from Cree) while four SiC diodes FIGURE 9. Experimental setup for the proposed LCL-L CPT charger
(a) capacitive coupler (b) The resonant networks in both sides. (IDW30G65C5 from Infineon) are used for the secondary
full-bridge rectifier. As discussed in Section III, the rated output power is
attained at the highest value of CM when all plates perfectly
aligned with 15-cm airgap. Experimental waveforms of input
resonant network’s voltage and current at the above condition
are depicted in Fig. 10a at Ro = 10 and input DC voltage
of 200 V. It can be noticed that ZPA between Vin and Iin
and the shape of Iin are in a very good agreement with the 27763 27763 VOLUME 8, 2020 V.-B. Vu et al.: Improved LCL-L Compensation Topology for Capacitive Power Transfer in Electric Vehicle Charging FIGURE 10. Experimental waveforms of (a) input voltage and current,
(b) output voltage and current at rated load, (c) voltages on primary and
FIGURE 11. Variation of mutual capacitance CM versus (a) Misalignments. (b) Different air-gap. Io and voltage Vo are illustrated in Fig. 10b. It should be
noted that as the circuit operates at a very high frequency, then
some small ringing and spikes occurred in the waveforms. However, this can be suppressed or minimized with a better
design of power PCB. Fig. IV. EXPERIMENTAL VERIFICATION (b) Different air-gap. FIGURE 13. System’s efficiency when Ro = 10 according different
conditions (a) Misalignments. (b) Different air-gap. CPT system can maintain efficiency above 76% even with
a 30-cm misalignment or 25-cm air-gap. When metal plates
perfectly aligned, then power transfer can be achieved at
the rated value of 1.5-kW with the efficiency of 85.5 %. These results verify that the output power is proportional
with CM and the maximum output power is achieved at the
highest value of CM. As highlighted earlier, this is a beneficial
feature enabling optimizing efficiency at high loads when the
maximum output power is calculated at the highest value
of the mutual capacitance. Furthermore, the output power
cannot exceed the rated value of 1.5-kW if misalignment
or air-gap variation occurs, thus the protection circuit is not
necessary. FIGURE 14. Power loss distribution of the proposed system components. FIGURE 14. Power loss distribution of the proposed system components. The power loss distribution is estimated for the proto-
type based on the loss analysis for every single component. Detailed analysis is given in the APPENDIX. We consider the
losses on the following components: inverter’s switches, L1p,
C1p, L1, capacitive coupler, L2 and rectifier. FIGURE 14. Power loss distribution of the proposed system components. FIGURE 14. Power loss distribution of the proposed system components. After all calculations, the remaining losses belong to the
capacitive aluminium coupler. The loss distribution of the
proposed converter is depicted in Fig. 14 with the total
estimated loss of 218W at rated power. According to that
aluminium coupler and resonant inductor L2 contribute the
highest losses which are 26.67% and 32.5% respectively. It can be explained that with the vertical structure coupler
in Fig. 2b, there is an existing crossing coupling capacitive
(i.e. C12, C14, C23 and C34) which produces higher losses
in the couplers. Moreover, the current on L2 (i.e. same with
output current Io) is near twice the current on L1, therefore
producing double losses in inductor L2 as compared to L1. Other components dissipate less than 18% of the total loss. p
To reduce losses at an operating frequency of 1MHz, for
all the inductors (L1p, L1, L2), the Litz wire (Litz wire-
600 strands-AWG 38) is used while air-core is adopted. Moreover, the capacitor C1p is constructed in parallel from
several low-ESR capacitors from TDK to minimize the total
resistance. IV. EXPERIMENTAL VERIFICATION The power loss distribution is estimated for the proto-
type based on the loss analysis for every single component. Detailed analysis is given in the APPENDIX. We consider the
losses on the following components: inverter’s switches, L1p,
C1p, L1, capacitive coupler, L2 and rectifier. To reduce losses at an operating frequency of 1MHz, for
all the inductors (L1p, L1, L2), the Litz wire (Litz wire-
600 strands-AWG 38) is used while air-core is adopted. Moreover, the capacitor C1p is constructed in parallel from
several low-ESR capacitors from TDK to minimize the total
resistance. The ESR value can be calculated based on the
dissipation factor (DF) (i.e. ESR
=
DF ∗
1
2πfC1p ). For
Inverter switches, the switching loss is insignificant com-
pared with conduction loss due to achieving ZVS turn on. At the secondary side, the loss on the rectifier is calcu-
lated based on the forward voltage of SiC Schottky Diodes. FIGURE 13. System’s efficiency when Ro = 10 according different
conditions (a) Misalignments. (b) Different air-gap. FIGURE 14. Power loss distribution of the proposed system components. After all calculations, the remaining losses belong to the
capacitive aluminium coupler. The loss distribution of the
proposed converter is depicted in Fig. 14 with the total
estimated loss of 218W at rated power. According to that
aluminium coupler and resonant inductor L2 contribute the
highest losses which are 26.67% and 32.5% respectively. It can be explained that with the vertical structure coupler
in Fig. 2b, there is an existing crossing coupling capacitive
(i.e. C12, C14, C23 and C34) which produces higher losses
in the couplers. Moreover, the current on L2 (i.e. same with
output current Io) is near twice the current on L1, therefore
producing double losses in inductor L2 as compared to L1. Other components dissipate less than 18% of the total loss. FIGURE 12. Output power and system’s efficiency when Ro = 10
according different conditions of (a) Misalignments. (b) Different air-gap. FIGURE 13. System’s efficiency when Ro = 10 according different
conditions (a) Misalignments. (b) Different air-gap. FIGURE 12. Output power and system’s efficiency when Ro = 10
according different conditions of (a) Misalignments. (b) Different air-gap. FIGURE 12. Output power and system’s efficiency when Ro = 10
according different conditions of (a) Misalignments. (b) Different air-gap. FIGURE 13. System’s efficiency when Ro = 10 according different
conditions (a) Misalignments. IV. EXPERIMENTAL VERIFICATION Therefore, the distance between
plate P1 and P2 and between plates P3 and P4 should be large
enough to avoid arcing between two plates. According to the
breakdown voltage of air value, then the distance between
plates is set at 1 cm for both sides in this work. For completeness, the variations of mutual capacitance
with respect to different conditions of misalignment or
air-gap is also experimentally investigated and depicted
in Fig. 11. It was found that the value of CM drops by
63% under 30-cm misalignment while reduced by 49%
when the air-gap increases from 15 to 25 cm. As a result,
the output power and system’s efficiency are also decreased
as illustrated in Fig. 12 and 13. However, the proposed FIGURE 10. Experimental waveforms of (a) input voltage and current,
(b) output voltage and current at rated load, (c) voltages on primary and
secondary capacitive couplers with Vin = 100 V. simulation results shown in Fig. 8c. At full-load condition,
the output power of 1.5 kW is obtained at the secondary side
for which the waveforms of output resonant network’s current VOLUME 8, 2020 VOLUME 8, 2020 27764 V.-B. Vu et al.: Improved LCL-L Compensation Topology for Capacitive Power Transfer in Electric Vehicle Charging sfer in Electric Vehicle Charging
FIGURE 13. System’s efficiency when Ro = 10 according different
conditions (a) Misalignments. (b) Different air-gap. FIGURE 14. Power loss distribution of the proposed system components. V.-B. Vu et al.: Improved LCL-L Compensation Topology for Capacitive Power Transfer in Electric Vehicle Charging
FIGURE 12. Output power and system’s efficiency when Ro = 10
according different conditions of (a) Misalignments. (b) Different air-gap. CPT system can maintain efficiency above 76% even with
a 30-cm misalignment or 25-cm air-gap. When metal plates
perfectly aligned, then power transfer can be achieved at
the rated value of 1.5-kW with the efficiency of 85.5 %. These results verify that the output power is proportional
with CM and the maximum output power is achieved at the
highest value of CM. As highlighted earlier, this is a beneficial
feature enabling optimizing efficiency at high loads when the
maximum output power is calculated at the highest value
of the mutual capacitance. Furthermore, the output power
cannot exceed the rated value of 1.5-kW if misalignment
or air-gap variation occurs, thus the protection circuit is not
necessary. V. CONCLUSION An improved LCL-L compensation circuit topology for a
high-power CPT system is presented in this paper. An LCL
compensation network is constructed at the primary side and
an inductor is connected in series with the capacitive coupler
at the secondary side. The proposed design enables achieving
near ZPA operation and ZVS condition for primary inverter
as well as the constant voltage between two primary capac-
itive couplers. Therefore, the output power is proportional
to the coupling capacitance which brings several benefits in
terms of design and overpower protection aspects. Further-
more, the number of external components is also minimized
which helps to improve the system’s compactness and reduce
implementing cost. The proposed topology neglects almost
drawbacks from previous CPT topologies and remains the
good characteristics as mentioned before. Detailed analysis,
design steps, and comparison are provided combining with
simulated and experimental results to validate circuit topol-
ogy. The proposed system can achieve an efficiency of 85.5%
with 1.5-kW output power and a 150-mm air-gap. (b) On inductor L1, where RL1 is the total AC and DC
resistances of L1. I1 is the RMS value of current through the
primary plate. Ploss_L1 = I2
inRL1
(A4) (A4) (c) On capacitor C1p, where RC1p is the total AC and DC
resistances of C1p. Ploss_C1p = I2
C1pRC1p
(A5) (A5) (d) On inductor L2, where RL2 is the total AC and DC
resistances of L1. I1 is the RMS value of current through the
primary plate. Ploss_L1p = I2
1 RL2
(A6) (A6) All four resistance values of RL1p, RL1, RC1p and RL1 can
be estimated easily based on datasheets of Litz wire and
capacitor. In experiments, these values have measured again
by using the RLC meter. (4) Conduction losses on rectifier: in the proposed system,
secondary rectifier is built from SiC Schottky Diodes. The
loss on the rectifier can be found in (A7) where VfwD and
rD are the forward voltage and the On-State resistance of
rectifier’s diode. IV. EXPERIMENTAL VERIFICATION The ESR value can be calculated based on the
dissipation factor (DF) (i.e. ESR
=
DF ∗
1
2πfC1p ). For
Inverter switches, the switching loss is insignificant com-
pared with conduction loss due to achieving ZVS turn on. At the secondary side, the loss on the rectifier is calcu-
lated based on the forward voltage of SiC Schottky Diodes. 27765 VOLUME 8, 2020 V.-B. Vu et al.: Improved LCL-L Compensation Topology for Capacitive Power Transfer in Electric Vehicle Charging current is at small value of 3A (Fig. 8a). Therefore, switching
losses are considered insignificant with the proposed system. It is noted that there are no crossing capacitances in hor-
izontal structure in [21] and therefore the associated losses
with the coupler are reduced. Future works can be con-
ducted by optimizing capacitive coupler to reduce the cross-
ing capacitances effects and improve the system efficiency. g
(3) Power losses on resonant network: g
(3) Power losses on resonant network: (a) On inductor L1p, where RL1p is the total AC and DC
resistances of L1p. (a) On inductor L1p, where RL1p is the total AC and DC
resistances of L1p. Ploss_L1p = I2
inRL1p
(A3) (A3) APPENDIX
REFERENCES EQUATIONS DERIVATION OF LOSS
CONTRIBUTIONS This section investigates the power losses of every single
component in the proposed converter. These calculations are
mainly based on [29]. Prec = 4
√
2
π VfwDIo + 2rDI2
o
(A7) (A7) (5) Losses on Aluminum capacitive coupler: it is difficult to
determine exact loss on the coupler PAlu. However, we can
estimate PAlu after determining other component’s losses. (A6) is the equation to determine PAlu. (1) The power conduction losses of the diode (PconD) and
MOSFET (PconMOS): Pcond = PconD + PconMOS
= 2
√
2
π VfwSIin
h
1 −sin
ϕ
2
i
+ 1
π RdsI2
in (π + ϕ + sin ϕ)
(A1) PAlu = Ptotal −
Pcond + Pswit + PlossL1p + PlossL1p
+ PlossC1p + PlossL1p + Prec
(A8) (A8) (A1) REFERENCES 33, no. 2, pp. 964–969, Feb. 2018. [13] L. J. Zou, Q. Zhu, C. Neste, and A. P. Hu, ‘‘Modelling single-wire
capacitive power transfer system with strong coupling to ground,’’ IEEE
J. Emerg. Sel. Topics Power Electron., to be published, doi: 10.1109/
jestpe.2019.2942034. VAN-BINH VU (Student Member, IEEE) received
the bachelor’s degree (talented program) from
the Hanoi University of Science and Technology,
Hanoi, Vietnam, in 2014, and the master’s degree
from Soongsil University, Seoul, South Korea,
in 2016, both in electrical engineering. He is cur-
rently pursuing the Ph.D. degree in power elec-
tronics with the School of Engineering, Newcastle
University, Newcastle upon Tyne, U.K. His main
research area includes the fields of power elec-
tronics, especially on wireless power transfer for electric vehicles. He has
authored or coauthored of four papers published in the top-tier IEEE journals
and has filled one patent on wireless power transfer technologies. He has
served as a regular reviewer for both the IEEE and IET journals. He has been
awarded a travel grant from the IEEE Industrial Electronics Society to attend
the 26th IEEE International Symposium on Industrial Electronics. [14] J. Dai and D. C. Ludois, ‘‘Single active switch power electronics for
kilowatt scale capacitive power transfer,’’ IEEE J. Emerg. Sel. Topics Power
Electron., vol. 3, no. 1, pp. 315–323, Mar. 2015. [15] J. Dai and D. C. Ludois, ‘‘Capacitive power transfer through a conformal
bumper for electric vehicle charging,’’ IEEE J. Emerg. Sel. Topics Power
Electron., vol. 4, no. 3, pp. 1015–1025, Sep. 2016. [16] S. Li, Z. Liu, H. Zhao, L. Zhu, C. Shuai, and Z. Chen, ‘‘Wireless
power transfer by electric field resonance and its application in dynamic
charging,’’ IEEE Trans. Ind. Electron., vol. 63, no. 10, pp. 6602–6612,
Oct. 2016. [17] S. Sinha, B. Regensburger, A. Kumar, and K. Afridi, ‘‘A very-high-power-
transfer-density GaN-based capacitive wireless power transfer system,’’ in
Proc. IEEE 5th Workshop Wide Bandgap Power Devices Appl. (WiPDA),
Oct. 2017, pp. 360–365. [18] S. Sinha, A. Kumar, B. Regensburger, and K. K. Afridi, ‘‘A new design
approach to mitigating the effect of parasitics in capacitive wireless power
transfer systems for electric vehicle charging,’’ IEEE Trans. Transp. Elec-
trific., vol. 5, no. 4, pp. 1040–1059, Dec. 2019. [19] B. Regensburger, S. Sinha, A. Kumar, J. Vance, Z. Popovic, and
K. K. Afridi, ‘‘Kilowatt-scale large air-gap multi-modular capacitive wire-
less power transfer system for electric vehicle charging,’’ in Proc. REFERENCES In which VfwS is the diode forward voltage, Rds is the
on-state resistance while ϕ and Iin are the conduction phase
angle and output RMS current of the Inverter. In this work, ϕ
is set as 175 degrees. [1] J. Dai and D. C. Ludois, ‘‘A survey of wireless power transfer and a critical
comparison of inductive and capacitive coupling for small gap applica-
tions,’’ IEEE Trans. Power Electron., vol. 30, no. 11, pp. 6017–6029,
Nov. 2015. [1] J. Dai and D. C. Ludois, ‘‘A survey of wireless power transfer and a critical
comparison of inductive and capacitive coupling for small gap applica-
tions,’’ IEEE Trans. Power Electron., vol. 30, no. 11, pp. 6017–6029,
Nov. 2015. [2] C. Liu, A. P. Hu, G. A. Covic, and N.-K.-C. Nair, ‘‘Comparative study of
CCPT systems with two different inductor tuning positions,’’ IEEE Trans. Power Electron., vol. 27, no. 1, pp. 294–306, Jan. 2012. (2) The switching losses on MOSFET: (2) The switching losses on MOSFET: The switching losses can be represented in (A2): tON and
tOFF are the turn-on and turn-off times of MOSFET, ID is the
drain current of each switch while VDS = Vin. [3] D. C. Ludois, M. J. Erickson, and J. K. Reed, ‘‘Aerodynamic fluid bearings
for translational and rotating capacitors in noncontact capacitive power
transfer systems,’’ IEEE Trans. Ind. Appl., vol. 50, no. 2, pp. 1025–1033,
Mar. 2014. [4] T. M. Mostafa, D. Bui, A. Muharam, R. Hattori, and A. P. Hu, ‘‘A capacitive
power transfer system with a CL network for improved system perfor-
mance,’’ in Proc. IEEE Wireless Power Transf. Conf. (WPTC), Jun. 2018,
pp. 1–5. Pswit = 4V DSIDfSW (tON + tOFF)
(A2) (A2) As analysis before, zero voltage switching at turn-on is
achieved for the proposed converter (i.e. tON = 0), while the
system is suffered from hard-switching at turn off. However,
we can see the turn-off loss is not significant as the turn-off [5] Y.-G. Su, S.-Y. Xie, A. P. Hu, C.-S. Tang, W. Zhou, and L. Huang,
‘‘Capacitive power transfer system with a mixed-resonant topology for
constant-current multiple-pickup applications,’’ IEEE Trans. Power Elec-
tron., vol. 32, no. 11, pp. 8778–8786, Nov. 2017. 27766 27766 VOLUME 8, 2020 V.-B. Vu et al.: Improved LCL-L Compensation Topology for Capacitive Power Transfer in Electric Vehicle Charging [6] L. Huang, A. P. Hu, A. K. Swain, and Y. REFERENCES Su, ‘‘Z-impedance compensation
for wireless power transfer based on electric field,’’ IEEE Trans. Power
Electron., vol. 31, no. 11, pp. 7556–7563, Nov. 2016. [26] Y. A. Liu, ‘‘High efficiency optimization of LLC resonant converter for
wide load range,’’ M.S. thesis, Dept. Electr. Comput. Eng., Virginia Poly-
tech. Inst. State Univ., Blacksburg, VA, USA, 2007. [27] J. Huh, S. W. Lee, W. Y. Lee, G. H. Cho, and C. T. Rim, ‘‘Narrow-
width inductive power transfer system for online electrical vehicles,’’ IEEE
Trans. Power Electron., vol. 26, no. 12, pp. 3666–3679, Dec. 2011. [7] X. Wu, Y. Su, X. Hou, X. Qing, and X. Dai, ‘‘Study on load adaptation
of capacitive power transfer system with a four-plate compact capacitive
coupler,’’ Electr. Eng., vol. 101, no. 3, pp. 733–742, Sep. 2019. [8] Y.-G. Su, Y.-M. Zhao, A. P. Hu, Z.-H. Wang, C.-S. Tang, and Y. Sun,
‘‘An F-type compensated capacitive power transfer system allowing for
sudden change of pickup,’’ IEEE J. Emerg. Sel. Topics Power Electron.,
vol. 7, no. 2, pp. 1084–1093, Jun. 2019. [28] V.-B. Vu, D.-H. Tran, and W. Choi, ‘‘Implementation of the constant
current and constant voltage charge of inductive power transfer systems
with the double-sided LCC compensation topology for electric vehicle
battery charge applications,’’ IEEE Trans. Power Electron., vol. 33, no. 9,
pp. 7398–7410, Sep. 2018. [9] T. M. Mostafa, A. Muharam, A. Patrick Hu, and R. Hattori, ‘‘Improved
CPT system with less voltage stress and sensitivity using a step-down
transformer on receiving side,’’ IET Power Electron., vol. 12, no. 10,
pp. 2634–2641, Aug. 2019. [29] B. X. Nguyen, D. M. Vilathgamuwa, G. H. B. Foo, P. Wang, A. Ong,
U. K. Madawala, and T. D. Nguyen, ‘‘An efficiency optimization scheme
for bidirectional inductive power transfer systems,’’ IEEE Trans. Power
Electron., vol. 30, no. 11, pp. 6310–6319, Nov. 2015. [10] J. Lian and X. Qu, ‘‘Design of a double-sided LC compensated capac-
itive power transfer system with capacitor voltage stress optimization,’’
IEEE Trans. Circuits Syst. II, Exp. Briefs, to be published, doi: 10.1109/
tcsii.2019.2918648. [11] R. H. Liang, C. K. Lee, and S. Hui, ‘‘Design, analysis and experimental
verification of a ball-joint structure with constant coupling for capacitive
wireless power transfer,’’ IEEE J. Emerg. Sel. Topics Power Electron., to
be published, doi: 10.1109/jestpe.2019.2938226. [12] F. Lu, H. Zhang, and C. Mi, ‘‘A two-plate capacitive wireless power transfer
system for electric vehicle charging applications,’’ IEEE Trans. Power
Electron., vol. REFERENCES Vu et al.: Improved LCL-L Compensation Topology for Capacitive Power Transfer in Electric Vehicle Charging VOLKER PICKERT (Member, IEEE) received the
Dipl. Ing. degree in electrical and electronic engi-
neering from RWTH Aachen, in 1994, and the
Ph.D. degree in power electronics from Newcastle
University, Newcastle upon Tyne, U.K., in 1997. He studied at the RWTH Aachen, Germany, and
Cambridge University, U.K. From 1998 to 1999,
he was an Application Engineer with Semikron
GmbH, Nuremberg, Germany. From 1999 to 2003,
he was the Group Leader at Volkswagen AG,
Wolfsburg, Germany, where he was responsible for the development of
electric drives for electric vehicles. In 2003, he was appointed as a Senior
Lecturer with the Electrical Power Group, Newcastle University, where he
became a Full Professor of power electronics, in 2011. In 2012, he became
the Head of the Electrical Power Group. He has published more than
150 book chapters, journal, and conference papers in the area of power
electronics and electric drives. His current research interests include power
electronics for transport applications, thermal management, health monitor-
ing techniques, and advanced nonlinear control. He received the IMarEST
Denny Medal for the Best Article in the Journal of Marine Engineering,
in 2011. In 2018, he received the Best Paper Award at the IEEE International
Conference on Computing Electronics and Communications Engineering
(iCCECE), Essex, U.K. He is regularly invited as a keynote speaker and
advises various governments on energy and transport related issues. In 2019,
he also became the Director of the UK’s EPSRC Doctoral Training Centre
in Power Electronics for Sustainable Electric Propulsion. He is the active
Editor-in-Chief of the IET Power Electronics journal. VAN-TUNG
PHAN
(Senior Member, IEEE)
received the bachelor’s degree in electrical and
electronic engineering from the Ho Chi Minh City
University of Technology, Ho Chi Minh City, Viet-
nam, and the Ph.D. degree in electrical engineer-
ing from the University of Ulsan, Ulsan, South
Korea. He was a Research Fellow with the Energy
Research Institute@NTU, Nanyang Technologi-
cal University (NTU), Singapore. He is currently
a Lecturer of electrical power engineering with
Newcastle University (Singapore Campus), Singapore. He has authored or
coauthored several internationally refereed journal articles, book chapters,
and conference papers on power electronics. REFERENCES IEEE
Appl. Power Electron. Conf. Expo. (APEC), Mar. 2018, pp. 666–671. Appl. Power Electron. Conf. Expo. (APEC), Mar. 2018, pp. 66 [20] A. Kumar, S. Pervaiz, C.-K. Chang, S. Korhummel, Z. Popovic, and
K. K. Afridi, ‘‘Investigation of power transfer density enhancement in
large air-gap capacitive wireless power transfer systems,’’ in Proc. IEEE
Wireless Power Transf. Conf. (WPTC), May 2015, pp. 1–4. [21] F. Lu, H. Zhang, H. Hofmann, and C. Mi, ‘‘A double-sided LCLC-
compensated capacitive power transfer system for electric vehicle charg-
ing,’’ IEEE Trans. Power Electron., vol. 30, no. 11, pp. 6011–6014,
Nov. 2015. MOHAMED DAHIDAH (Senior Member, IEEE)
received the Ph.D. degree in electrical engineering
from Multimedia University, Malaysia, in 2008. He was an Assistant Professor with the Depart-
ment of Electrical and Electronic Engineering, The
University of Nottingham, Malaysia Campus, until
November 2012. He is currently a Senior Lecturer
with the School of Engineering, Newcastle Univer-
sity, Newcastle Upon Tyne, U.K. He has authored
or coauthored a number of refereed journal and
conference papers. His research interests include modular multilevel con-
verters, SHE-PWM modulation technique for power electronics converters,
battery charger for EVs, solid state transformers, and renewable energy
integration. He is the Deputy Editor-in-Chief for the IET Power Electronics
and has been a regular reviewer for both the IEEE and IET journals. [22] H. Zhang, F. Lu, H. Hofmann, W. Liu, and C. Mi, ‘‘A four-plate compact
capacitive coupler design and LCL-compensated topology for capacitive
power transfer in electric vehicle charging application,’’ IEEE Trans. Power Electron., vol. 31, no. 12, pp. 8541–8551, Dec. 2016. [23] F. Lu, H. Zhang, H. Hofmann, and C. C. Mi, ‘‘A double-sided LC-
compensation circuit for loosely coupled capacitive power transfer,’’ IEEE
Trans. Power Electron., vol. 33, no. 2, pp. 1633–1643, Feb. 2018. [24] S. Sinha, B. Regensburger, K. Doubleday, A. Kumar, S. Pervaiz, and
K. K. Afridi, ‘‘High-power-transfer-density capacitive wireless power
transfer system for electric vehicle charging,’’ in Proc. IEEE Energy Con-
vers. Congr. Expo. (ECCE), Cincinnati, OH, USA, Oct. 2017, pp. 967–974. [25] V.-B. Vu, L. B. M. Kamal, J. Tay, V. Pickert, M. Dahidah, T. Logenthiran,
and V.-T. Phan, ‘‘A multi-output capacitive charger for electric vehicles,’’
in Proc. IEEE 26th Int. Symp. Ind. Electron. (ISIE), Edinburgh, U.K.,
Jun. 2017, pp. 565–569. 27767 VOLUME 8, 2020 VOLUME 8, 2020 V.-B. REFERENCES His current research interests
include on wireless power transfer systems for electric vehicles, dual active
bridge dc–dc converters, bi-directional ac–dc converters, dc–ac inverters, and
dc–dc converters in electrified vehicles and renewable energy applications,
such as solar and wind, on-board and off-board chargers, dc fast chargers
for electric vehicles; vehicle-to-grid interface technologies and grid-tied
photovoltaic micro-inverters and solar-powered electric vehicles charging
stations. He is a Senior Member of the IEEE Industrial Electronics Society,
the IEEE Power Electronics Society, the IEEE Vehicular Technology Society,
and the IEEE Industry Applications Society. He was a recipient of the
2013 IET Electric Power Applications Premium Award. 27768 VOLUME 8, 2020
|
https://openalex.org/W1964127298
|
https://www.scielo.br/j/jbchs/a/D9DSjGVZ4jFgRPwPypFQVyN/?lang=en&format=pdf
|
English
| null |
Traditional uses, chemistry and pharmacology of Croton species (Euphorbiaceae)
|
Journal of the Brazilian Chemical Society
| 2,007
|
cc-by
| 16,815
|
Traditional uses, Chemistry and Pharmacology of Croton species (Euphorbiaceae)
Antonio Salatino * Maria L Faria Salatino and Giuseppina Negri Departamento de Botânica, Instituto de Biociências, Universidade de São Paulo, CP 11461,
05422-970, São Paulo – SP, Brazil Croton é um gênero de Euphorbiaceae com cerca de 1300 espécies, amplamente distribuídas em
regiões tropicais do Novo e Velho Mundos. Várias espécies há muito tempo desempenham papel
importante nos usos tradicionais de plantas medicinais na África, Ásia e América do Sul. Tais usos
incluem tratamento de câncer, constipação intestinal, diarréia e outros problemas digestivos, diabetes,
feridas externas, febre, hipercolesterolemia, hipertensão, inflamação, vermes intestinais, malária,
dor, úlceras e obesidade. Várias espécies de Croton têm um látex vermelho que contém, em algumas
espécies, pró-antocianidinas e/ou alcalóides. Os alcalóides de Croton são a taspina ou substâncias
relacionadas a benzilisoquinolinas. Os diterpenos são muito comuns em Croton, correspondendo a
clerodanos, cembranóides, halimanos, cauranos, labdanos, ésteres do forbol, traquilobanos e
sarcopetalanos. Algumas espécies são aromáticas, devido à presença de óleos voláteis. Representantes
de novas classes de substâncias (fenilbutanóides, alcalóides glutarimidínicos e diterpenos
sarcopetalanos) têm sido isoladas de espécies de Croton. Enquanto laticíferos têm sido descritos em
espécies de Croton, até o momento não há estudos anatômicos sobre estruturas secretoras de óleos
voláteis. Parece haver afinidades químicas no gênero relacionadas à geografia, agrupando espécies
com (i) cauranos e/ou labdanos, (ii) traquilobanos e (iii) alcalóides. Os ensaios farmacológicos
freqüentemente vêm corroborando os usos tradicionais de espécies de Croton. Grande parte dos
ensaios farmacológicos trata do clerodano trans-desidrocrotonina, envolvendo uma grande diversidade
de efeitos, incluindo hipolipidêmico, hipoglicêmico, antiestrogênico e anticâncer. Efeitos citotóxicos
também vêm sendo observados em ensaios com alcalóides (taspina) e com diterpenóides secocaurenos,
labdanos e cembranóides. Vários outros efeitos de substâncias de Croton têm sido relatados, incluindo
anti-hipertensivos, antiinflamatórios, antimaláricos, antimicrobianos, antiespasmódicos,
antiulcerogênicos, antivirais e mio-relaxantes. Croton is a genus of Euphorbiaceae comprising around 1,300 species, widespread in tropical
regions of the Old and New Worlds. Several species have a long role in the traditional use of medicinal
plants in Africa, Asia and South America. Popular uses include treatment of cancer, constipation,
diabetes, digestive problems, dysentery, external wounds, fever, hypercholesterolemia, hypertension,
inflammation, intestinal worms, malaria, pain, ulcers and weight-loss. Several species of Croton
have a red sap, in some species containing proanthocyanidins and/or alkaloids. The latter may be
taspine or some of several benzylisoquinoline-like compounds. Diterpenes are very common in
Croton, corresponding to clerodanes, cembranoid, halimanes, kauranes, labdanes, phorbol esters,
trachylobanes and sarcopetalanes. Some species are aromatic due to the possession of volatile oils. J. Braz. Chem. Soc., Vol. 18, No. 1, 11-33, 2007.
Printed in Brazil - ©2007 Sociedade Brasileira de Química
0103 - 5053 $6.00+0.00 J. Braz. Chem. Soc., Vol. 18, No. 1, 11-33, 2007. Printed in Brazil - ©2007 Sociedade Brasileira de Química
0103 - 5053 $6.00+0.00 Review *e-mail: asalatin@ib.usp.br 1. Introduction used in traditional medicine to treat various diseases, being
reported to be balsamic, digestive, hypotensive, narcotic,
stomachic and tonic, and used to treat bronchitis, diarrhea
and dysentery.13 It has been used also against fever and
malaria, and as a substitute of Cinchona and Cascara.13 The World Health Organization estimates that around
80% of the world population in developing countries relies
on traditional plant medicines for primary healthcare
needs, of which a major proportion corresponds to plant
extracts or their active principles.1 Several species of Croton, such as C. lechleri, C. palanostigma, C. draconoides and C. urucurana, contain a
red viscous sap that is obtained upon making cuts on the
bark.14 The mature trees of C. lechleri L. produce a blood
red to yellowish-orange sap, which is highly prized for an
assumed ability to speed up the healing of wounds. On
account of its blood-like appearance, the latex is called in
Spanish either “sangre de drago” or “sangre de grado”,
translated into English as “dragon’s blood” and “blood of
the tree”, respectively.5 C. lechleri is among several species
of Croton used as purgative and tonic, and has a long history
of medicinal use among people of the South American
rainforest. It is a small-sized, fast growing tree, widely
distributed in the upper Amazonian valleys of Ecuador and
Peru. Its bark, when slashed, releases the blood-red sap,
highly astringent and used to treat wounds, as a haemostatic,
to accelerate healing and to protect lesions from infection. It is marketed as a health product.15-17 Many Euphorbiaceae are well known in different parts
of the world as toxic and/or medicinal. The high diversity of
the described effects is a reflex of the high chemical diversity
of this plant group. Croton is a large genus of Euphorbiaceae,
comprising around 1,300 species of trees, shrubs and herbs
distributed in tropical and subtropical regions of both
hemispheres. In the state of Rio de Janeiro alone 39 species
have been identified and some have been used in popular
medicine for many applications, including cancer.2 The genus
is rich in constituents with biological activities, chiefly
diterpenoids such as phorbol esters, clerodane, labdane,
kaurane, trachylobane, pimarane etc.3 Croton is also rich in
active alkaloids.4,5 Several species of the genus are aromatic,
indicating the presence of volatile oil constituents.6,7 As most
Euphorbiaceae, Croton species may contain latex, which is
red-colored in some species, a characteristic usually
associated with medicinal properties.8,9 C. Traditional uses, Chemistry and Pharmacology of Croton species (Euphorbiaceae)
Antonio Salatino * Maria L Faria Salatino and Giuseppina Negri Representatives of new classes of compounds (phenylbutanoids, glutarimide alkaloids, sarcopetalane
diterpenes) have been isolated from Croton species. While laticifers have been described in Croton
species, so far there are no anatomical studies about secretory structures of volatile oil. Few studies
about flavonoids have been carried out with Croton species. Chemical affinities are apparent in the
genus, grouping species with (i) kauranes and/or labdanes, (ii) trachylobanes and (iii) alkaloids. Pharmacological assays have frequently corroborated the traditional uses of Croton species. A great
part of pharmacological assays with Croton substances dealt with the clerodane trans-dehydrocrotonin,
a wide diversity of effects having been noticed, including hypolipidemic, hypoglycaemic, anti-
oestrogen and anti-cancer. Cytotoxic effects also have been observed in assays with alkaloids (taspine)
and with secokaurene, labdane and cembranoid diterpenes. Several other effects of Croton substances
have been registered, including anti-hypertensive, anti-inflammatory, antimalarial, antimicrobial,
antispasmodic, antiulcer, antiviral and myorelaxant. Keywords: Euphorbiaceae, Croton, diterpenes, alkaloids, flavonoids, volatile oils, antiulcer,
antitumoral, antimicrobial *e-mail: asalatin@ib.usp.br Traditional uses, Chemistry and Pharmacology of Croton species J. Braz. Chem. Soc. 12 1. Introduction malambo Karst., a small tree that grows in western
Venezuela and northern Colombia, is known as palomatias
and torco. An infusion of palomatias bark is used in traditional
medicine for treatment of diabetes, diarrhea, rheumatism,
gastric ulcer and as anti-inflammatory and analgesic.18 This paper gives an account of the species used in
traditional medicine, their chemistry and the pharmacological
activities of crude extracts and pure compounds. The data
collected are based on papers published up to March 2006. The data bases assessed include Biological and Chemical
Abstracts, and ISI Web of Science. C. nepetaefolius Baill., popularly called “marmeleiro-
vermelho”, is an aromatic plant native to northeastern Brazil,
used as a stomachic, carminative and for the treatment of
intestinal colic.19 Infusions or decoctions of the bark and
leaves are commonly used for their antispasmodic properties
and to relieve flatulence and to increase appetite.20 2.2. North America and Central America C. arboreous Millsp. is native in Mexico, where it is
known as “cascarillo”. In Tabasco and Chiapas a beverage
is prepared from the aerial parts and used as an auxiliary
anti-inflammatory in respiratory ailments.30 C. tiglium L. is an Asian shrub or tree (up to 12 m
tall), used in Chinese medicine as laxative.46 Other
traditional uses include tumors and cancerous sores
(www.hort.purdue.edu/newcrop/duke). The seeds are
source of a commercially available oil (croton oil), used
as purgative. C. californicus Müll. Arg. is a pale olive-green
perennial herb growing usually in sandy areas of the
Mojave Desert.31 Indians used to make a hot poultice from
its powdered leaves as pain reliever for rheumatism.32 C. tonkinensis Gagnep., popularly called “Kho sam
Bac Bo”, is a shrub native to northern Vietnam. The dried
leaves are prescribed for stomach-ache and have been used
in Vietnamese traditional medicine to treat burns,
abscesses, impetigo, dyspepsia and gastric/duodenal
ulcers. They are a component of recipes used to treat
urticaria, leprosy and psoriasis.47 C. draco Cham. & Schltdl. is a shrub native in Mexico
and Central America. It is one of the “sangre-de-drago”
plants, bearing a red sap widely used in traditional medicine,
including internal use for cough, flu, diarrhea and stomach
ulcers, and topically as wound healing for cuts, open sores,
herpes, anti-septic after tooth extraction and for oral sores.33 2.4. Asia 2.4. Asia 2.4. Asia C. kongensis Gagnep. is popularly known in Thailand
as “Plao Ngeon” or “Plao Noi”. It is frequently used in
folk medicine for dysmenorrhoea.38 2.1. South America C. palanostigma Klotzsch is one of the most widely
used medicinal plants of the South American tropics. The
red latex from the bark is used as wound-healer,21 treating
gastric ulcer and intestinal inflammation.22 Croton cajucara Benth., popularly known as
“sacaca”, grows in the Brazilian Amazonian region. It
is a traditional medicinal plant in the region, its leaves
and stem bark being used in the form of tea or pills for
the treatment of diabetes, high blood cholesterol levels
and gastrointestinal disturbances,10 as well as hepatic
disturbances and weight loss.11 C. schiedeanus Schlecht. is a tree native in Colombia,
being particularly widespread in the Amazonian and
Cundinamarca regions, where it is known as Almizclillo. This species is among the medicinal plants used in
Colombian traditional medicine for the treatment of
hypertension.23,24 C. celtidifolius Baill., commonly known as “sangue-
de-adave”, is a tree found in the Atlantic forest of
southern Brazil, mainly in Santa Catarina. Its bark and
leaf infusions have been popularly used for the
treatment of inflammatory diseases, leukemia, ulcer and
rheumatism.12 C. urucurana Baill. is a tree growing in Brazil from
Bahia to Rio Grande do Sul and Mato Grosso States. It is
one of the Croton species with bark containing red latex. C. urucurana is employed in traditional medicine due to
its analgesic and anti-inflammatory effects.25 It is used
also to treat wound infection, to accelerate wound-healing
and to treat rheumatism, cancer26 and other illnesses.27 C. eluteria Bennett., commonly known as “cascarilla”,
is a tropical shrub or small tree in the West Indies and
northern South America. Its bitter bark has been widely Vol. 18, No. 1, 2007 Salatino et al. 13 in association with C. sublyratus to treat gastric ulcers
and gastric cancers.42 C. zehntneri Pax. et Hoffm. is an aromatic plant native
in northeastern Brazil, where it is popularly called “canela-
de-cunhã” and used in traditional medicine mainly as
sedative, appetite stimulating, antianorexigen and for the
relief of gastrointestinal disturbances; its bark and leaves
are used as sweeteners in foods and drinks.28,29 Tribal people in India used various parts of C. roxburghii NP Balakr. against snake poisoning and to treat
infertility, fever and wounds.43 C. sublyratus Kurz is a tropical plant growing
extensively in southeastern Asian countries. In Thailand,
the plant is a popular medicine used as anti-helminthic44
and to treat dermatological problems.45 2.3. Africa Croton chemistry is considerably diverse. Terpenoids
are the predominant secondary metabolite constituents in
the genus, chiefly diterpenoids, which may belong to the
cembranoid, clerodane, neoclerodane, halimane,
isopimarane, kaurane, secokaurane, labdane, phorbol and
trachylobane skeletal types. Triterpenoids, either
pentacyclic or steroidal, have frequently been reported
for Croton species. Volatile oils containing mono and
sesquiterpenoids, and sometimes also shikimate-derived
compounds are not rare in the genus. Several species have
been reported as sources of different classes of alkaloids,
a fact that enhances considerably the importance of the
genus from the medicinal point of view. Phenolic
substances have frequently been reported, among which
flavonoids, lignoids and proanthocyanidins predominate. C. macrostachys Hochst. ex Rich. is a medium-sized
deciduous tree of East Africa. Its roots are used in Tanzania
as antidiabetic34 and the seeds are widely used in Somalia
as purgative.35 C. zambesicus Müll. Arg. (syn. C. amabilis Müll. Arg.;
C. gratissimus Burch.) is native in tropical west and central
Africa and used to treat fever, dysentery and convulsions.36
The leaf decoction is used in Benin as anti-hypertensive,
anti-microbial (urinary infections) and to treat malaria-
linked fever.3,37 C. zambesicus Müll. Arg. (syn. C. amabilis Müll. Arg.;
C. gratissimus Burch.) is native in tropical west and central
Africa and used to treat fever, dysentery and convulsions.36 The leaf decoction is used in Benin as anti-hypertensive,
anti-microbial (urinary infections) and to treat malaria-
linked fever.3,37 3.1. Diterpenes C. oblongifolius Roxb., popularly known as “chucka”,
is a middle-sized tree and grows in India and Thailand. The leaves are used as tonic, the flowers against flat
worms, the fruits to treat dysmenorrhoea, the seeds as
purgative,39 the bark to treat dyspepsia and the roots to
treat dysentery.40 The bark is also used to treat chronic
enlargement of the liver and remittent fever. It is applied
externally to the hepatic region in chronic hepatitis.41 In
Thai traditional medicine C. oblongifolius has been used Clerodane diterpenes, an extremely diverse group
of terpenoids with more than 800 known compounds,48
seem to be one of the prevalent classes of terpenoids in
Croton. The furane clerodanes with a lactone ring trans-
crotonin and trans-dehydrocrotonin (1) have been
isolated from the stem bark of C. cajucara, which
yielded also the nor-clerodanes cajucarin A, cajucarin
B, cajucarin-β, cajucarinolide and sacacarin.49,50 trans- Traditional uses, Chemistry and Pharmacology of Croton species 14 J. Braz. Chem. Soc. Crotonin and trans-dehydrocrotonin (t-DCTN) were
obtained from the aerial parts of C. cajucara.51 Other
sources of furano clerodanes are the stem barks of C. eluteria and C. urucurana. The former yielded
cascallin, cascarillone, cascarillins A-D,13 cascarillins
E-I, cascarilladione, eluterin K and pseudoeluterin B.52
Fattorusso et al.53 isolated ten new clerodanes (eluterins
A-J) from C.eluteria. The stem bark of C. urucurana
yielded sonderianin,27 15,16-epoxy-3,13(16)-clero-
datriene-2-one and 12-epi-methyl-barbascoate.25
Clerodanes were obtained from the bark of C. lechleri:
crolechinol and crolechinic acid,16 and the lactone
clerodanes korberin A and korberin B.17 Crolechinic
acid occurs as constituent from both the bark and red
sap of C. lechleri.54 Methyl barbascoate is a trans-
clerodane found as major diterpene in leaves of C. californicus.55 From shoots of C. schiedeanus, Puebla
et al.56 and Guerrero et al.57 isolated cis- and trans-
dehydrocrotonin and the new neo-clerodanes 5β-
hydroxy-cis-dehydrocrotonin and (12R)-12-hydroxy-
cascarillone. The acid fraction of shoot extracts of C. schiedeanus yielded two new cis-clerodanes (-)-methyl-
16-hydroxy-19-nor-2-oxo-cis-cleroda-3,13-dien-15,16-
olide-20-oate and (+)-15-methoxyfloridolide A.58
Palmeira et al.59 isolated the new clerodanes croto-
brasilins A and B from leaves and stems of C. brasiliensis (Spreng.) Müll. Arg. The labdane
crotonadiol (2) was obtained from the stem bark of C. zambesicus.60 From the same source, the clerodanes
crotocorylifuran and crotozambefuran A-C were
isolated, together with the trachylobanes 7β-acetoxy-
trachyloban-18-oic acid and trachyloban-7β,18-diol36
(3). Two clerodanes 3α,4β-dihydroxy-15,16-epoxy-12-
oxocleroda-13(16),14-dien-9-al and 3α,4β-dihydroxy-
15,16-epoxy-12-oxocleroda-13(16)14-diene were
isolated from bark of the Madagascarian C. 3.1. Diterpenes sublyratus contain the acyclic diterpene
alcohol plaunotol (5). Leaves of C. sublyratus are the
main source of this compound,45 although it may be
found also in the leaf chloroplasts of C. stellatopilosus
Ohba.67 C. oblongifolius has been a prolific source of
diterpenoids, including: (i) the clerodane 11-dehydro
(-) hardwickiic acid;68 (ii) the labdanes labda-
7,12(E),14-triene, labda-7,12(E),14-triene-17-al, labda-
7,12(E),14-triene-17-ol, labda-7,12(E),14-triene-17-oic
acid;69 (iii) the cembranoid diterpenes crotocembranoic
acid and neocrotocembranal70 (6); (iv) the cytotoxic
labdane diterpenoids 2-acetoxy-3-hydroxy-labda-
8(17),12(E)-14-triene, 3-acetoxy-2-hydroxy-labda-
8(17),12(E)-14-triene, and 2,3-dihydroxy-labda-
8(17),12(E),14-triene were also isolated from the stem
bark of C. oblongifolius;71 (v) the labdane nidorellol,
the furoclerodane croblongifolin and the clerodane
crovatin;72 (vi) the kaurane (-)ent-kaur-16-en-19-oic
acid;40 (vii) the halimanes crotohalimaneic acid (7),
crotohalimoneic acid and 12-benzoyloxycroto-
halimaneic acid;73 (viii) new labdane-type diterpenoids
3-oxygenated ent-manoyl oxide derivatives with a 8,13-
epoxytricyclic ring system and hydroxylabdandienes.74
(-)-Hardwickiic acid (8) was isolated from C. californicus,75 C. draco33 and C. aromaticus L., a
species with red latex native in Sri Lanka.76 Draconine,
a clerodane diterpene, was obtained from C. draco
(Rodriguez-Hahn and co-workers, apud Hernandez and
Delgado77). C. draco contains also 9(11)-dehydro-
kaurenoic acid, (5R*,8R*,9S*,10R*)-15,16-epoxy-12-
o x o c l e r o d a - 3 , 1 3 ( 1 6 ) , 1 4 - t r i e n e - 1 8 - a c i d ,
(2S*,5R*,8R*,9R*,10S*,12S*)-12,20;15,16-diepoxy-
2-hydroxy-20-oxocleroda-3,13(16),14-triene-18-acid
and the new clerodane (5R*,8R*,9S*,10R*)-15,16- Crotonin and trans-dehydrocrotonin (t-DCTN) were
obtained from the aerial parts of C. cajucara.51 Other
sources of furano clerodanes are the stem barks of C. eluteria and C. urucurana. The former yielded
cascallin, cascarillone, cascarillins A-D,13 cascarillins
E-I, cascarilladione, eluterin K and pseudoeluterin B.52
Fattorusso et al.53 isolated ten new clerodanes (eluterins
A-J) from C.eluteria. The stem bark of C. urucurana
yielded sonderianin,27 15,16-epoxy-3,13(16)-clero-
datriene-2-one and 12-epi-methyl-barbascoate.25
Clerodanes were obtained from the bark of C. lechleri:
crolechinol and crolechinic acid,16 and the lactone
clerodanes korberin A and korberin B.17 Crolechinic
acid occurs as constituent from both the bark and red
sap of C. lechleri.54 Methyl barbascoate is a trans-
clerodane found as major diterpene in leaves of C. californicus.55 From shoots of C. schiedeanus, Puebla
et al.56 and Guerrero et al.57 isolated cis- and trans-
dehydrocrotonin and the new neo-clerodanes 5β-
hydroxy-cis-dehydrocrotonin and (12R)-12-hydroxy-
cascarillone. The acid fraction of shoot extracts of C. 3.1. Diterpenes hovarum
Leandri.61 From leaves of plants of the same species,
the clerodanes 3,12-dioxo-15,16-epoxy-cleroda-
13(16),14-dien-9-al and 3α,4β-dihydroxy-15,16-epoxy-
19-nor-12-oxo-cleroda-5(10),13(16),14-triene were
isolated.62 From leaves of C. zambesicus the trachy-
lobane ent-trachyloban-3β-ol was obtained.37 From the
same source ent-18-hydroxy-trachyloban-3-one and the
isopimarane-type diterpenoid isopimara-7,15-dien-3β-
ol were also obtained.3 C. tonkinensis, a species native
in Vietnam, has been a prolific source of ent-kaurane-
type diterpenoids. From leaves of this species ent-7β- ent-18-acetoxy-7α-hydroxykaur-16-en-5-one, plus the
new compounds ent-1β-acetoxy-7α,14β-dihydroxy-
kaur-16-en-15-one and ent-18-acetoxy-7α,14β-
dihydroxykaur-16-en-15-one were isolated,47 as well as
obtained four new ent-kauranes: ent-1α,14α-diacetoxy-
7β-hydroxykaur-16-en-15-one, ent-1α,7β-diacetoxy-
14α-hydroxykaur-16-en-15-one, ent-18-acetoxy-14α-
hydroxykaur-16-en-15-one and ent-(16S)-18-acetoxy-
7β-hydroxykauran-15-one.65 Recently, Giang et al.66
obtained again from leaves of C. tonkinensis the six
new ent-kauranes ent-7β-acetoxy-11-α-hydroxykaur-
16-en-15-one, ent-18-acetoxy-11-α-hydroxykaur-16-
en-15-one, ent-11-α-hydroxykaur-16-en-18-oic acid,
ent-15-α-18-dihydroxykaur-16-ene, ent-α-18-dia-
cetoxy-7-β-hydroxykaur-16-en-15-one and ent-(16S)-
1-α-14-α-diacetoxy-7-β-hydroxy-17-methoxykauran-
15-one. Besides clerodane and kaurane derivatives, the
leaves of C. sublyratus contain the acyclic diterpene
alcohol plaunotol (5). Leaves of C. sublyratus are the
main source of this compound,45 although it may be
found also in the leaf chloroplasts of C. stellatopilosus
Ohba.67 C. oblongifolius has been a prolific source of
diterpenoids, including: (i) the clerodane 11-dehydro
(-) hardwickiic acid;68 (ii) the labdanes labda-
7,12(E),14-triene, labda-7,12(E),14-triene-17-al, labda-
7,12(E),14-triene-17-ol, labda-7,12(E),14-triene-17-oic
acid;69 (iii) the cembranoid diterpenes crotocembranoic
acid and neocrotocembranal70 (6); (iv) the cytotoxic
labdane diterpenoids 2-acetoxy-3-hydroxy-labda-
8(17),12(E)-14-triene, 3-acetoxy-2-hydroxy-labda-
8(17),12(E)-14-triene, and 2,3-dihydroxy-labda-
8(17),12(E),14-triene were also isolated from the stem
bark of C. oblongifolius;71 (v) the labdane nidorellol,
the furoclerodane croblongifolin and the clerodane
crovatin;72 (vi) the kaurane (-)ent-kaur-16-en-19-oic
acid;40 (vii) the halimanes crotohalimaneic acid (7),
crotohalimoneic acid and 12-benzoyloxycroto-
halimaneic acid;73 (viii) new labdane-type diterpenoids
3-oxygenated ent-manoyl oxide derivatives with a 8,13-
epoxytricyclic ring system and hydroxylabdandienes.74
(-)-Hardwickiic acid (8) was isolated from C. californicus,75 C. draco33 and C. aromaticus L., a
species with red latex native in Sri Lanka.76 Draconine,
a clerodane diterpene, was obtained from C. draco
(Rodriguez-Hahn and co-workers, apud Hernandez and
Delgado77). C. draco contains also 9(11)-dehydro-
kaurenoic acid, (5R*,8R*,9S*,10R*)-15,16-epoxy-12-
l
d
3 1 3 ( 1 6 ) 1 4 t i
1 8
i d ent-18-acetoxy-7α-hydroxykaur-16-en-5-one, plus the
new compounds ent-1β-acetoxy-7α,14β-dihydroxy-
kaur-16-en-15-one and ent-18-acetoxy-7α,14β-
dihydroxykaur-16-en-15-one were isolated,47 as well as
obtained four new ent-kauranes: ent-1α,14α-diacetoxy-
7β-hydroxykaur-16-en-15-one, ent-1α,7β-diacetoxy-
14α-hydroxykaur-16-en-15-one, ent-18-acetoxy-14α-
hydroxykaur-16-en-15-one and ent-(16S)-18-acetoxy-
7β-hydroxykauran-15-one.65 Recently, Giang et al.66
obtained again from leaves of C. tonkinensis the six
new ent-kauranes ent-7β-acetoxy-11-α-hydroxykaur-
16-en-15-one, ent-18-acetoxy-11-α-hydroxykaur-16-
en-15-one, ent-11-α-hydroxykaur-16-en-18-oic acid,
ent-15-α-18-dihydroxykaur-16-ene, ent-α-18-dia-
cetoxy-7-β-hydroxykaur-16-en-15-one and ent-(16S)-
1-α-14-α-diacetoxy-7-β-hydroxy-17-methoxykauran-
15-one. Besides clerodane and kaurane derivatives, the
leaves of C. 3.1. Diterpenes schiedeanus yielded two new cis-clerodanes (-)-methyl-
16-hydroxy-19-nor-2-oxo-cis-cleroda-3,13-dien-15,16-
olide-20-oate and (+)-15-methoxyfloridolide A.58
Palmeira et al.59 isolated the new clerodanes croto-
brasilins A and B from leaves and stems of C. brasiliensis (Spreng.) Müll. Arg. The labdane
crotonadiol (2) was obtained from the stem bark of C. zambesicus.60 From the same source, the clerodanes
crotocorylifuran and crotozambefuran A-C were
isolated, together with the trachylobanes 7β-acetoxy-
trachyloban-18-oic acid and trachyloban-7β,18-diol36
(3). Two clerodanes 3α,4β-dihydroxy-15,16-epoxy-12-
oxocleroda-13(16),14-dien-9-al and 3α,4β-dihydroxy-
15,16-epoxy-12-oxocleroda-13(16)14-diene were
isolated from bark of the Madagascarian C. hovarum
Leandri.61 From leaves of plants of the same species,
the clerodanes 3,12-dioxo-15,16-epoxy-cleroda-
13(16),14-dien-9-al and 3α,4β-dihydroxy-15,16-epoxy-
19-nor-12-oxo-cleroda-5(10),13(16),14-triene were
isolated.62 From leaves of C. zambesicus the trachy-
lobane ent-trachyloban-3β-ol was obtained.37 From the
same source ent-18-hydroxy-trachyloban-3-one and the
isopimarane-type diterpenoid isopimara-7,15-dien-3β-
ol were also obtained.3 C. tonkinensis, a species native
in Vietnam, has been a prolific source of ent-kaurane-
type diterpenoids. From leaves of this species ent-7β-
hydroxy-15-oxokaur-16-en-18-yl acetate63 and ent-1α-
acetoxy-7β,14α-dihydroxy-kaur-16-en-15-one64 (4)
were isolated. From the same source the known ent-
kauranes ent-7α,14β-dihydroxykaur-16-en-15-one and Salatino et al. 15 Vol. 18, No. 1, 2007 A and B, and crotinsulactone.82 From shoots of C. salutaris Casar, a tree from southeast Brazil, Itokawa
et al.83 obtained the acyclic diterpenes [(10E)-3,12-
dihydroxy-3,7,11,15-tetramethyl-1,10,14-hexa-
decatrien-5,13-dione, (6E,10E)-3,12-dihydroxy-
3,7,11,15-tetramethyl-1,6,10,14-hexadecatetraen-5,13-
dione and (6Z,10E)-3,12-dihydroxy-3,7,11,15-tetra-
methyl-1,6,10,14-hexadecatetraen-5,13-dione] and a
tricyclic diterpene [12-hydroxy-13-methylpodocarpa-
9,11,13-trien-3-one], in addition to sonderianol. A
prenylbisabolone diterpene with insecticidal effect was
obtained from the Jamaican C. linearis Jacq.84 In
addition to yucalexins B-6 and P-4, roots of C. sarcopetalus Muell., a shrub native in Bolivia and
central and northwestern Argentina, contain diterpenes
bearing the novel skeleton sarcopetalane: sarcopetaloic
acid and two sarcopetalolides, namely sarcopetal-15-
en-3-one-12,13-olide (10) and sarcopetal-15-en-3β-ol-
12,13-olide; the same plant source contains junceic acid
and stress metabolites.85,86 epoxy-12-oxocleroda-3,13(16),14-triene-18-acid.33
Secokauranes have been obtained from leaves of C. kongensis.38 Trachylobanes, such as 3α-19-dihydroxy-
trachylobane, 3α-18,19-trihydroxytrachylobane,
trachyloban-18-oic acid and trachylobane-19-oic acid,
were obtained from roots of C. macrostachys, together
with neoclerodan-5,10-en-19,6β,20,12-diolide.34
Trachyloban-19-ol and methyl trachyloban-19-oate
were isolated from the stem bark of C. robustus.78 From
C. zambesicus, Baccelli et al.79 isolated the novel
diterpenes ent-18-hydroxytrachyloban-3β-ol and ent-
18-hydroxyisopimara-7,15-diene-3β-ol. The pimarane
diterpene 3β-hydroxy-19-O-acetyl-pimara-8(9),15-
dien-7-one (9) and the labdane 2α,3α-dihydroxy-labda-
8(17),12(13),14(15)-triene were isolated from leaves
of the Thailandese C. joufra Roxb.80 Two novel
diterpenes ent-trachyloban-3-one and crotinsularin were
isolated from aerial parts of the Australian C. insularis
Baillon.81 From the same plant source, three new
clerodanes were obtained, namely furocrotinsulolides Traditional uses, Chemistry and Pharmacology of Croton species J. 3.1. Diterpenes Braz. Chem. Soc. 16 Phorbol esters are produced by species of
Euphorbiaceae and Thymelaeaceae.87 Among Croton
species, only C. tiglium, native and cultivated in India,
has been extensively studied as source of phorbol
derivatives, having been shown to contain tigliane phorbol
esters. The main irritant component of C. tiglium seeds is
12-O-tetradecanoylphorbol-13-acetate, a tumor promoter
used in experimental mice cancer research.88 Other phorbol
esters of C. tiglium seeds include 13-O-acetylphorbol-20-
linoleate, 13-O-tigloylphorbol-20-linoleate, 12-O-
acetylphorbol-13-tigliate, 12-O-decanoylphorbol-13-(2-
methylbutyrate), 12-O-tigloylphorbol-13-(2-methyl-
butirate) and 12-O-acetylphorbol-13-decanoate89 (11), 12-
O-tetradecanoylphorbol-13-acetate and 12-O-(2-
methylbutiroyl)-phorbol-13-dodecanoate. 9 0
C. macrostachys also probably contains phorbols in its
laxative seeds.35 Small amounts of a phorbol ester was
detected in flowers of C. draco.33 methylisoeugenol as main constituent.86 The volatile oil
of the aerial parts of the Venezuelan C. malambo is
composed mainly of methyleugenol, methylisoeugenol,
elemicine, isoelemicine veratral and δ-cadinol.94 The
volatile oil of the shoots of C. cuneatus Miq., another
Venezuelan species, comprise α-11-eudesmene, methyl-
eugenol, methylisoeugenol, 4-α-seleniol, cedryl-propyl
ether, τ-cadinol, cubenol, veratral and agarospirol.94 The
same plant source yielded the sesquiterpene sellin-11-en-
4α-ol. From the second category belongs the volatile oil
of C. cajucara, which is believed to contain sesqui-
terpenes,95 although it is known to be rich in the
monoterpene linalool.96-98 Linalool and cineol are
monoterpenoids seemingly relatively frequent in
Croton. Both are relevant constituents of the volatile
oil of the African C. oligandrum Pierre ex Hutch.99
Linalool is among the major constituents of the volatile
oil of C. stellulifer Hutch., an endemic species of S. Tomé and Príncipe; this oil contains kessane, a
sesquiterpenoid oxide not found elsewhere in Croton.100 3.2. Volatile oils and their constituents The volatile oil of leaves of C. sacaquinha Benth. also
contains linalool, in addition to the sesquiterpenes
β-elemene and germacrene D.7 1,8-Cineol is an
important constituent of the oil of C. sonderianus Müll. Arg.101 Murillo et al.33 obtained the nor-terpenoids
9-dehydrovomifoliol and 2,3-dihydrovomifoliol from
seeds and 1-hydroxyjunenol from the bark of C. draco. The sesquiterpenes τ-cadinol and epicubenol and the
acetophenone xanthoxylin were found in shoots of C. hieronymi Griseb.102 The sesquiterpenes caryophyllene
oxide and trans-caryophyllene were found to
predominate in leaves, while caryophyllene oxide and
cubenol predominate in stems of C. sellowii Baill., a
species from northeast Brazil. In the roots of the same
species, mesitylene and the sesquiterpenes α, β and
γ-eudesmol were the main compounds.103 Shoots of
C. 3.1. Diterpenes arboreus Millsp., an arboreous species from Mexico,
contain the sesquiterpenes 5α,7α,10βH-3-patchoulen-
2-one, 5α,7α,10βH-4-(14)- patchoulen-2α-ol and
9α,10β-dihydroxy-2β,4β-peroxy-1α,5β,7αH-guaiane,
along with 14 other known compounds, including the
clerodane junceic acid.30 Another species with volatile
oil containing predominantly sesquiterpenes (caryo-
phyllene, caryophyllene oxide, germacrenes, cadinol,
spathulenol) is the Venezuelan C. ovalifolius Vahl.104
Leaves of C. zambesicus have volatile oil containing
mostly sesquiterpenes (β-caryophyllene, caryophyllene
oxide and α-copaene), monoterpenes (linalool and
β-pinene) and smaller amounts of diterpenes
(trachylobanes, isopimaranes and kaurenes).105 3.3. Pentacyclic triterpenoids and steroids morphinandienones and tetrahydroprotoberberine
alkaloids. Glutarimide alkaloids and a new class of
sesquiterpene guaiane-type alkaloids have recently been
obtained from Croton species. The aerial parts of C. draco contain β-sitosterol,
stigmasterol and the new sterol ergasterol-5α-8α-
endoperoxide.77 From leaves of C. betulaster Müell. Arg.,
a species from northeast Brazil, Barbosa et al.106 obtained
the triterpenes lupenone, lupeol, 3-oxo-22-hydroxy-
hopane, 3-oxo-20-β-hydroxytaraxastane, 3-oxo-20-
hydroxylupane, 3-oxo-olean-12-en-28-oic acid, 3-oxo-
olean-18-en-28-oic acid, and 3-oxo-cycloart-24E-en-26-
oic acid. The triterpene acetyl aleuritolic acid (12) has
been obtained from two Croton species native in Brazil,
C. cajucara49,50 and C. urucurana.27,107 From methanol
extracts of the latter species, stigmasterol, β-sitosterol,
campesterol and β-sitosterol-O-glucoside were obtained. Shoots of C. hieronymi contain a C-25 analog of trans-
phytol and triterpenes, such as α- and β-amyrin, lupeol
and hop-22(29)-en-3β-ol, in addition to the squalene
derivatives all-trans-10-methylene-2,6,10,14,18,22-
hexaen-3-ol and 2,6,15,19,23-pentamethyltetracosa-
2,6,10(28),14,22,28-hexaene-11-ol, and steroids, such as
cholesterol, cholest-8(14)-en-3β-ol, stigmasterol,
gramisterol, sitosterol, campesterol, 22-dihydrobrassi-
casterol, lophenol (13), isofucosterol, stigmasterol,
cholest-4-en-3-one, ergosta-4-22-dien-3-one and
sitostenone.102 Taspine (14), an unusual alkaloid with a dilactone
structure resembling elagic acid and one nitrogen atom
not included in a heterocyclic ring, was found in the red
latex of three species, C. draco,33,108 C. lechleri9,109 and C. palanostigma.110 Taspine has been obtained also from plant
sources of benzylisoquinolines and biogenetically related
alkaloids, such as Berberidaceae and Magnoliaceae. From
leaves of C. lechleri other alkaloids, probably related
biogenetically to taspine, have also been isolated, such as
glaucine (15), isoboldine, magnoflorine, norisoboldine,
thaliporphine5 and sinoacutine.111 Tetrahydroproto-
berberine alkaloids have been reported for C. hemiargyreus
Müll. Arg. and C. flavens L. From leaves and stems of the
former Amaral and Barnes4 isolated 2,10-dihydroxy-3,10-
dimethoxy-8β-methyldibenzo[a,g]-quinolizidine
(hemiargyrine) (16), in addition to glaucine, oxoglaucine,
salutaridine (17) and norsalutaridine. The tetrahydro-
protoberberine alkaloids scoulerine and coreximine and
the morphinanedienone alkaloids salutaridine and
salutarine, in addition to sebiferine, norsinoacutine and
flavinantine, were isolated from specimens from Barbados
of C. flavens by Eisenreich et al.112 From leaves of C. flavens, Charris et al.113 isolated (-)-amuronine. From
shoots of C. salutaris Barnes and Soeiro114 isolated
salutarine and salutaridine, the latter a biosynthetically
precursor of morphine.115 Isoboldine and laudanine were
found in the ethanolic extracts of leaves and twigs from
C. celtidifolius.116 Stuart and Graham117 verified that C. linearis synthesizes crotonosine through linearisine. Murillo et al.33 obtained two known aporphines from the
bark and wood of C. draco. The β-carboline alkaloids 2-
ethoxycarbonyltetrahydroharman and 6-hydroxy-2-
methyltetradydroharman (18) were obtained from plants
of C. 3.3. Pentacyclic triterpenoids and steroids moritibensis Bail., a species from northeastern
Brazil.118 The aerial parts of C. cuneatus yielded the new
glutarimide alkaloids julocrotol, isojulocrotol and
julocrotone (19), in addition to julocrotonine.119 Anabasine
and the novel guaiane-type alkaloids muscicapines A, B
and C (20) were isolated from roots of the Northeastern
Brazilian C. muscicapa Müll. Arg.120 Murillo et al.33 obtained two known aporphines from the
bark and wood of C. draco. The β-carboline alkaloids 2-
ethoxycarbonyltetrahydroharman and 6-hydroxy-2-
methyltetradydroharman (18) were obtained from plants
of C. moritibensis Bail., a species from northeastern
Brazil.118 The aerial parts of C. cuneatus yielded the new
glutarimide alkaloids julocrotol, isojulocrotol and
julocrotone (19), in addition to julocrotonine.119 Anabasine
and the novel guaiane-type alkaloids muscicapines A, B
and C (20) were isolated from roots of the Northeastern
Brazilian C. muscicapa Müll. Arg.120 3.2. Volatile oils and their constituents 3.2. Volatile oils and their constituents Although most Euphorbiaceae are plants not known
as aromatic, some Croton species contain volatile oils. Other species have not been reported as bearing volatile
oils, although they were shown to possess sesquiterpenes
commonly found in volatile oils. The volatile oils of
several species contain phenylpropanoids and terpenoids
(mono and sesquiterpenes), while from other species only
terpenoids have been obtained. The volatile oils of leaves and stem bark of C. nepetaefolius belong to the first above category. It contains
monoterpenes (such as 1,8-cineole) and terpineol,
sesquiterpenes (such as bicyclogermacrene), and volatile
phenylpropanoids (such as methyleugenol) and the
acetophenone xanthoxylin.91 The volatile oil from shoots
of C. zehntneri seems to be composed mainly of
phenylpropanoids, such as anethole, methyleugenol and
estragole.6,92 Methyleugenol is also found with terpenoids
in volatile oils of other Croton species, such as the Central
American C. jimenezii Standl. & Valerio.93 A volatile oil
was obtained from the roots of C. sarcopetalus, with trans- Vol. 18, No. 1, 2007 Salatino et al. 17 3.5. Proanthocyanidins Alkaloids are not common in Euphorbiaceae, but some
Croton species are notable for their alkaloids. The most
frequent Croton alkaloids are compounds identical or
similar to substances found in Ranunculales, i.e. alkaloids
biogenetically related to benzylisoquinolines, such as Tannins are polyphenols virtually ubiquitous in plants. They are medicinally important if occurring in high
proportions in the plants. They may be formed by the
combination of catechin monomers (the so-called Traditional uses, Chemistry and Pharmacology of Croton species
J. Braz. Chem. So Traditional uses, Chemistry and Pharmacology of Croton species 18 J. Braz. Chem. Soc. (4-α→8)-gallocatechin and gallocatechin-(4-α→8)-
gallocatechin-(4-α→8)-epigallocatechin. Similar tannic
compounds were obtained from the red latex of C. panamensis (Klotzsch) Müll. Arg.126 Dalbo et al.127
detected in a fraction of the crude extract of C. celtidifolius
bark polymeric and dimeric proanthocyanidins, the latter
represented mainly by catechin-(4-α→8)-catechin and
gallocatechin-(4-α→8)-catechin. proanthocyanidins), or by ester bounded units of glucose,
gallic and/or ellagic acid (hydrolysable tannins). So far,
only proanthocyanidins have been characterized in Croton
species. Proanthocyanidins have been reported as important
active principles of sangre de drago species, i.e. species
containing red latex.121 In such species, proanthocyanidins
with several degrees of polymerisation and molecular size
comprise almost 90% of the red latex dry weight. C. urucurana also contains proanthocyanidins and catechins
in its red latex.122 A specific proanthocyanidin coded SP-
303, with average molecular weight of 2,100 daltons,123
was isolated from the red latex of C. lechleri.124 Tannins
seem to be important active principles also of C. zambesicus.125 Cai et al.15 detected proanthocyanidins as
major constituents, accounting for up to 90% of the dry
weight of C. lechleri red latex. Monomers such as (+)-
catechin, (-)-epicatechin, (+)-gallocatechin, (-)-
epigallocatechin and dimeric procyanidins B-1 and B-4
were isolated. Dimers and trimers were also obtained and
characterized as catechin-(4-α→8)-epigallocatechin,
gallocatechin-(4-α→8)-epicatechin, gallocatechin-(4-
α→6)-epigallocatechin, catechin-(4-α→8)-gallocatechin- 3.7. Other classes of compounds methoxyflavones artemetin (22), casticin, chrysosplenol-
D and penduletin. The n-hexane extract of the Caribbean
and Central American C. ciliatoglanduliferus Ort. yielded the highly methoxylated flavonols retusin and
pachypodol.128 Flavonoid data is still too scarce to
establish a pattern for Croton. The available data suggest
a common presence of flavonols and flavones as free
highly methoxylated aglycones. It is important to note,
however, that no investigation directed specifically to
flavonoids, which is currently made in hydro-alcoholic
extracts,129 has been carried out with Croton species. Hence, it is quite possible that flavonol and/or flavone
glycosides (present in green tissues of most angiosperm
species), have not been detected in the so far reported
Croton chemical analyses. methoxyflavones artemetin (22), casticin, chrysosplenol-
D and penduletin. The n-hexane extract of the Caribbean
and Central American C. ciliatoglanduliferus Ort. yielded the highly methoxylated flavonols retusin and
pachypodol.128 Flavonoid data is still too scarce to
establish a pattern for Croton. The available data suggest
a common presence of flavonols and flavones as free
highly methoxylated aglycones. It is important to note,
however, that no investigation directed specifically to
flavonoids, which is currently made in hydro-alcoholic
extracts,129 has been carried out with Croton species. Hence, it is quite possible that flavonol and/or flavone
glycosides (present in green tissues of most angiosperm
species), have not been detected in the so far reported
Croton chemical analyses. Scopoletin (a coumarin) was obtained from the wood
extract of C. draco; the seeds of the same species contain
p-hydroxybenzaldehyde and p-methoxybenzoic acid, while
the dried sap yielded 3,4,5-trimethoxycinnamic alcohol.33
Phenylbutanoids, an interesting class of compounds known
to occur in some genera of angiosperms, were obtained
from shoots of C. schiedeanus by Puebla et al.132 These
authors isolated (2S)-7,9-dimethoxyrhododendrol (25),
(2S)-acetoxy-7,9-dimethoxyrhododendrol and (2S)-2,8-
diacetoxy-7,9-dimethoxyrhododendrol. The formation of
this class of phenolics has been proposed to occur via
decarboxylative condensation of 4-coumaroil-CoA with
malonyl-CoA to produce C6C4
skeletons.133 The novel
compounds 4-(2-hydroxyethyl)-benzoic acid and 2,5-
dihydroxy-phenylethanol were isolated from the red sap of
C. panamensis.126 Lichexanthone was obtained from the
aerial parts of C. cuneatus.119 From the same source,
Hernandez and Delgado77 obtained a mixture of polyprenols,
castaprenol-11 being the major compound. Using high-
temperature high resolution gas chromatography coupled
to mass spectrometry (HT-HRGC-MS), Pereira et al.2,134
characterized in shoots of C. hemiargyreus more than 100
compounds, including alkaloids, amino acids, terpenoids
and wax esters. 3.6. Flavonoids and other phenolic substances Flavonols and/or flavones are ubiquitous in vascular
plants, at least in green tissues. From the red latex of C. draco108 and C. panamensis126 myricitrin (myricetin-3-
O-rhamnoside) was isolated. Leaves of C. cajucara
yielded kaempferol-3,7-dimethyl ether and 3,4’,7-
trimethyl ether,50 while shoots of C. schiedeanus contain
quercetin-3,7-dimethyl ether.24 From leaves of C. betulaster Müll. Arg., Barbosa et al.106 obtained 5-
hydroxy,7,4’-dimethoxyflavone, and from leaves of C. hovarum Leandri, Krebs and Ramiarantsoa62 isolated the
flavone C-glycoside vitexin (21). From leaves and stems
of C. brasiliensis, Palmeira et al.59 isolated the 3- Vol. 18, No. 1, 2007 19 Salatino et al. Salatino 3.7. Other classes of compounds Among the compounds detected is a high
molecular mass triterpenoid (simiarenol) and esters of
amyrine with fatty acids containing carbon chains above
20 atoms. The polyalcohols 1L-1-O-myo-inositol and neo-
inositol were isolated from shoots of C. celtidifolius.135
Benzoyl-methylpolyols were isolated from C. betulaster
and Croton luetzelburgii Pax & K. Hoffm.136 Furanoarabino-
galactan, a polysaccharide, is the main component found Only recently were phenyl propanoids reported for the
first time in Croton. From the aerial parts of C. hutchinsonianus Hos., a species native in Thailand, two
new compounds were isolated, namely 3’-(4”-hydroxy-
phenyl)-propyl benzoate and 3’-(4”-hydroxy-3”,5”-
dimethoxyphenyl)-propyl benzoate (23), together with the
known 3’-(4”-hydroxy-3”-methoxyphenyl)-propyl
benzoate.130 Lignoids are common in plant groups bearing
benzylisoquinolines and related alkaloids (derived
biosynthetically from tirosine), such as Ranunculales and
Magnoliales. As was commented above, some Croton
species possess this class of alkaloids. However, only one
lignoid has so far been found in Croton, the dihydro-
benzofuran lignan 3’,4-O-dimethylcedrusin (24). It is
interesting to note that this lignan co-occur with taspine,
having been found in C. lechleri9 and C. palanostigma,131
both species with red latex. Traditional uses, Chemistry and Pharmacology of Croton species 20 J. Braz. Chem. Soc. The most common class of compounds of Croton is
represented by diterpenoids. Apparently, clerodane is the
widest spread class of diterpenoids in Croton, which has
been found in species from America (e.g. C. cajucara),
Africa (e.g. C. macrostachys) and Asia (e.g. C. oblongifolius). Phorbol esters (such as 11) seem also to
be widespread in Croton, having been found in species
from Asia (e.g. C. tiglium), Africa (e.g. C. macrostachys)
and America (e.g. C. californicus). Thus, the common
possession of clerodanes and phorbol esters by groups
of species should be viewed as retention of an ancient
characteristic, rather than a mark of natural affinity. in the gum exsudate from C. urucurana.137 The peptide
derivatives aurentiamide acetate and N-benzoylpheny-
lalanine were isolated from shoots of C. hieronymi.102
Cyclopeptides were reported for the red latex of C. draco.108 Figure 1. Chemical and geographical affinities among Croton species. 4. Chemotaxonomy Tomé and Príncipe, Gulf of Guinea,
west of Africa) and C. zambesicus (Africa). Assuming the possession of diterpenes as an ancient
chemical feature in Croton, the apparent absence of
diterpenoids in extant Croton species indicates that the
ability to produce or accumulate such compounds was lost
in the evolutionary process. It has been claimed that
terpenoid production by plants is more costly than other
secondary metabolites.139 It seems that loss of diterpenoids
in Croton occurred during the spread of Croton species into
America. Benzylisoquinoline-like alkaloids (15-17) are an
important group of secondary metabolites found so far
exclusively in American Croton species, namely C. celtidifolius, C. draco, C. flavens, C. hemiargyreus, C. lechleri, C. linearis, C. palanostigma and C. salutaris
(Figure 1). Again, it is worth noting that C. lechleri and C. flavens emerge inside a same clade in Croton molecular
phylogeny.138 Alkaloids from other classes also occur in
American Croton species, such as glutarimide alkaloids in
C. cuneatus (Venezuela), β-carbolines in C. moritibensis
(Brazil) and anabasine and guaiane-type alkaloids in C. muscicapa (Brazil). There seems to be in Croton a nearly
mutual exclusiveness between diterpenes and alkaloids. Among the mentioned alkaloid-bearer species only C. salutaris has been reported to bear alkaloids and
diterpenoids. C. lechleri and C. palanostigma are akin also
due to the common possession of taspine (14), high amounts
of proanthocyanidins and the lignoid 3’,4-dimethylcedrusin
(24) (Figure 1). As commented above, lignans are common
in plant groups that produce benzylisoquinolines. In
addition, these two species, plus C. celtidifolius, have red
latex. Red sap occurs also in non-alkaloidal species, such
as C. cajucara and C. urucurana, which share the possession
of the rare triterpenoid acetyl aleuritolic acid (12). A molecular phylogeny of Croton, including plants
with known chemistry, would be extremely helpful to
clarify trends in the chemical evolution of the genus. 5.1. Antilipidemic Extracts of C. cajucara leaves showed significant
reductions in the serum total cholesterol, low-density
lipoprotein cholesterol and triglyceride levels, as well as
a significant elevation in the HDL/total cholesterol ratio
in treated rats compared with the control group.140
Experiments treating rats with water extracts11 gave support
to the popular use of C. cajucara bark in loss-weight
programs. Treated rats had lower weight gain than control
rats, the sensitivity of the lipolytic responses to
isoprenaline and adrenaline being significant higher in
adipocytes from treated rats. 4. Chemotaxonomy Instead, kauranes, labdanes and trachylobanes seem to
be more restricted in their distribution. C. draco, C. kongensis, C. sublyratus and C. tonkinensis are chemically
akin due to the possession of kauranes. Labdanes occur in
C. joufra, C. oblongifolius and C. zambesicus. Trachylobanes have been found in C. insularis, C. macrostachys, C. robustus and C. zambesicus. It is worth
noting that, with the exception of C. draco (a species with
a highly diverse chemistry), all other species with these
diterpene classes do not occur in the New World, most of
them occurring in Asia, C. zambesicus in Africa and C. Given the huge dimension of the genus and the so far
minute sampling of species chemically studied, no definite
conclusions can be drawn about chemical relationships
among Croton species. However, the present picture of
the distribution of compound classes is suggestive of
chemical patterns in the genus (Figure 1) and encourages
proposals of chemical affinities, which in turn may
stimulate further work on prospects of Croton biologically
active compounds. Figure 1. Chemical and geographical affinities among Croton species. Salatino et al. 21 Vol. 18, No. 1, 2007 insularis in Australia (Figure 1). A recent molecular
phylogeny of Croton has been published,138 but unfortunately
few chemically studied species were included. Nonetheless,
it is interesting that C. kongensis and C. zambesicus are
grouped in a same clade. Other classes of diterpenoids,
such as cembranoid, pimarane and isopimarane diterpenes
seem to be highly restricted. In Croton, cembranoids have
been found only in C. oblongifolius, which so far is the
sole known halimane diterpenoid bearer. In Croton,
isopimaranes are so far exclusive of C. zambesicus and
pimaranes of C. joufra (Figure 1). Up until now, C. sarcopetalus is the sole known source of sarcopetalane
diterpenoids. cuneatus, C. draco, C. hieronymi, C. jimenezii, C. malambo,
C. nepetaefolius, C. oligandrum, C. ovalifolius, C. sacaquinha, C. sarcopetalus, C. selowii, C. sonderianus, C. stellulifer, C. zambesicus and C. zentneri (Figure 1). Among
these species only C. arboreous, C. cajucara, C. draco, C. sarcopetalus and C. zambesicus have been reported to possess
diterpenoids and volatile oils. Volatile oils seem to be absent
from alkaloidal species; so far only C. cuneatus was shown
to bear both volatile oil and alkaloids, the latter represented
by the rare glutarimide-type alkaloids. Most aromatic species
occur in America, the exceptions being C. oligandrum
(Africa), C. stellulifer (S. 5.9. Antioxidant C. celtidifolius bark has anti-inflammatory and
antioxidant activity, which results from the direct action of
constituents on specific targets, such as cyclooxygenase.12
C. urucurana red latex has antioxidant effect against lipid
peroxidation and free radical scavenging activity.26 C. lechleri sap possesses significant antioxidant activity against
the oxidative damages induced by apomorphine and
hydrogen peroxide in Saccharomyces cerevisiae and maize
plantlets.152 Leaf extracts of C. cajucara were observed to
exert antioxidant effects against the free radical DPPH and
in paraquat treated yeast cells.154 5.10. Myorelaxant, antispasmodic and anti-hypertensive The volatile oil of the bark and leaves of C. nepetaefolius, which contains mainly 1,8-cineole,
methyleugenol and terpineol, exerted antispasmodic
effect on gastrointestinal tissues and antihypertensive
activity in the cardiovascular system.155 Intravenous
treatment with the volatile oil decreases mean aortic
pressure and heart rate in either anaesthetized or non-
anaesthetized rats.156 The myorelaxant activity is
coherent with the use of C. nepetaefolius in traditional
medicine as antispasmodic. Its large efficacy as a
myorelaxant agent associated with its low acute toxicity
makes it an agent of therapeutic potential.91 Aqueous
and ethanolic extracts of C. schiedeanus have a
decreasing effect on blood pressure, probably by means
of an antihypertensive rather than hypotensive
effect.23,157 The antihypertensive activity and vaso-
dilatory effects of C. schiedeanus are attributed to a 5.4. Immunomodulatory, antibacterial, antifungal and antiviral The red latex of C. lechleri showed antioxidant or pro-
oxidant properties and stimulated or inhibited the
phagocytosis. In addition, sangre de drago has immuno-
modulatory activity, inhibiting the proliferation of activated
T-cells.9 The latex is also known to have antioxidant, free
radical scavenging147 and antiviral activities.124 In addition,
it has revealed anti-bacterial properties.142 The red latex from
C. urucurana inhibited the growth of the fungi Tricophyton
tonsurans, Tricophyton mentagrophytes, Tricophyton rubrum,
Microsporum canis and Epidermophyton floccossum,
showing a potential utility as an alternative treatment for
dermatophytosis.148 Phytochemical analyses demonstrated in
the product the presence of catechins, such as gallocatechin
and epigallocatechin, substances previously known to exert
antifungal activity. The red latex of C. draco and its ethyl
acetate and ethyl ether extracts exhibited high inhibition on
the classical activation pathway of the complement system
using hemolytic assay.108 5.8. Mutagenic The red sap of C. lechleri showed mutagenic activity
for strain TA1535 of Salmonella typhimurium, in the
presence of metabolic activation, and weak mutagenic
activity for strain TA98; mutagenic activity was also
noted in the haploid Saccharomyces cerevisiae strain
XV185-14c.152 Methanolic extracts of C. cajucara bark
were tested in mice for mutagenicity and antimuta-
genicity.153 No statistical significant results were
obtained, indicating that, at the concentrations tested,
the extracts have no mutagenic effect. 5.5. Antimalarial Several Croton species are traditionally used as
antimalarials throughout endemic malarial areas. Antiplasmodial activity was demonstrated in vitro for
C. pseudopulchellus Pax., a species from southern
Africa.149 Shoot methanolic extracts from aerial parts of
C. lobatus L. (a widespread species in tropical America,
from Florida to Argentina) were active toward
Plasmodium falciparum 3D7 chloroquine sensitive
strains, while root methanolic extracts inhibited growth
of K1 resistant strains.150 5.2. Wound healing, gastric effects and protection Extracts of the bitter bark of cascarilla (C. eluteria) has
been shown to strengthen the histamine-stimulated gastric
acid secretion, giving experimental support to the use of
cascarilla in bitter preparations aimed to improve
digestion.141 The “sangre de grado” of C. lechleri has shown
wound-healing activity142 in cutaneous disorders and, orally,
in a dilute form to facilitate the healing of gastric ulcers,
reducing ulcer size and bacterial content of the ulcer.143 The volatile oil from the bark of C. cajucara has been
shown to exert gastric ulcer healing activity, as well as
protection of the gastric mucosa.144 Low molecular weight
sesquiterpenes appear to be important active constituents
of the volatile oil.95 Efficacy of the oil seems to be based
on its ability to stimulate local mucus synthesis and
prostaglandin production by the gastric mucosa.145 Another trend that possibly occurred in the chemical
evolution of Croton is the loss of diterpenoids and the
substitution by mono and sesquiterpenes, mainly as volatile
oil constituents, accompanied sometimes by shikimate
derived volatiles (e.g. methyl eugenol, and anethole). Volatile
oils have been reported from C. arboreous, C. cajucara, C. Traditional uses, Chemistry and Pharmacology of Croton species
J. Braz. Chem. Soc. Traditional uses, Chemistry and Pharmacology of Croton species Traditional uses, Chemistry and Pharmacology of Croton species 22 J. Braz. Chem. Soc. 5.3. Anti-diarrheic C. lechleri sap possess antimutagenic properties and
may inhibit the proliferation of human leukemic cells.151 The red sap from C. urucurana showed promising
potential for the control of pathologies associated with
secretory diarrhea.122 Proanthocyanidin SP–303 (from
sangre de drago of C. lechleri) is a potent inhibitor of cholera
toxin-induced fluid accumulation and chloride secretion.146 5.13. Reliever of insect bites swelling The red latex of C. lechleri relieves swelling of insect
bites.143 6.2. Tumor promotion Although
taspine is probably an important and active constituent of
the red latex, it cannot be considered the main responsible
for these activities, and other constituents, probably
proanthocyanidins, should be also involved.9 Orally
administered, the volatile oil of C. nepetaefolius promoted
a dose-dependent antinociceptive effect in hot-plate test.158 5.6. Anticancer Shoots of C. hieronymi have shown strong activity
against lung A-549 carcinoma cells and mouse lymphoma
and some activity against human colon carcinoma.102 The
dichloromethane extract of leaves of C. zambesicus
showed in vitro cytotoxicity against human cervix
carcinoma cells.37 The red latex of C. lechleri has been
shown to have anti-tumor activity.142 Salatino et al. 23 Vol. 18, No. 1, 2007 the Amazon region for the control of hyperlipidemy and
associated pathologies.50,163 Hypolipidemic effects were
observed by in assays with t-DCTN from C. cajucara.164,165
In addition to hypolipidemic action, t-DCTN exhibited
hypoglycemic effect in alloxan-induced diabetic, but not
in normal rats.140 the Amazon region for the control of hyperlipidemy and
associated pathologies.50,163 Hypolipidemic effects were
observed by in assays with t-DCTN from C. cajucara.164,165 synergistic activity among flavonoids and terpenoids.24
The volatile oil of C. zehntneri has relaxing effect on
smooth muscle, which supports the use of C. zehntneri
in traditional medicine as a gastrointestinal antispas-
modic, an activity that may in part be attributed to
estragole.28,29 In addition to hypolipidemic action, t-DCTN exhibited
hypoglycemic effect in alloxan-induced diabetic, but not
in normal rats.140 6.3. Anticancer 6.3. Anticancer t-DCTN (1) exhibits anti-tumor efficacy and
immunomodulatory actions in vivo, which may be related
to its chemical structure.176 DCTN and its synthetic
derivative dimethylamide-crotonin (DCR) inhibit HL60
cells growth in vitro partly by apoptosis induction and
cell differentiation, but do not cause serious damage to
immune cells.177,178 However, t-DCTN is not cytotoxic (and
also not genotoxic) to bone marrow cells of Swiss mice
submitted to acute intraperitoneal treatment in vivo179 and
antimutagenic with regard to cyclophosphamide, in
particular if administered by gavage.180 Correa et al.181
developed a β-cyclodextrin complex to improve delivery
of DCTN. A lower cytotoxicity of the complex
β-cyclodextrin-DCTN to V79 fibroblasts and rat cultured
hepatocytes, compared to free t-DCTN, suggests that such 5.13. Reliever of insect bites swelling 5.12. Central depressive The volatile oil from the bark and leaves of C. zehntneri
produced central depressive effects in rats without anxiety
alterations.162 6.2. Tumor promotion Phorbol esters (such as 11) are known co-carcinogenic
agents, being involved in the transformation of normal human
epithelial cells through infection by Epstein-Barr virus.166
The commercially available oil of Croton derives from the
Asiatic C. tiglium. This oil is the source of phorbol esters,
tetracyclic diterpenes isolated from the seeds, whose co-
carcinogenic properties have been the object of numerous
investigations. Although they have been consumed
medicinally for two millennia,167 these natural products are
the most powerful tumor promoters known.168-170 C. tiglium
is still utilized in homoeopathy and in combination with some
sort of acupuncture (as a constituent of Baunscheidt oil). Its
main irritant component, 12-O-tetradecanoylphorbol-13-
acetate (TPA) was used as a standard tumor promoter in mice
in experimental cancer research,88 but Castagna et al.171 and
Nishizuka172 noted that phorbol-12-myristate-13-acetate is
also effective. In addition, the latter compound is hardly
metabolized by the cell.173 Several phorbol esters were
prepared with polar functional groups terminating their C12
and/or C13 acyl chains. Such phorbol ester lipophilic domain
is thought to be responsible for redistributing a phorbol-
protein complex to the plasma membrane.174 Biotrans-
formation of phorbol esters by intestinal bacteria has been
successfully achieved.175 Crude leaf extracts of C. cajucara exhibited significant
antinociceptive effect in rats.10 The red latex of C. lechleri
revealed strong anti-inflammatory activity. Although
taspine is probably an important and active constituent of
the red latex, it cannot be considered the main responsible
for these activities, and other constituents, probably
proanthocyanidins, should be also involved.9 Orally
administered, the volatile oil of C. nepetaefolius promoted
a dose-dependent antinociceptive effect in hot-plate test.158
The aqueous extract of the aerial parts of C. cuneatus had
significant activity against plantar inflammation induced
by bovine serum albumin.134 The volatile oil of C. sonderianus had antinociceptive effect in tests with oral
administration, but was inactive in hot-plate tests.160 An
antinociceptive effect of the volatile oil of C. zehntneri
was evidenced, most likely associated with anti-
inflammatory activity.6 The aqueous extract of C. malambo
bark administered intraperitoneally showed antino-
ciceptive and anti-inflammatory effects, comparable to
acetylsalicylic acid and sodium diclofenac.161 C. celtidifolius bark has anti-inflammatory activity.12 Crude leaf extracts of C. cajucara exhibited significant
antinociceptive effect in rats.10 The red latex of C. lechleri
revealed strong anti-inflammatory activity. 6.4. Antibacterial, antifungal and antiviral 8,9-Secokauranes from C. kongensis exhibited anti-
mycobacterial activity at the minimum inhibitory
concentration of 6.25-25.0 μg mL-1.38 Plaunotol (5) has
exhibited activity against twenty methicillin-resistant and
fourteen methicillin-sensitive strains of Staphylococcus
aureus.187,188 The results suggested that plaunotol might
be useful in the prevention of infection and skin care for
patients with atopic dermatitis. Tannins are known to complex with proteins, acting
as multidentate ligands and thus being able to bind
simultaneously to more than one point to the protein
surface.189 Proanthocyanidin SP-303 has been extensively
studied and has shown activity against a variety of virus:
respiratory syncytial virus, parainfluenza type-1 and type-
3, and influenza A and B viruses;190 herpes virus;191 genital
and anogenital herpes virus, in treatment of patients with
AIDS;192 several DNA and RNA virus, including hepatitis
viruses.143 C. tiglium seeds contain anti-HIV-1 phorbol
esters, 12-O-acetylphorbol-13-decanoate (11) and 12-O-
decadienoylphorbol-13-(2-methylbutyrate) that inhibit the
cytopathic effect of HIV-1; 12-O-tetradecanoylphorbol-
13-acetate (TPA) is even more active than the mentioned
phorbol esters against HIV-1.89,90 Catechin and acetyl aleuritolic acids, compounds
obtained from C. urucurana, are effective against S. aureus
and Salmonella typhimurium; acetyl aleuritolic acid showed
minimum inhibitory concentration ten fold higher than
catechin.27 Alviano et al.98 observed that the volatile oil from
leaves of C. cajucara, composed mostly by linalool, inhibits
the growth of Candida albicans, Lactobacilus casei,
Porphyromonas gengivalis, S. aureus and Streptococcus
mutans, all involved in diseases of the oral cavity. Among
these microorganisms, the authors noted that linalool is
active almost exclusively against Candida albicans, and
that the volatile oil is not toxic to mammalian cells. Phorbol esters may exert potent co-carcinogenic
activity but, among them, many exert beneficial biological
effects without tumor promotion, such as prostratin.183
Actually, some naturally occurring phorbol esters are
tumor inhibitors.90 12-O-Tigloylphorbol-13-decanoate has
been shown to have activity against the P 388 lymphocytic
leukemia in mice.184 The phenylpropyl benzoates 3'-(4"-hydroxy-3",5"-
dimethoxyphenyl)-propyl benzoate, 3'-(4"-hydroxy-
phenyl)propyl benzoate and 3'-(4"-hydroxy-3"-methoxy-
phenyl)-propyl benzoate, obtained from stems of C. hutchinsonianus, were shown to exert effect against
Candida albicans.130 Plaunotol (5), an acyclic diterpene present in C. sublyratus
leaves, has recently been shown to have anti-cancer activity
through inhibition of angiogenesis.185 Anethole, a phenyl-
propanoid constituent of C. zehntneri volatile oil, has been
shown to have anti-carcinogenic effect.186 Derivatives of phorbol esters have been evaluated as
inhibitors of proliferation of HIV-1. 6. Pharmacological Effects of Pure Compounds 6.1. Anti-diabetic, anti-hyperlipidemic, anti-hyper-
cholesterolemic 6.1. Anti-diabetic, anti-hyperlipidemic, anti-hyper-
cholesterolemic Pharmacological studies carried out with the terpenoids
t-DCTN (1), trans-crotonin, acetyl aleuritolic acid (12)
and with plant extracts showed a striking correlation with
the traditional therapeutic use of C. cajucara species in Traditional uses, Chemistry and Pharmacology of Croton species J. Braz. Chem. Soc. 24 J. Braz. Chem. Soc. complex may be useful for in vivo dehydrocrotonin
administration. cells were obtained with the glutarimide alkaloids
julocrotol, isojulocrotol and julocrotone (19) isolated from
the aerial parts of C. cuneatus.119 The furoclerodane croblongifolin from C. oblongifolius
showed significant cytotoxicity against BT474, CHAGO,
HEP-G2, KATO3 and SW620 tumor cell lines.72 Among
Croton diterpenes, not only clerodanes have been found to
exert anticancer activity. Non-specific, moderate cytotoxicity
agains human breast ductal carcinoma (BT474), human
undifferentiated lung carcinoma (CHAGO), human liver
hepatoblastoma (HEP-G2), gastric carcinoma (KATO3) and
colon adenocarcinoma (SW620) tumor cell lines were shown
by the following labdanes from C. oblongifolius: labda-
7,12(E),14-triene-17-al; 17-hydroxy-labda-7,12(E),14-triene;
17-acetoxylabda-7,12(E),14-triene; 15-hydroxylabda-
7,13(E)-diene-17,12-olide; labda-7,13(E)-diene-17,12-olide
and 12,17-dihydroxylabda-7,13(E)-diene.39 The labdanes
from C. oblongifolius 2,3-dihydroxy-labda-8(17),12(E),14-
triene showed non-specific, moderate, cytotoxicities against
all the cell lines BT474, CHAGO, HEP-G2, KATO3 and
SW620, whereas 2-acetoxy-3-hydroxy-labda-8(17),12(E),14-
triene and 3-acetoxy-2-hydroxy-labda-8(17),12(E),14-triene
were less active, because their ability to form hydrogen bond
with certain receptors in tumor cells made them more
selective but less active.71 Also halimane and cembranoid
diterpenes from C. oblongifolius showed antitumoral activity,
but 12-benzoyloxycrotohalimaneic acid was inactive.73
Neocrotocembranal (6) inhibited platelet aggregation induced
by thrombin and exhibited cytotoxicity against P-338 cells
in vitro.70 ent-Kauranes of C. tonkinensis also have been
shown to be cytotoxic.66 Block et al.37 reported for the first
time cytotoxic activity of a trachylobane (ent-trachyloban-
3β-ol), a constituent of leaves of C. zambesicus. The same
compound has recently been shown to induce apoptosis in
human promyelocytic leukemia cells in a concentration-
dependent manner.182 6.10. Anti-oestrogenic t-DCTN (1) obtained from C. cajucara was tested for
anti-oestrogenic activity using immature rats for bioassay
of oestrogen and regularly cycling rats of proven fertility
for antiimplantation effect. The compound prevented
increases of uterine wet weights, but did not affect
blastocyst implantation.214 6.8. Vasorelaxant, hypotensive and bradycardiac 6.8. Vasorelaxant, hypotensive and bradycardiac t-DCTN (1) was shown to reduce the mean arterial
pressure and heart rate in a dose-dependent manner in
normotensive rats and to relax the tonic contraction in
isolated rat aortic rings induced by phenylephrine.212 The
neo-clerodanes (12R)-12-hydroxycascarillone, 5β-
hydroxy-cis-dehydrocrotonin, cis- and trans-dehydro-
crotonin from C. schiedeanus relaxed aort rings.57 A
cooperactivity between diterpenoids and quercetin-3,7-
dimethyl ether (ayanin, the predominant flavonoid in C. schiedeanus leaves) was observed in the relaxing effect. Flavonoids have been assigned a role as protectors against
vascular accidents. A vasorelaxant activity of quercetin-
3,7-dimethyl ether from C. schiedeanus was observed,
the activity probably being influenced by hydroxylation
at positions 3’ and 4’ of the B ring.24 Plaunotol is an anti-peptic ulcer agent,201 commercially
available under the name Kelnac.45 The anti-ulcer effect
of plaunotol is probably linked to its activity against
Helicobacter pilory.202,203 6.6. Antispasmodic and anti-diarrheic The volatile oils of some South-American Croton
species are antispasmodic. Cineole, methyleugenol and
terpineol, constituents of C. nepetaefolius volatile oil, have
been reported to have myorelaxant and antispasmodic
effects in laboratory animals.153 Experimental results
suggest that C. nepetaefolius volatile oil induces relaxation
of guinea-pig ileum. 204 Anethole and estragole, major
components of the volatile oil of C. zehntneri, are effective
relaxants of skeletal muscles.205 6.9. Central anti-depressive Fischer et al.147 described a method to optimize the
preparation of an extract from the bark latex of C. lechleri. The extract was named SB-300 and contains on average
70.6% by weight of the proanthocyanidin SP-303. The
authors suggest its use for the treatment of fluid loss in
acute watery diarrhea. SP-303 is the active agent of Provir,
a C. lechleri derived product with antisecretory
properties206,207 and hence useful to control fluid loss and
diarrhea. SP-303 has been indicated particularly for
patients of AIDS, common victims of diarrhea.208 While the volatile oil of C. zehntneri exhibited
depressive effects in models using rats (see above), methyl-
eugenol, its major constituent, was shown to have anti-
depressive effect in experiments based on the same
models.213 t-DCTN (1) showed strong antiulcerogenic activity.195,196 The A-ring of both crotonin and DCTN is not directly
involved in the antiulcerogenic activity.197 t-DCTN has good
antiulcerogenic activity; a long term use of which, however,
may induce liver damage.198 A semi-synthetic crotonin,
namely 4SRC, was synthesized and it was shown to have a
significant preventive effect against gastric ulcer induced
by different agents.199 Presence of a lactone moiety or
Michael acceptor is probably essential for the anti-
ulcerogenic effect, a mechanism of gastric cytoprotection
being mediated by an action on prostaglandin biosynthesis,
and by a Michael reaction between the SH-containing
compounds of the mucosa on the Michael acceptors present
in antiulcerogenic compounds.200 The volatile oil of C. zehntneri was shown to have
antinociceptive activity in mice,6 and the volatile oil of
C. cajucara has anti-inflammatory and antinociceptive
effects in rodents.209 Cajucarinolide, a diterpene from C. cajucara, was shown to possess anti-inflammatory activity
and to inhibit bee venom phospholipase A2 in vitro.210
DCTN, the main component of bark extracts of C. cajucara, has anti-inflammatory and analgesic effects.50,.211 A chromatographic analysis from the ethyl acetate
soluble fraction of the crude extract of the stem bark of
C. celtidifolius showed predominantly proanthocyanidins,
which exhibited antinociceptive effects in tests using
intraperitonial injections of chemical stimulants.127 When previously administered orally the volatile oil
from the bark of Croton cajucara significantly reduced
the gastric injury induced in rats.95,144 6.5. Wound-healing, gastric effects and protection active sesquiterpenes are 5α,10β-4(15)-eudesmen-1β,6β-
diol, spathulenol, 5α,10β-3-eudesmen-1β,6α-diol and the
diterpenoid junceic acid. t-DCTN (1) showed strong antiulcerogenic activity.195,196 6.4. Antibacterial, antifungal and antiviral Among them 12-O-
acetylphorbol-13-decanoate has been shown to be the most
potent.193,194 Taspine (14) is active against KB and V-79 cells, a
fact that makes it a likely responsible for the purported
anticancer activity of C. lechleri red sap.142 Positive results
in antitumor tests using breast carcinoma and hepatoma Salatino et al. Vol. 18, No. 1, 2007 25 6.5. Wound-healing, gastric effects and protection 7. Concluding Remarks: Points Deserving
Further Work A lot of further work is also necessary about traditional
uses of Croton species, not only in Brazil, but also in other
tropical regions of both New and Old Worlds. Indeed, the
number of Croton species reported in the literature as
sources of traditional medicines is too small, if it is
considered the high diversity of Croton species, chemistry
and pharmacology. Table 1 summarizes traditional uses and effects
observed in pharmacological assays with species of
Croton and respective substances. Taking into account
the magnitude of the genus, the list of species is still too
scant. However, a quick examination of the table contents
suffices to give an idea of the wide diversity of (i) health
problems, which people from many parts of the world
treat with the aid of Croton plant parts, (ii) classes of
Croton secondary metabolites, and (iii) pharmacological
effects experimentally observed, using crude extracts and
pure compounds. For these reasons, much chemical and
pharmacological work is needed about Croton. So far
only one species of Croton is known as source either of
halimane and cembranoid (C. zambesicus), or
isopimarane (C. zambesicus) or sarcopetalane (C. sarcopetalus) diterpenoids. Paul Berry estimates that
there are 354 known Croton species native in Brazil,
one of the important hotspots of Croton diversity
(www.cria.org.br/eventos/tdbi/flora/presentations/
PaulBe). Nevertheless, as yet Croton literature reports
results of chemical and/or pharmacological research
related with only the following Brazilian species: C. betulaster, C. brasiliensis, C. cajucara, C. celtidifolius,
C. eluteria, C. lobatus, C. luetzelburgii, C. moritibensis,
C. muscicapa, C. nepetaefolius, C. palanostigma, C. sacaquinha, C. salutaris, C. sellowii, C. sonderianus,
C. urucurana and C. zehntneri. Fortunately, a rapid
inventory in the ISI Web of Science reveal that For several species in Table 1, there is good
agreement between popular uses and experimentally
observed effects, in harmony with Fairnsworth’s
observation216 that popular uses of medicinal plants
frequently are coherent with the pharmacological
effects of the main active principles of the respective
plant sources. Part of the future pharmacological
investigations should center their focus on several
popular uses of Croton species, which so far have
received no scientific support, as are the cases of C. californicus (rheumatism pain), C. draco (several
traditional uses), C. eluteria (bronchitis, fever, malaria),
C. kongensis (dysmenorrhoea), C. macrostachys
(diabetes, purgative), C. malambo (diarrhea), C. oblongifolius (several traditional uses), C. palanostigma
(wounds, digestive problems), C. roxburghii (infertility,
fever, wounds), C. sublyratus (intestinal worms,
dermatological problems), C. 6.12. Anti-leishmanial and anti-malarial activity The linalool-rich volatile oil from leaves of C. cajucara
was shown to be a potent agent against Leishmania
amazonensis. The DL50 for promastigotes is 8.3 ng mL-1
and for amastigotes, 8.7 ng mL-1. The inhibitory
concentration for L. amazonensis promastigotes growth
is extremely low (MIC 85.0 pg mL-1), and the oil has no
cytotoxic effects against mammalian cells.97 8,9-
Secokauranes obtained from C. kongensis were shown to
have anti-malarial activity with IC50 1.0-2.8 μg mL-1.38 Some trends in Croton research should be rechecked. It is evident from the Reference list of the present work
that some species have been the object of relatively many
investigations, such as C. cajucara, C. lechleri, C. urucurana and C. zehntneri, whilst most Croton species
have so far been neglected by chemists and pharma-
cologists. 6.11. Insect growth inhibition and insecticidal approximately 30 papers have been published annually
from 2000 to 2005 about Croton chemistry and/or
pharmacology. It is important to note that figures much
higher would emerge if articles were also counted
without Croton in their titles, but dealing with chemical
or pharmacological aspects of the genus. Hence, Croton
species and their substances are among the highly studied
themes in chemistry of natural products, pharmacology
and ethnopharmacology, and new data are rapidly
accumulating. It is expected that many novelties will
rapidly enlarge the current knowledge about Croton
species, their substances and corresponding pharma-
cological effects. The diterpene fraction from C. linearis showed lethal
effect on insects.84 A prenylbisabolane diterpene from C. linearis has insecticidal effect.215 The same comment
applies to hardwickiic acid, a diterpene present in C. aromaticus and C. californicus.76 6.7. Anti-inflammatory and antinociceptive From the aerial parts of C. arboreous, Aguilar-
Guadarrama and Rios30 obtained four sesquiterpenes with
anti-inflammatory activity against ear edema in mice. The Traditional uses, Chemistry and Pharmacology of Croton species 26 J. Braz. Chem. Soc. 7. Concluding Remarks: Points Deserving
Further Work zehntneri
Nervous disturbances, anorexia,
Volatile oil → intestinal muscle relaxant,28,29 depressor central effect,162
gastrointestinal disturbances, sweetener
antinociceptive;6 anethole and estragole → intestinal muscle relaxant;28
anethole → anti-carcinogenic186 Table 1. Species of Croton, traditional uses and pharmacological effects reported for plant parts and some important isola
treated in the text are not included for lack of information of either traditional uses or pharmacological assays Bark volatile oil → gastric ulcer healing,95,145 anti-leishmanial;97 bark water extract →
lowering of the weight gain and higher sensitivity of adipocytes to isoprenaline
and adrenaline;11 trans-crotonin, trans-dehydrocrotonin, acetyl aleuritolic acid →
hypolipidemic and hypoglycaemic effects;50,163 trans-dehydrocrotonin →
anti-oestrogen, anticancer;51 linalool → anti-bacterial and antifungal98
Bark → anti-inflammatory and anti-oxidant12 Aqueous and ethanolic extracts;23,156 quercetin-3,7-dimethyl ether → vasorelaxant
and anti-hypertensive24
Cytotoxic diterpenes66 Plaunotol → anti-peptic ulcer activity,201-203 anticancer,185 anti-bacterial187
Bark red latex → anti-diarrheic,122 antifungal;148 catechin, acetyl aleuritolic
acid → antibacterial;27 acetyl aleuritolic acid → analgesic25
Leaves → cytotoxicity;37 trachylobane diterpene → cytotoxic37 Volatile oil → intestinal muscle relaxant,28,29 depressor central effect,162
antinociceptive;6 anethole and estragole → intestinal muscle relaxant;28
anethole → anti-carcinogenic186 antimalarial effects of secokaurane diterpenes of C. kongensis, the cytotoxicity of taspine, the hypo-
lipidemic and hypoglycaemic effects of C. urucurana
and the cytotoxicity of trachylobane diterpenes of C. zambesicus (Table 1). Asteraceae) and secretory trichomes (e.g. Lamiaceae),
the latter being regarded as evolutionary more recent.218
This is certainly a point deserving research by plant
anatomists. It has been said above (see section Chemical Affinities)
that, among Croton species containing alkaloids, only C. salutaris is known to possess also diterpenes. It was said
also that most species with volatile oils have not been reported
as diterpene- and alkaloid-bearers. Is this apparent mutual
exclusiveness between classes of secondary metabolites a
reality in Croton or just a delusion resultant of insufficient
sampling? Relationships between the occurrences of classes
of secondary metabolites in Croton are a point deserving
consideration, because it may end up useful in future
prospects of pharmacologically active substances. Another aspect that needs investigation deals with
the secretory structures involved in the production and
accumulation of some Croton extractives, such as latex
and volatile oils. With the exception of a study by
Rudall217 about laticifer distribution in Crotonoideae and
other Euphorbiaceae, nothing has been reported about
structures involved in the production and accumulation
of volatile oils in Croton. In angiosperms, volatile oils
may be found inside oil cells (e.g. Lauraceae), secretory
cavities (e.g. 7. Concluding Remarks: Points Deserving
Further Work tonkinensis (several
traditional uses), C. urucurana (pain), C zambesicus
(fever, malaria, convulsions) and C. zehntneri
(anorexia) (Table 1). On the other hand, experimental
work has uncovered potential uses of Croton chemicals
with no records in the list of popular uses. A few
examples are the cytotoxic, antimycobacterial and 27 Vol. 18, No. 1, 2007 Salatino et al. Table 1. Species of Croton, traditional uses and pharmacological effects reported for plant parts and some important isolated compounds. Several species
treated in the text are not included for lack of information of either traditional uses or pharmacological assays
Species
Traditional uses
Effects of plant parts and relevant isolated components
C. arboreous
Anti-inflammatory
Four sesquiterpenes → anti-inflammatory30
C. cajucara
Diabetes, hypercholesterolemia, digestive
Bark volatile oil → gastric ulcer healing,95,145 anti-leishmanial;97 bark water extract →
disturbances, hepatic disturbances,
lowering of the weight gain and higher sensitivity of adipocytes to isoprenaline
weight-loss
and adrenaline;11 trans-crotonin, trans-dehydrocrotonin, acetyl aleuritolic acid →
hypolipidemic and hypoglycaemic effects;50,163 trans-dehydrocrotonin →
anti-oestrogen, anticancer;51 linalool → anti-bacterial and antifungal98
C. celtidifolius
Inflammation, leukaemia,
Bark → anti-inflammatory and anti-oxidant12
ulcers, rheumatism
C. eluteria
Bronchitis, fever, malaria, digestive,
Extracts of bitter bark → stimulation of gastric secretion141
hipertension
C. kongensis
Dysmenorrhea
Secokaurane diterpenes → cytotoxic, anti-mycobacterial and anti-malarial38
C. lechleri
Haemostatic, wound-healing, purgative
Red latex → anti-inflammatory,9 anti-viral,124 antibacterial,142 anti-leukemic;151
SP-303 → anti-viral: respiratory syncytial,191 genital and anogenital simplex lesions,192
C. macrostachys
Purgative, diabetes
Seeds and roots → purgatives35
C. malambo
Pain, rheumatism, inflammation,
Bark extract → antinociceptive, anti-inflammatory161
diarrhea, diabetes, gastric ulcer
C. nepetaefoilius
Stomachic, flatulence, intestinal colic,
Volatile oil → antispasmodic;91,155,204 cineole, methyleugenol → myorelaxant
appetizer
and antispasmodic155
C. oblongifolius
Many applications, including liver
Shoot extract → anti-hepatotoxic activity;41 diterpenes → cytotoxic39,70-73
enlargement, fever and flat worms
C. palanostigma
Intestinal inflammation, wound-healing,
Taspine → cytotoxic110,142
gastric ulcers
C. schiedeanus
Hypertension
Aqueous and ethanolic extracts;23,156 quercetin-3,7-dimethyl ether → vasorelaxant
and anti-hypertensive24
C. tonkinensis
Stomach-ache, dyspepsia, gastric and
Cytotoxic diterpenes66
duodenal ulcers, leprosy, psoriasis,
urticaria
C. sublyratus
Anthelmintic and dermatologic problems
Plaunotol → anti-peptic ulcer activity,201-203 anticancer,185 anti-bacterial187
C. urucurana
Pain reliever, inflammation, wound
Bark red latex → anti-diarrheic,122 antifungal;148 catechin, acetyl aleuritolic
infection, wound-healing, cancer
acid → antibacterial;27 acetyl aleuritolic acid → analgesic25
C. zambesicus
Fever, malaria, convulsions, hypertension,
Leaves → cytotoxicity;37 trachylobane diterpene → cytotoxic37
microbial infections, dysentery
C. Acknowledgments 1. Sampson, J. H.; Phillipson, J. D.; Bowery, N. G.; O’Neill, M. J.; Houston, J. G.; Lewis, J. A.; Phytother. Res. 2000, 14, 24. 1. Sampson, J. H.; Phillipson, J. D.; Bowery, N. G.; O’Neill, M. AS and MLFS are fellow researchers of CNPq (Conselho
Nacional do Desenvolvimento Científico e Tecnológico,
Brasil). J.; Houston, J. G.; Lewis, J. A.; Phytother. Res. 2000, 14, 24. 2. Pereira, A. S.; Carbonell, S. A.; Aquino Neto, F. R.; Fernandes
D. O.; Amaral, A. C. F.; Barnes, R. A.; J. Chromatogr. A 2002,
947, 254. Antonio Salatino got his Ph.D. from
the University of São Paulo in 1979
and carried out postdoctoral programs
with Drs. Tom Mabry at the University
of Texas and David E. Giannasi at the
University of Georgia. He is currently
Full Professor at the Department of
Botany (which he headed in 2001-2004) of the Institute of
Biosciences of the University of São Paulo. His research
interests are chemotaxonomy and applied botany, including
studies about the effects of pollution, analysis of seed oils,
medicinal plants and propolis, and more recently molecular
phylogeny and gene expression. Author of nearly 100 papers
in national and international scientific journals. 3. Block, S.; Baccelli, C.; Tinant, B.; Meervelt, L. C.; Rozenberg,
R.; Jiwan, J-L. H.; Llabrès, G.; Pauw-Gillet, M-C.; Quetin-
Leclercq, J.; Phytochemistry 2004, 65, 1165. 3. Block, S.; Baccelli, C.; Tinant, B.; Meervelt, L. C.; Rozenberg, R.; Jiwan, J-L. H.; Llabrès, G.; Pauw-Gillet, M-C.; Quetin- R.; Jiwan, J-L. H.; Llabrès, G.; Pauw-Gillet, M-C.; Quetin-
Leclercq, J.; Phytochemistry 2004, 65, 1165. Leclercq, J.; Phytochemistry 2004, 65, 1165. 4. Amaral, A. C. F.; Barnes, R. A.; Phytochemistry 1998, 47, 1445. 5. Milanowski, D. J.; Winter, R. E. K..; Elvin-Lewis, M. P. F.; Lewis, W. H.; J. Nat. Prod. 2002, 65, 814. 6. Oliveira, A. C.; Leal-Cardoso, J. H.; Santos, C. F.; Morais, S. M.; Coelho-de-Souza, A. N.; Braz. J. Med. Biol. Res. 2001, 34,
1471. 7. Lopes, D.; Bizzo, H. R.; Sobrinho, A. F. S.; Pereira, M. V. G.;
J. Essent. Oil Res. 2003, 15, 48. 8. Sandoval, M.; Okuhama, N. N.; Clark, M.; Angeles, F. M.; Lao, 8. Sandoval, M.; Okuhama, N. N.; Clark, M.; Angeles, F. M.; Lao,
J.; Bustamante, S.; Miller, M. J.; J. Ethnopharmacol. 2002, 80,
121. Maria Luiza Faria Salatino received
her Ph.D. from the University of São
Paulo and completed postdoctoral
programs with Drs. Tom Mabry and
David E. Giannasi. Acknowledgments Her research deals
with the analysis of plant secondary
metabolites to explore their taxonomic
and economic potential, and analysis of DNA with interests
in molecular systematics. Presently, she is a faculty member
at the Institute of Biosciences, Department of Botany and
head of its Laboratory of Phytochemistry and Molecular
Systematics. Author of 40 papers, including book chapters
and articles of national and international scientific journals. 9. Risco, E.; Ghia, F.; Vila, R.; Iglesias, J.; Álvarez, E.; Canigueral,
S.; Planta Med. 2003, 69, 785. 10. Campos, A. R.; Albuquerque, F. A. A.; Rao, V. S. N.; Maciel, M. A. M.; Pinto, A. C.; Fitoterapia 2002, 73, 116. 11. Grassi-Kassisse, D. M.; Wolf-Nunes, V.; Miotto, A. M.; Farias- 11. Grassi-Kassisse, D. M.; Wolf-Nunes, V.; Miotto, A. M.; Farias-
Silva, E.; Souza-Brito, A. R. M.; Nunes, D. S.; Spadari-
Bratfisch, R. C.; J. Pharm. Pharmacol. 2003, 55, 253. 12. Nardi, G. M.; Felippi, R.; Dalbó, S.; Siqueira-Junior, J. M.;
Arruda, D. C.; Delle Monache, F.; Timbola, A. K.; Pizzolatti, 12. Nardi, G. M.; Felippi, R.; Dalbó, S.; Siqueira-Junior, J. M.;
Arruda, D. C.; Delle Monache, F.; Timbola, A. K.; Pizzolatti,
M. G.; Ckless, K.; Ribeiro-do-Vale, R. M.; Phytomedicine 2003, M. G.; Ckless, K.; Ribeiro-do-Vale, R. M.; Phytomedicine 2003,
10, 176. 13. Vigor, C.; Fabre, N.; Fourasté, I.; Moulis, C.; Phytochemistry
2001, 57, 1209. 2001, 57, 1209. Giuseppina Negri obtained a Ph. D. in Organic Chemistry from the State
University of Campinas (UNICAMP). She gained expertise in organic
synthesis and mass spectroscopy
working under the supervision of Prof. Dra. Concetta Kascheres. She held a
post-doctorate position at the University of São Paulo,
Department of Botany, when she gained familiarity with
analysis of natural products. GN has carried out
investigations on plant secondary metabolites, with
emphasis in analysis of constituents of medicinal plants
and propolis. Presently, she integrates the group led by Dr. Elisaldo Luiz de Araujo Carlini at CEBRID/UNIFESP,
being involved in several projects dealing with medicinal
plants with activity on the central nervous system. Author
of 30 papers, including book chapters and articles of
national and international scientific journals. 14. Duke, J.; Vasquez, R.; Amazonian Ethnobotanical Dictionary,
CRC Press: Boca Raton, 1994. 15. Cai, Y.; Evans, F.; Roberts, M.; Phillipson, J. D.; Zenk, M.;
Gleba, Y.; Phytochemistry 1991, 30, 2033. 16. Cai, Y.; Chen, Z. P.; Phillipson, J. D.; Phytochemistry 1993,
32, 755. 17. Cai, Y.; Chen, Z. P.; Phillipson, J. 7. Concluding Remarks: Points Deserving
Further Work Myrtaceae), secretory ducts (e.g. J. Braz. Chem. Soc. Traditional uses, Chemistry and Pharmacology of Croton species 28 Acknowledgments J.; Echevarria, A.; Grynberg, N. F.; Côlus, I. M. S.; Farias, R. A. F.; Costa, A. M. L.; Rao, V. S. N.; J. Ethnopharmacol. 2000, 70, 41. 51. Grynberg, N. F.; Echevarria, A.; Lima, J. E.; Pamplona, S. S. 51. Grynberg, N. F.; Echevarria, A.; Lima, J. E.; Pamplona, S. S. R Pi t
A C M
i l M A M Pl
t M d 1999 65 687 51. Grynberg, N. F.; Echevarria, A.; Lima, J. E.; Pamplona, S. S. R.; Pinto, A. C.; Maciel, M. A. M.; Planta Med. 1999, 65, 687. 27. Peres, M. T. L. P.; Delle Monache, F.; Bella Cruz, A.; Pizzolatti,
M. G.; Yunes, R. A.; J. Ethnopharmacol. 1997, 56, 223. R.; Pinto, A. C.; Maciel, M. A. M.; Planta Med. 1999, 65, 687. 52. Vigor, C.; Fabre, N.; Fourasté, I.; Moulis, C.; J. Nat. Prod. 2002, 65, 1180. 28. Coelho-de-Souza, A. N.; Barata, E. L.; Magalhães, P. J. C.;
Lima, C. C.; Leal-Cardoso, J. H.; Phytother. Res. 1997, 11,
299. 53. Fattorusso, E.; Tagliatela-Scafati, O.; Campagnuolo, C.;
Santelia, F. U.; Appendino, G.; Spagliardi, P.; J. Agric. Food
Chem. 2002, 50, 5131. 29. Coelho-de-Souza, A. N.; Criddle, D. N.; Leal-Cardoso, J. H.;
Phytother. Res. 1998, 12, 189. 30. Aguilar-Guadarrama, A. B.; Rios, M. Y.; J. Nat. Prod. 2004,
67, 914. 54. Costa, M.; Perles, E. C.; Fujiwara, F. Y.; Imamura, P. M.;
Phytochemistry 2000, 53, 851. 31. Williams, L.; Evans, P. E.; Bowers, W. S.; J. Chem. Ecol. 2001,
27, 203. 55. Wilson, S. R.; Neubert, L.; Huffman, J. C.; J. Am. Chem. Soc. 1976, 98, 3669. 32. Chavez, P. I.; Jolad, S. D.; Hoffmann, J. J.; Cole, J. R.; J. Nat. Prod. 1982, 45, 745. 56. Puebla, P.; López, J. L.; Guerrero, M.; Carrón, R.; Martín, M. L.; San Román, L.; San Feliciano, A.; Phytochemistry 2003,
62, 551. 33. Murillo, R. M.; Jakupovic, J.; Rivera, J.; Castro, V. H.; Rev. Biol. Trop. 2001, 49, 259. 57. Guerrero, M. F.; Puebla, P.; Carrón, R.; Martín, M. L.; San
Román, L.; J. Ethnopharmacol. 2004, 94, 185. 34. Kapingu, M. C.; Guillaume, D.; Mbwambo, Z. H.; Moshi, M. J.; Uliso, F. C.; Mahunnah, R. L. A.; Phytochemistry 2000, 54,
767. 58. Puebla, P.; Correa, S. X.; Guerrero, M.; Carrón, R.; San
Feliciano, A.; Chem. Pharm. Bull. 2005, 53, 328. 35. Mazzanti, G.; Bolle, P.; Martinoli, L.; Piccinelli, D.; Grgurina,
I.; Animati, F.; Mugnê, Y.; J. Ethnopharmacol. Acknowledgments D.; Phytochemistry 1993,
34, 265. 18. Suárez, A. I.; Compagnone, R. S.; Salazar-Bookaman, M. M.;
Tillet, S.; Delle Monache, F.; Di Giulio, C.; Bruges, G.; J. Ethnopharmacol. 2003, 88, 11. 19. Moura, V. L. A.; Monte, F. J. O.; Braz-Filho, R.; J. Nat. Prod. 1990, 53, 1566. 20. Lahlou, S.; Leal-Cardoso, J. H.; Magalhães, P. J. C.; Planta
Med. 2000, 66, 138. 21. Porrasreyes, R. H.; Lewis, W. H.; Roman, J.; Simchowitz, L.;
Mustoe, T. A.; Proc. Soc. Exp. Biol. Med. 1993, 203, 18. Vol. 18, No. 1, 2007 Salatino et al. 29 45. Vongchareonsathit, A.; De-Eknamkul, W.; Planta Med. 1998,
64, 279. 22. Miller, M. J. S.; MacNaughton, W. K.; Zhang, X-J.; Thompson,
J. H.; Charbonnet, R. M.; Bobrowski, P.; Lao, J.; Trentacosti,
A. M.; Sandoval, M.; Am. J. Physiol.-Gastroint. Liver Physiol. 2000, 279, G192. 46. Tsai, J. C.; Tsai, S. L.; Chang, W. C.; Biol. Pharm. Bull. 2004,
27, 162. 23. Guerrero, M. F.; Carrón, R.; Martin, M. L.; San Román, L.;
Reguero, M. T.; J. Ethnopharmacol. 2001, 75, 33. 47. Giang, P. M.; Jin, H. Z.; Son, P. T.; Lee, J. H.; Hong, Y. S.; Lee,
J. J.; J. Nat. Prod. 2003, 66, 1217. 48. Maciel, M. A. M.; Pinto, A. C.; Kaiser, C. R.; Magn. Reson. Chem. 2003, 41, 278. 24. Guerrero, M. F.; Puebla, P.; Carrón, R.; Martin, M. L.; Román,
L. S.; J. Pharm. Pharmacol. 2002, 54, 1373. 25. Peres, M. T. L. P.; Pizzolatti, M. G.; Yunes, R. A.; Delle
Monache, F.; Phytochemistry 1998, 49, 171. 49. Maciel, M. A. M.; Pinto, A. C.; Brabo, S. N.; Silva, M. N.;
Phytochemistry 1998, 49, 823. 50. Maciel, M. A. M.; Pinto, A. C.; Arruda, A. C.; Pamplona, S. G. 50. Maciel, M. A. M.; Pinto, A. C.; Arruda, A. C.; Pamplona, S. G. S. R.; Vanderlinde, F. A.; Lapa, A. J.; Echevarria, A.; Grynberg, 50. Maciel, M. A. M.; Pinto, A. C.; Arruda, A. C.; Pamplona, S. G. S. R.; Vanderlinde, F. A.; Lapa, A. J.; Echevarria, A.; Grynberg,
N. F.; Côlus, I. M. S.; Farias, R. A. F.; Costa, A. M. L.; Rao, V. S. N.; J. Ethnopharmacol. 2000, 70, 41. 26. Orlandi-Mattos, P. E.; Geremias, R.; Cordova, C. A. S.;
Creczynski-Pasa, T. B.; Rebello, J. M.; Wilhelm, D.; Martins,
D. T. O.; Llesuy, S.; Pedrosa, R. C.; Free Radical Biol. Med. 2002, 33, 645. S. R.; Vanderlinde, F. A.; Lapa, A. Acknowledgments 99. Agnaniet, H.; Akagah, A.; Mounzeo, H.; Menut, C.; Bessiere,
J.M.; J. Essent. Oil Res. 2005, 17, 201. J.M.; J. Essent. Oil Res. 2005, 17, 201. 77. Hernandez, J.; Delgado, G.; Fitoterapia 1992, 63, 377. 100. Martins, A.P.; Salgueiro, L. R.; Gonçalves, M. J.; Vila, R.; Tomi,
F.; Adzet, T.; da Cunha, A. P.; Canigueral, S.; Casanova, J.;
Planta Med. 2000, 66, 647. 78. Ngamrojanavanich, N.; Tonsiengsom, S.; Lertpratchya, P.;
Roengsumran, S.; Puthong, S.; Petsom, A.; Arch. Pharm. Res. 2003, 26, 898. 101. Dourado, R. C. M.; Silveira, E. R.; J. Essent. Oil Res. 2005,
17, 36. 79. Baccelli, C.; Block, S.; Van Holle, B.; Schanck, A.; Chapon,
D.; Tinant, B.; Van Meervelt, L.; Morel, N.; Quetin-Leclerog,
J.; Planta Med. 2005, 71, 1036. 102. Catalán, C. A. N.; Heluani, C. S.; Kotowicz, C.; Gedris, T. E.;
Herz, W.; Phytochemistry 2003, 64, 625. 80. Sutthaivaiyakit, S.; Nareeboon, P.; Ruangrangsi, N.;
Ruchirawat, S.; Pisutjaroenpong, S.; Mahidol, C.;
Phytochemistry 2001, 56, 811. 103. Palmeira, S. F.; Moura, F. D.; Alves, V. D.; De Oliveira, F. M.;
Bento, E. S.; Conserva, L. M.; Andrade, E. H. D.; Flavour Fragr. J. 2004, 19, 69. 104. Meccia, G.; Rojas, L. B.; Rosquete, C.; San Feliciano, A.;
Flavour Fragr. J. 2000, 15, 144. 81. Graikou, K.; Aligiannis, N.; Skaltsounis, A. L.; Chinou, L.;
Michel, S.; Tillequin, F. O.; Litaudon, M.; J. Nat. Prod. 2004,
67, 685. 105. Block, S.; Flamini, G.; Brkic, D.; Morelli, I.; Quetin-Leclercq,
J.; Flavour Fragr. J. 2006, 21, 222. 82. Graikou, K.; Aligiannis, N.; Chinou, L.; Skaltsounis, A. L.; 82. Graikou, K.; Aligiannis, N.; Chinou, L.; Skaltsounis, A. L.;
Tilleguin, F. O.; Litaudon, M.; Helv. Chim. Acta 2005, 88, 2654. 106. Barbosa, P. R.; Fascio, M.; Martins, D.; Guedes, M. L. S.;
Roque, N. F.; Biochem. Syst. Ecol. 2003, 31, 307. Tilleguin, F. O.; Litaudon, M.; Helv. Chim. Acta 2005, 88, 2654. 83. Itokawa, H.; Ichihara, Y.; Takeya, K.; Morita, H.; Motidome,
M.; Phytochemistry 1991, 30, 4071. 107. Peres, M. T. L. P.; Delle Monache, F.; Pizzolatti, M. G.; Santos,
A. R. S.; Beirith, A..; Calixto, J. B.; Yunes, R. A.; Phytother. Res. 1998, 12, 209. 84. Alexander, I. C.; Pascoe, K. O.; Manchard, P.; Williams, L. A. D.; Phytochemistry 1991, 30, 1801. 85. de Heluani, C. S.; Catalan, C. A. N.; Hernandez, L. R.;
Burgueno-Tapia, E.; Joseph-Nathan, P.; Magn. Reson. Chem. 1998, 36, 947. 108. Acknowledgments C.; Criddle, D. N.; Tavares, R. A.; Melo, E. M.;
Mota, T. L.; Leal-Cardoso, J. H.; Phytother. Res. 1998, 12, 172. 92. Batatinha, M. J. M.; Souza-Spinosa, H.; Bernardi, M. M.; J. Ethnopharmacol. 1995, 45, 53. 71. Roengsumran, S.; Petsom, A.; Kuptiyanuwat, N.; Vilaivan, T.;
Ngamrojanavanich, N.; Chaichantipyuth, C.; Puthong, S.;
Phytochemistry 2001, 56, 103. 93. Ciccio, J.F.; Segnini, M.; J. Essent. Oil Res. 2002, 14, 357. 94. Suárez, A. I.; Vásquez, L. J.; Manzano, M. A.; Compagnone,
R. S.; Flavour Fragr. J. 2005, 20, 611. 94. Suárez, A. I.; Vásquez, L. J.; Manzano, M. A.; Compagnone, R. S.; Flavour Fragr. J. 2005, 20, 611. 72. Roengsumran, S.; Musikul, K.; Petsom, A.; Vilaivan, T.;
Sangvanich, P.; Pompakakul, S.; Puthong, S.; Chaichantipyuth,
C.; Jaiboon, N.; Chaichit, N.; Planta Med. 2002, 68, 274. R. S.; Flavour Fragr. J. 2005, 20, 611. 95. Hiruma-Lima, C. A.; Gracioso, J. S.; Nunes, D. S.; Souza-Brito,
A. R. M.; J. Pharm. Pharmacol. 1999, 51, 341. 95. Hiruma-Lima, C. A.; Gracioso, J. S.; Nunes, D. S.; Souza-Brito, A. R. M.; J. Pharm. Pharmacol. 1999, 51, 341. C.; Jaiboon, N.; Chaichit, N.; Planta Med. 2002, 68, 27 96. Lopes, D.; Bizzo, H. R.; Sobrinho, A. F. S.; Pereira, M. V. G.;
J. Essent. Oil Res. 2000, 12, 705. 73. Roengsumran, S.; Pompakakul, S.; Muangsin, N.; Sangvanich,
P.; Nhujak, T.; Singtothong, P.; Chaichit, N.; Puthong, S.;
Petsom, A.; Planta Med. 2004, 70, 87. J. Essent. Oil Res. 2000, 12, 705. 97. Rosa, M. D. S.; Mendonça, R. R.; Bizzo, H. R.; Rodrigues, I. 97. Rosa, M. D. S.; Mendonça, R. R.; Bizzo, H. R.; Rodrigues, I. D.; Soares, M. A.; Souto Padron, T.; Alviano, C. S.; Lopes, A. H. C. S.; Antimicrob. Agents Chemother. 2003, 47, 1895. 74. Chaichantipyuth, C.; Petsom, A.; Taweechotipatr, P.; Muangsin,
N.; Chaichit, N.; Puthong, S.; Roengsumran, S.; Kawahata, M.;
Watanabe, T.; Ishikawa, T.; Heterocycles 2005, 65, 809. N.; Chaichit, N.; Puthong, S.; Roengsumran, S.; Kawahata, Watanabe, T.; Ishikawa, T.; Heterocycles 2005, 65, 809 98. Alviano, W. S.; Mendonça, R. R.; Alviano, D. S.; Bizzo, H. R.;
Souto-Padron, T.; Rodrigues, M. L.; Bolognese, A. M.; Alviano,
C. S.; Souza, M. M. G.; Oral Microbiol. Immunol. 2005, 20, 101. 75. Luzbetak, D. J.; Torrance, S. J.; Hoffmann, J. J.; Cole, J. R.; J. Nat. Prod. 1979, 42, 315. 76. Bandara, B. M. R.; Wimalasiri, W. R.; Bandara, K. A. N. P.;
Planta Med. 1987, 6, 575. Acknowledgments 1987, 19, 213. 35. Mazzanti, G.; Bolle, P.; Martinoli, L.; Piccinelli, D.; Grgurina, 59. Palmeira, S. F.; Conserva, L. M.; Silveira, E. R.; J. Braz. Chem. Soc. 2005, 16, 1420. I.; Animati, F.; Mugnê, Y.; J. Ethnopharmacol. 1987, 19, 213. 60. Ngadjui, B. T.; Folefoc, G. N.; Keumedjio, F.; Dengo, E.;
Sondengam, B. L.; Connolly, J. D.; Phytochemistry 1999, 51, 171. 36. Ngadjui, B. T.; Abegaz, B. M.; Keumedjio, F.; Folefoc, G. N.;
Kapche, G. W. F.; Phytochemistry 2002, 60, 345. 61. Krebs, H. C.; Ramiarantsoa, H.; Phytochemistry 1996, 41, 561. 37. Block, S.; Stévigny, C.; De Pauw-Gillet, M. C.; De Hoffmann,
E.; Llabrés, G.; Adjakidjé, V.; Quetin-Leclercq, J.; Planta Med. 2002, 68, 647. 62. Krebs, H. C.; Ramiarantsoa, H.; Phytochemistry 1997, 45, 379. 63. Son, P. T.; Giang, P. M.; Taylor, W. C.; Aust. J. Chem. 2000, 53,
1003. 38. Thongtan, J.; Kittakoop, P.; Ruangrungsi, N.; Saenboonrueng,
J.; Thebtaranonth, Y.; 2003. J. Nat. Prod. 2003, 66, 868. 64. Minh, P. T. H.; Ngoc, P. H.; Quang, D. N.; Hashimoto, T.;
Takaoka, S.; Asakawa, Y.; Chem. Pharm. Bull. 2003, 51, 590. 39. Sommit, D.; Petsom, A.; Ishikawa, T.; Roengsumran, S.; Planta
Med. 2003, 69, 167. 65. Giang, P. M.; Son, P. T.; Lee, J. J.; Otsuka, H.; Chem. Pharm. Bull. 2004, 52, 879. 40. Ngamrojnavanich, N.; Sirimongkon, S.; Roengsumran, S.;
Petsom, A.; Kamimura, H.; Planta Med. 2003, 69, 555. 66. Giang, P. M.; Son, P. T.; Hamada, Y.; Otsuka, H.; Chem. Pharm. Bull. 2005, 53, 296. 41. Ahmed, B.; Alam, T.; Varshney, M.; Khan, S. A.; J. Ethnopharmacol. 2002, 79, 313. 67. Wungsintaweekul, J.; De-Eknamkul, W.; Tetrahedron Lett. 2005, 46, 2125. 42. Singh, M.; Pal, M.; Sharma, R. P.; Planta Med. 1999, 65, 2. 68. Aiyar, V. N.; Seshadri, T. R.; Phytochemistry 1972, 11, 1473. 43. Gupta, M.; Mazumber, U. K.; Vanisi, M. L. M.; Sivakumar, T.;
Kandar, C. C.; J. Ethnopharmacol. 2004, 90, 21. 69. Roengsumran, S.; Petsom, A.; Sommit, D.; Vilaivan, T.;
Phytochemistry 1999, 50, 449. 44. Tansakul, P.; De-Eknamkul, W.; Phytochemistry 1998, 47, 1241. Traditional uses, Chemistry and Pharmacology of Croton species 30 J. Braz. Chem. Soc. 91. Magalhães, P. J. C.; Criddle, D. N.; Tavares, R. A.; Melo, E. M.;
Mota, T. L.; Leal-Cardoso, J. H.; Phytother. Res. 1998, 12, 172. 70. Roengsumran,
S.;
Singtothong,
P.;
Pudhom,
K.;
Ngamrojanavanich, N.; Petsom, A.; Chaichantipyuth, C.; J. Nat. Prod. 1999, 62, 1163. 91. Magalhães, P. J. Acknowledgments Rossi, D.; Bruni, R.; Bianchi, N.; Chiarabelli, C.; Gambari, R.;
Medici, A.; Lista, A.; Paganetto, G.; Phytomedicine 2003, 10,
139. 128. González-Vázquez, R.; Diaz, B. K.; Aguilar, M. I.; Diego, N.;
Lotina-Hennsen, B.; J. Agric. Food Chem. 2006, 54, 1217. 152. Lopes, M. I. L. E.; Saffi, J.; Echeverrigaray, S.; Henriques, J. A. N.; Salvador, M.; J Ethnopharmacol. 2004, 95, 437. 129. Markham, K.R.; Techniques of Flavonoid Identification,
Academic Press: London, 1982. 153. Santos, F. V.; Mesquita, S. F. P.; Faria, M. J. S. S.; Poersh, A.;
Maciel, M. A. M.; Pinto, A. C.; Morimoto, H. K.; Colus, I. M. S.; Genet. Mol. Biol. 2006, 29, 159. 130. Athikomkulchai, S.; Prawat, H.; Thasana, N.; Ruangrungsi, N.;
Ruchirawat, S.; Chem. Pharm. Bull. 2006, 54, 262. 154. Tieppo, M.; Porawski, M.; Salvador, M.; Moreira, A. J.;
Collado, P. S.; Gonzalez-Gallego, J.; Marroni, N. P.; Biol. Pharm. Bull. 2006, 29, 161. 131. Pieters, L.; De Bruyne, T.; Claeys, M.; Vlietinck, A.; Calomme, 131. Pieters, L.; De Bruyne, T.; Claeys, M.; Vlietinck, A.; Calomme,
M.; Berghe, D. V.; J. Nat. Prod. 1993, 56, 899. M.; Berghe, D. V.; J. Nat. Prod. 1993, 56, 899. 155. Magalhães, P. J. C.; Lahlou, S.; Vasconelos Dos Santos, M. A.;
Pradines, T. L.; Leal-Cardoso, J. H.; Planta Med. 2003, 69, 874. 132. Puebla, P.; Correa, S. X.; Guerrero, R.; Feliciano, A. S.;
Biochem. Syst. Ecol. 2005, 33, 849. 156. Lahlou, S.; Leal-Cardoso, J. H.; Magalhães, P. J. C.; Coelho-
de-Souza, A. N.; Pinto-Duarte, G.; Planta Med. 1999, 65, 553. 133. Abe, I.; Takahashi, Y.; Morita, H.; Noguchi, H.; Eur. J. Biochem. 2001, 268, 3354. 157. Guerrero, M. F.; Puebla, P.; Carrón, R.; Martin, M. L.; Arteaga, 157. Guerrero, M. F.; Puebla, P.; Carrón, R.; Martin, M. L.; Arteaga,
L.; Román, L. S.; J. Ethnopharmacol. 2002, 80, 37. 134. Pereira, A. S.; Amaral, A. C. F.; Barnes, R. A.; Cardoso, F. R.;
Aquino Neto, F. R.; Phytochem. Anal. 1999, 10, 254. 134. Pereira, A. S.; Amaral, A. C. F.; Barnes, R. A.; Cardoso, F. R.; 157. Guerrero, M. F.; Puebla, P.; Carrón, R.; Martin, M. L.; Arteaga,
L.; Román, L. S.; J. Ethnopharmacol. 2002, 80, 37. Aquino Neto, F. R.; Phytochem. Anal. 1999, 10, 254. L.; Román, L. S.; J. Ethnopharmacol. 2002, 80, 37. 35. Mukherjee, R.; Axt, E. M.; Phytochemistry 1984, 23, 2 158. Abdon, A. P. V.; Leal-Cardoso, J. H.; Coelho-de-Souza, A. N.;
Morais, S. Acknowledgments Tsacheva, I.; Rostan, J.; Iossifova, T.; Vogler, B.; Odjakova,
M.; Navas, H.; Kostova, I.; Kojoharova, M.; Kraus, W.; Z. Naturforsch., C: J. Biosci. 2004, 59C, 528. 86. de Heluani, C. S.; Catalan, C. A. N.; Hernandez, L. R.;
Burgueno-Tapia, E.; Joseph-Nathan, P.; J. Nat. Prod. 2000, 63,
222. 109. Vaisberg, A. J.; Milla, M.; Planas, M. D.; Cordova, J. L.;
Deagusti E. R.; Ferreyra, R.; Mustiga, M. D.; Carlin, L.;
Hammond, G. B.; Planta Med. 1989, 55, 140. 87. Evans, F. J. In Naturally Occurring Phorbol Esters; Evans, E. J., ed., CRC Press: Boca Raton, 1986, p. 171. 110. Itokawa, H.; Ichihara, Y.; Mochizuki, M.; Enomori, T.; Morita,
H.; Shirota, O.; Inamatsu, M.; Takeya, K.; Chem. Pharm. Bull. 1991, 39, 1041. J., ed., CRC Press: Boca Raton, 1986, p. 171. 88. Glaser, S.; Sorg, B.; Hecker, E. A.; Planta Med. 1988, 54, 580. 111. Carlin, L.; Vaisberg, A. J.; Hammond, G. B.; Planta Med. 1996,
62, 90. 89. El-Mekkawy, S.; Meselly, M. R.; Nakamura, N.; Hattori, M.;
Kawahata, T.; Otake, T.; Chem. Pharm. Bull. 1999, 47, 1346. 112. Eisenreich, W. J.; Höfner, G.; Bracher, F.; Nat. Prod. Res. 2003,
17, 437. 90. El-Mekkawy, S.; Meselly, M. R.; Nakamura, N.; Hattori, M.;
Kawahata, T.; Otake, T.; Phytochemistry 2000, 53, 457. Vol. 18, No. 1, 2007 31 Salatino et al. 113. Charris, J.; Dominguez, J.; De La Rosa, C.; Caro, C.; Biochem. Syst. Ecol. 2000, 28, 795. 140. Farias, R. A. F.; Rao, V. S. N.; Viana, G. S. B.; Silveira, E. R.; Maciel, M. A. M.; Pinto, A. C.; Planta Med. 1997, 66,
558. 114. Barnes, R. A.; Soeiro, O. M.; Phytochemistry 1981, 20, 543. 115. Bruneton, J.; Pharmacognosy, Phytochemistry, Medicinal
Plants, Intercept Ltd.: London, 1999. 141. Appendino, G.; Borrelli, F.; Capasso, R.; Campagnuolo, C.;
Fattorusso, E.; Petrucci, F.; Taglialatela-Scafati, O.; J. Agric. Food Chem. 2003, 51, 6970. 116. Amaral, A. C. F.; Barnes, R. A.; Planta Med. 1997, 63, 485. 142. Chen, Z-P.; Cai, Y.; Phillipson, J. D.; Planta Med. 1994, 60, 541. 117. Stuart, K. L.; Graham, L.; Phytochemistry 1973, 12, 1973. 143. Jones, K.; J. Altern. Complem. Med. 2003, 9, 877. 118. Araujo-Junior, V. T.; da Silva, M. S.; da Cunha, E. V. L.; Agra, 118. Araujo-Junior, V. T.; da Silva, M. S.; da Cunha, E. V. L.; Agra,
M. D.; da Silva, R. N.; Barbosa, J. M.; Braz-Filho, R.; Pharm. Biol. 2004, 42, 62. 144. Hiruma-Lima, C. Acknowledgments A.; Gracioso, J. S.; Rodriguez, J. A.; Haun,
M.; Nunes, D. S.; Souza-Brito, A. R. M.; J. Ethnopharmacol. 2000, 69, 229. 119. Suárez, A. I.; Suleyma, B.; Delle Monache, F.; Compagnone,
R S ; Arvelo F ; Nat Prod Res 2004 18 421 119. Suárez, A. I.; Suleyma, B.; Delle Monache, F.; Compagnone, 119. Suárez, A. I.; Suleyma, B.; Delle Monache, F.; Compagnone,
R. S.; Arvelo, F.; Nat. Prod. Res., 2004, 18, 421. R. S.; Arvelo, F.; Nat. Prod. Res., 2004, 18, 421. 145. Hiruma-Lima, C. A.; Gracioso, J. S.; Bighetti, E. J.; Grassi-
Kassisse, D. M.; Nunes, D. S.; Souza-Brito, A. R. M.;
Phytomedicine 2002, 9, 523. 120. Araujo-Junior, V. T.; da Silva, M. S.; da Cunha, E. V. L.; Agra,
M. D.; Athayde-Filho, P. F.; Vieira, I. J. C.; Braz-Filho, R.;
Barbosa-Filho, J. M.; J. Braz. Chem. Soc. 2005, 16, 553. 146. Fischer, H.; Machen, T. E.; Widdicombe, J. H.; Carlson, T. J. S.; King, S. R.; Chow, J. W. S.; Illek, B.; J. Ethnopharmacol. 2004, 93, 351. Barbosa-Filho, J. M.; J. Braz. Chem. Soc. 2005, 16, 553. 121. Pieters, L.; De Bruyne, T.; Van Poel, B.; Vingerhoets, R.; Totté,
J.; Van Den Benghe, D.; Phytomedicine 1995, 2, 17. J.; Van Den Benghe, D.; Phytomedicine 1995, 2, 17. 147. Desmarchelier, C.; Witting Schaus, F.; Coussio, J.; Cicca, G.;
J. Ethnopharmacol. 1997, 58, 103. 122. Gurgel, L. A.; Silva, R. M.; Santos, F. A.; Martins, D. T.; Mattos, P. O.; Rao, V. S.; Phytother. Res. 2001, 15, 319. 123. Wyde, P. R.; Ambrose, M. W.; Meyerson, L. R.; Gilbert B. E.;
Antiviral Res. 1993, 20, 145. 148. Gurgel, L. A.; Sidrim, J. J. C.; Martins, D. T.; Cechinel, V.;
Rao, V. S.; J. Ethnopharmacol. 2005, 97, 409. 124. Ubillas, R.; Phytomedicine 1994, 1, 77. 149. Prozesky, E. A.; Meyer, J. J. M.; Louw, A. I.; J. Ethnopharmacol. 2001, 76, 239. 125. Abo, K. A.; Ogunleye, V. O.; Ashidi, J. S.; Phytother. Res. 1999,
13, 494. 150. Weniger, B.; Lagnika, L.; Vonthron-Sénécheau, C.; Adjobimey,
T.; Gbenou, J.; Moudachirou, M.; Brum, R.; Anton, R.; Sanni,
A.; J. Ethnopharmacol. 2004, 90, 279. 126. Kostova, I.; Iossifova, T.; Rostan, J.; Vogler, B.; Kraus, W.;
Navas, H.; Pharm. Pharmacol. Lett. 1999, 9, 34. 127. Dalbo, S.; Jungensen, S.; Horst, H.; Ruzza, A. A.; Soethe, D. N.; Santos, A. R. S.; Pizzolati, M. G.; Ribeiro-do-Valle, R. M.;
J. Pharm. Pharmacol. 2005, 57, 765. 151. Traditional uses, Chemistry and Pharmacology of Croton species Med. Chem. 1965, 8, 672. 193. Nakamura, N.; Yakugaku Zasshi – J. Pharm. Soc. Japan 2004,
124, 519. 170. Hecker, E.; Bartsch, H.; Bresch, H.; Gshwendt, M.; Härle, E.;
Kreibich, G.; Kubinyl, H.; Schairer, H. U.; Szczepanski, Ch. V.; Thielmann, H. W.; Tetrahedron Lett. 1967, 33, 3165. Hecker, E.; Bartsch, H.; Bresch, H.; Gshwendt, M.; Härle 194. Masuda, T.; Harada, S.; Virology 1993, 192, 696. 171. Castagna, M.; Takai, Y.; Kaibuchi, K.; Sano, K.; Kikkawa, U.;
Nishizuka, Y.; J. Biol. Chem. 1982, 257, 7847. 195. Souza-Brito, A. R. M.; Rodríguez, J. A.; Hiruma-Lima, C. A.;
Haun, M.; Nunes, D. S.; Planta Med. 1998, 64, 126. 195. Souza-Brito, A. R. M.; Rodríguez, J. A.; Hiruma-Lima, C. A.; 195. Souza-Brito, A. R. M.; Rodríguez, J. A.; Hiruma-Lima
Haun, M.; Nunes, D. S.; Planta Med. 1998, 64, 126. Haun, M.; Nunes, D. S.; Planta Med. 1998, 64, 126. Nishizuka, Y.; J. Biol. Chem. 1982, 257, 7847. 196. Hiruma-Lima, C. A.; Spadari-Bratfisch, R. C.; Kassisse, D. M.; 172. Nishizuka, Y.; Science 1992, 258, 607. 196. Hiruma-Lima, C. A.; Spadari-Bratfisch, R. C.; Kassisse, D. M.;
Souza-Brito, A. R. M.; Planta Med. 1999, 65, 325. 173. Aitken, A. In Naturally Occurring Phorbol Esters; Evans, E. J., ed.; CRC Press: Boca Raton, 1986, p. 271. 173. Aitken, A. In Naturally Occurring Phorbol Esters; Evans, E. Souza-Brito, A. R. M.; Planta Med. 1999, 65, 325. 173. Aitken, A. In Naturally Occurring Phorbol Esters;
J., ed.; CRC Press: Boca Raton, 1986, p. 271. 197. Hiruma-Lima, C. A.; Toma, W.; Gracioso, J. S.; Almeida, A. B. A.; Batista, L. M.; Magri, L.; Bensuaski de Paula, A. C.; Soares,
F. R.; Nunes, D. S.; Souza-Brito, A. R. M.; Biol. Pharm. Bull. 2002, 25, 452. J., ed.; CRC Press: Boca Raton, 1986, p. 271. 174. Bertolini, T. M.; Giorgione, J.; Harvey, D. F.; Newton, A. C.; J. Org. Chem. 2003, 68, 5028. 175. Abdel-Hafez, A. A. M.; Nakamura, N.; Hattori, M.; Chem. Pharm. Bull. 2002, 50, 160. 198. Rodriguez, J.A.; Hiruma-Lima, C.A.; Souza-Brito, A. R. M.,
Hum. Exp. Toxicol. 2004, 23, 455. 176. Melo, P. S.; Justo, G. Z.; Durán, N.; Haun, M.; Eur. J. Pharmacol. 2004, 487, 47. 199. Almeida, A. B. A..; Melo, P. S.; Hiruma-Lima, C. A.; Gracioso,
J. S.; Carli, l.; Nunes, D. S.; Haun, M.; Souza-Brito, A. R. M.;
Eur. J. Pharmacol. 2003, 472, 205. 177. Anazetti, M. C.; Melo, P. Traditional uses, Chemistry and Pharmacology of Croton species Traditional uses, Chemistry and Pharmacology of Croton species 32 J. Braz. Chem. Soc. 186. Chainy, G. B. N.; Manna, S. K.; Chaturvedi, M. M.; Aggarwal,
B. B.; Oncogene 2000, 19, 2943. 161. Suárez, A. I.; Compagnone, R. S.; Salazar-Bookaman, M. M.;
Tillet, S.; Delle Monache, F.; Di Giulio, C.; Bruges, G.; J. Ethnopharmacol. 2003, 88, 11. 187. Matsumoto, Y.; Hamashima, H.; Masuda, K.; Shiojima, K.;
Sasatsu M ; Arai T ; Microbios 1998 96 149 187. Matsumoto, Y.; Hamashima, H.; Masuda, K.; Shiojima, K.; 187. Matsumoto, Y.; Hamashima, H.; Masuda, K.; Shiojima, K.;
Sasatsu, M.; Arai, T.; Microbios 1998, 96, 149. 162. Lazarini, C. A.; Uema, A. H.; Brandão, G. M. S.; Guimarães,
A. P. C.; Bernardi, M. M.; Phytomedicine 2000, 7, 477. 188. Inoue, Y.; Shiraishi, A.; Hada, T.; Hirose, K.; Hamashima, H.; 188. Inoue, Y.; Shiraishi, A.; Hada, T.; Hirose, K.; Hamashima, H.;
Shimada, J.; Fems Microbiol. Lett. 2004, 237, 325. Shimada, J.; Fems Microbiol. Lett. 2004, 237, 325. 163. Maciel, M. A. M.; Pinto, A. C.; Veiga Jr., V. F.; Grynberg, N. F.;
Echevarria, A.; Quim. Nova, 2002, 25, 429. 189. de Bruyne, T.; Pieters, L.; Deelstra, H.; Vlietinck, A.; Biochem. Syst. Ecol. 1999, 27, 445. Echevarria, A.; Quim. Nova, 2002, 25, 429. 164. Silva, R. M.; Santos, F. A.; Rao, V. S. N.; Maciel, M. A. M.;
Pinto, A. C.; J. Pharm. Pharmacol. 2001, 53, 535. 190. Wyde, P. R.; Meyerson, L. R.; Gilbert B. E.; Drug Develop. Res. 1993, 28, 467. Pinto, A. C.; J. Pharm. Pharmacol. 2001, 53, 535. 165. Silva, R. M.; Santos, F. A..; Maciel, M. A. M.; Pinto, A. C.;
Rao, V. S.; Planta Med. 2001, 67, 763. 191. Barnard, D. L.; Smee, D. F.; Huffman, J. H.; Meyerson, L. R.;
Sidwell, R. W.; Chemotherapy 1993, 39, 203. 166. Tomei, D. L.; Noyes, I.; Blocker, D.; Holliday, J.; Glaser, R.;
Nature 1987, 329, 73. Sidwell, R. W.; Chemotherapy 1993, 39, 203. 192. Orozco-Topete, R.; Sierra-Madero, J.; Cano Dominguez, C.;
Kershenovich, J.; Ortiz Pedroza, G.; Vazquez Valls, E.; Garcia
Cosio, C.; Soria Córdoba, A.; Armendariz, A. M.; Teran Toledo,
X.; Romo Garcia, J.; Fernandez, H.; Rozhon, E. J.; Antiviral
Res. 1997, 35, 91. 167. Singer, C.; Underwood, E. A.; A Short History of Medicine,
Oxford University Press: New York, 1962, p. 674. 168. Van Duuren, B. L.; Orris, L., Arroyo, E.; Nature 1963, 200, 1115. 169. Arroyo, E. R.; Holcomb, J.; J. Acknowledgments M.; Santos, C. F.; Braz. J. Med. Biol. Res. 2002, 35,
1215. 136. Barbosa, P. R.; Fascio, M.; Martins, D.; Roque, N. F.; Arkivoc
2004, 6, 95. 137. Milo, B.; Risco, E.; Vila, R.; Iglesias, J.; Canigueral, S.; C.; J. Nat. Prod. 2002, 65, 1143. 159. Suárez, A. I.; Blanco, Z.; Compagnone, R. S.; Salazar-Bookaman,
M. M.; Zapata, V.; Alvarado, C.; J. Ethnopharmacol. 2006, 105,
99. 138. Barry, P.E.; Hipp, A.L.; Wurdack, K.J.; Van Ee, B.; Riina, R.;
Am. J. Bot. 2005, 92, 1520. 160. Santos, F. A.; Jeferson, F. A.; Santos, C. C.; Silveira, E. R.;
Rao, V. S. N.; Life Sci. 2005, 77, 2953. 139. Gershenzon, J.; J. Chem. Ecol. 1994, 20, 1281. Traditional uses, Chemistry and Pharmacology of Croton species S.; Durán, N.; Haun, M.; Toxicology
2003, 188, 261. 178. Anazetti, M. C.; Melo, P. S.; Durán, N.; Haun, M.; Toxicology
2004, 203, 123. 200. Melo, P. S.; Durán, N.; Hiruma-Lima, C. A.; Souza-Brito, A. R. M.; Haun, M.; J. Ethnopharmacol. 2003, 87, 169. 201. Ushiyama, S.; Matsuda, K.; Asai, F.; Yamazaki, M.; Biochem. Pharmacol. 1987, 36, 369. 179. Agner, A. R.; Maciel, M. A. M.; Pinto, A. C.; Pamplona, S. G. S. R.; Colus, I. M. S.; Teratogenesis Carcinog. Mutagen. 1999,
19, 377. 202. Takagi, A.; Koga, W.; Aiba, Y.; Kabir, A. M.; Watanabe, S.;
Ohta-Tada, U.; Osaki, T.; Kamiya, S.; Miwa, T.; J. Gastroenterol. Hepatol. 2000, 15, 374. 180. Agner, A. R.; Maciel, M. A. M.; Pinto, A. C.; Colus, I. M. S.;
Planta Med. 2001, 67, 815. 203. Koga, T.; Inoue, H.; Ishii, C.; Okazaki, Y.; Domon, H.; Utsui,
Y.; J. Antimicrob. Chemother. 2002, 50, 133. 181. Correa, D. H. A.; Melo, P. S.; de Carvalho, C. A. A.; de Azevedo,
M. B. M.; Durán, N.; Haun, M.; Eur. J. Pharmacol. 2005, 510, 17. 182. Block, S.; Gerkens, P.; Peulen, O.; Jolois, O.; Mingeot-Leclercq,
M. P.; De Pauw-Gillet, M. C.; Quetin-Leclerq, J.; Anticancer
Res. 2005, 25, 363. 204. Magalhães, P. J. C.; Lahlou, S.; Leal-Cardoso, J. H.; Fundam. Clin. Pharmacol. 2004, 18, 539. 205. Albuquerque, A. A. C.; Sorenson, A. L.; Leal-Cardoso, J. H.;
J. Ethnopharmacol. 1995, 49, 41. 183. Cragg, G. M.; Newman, D. J.; Ann. Appl. Biol. 2003, 143, 127. 184. Kupchan, S. M.; Uchida, I.; Branfman, A. R.; Dailey, R. G.;
Fel, B. Y.;.Science 1976, 191, 571. 206. Di Cesare, D.; DuPont, H. L.; Mathewson, J. J.; Ashley, D.;
Martinez-Sandoval, F.; Pennington, J. E.; Porter, S. B.; Am. J. Gastroenterol. 2002, 97, 2585. 185. Kawai, K.; Tsuno, N. H.; Kitayama, J.; Okaji, Y.; Yazawa, K.;
Asakage, M.; Yamashita, H.; Watanabe, T.; Takahashi, K.;
Nagawa, H.; Anticancer Drugs 2005, 16, 401. 207. Adam, G.; Ousingsawat, J.; Schreiber, R.; Kunzelmann, K.;
Pflugers Arch. 2005, 449, 470. Vol. 18, No. 1, 2007 33 Salatino et al. 208. Holodniy, M.; Koch, J.; Mistal, M.; Am. J. Gastroenterol. 1999,
94, 3267. 213. Norte, M. C. B.; Cosentino, R. M.; Lazarini, C. A.;
Phytomedicine 2005, 12, 294. 209. Bighetti, E. J. B.; Hiruma-Lima, C. A.; Gracioso, J. S.; Brito, 214. Costa, A. M. L.; Silva, J. C. R.; Campos, A. R.; Rao, V. S. N.;
Maciel, M. A. Traditional uses, Chemistry and Pharmacology of Croton species M.; Pinto, A. C.; Phytother. Res. 1999, 13, 689. A. R. M.; J. Pharm. Pharmacol. 1999, 51, 1447. Maciel, M. A. M.; Pinto, A. C.; Phytother. Res. 1999, 13, 689. 215. Smitt, O.; Hogberg, H-E.; Tetrahedron 2002, 58, 7691. 210. Ichiara, Y.; Takeya, K.; Hitotsuyanagi, Y.; Morita, H.; Okuyama,
S.; Suganuma, M.; Fujiki, H.; Motidome, M.; Itokawa, H.;
Planta Med. 1992, 58, 549. 216. Fairnsworth, N. R. In Biodiversity; Wilson, E. O.; Peter, F. M., eds., National Academy Press: Washington D.C., 1988,
p. 83. Planta Med. 1992, 58, 549. 211. Carvalho, J. C. T.; Silva, M. F. C.; Maciel, M. A. M.; Pinto, A. C.; Nunes D. S.; Mendes Lima, R.; Bastos, J. K.; Sarti, S. J.;
Planta Med. 1996, 62, 402. 217. Rudall, P.; Ann. Mo. Bot. Gard.1994, 81, 270. 218. Fahn, A.; New Phytol. 1988, 108, 229. 218. Fahn, A.; New Phytol. 1988, 108, 229. Planta Med. 1996, 62, 402. 212. Silva, R. M.; Oliveira, F. A.; Cunha, K. M. A.; Maia, J. L.;
Maciel, M. A. M.; Pinto, A. C.; Nascimento, N. R. F.; Santos,
F. A.; Rao, V. S. N.; Vasc. Pharmacol. 2005, 43, 11. Received: August 9, 2006
Web Release Date: January 26, 2007 F. A.; Rao, V. S. N.; Vasc. Pharmacol. 2005, 43, 11. FAPESP helped in meeting the publication costs of this article.
|
https://openalex.org/W4288098351
|
https://link.springer.com/content/pdf/10.1007/978-3-030-22974-0.pdf
|
English
| null |
The 14th International Congress on Applied Mineralogy (ICAM-2019)
|
DOAJ (DOAJ: Directory of Open Access Journals)
| 2,019
|
cc-by
| 188,307
|
Springer Proceedings in Earth and Environmental Sciences
14th International
Congress for
Applied Mineralogy
(ICAM2019)
Belgorod State Technological University
named after V. G. Shukhov,
23–27 September 2019,
B l
d R
Sergey Glagolev Editor Springer Proceedings in Earth and Environmental Sciences 14th International
Congress for
Applied Mineralogy
(ICAM2019) Belgorod State Technological University
named after V. G. Shukhov,
23–27 September 2019,
Belgorod, Russia Belgorod State Technological University
named after V. G. Shukhov,
23–27 September 2019, p
Belgorod, Russia Springer Proceedings in Earth
and Environmental Sciences Series Editor
Natalia S. Bezaeva, The Moscow Area, Russia Series Editor The series Springer Proceedings in Earth and Environmental Sciences publishes
proceedings from scholarly meetings and workshops on all topics related to
Environmental and Earth Sciences and related sciences. This series constitutes a
comprehensive up-to-date source of reference on a field or subfield of relevance in
Earth and Environmental Sciences. In addition to an overall evaluation of the
interest, scientific quality, and timeliness of each proposal at the hands of the
publisher, individual contributions are all refereed to the high quality standards of
leading journals in the field. Thus, this series provides the research community with
well-edited, authoritative reports on developments in the most exciting areas of
environmental sciences, earth sciences and related fields. More information about this series at http://www.springer.com/series/16067 Sergey Glagolev
Editor Editor
Sergey Glagolev
Belgorod State Technological University
Belgorod, Russia ISSN 2524-342X
ISSN 2524-3438
(electronic)
Springer Proceedings in Earth and Environmental Sciences
ISBN 978-3-030-22973-3
ISBN 978-3-030-22974-0
(eBook)
https://doi.org/10.1007/978-3-030-22974-0 This Springer imprint is published by the registered company Springer Nature Switzerland AG
The registered company address is: Gewerbestrasse 11, 6330 Cham, Switzerland ISSN 2524-342X
ISSN 2524-3438
(electronic)
Springer Proceedings in Earth and Environmental Sciences
ISBN 978-3-030-22973-3
ISBN 978-3-030-22974-0
(eBook)
https://doi.org/10.1007/978-3-030-22974-0 © The Editor(s) (if applicable) and The Author(s) 2019. This book is an open access publication. © The Editor(s) (if applicable) and The Author(s) 2019. This book is an open access publication. Open Access This book is licensed under the terms of the Creative Commons Attribution 4.0 Open Access This book is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing, adap-
tation, distribution and reproduction in any medium or format, as long as you give appropriate credit to
the original author(s) and the source, provide a link to the Creative Commons license and indicate if
changes were made. The images or other third party material in this book are included in the book’s Creative Commons
license, unless indicated otherwise in a credit line to the material. If material is not included in the book’s
Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the
permitted use, you will need to obtain permission directly from the copyright holder. p
y
p
y
py g
The use of general descriptive names, registered names, trademarks, service marks, etc. in this
publication does not imply, even in the absence of a specific statement, that such names are exempt from
the relevant protective laws and regulations and therefore free for general use. The publisher, the authors and the editors are safe to assume that the advice and information in this
book are believed to be true and accurate at the date of publication. Neither the publisher nor the
authors or the editors give a warranty, expressed or implied, with respect to the material contained
herein or for any errors or omissions that may have been made. The publisher remains neutral with regard
to jurisdictional claims in published maps and institutional affiliations. This Springer imprint is published by the registered company Springer Nature Switzerland AG
The registered company address is: Gewerbestrasse 11, 6330 Cham, Switzerland Foreword Dear Participants of the 14th International Congress for Applied Mineralogy! On behalf of the Presidium of the Russian Mineralogical Society, let us con-
gratulate you with this important scientific event and express our hope for the
successful and productive work at its sessions and symposia. Challenges that will be considered at the Congress, which is held in Russia for
the first time, are exceptionally broad. They coincide with many directions that
were recently developed by the Russian Mineralogical Society at its Commissions
on technological mineralogy, environmental mineralogy and geochemistry, crystal
chemistry and spectroscopy, and organic mineralogy. In 2017, the Russian Mineralogical Society (RMS) celebrated its 200th
anniversary, and today, it unites more than twelve hundred members from different
fields and disciplines. RMS sections are present in many scientifically important
cities and mining regions of our country. RMS members are actively solving
important problems of development and production of raw materials for nuclear,
diamond, rare metal, and oil industries. The continuing progress in microscopy (electron, tunnel, atomic force) brought
mineralogical studies to the micro- and nanolevels. Intervention into micro- and
nanoworlds is an important new stage of development of modern mineralogy and
mineralogical crystallography. Due to the success in these fields, a range of new
possibilities was opened for: prospecting and elaboration of novel types of mineral deposits, prospecting and elaboration of novel types of mineral deposits, solution of the remediation problems for mining regions and the rational use of
mine tailings,
deeper understanding of the nature of oil fields, organic minerals and
biominerals,
creation of principally new geo-inspired materials with improved functional
properties. solution of the remediation problems for mining regions and the rational use of
mine tailings, solution of the remediation problems for mining regions and the rational use of
mine tailings,
deeper understanding of the nature of oil fields, organic minerals and
biominerals,
creation of principally new geo inspired materials with improved functional deeper understanding of the nature of oil fields, organic minerals and
biominerals, creation of principally new geo-inspired materials with improved functional
properties. v v vi Foreword Foreword The methods of mineralogical sciences are still in the focus of problems that are
faced by Earth sciences, rational use of natural resources, conservation of natural
diversity, and cultural heritage of the humankind. Foreword The methods of mineralogical sciences are still in the focus of problems that are
faced by Earth sciences, rational use of natural resources, conservation of natural
diversity, and cultural heritage of the humankind. We are sure that the Congress will demonstrate a broad spectrum of important
scientific discoveries, will develop new directions for research and exploration in
applied mineralogy, and, last but not least, will provide a lavish basis for new
professional contacts and collaborations. Wishing you great inspiration, new discoveries, and achievements. Yury B. Marin
President of the Russian Mineralogical Society,
Corresponding Member of the Russian Academy of Sciences Sergey V. Krivovichev
Vice-President of the Russian Mineralogical Society,
Corresponding Member of the Russian Academy of Sciences Sergey V. Krivovichev
Vice-President of the Russian Mineralogical Society,
Corresponding Member of the Russian Academy of Sciences Sergey N. Glagolev
Editor
Rector of Belgorod State Technological University named after V. G. Shukhov,
Doctor of Economics, Professor,
Co-chairman of the National Organizing Committee Preface Dear Participants of the Congress! I welcome you to Belgorod at the 14th International Congress for Applied
Mineralogy ICAM 2019! Considering a special significance and prestige of the events of the International
Council on Applied Mineralogy and the key role of our country as one of the world
leaders in the industrial sectors associated with mineralogy, it is gratifying to note
that the Congress is held on the base of Belgorod Technological University, which
is a flagship university of Russia, an educational and scientific-innovative center of
attraction for talents and formation of a professional regional elite, the personnel
base and potential of enterprises of mining and processing industries. Main scientific trends of the forthcoming Congress are devoted to modern
methods of studying rocks and minerals, geometallurgy and processing, biomin-
eralogy, functional materials, hydrocarbon raw materials, and environmental
problems of the industry. Undoubtedly, this large-scale and productive meeting of
budding and leading specialists of the industry will be an important milestone in the
history of applied mineralogy, will determine promising areas of industrial devel-
opment of this science in Russia and abroad, and will contribute to overcoming
significant technological barriers, creating new and improving existing industry
collaborations. Colleagues, I wish you successful reports and presentations, constructive dia-
logue, recognition of colleagues and like-minded people, strengthening the
deserved authority of domestic science in the global educational and professional
community! Sergey N. Glagolev
Editor
Rector of Belgorod State Technological University named after V. G. Shukhov,
Doctor of Economics, Professor,
Co-chairman of the National Organizing Committee vii National Scientific Committee Leonid Vaisberg (Chairman, Academician RAS)
Nikolay Bortnikov (Co-chairman, Academician RAS)
Olga Kotova (Executive Secretary)
Askhab Askhabov (Academician RAS)
Yury Marin (Corresponding Member RAS)
Nikolay Sobolev (Academician RAS)
Tamara Matveeva
Valentin Chanturiya (Academician RAS)
Elena Ozhogina
Sergey Krivovichev (Corresponding Member RAS)
Vladimir Shchiptsov
Olga Frank-Kamenetskaya (Associate Member) ix 14th ICAM x x Supporting from IMA-Commission for Applied Mineralogy Maarten A. T. M. Broekmans (Chairman IMA-CAM), Norway
Jan Elsen (Secretary IMA-CAM), Belgium ICAM Council Saverio Fiore (President), Italy
Olga Kotova (Vice-President), Russia
Dieter Rammlmair (Secretary), Germany International Board of Reviewers xi 14th ICAM xi 14th ICAM Organizers
General Sponsor
Information Support 14th ICAM Organizers 14th ICAM Organizers 14th ICAM Organizers
General Sponsor
Information Support General Sponsor General Sponsor Information Support Information Support Information Support Information Support Contents Geometallurgy, Technological Mineralogy and Processing
of Mineral Raw
Scherrer Width and Topography of Illite as Potential Indicators
for Contrasting Cu-Recovery by Flotation of a Chilean Porphyry
Cu (Mo) Ore . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
G. Abarzúa, L. Gutiérrez, U. Kelm, and J. Morales
Ore Mineralogical Study of Cerattepe Au-Cu (–Zn) VMS Deposit
(Artvin-Turkey) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
İ. Akpınar and E. Çiftçi
Correlation Value of the Mineralogical Composition of Tills
in the North of European Russia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
L. Andreicheva
Comparative Gold Deportment Study on Direct Leaching and Hybrid
Process Tails of Oxide Ores from Mayskoye Au Deposit
(Chukotka, Russia) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
I. Anisimov, A. Dolotova, A. Sagitova, M. Kharitonova, B. Milman,
and I. Agapov
Mineral-Geochemical Criteria to Gold and Silver Recovery
for Geometallurgical Sampling Campaign on Primorskoe
Gold-Silver Deposit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
I. Anisimov, A. Sagitova, O. Troshina, and I. Agapov
Mineralogical-Geochemical Criteria for Geometallurgical Mapping
of Levoberezhnoye Au Deposit (Khabarovsk Region, Russia) . . . . . . . . 24
I. Anisimov, A. Sagitova, M. Kharitonova, A. Dolotova, and I. Agapov Geometallurgy, Technological Mineralogy and Processing
of Mineral Raw xiii Contents xiv Mineralogical Reasons of Au Recovery Variability from
North-Western Pit of Varvara Au-Cu Mine (Kazakhstan)
and Criteria for Geometallurgical Mapping . . . . . . . . Contents . . . . . . . . . . . . . . 29
I. Anisimov, A. Dolotova, A. Sagitova, M. Kharitonova, and I. Agapov
Mineralogical Breakthrough into Nanoworld: Results and Challenges . . . 33
A. Askhabov
Case Study: Geochemistry and Mineralogy of Copper Mine Tailings
in Northern Central-Chile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
K. Berkh, D. Rammlmair, M. Drobe, and J. Meima
Application of Fluoride Technology for Processing of Off-Grade
Aluminum Raw Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
I. Burtsev, I. Perovskiy, and D. Kuzmin
Applied Mineralogy for Complex and Profound Mineral Processing . . . 45
V. Chanturiya and T. Matveeva
Mineralogical and Technological Features of Tin Minerals
at Pravourmiysky Deposit (Khabarovsk Region) . . . . . . . . . . . . . . . . . . 49
T. Chikisheva, S. Prokopyev, E. Kolesov, V. Kilin, A. Karpova,
E. Prokopyev, and V. Tukuser
Ore Mineralogy of High Sulfidation Çorak-Taç Epimesothermal
Gold Deposit (Yusufeli-Artvin-Turkey) . . . . . . . . . . . . . . . . . . . . . . . . . 53
K. Diarra, E. Sangu, and E. Çiftçi
Mineralogical and Technological Aspects of Phosphate
Ore Processing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
A. Elbendari, V. Potemkin, T. Aleksandrova, and N. Nikolaeva
Properties and Processing of Ores Containing Layered Silicates . . . . . . 66
A. Gerasimov, V. Arsentyev, and V. Lazareva
Applied Mineralogy of Anthropogenic Accessory Minerals . . . . . . . . . . 70
A. Gerasimov, E. Kotova, and I. Ustinov
Crystal-Chemical and Technological Features of the KMA
Natural Magnetites . . . . . . . . . . . . . . . . . . . . . . . . . . . Contents . . . . . . . . . . . . . . 75
T. Gzogyan and S. Gzogyan
Practical Application of Technological Mineralogy on the Example
of Studying of Sulphidization in the KMA Ferruginous Quartzites . . . . 80
S. Gzogyan and T. Gzogyan
Ore Mineralogy of Kirazliyayla (Yenişehir-Bursa-Turkey)
Mesothermal Zn-Pb-(–Cu) Deposit: Preliminary Results . . . . . . . . . . . . 84
F. Javid and E. Çiftçi Contents xv Use of Borogypsum as Secondary Raw . . . . . . . . . . . . . . . . . . . . . . . . . 90
A. Khatkova, L. Nikitina, and S. Pateyuk
New Approaches in X-ray Phase Analysis of Gypsum Raw Material
of Diverse Genesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
V. Klimenko, V. Pavlenko, and T. Klimenko
Nanotechnologies in Mineral-Geochemical Methods for Assessing
the Forms of Finding of Gold, Related Elements, Technological
Properties of Industrial Ores and Their Tails . . . . . . . . . . . . . . . . . . . . 99
R. Koneev, R. Khalmatov, O. Tursunkulov, A. Krivosheeva,
N. Iskandarov, and A. Sigida
Applied Mineralogy of Mining Industrial Wastes. . . . . . . . . . . . . . . . . . 103
O. Kotova and E. Ozhogina
High-Tech Elements in Minerals of Massive Sulfide Deposits:
LA-ICP-MS Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107
V. Maslennikov, S. Maslennikova, N. Aupova, A. Tseluyko, R. Large,
L. Danyushevsky, and U. Yatimov
Absolutely Pure Gold with High Fineness 1000‰ . . . . . . . . . . . . . . . . . 111
Z. Nikiforova
New Data on Microhardness of Placer Gold . . . . . . . . . . . . . . . . . . . . . 115
Z. Contents Nikiforova
Modern Methods of Technological Mineralogy in Assessing
the Quality of Rare Metal Raw Materials . . . . . . . . . . . . . . . . . . . . . . . 119
E. Ozhogina, A. Rogozhin, O. Yakushina, Yu. Astakhova,
E. Likhnikevich, N. Sycheva, A. Iospa, and V. Zhukova
Topochemical Transformations in Sodium-Bismuth-Silicate System
at 100–900 °C . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
A. Pavlenko and R. Yastrebinskiy
Ag-Bearing Mineralization of Nevenrekan Deposit
(Magadan Region, Russia) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
E. Podolian, I. Shelukhina, and I. Kotova
Th/U Relations as an Indicator of the Genesis of Metamorphic
Zircons (On the Example of the North of the Urals) . . . . . . . . . . . . . . . 129
Y. Pystina and A. Pystin
Phenomenon of Microphase Heterogenization by Means
of Endocrypt-Scattered Impurity of Rare and Noble Metals
as a Result of Radiation by Accelerated Electrons of Bauxites . . . . . . . . 133
I. Razmyslov, O. Kotova, V. Silaev, and L. A. Gomze xvi Gold Extraction to Ferrosilicium, Production of Foam Silicate from
Processing Tails of the Olimpiada Mining and Processing Complex
Gold Processing Plant (Russia, Krasnoyarsk Territory) . . . . . . . . . . . . . 136
A. Sazonov, V. Pavlov, S. Silyanov, and E. Zvyagina
Predictive Assessment of Quality of Mineral
Aggregates Disintegration. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140
S. Shevchenko, R. Brodskaya, I. Bilskaya, Yu. Kobzeva,
and V. Lyahnitskaya
Development of Methods for Anti-filtration Formations
Destruction Inside a Heap Leach Pile. . . . . . . . . . . . . . . . . . . . . . . . . . . 143
H. Tcharo, M. Koulibaly, and F. K. N. Contents Tchibozo
Microtomographic Study of Gabbro-Diabase Structural
Transformations Under Compressive Loads . . . . . . . . . . . . . . . . . . . . . 146
L. Vaisberg and E. Kameneva
Process Mineralogy as a Basis of Molybdoscheelite Ore Preparation . . . 152
L. Vaisberg, O. Kononov, and I. Ustinov
Crystallomorphology of Cassiterite and Its Practical Importance . . . . . 157
I. Vdovina
Modal Analysis of Rocks and Ores in Thin Sections . . . . . . . . . . . . . . . 162
Yu. Voytekhovsky
Quality Assurance Support (QA/QC System)
of Mineralogical Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
O. Yakushina, E. Gorbatova, E. Ozhogina, and A. Rogozhin
Mineral Preparation in Geological Research . . . . . . . . . . . . . . . . . . . . . 172
T. Yusupov, A. Travin, S. Novikova, and D. Yudin
Industrial Minerals, Precious Stones, Ores and Mining
Impact Diamonds: Types, Properties and Uses . . . . . . . . . . . . . . . . . . . 179
V. Afanasiev, N. Pokhilenko, A. Eliseev, S. Gromilov, S. Ugapieva,
and V. Senyut
Authentic Semi-precious and Precious Gemstones of Turkey:
Special Emphasis on the Ones Preferred for Prayer Beads . . . . . . . . . . 183
E. Çiftçi, H. Selim, and H. Sendir
Biooxidation of Copper Sulfide Minerals . . . . . . . . . . . . . . . . . . . . . . . . 189
Yu. Elkina, E. Melnikova, V. Melamud, and A. Bulaev
Genetic Problem of Quartz in Titanium Minerals in Paleoplacers
of Middle Timan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192
I. Golubeva, I. Burtsev, A. Ponaryadov, and A. Shmakova xvii Contents Gold and Platinum Group Minerals (PGM) from the Placers
of Northwest Kuznetsk Alatau (NWKA) (South Siberia, Russia) . . . . . . 195
V. Contents Gusev, S. Zhmodik, G. Nesterenko, and D. Belyanin
Noble Metal Mineralization of the PGM Zone “C” of the East-Pana
Layered Intrusion (Kola Peninsula) . . . . . . . . . . . . . . . . . . . . . . . . . . . . 198
O. Kazanov, G. Logovskaya, and S. Korneev
Shungites and Their Industrial Potential . . . . . . . . . . . . . . . . . . . . . . . . 201
V. Kovalevski and V. Shchiptsov
Gold-Silver Natural Alloy of Chromitites from the Kamenushinsky
Massif (The Middle Urals) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205
A. Minibaev
Microbial Processes in Ore-Bearing Laterite at the Tomtor Nb-REE
Deposit: Evidence from Carbon Isotope Composition in Carbonates . . . 208
V. Ponomarchuk, E. Lazareva, S. Zhmodik, N. Karmanov, and A. Piryaev
Peridot: Types of Deposits and Formation Conditions . . . . . . . . . . . . . . 212
S. Sokolov
Mineralogical Analysis of Glacial Deposits and Titanium
Paleoplacers of the East European Part of Russia . . . . . . . . . . . . . . . . . 214
N. Vorobyov and A. Shmakova
Oil and Gas Reservoirs, Including Gas Hydrates
A Bench Scale Investigation of Pump-Ejector System
at Simultaneous Water and Gas Injection . . . . . . . . . . . . . . . . . . . . . . . 219
S. Karabaev, N. Olmaskhanov, N. Mirsamiev, and J. Mugisho
Integrated Use of Oil and Salt Layers at Oil Field Development . . . . . . 221
V. Malyukov and K. Vorobyev
Oil and Gas Reservoirs in the Lower Triassic Deposits
in the Arctic Regions of the Timan-Pechora Province . . . . . . . . . . . . . . 223
N. Timonina
Associated Petroleum Gas Flaring: The Problem
and Possible Solution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
A. Contents Vorobev and E. Shchesnyak
Innovative Technology of Using Anti-sand Filters at Wells
of the Vankor Oil and Gas Field . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 231
K. Vorobyev and A. Gomes xviii Contents Analytical Methods, Instrumentation and Automation
Thermometry of Apatite Saturation (The Kozhym Massif,
The Subpolar Urals) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 235
Y. Denisova, A. Vikhot, O. Grakova, and N. Uljasheva
Studies of Structural Changes in Surface and Deep Layers
in Magnetite Crystals After High Pressure Pressing. . . . . . . . . . . . . . . . 239
P. Matyukhin
The Potential of Lacquer Peel Profiles and Hyperspectral Analysis
for Exploration of Tailings Deposits . . . . . . . . . . . . . . . . . . . . . . . . . . . . 244
W. Nikonow and D. Rammlmair
Methods of Extraction of Micro- and Nanoparticles of Metal
Compounds from Fine Fractions of Rocks, Ores
and Processing Products. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 248
A. Smetannikov and D. Onosov
Advanced Materials with Improved Characteristics, Including
Technical Ceramics and Glass
Efficiency Evaluation for Titanium Dioxide-Based Advanced
Materials in Water Treatment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 255
M. Harja, O. Kotova, S. Sun, A. Ponaryadov, and T. Shchemelinina
The Use of Karelia’s High-Mg Rocks for the Production of Building
Materials, Ceramics and Other Materials with Improved Properties . . . 259
V. Ilyina
Kinetic Features of Formation of Supramolecular Matrices
on the Basis of Silica Monodisperse Spherical Particles . . . . . . . . . . . . . 263
D. Kamashev
Three-Cation Scandium Borates RxLa1−XSc3(BO3)4(R = Sm, Tb):
Synthesis, Structure, Crystal Growth and Luminescent Properties . Contents . . . 267
A. Kokh, A. Kuznetsov, K. Kokh, N. Kononova, V. Shevchenko,
B. Uralbekov, A. Bolatov, and V. Svetlichnyi
Rational Usage of Amorphous Varieties of Silicon Dioxide in Dry
Mixtures of Glass with Specific Light Transmittance . . . . . . . . . . . . . . . 272
N. Min’ko and O. Dobrinskaya
Peculiarities of Phase Formation in Artificial Ceramic Binders
for White-Ware Compositions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 277
I. Moreva, E. Evtushenko, O. Sysa, and V. Bedina Analytical Methods, Instrumentation and Automation Contents xix Experimental Modeling of Biogeosorbents . . . . . . . . . . . . . . . . . . . . . . . 281
T. Shchemelinina, O. Kotova, E. Anchugova, D. Shushkov,
G. Ignatyev, and M. Markarova
Heating Rate and Liquid Glass Content Influence on Cement
Brick Dehydration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 286
V. Strokova and D. Bondarenko
Structure and Surface Reactivity Mediated Enzymatic
Performances of Clay-Based Nanobiocatalyst . . . . . . . . . . . . . . . . . . . . . 290
S. Sun, K. Wang, F. Dong, B. Ma, T. Huo, Y. Zhao, H. Yu, Y. Huang,
and J. Huang
Structural-Phase Stabilization of Clay Materials
in Hydrothermal Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 292
O. Sysa, E. Evtushenko, I. Moreva, and V. Loktionov
Phase Changes in Radiation Protection Composite Materials Based
on Bismuth Oxide. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 296
S. Yashkina, V. Doroganov, E. Evtushenko, O. Gavshina, and E. Sysa
Development of Technology for Anti-corrosion Glass Enamel
Coatings for Oil Pipelines . . . . Contents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 300
E. Yatsenko, A. Ryabova, and L. Klimova
Building Materials
Optimization of Formulations of Cement Composites Modified
by Calcined Clay Raw Material for Energy Efficient Building
Constructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 307
A. Balykov, T. Nizina, V. Volodin, and D. Korovkin
Santa Maria Clays as Ceramic Raw Materials . . . . . . . . . . . . . . . . . . . . 311
Â. Cerqueira, C. Sequeira, D. Terroso, S. Moutinho, C. Costa,
and F. Rocha
Alkaline Activation of Rammed Earth Material – “New Generation
of Adobes” . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 313
C. Costa, D. Arduin, C. Sequeira, D. Terroso, S. Moutinho, Â. Cerqueira,
A. Velosa, and F. Rocha
Structurization of Composites When Using 3D-Additive Technologies
in Construction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 315
M. Elistratkin, V. Lesovik, N. Chernysheva, E. Glagolev, and P. Hardaev
Influence of Flow Blowing Agent on the Properties of Aerated
Concrete Variable Density and Strength . . . . . . . . . . . . . . . . . . . . . . . . 319
V. Galdina, E. Gurova, P. Deryabin, M. Rashchupkina, and I. Chulkova Optimization of Formulations of Cement Composites Modified
by Calcined Clay Raw Material for Energy Efficient Building
Constructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Contents . . . . . . . . 307
A. Balykov, T. Nizina, V. Volodin, and D. Korovkin
Santa Maria Clays as Ceramic Raw Materials . . . . . . . . . . . . . . . . . . . . 311
Â. Cerqueira, C. Sequeira, D. Terroso, S. Moutinho, C. Costa,
and F. Rocha
Alkaline Activation of Rammed Earth Material – “New Generation
of Adobes” . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 313
C. Costa, D. Arduin, C. Sequeira, D. Terroso, S. Moutinho, Â. Cerqueira,
A. Velosa, and F. Rocha
Structurization of Composites When Using 3D-Additive Technologies
in Construction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 315
M. Elistratkin, V. Lesovik, N. Chernysheva, E. Glagolev, and P. Hardaev
Influence of Flow Blowing Agent on the Properties of Aerated
Concrete Variable Density and Strength . . . . . . . . . . . . . . . . . . . . . . . . 319
V. Galdina, E. Gurova, P. Deryabin, M. Rashchupkina, and I. Chulkova Contents xx Structuring Features of Mixed Cements on the Basis
of Technogenic Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 323
M. Garkavi, A. Artamonov, E. Kolodezhnaya, A. Pursheva,
and M. Akhmetzyanova
Use of Slags in the Production of Portland Cement Clinker. . . . . . . . . . 327
V. Konovalov, A. Fedorov, and A. Goncharov
Geopolymerization and Structure Formation in Alkali Activated
Aluminosilicates with Different Crystallinity Degree. . . . . . . . . . . . . . . . 331
N. Kozhukhova, V. Strokova, I. Zhernovsky, and K. Sobolev
Matrix Instruments for Calculating Costs of Concrete
with Multicomponent Binders . . . . . . . . . . . . . . . Contents . . . . . . . . . . . . . . . . . 335
T. Kuladzhi, S.-A. Murtazaev, S. Aliev, and M. Hubaev
Characterisation of Perovskites in a Calcium Sulfo Aluminate Cement. . . 339
G. Le Saout, R. Idir, and J.-C. Roux
Geonics (Geomimetics) as a Theoretical Basis for New
Generation Compositing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 344
V. Lesovik, A. Volodchenko, E. Glagolev, I. Lashina, and H.-B. Fischer
Regularities in the Formation of the Structure and Properties
of Coatings Based on Silicate Paint Sol . . . . . . . . . . . . . . . . . . . . . . . . . 348
V. Loganina, E. Mazhitov, and V. Demyanova
Influence of Sodium Oxide on Brightness Coefficient
of Portland Cement Clinker . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 352
D. Mishin and S. Kovalyov
Production of Bleached Cement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 356
D. Mishin and S. Kovalev
Multicomponent Binders with Off-Grade Fillers . . . . . . . . . . . . . . . . . . 360
S.-A. Murtazaev, M. Salamanova, M. Saydumov, A. Alaskhanov,
and M. Khubaev
High-Quality Concretes for Foundations of the Multifunctional
High-Rise Complex (MHC) «Akhmat Tower» . . . . . . . . . . . . . . . . . . . . 365
S.-A. Murtazaev, M. Saydumov, A. Alaskhanov, and M. Nakhaev
Designing High-Strength Concrete Using Products of Dismantling
of Buildings and Structures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 369
T. Murtazaeva, A. Alaskhanov, M. Saidumov, and V. Contents Hadisov
Estimation of Rheo-Technological Effectiveness of Polycarboxylate
Superplasticizer in Filled Cement Systems in the Development
of Self-compacting Concrete for High-Density Reinforced
Building Constructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 372
T Ni i
A B l k
V V l di
d D K
ki Contents xxi Parameters of Siliciferous Substrate of Photocatalytic Composition
Material as a Factor of Its Efficiency . . . . . . . . . . . . . . . . . . . . . . . . . . . 376
Y. Ogurtsova, E. Gubareva, M. Labuzova, and V. Strokova
Properties Improvement of Metakaolin-Zeolite-Diatomite-Red
Mud Based Geopolymers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 381
F. Rocha, C. Costa, W. Hajjaji, S. Andrejkovičová, S. Moutinho,
and A. Cerqueira
Features of Production of Fine Concretes Based on Clinkerless
Binders of Alkaline Mixing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 385
M. Salamanova, S.-A. Murtazaev, A. Alashanov, and Z. Ismailova
Impact of Thermal Modification on Properties of Basalt Fiber
for Concrete Reinforcement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389
V. Strokova, V. Nelyubova, I. Zhernovsky, O. Masanin, S. Usikov,
and V. Babaev
Activation of Cement in a Jet Mill . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 393
S. Titov and A. Kazakov
The Law of Similarity and Designing High-Performance Composites . . . 395
A. Tolstoy, V. Lesovik, E. Glagolev, and L. Zagorodniuk
Genesis of Clay Rock of the Incomplete Stage of Mineral Formation
as a Raw Material Base for Autoclaved Materials . . . . . . . . . . . . . . Contents . . . 399
A. Volodchenko and V. Strokova
Abnormal Mineral Formation in Aluminate Cement Stone . . . . . . . . . . 403
I. Zhernovsky, V. Strokova, V. Nelyubova, Yu. Ogurtsova,
and M. Rykunova
Structural Transformations of Low-Temperature Quartz
During Mechanoactivation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 407
I. Zhernovsky and V. Strokova
Biomimetic Materials on a Mineral Basis, Biomineralogy
Effect of Earthquake on the Landscape of Jiuzhaigou-Huanglong
Travertine and Its Restoration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 413
F. Dong, Q. Dai, Q. Li, F. Wang, and Y. Luo
Microbial Colonies in Renal Stones . . . . . . . . . . . . . . . . . . . . . . . . . . . . 415
A. Izatulina, M. Zelenskaya, and O. Frank-Kamenetskaya
Fabrication of ZnO/Palygorskite Nanocomposites
for Antibacterial Application . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 419
Y. Kang, A. Hui, and A. Wang xxii Bacterial Oxidation of Pyrite Surface . . . . . . . . . . . . . . . . . . . . . . . . . . . 423
S. Lipko, I. Lipko, K. Arsent’ev, and V. Tauson
Biomimetic Superhydrophobic Cobalt Blue/Clay Mineral Hybrid
Pigments with Self-cleaning Property and Different Colors . . . . . . . . . . 427
B. Mu, A. Zhang, and A. Wang
Silicon Dioxide in Mineralized Heart Valves . . . . . . . . . . . . . . . . . . . . . 432
A. Titov, V. Zaikovskii, and P. M. Larionov
Preparation of Macroporous Adsorbent Based on Montmorillonite
Stabilized Pickering Medium Internal Phase Emulsions . . . . . . . . . . . . . 436
F. Wang, Y. Zhu, W. Wang, and A. Wang
Environment and Energy Resources
Depletion of the Land Resources and Its Effect on the Environment . Contents . . 443
M. Abou Zahr Diaz, M. A. Alawiyeh, and M. Ghaboura
Geochemical Behavior of Heavy Metals During Treatment by
Phosphoric Fertilizer at a Dumping Site in Kabwe, Zambia . . . . . . . . . 445
H. Kamegamori, K. Lawrence, T. Sato, and T. Otake
Murataite-Pyrochlore Ceramics as Complex Matrices for Radioactive
Waste Immobilization: Structural and Microstructural Mechanisms
of Crystallization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 447
S. Krivovichev, S. Yudintsev, A. Pakhomova, and S. Stefanovsky
Cs Leaching Behavior During Alteration Process of Calcium Silicate
Hydrate and Potassium Alumino Silicate Hydrate . . . . . . . . . . . . . . . . . 451
K. Kuroda, K. Toda, Y. Kobayashi, T. Sato, and T. Otake
Environmental Pollution Problems in the Mining Regions of Russia . . . 453
E. Levchenko, I. Spiridonov, and D. Klyucharev
Environmental Solutions for the Disposal of Fine White
Marble Waste . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 457
I. Shadrunova, T. Chekushina, and A. Proshlyakov
Security Test of New Technology in View of Increased Performance
of Oil Platforms Without Increasing Environmental Risks. . . . . . . . . . . 461
E. M. Tanoh Boguy and T. Chekushina
Calcite Mineral Generation in Cold-Water Travertine
Huanglong, China . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 463
F. Wang, F. Dong, X. Zhao, Q. Dai, Q. Li, Y. Luo, and S. Deng Contents Optimization of the Natural-Technical System “Iron Ore Quarry”
Management Based on the Algorithm of the Rock Mass
Stability Ensuring. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Contents . . . . 466
L. Yarg, I. Fomenko, and D. Gorobtsov
Utilization of Associated Oil Gas: Geo-ecological Problems
and Modernization of the State . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 471
L. Z. Zhang and H. Y. Sun
Cultural Heritage, Artifacts and Their Preservation
Identifying the Decorative Stone Samples from the Mining
Museum’s Collection: First Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . 475
N. Borovkova and M. Machevariani
Monitoring of the State of St. Petersburg Stone Monuments
and the Strategy of Their Preservation . . . . . . . . . . . . . . . . . . . . . . . . . 479
O. Frank-Kamenetskaya, D. Vlasov, V. Rytikova, V. Parfenov,
V. Manurtdinova, and M. Zelenskaya
Ceramics Sugar Jars Pieces from Aveiro Production . . . . . . . . . . . . . . . 483
S. Moutinho, C. Costa, Â. Cerqueira, C. Sequeira, D. Terroso, J. Nobre,
P. Morgado, A. Velosa, and F. Rocha
Author Index. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 485 Scherrer Width and Topography of Illite
as Potential Indicators for Contrasting Cu-
Recovery by Flotation of a Chilean Porphyry
Cu (Mo) Ore G. Abarzúa1, L. Gutiérrez2, U. Kelm1(&), and J. Morales3 1 Instituto de Geología Económica Aplicada (GEA),
Universidad de Concepción, Concepción, Chile
ukelm@udec.cl, ued.kelm@gmail.com
2 Departamento de Ingeniería Metalúrgica, Facultad de Ingeniería,
Universidad de Concepción, Concepción, Chile
3 Departamento de Geología, Universidad de Salamanca, Salamanca, Spain 1 Instituto de Geología Económica Aplicada (GEA),
Universidad de Concepción, Concepción, Chile
ukelm@udec.cl, ued.kelm@gmail.com 1 Instituto de Geología Económica Aplicada (GEA),
Universidad de Concepción, Concepción, Chile
ukelm@udec.cl, ued.kelm@gmail.com
2 Departamento de Ingeniería Metalúrgica, Facultad de Ingeniería,
Universidad de Concepción, Concepción, Chile
3 Departamento de Geología, Universidad de Salamanca, Salamanca, Spain 2 Departamento de Ingeniería Metalúrgica, Facultad de Ingeniería Universidad de Concepción, Concepción, Chile 3 Departamento de Geología, Universidad de Salamanca, Salamanca, Spai 3 Departamento de Geología, Universidad de Salamanca, Salamanca, Spain Abstract. Two contrasting feeds in terms of copper recovery from a Cu
(Mo) porphyry deposit but with similar overall mineralogy have been charac-
terized by X-ray diffraction for their <1 µm fraction illite crystallinity, Scherrer
width and by atomic force microscopy for surface roughness. The unfavorable
feed displayed slightly higher crystallinity, larger Scherrer width and surface
roughness factors, than the feed with good Cu recovery. As Scherrer width is an
easy and cheaply to determine parameter it is suggested as complementary
information to particle size distribution analyses when dealing with feeds where
illite may affect pulp viscosity or gangue adhesion to bubbles during flotation. Keywords: Illite Scherrer width Surface roughness Flotation Sericite
Atomic force microscopy © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 3–6, 2019.
https://doi.org/10.1007/978-3-030-22974-0_1 G. Abarzúa et al. Cheng and Peng (2018) suggest negative effects for low crystallinity kaolinite rich
ore, much of the work being based on artificial ore-gangue mixtures. However Jorjani
et al. (2011) single out illite and vermiculate as key gangue affecting flotation for the
Iranian Sarcheshmeh porphyry copper deposit. The present exploratory study has been
sparked by the effort to develop a formula for blending ore based on mineralogical-
chemical parameters of a giant porphyry copper deposit in Chile. Ore with similar
sericitic alteration and chalcopyrite dominated ore phases, were identified by the
concentrator operation as favorable (F) and unfavorable (UF) floating feed, the latter
entering the concentrator only as blend. Given this overall mineralogical similarity, it
was decided to characterize the contrasting feeds based on their illite crystallinity –
Scherrer width and concomitant surface roughness. 2
Methods and Approaches Triplicate samples of favorable (F) and unfavorable (UF) floating ore were prepared for
X-ray diffraction (XRD) analysis of the clay size fraction (>0.45 <1 µm) based on the
recommendations of Moore and Reynolds (1997). XRD measurements were carried out
on a Bruker D4 diffractometer operated with Ni-filtered Cu-radiation. Illite crystallinity
and Scherrer width were determined on the 001 basal reflection following Lorentzian
adjustment using the Origin 8.5 program. Atomic Force Microscope (AFM) measure-
ments of topography were carried out with an AIST-NT equipment in contact mode on
a 5 5 µm surface. WSxM5.0 software was used for calculating Ra (arithmetic
average) and Rrms (root mean square) roughness factors (Horcas et al. 2007; Erinosho
et al. 2018). 1
Introduction For over half a century, illite crystallinity has been used as an indicator of mineral
maturity in metasediments between the transition of diagenesis to very low temperature
metamorphism and the incipient low-grade metamorphism or epizone (Frey 1999). Illite crystallinity (later Kübler Index) measured at full width half medium height
(FWHM) of the basal XRD-reflection is also an indirect indicator for the size of the
jointly diffracting illite sheets, also known as Scherrer width, which has been directly
visualized with the widespread availability of Transmission Electron Microscopy (Frey
1999). Superimposition of metamorphic and hydrothermal alteration processes, paired
with time consuming analytical routines, has limited the application to alteration halos
of ore deposits; an exception is the study by Beaufort et al. (2005) on the East Alligator
River Uranium deposit in the Northern Territory, Australia, due to its abundance of
chlorite and illite gangue. Sericitic alteration (muscovite/illite) also represents wide-
spread gangue for Andean Cu (Mo) porphyry ore deposits, but systematic studies of
phyllosilicate crystallinity or Scherrer width within ore deposit areas are not available. 4 G. Abarzúa et al. 3
Results and Discussion Illite crystallinity values of both samples (F: 0.15 D° 2H, UF: 0.12 D° 2H) correspond
to epizone values, and for the unfavorable feed are at the sensitivity limit of this method
(Frey 1999). Though values for the favorable feed are marginally lower, nevertheless
this difference is expressed in an increased Scherrer width or crystallite size (F:
48.8 nm, UF: 62.6 nm) for the unfavorable feed. Topographic images of sample surfaces show different roughness, being the
favorable feed (F) the smoother. Roughness factors were calculated for different sur-
faces scanned by AFM. As a mean, 10 sample surfaces were measured and statistically
compared, giving values of Ra = 57 ± 20 and Rrms = 66 ± 23 for the favorable feed
and Ra = 68 ± 18 and Rrms = 86 ± 19 for the unfavourable feed sample. Undoubt-
edly, different scales of images analysed imply changes in the surface parameters. To
avoid this, images with the same size have been compared. As it can be observed in the
Fig. 1, roughness parameters confirm the XRD results. Despite the difficulties of differentiating between illite generations in rocks with
sericitic overprint in porphyry (and other) ore deposits, this simple XRD measurement
permits concomitant calculation of coherently diffracting particle sizes or Scherrer
width for a given geometallurgical unit. The correspondence observed for this Scherrer Width and Topography of Illite as Potential Indicators Scherrer Width and Topography of Illite as Potential Indicators 5 Fig. 1. Topographic aspects of F-1 sample (left) and UF-1 sample (right) Fig. 1. Topographic aspects of F-1 sample (left) and UF-1 sample (right) exploratory study between increased Scherrer width and higher AFM measured surface
roughness, recommends the XRD based value as an easily available tool to assess
difference in flotation for samples where traditional methods like optical microscopy,
automated mineralogy, and semi-quantitative whole rock mineralogy do not reveal
obvious mineralogical differences between feeds with contrasting flotation behavior. Farrokhpay and Ndlovu (2013) discussed the effect of clay particle size on pulp rhe-
ology; here the X-ray coherent Scherrer width is suggested as a complementary indi-
cator to the particle size distribution measurements by laser diffraction in the clay size
range. The scale of surface roughness as a factor impacting on particle adhesion to
bubbles has been studied by Nikolaev (2016). 3
Results and Discussion However, direct AFM measurements are
still no routine procedures to define geometallurgical units within an ore deposit,
whereas XRD information can be generated faster and in a more standardized fashion. 4
Conclusions Illite crystallinity, Scherrer width and AFM-determined surface roughness have been
determined for two flotation-feed of a Cu (Mo) porphyry copper deposit with con-
trasting Cu-recovery. Samples did not display any mineralogical difference allowing a
straightforward explanation of this difference. For this exploratory study case, Scherrer
width is considered an easy to obtain parameter that points to differences in surface G. Abarzúa et al. 6 roughness of illite particles in the <1 µm fraction and thus may influence pulp viscosity
and/or particle adhesion to bubbles during flotation. roughness of illite particles in the <1 µm fraction and thus may influence pulp viscosity
and/or particle adhesion to bubbles during flotation. Acknowledgements. Dr Manuel Melendrez, Departamento de Ingeniería de Materiales,
Universidad de Concepción is thanked for access to the AFM equipment. Acknowledgements. Dr Manuel Melendrez, Departamento de Ingeniería de Materiales,
Universidad de Concepción is thanked for access to the AFM equipment. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 7–10, 2019.
https://doi.org/10.1007/978-3-030-22974-0_2 References Beaufort D, Patrier P, Laverret E (2005) Clay alteration associated with proterozoic
unconformity-type uranium deposit in the East Alligator Rivers uranium fields, Northern
Territory, Australia. Econ Geol 100:515–536 Chen X, Peng Y (2018) Managing clay minerals in froth flotation. A critical review. Miner
Process Extr Metall Rev 39(5):289–307 Erinosho MF, Akinlabi ET, Johnson OT (2018) Characterization of surface roughness of laser
deposited titanium alloy and copper using AFM. Appl Surf Sci 435:393–397 Farrokhpay S, Ndlovu B (2013) Effect of phyllosilicate minerals on the rheology, colloidal and
flotation behaviour of chalcopyrite mineral. In: Australasian conference on chemical
engineering, Chemeca 2013, Challenging Tomorrow, p 733 Frey M (1999) Very low-grade metamorphism of clastic sedimentary rocks: in Low Temperature
Metamorphism. Blackie and Sons, Glasgow, pp 9–58 Horcas I, Fernández R, Gomez-Rodriguez JM, Colchero JWSX, Gómez-Herrero JWSXM,
Baro AM (2007) WSXM: a software for scanning probe microscopy and a tool for
nanotechnology. Rev Sci Instrum 78:013705 Jorjani E, Barkhordari HR, Khorami MT, Fazeli A (2011) Effects of aluminosilicate minerals on
copper–molybdenum flotation from Sarcheshmeh porphyry ores. Miner Eng 24(8):754–759
Moore DM, Reynolds RC Jr (1997) X-ray diffraction and the identification and analysis of clay Jorjani E, Barkhordari HR, Khorami MT, Fazeli A (2011) Effects of aluminosilicate minerals on
copper–molybdenum flotation from Sarcheshmeh porphyry ores. Miner Eng 24(8):754–759 Moore DM, Reynolds RC Jr (1997) X-ray diffraction and the identification and analysis of clay
minerals. Oxford University Press, Oxford Nikolaev A (2016) Flotation kinetic model with respect to particle heterogeneity and roughness. Int J Miner Process 155:74–82 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction The eastern Pontide orogenic belt of Turkey is an important segment of the Tethyan-
Eurasian Metallogenic Belt. This belt carries a special importance in metallogeny of
Turkey and hosts numerous VMS deposit. (Akıncı 1984; Çiftçi and Hagni 2005;
Güven 1993; Revan et al. 2013; Zaykov et al. 2006; Yiğit 2005). The most of the
eastern Pontide VMS deposits show some similarities in many aspects to the Kuroko
deposits of Japan (Çiftçi and Hagni 2005; Ciftci 2000; Pejatoviç 1979). The Cerattepe
Deposit is a Kuroko-type VMS deposit located in late Cretaceous age volcanic,
intrusive and sedimentary rocks. It is distinguished by an unusual basal zone of high-
grade copper sulfides and an overlying Au-rich oxide zone from the other VMS
deposits of northeastern Pontides. Ore Mineralogical Study of Cerattepe Au-Cu
(–Zn) VMS Deposit (Artvin-Turkey) İ. Akpınar1(&) and E. Çiftçi2 1 Department of Geological Engineering, Faculty of Engineering and Natural Science,
Gumushane University, Gumushane, Turkey
hiakpinar@gumushane.edu.tr
2 Department of Geology, Faculty of Mines, Istanbul Technical University,
Istanbul, Turkey Abstract. The Cerattepe mine, one of the volcanogenic massive sulfide
deposits in northeastern Turkey, is hosted within the late Cretaceous volcanic,
intrusive and sedimentary rocks. Deposit’s main ore body contains high-grade
massive copper sulfides and a gold-silver and barium rich oxide zone, charac-
terized by dense alteration stages, is situated on top of it. Replacement, cata-
clastic, breccia, dissemination, dendritic, concentric growth, colloform, and
framboidal textures were identified. Pyrite, sphalerite, marcasite, chalcopyrite,
bornite, galena, tennantite-tetrahedrite, gold, covellite, digenite, chalcocite,
cuprite and cubanite constitute the mineral paragenesis where quartz, calcite and
barite account for the gangue minerals. Limonite, hematite, lepidocrocite,
malachite, azurite and jarosite developed in the oxidation zone. Keywords: Eastern Pontide Cerattepe VMS deposit Framboidal pyrite
Bird’s eye-texture Keywords: Eastern Pontide Cerattepe VMS deposit Framboidal pyrite
Bird’s eye-texture Keywords: Eastern Pontide Cerattepe VMS deposit Framboidal pyrite
Bird’s eye-texture 2
Methods and Approaches A total of 46 samples representing oxide (14), sulfide (28) and stringer zones (4) of
Cerattepe ore deposit were collected from drill cores, underground audits and surface
outcrops. Polished sections were prepared and Nikon Eclipse LV100 reflected light İ. Akpınar and E. Çiftçi 8 microscopy was employed for examination. The ore minerals and the paragenesis were
identified on the basis of their petrographical features and their textural relationships,
respectively. Electron Probe Micro Analysis (EPMA) and Secondary Electron
Microscopy-Energy Dispersive Spectroscopy (SEM-EDS) were used for chemistry of
sulfide minerals. 3
Results and Discussion The mineral paragenesis (Fig. 1) of Cerattepe VMS deposit comprises of pyrite,
sphalerite, marcasite, chalcopyrite, bornite-idaite, galena, covellite, chalcocite, cuban-
ite, cuprite including sulfosalts (mainly tennantite and lesser tetrahedrite) gold, silver,
arsenopyrite and bournonite. Fig. 1. Ore mineral paragenetic sequence of Cerattepe Au-Cu (±Zn) VMS deposit Fig. 1. Ore mineral paragenetic sequence of Cerattepe Au-Cu (±Zn) VMS deposit Gangue minerals include barite, quartz, gypsum, anhydrite and calcite. Hematite,
limonite, lepidocrocite, malachite and azurite, and jarosite are the oxidation minerals. Four generation of pyrite is specified. Pyrite I is represented by colloform-concentric
textured (Fig. 2) grains formed from initial solution reached the seafloor from the
chimney. The colloform textured pyrites are progressively overgrown later coarse
crystalline grains, which are the second generation pyrites. These are later extensively altered to marcasite. The framboidal-pelletal textured
collomorphic grains, seen sometimes coeval with sphalerite and galena, are the third
generation pyrites. The fourth generation pyrites are small sized euhedral, subhedral
grains observed on transition zone of sulfide ore- footwall rock and veinlets of stringer
zone. Ore Mineralogical Study of Cerattepe Au-Cu (±Zn) VMS Deposit 9 py2
py2
cv
cp
a
sl1
py1
b
py1
cp
py3
py3
py3
py2
gn + td
py2
mrc
sl1
ba
sl1
py1
py + mrc
py +mrc
py
cv
bn
py + mrc
py
sl1
py
bn
bn
cv
cv
cp
Py
ba
py2
sl2
c
b
f
d
e
g
h
cp
Fig. 2. Photomicrographs of selected polished sections showing observed textures in Cerattepe
deposit. (a) Cataclastic texture of pyrite grains. (b) Colloform textured pyrite and marcasite
intergrowth surrounded by sphalerite “Bird’s eye texture”; (c) Colloform textured pyrite
aggregate with a spongy textured intergrowth zones comprised of sphalerite, galena and
tetrahedrite; (d) Three different generation of pyrites, (e) Pelletal pyrite framboids with minor
sphalerite and galena in the cracks; (f) Bornite and chalcopyrite replaced by covellite;
(g) collomorphically banded sulfides in melnikovite pyrite; (h) Dissociation texture of
chalcopyrite in sphalerite replaced by marcasite py2
py2
cv
cp
a
sl1
py1
b
py1
cp
py3
py3
py3
py2
gn + td
py2
mrc
sl1
ba
sl1
py1
py + mrc
py +mrc
py
cv
bn
py + mrc
py
sl1
py
bn
bn
cv
cv
cp
Py
ba
py2
sl2
c
b
f
d
e
g
h
cp b d e h Fig. 2. Photomicrographs of selected polished sections showing observed textures in Cerattepe
deposit. 3
Results and Discussion (a) Cataclastic texture of pyrite grains. (b) Colloform textured pyrite and marcasite
intergrowth surrounded by sphalerite “Bird’s eye texture”; (c) Colloform textured pyrite
aggregate with a spongy textured intergrowth zones comprised of sphalerite, galena and
tetrahedrite; (d) Three different generation of pyrites, (e) Pelletal pyrite framboids with minor
sphalerite and galena in the cracks; (f) Bornite and chalcopyrite replaced by covellite;
(g) collomorphically banded sulfides in melnikovite pyrite; (h) Dissociation texture of
chalcopyrite in sphalerite replaced by marcasite Most of the minerals are small - fine grained, and the larger grains of the major
minerals are in the order of 100–800 µm up to 1.2–2 mm in size and some rare pyrite
grains have a size of 5 mm. Most of the minor and trace minerals are much smaller,
typically in the order of 1–20 µm in size. Majority of ore minerals are anhedral with the
exception of pyrite, quartz, barite and some sphalerite and galena occur as euhedral to
subhedral crystals. Observed ore textures are dissemination and veinlet textures in the
stockwork and siliceous ore zones, whereas replacement, overgrowth, concentric and
colloform textures become prevalent in the massive ore, particularly in the center of
main deposit. Banded textures of black and yellow ore are seen with polymetallic sulfides. In the
outer part of the main ore body, at lateral zones, clastic or fragmental ore textures are
present. İ. Akpınar and E. Çiftçi İ. Akpınar and E. Çiftçi 10 4
Conclusions It is concluded that the ore mineral assemblage and textures observed in Cerat-
tepe VMS deposit are comparable to those of other VMS deposits occur in north
eastern Pontide region, Kuroko deposits of Japan and also comparable modern sea-
floor–seamount VMS deposits in the world. Acknowledgements. This study was financially supported by BAP Project unit (No: 39615) of
Istanbul Technical University in Turkey. We are thankful to Mr. Ünsal Arkadaş and staff of
Etibakır A.Ş. in Artvin, Turkey for their courteous support for the field work. References Akıncı ÖT (1984) The geology and the metallogeny of the Eastern Pontides (Turkey). In:
International world geological congress abstracts, pp 197–198 Ciftci E (2000) Mineralogy, paragenetic sequence, geochemistry, and genesis of the gold and
silver bearing upper cretaceous mineral deposits, Northeastern Turkey. Ph.D. Thesis,
University of Missouri-Rolla, Rolla, MO, USA (unpublished) y
p
Çiftçi E, Hagni RD (2005) Mineralogy of the Lahanos deposit a Kuroko-type VMS deposit from
the eastern Pontides (Giresun, NE Turkey). Geol Bull Turk 48(1):55–64 üven İH (1993) Artvin Kafkasör Sahası Maden Jeolojisi Raporu No 2600, MTA, Ankara
metallogeny. Ore Geol Rev 147–179 g
y
Pejatovic S (1979) Metallogeny of the Pontid type massive sulphide deposits, Ankara, 98 p Revan MK, Genç Y, Maslennikov V, Ünlü T, Delibaş O, Hamzaçebi S (2013) Original findings
on the ore-bearing facies of volcanogenic massive sulphide deposits in the eastern black sea
region (NE Turkey). Bull MTA 147:73–89 g
y
Yigit O (2005) Gold in Turkey - a missing link and metallogenic features of the in Tethyan igit O (2005) Gold in Turkey - a missing link and metallogenic features of the in Tethyan Yigit O (2005) Gold in Turkey - a missing link and metallogenic features of the in Tethyan
Zaykov V, Novoselov K, Kotlyarov V (2006) Native gold and tellurides in the Murgul and
Çayeli volcanogenic Cu deposits (Turkey) Zaykov V, Novoselov K, Kotlyarov V (2006) Native gold and tellurides in the Murgul and
Çayeli volcanogenic Cu deposits (Turkey) Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction The studies were conducted on a vast and geologically heterogeneous territory of the
Timan-Pechora-Vychegda region, which determined the variability of lithological and
mineralogical parameters of the tills in term of the regional plan. This restricts their use
for correlations. But taking into account the factors of the glacial sedimentation genesis
and the complex study of the tills, the stratigraphic confinement of moraine horizons is
established very confidently. The features of the mineral-petrographic composition of
the tills and the rocks of the source provinces (remote, transit and local) are discussed
in detail in the papers by Andreicheva (1994, 2017). Correlation Value of the Mineralogical
Composition of Tills in the North
of European Russia L. Andreicheva(&)
Institute of Geology KomiSC UB RAS, Syktyvkar, Russia
andreicheva@geo.komisc.ru L. Andreicheva(&) Institute of Geology KomiSC UB RAS, Syktyvkar, Russia
andreicheva@geo.komisc.ru Abstract. The material composition of the tills, including its mineral compo-
nent, is formed during of the exaration-accumulative activity of the cover glacier
and depends on the composition of the rocks of the glaciation centers, transit
areas, local underlying rocks and on the relief of the preglacial bed. Thus the
composition of the till is conditioned by the total influence of source glacier
provinces and represents an average sample of the rocks on the way of the
glacier. Keywords: Mineral composition Correlation Till
Petrographic composition © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 11–14, 2019.
https://doi.org/10.1007/978-3-030-22974-0_3 2
Results and Discussion Pomusovski (Okskii) till in the lower Pechora and Laya is characterized by a low
content of a heavy fraction (0.26–0.4%) and amphibole (10–12%) - garnet (12–19%) -
epidote (35–41%) mineral association. The concentration of titanium minerals (rutile,
titanite, leucoxene) is 7–9%, the total amount of pyrite and siderite does not exceed 10–
11%. Eastward (in the valleys of the Kolvaand Pechora Rvers - in borehole 301-
Kushshor) the yield of the heavy fraction is significantly higher - 0.8–1.16%, but the
epidote content decreases (to 26–30%), amphiboles (to 5–7%) and the total concen-
tration of pyrite and siderite increases sharply to 37%. In borehole 301-Kushshor the © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 11–14, 2019. https://doi.org/10.1007/978-3-030-22974-0_3 L. Andreicheva 12 number of garnets (up to 23%) and the content of titanium minerals (up to 11%)
increase. The decrease in the content of amphiboles in the Pomusovskitill eastward with the
underlying Aptian and Albianrocks. They are characterized by the epidote-amphibole
mineral association with a distinct W-E decreasing content of amphiboles. The
enrichment of the heavy fraction with pyrite and siderite in the of Kolvavalley is
associated with assimilation of these minerals from the Mesozoic sediments. The
petrographic composition of the fragments and their south-eastward orientation testifies
to the formation of the pomusovskytill in the region due to source from Fennoscan-
dinavia and the Northern Timan. The composition of the heavy fraction of Pechora(Dneprovian) till is also char-
acterized by areal variability, reflecting the specifics of the mineral composition of the
rocks of remote, transit and local source provinces. The minimum yield of the heavy
fraction is 0.33%, noted in Pechoratill in the middle Pechora River, the maximum -
0.86%, in the SeydaRver. The mineral association is represented by epidote, garnets,
amphiboles, pyrite, siderite and ilmenite. In some areas, the role of titanium minerals
increases, most often due to an increase in the amount of leucoxene, sometimes the
content of metamorphogenic minerals is increased: kyanite, staurolite, sillimanite. Epidote prevails in the Pechoratill, its quantity varies from 14 to 37%. Maximum
concentrations are recorded in sections of the middle Pechora, the lowest - in the
ChernayaRver. The number of garnets is increased to 21% in the ShapkinaRver,
reduced to 5% - on the valley of the SeydaRver. 2
Results and Discussion The content of amphiboles is low: its
maximum concentration (16%) is marked in the VychegdaRver, and the lowest - on the
right bank of the UsaRver, where is only 0.6%, which is associated with the complete
absence of amphiboles in the underlying Mesozoic rocks. The amount of pyrite also
varies significantly: its minimum content (3%) is in the basin of the ShapkinaRver, the
maximum in the SeydaRver- 27%. Siderite in the LayaRver is 8%, in the sections in the
ChernayaRver reaches of 27%, pyrite and siderite are constantly present. Their total
concentrations are different, but the ratio is stable: almost everywhere siderite domi-
nates pyrite. In the petrographic thin sections of the pechorskytill, glauconite is con-
tained in significant quantities (up to 60 grains in a standard petrographic section),
which, like pyrite, siderite, is characteristic of local Mesozoic rocks. The composition
of the minerals of the heavy fraction testifies to the participation of underlying Triassic,
Jurassic and Cretaceous rocks in the formation of the mineral spectrum of the till
(Andreicheva and Nikitenko 1989). Single reference boulders are constantly present -
pink crinoid-bryozoan Novaya Zemlya limestones. Another characteristic feature of the
Pechora glaciation is a steady south-western trend of ice movement, which is consistent
with the petrographic composition of rock fragments in the till. The obtained data
testify to the Paykhoy-Ural-Novaya Zemlya center of glaciation in the Pechora time
throughout the region. The peculiarity of the paleogeography of Vychegda (Moskovian) glaciation is the
existence in the west and in the east of the region of the ice covers of various centers. Accordingly, the material composition of the till was formed by the material of Correlation Value of the Mineralogical Composition 13 different source provinces. In the western and central parts of the region, the heavy
fraction of till, which ranges from 0.42% in the lower Pechora River to 0.95% in the
Vychegda valley, is represented by an amphibole-garnet-epidote mineral association
with increased total pyrite and siderite contents (up to 32% in the ChernayaRver valley
in the north of the Bolshezemelskaya tundra). Sometimes the amounts of ilmenite and
titanium minerals are increased. In the south of the region, the association of heavy
minerals contains less epidote than in the Pechoratill, and there are more amphiboles
and garnets, sometimes amphiboles reach 57% of the heavy fraction. 2
Results and Discussion Pyrite and siderite
make up the first percentages of the weight of the heavy fraction, and ilmenite con-
centrations are increased (9-12%). The amount of glauconite in Vychegdatill is 3–4
times lower than in Pechoratill: up to 15–20 grains per standard petrographic section. Heteroagedtills particularly sharply differ from one another by the composition of
heavy minerals in the southern regions of the region. The orientation of the fragments
in sector 270–3600 and the presence of indicator rocks of the North-West Terrigenous
Mineralogical Province also indicate the migration of material from the west and from
the north-west. Another feature of this till is the predominance of light colored lime-
stones in the group of carbonate rocks. In the northeast of the region, the mineral composition of the heavy fraction of the
Vychegdatill is variable and consists of ilmenite (12%), garnet (15%), pyrite (15%),
siderite (25%), epidote (32%). The content of titanium minerals is slightly increased –
10%, and amphiboles – is decreased (4–12%). The petrographic spectrum of fragments
represents Uralian rocks. The fragments of rocks are oriented in sub latitudinal and SW
directions, i.e. material for the formation of Vychegdatill came from the east-northeast. The main mineral association of the Polar (Ostashkovian) till contains epidote (19–
27%), garnets (14–20%) and amphibole (11–16%). The total content of siderite and
pyrite is 13–35% with dominating of siderite. The crinoid-bryozoanNovaya Zemlya
limestones are present. The orientation of detrital material from NNE to SSW confirms
the onset of the cover glacier in polar time from Pay-Khoy-Novaya Zemlya. Acknowledgements. This research was supported by UB RAS project №18-5-5-50 and project
AAAA-A17-117121140081-7. 3
Conclusions Despite the variability of lithologic and mineralogical parameters of tills in terms of the
regional plan, the data of the mineral composition of tills can be used with confidence
for the dismemberment of Quaternary sections and wide spatial correlations of glacial
deposits, taking into account the factors of glacial sedimentation genesis, as well as the
petrographic composition of rock fragments and their orientation. L. Andreicheva 14 Andreicheva LN (2017) Correlation of neopleistocene tills of the north of the Russian plain in
petrographic composition. Lithol Miner (1):82–94. (in Russian) Andreicheva LN (1994) The source provinces and their influence on the formation of the
composition of the moraines of the Timan-Pechora-Vychegda region. Lithol Miner (1):127–
131. (in Russian) Andreicheva LN, Nikitenko IP (1989) The mineral composition of the fine earths of the main
moraines of the Timan-Pechora-Vychegda region. In: Mineralogy of the Timan-Northern Ural
region, Syktyvkar, proceedings of the institute of geology, Komi SC UB RAS, no 72, pp 52–
62. (in Russian) Andreicheva LN (2017) Correlation of neopleistocene tills of the north of the Russian plain in
petrographic composition. Lithol Miner (1):82–94. (in Russian)
Andreicheva LN (1994) The source provinces and their influence on the formation of the Andreicheva LN (2017) Correlation of neopleistocene tills of the north of the Russian plain in
petrographic composition. Lithol Miner (1):82–94. (in Russian)
Andreicheva LN (1994) The source provinces and their influence on the formation of the
composition of the moraines of the Timan-Pechora-Vychegda region. Lithol Miner (1):127–
131. (in Russian)
Andreicheva LN, Nikitenko IP (1989) The mineral composition of the fine earths of the main
moraines of the Timan-Pechora-Vychegda region. In: Mineralogy of the Timan-Northern Ural
region, Syktyvkar, proceedings of the institute of geology, Komi SC UB RAS, no 72, pp 52–
62. (in Russian) References Andreicheva LN (2017) Correlation of neopleistocene tills of the north of the Russian plain in
petrographic composition. Lithol Miner (1):82–94. (in Russian) Andreicheva LN (1994) The source provinces and their influence on the formation of the
composition of the moraines of the Timan-Pechora-Vychegda region. Lithol Miner (1):127–
131. (in Russian) Andreicheva LN, Nikitenko IP (1989) The mineral composition of the fine earths of the main
moraines of the Timan-Pechora-Vychegda region. In: Mineralogy of the Timan-Northern Ural
region, Syktyvkar, proceedings of the institute of geology, Komi SC UB RAS, no 72, pp 52–
62. (in Russian) Andreicheva LN, Nikitenko IP (1989) The mineral composition of the fine earths of the main
moraines of the Timan-Pechora-Vychegda region. In: Mineralogy of the Timan-Northern Ural
region, Syktyvkar, proceedings of the institute of geology, Komi SC UB RAS, no 72, pp 52–
62. (in Russian) Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. I. Anisimov(&), A. Dolotova, A. Sagitova, M. Kharitonova,
B. Milman, and I. Agapov I. Anisimov(&), A. Dolotova, A. Sagitova, M. Kharitonova,
B. Milman, and I. Agapov Science and Technology Research Division, Polymetal Engineering,
Saint-Petersburg, Russia
anisimovis@polymetal.ru Science and Technology Research Division, Polymetal Engineering,
Saint-Petersburg, Russia
anisimovis@polymetal.ru Abstract. Mayskoye gold deposit is located in Chukotka, Russia. Ore bodies
are mineralized brecciation zones composed of vein-quartz, argillic altered rocks
(siltstone and carbonaceous silts) with fine disseminated and veinlet gold-
bearing arsenopyrite and pyrite. Two main technological types of ores were
distinguished at the deposit: primary and oxide. The main reserves were rep-
resented by primary ores, which are classified as refractory. Oxide ore had a
quartz-micaceous composition with minor feldspars, kaolinite and sulfides. Beside native visible and colloidal gold, other main carriers of gold in oxide ore
are arsenopyrite, pyrite, minerals of scorodite group and stibnite. Tested oxide
ore sample showed low recoveries, according to the existing flow sheet in the
CIL plant. Cyanidation tests showed some preg-robbing effect on organic
matter. Flotation of carbonatious matter with consecutive leaching of flotation
tails proved to have better recoveries than direct leaching by reducing preg-
robbing on carbon three times. Keywords: Refractory gold Preg-robbing Invisible gold
Surface contamination © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 15–18, 2019.
https://doi.org/10.1007/978-3-030-22974-0_4 I. Anisimov et al. I. Anisimov et al. The studied oxidized ore sample had low gold recovery in CIL plant, thus directed
to gold deportment analysis and lab test work. The sample mineral composition was
quartz-micaceous with minor kaolinite and secondary minerals scorodite, jarosite, tri-
puhyite and others. Natural coal content was 1,4%. Sulfides were represented by pyrite,
marcasite, arsenopyrite and stibnite. Oxidized ore appeared to be partially refractory. Most of the refractory gold was represented by “invisible” gold. It was contained both
in the native form, and was dispersed in liberated ore minerals: pyrite, arsenopyrite,
stibnite and scorodite, and their binaries with gangue. The form of occurrence of gold
in the ore and its connection with mineral carrier was important to determine effective
methods for metal extraction. Such study in plant products was necessary to chase the
issue and find optimization in reagent scheme or the flow-sheet. 2
Samples and Methods of Study Two samples of initial oxidized ores from 1st and 2nd ore zones of Mayskoe gold
deposit and four plant products: direct cyanidation tails, flotation tails and combined
leach tails of flotation tails leaching product. The examination methodology included
the following stages: classification of samples into narrow size classes, separation of
material in heavy liquids with density of 2,90 g/cm3 and 2,5 g/cm3. Gold concentrate
was panned from the heavy concentrate. The following separation products were
obtained: gold gravity concentrate, heavy concentrate, rock-forming lights and car-
bonaceous fraction. Bulk mineral composition of the initial size classes and lights was
studied by powder X-ray phase analysis using the Rietveld refinement. The mineral
composition of the gold concentrates and heavy concentrates were studied by optical
microscopy and the mineral identification of the carbonaceous fraction was done with
stereo microscope. Gold content in the products was analyzed by fire assays; arsenic, antimony and
other elements by XRF in all products, except for gold concentrates. In the gold tips,
the quantitative finding of gold was carried out by optical and electron microscopy. Chemical composition of minerals-carrier of gold and other ore minerals was
studied with SEM-EDX. 1
Introduction Mayskoye Au deposit is located in Chaunsky region in Chukotka, Russia. The ore was
formed by quartz-pyrite-arsenopyrite shear opening veins in terrigenous sequence of
carbonaceous
siltstone
and
sandy-siltstones. Ore
bearing
sequence
altered
to
micaceous-carbonate-albite rocks (of beresite formation). Thus, the ore had quartz-
micaceous composition with minor feldspars and sulphide material. Major part of gold
was refractory and concentrated in arsenopyrite and partially pyrite. Thus primary ores
are treated with flotation recovering around 90% of gold with sulfide concentrate,
which was shipped to Amursky POX plant for oxidation and cyanide tank leaching. Part of the carbonaceous matter is preg-robbing gold. Upper part of the deposit
contains oxide ore, which mostly was not refractory and free leaching. The carbon in
pulp plant treats oxide ore during the warm season. 16 3
Results and Discussion Quartz and phyllosilicates (muscovite, illite, smectite, kaolinite, dickite) were the main
minerals in the samples; scorodite, jarosite, sulfides and other minerals were acces-
sories. The main minerals-carrier of gold are represented by arsenopyrite, pyrite,
marcasite and stibnite, oxidized forms of arsenic - scorodite, and antimony - tripuhyite,
stibiconite, cervantite and valentinite. Native carbon content in the ore was 0,1–1,4%. Visible native gold in samples had high fineness (902–914‰). The distribution
pattern of visible native gold particles in ore zone 1 and 2 were slightly different. 70%
of the visible native gold from head sample from the first ore zone was larger than
90 µm. 65% of the visible native gold from the second ore zone head sample was
coarser than 70 µm. All visible gold particles in the cyanidation tails of first ore zone Comparative Gold Deportment Study on Direct Leaching and Hybrid Process 17 were less than 45 µm. Gold particles coarser than 45 µm dominated in cyanidation tails
of the sample from the second ore zone. Native gold observed in the samples of direct
leach tails were found in free particles with the surface shielded or in binaries with
oxidized antimony forms. Rarely binaries of gold with pyrite, stibnite and gangue
minerals were observed. Fineness of visible gold in cyanidation tails was 10–15%
lower than in initial products with higher silver content. Besides natural gold the metal
was in dispersed form in the mineral-carriers: pyrite, arsenopyrite, stibnite, scorodite,
antimony oxidized forms and bound with carbon (Fig. 1). Fig. 1. Gold deportment in the plant feed, direct leach tailings, flotation tails and flotation tails
leach residues of oxidized ores from 1st and 2nd ore zones of Mayskoe gold deposit Fig. 1. Gold deportment in the plant feed, direct leach tailings, flotation tails and flotation tails
leach residues of oxidized ores from 1st and 2nd ore zones of Mayskoe gold deposit Free leaching gold accounted for 49% in head sample from the first ore zone and
64% in head sample from the second ore zone. The rest of the gold was on carbon
(about 22–26%), in sulfides (about 5–10%) and scorodite (about 9–15%). Visible gold
extraction by direct cyanidation was in range of 96–99% and gold from scorodite only
53–61%. 3
Results and Discussion The losses of visible gold in leaching occurred due to the blocking of the
surface with compounds of oxidized antimony, as well as the slower cyanidation of
gold with lower fineness. Coal flotation reduced gold losses by 22%. 4
Conclusions Gold in the head samples was represented by both visible high-grade gold, most of
which was a of gravity size and invisible/colloidal gold (<0,5 micron), forming more
than half of the metal of the original ore. Lower grade gold particles leached slower. p
p
y
g
g
g
,
which was a of gravity size and invisible/colloidal gold (<0,5 micron), forming more
than half of the metal of the original ore. Lower grade gold particles leached slower. The oxide ore of zones 1 and 2 of Mayskoye deposit was partially refractory, since
most of the gold was invisible and dispersed in arsenopyrite, scorodite and with carbon. The oxide ore of zones 1 and 2 of Mayskoye deposit was partially refractory, since
most of the gold was invisible and dispersed in arsenopyrite, scorodite and with carbon. 18 I. Anisimov et al. The main gold losses during cyanidation were associated with invisible gold enclosed
in sulfides, scorodite and organic carbon. The main gold losses during cyanidation were associated with invisible gold enclosed
in sulfides, scorodite and organic carbon. Based on gold deportment results obtained, the following suggestions were made 1. Introduction of gravity in the grinding cycle would help reduce the loss of coarse gold
and extract some of the refractory gold with large particles of arsenopyrite and stibnite. Increase in recovery can vary as coarse gold content and be as much as 2–5%. 2. Addition of lead nitrate in cyanidation process may increase the dissolution of low-
grade gold particles. 2. Addition of lead nitrate in cyanidation process may increase the dissolution of low-
grade gold particles. 3. Flotation of carbon with consecutive tails leaching proved to have better recoveries
than direct leaching. Partial coal removing reduced preg-robbing on carbon for three
times and reduced gold losses by 22% from zone 1 ore. 4. The most effective objective to recovery would be liberation of minerals-carriers of
gold (mainly scorodite and arsenopyrite) from binaries with rock forming minerals. g
y
py
g
5. Leaching of gold from scorodite can be enhanced by NaOH pretreatment with
recovery increase of 10–15% for ore zone 1 and lesser effect for ore zone 2. 5. Leaching of gold from scorodite can be enhanced by NaOH pretreatment with
recovery increase of 10–15% for ore zone 1 and lesser effect for ore zone 2. 4
Conclusions Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Mineral-Geochemical Criteria to Gold
and Silver Recovery for Geometallurgical
Sampling Campaign on Primorskoe
Gold-Silver Deposit I. Anisimov(&), A. Sagitova, O. Troshina, and I. Agapov
Technology Research Department, Polymetal Engineering JSC,
St-Petersburg, Russia
anisimovis@polymetal.ru I. Anisimov(&), A. Sagitova, O. Troshina, and I. Agapov Technology Research Department, Polymetal Engineering JSC,
St-Petersburg, Russia
anisimovis@polymetal.ru Abstract. Ore variability study at Primorskoe gold-silver deposit demonstrated
wide variety of mineral composition and gold and silver recoveries with
cyanadation. The ore consisted of quartz-feldspar veins, quartz-rhodonite,
quartz-“pyrolusite”, quartz-epidote-garnet and quartz-Mn-silicates/hydroxides. Todorokite, birnessite, rancieite were the most common among the last ones. Statistical analyses of chemical and mineral composition, parameters of
cyanidation tests showed occurrence of three main ore types – feldspatic,
manganese silicate and oxide. Gold recovery effected by locking in Fe-oxides. The highest silver recovery strongly correlated with feldspathic cluster and
whiter sample color reflecting Ag mineral forms: acanthite and electrum. Ore
with silicate Mn showed good recoveries of acanthite, electrum and iodargirite
associated with Mn silicates. Main silver losses were connected with Mn-oxides
content and dark ore coloration where Ag chemically bound in Mn-oxides. Sr
and Ba content along with sample color were indications that could be used as a
proxy for recovery in geometallurgical mapping and ore-sorting. Keywords: Silver Predictive recovery Cyanidation
Manganese mineralization Keywords: Silver Predictive recovery Cyanidation
Manganese mineralization © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 19–23, 2019.
https://doi.org/10.1007/978-3-030-22974-0_5 2
Methods and Approaches 72 individual and 57 composite geometallurgical samples of Primorskoe deposit were
studied for chemical and mineral composition and cyanidation tests were performed. Ag varied from 176 to 36450 ppm in individual samples and gold – from 0.01 to
113 ppm. Ag content range in the composite samples was 49–1509 ppm averaging
606 ppm and Au – 0.04–9.92 ppm averaging 1.56 ppm. Chemical composition was studied at ALS labs, Moscow. Au content was assayed
by fire assay with atomic absorption finish, Ag and other 35 elements analyzed by ICP-
AES after four acid digestion. Quantitative X-ray powder diffraction by Rietveld refinement (QXRD) was carried
out using Bruker D8 Advanced diffractometer with Linxeye XE detector at Technology
Research Department, Polymetal Engineering. Statistical analysis was done on filtered data with Aitchison transformation using
PCA, regression analyses and Pearson correlations in Cytoscape software. 1
Introduction Primorskoe Au-Ag deposit is located in Omsukchan district of Magadan oblast, Russia. It formed by hydrothermal veins of various composition: Qu-Fsp-Chl with Ep, Qu-
Rdn-MnOx, Qu-MnOx and Qu-Gar ± Wol. Noble metal mineralization represented
with acantite, iodargirite, aurorite, jalpaite, pyrargyrite, electrum, kustelite and native
silver and gold. Abundance of specific manganese oxides effected gold and silver
extraction by cyanidation. The mineral composition study was aimed to define causes
of possible gold and silver losses with cyanidation tails and possible ways to recover
refractory silver. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 19–23, 2019. https://doi.org/10.1007/978-3-030-22974-0_5 © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 19–23, 2019. https://doi.org/10.1007/978-3-030-22974-0_5 I. Anisimov et al. 20 3
Results and Discussion Mineral composition of the sample significantly varied in quartz (up to 94%), feldspar
(up to 63%), micas (illite and muscovite – up to 25%). Epidote and piemontite formed
up to 14.5%, grossular and calderite – up to 8.9%. Bustamite (up to 22.8%), rhodonite
(up to 14.6%) and wollastonite (up to 16.7%) were observed in some samples. Apatite-
magnetite-titanite-vermiculite association was common in few samples. Manganese
oxides contents ran up to 34.4%. Mn-mineralization presented in 18 minerals: silicates (piemontite, calderite, bus-
tamite, rhodonite), carbonate (rhodochrosite), sulfide (alabandite), oxi-hydroxides. Todorokite, birnessite (7Å, 14-Å and amorphous), rancieite well spread among the last
ones. Manganite, pyrolusite, ramsdellite, jacobsite, bixbyite, pyrochroite, chalcophan-
ite, coronadite, aurorite were less common. Data population for multivariate statistical analyses included 103 following
parameters of chemical and mineral composition, color (RGB, brightness – BRT, and
darkness
-
DRN),
material
fineness
(c + 100),
silicate
and
oxide
Mn
ratio
(Mnsil/Mnox), 14 parameters of cyanidation conditions (NaCN concentration in the
pregnant solution – NaCNps; NaCNc and CaO consumption; final test pH – pHf; Au
and Ag contents in pregnant solution and cake – AuPS, AgPS, AuK, AgK; Au and Ag
recovery – eAu, eAg and losses – −eAu, −eAg (Fig. 1). Pearson correlations revealed
occurrence of two main geochemical and mineral clusters – whiter feldspathic (rock-
forming Ab-Olg-Chl) and darker manganese. Mgt-Tit-Ap-Vrm sub-cluster occurred in
the feldspathic one showing accessory syenite mineralization. “Skarn” association
(wollastonite, diopside, andradite, calcite, bustamite, rhodonite) and quartz vein (Qu,
SiO2, Cr and Iron – grinding contaminants) were distinguished. Ag recovery and NaCN
content in pregnant solution tied with color cluster. Mineral-Geochemical Criteria to Gold and Silver Recovery 21 COLOR
FELDSPATHIC
0,9
0,6
0,5
0,4
0,98
Legend
0,7
0,8
0,95
Positive
correlation with
Ag recovery
Positive
correlation with
Ag losses
MANGANESE
SKARN
QUARTZ
Mgt-Tit-
Ap-Vrm
Fig. 1. Correlations between 88 transformed parameters of individual and composite samples of
geometallurgical sampling campaign of Primorskoe deposit and proposed mineral association
clusters. Legend shows Pearson correlation coefficient values MANGANESE Fig. 1. Correlations between 88 transformed parameters of individual and composite samples of
geometallurgical sampling campaign of Primorskoe deposit and proposed mineral association
clusters. Legend shows Pearson correlation coefficient values Reagents consumptions (CaO, NaCN) correlated with Mn-cluster, proving that ion
exchange might take place during leaching. Zn, Cd, Pb tended to Mn-cluster without
connection to a specific Mn-oxide. 3
Results and Discussion Sr related to both main clusters equally reflecting
isomorphic distribution in feldspathic gangue as well as in MnOx. Reagents consumptions (CaO, NaCN) correlated with Mn-cluster, proving that ion
exchange might take place during leaching. Zn, Cd, Pb tended to Mn-cluster without
connection to a specific Mn-oxide. Sr related to both main clusters equally reflecting
isomorphic distribution in feldspathic gangue as well as in MnOx. Ag recovery demonstrated positive correlation with feldspathic cluster and sample
white color (BRT). Ag losses had strong connection with MnOx. Au recovery tied
together with color and quartz, Au losses correlated with MnOx and locking. PCA analysis showed same regularities. 6 principal components explained 54.78%
of the total variance and described mineral composition (Fig. 2), Mn- and accessory
mineralization, noble metal contents and recovery. PCA highlighted strong connection
between Ag loses, todorokite and amorphous birnessite. Regression Eq. (1) showed strong relation (R2 = 0.66) between silver recovery,
color (BRT) and some elements contents (in ppm). Variables listed in the order of their
significance: 22 I. Anisimov et al. Al2O3
Rb
Sc
Na2O
Ba
Zr
P
Olg
Fe2O3
Or
V
MgO
Sr
Ep
S
Co
-εAu
G
BRT
Act
R
B
εAg
-εAg
Ran
AgКек
NaCNк. Td
-BRT
εAu
AmBir
Prhr
Cld
Mn
AuПР
Au
Cr
SiO2
Qu
-4
-3
-2
-1
0
1
2
3
4
-4
-3
-2
-1
0
1
2
3
4
F2
F1
Bust-Wol
MnOx
Fsp
Qu
Qu-Fsp
Feldspathic-
muscovite with
MnOx
Mn-bearing
quartz veins
Quartz veins
(without MnOx)
Feldspathic-muscovite
without MnOx
Fig. 2. Factor loadings and factor scores for factors 1 and 2 with their interpretation Fig. 2. Factor loadings and factor scores for factors 1 and 2 with their interpretation eAg ¼ 18:24 þ 0:47BRT þ 1:39Al2O3 wt%
ð
Þ 0:12V þ 0:05Sr 3:45Sc
0:01Ag 1:21Mn wt%
ð
Þ þ 0:45Ca wt%
ð
Þ þ 0:003Cu 0:13Rb þ 0:01S
ð1Þ eAg ¼ 18:24 þ 0:47BRT þ 1:39Al2O3 wt%
ð
Þ 0:12V þ 0:05Sr 3:45Sc
0:01Ag 1:21Mn wt%
ð
Þ þ 0:45Ca wt%
ð
Þ þ 0:003Cu 0:13Rb þ 0:01S
ð1Þ eAg ¼ 18:24 þ 0:47BRT þ 1:39Al2O3 wt%
ð
Þ 0:12V þ 0:05Sr 3:45Sc
0:01Ag 1:21Mn wt%
ð
Þ þ 0:45Ca wt%
ð
Þ þ 0:003Cu 0:13Rb þ 0:01S
ð1Þ ð1Þ Mineral-Geochemical Criteria to Gold and Silver Recovery 4
Conclusions Individual and composite samples of Primorskoe Au-Ag deposit demonstrated variety
of mineral composition from high silica and feldspar to Mn-skarn association. Mn
mineralization presented in silicates, oxides and hydroxides, carbonate, sulfide had
great impact on Ag recovery. Wide range of vein gangue, ore and noble metal mineral
associations were similar to ones characteristic for Dukat deposit, the largest silver
deposit in Russia. Ag mineral forms affected recovery rate from highest presented by acanthite,
electrum in feldspatic association to the lowest locked in Mn-oxides, mainly in
todorokite and birnessite. Ore sorting by color and element content can be used. Mineral-Geochemical Criteria to Gold and Silver Recovery 23 23 Acknowledgements. The authors are grateful to Sergey Kubyshkin for performance of
cyanidation tests and Boris Milman for scientific advices. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. I. Anisimov(&), A. Sagitova, M. Kharitonova, A. Dolotova,
and I. Agapov Polymetal Engineering JSC, Saint-Petersburg, Russia
anisimovis@polymetal.ru Abstract. Levoberezhnoye gold deposit is located in Khabarovsk region. It
formed quartz-sulfide and quartz-adularia veining and fracture zones in argillic
altered intermediate volcanic tuffs and lavas. 61 variability study samples were
composed of quartz, feldspar, mica, kaolinite, chlorite with minor pyrite,
arsenopyrite, jarosite and accessories. Multivariate statistics of mineral com-
position and multi-element assays distinguished following ore types: (1) primary
quartz-feldspar sulfide-bearing breccia veins, (2) oxidized breccias with micas
transformed to illite-smectite; (3) high sulfidation quartz-kaolinite. Gold leach
recovery correlated with high sulfate content as well as mica and chlorite
transformation to illite and smectite. Low sulfidation ores showed lower
leaching recovery connected to gold encapsulation in pyrite. Thus, oxidized and
sulfate ore types were amenable to cyanidation, while primary ore was rec-
ommended for sulfide flotation gold recovery. Molybdenum high content con-
nected to Ag, Cu, Pb and As and supposed to be formed in a separate
mineralization event from gold. Keywords: Gold recovery Cyanidation Oxidation Illite-smectite Keywords: Gold recovery Cyanidation Oxidation Illite-smectite © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 24–28, 2019.
https://doi.org/10.1007/978-3-030-22974-0_6 2
Methods and Approaches 61 composite drill core sample of geotechnical mapping of Levoberezhnoe were
studied for cyanadation leaching and bulk mineral composition. Sample color was
described with RGB-parameters. Au was assayed with fire assay with atomic absorption finish, multi-element ICP-
AES assays after four acid digestion of straight and diluted samples and XRF-analysis,
sulfide and total S, total C estimated by LECO analysis. Mineral phase identification and their quantification was done using Eva software
and COD database. Quantitative X-ray powder diffraction with Rietveld refinement
Topas software at Polymetal Engineering. Multivariate statistical analysis was performed on filtered data with Aitchison
transformation using Pearson correlations with Cytoscape software, PCA and regres-
sion analyses. 1
Introduction Levoberezhnoye deposit is located in Khabarovsky region in Estern Russia. It is
localized in intermediate volcanics and formed steeply dipping quartz-adularia Au-Ag
breccia-vein system imbedded in rhyolites and extensively altered lake and flow tuffs
and ignimbrite volcanics. The ore bearing rocks suffered multiple hydrothermal brec-
ciation events with quartz±adularia-sulfide cement and fine sulfide dissemination in
altered volcanics. The veining and rocks are fine grained and hard for visual mineral
identification. The samples characterized with drastic variations in gold recovery by cyanidation
from 19 to 99%. The aim of the work was to determine compositional differences in ore
types and ore characteristics effected gold recovery and cased metal losses. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 24–28, 2019. https://doi.org/10.1007/978-3-030-22974-0_6 Mineralogical-Geochemical Criteria for Geometallurgical Mapping 25 3
Results and Discussion Three main ore types were distinguished based on mineral composition: 1 – quartz-
albite with sulfide (arsenopyrite-pyrite) mineralization, 2 – kaolinite-dickite with sul-
fate, 3 – illite-smectite. Wide structural and chemical variety of feldspars was observed: high and low
microcline, orthoclase, albite and hyalophane. Balancing mineral content QXRD
results for micas and K-felpdspars revealed significant potassium shortage and sug-
gested high baddingtonite content in feldspar, which needs confirmation with assays. Observed anomalously high values of molybdenum were connected to Ag, Pb, Au and
Cu. Multivariate statistics analyses included 73 following parameters: chemical and
mineral composition, color (RGB, brightness – BRT and darkness - DRN), material
fineness (c + 100), Au recovery – eAu and losses – −eAu. Pearson positive correla-
tions revealed occurrence of 5 geochemical and mineral clusters (Fig. 1): 1 – quartz-kaolinite-albite-sulfate with zeolites, Sr enriched (high sulfidation
alteration); 2 – micas-chlorite with calcite group was linked to 3 - sulfide mineralization
(beresite – low sulfidation); 4 – potassic feldspar-ankerite; 5 - oxidation cluster between
kaolinite and sulfide/micas. 1 – quartz-kaolinite-albite-sulfate with zeolites, Sr enriched (high sulfidation
alteration); 2 – micas-chlorite with calcite group was linked to 3 - sulfide mineralization
(beresite – low sulfidation); 4 – potassic feldspar-ankerite; 5 - oxidation cluster between
kaolinite and sulfide/micas. Qu-Kln-Ab cluster showed kaolinitization. K-Fsp and mica clusters tied together
with K and Rb. Barium feldspar - hyalophane associated with K-Fsp. Mica cluster
combined muscovite, illite, illite-smectite and chlorite. Sulfide cluster connected with
mica cluster and gold losses. Au recovery connected with As, illite, illite-smectite
indirectly through the minerals formed from sulfide oxidation: jarosite, goethite and
scorodite. Mica transformation to illite and illite-smectite followed oxidation of sulfides 0,5
0,8
0,7
0,4
0,3
0,9
Positive correlation
with Au recovery
Positive correlation
with Au losses
Legend
SULFIDE
K-FSP
εAU
WHITE MICAS
QTZ-
KLN-AB
Fig. 1. Pearson correlations of 74 transformed chemical, mineral composition and cyanidation
tests parameters of 51 small composite samples from Levoberezhnoye deposit
26
I. Anisimov et al. I. Anisimov et al. 26 Fig. 1. Pearson correlations of 74 transformed chemical, mineral composition and cyanidation
tests parameters of 51 small composite samples from Levoberezhnoye deposit and, thus reflecting good Au leach results. Beresite group coupled with gold losses, the
second ones indirectly correlated with gold recovery. PCA analysis exposed 6 principal components, which explained 58.14% of the total
variance. They described mineral composition (Fig. Mineralogical-Geochemical Criteria for Geometallurgical Mapping 27 27 Be
Fe
Mg
Ms
Ill
DRN
K
IS
Co
Zn
Mn
Rb
Ca
S
Jrs
Scr
Py
-εAu
Sslfd/Stotal
C
Ox/S2-
S2-/Ox
εAu
Gth
R/BRT
Zeo
P
Ba
Kln
Pl
BRT
R
Dsp
G
Hcl
SrP
Sr
-4
-3
-2
-1
0
1
2
3
4
-4
-2
0
2
4
F3
F1
K-Feldspar
Kaolinite
Illite-Smectite
Scorodite-
jarosite
Sulfide
Kaolinite-
plagioclase
Fig. 2. Factor loadings and factor scores for factors 1 and 3 with interpretation 4
F1 Fig. 2. Factor loadings and factor scores for factors 1 and 3 with interpretation 3
Results and Discussion 2), oxidation rate, rare-metals and
arsenic associations, color, grinding fineness. Results were similar to ones obtained
with pair correlations: Au losses linked to sulfides, oxidation rate raised Au recovery
(Fig. 2). Thus, flotation would be the best Au recovery solution from primary sulfide
ore and tank cyanidation to oxide one. Regression equation for gold recovery by cyanidation was calculated. It had rela-
tively low R2 = 0.46 (1) and connected color parameter (R/BRT) and elements contents
(ppm and wt%): eAuCN ¼ 96:91 þ 185:3R
BRT 7:83Ti %
ð
Þ 2:71Co 0:73La 1:15Pb þ 0:53Sc 0:004Zr þ
1:81Ga 13:09Ssfd %
ð
Þ þ 10:90Fe %
ð
Þ 0:54V þ 0:34Ag 1:94Ni 75:18Ca %
ð
Þ þ
0:01Ba þ 0:01P eAuCN ¼ 96:91 þ 185:3R
BRT 7:83Ti %
ð
Þ 2:71Co 0:73La 1:15Pb þ 0:53Sc 0:004Zr þ
1:81Ga 13:09Ssfd %
ð
Þ þ 10:90Fe %
ð
Þ 0:54V þ 0:34Ag 1:94Ni 75:18Ca %
ð
Þ þ
0:01Ba þ 0:01P ð1Þ ð1Þ I. Anisimov(&), A. Dolotova, A. Sagitova, M. Kharitonova,
and I. Agapov Polymetal Engineering JSC, Saint-Petersburg, Russia
anisimovis@polymetal.ru Abstract. Varvara Au-Cu mine deposit is located in Northern Kazakhstan. Mineralization is hosted in volcano-sedimentary, sedimentary rocks, metamor-
phosed and altered ultramafic and felsic rocks. Variability study was done on 58
composite samples represented five mineral ore types: serpentine-chlorite-talc;
carbonate-chlorite-talc;
quartz-sulfide;
pyroxene-chlorite-prehnite ± garnet;
quartz-feldspar ± pyroxene ± amphibole. Five processing ore types were
defined: Au, Ni-As, pyrite, Cu and mixed. Mineralogy and geochemical studies
revealed separate mineral associations carrying Cu, Ni-As and Au mineraliza-
tion. Flotation and cyanidation tests were performed for each sample. Au losses
with cyanidation cake occurred due to locking in sulfides. Floatation concentrate
contamination with Mg-silicates (talc, serpentine) was connected to Au losses. Quartz-sulfide ore demonstrated better recovery by flotation. Cyanidation were
most effective for pyroxene-chlorite-prehnite ± garnet and quartz-feldspar ±
pyroxene ore compositions. Carbonate presence in the serpentine-chlorite-talc
ore followed decrease in recovery by both extraction methods. Optimal viable
ore treatment method can be chosen based on regression equations using ore
chemical composition and color. Keywords: Typification Cyanidation Flotation Recovery estimation
Geometallurgy Keywords: Typification Cyanidation Flotation Recovery estimation
Geometallurgy Mineralogical Reasons of Au Recovery
Variability from North-Western Pit of Varvara
Au-Cu Mine (Kazakhstan) and Criteria
for Geometallurgical Mapping I. Anisimov(&), A. Dolotova, A. Sagitova, M. Kharitonova,
and I. Agapov 4
Conclusions Mineral composition of the samples varied significantly from high quartz (up to 70%)
and feldspar (up to 64%) to kaolinite (up to 45%) and illite-smectite (up to 40%). Au
losses with cyanidation tails were bound to sulfides content. Sulfide oxidation to jar-
osite and scorodite and red component of sample color reflected increase in Au
recovery. Absence of chlorite and transformation of muscovite to illite-illite-smectite
also manifested Au cyanadation recovery improvement. Revealed links between samples characteristics and regression equation, geomet-
allurgical mapping and ore sorting can be performed based on sample color and
chemical composition (Ti, Co, La, Pb, Sc, Zr, S, Fe, V, Ag, Ni, Ba). 28 I. Anisimov et al. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches 58 small composite geometallurgical samples from Northwestern open pit were studied
for bulk chemical and mineral composition, sample color with RGB-parameters. Cyanidation and sulfide flotation tests were done on the head samples. Au was assayed with fire assay with atomic absorption finish, multi-element ICP-
AES assays after four acid digestion of samples and XRF-analysis, sulfide and total S,
total C estimated by LECO AES analysis. Mineral phase identification and their quantification was done using Eva software
and COD database. Quantitative X-ray powder diffraction with Rietveld refinement
was done using Topas software at Polymetal Engineering. Multivariate statistical analysis was performed on filtered data with Aitchison
transformation using Pearson correlations with Cytoscape software, PCA and regres-
sion analyses. Regression analysis was carried out to predict gold extraction. 1
Introduction Varvarinskoe Au-Cu deposit is located in Kostanay region in Northern Kazakhstan and
operated by several open pits. It is localized in volcanogenic, terrigenic, carbonate
rocks (D2–C1) with ultrabasic and granodiorite intrusions (C2–C3). Silicification,
argillic alteration, scarn processes were wide spread. Retrograde metamorphic changes
were typical for ultramafic rocks. This work was aimed at determining compositional differences in ore types and ore
characteristics effected gold recovery by cyanidation and flotation. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 29–32, 2019. https://doi.org/10.1007/978-3-030-22974-0_7 30 I. Anisimov et al. 3
Results and Discussion Mineral composition of the samples varied between serpentine-talc-chlorite ±
carbonate, quartz-feldspatic and quartz-sulphide associations. Five main mineral ore
types were distinguished based on bulk mineral composition: 1 – serpentine-chlorite-
talc, 2 – carbonate-chlorite-talc, 3 – quartz-sulfide, 4 – pyroxene-chlorite-prehnite-
garnet, 5 – quartz-feldspar-pyroxene-amphibole. Flotation and leaching tests revealed five processing ore types: a - gold, b - copper,
c - Ni-As, d - pyrite and e - mixed. c - Ni-As, d - pyrite and e - mixed. Multivariate statistics analyses were performed on all available samples parameters
including mineral and chemical composition; color (RGB, brightness – lBRT and
darkness - dBRT); material fineness (−71 µA); Au, Cu, As, Ni, sulfidic S, total S
recovery to concentrate and losses to flotation tails (XF) (eAuKF, eCuKF, eAsKF,
eNiKF, eSsKF, eSKF; eAuXF, eCuXF, eAsXF, eNiXF, eSsXF, eSXF) and cyanidation
(eAuCN, eCuCN, −eAuCN, −eCuCN), parameters of cyanidation conditions (reagents
uptake - NaCNi, CaCNi), cyano-soluble copper – aCuCNr, Au content in cake –
AuXvA, characteristics of gravity con (mass pull - cgk-t, pyrite, arsenopyrite, Cr-
spinel, chalcopyrite, Ni-minerals contents in gravity con: PyMrcGK, ApyGK, ChrGK,
CpyGK, NiMGK). Pearson correlations revealed occurrence of 3 geochemical and mineral clusters
following bulk sample mineralogy (Fig. 1): quartz-sulfide, diorite (feldspatic) and
serpentinite. Au losses with cyanidation tails connected with Au locked in sulfides,
which was proved with SEM study. S and Cu contents and NaCN consumption were
included in the cluster with Au losses with cyanidation. This fact may point to con-
sumption of free CN−by reaction with S0 and Cu dissolution, thus leading to deficit of
free CN−. This reaction produced CuCN and rhodanates that could block surface gold
particle impeding its dissolution. Concentrate contamination with self-floating Mg-
silicates (talc, serpentine) was connected to Au losses with flotation tails. Principal component analysis exposed 7 principal components, which explained
72.1% of the total variance and described mineral composition, geochemical Mineralogical Reasons of Au Recovery Variability 31 Quartz-sulfide
Diorite (feldspatic)
Serpentinite
Correlation coefficients
Positive correlation with Au recovery by Cyanidation
Positive correlation with Au losses with Cyanidation tails
Fig. 1. Positive correlations between compositional and metallurgical parameters of samples Diorite (feldspatic) Positive correlation with Au recovery by Cyanidation
Positive correlation with Au losses with Cyanidation tails Positive correlation with Au recovery by Cyanidation Positive correlation with Au recovery by Cyanidation
Positive correlation with Au losses with Cyanidation tails Positive correlation with Au losses with Cyanidation tails Fig. 1. 3
Results and Discussion Positive correlations between compositional and metallurgical parameters of samples
from Varvara Fig. 1. Positive correlations between compositional and metallurgical parameters of samples
from Varvara Fig. 1. Positive correlations between compositional and metallurgical parameters of samples
from Varvara associations, Au and Cu recovery by flotation and cyanidation (Fig. 2). 5 ore types
were well separated in the coordinates of factors 2 and 3, 2 and 4. Quartz-sulfide ore
samples demonstrated better recovery by flotation, while cyanidation was more
effective for samples with pyroxene-chlorite-prehnite-garnet and quartz-feldspar-
pyroxene-amphibole compositions. Carbonate occurrence in the serpentine-chlorite-
talc ore reduced recovery by both concentrating methods. Regression equations for Au recovery by flotation (R2 = 0.80) and by cyanidation
(R2 = 0.88) included color parameter (BRT) and elements contents (ppm or wt%): eAuFl ¼ 127:32 þ 0:17Co 0:01Cu 4:78Fe %
ð
Þ þ 5:19K %
ð
Þ þ 0:39Mo þ 0:39Th þ 0:01Sb
0:63BRT þ 0:10Zn þ 0:43Mg %
ð
Þ þ 0:84SiO2 %
ð
Þ 0:81Li 0:40Rb 1:97Te þ 5:03Cs eAuCN ¼ 226:60 2:86Ca %
ð
Þ þ 0:30Co 7:00Fe %
ð
Þ 2:08Sc 0:16Sr þ 0:13V
0:24MgO %
ð
Þ 3:37Al2O3 %
ð
Þ 0:55SiO2 %
ð
Þ þ 1:98Ga 0:42BRT þ 0:27LOI %
ð
Þ þ
0:004Cu þ 0:02Ni 0:01As 32 I. Anisimov et al. Ce/Ce*
SiO2
Py
BaO
Tl
S
Rb/Sr
CaCNi
εAuCN
Prx
R
εAuXF
lBRT
GB
εNiKF
Qu
Chl
εNiXF
Sb
Ca
γgk-t
εAuKF
dBRT
Gy
Fe
Cu
Eu/Eu*
-εAuCN
MnO
Calc
NaCNi
MgO
-4
-3
-2
-1
0
1
2
3
4
-4
-3
-2
-1
0
1
2
3
4
F4
F2
Srp-Chl-Tlc
Carb-Chl-Tlc
Qu-Sulf
Prx-Chl-Pre with Grt
Qu-Fsp
εAu by flotation and Au
losses with cake
Au losses with flotation tails
and cake
εAu by cyanidation and
Au losses with flotation tails
εAu by flotation and
cyanidation
Fig. 2. Factor loadings and factor scores for factors 2 and 4 with interpretation Fig. 2. Factor loadings and factor scores for factors 2 and 4 with interpretation © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 33–36, 2019.
https://doi.org/10.1007/978-3-030-22974-0_8 4
Conclusions Mineral composition of the samples varied significantly from serpentine-talc-chlorite ±
carbonate to quartz-feldspar, skarn and quartz-sulfide associations. Au losses with
cyanidation tails connected with Au locked in sulfides. Au losses with flotation tails
connected to concentrate dilution with self-floating Mg-silicates (talc and serpentine). Ore sorting can be done based on regression equations of element composition and ore
color to direct the ore to more economically viable process. Acknowledgements. The authors are grateful to Nicolay Rylov for scientific advices, Lims-lab
and Nati-R for collaboration. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Mineralogical Breakthrough into Nanoworld:
Results and Challenges A. Askhabov(&) Institute of Geology Komi SC UB RAS, Syktyvkar, Russia
askhabov@geo.komisc.ru Abstract. A bit more than a hundred years ago Wolfgang Ostwald, Professor at
the University of Leipzig, published a book titled “The World of Neglected
Dimensions” (Ostwald 1923). He announced a program of a research break-
through into the world of microscopic particles, which was imminent by that
time. A new stage of the intervention into “world of neglected dimensions”
began near the end of the 20th century. The main objects at this stage were
nanosized particles. The agenda raised issue of development of new sciences,
including nanomineralogy. The subsequent mineralogical intervention into the
“world of neglected dimensions” proved to be quite successful. We associate
challenges of the next breakthrough with the study of objects and processes in
the range from individual atoms and molecules to the first mineral individuals
(nanoindividuals). The protomineral world is today a new “world of neglected
dimensions”. Keywords: “world of neglected dimensions” Nanoparticles
Quatarone concept Protomineral world Keywords: “world of neglected dimensions” Nanoparticles
Quatarone concept Protomineral world 1
Introduction The unprecedented interest in nanoscale objects, which we witness in recent years, I
called a new stage of intervention into the “world of neglected dimensions”. The first
stage of the intervention (research breakthrough) occurred at the beginning of the 20th
century and was associated with W. Ostwald, who actually referred to the region of
microscopic particles as “the world of neglected dimensions” (Ostwald 1923). Inten-
sive researches in this area, which began then, resulted in formation of a new science -
colloid chemistry. Its creators R. Zigmondi (“he opened access to the world of inac-
cessible sizes”), T. Svedberg and J. Perrin (for “a breakthrough into the world of
discrete particles”) were awarded the Nobel Prizes in 1925 and 1926. What is hap-
pening in our time is a secondary discovery of the “world of neglected dimensions”,
now not only at micro, but also at nanoscale. Mineralogical intervention into the micro and nanoworld began long ago. There is
generally nothing revolutionary in what is happening nowadays. Nano-mineralogy is a
normal and inevitable stage in the development of mineralogical science. Moreover the
role of mineralogy (crystallography) in the study of nanoscale objects is quite com-
parable with the role of physics, chemistry or biology. Suffice it to recall that structural
mineralogy always operated on nanoscale elements, and nucleation and growth of A. Askhabov 34 crystals is a typical nanoprocess, a crystalline nucleus is a nanocrystal, and opal is a
nanostructured natural material. Nanotechnology is often a repetition of natural pro-
cesses or nature-like technologies. The very first results of the mineralogical intervention in the nanoworld were very
impressive (Nanomineralogy… 2005). Among the most important achievements of the
past years: – - discovery of a new type of structurally and morphologically ordered objects –
nanoindividuals. The likely morphological diversity of nanoindividuals is enormous
and not limited by laws of classical crystallography; – - significant expansion of boundaries of mineral world due to solid amorphous
substances previously attributed to mineraloids. A discovery of a new class of
structurally ordered mineral structures; – - finding of the lower limit of mineral objects, beyond which matter is in a different,
non-mineral (protomineral, quatarone, cluster) state; – - identification of common laws of self-organization at nanoscale in the mineral and
living worlds. Mineral and living matter, as is known, are formed at nanoscale. 1
Introduction Both
do not exist outside the lower limit of the nanolevel; – - substantiation of fundamental role of forms of existence and properties of
nanoscale particles in minerals and ores for the development of new technologies
for deep and complex processing of mineral raw. The research of the mineral nanoworld is just beginning. The study of mineral
nanostructures, natural clusters, nanostructured natural materials, organo-mineral
nanoobjects, nanodispersed phases in ores, development of new technologies for
their extraction, modifying properties of minerals and mineral nanoparticles is an
incomplete list of the nearest tasks of general and applied nanomineralogy. At that we
will have to reconsider a number of fundamental concepts, significantly expand them or
introduce new ones. Another argument for the expansion of mineralogical objects to
nanoscale is that on this basis a great challenge for mineralogical science can be
formed, which is now missing. It is not reasonable to reject a breakthrough promising
unprecedented discoveries and a deeper understanding of nature of the mineral matter
and its sources. It is hard to deny the intellectual appeal and charm of the nanomin-
eralogical project. The opening door to the nanoworld should not be closed, even if
there is a threat of erosion of foundations of classical mineralogy. Significant progress in understanding properties of the nanoworld was made by the
quatarone concept of cluster self-organization of matter developed by us (Askhabov
2011). Within this concept new models of crystal nucleation (Askhabov 2016), for-
mation of various types of nanoparticles (including fullerenes (Askhabov 2005a)) and
solid amorphous materials (Askhabov 2005b) are proposed. At that quatarons them-
selves are new nanoobjects without analogues in the macroworld. They are not small
pieces, cut from a large piece of matter or obtained by successive division. They cannot
be identified with ordinary clusters—equilibrium structures optimized by geometry or
energy. The quataron concept solved the fundamental problem of deciphering the mecha-
nism of crystal growth in an amazingly simple way (Askhabov 2016). Quatarons
proved to be ideal building units for crystal growth. Due to the dynamism of structure, Mineralogical Breakthrough into Nanoworld: Results and Challenges 35 their inclusion into the crystal lattice occurs with virtually no kinetic resistance and
deformation of the crystal lattice. The quataron model of crystal growth acts as an
alternative to the known models of crystal growth by attaching individual atoms or
ready crystalline blocks. 1
Introduction The development of ideas of the quataron concept opened a window to the pro-
tomineral world. And this world is today regarded as a new “world of neglected
dimensions” (Askhabov 2017). As a result, the program of W. Ostwald receives a new
impetus and focuses on the study of objects of the protomineral world. We must answer
not only the questions of how minerals are formed, but also why they are formed, why
minerals are exactly as they are. The ontogenesis of minerals should begin not with the
origin of the mineral, but with the protomineral state of mineral matter. Thus, the world of minerals is preceded by the world of specific pre-mineral objects
—the protomineral world (Fig. 1). This world requires interdisciplinary approaches and
new instrumental methods with spatial angstrom-nanometer and temporal femto-
nanosecond resolution. In connection with the emergence of the protomineral world in
the current agenda of mineralogical science, it is highly desirable to draw up a program
of top-priority experiments for implementation with the European free electron laser - a
device that is potentially able to satisfy requests of mineralogical science and study in
detail the process at the source of mineral matter. Fig. 1. Way from the world of separate atoms and molecules toward the world of mineral
individuals comes through the protomineral world. Fig. 1. Way from the world of separate atoms and molecules toward the world of mineral
individuals comes through the protomineral world. Acknowledgements. The work was accomplished with partial financial support by UB RAS
program No. 18-5-5-45, and RFBR No. 19-05-00460a. Acknowledgements. The work was accomplished with partial financial support by UB RAS
program No. 18-5-5-45, and RFBR No. 19-05-00460a. A. Askhabov 36 1
Introduction Since Chile is the largest Cu producing country with the biggest reserves, Chilean
mining industry generates huge quantities of mining residues, amongst others in the
form of tailings dumps. The fact that some of them were generated many decades ago,
where process technology was inadequate in comparison to today, makes some of them
economically interesting. Due to advanced weathering, the tailings, which are poten-
tially acid producing, bear an environmental hazard. Therefore, our aim is to investigate
the geochemistry and mineralogy of the tailings in order to determine their economic
potential and environmental impact. The studied mine tailings dump is located in the region of Coquimbo, where an arid
Mediterranean climate with mean annual temperature of 14.6 °C, precipitation of
132 mm, and evaporation of about 1702 mm prevails (Mora et al. 2007). Geologically,
the region is characterized by Early Cretaceous Chilean Manto-type (volcanic-hosted
stratiform) Cu deposits (Kojima et al. 2003) that are the most probable source of the
studied tailings. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 37–40, 2019.
https://doi.org/10.1007/978-3-030-22974-0_9 Case Study: Geochemistry and Mineralogy
of Copper Mine Tailings in Northern
Central-Chile K. Berkh(&), D. Rammlmair, M. Drobe, and J. Meima Abstract. Selected mine tailings in northern-central Chile were geochemically
and mineralogically studied for their economic potential and environmental
impact. High bulk Co content up to 1500 ppm and Cu content up to 9100 ppm
are caused by Co-bearing pyrite, chalcopyrite, and their secondary products such
as malachite and Co-Cu-carrying Fe-hydroxides. Due to high amount of sulfide
minerals acid mine drainage (AMD) is forming in the oxidized upper part of the
tailing, which makes a retreatment in dispensable to reduce the environmental
impact. Keywords: Mine tailings Geochemistry Mineralogy Cobalt Copper
Reprocessing Keywords: Mine tailings Geochemistry Mineralogy Cobalt Copper
Reprocessing Case Study: Geochemistry and Mineralogy
of Copper Mine Tailings in Northern
Central-Chile
K. Berkh(&), D. Rammlmair, M. Drobe, and J. Meima
Federal Institute for Geosciences and Natural Resources, Hanover, Germany
Khulan.berkh@bgr.de Case Study: Geochemistry and Mineralogy
of Copper Mine Tailings in Northern
Central-Chile
K. Berkh(&), D. Rammlmair, M. Drobe, and J. Meima
Federal Institute for Geosciences and Natural Resources, Hanover, Germany
Khulan.berkh@bgr.de References Ostwald W (1923) World of neglected dimensions. Introduction to modern colloid chemistry
with an overview of its applications. Mir, Moscow, 228 p (in Russian) Yushkin NP (2005) Ultra- and microdispersed state of the mineral matter. In: Askhabov AM
Rakin VI (eds) Nanomineralogy. Nauka, St. Petersburg, 581 p (in Russian) Yushkin NP (2005) Ultra and microdispersed state of the mineral matter. In: Askhabov AM,
Rakin VI (eds) Nanomineralogy. Nauka, St. Petersburg, 581 p (in Russian) Askhabov AM (2011) Quataron concept: main ideas, some applications. In: Proceedings of komi
science center UB RAS.- №3, no 7, pp 70–77 (in Russian) Askhabov AM (2016) Quataron crystal nucleation and growth models. In: Proceedings of RMS.-
Ch. CXLV, no 5, pp 17–24 (in Russian) Askhabov AM (2005a) The kvataron model of fullerene formation. In: Physics of solid state, vol
47, no 6, pp 1186–1190 Askhabov AM (2005b) Aggregation of quatarons as a formation mechanism of amorphous
spherical particles. In: Doklady earth sciences, vol 400, no 1, pp 937–940 Askhabov AM (2017) New stage of mineralogical invasion into the “world of neglected
dimensions”: discovery of the protomineral world. In: Proceedings of anniversary congress of
the Russian mineralogical society “200th anniversary of the Russian mineralogical society”,
St. Petersburg, vol 2, pp 3–5 (in Russian) Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches Eight drill cores with lengths of seven meters were taken from the tailings dump
(Fig. 1). Bulk geochemistry of homogenized material for each meter was investigated
using
a
standard
WDXRF. Mineralogy
was
studied
on
representative
grain K. Berkh et al. 38 concentrates obtained by gravity separation. Abundance of the minerals was analyzed
by MLA and composition of the minerals was examined by EPMA. Fig. 1. Drill cores positions (M1–M8) in three heaps (I-III) Fig. 1. Drill cores positions (M1–M8) in three heaps (I-III) 3
Results and Discussion The tailings material consists of alternating layers of sand, silt and clay. According to
geochemical pattern, the tailings dump can be subdivided into three groups, as shown
in Fig. 1. Heap I and II are enriched in Fe and Co (Fig. 2). Special feature of heap I is
a depletion in S and Ca but a high percentage of loss on ignition (LOI). It may point to
a weathered part of the tailings dump, where oxidation of pyrite results in dissolution of
calcite and accumulation of water bearing secondary clay minerals. Heap III contains
high amounts of host rock and therefore elevated contents of Si, Al, Mg, Na, K and P
can be seen. It is strongly enriched in Cu but only at the near surface level (Fig. 2). Fig. 2. Bulk content of relevant elements in three heaps Fig. 2. Bulk content of relevant elements in three heaps Primary gangue minerals in heap I and II are epidote, quartz, andradite, and albite
pointing towards skarn mineralization. The only difference between the two heaps is the
absence of calcite in the heap I confirming the bulk geochemistry. The most common
primary ore mineral in both heaps is pyrite, which occurs as liberated grains. Three types
of pyrite were identified. The first type is pure pyrite. The second type has an As-rich rim
with up to 8.7 wt% As (Fig. 3a) and is predominantly present in the heap II resulting in Case Study: Geochemistry and Mineralogy of Copper Mine Tailings 39 an elevated bulk content of As (Fig. 1). The third type contains a significant amount of
Co. Hereby, the Co concentration increases from rim to core and can reach up to 3.8 wt
%. Additional trace elements are e.g. up to 0.5 wt% Ni, 0.1 wt% Cu, and 0.2 wt% Cd. Remaining primary ore minerals are magnetite, hematite and trace amounts of chal-
copyrite. They also occur as well liberated grains and are occasionally inter grown with
each other. These minerals do not contain significant amounts of trace elements. Fig. 3. BSE images of the pyrite grains: a As-rich rim on pyrite and b pyrite rimmed by Fe-
hydroxide with significant amounts of Co and Cu Fig. 3. 3
Results and Discussion BSE images of the pyrite grains: a As-rich rim on pyrite and b pyrite rimmed by Fe-
hydroxide with significant amounts of Co and Cu Secondary phases are gypsum and Fe-hydroxides as weathering products of calcite
and pyrite (Fig. 3b) and preferentially occur in heap I. Such in-situ precipitation of Fe-
hydroxides can only take place at near neutral pH proving an initial occurrence of
calcite as a buffering agent in the strongly weathered heap I. The Fe-hydroxides contain
high concentrations of Co (up to 0.9 wt%), Cu (up to 2.5 wt%) and Ca (up to 4.3 wt%),
pointing to a dissolution of pyrite, chalcopyrite and calcite and fixation of released
metals in Fe-hydroxides. The analyzed saturated soil extraction had a pH of 2. Its Co
and Cu contents were 103 and 136 mg/l respectively, whereas Hg, Cd, As, Zn and Pb
contents remained under 2.4 mg/l. Especially, the Co and Cu values exceed the
international guidelines (IFC 2007 and Gusek and Figueroa 2009) by more than a
factor of 100.In fact, the potential for AMD is high in the oxidized parts of the heaps. The heap III predominantly consists of gangue minerals. The most common
mineral is quartz followed by albite, anorthite, micas, K-feldspar, amphibole, epidote,
pyroxene, chloride, and calcite. A Minor amount of Fe-oxides and chalcopyrite exists,
either as liberated grains or finely intergrown with gangue minerals. Chalcopyrite is
strongly replaced by Fe-hydroxides and Cu-carbonates along grain boundaries and
micro fractures. 4
Conclusion Bulk geochemistry provides a good prediction of the mineralogy. Elevated amounts of
Fe, Co, and S in the heaps I and II correspond to Co-bearing pyrite-rich waste material. In contrast, heap III consists of host rock forming elements resulted by non-sulfidic
gangue mineral waste. From an economic point of view, Co is the only valuable metal K. Berkh et al. 40 in the heap I and II. For the extraction magnetic separation should be performed to
eliminate high amounts of magnetite. Heap I contains extensive amounts of Co- and
Cu-rich Fe-hydroxides that usually coat pyrite grains. Therefore, leaching can be
directly applied to extract acid soluble Co and Cu and also to liberate pyrite grains. Otherwise, pyrite cannot be floated. Afterwards, pyrite from both sulfidic heaps can be
floated and Co can be extracted by bioleaching. In case of heap III, only the uppermost
first meter, which is Cu-rich, should be treated. Cu occurs preferentially as acid soluble
mineral such as malachite and Cu-rich Fe-hydroxides. However, calcite would have to
be removed by gravity separation to reduce the consumption of sulfuric acid. Other-
wise, itis economically not feasible. From an environmental point of view, heap III
poses no environmental risk because it does not host AMD potential. Oxidation of
minor chalcopyrite will be buffered by the carbonate content. In turn, the sulfidic
tailings should be immediately treated, because the potential of AMD generation due to
heavy rains is high. The hazardous potential of the impoundment should not be
underestimated because of agricultural activities in the vicinity. Acknowledgements. This investigation is supported by DERA the German Mineral Resources
Agency of BGR. Acknowledgements. This investigation is supported by DERA the German Mineral Resources
Agency of BGR. References Gusek JJ, Figueroa LA (2009) Mitigation of metal mining influenced water (Management
Technologies for Metal Mining Influenced Water, vol 2), Soc Min Metall Explor. Littleton,
Colorado IFC (2007) Environmental, health and safety guidelines for mining. International Finance
Corporation, World Bank Group, DC, 33 p Kojima S, Astudillo J, Rojo J, Tristá D, Hayashi K (2003) Ore mineralogy, fluid inclusion, and
stable isotopic characteristics of stratiform copper deposits in the coastal Cordillera of
Northern Chile. Miner Deposita 38:208–216 p
Mora F, Tapia F, Scapim SA, Martins EN (2007) Vegetative growth and early production of six
olive cultivars, in Southern Atacama Desert, Chile. J Cent Eur Agric 8(3):269–276 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Application of Fluoride Technology
for Processing of Off-Grade Aluminum
Raw Materials I. Burtsev, I. Perovskiy(&), and D. Kuzmin Institute of Geology named after Academician N.P. Yushkin Komi Science
Center of the Ural Branch of the Russian Academy of Sciences,
Syktyvkar, Russia
igor-perovskij@yandex.ru Abstract. We used the process of hydrofluoride desilication for off-grade
aluminum raw materials widely distributed in the Komi Republic. The optimal
ratios of fluorinating agent (NH4HF2) to the target component (SiO2) were
determined by the method of differential thermal analysis. At a fluorination
temperature 300 °C and timing 30 min, we obtained concentrates containing up
to 80% Al2O3 and up to 10% SiO2. Keywords: Bauxite Aluminum raw materials Desilication Fluorination
Ammonium hydrofluoride © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 41–44, 2019.
https://doi.org/10.1007/978-3-030-22974-0_10 2
Methods and Approaches The objects of research - representative samples of bauxite, allite, kaolinite clays from
the Vezhayu-Vorykvinskoe (V-V), Verkhneshchugorskoe (Vsch) deposits and occur-
rences of the Izhma area (IA). Samples were grinded in a disk eraser (ID-200) and
separated by size classes 0.125–0.250 and 0.25–0.5 mm. Analytical work was carried
out by the equipment of the Center for Collective Use GeoScience of the Institute of
Geology Komi SC UB RAS using XRD, XRF, DTA. Fluorination was carried out in a
tube furnace equipped with a gas-extraction system. Ammonium hydrodifluoride
(NH4HF2) was used as a fluorinating component, which is an ecologically safe matter
under standard conditions. 1
Introduction One of the most important problems of Russian aluminum industry is the deficit of
high-quality alumina raw materials, forcing to import it from other countries. A major source of alumina raw materials is the Vezhayu-Vorykvinskoe and
Verkhneshchugorskoe bauxite deposits, which are part of the Vorykvinskaya group of
deposits in Middle Timan and developed by the United Company RUSAL. Mined
bauxites are predominantly consumed for alumina production by the Bayer method. The share of low-quality sintered bauxites is significant and amounts from 5 to 55%
of balance reserves for Vezhayu-Vorykvinskoe and Verkhneshchugorskoe deposits. A sharp reduction in consumption of sintered bauxites makes an alternative processing
a rather actual question. In addition, the prospected reserves contain a significant
amount of off-balance ores, off-grade in quality: high-silica low-modulus (better MSi
2.6–3.8) bauxites and associated high-aluminous allites. Gravitational, flotation, magnetic, chemical, and other methods have been proposed
to enrich the bauxites, but their use for enriching Middle Timan bauxite is limited due
to a fine size of the minerals, a large proportion of amorphous phases, and a small
efficiency of used processes in general (Burtsev et al. 2016). Fluoride technologies are one of the most promising ways to process mineral raw
materials (Dyachenko and Kraidenko 2007; Medkov et al. 2011). The purpose of the
research – to assess the prospects for the use of fluorination to desilicate off-grade I. Burtsev et al. 42 aluminum raw materials (bauxite, allite, kaolinite clays), which are widespread in the
Komi Republic. SiO2 þ 3NH4HF2 ¼
NH4
ð
Þ2SiF6 þ 2H2O þ NH3 " We studied the process of fluorination of off-grade bauxites at molar ratios SiO2:
NH4HF2 equal to 1: 1 and 1: 1.5 with the help of DTA. We established that ratio 1:1
was preferred. Excessive ammonium fluoride (1:1.5) did not result in a positive effect,
but was accompanied by passivation of fluorination reactions and formation of a larger
volume of gaseous products. Taking into account DTA data, the temperature regime of fluorination consisted of
sintering the sample with NH4HF2 at temperature 200 °C (30 min.) with subsequent
sublimation of resulting salt (NH4)2SiF6 at 300 °C (30 min.). Upon completion of the
fluorination, we carried out water leaching, which allowed transferring the undecom-
posed fluoroammonium salts into the solution. The results of fluorination, given in Table 2, show that SiO2 content is significantly
reduced in the samples. At the same time, the size of material does not affect the
effectiveness of desilication. Table 2. Chemical composition of samples after fluorination
No. Class, mm Mass fraction, %
Al2O3 SiO2 Fe2O3 TiO2 MgO K2O
1
IA
0.25–0.5
0.125–0.25
80.77
81.17
7.75
7.73
6.75
6.78
3.60
3.67
0.33
0.32
0.25
0.23
2
Vsch
0.25–0.5
0.125–0.25
73.94
71.81
8.98
11.24
9.85
10.07
3.92
3.67
1.03
1.00
1.66
1.58
3
V-V
0.25–0.5
0.125–0.25
55.07
52.87
7.28
9.17
30.20
30.61
2.38
2.29
2.72
2.74
1.40
1.36
4
Vsch
0.25–0.5
0.125–0.25
68.00
67.31
9.99
9.67
17.55
18.52
1.45
1.48
1.46
1.44
1.01
1.02
Admixture components Na2O, CaO, MnO, P2O5 no more
than 0.9%. Table 2. Chemical composition of samples after fluorination 3
Results and Discussion X-ray fluorescence analysis showed that the granulometric differentiation did not lead
to a change in the chemical composition and silicon module of the samples (Table 1). We diagnosed reflexes of kaolinite with hematite admixture on diffraction patterns of
samples No. 1, 2, and 3, and boehmite, kaolinite with anatase admixture on the patterns
of sample No. 4. Table 1. Chemical composition of off-grade bauxite ores
No. Class, mm Mass fraction, %
MSi
Al2O3 SiO2
Fe2O3 TiO2 MgO K2O
1
IA
0.25–0.5
0.125–0.25
43,82
44,08
50,83
50,76
2,93
2,70
1,66
1,66
0,31
0,31
0,10
0,10
0.86
0.86
2
Vsch
0.25–0.5
0.125–0.25
43,32
43,63
48,40
48,67
4,96
3,92
1,59
1,78
0,67
0,79
0,67
0,71
0.88
0.90
3
V-V
0.25–0.5
0.125–0.25
35,94
36,01
39,83
39,99
19,84
19,75
1,37
1,27
1,76
1,77
0,68
0,65
0.90
0.90
4
Vsch
0.25–0.5
0.125–0.25
59,85
59,37
23,91
23,78
12,98
13,53
1,12
1,15
1,03
1,04
0,61
0,62
2.50
2.50
Admixture components Na2O, CaO, MnO, P2O5 no more than 0.8%. Table 1. Chemical composition of off-grade bauxite ores Thermodynamic calculations of fluorination of kaolinite NH4HF2 are given else-
where (Rimkevich et al. 2016). However, the theoretical equation turns out to be
complicated in stoichiometric terms, which leads to the overconsumption of the
fluorination component. Our works on fluorination of titanium ores of the Yarega
deposit in the Komi Republic showed the efficiency of applying the ratio calculation to
the target component—silicon oxide (Perovskiy and Ignat’ev 2013; Perovskiy and
Burtsev 2016). SiO2 fluorination can be described by the following equation: Application of Fluoride Technology for Processing 43 SiO2 þ 3NH4HF2 ¼
NH4
ð
Þ2SiF6 þ 2H2O þ NH3 " 4
Conclusions The process of hydrofluoride desilication has been applied to off-grade aluminum raw
materials widespread in the Komi Republic. The optimal ratio of NH4HF2: SiO2 equal to
1:1 was determined by DTA method. At the fluorination temperature 300 °C and timing
30 min, concentrates with Al2O3 content more than 70–80%, and SiO2 - less than 10%
were produced. This technology is an alternative to the acid and alkaline processing of
high-siliceous aluminum raw materials. The use of fluoride technology allows not only
to improve the quality of ores because of desilication, but also to obtain products that can
be directly processed by electrothermal methods to silumin (Lepezin et al. 2014), fer-
roalloys (Bukin and Seregin 2014) and other products with high added value. Acknowledgements. The work was carried out with the financial support of the State task
“Scientific basis for effective subsoil use, development and exploration of mineral resource base, I. Burtsev et al. 44 development and implementation of innovating technologies and economic zoning of the Timan-
Nothern Ural region” No. AAAA- A17-117121270037-4. development and implementation of innovating technologies and economic zoning of the Timan-
Nothern Ural region” No. AAAA- A17-117121270037-4. Institute of Comprehensive Exploitation of Mineral Resources,
Russian Academy of Sciences, Moscow, Russia
vchan@mail.ru Abstract. The main methodological parameters and the modes of investigation
of samples containing micro- and nano-sized reagent phases are determined. Using scanning laser microscopy and an original method for analyzing the
surface relief, a quantitative assessment of the adsorption layer of the collector
reagent on Au-sulfide minerals was performed, the proportion of the molecular
form of adsorption and the retention agent fixing strength was calculated. The
theoretical basis for choosing the reagent mode for selective flotation of mul-
ticomponent ores was developed. The action mechanism has been revealed and
the prospect of using novel selective collectors and environmentally friendly
plant reagents for extracting non-ferrous and noble metals from complex sulfide
ores has been substantiated. Keywords: Reagent Adsorption Nanoparticles Mineral Microscopy
Flotation Keywords: Reagent Adsorption Nanoparticles Mineral Microscopy
Flotation © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 45–48, 2019.
https://doi.org/10.1007/978-3-030-22974-0_11 References Bukin AV, Seregin AN (2014) Development of technology for smelting ferrosilicoaluminium
from sub-standart bauxite and aluminous waste of metallurgy and electrical power energetic. Probl Ferr Metall Mater Sci (2):31–36 Burtsev IN, Kotova OB, Kuzmin DV, Mashin DO et al (2016) Prototypes of new technologies
for development of the mineral raw materials complex of the timan-north urals region. In:
Proceedings of the Komi science centre of the Ural division of the Russian academy of
sciences, vol 27, no 3, pp 79–88 Dyachenko AN, Kraidenko RI (2007) Separation of silicon-iron-copper-nickel concentrate by
fluorammonium method into individual oxides. Bull Tomsk Polytech Univ 311(3):35–38 Lepezin GG, An’shakov AS, Faleev VA, Avvakumov EG, Vinokurova OB (2014) Plasma-
chemical method of producing silumin and aluminium from the minerals of the sillimanite
group. Doklady Chem 456(2):110–113 Medkov MA, Krysenko GF, Epov DG (2011) Ammonium bifluoride – the perspective reagent
for complex processing of mineral raw materials. Vestn Far East Branch Russ Acad Sci. 159
(5):60–65 Perovskiy IA, Burtsev IN (2016) Influence of mechanical activation of leucoxene on efficiency of
its processing by fluoride method. Perspektivnye Materialy (2):66–73 Perovskiy IA, Ignat’ev GV (2013) Ammonium fluoride method of desilication of leucoxene
concentrate of Yarega deposit. In: Predictive estimate of technological properties of minerals
by applied mineralogy methods. Proceedings of 7th Russian seminar of process mineralogy,
Karelian Scientific Centre RAS, Petrozavodsk, pp 110–116 Rimkevich VS, Pushkin AA, Girenko IV, Leontyev MA (2016) Perspectives of complex
processing the kaolin concentrates by hydrochemical method. Izv Samara Sci Cent Russ Acad
Sci 18(2):186–190 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Applied Mineralogy for Complex
and Profound Mineral Processing V. Chanturiya(&) and T. Matveeva Institute of Comprehensive Exploitation of Mineral Resources,
Russian Academy of Sciences, Moscow, Russia
vchan@mail.ru Institute of Comprehensive Exploitation of Mineral Resources,
Russian Academy of Sciences, Moscow, Russia
vchan@mail.ru 1
Introduction At present, modern mining and ore dressing plants have been facing a number of serious
challenges in processing low-grade complex ores and technogenic resources, increasing
demands of high quality metal concentrates and ecologically safe methods of benefi-
ciation. In those conditions, the tasks of making mineral processing more complete and
comprehensive and of creating highly effective technologies come to the fore. These
should be based on the intensification of the existing methods and on creating new
methods of mineral extraction from hard-to-enrich ores and from technogenic deposits
by using the newest achievements of the fundamental sciences. The transition to the new
strategy of primary processing is only possible on technological-mineralogical evalu-
ation of raw minerals. A modern complex of high-resolution physical methods allows to investigate the
composition, structure and properties of geomaterials at the micro and nano-level,
including: • Identify micro and nano-sized particles of noble metals and surface natural and
artificial nano-formations on minerals; • Identify micro and nano-sized particles of noble metals and surface natural and
artificial nano-formations on minerals; • Experimentally substantiate the structural, phase and chemical transformations of
minerals under various energy methods of influence; • Experimentally substantiate the structural, phase and chemical transformations of
minerals under various energy methods of influence; © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 45–48, 2019. https://doi.org/10.1007/978-3-030-22974-0_11 © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 45–48, 2019. https://doi.org/10.1007/978-3-030-22974-0_11 V. Chanturiya and T. Matveeva 46 • To substantiate the choice and mechanism of interaction of reagents with noble
metals during flotation of complex ores of complex material composition; • To substantiate the choice and mechanism of interaction of reagents with noble
metals during flotation of complex ores of complex material composition; • Investigate the structural, phase and chemical transformations of sulfides and rocks
in heap leaching processes. • Investigate the structural, phase and chemical transformations of sulfides and rocks
in heap leaching processes. 2
Methods and Approaches Research methods are optical, confocal laser, analytical electronic, scanning probe
microscopy, UV-spectrophotometry of reagent solutions, flotation of minerals. The KEYENCE scanning laser microscope with the surface analysis module VK-9700
enables making a non-contact measurement of the roughness of the surface of minerals
and thus determining the height and size of the new formations obtained as a result of
interaction with the reagents. The electronic microscope with energy-dispersive
microanalyzer LEO-1420 VP INCA-350 allows determining the elemental composition
of micro- and nanophases of reagents on the surface of minerals. The analysis of the
surface of minerals before and after a contact with reagent solutions was carried out on
polished sections made in the form of polished plates 10 10 2 mm in size. 3
Results and Discussion In IPKON RAS within the framework of the scientific school of academician V. A. Chanturiya a complex of theoretical and experimental studies on the research and
testing of new classes of complex-forming reagents – collectors and modifiers for
flotation extraction of non-ferrous and noble metals from refractory gold-bearing
oreshave been made. To analyze the processes of physicochemical effects of flotation
reagents on micro and nano inclusions of gold and platinum on the surface of sulfides,
the authors first developed and improved methods for obtaining samples of mineral
complexes that mimic natural sulfides containing “invisible” or submicron gold or
platinum. The new highly effective reagents developed in IPKON RAS are modified
diethyldithiocarbamate DEDTC and diisobutyldithiophosphinate DIFM, dithiazine
derivatives MTX and dialkylpyrylmethane DAM. Those reagents showed the effect for
extracting micro- and nanoparticles of noble metals while enriching the mineral raw
materials of complex composition owing to formation of poorly water soluble gold
compounds and their selective adsorption on gold containing sulfides, improving the
flotation selectivity compared to traditional xanthate collector (Chanturiya 2017;
Chanturiya et al. 2016; Matveeva et al. 2017a; Matveeva et al. 2017b). Studies were
carried out using a set of UV and IR spectroscopy methods (UV-1700 Shimadzu and
Infralum FT-8), analytical scanning electron (ASEM) (LEO 1420VP with INCA
Oxford micro 350) and laser (KEYENCE VK-9700) microscopy, X-ray phase analysis
(X-ray diffractometry). The results made it possible to establish the conditions for the
formation of an adsorption layer of new reagents on micro- and nanoparticles of noble
metals and to ensure an increase in gold recovery during flotation (Fig. 1). Applied Mineralogy for Complex and Profound Mineral Processing 47 keV
Fig. 1. Adsorbed layer of collector DEDTCm on the surface of Au-pyrite. Fig. 1. Adsorbed layer of collector DEDTCm on the surface of Au-pyrite. As the major losses, from 25% to 30% of valuable components (gold and minerals
of the platinum group), are accounted for by the micro- and nano-size of the mineral
particles whose concentration in the ore does not generally exceed 1,5–3,0 g/t, it was
necessary to obtain samples imitating natural minerals. IPKON RAS has scientifically
substantiated and offered procedures for artificially coating minerals with Au and Pt
micro- and nano-particles, which made it possible to investigate the interaction
mechanism for a new class of frothing agents with noble metals. 3
Results and Discussion The introduction of the new agents both increased the extraction and improved the
quality of the mineral concentrate in the mineral processing of rebellious ores of noble
metals with a complex material composition, Table 1. Table 1. Indicators of flotation of the low-grade sulfide ore of the Fyodorovo-Panskoye deposit
when employing DEDTCm and ButX
Flotation products bPd
bPt
ɛPd
ɛPt
ButX Conc. 3.88 1.02 73.97 72.33
Tails
0.21 0.06 26.03 27.67
Ore
0.7
0.19 100
100
DEDTCmConc. 10.14 4.24 82.23 85.31
Tails
10.18 0.06 17.77 14.69
Ore
0.93 0.38 100
100 le 1. Indicators of flotation of the low-grade sulfide ore of the Fyodorovo-Panskoye deposit
n employing DEDTCm and ButX The implementation of new technologies at the mining-enrichment works in Russia
will make it possible to increase metal extraction by 10–15%, to obtain high-grade
finished products that are competitive in the world market, to involve unpayable ores
and technogenic raw materials into processing and considerably ameliorate negative
effects on the environment in the mining industry regions. V. Chanturiya and T. Matveeva 48 4
Conclusions The development of the mining sciences should be based on modern achievements of
applied mineralogy and innovative development of comprehensive, economic and
effective exploitation of mineral resources. The novel methods ensure both the higher
level of mineral extraction, the higher grade of obtained valuable components and high
level of ecological safety. Acknowledgements. The authors are grateful to the Grant of the President of RF “Scientific
School” of acad. V.A. Chanturiya and Program of Presidium of RAS P39. Chanturiya VA, Matveeva TN, Ivanova TA, Getman VV (2016) Mechanism of interaction of
cloud point polymers with platinum and gold in flotation of finely disseminated precious
metal ores. Miner Process Extr Metall Rev 37(3):187–195 Chanturiya VA (2017) Scientific substantiation and development innovative complex processing
of mineral resources. Gornyi J 11:7–13 References Chanturiya VA (2017) Scientific substantiation and development innovative complex processing
of mineral resources. Gornyi J 11:7–13 Chanturiya VA, Matveeva TN, Ivanova TA, Getman VV (2016) Mechanism of interaction of
cloud point polymers with platinum and gold in flotation of finely disseminated precious
metal ores. Miner Process Extr Metall Rev 37(3):187–195 Matveeva TN, Gromova NK, Minaev VA, Lantsiova LB (2017a) Modification of sulfide
minerals surface and cassiterite by stable complexes Me–dibutylditiocarbamate. Obogashche-
nierud 5:15–20. https://doi.org/10.17580/or.2017.05.03 Matveeva TN, Ivanova TA, Getman VV, Gromova NK (2017b) New flotation reagents for
extraction of micro- and nano-particles of noble metals from refractory ores. Gornyi J 11:89–93 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Mineralogical and Technological Features
of Tin Minerals at Pravourmiysky Deposit
(Khabarovsk Region) T. Chikisheva1,2,3(&), S. Prokopyev1,2,3, E. Kolesov4, V. Kilin4,
A. Karpova1,3, E. Prokopyev1,2, and V. Tukuser1,3 1 LCC PC «Spirit», Irkutsk, Russia
chikishevatatyana@mail.ru
2 Institute of the Earth Crust SB RAS, Irkutsk, Russia
3 Irkutsk State University, Irkutsk, Russia
4 PJSC «RUSOLOVO», Moscow, Russia 1 LCC PC «Spirit», Irkutsk, Russia
chikishevatatyana@mail.ru
2 Institute of the Earth Crust SB RAS, Irkutsk, Russia
3 Irkutsk State University, Irkutsk, Russia
4 PJSC «RUSOLOVO», Moscow, Russia Abstract. The paper presents the data obtained in the process of mineralogical
studies of technological samples of tin ore from the Pravourmiysky deposit. The
authors studied in detail the mineralogical features of ores and tin-containing
minerals and their significance for the enrichment technology. During of the
study, the information on the main technological properties of the ore of the
deposit was clarified and supplemented. As a result of the study of the mineral
composition, the mineralogical features of the ore were identified, allowing to
select the methods of ore enrichment and predict the quality of the concentrates
and products obtained. The causes of loss of tin with tailings were established. The obtained data on the mineral composition, properties of ore minerals, and
textural and structural features of the ores will be applied when modernizing
their enrichment technology at the processing plant. Keywords: Mineralogical research Tin ores Tin minerals Keywords: Mineralogical research Tin ores Tin minerals © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 49–52, 2019.
https://doi.org/10.1007/978-3-030-22974-0_12 2
Methods and Approaches The object of the research was technological samples of tin ore from the Pravourmiisky
deposit. The mineral composition of the ore and the quantitative assessment of the
contents of each mineral were determined using the methods of optical microscopy and
using x-ray methods. Mineralogical analysis of ore crushed to a particle size of less
than 2 mm was carried out according to the methods of the Scientific Council on
methods of mineralogical research. The sequence of operations to determine the
mineral composition of the ore samples consisted in dividing the initial material of the
samples into size classes, followed by gravitational fractionation and studying the
distribution of minerals into fractions for each size class. 1
Introduction In the 21st century, tin is very in demand in the global economy due to its use in new
industries, the introduction of innovative technologies and the environmental friend-
liness of metal. At the same time, the tin market is volatile, depending on sharp
fluctuations in the price situation and annual changes between supply and demand [1]. Russian industry consumes about 6.5–7 thousand tons of tin per year. About 90% of
the mined tin is imported. Tin deposits in Russia are among the richest in the world. Pravourmiyskoe tin deposit is one of the promising ore deposits of tin. In addition,
together with tin, tungsten can be mined from ore deposits. To achieve a more complete
extraction of valuable components from the ore, a detailed study of the mineral com-
position of ores, the textural and structural features of the ore, the physical properties of
minerals and the degree of their contrast is necessary. Mineralogical analysis is the basis
for the study of the material composition, structure and texture, choice of directions and
methods of preparing raw materials for processing, enrichment technologies and © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 49–52, 2019. https://doi.org/10.1007/978-3-030-22974-0_12 T. Chikisheva et al. 50 metallurgy [2]. the paper is devoted to the study of the mineralogical and technological
features of the Pravourmiysky deposit tin ores in the Far East. 3
Results and Discussion The main ore mineral of tin at the Pravourmiysky deposit is cassiterite. Stannin,
mawsonite, stannoidite are present in small quantities. More than 98% of the tin in the
ore is in cassiterite, 1–2% of the tin is in the sulphide minerals of tin. Streaks, inclusions and clusters of grains of cassiterite, arsenopyrite and lellingite
represent ore mineralization. The size of individual grains of ore minerals is up to
5.0 mm, the apparent thickness of clusters is up to 60 mm. Cassiterite is observed in
the form of xenomorphic grains, their clusters; crystals of a prismatic, pyramidal-
prismatic appearance are less often observed (Fig. 1). The grains size of cassiterite
varies from 0.01 to 2.0 mm, with a predominance in the range of 0.1–0.5 mm, the
thickness of clusters of 3–5 mm, sometimes can reach up to 10.0 mm. Fig. 1. Micrograph of cassiterite (1) in the intergrowth with arsenopyrite (2), lellingite (3). Fig. 1. Micrograph of cassiterite (1) in the intergrowth with arsenopyrite (2), lellingite (3). According to the mineralogical analysis, crushed ore mainly consists of fragments
of greisen, quartz, topaz, tourmaline, feldspar and mica. Ore minerals (arsenopyrite, Mineralogical and Technological Features of Tin Minerals 51 lellingite, bornite and chalcopyrite) in crushed ore are in the form of fragments of
crystals and grains of irregular shape. Sometimes they are found in intergrowths with
each other and with rock-forming minerals. Cassiterite is observed in the form of irregular shape grains, as well as in inter-
growths with rock-forming minerals, less often with arsenopyrite or lellingite. The
results of the study of the disclosure of cassiterite grains are shown in Table 1. Table 1. Disclosure of cassiterite grains in crushed ore
Size class, mm
Cassiterite grains and cassiterite –
rich intergrowth, %
Cassiterite-containing
intergrowth, %
Total
−2+1
96,9
3,1
100,0
−1+0,5
98,9
1,1
100,0
−0,5+0,315
98,2
1,8
100,0
−0,315 +0,25
99,8
0,2
100,0
−0,25+0,125
99,8
0,2
100,0
−0,125+0,071
99,9
0,1
100,0
−0,071 +0,040
100,0
0,0
100,0
−0,040+0,0
100,0
0,0
100,0 Table 1. Disclosure of cassiterite grains in crushed ore According to Table 1, in ore crushed to a particle size of less than 2 mm, cassiterite
grains are mainly in the form of free grains. The maximum number of intergrowths
(3.1%) is noted in the size class −2 + 1 mm. The complete disengagement of cassi-
terite grains from intergrowths is achieved in size less than 0.071 mm. 3
Results and Discussion Tin sulphide minerals (stannin, mawsonite, stannoidite) are associated with later
copper mineralization. These minerals are in close intergrowth with chalcopyrite and
bornite. Their size usually does not exceed 0.1 mm. T. Chikisheva et al. Studying the contrast of the physical properties of ore minerals provided an idea of
the mineral composition of the products of primary gravity enrichment, which will
make it possible to develop an optimal scheme for finishing operations. Acknowledgements. The Irkutsk State University, individual research grant №091-18-231,
supported this work. Acknowledgements. The Irkutsk State University, individual research grant №091-18-231,
supported this work. supported this work. 1. Bashlykova TV (2005) Tekhnologicheskie aspekty racional’nogo nedropol’zovaniya: Rol’
tekhnologicheskoj ocenki v razvitii i upravlenii mineral’no-syr’evoj bazoj strany [Techno-
logical aspects of rational subsoil use: The role of technological assessment in the development
and management of the country’s mineral resource base]. Moscow^ MiSIS (in Russian) 4
Conclusions Tin minerals, which are valuable components of the studied samples, have a signifi-
cantly higher degree of contrast of gravitational properties, which can be used for their
primary concentration Together with cassiterite and stannine, wolframite, arsenopyrite
with lellingite and copper sulphides will be extracted into the primary gravity con-
centrate. To obtain tin concentrate that meets the requirements for raw materials,
arsenopyrite and copper sulfides can be extracted from the rough concentrate using
flotation methods. During the flotation process, the sulfide minerals of tin (stannin, mawsonite,
stannoidite) are extracted into sulfide products along with the minerals of copper and
arsenic. To extract tin sulfides minerals, selective flotation with preliminary fine
grinding of sulfide products will be required. Differences in the magnetic susceptibility of minerals, with the use of magnetic
separation, wolframite, biotite and tourmaline can be distinguished. 52 2. Danilov UG, Grigoryev VP (2017) Problemy i perspektivy razvitiya olovyannoj promysh-
lennosti Rossii. Gornaya promyshlennost’ 5(135):83–87 (in Russian) © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 53–58, 2019.
https://doi.org/10.1007/978-3-030-22974-0_13 Ore Mineralogy of High Sulfidation Çorak-Taç
Epimesothermal Gold Deposit
(Yusufeli-Artvin-Turkey) K. Diarra1, E. Sangu1, and E. Çiftçi2(&) 1 Department of Geological Engineering, Faculty of Engineering,
KOU, 41380 Kocaeli, Turkey
2 Department of Geological Engineering, Faculty of Mines,
ITU, 33469 Maslak, Istanbul, Turkey
eciftci@itu.edu.tr Abstract. Çorak and Taç, two nearby mineralizations, are located in the eastern
black sea region, which is one of the most productive metallogenic belts of Turkey. It is characterized by a great number of Kuroko-type volcanogenic massive sulfide
deposits as well as vein-type polymetallic deposits, porphyry and epithermal
precious metal deposits. Subject neighboring deposits are hosted within the
voluminous Cretaceous-Eocene granitoids and interbedded volcanic rocks and
carbonates. Mineralogy of altered host rocks include quartz veins, carbonates,
sericite, chlorite, chalcedony, and disseminated sulfides - mainly pyrite, sphalerite,
galena, and chalcopyrite. The main texture encountered in the host rocks is hyalo-
porphyry. Due to hydrothermal alterations primary minerals are mostly altered in
which the ferromagnesian minerals are chloritized and calcified, while feldspars
are altered into sericite, calcite, and albite. Silicification and argillic alteration
(medium, moderate, high) are widely spread however; XRD analysis carried on 33
core samples from Çorak has also revealed local propylitic alteration, limoniti-
zation and hematitization as well. The minerals assemblages that accompanied the
different alterations include jarosite and alunite suggesting high sulfidation
hydrothermal mineralization. Through the ore microscopic studies, pyrite, chal-
copyrite galena, sphalerite, and a lesser amount of sulfosalts (tennantite-
tetrahedrite and pyrargyrite) were determined. Quartz and calcite account for
the main gangue minerals. While the Taç mineralization is pyrite, chalcopyrite and
sphalerite dominating, the Çorak mineralization contains relatively less chal-
copyrite and galena becomes prevalent with sphalerite. Gold in both sites may
reach up to 10 ppm, on average 3 ppm. Silver occurrence is insignificant. Keywords: Yusufeli Artvin Turkey High sulfidation Gold
Epimesotherma References 1. Bashlykova TV (2005) Tekhnologicheskie aspekty racional’nogo nedropol’zovaniya: Rol’
tekhnologicheskoj ocenki v razvitii i upravlenii mineral’no-syr’evoj bazoj strany [Techno-
logical aspects of rational subsoil use: The role of technological assessment in the development
and management of the country’s mineral resource base]. Moscow^ MiSIS (in Russian) 2. Danilov UG, Grigoryev VP (2017) Problemy i perspektivy razvitiya olovyannoj promysh-
lennosti Rossii. Gornaya promyshlennost’ 5(135):83–87 (in Russian) 2. Danilov UG, Grigoryev VP (2017) Problemy i perspektivy razvitiya olovyannoj promysh-
lennosti Rossii. Gornaya promyshlennost’ 5(135):83–87 (in Russian) Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Keywords: Yusufeli Artvin Turkey High sulfidation Gold
Epimesotherma K. Diarra et al. Taç and Çorak deposits occur in an area about 10 km 3 km in size, oriented
NW-SE and SW-NW, respectively. It is limited to the north by series of outcrop of
large unaltered basaltic basement with some andesite enclave. They are covered by
more recent tuffs, sandstone and old alluvial fan. The south is more dominated by
andesites and tuff outcrops together with relatively recent sedimentary covers. Mineralization and alteration are represented by extensive strata-bound argillic
alteration cut by gold and base metal bearing quartz veins within a massive pyroclastic
volcanic host-rock below a bedded volcanogenic succession, with additional receptive
massive volcanic units occurring higher up in the sedimentary succession. Porosity is
believed to be the most likely control on the stratiform nature of alteration. However,
proximity to major structures along or close to the structural axis appears to be an
important influence on the presence of alteration suggesting structural control on
mineralization. Some typical structures are associated with the Cu-Pb-Zn mineralization: The most
obvious one is the hydrothermal alteration with its colored intense argillic and
propylitic alterations. They are always associated with a local fault. Afterward, the
large intrusive rocks are crosscutting the volcanic, the pyroclastic and the volcano
sedimentary rocks. In Çorak SW-NE section view, the high grade of base metal representing the
mineralization is localized in volcano sedimentary rock (tuff) as well as in andesite and
feldspar porphyry andesitic, the three sets are separated by set of faults. 1
Introduction The study area is located in Yusufeli area in the eastern Pontides’ northeastern most tip
within a larger metal rich tectonic corridor stretching from southern Georgia and
northern Armenia to Bulgaria and Romania and it is a host to numerous volcanogenic
massive sulfide (VMS) and vein-type deposits dominantly of Late Cretaceous age. 54 K. Diarra et al. 3
Results and Discussion The andesite and andesitic tuffs are the main lithologies hosting the mineralization in
the study area. The mineralogy of the altered hostrock enclosed quartz veins, carbonate,
sericite, chlorite, and chalcedony and disseminated sulfides. Their details microscopic
studies have revealed the following common textures: trachytic; hyalo microlithic
porphyry; vesicular microlithic porphyry; hyalo micro granular porphyry and quasi-
oolithic. The hydrothermal alteration manifest itself in the form of silicification, epi-
dotization, chloritization, seritization, and finally a calcification as a last stage. Colored
argillic alteration is widespread in the area along with silicic and propylitic alteration. Drill holes intercept sulfide ore zones at different depth. The sulfide mineralizations
may range from 50 cm to 1 m in thickness. Weakly mineralized zones of 1 to 2% with
disseminated sulfides are intercalated between the massive or semi massive sulfide
lenses. The altered interval shows 3 types of minerals assemblages: Quartz
+ sericite + calcite, quartz + epidote and quartz + chlorite + calcite: (I) quartz +
chlorite alteration: In this facies, the ferromagnesian (Amphibole) are completely
chloritized and calcified. Chlorites are filling cavity as well. They wrap also the fewer
amount of chalcedony present in the groundmass. The opaque minerals represent less
than 1–2% of the groundmass. The texture is vesicular hyalo-microlithic porphyry;
(II) quartz + epidote alteration: In this facies, the light colored plagioclases are
turned into epidote. The calcite as a last phase is filling fracture. (propylitic alteration);
(III) quartz + sericite alteration: The sericite is not only replacing the ferromagne-
sian and the plagioclase but also the matrix. Plagioclases turned into sericite. Euhedral
quartz is present in the groundmass. The most prevalent ore mineral in Çorak is pyrite it
is present in all the samples. It is followed by chalcopyrite, sphalerite, and galena. Compare to sulfides, sulfosalts namely tennantite-tetrahedrite are present in lesser
amounts. Covellite and bornite are rare. Gangue minerals are mainly composed of
quartz and calcite. Two main mineral’s assemblages are prevailing whether it is from
stockwork (disseminate and veinlet zone) or from the high grade sulfide zone. In the
stockwork zone pyrite has euhedral to subhedral shape. Chalcopyrite is anhedral and
replaces very often pyrite. In the massive ore zone, pyrite and chalcopyrite are
accompanied by large sphalerite and galena grain. Galena always crosscut sphalerite,
suggesting that it is later than sphalerite. Tennantite and tetrahedrite assemblage replace
very often pyrite and chalcopyrite in this zone. 2
Methods and Approaches For mineralogical and ore microscopy studies, 35 thin and polish sections from core
drill and outcrop, have been prepared and scrutinized under reflect and transmitted light
microscope of the laboratories of Kocaeli university (KOÜ) and Istanbul technical
university (ITU) in Turkey. 6 samples have been taken from alteration zone have been
analysis in order to compute their CIA: ([Al2O3/(Al2O3 + CaO + K2O + Na2O)]
100) and ICV: (ICV: CaO + K2O + Na2O + Fe2O3 (t) + MgO + MnO + TiO2)/
Al2O3). The target was to determine their depositional environment as well as the
alteration styles. 33 samples from different alteration zones from Çorak have been
submitted to the lithological and XRD analysis using Rietveld methods. g
y
g
15 polished thin sections have been subjected to Cathodoluminescence (CL) mi-
croscopy study to describe and to distinguish gangue minerals and minerals genera-
tions. Samples containing mostly sphalerite, quartz and carbonates were chosen, since
they are able to produce CL emissions. The CL study was performed at the ITU
Advanced Microscope lab using an optical cathodoluminescence system (CITL MK5
system). 15 polished thin sections have been subjected to Cathodoluminescence (CL) mi-
croscopy study to describe and to distinguish gangue minerals and minerals genera-
tions. Samples containing mostly sphalerite, quartz and carbonates were chosen, since
they are able to produce CL emissions. The CL study was performed at the ITU
Advanced Microscope lab using an optical cathodoluminescence system (CITL MK5
system). Ore Mineralogy of High Sulfidation Çorak-Taç Epimesothermal Gold Deposit Ore Mineralogy of High Sulfidation Çorak-Taç Epimesothermal Gold Deposit Ore Mineralogy of High Sulfidation Çorak-Taç Epimesothermal Gold Deposit 55 55 3
Results and Discussion The pyrargyrite is present in very small
amount. The tennantite is dominant in Çorak’s polymetallic sulfide zone. It has been
also observed galena replacing gangue minerals quartz suggesting symplectic inter-
growths of ore minerals with silicate. Colloform texture is observed but they are rare. Except the pyrite, all the minerals are anhedral. Minerals size span from 50 to 300
micron. The sulfosalts are the smallest ones (Figs. 1a–f). K. Diarra et al. 56 Fig. 1. (a) galena replacing chalcopyrite; (b) galena replacing quartz, replacement of gan
minerals by ore minerals (symplectic intergrowths) and tennantite replacing chalcopyrit
(c) galena veins sphalerite, while pyrite replaced by tennantite; (d) tennantite replacing pyrite an
chalcopyrite; (e) tetrahedrite veining galena and tetrahedrite-tennantite replacing sphalerite (P
galena; Cp: chalcopyrite; Sph: sphalerite; Tn-Tt: Tennantite-Tetrahedrite; Qtz quartz) Fig. 1. (a) galena replacing chalcopyrite; (b) galena replacing quartz, replacement of gang
minerals by ore minerals (symplectic intergrowths) and tennantite replacing chalcopyrite;
(c) galena veins sphalerite, while pyrite replaced by tennantite; (d) tennantite replacing pyrite and
chalcopyrite; (e) tetrahedrite veining galena and tetrahedrite-tennantite replacing sphalerite (Py:
galena; Cp: chalcopyrite; Sph: sphalerite; Tn-Tt: Tennantite-Tetrahedrite; Qtz quartz) The following paragenesis is deducted from the ore microscopy study: Pyrite –
Chalcopyrite (I) – Sphalerite – Galena – Sulfosalts- Chalcopyrite (II) - Bornite –
Covellite. The following paragenesis is deducted from the ore microscopy study: Pyrite –
Chalcopyrite (I) – Sphalerite – Galena – Sulfosalts- Chalcopyrite (II) - Bornite –
Covellite. The cathodoluminescence study on sphalerite has revealed heterogeneity in the
composition of the sphalerite (Fig. 2). These states that those sphalerite grains are
originated from more than one generation phase and/or from fluids with changing
composition. Ore Mineralogy of High Sulfidation Çorak-Taç Epimesothermal Gold Deposit 57 Fig. 2. Optical cathodoluminescence microscopy images (OCLMI) of sphalerite and quartz
grains with infilling late calcite, showing compositionally heterogeneity sphalerites Fig. 2. Optical cathodoluminescence microscopy images (OCLMI) of sphalerite and quartz
grains with infilling late calcite, showing compositionally heterogeneity sphalerites 4
Conclusions Taç and Çorak deposits occur in north easternmost tip of the eastern Pontides tectonic
belt of Turkey. They are two separate ore bodies within the same system. Hydrothermal
alteration is intense and extensive in the form of silicification, epidotization, chloriti-
zation, seritization, and finally a calcification as the final stage. Three main alterations
zone are succeeding from the proximal, intermediate to distal zone: Quartz + Chlorite
alteration, quartz + epidote alteration, quartz + sericite alteration. Two main mineral’s
assemblages are prevailing whether it is from stockwork (disseminated and veinlet) or
from the high grade sulfide zone. They are respectively: pervasive pyrite, chalcopyrite
with sphalerite and galena. Argillic, propylitic and silicic alteration induced by acidic hydrothermal fluids have
led to the formation of quartz, muscovite, orthoclase, gypsum, dolomite, kaolinite,
pyrite, sphalerite, galena and the pair jarosite-alunite assemblage. The presence of
alunite and jarosite minerals indicate that Çorak and Taç are high sulfidation epithermal
deposit with mesothermal signatures. Acknowledgements. This study was financially supported by BAP Project unit (No: 2017/80)
of Kocaeli University in Turkey. We are thankful to the university and to Santral Mining Co. and
their employees in Yusufeli for their courteous support for the field work. 1. Bogdanov B (1980) Massive sulphide and porphyry copper deposits in the Panagjurishte
district, Bulgaria. In: Jankovic S, Sillitoe Richard H (eds) European copper deposits;
Proceedings of an international symposium. Springer, Berlin-Heidelberg-New York, pp 50–58 2. Ciftci E (2000) Mineralogy, paragenetic sequence, geochemistry, and genesis of the gold and
silver bearing upper cretaceous mineral deposits, Northeastern Turkey. PhD dissertation,
University of Missouri-Rolla, Missouri, USA, p 251 3. Kouzmanov K, Moritz R (2009) Late Cretaceous Porphyry Cu and epithermal Cu-Au
association in the Southern Panagyurishte District, Bulgaria: The paired Vlaykov Vruh and
Elshitsa deposits 4. Muntean JL, Einaudi MT (2000) Porphyry gold deposits of the Refugio district, Maricunga
belt, Northern Chile. Econ Geol 95:1445–1472 Mineralogical and Technological Aspects
of Phosphate Ore Processing A. Elbendari(&), V. Potemkin, T. Aleksandrova, and N. Nikolaeva Mineral Processing Department, Saint Petersburg Mining University,
Saint Petersburg, Russia
abdullah_elbendary@yahoo.com Abstract. The article studies the mineralogical features of phosphate ores. In
the conditions of declining industrial reserves of apatite-containing ores, issues
of a more comprehensive and in-depth study of the mineral and material
composition, as well as the improvement of existing technologies for the pro-
cessing of this type of raw material, become topical. Using optical methods of
analysis, electron microscopy with automated mineralogical analysis (MLA),
mineral and elemental composition of apatite was obtained. Taking into account
the studied mineralogical and material composition, experiments on grinding
and flotation were carried out. Based on these data, it was concluded that the
optimal scheme for the processing of phosphate ores is a flotation scheme with
preliminary selective disintegration. Keywords: Mineralogical composition Grinding Beneficiation
Phosphorus-bearing minerals Keywords: Mineralogical composition Grinding Beneficiation
Phosphorus-bearing minerals © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 59–65, 2019.
https://doi.org/10.1007/978-3-030-22974-0_14 References 1. Bogdanov B (1980) Massive sulphide and porphyry copper deposits in the Panagjurishte
district, Bulgaria. In: Jankovic S, Sillitoe Richard H (eds) European copper deposits;
Proceedings of an international symposium. Springer, Berlin-Heidelberg-New York, pp 50–58 2. Ciftci E (2000) Mineralogy, paragenetic sequence, geochemistry, and genesis of the gold and
silver bearing upper cretaceous mineral deposits, Northeastern Turkey. PhD dissertation,
University of Missouri-Rolla, Missouri, USA, p 251 K. Diarra et al. 58 3. Kouzmanov K, Moritz R (2009) Late Cretaceous Porphyry Cu and epithermal Cu-Au
association in the Southern Panagyurishte District, Bulgaria: The paired Vlaykov Vruh and
Elshitsa deposits 4. Muntean JL, Einaudi MT (2000) Porphyry gold deposits of the Refugio district, Maricunga
belt, Northern Chile. Econ Geol 95:1445–1472 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches The object of the study was apatite-nepheline ore of the Khibiny deposits group
(Russia) and phosphate ore of the Abu-Tartur deposit (Egypt). For the development of
beneficiation schemes and modes, complex studies were carried out on mineral and
elemental composition, including optical methods of analysis, electron microscopy
using automated mineralogical analysis (MLA), etc. As a result of the work, the
mineral composition of apatite-nepheline ore (ANO) and phosphate ore (PO) was
studied taking into account the data of optical and electron microscopic studies, spot X-
ray spectral and chemical analyzes, atomic emission spectrometry, automated miner-
alogical analysis. The chemical composition of ANO and PO is given in the Table 1. Table 1. The chemical composition of ANO and PO
Apatite-nephelineores
(Russia)
Phosphoriteore (Egypt)
Component Content, % Component Content, %
SiO2
32.50
SiO2
19.4
TiO2
2.20
TiO2
0.06
Al2O3
15.52
Al2O3
2.6
Fe
5.45
Fe
5.3
CaO
18.28
CaO
37.6
MgO
0.92
MgO
0.8
MnO
0.16
MnO
0.05
K2O
4.20
K2O
0.59
Na2O
5.56
Na2O
0.57
P2O5
12.50
P2O5
21.8
SO3
0.04
SO3
1.6
CO2
0.05
CO2
1.2
LOI
2.62
LOI
8.43
Total
100.00
Total
100.00 Table 1. The chemical composition of ANO and PO To study the possibility of increasing efficiency of the flotation process, studies
were carried out on the selection of the optimal grinding mode and a series of flotation
experiments. 1
Introduction Phosphates are one of the most important minerals on Earth, as they are used as
fertilizers for agriculture and as a necessary raw material for the chemical industry
(Brylyakov 2004; Abouzeid 2007). In addition, phosphates are the source of rare-earth
elements. They are used in many commercial and industrial products, such as: deter-
gents, toothpastes and fireproof materials. Worldwide consumption of P2O5 in all of the
areas above is projected to grow gradually from 44.5 million tons in 2016 to 48.9
million tons in 2020 (Jasinski 2017). In the conditions of declining industrial high-quality reserves of phosphorus-
containing ores, issues of a more comprehensive and in-depth study of the mineral and
material composition, as well as improvement of existing technologies for processing
this type of raw material, become urgent. The study of the influence of the mineral raw
materials composition on the features of the beneficiation schemes construction is
given in the works of many authors (Aleksandrova et al. 2012; Evdokimova et al. 2012;
Gerasimova et al. 2018; Litvintsev et al. 2006; Mitrofanova et al. 2017). © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 59–65, 2019. https://doi.org/10.1007/978-3-030-22974-0_14 A. Elbendari et al. 60 3
Results and Discussion According to the mineralogical analysis, the main primary minerals of ANO are apatite
and nepheline, the contents of which are respectively 30.67 and 30.88%; minor
quantities contain pyroxenes, mica, feldspars, as well as natrolite and kaolinite –
secondary minerals formed due to the destruction of the primary mineral phases. Mineralogical and Technological Aspects of Phosphate Ore Processing 61 Phosphorus-containing minerals of the sample are apatite, eschynite, phosphates of
rare-earth elements and lomonosovite, with a distribution to these minerals of 99.94,
0.01, 0.02 and 0.03% phosphorus respectively. Valuable minerals of the sample are
apatite – the main mineral concentrating phosphorus and nepheline – the main mineral
concentrating aluminum. Apatite is the main mineral concentrator of phosphorus in the ore; it forms dis-
seminated prismatic crystals, vein clusters of crystals, less often massive clusters of
fractured xenomorphic grains, often included in the grains of other minerals - pyrox-
enes, mica, sphene, nepheline (Fig. 1). Fig. 1. Apatite crystals in the aegirine-augite matrix. Spectra in (b): 1, 7 - nepheline; 2, 4 -
aegirine-augit; 3, 8 - apatite; 5, 6 - arfvedsonit Fig. 1. Apatite crystals in the aegirine-augite matrix. Spectra in (b): 1, 7 - nepheline; 2, 4 -
aegirine-augit; 3, 8 - apatite; 5, 6 - arfvedsonit Image: (a) - in reflected light; (b) - in backscattered electrons. Apatite is charac-
terized by a pronounced idiomorphism of grains that have clear crystallographic out-
lines; the shape of apatite grains is columnar, prismatic, acicular, which causes a weak
connection between them in aggregates. The crystalline form of apatite, the natural
brittleness of the mineral will contribute to the primary destruction of the mineral
during ore grinding and the concentration of apatite in smaller grades. According to MLA data, 32.78% of the mineral in ore is distributed into free
particles, 22.29% into binary and 44.92% into polymineral intergrowth. Phosphate minerals in PO are represented by carbonate-fluoroapatite, fluoroapatite,
hydroxylapatite, and francolite. The most common clay minerals in the studied sedi-
ments are smectites. The amount of kaolinite and illite in general is insignificant,
although their content is also high enough. Among the non-phosphate components of
the PO, detrital quartz, as well as ankerite and pyrite cement in unaltered phosphates,
are predominant. Pyrite and ankerite in many cases replace partially or completely
phosphate grains. A. Elbendari et al. Based on the studies, it was found that this scheme is optimal for both samples
(sedimentary and volcanic). 4
Conclusions In the conditions of declining quality of industrial reserves of phosphate ores, issues of
a more comprehensive and in-depth study of the mineral and material composition, as
well as the improvement of existing technologies for the processing of this type of raw
material become topical. Achieving this goal is complicated by the constant decline in
the quality of ores involved in processing and requirement of 100% recycled water
supply implementation. Based on the mineralogical, chemical, material composition, as
well as technological research on the possibility of processing phosphate ores, it was
concluded that the optimal scheme for the extraction of apatite is a flotation circuit with
preliminary selective disintegration. At the same time, the possibility of extracting rare
earth metals has been established. Acknowledgements. The work is carried out under financial support of the Ministry of Edu-
cation and Science of the Russian Federation, the project RFMEFI57417X0168. 3
Results and Discussion Elements such as Ba, Cr, Ni, Sr, Y, and Zr are found in relatively
high concentrations, while Co, Nb, Pb, Rb, Th, and U are found in relatively low
concentrations (Abdel-Moghny and Zhabin 2011; Baioumy 2013) (Fig. 2). A. Elbendari et al. 62 Fig. 2. Electron microscopic images of PO (Baioumy 2013). (а) – rounded silica inclusions;
(b) – kaolinite plates Fig. 2. Electron microscopic images of PO (Baioumy 2013). (а) – rounded silica inclusions;
(b) – kaolinite plates To determine the optimal processing scheme for ANO and PO, samples with a
grain size of less than 2 mm and a mass of 550 g were ground in a ball mill in
accordance with the specifications given in Table 2: Table 2. Mill characteristics and experiment conditions
Mill
Inner diameter (D), mm
125
Length, mm
170
Volume, cm3
2085
Critical speed, rpm
120
Media (Balls)
Material
Alloy steel
d, mm
19
22
25
27
28
29
30
Number
1
2
2
4
4
3
1
d, mm
32
34
35
36
39
40
41
Number
2
1
2
1
1
2
1
Specific gravity
7.8
Mass of balls, g
3388.5
Material
Igneous & sedimentary phosphate ore
Specific gravity
3.1 & 2.7
Powder weight, g
550 gm Table 2. Mill characteristics and experiment conditions The grinding results are shown in Fig. 3. As a result of the grinding process study, it was found that with an increase in the
grinding time, the particle size sharply decreases, and pulp grinding with a solid content
of 50% gives the best results. Studies have also been conducted for the process of
grinding apatite ore with the addition of tributyl phosphate in amount of 500 and Mineralogical and Technological Aspects of Phosphate Ore Processing Mineralogical and Technological Aspects of Phosphate Ore Processing 63 Fig. 3. 3D relationship between the rotational speed and grinding time Fig. 3. 3D relationship between the rotational speed and grinding time 1000 ml/ton. Studies have shown that the addition of surface-active substances (tributyl
phosphate) during grinding of ANO and PO does not only increase the efficiency of
grinding, but also partially convert rare-earth metals into soluble form with their
subsequent extraction. Beneficiation of ANO and PO samples was carried out according to the flowchart
shown in Fig. 4. Fig. 4. Block diagram of phosphate ore beneficiation Fig. 4. Block diagram of phosphate ore beneficiation 64 A. Elbendari et al. Mitrofanova GV, Ivanova VA, Artemiev AV (2017) Use of reagents-flocculants in water-
preparation processes during phosphorous-containing ore processing. In: International
multidisciplinary scientific GeoConference surveying geology and mining ecology manage-
ment, SGEM 2017, vol 17, no 11, pp 1143–1150 Mineralogical and Technological Aspects of Phosphate Ore Processing © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 66–69, 2019.
https://doi.org/10.1007/978-3-030-22974-0_15 REC Mekhanobr-tekhnica, St-Petersburg, Russia
gerasimov_am@npk-mt.spb.ru REC Mekhanobr-tekhnica, St-Petersburg, Russia
gerasimov_am@npk-mt.spb.ru Abstract. The solid mineral raw materials of the sediment deposits containing
the argillaceous variations such as the coal and the potash ores are the most
multi-tonnage solid minerals mined and processed in the territory of the Russian
Federation. Both types of the deposits have relatively simple geology aspects
with large and medium-sized solids and as a rule, the most often, with the
regular bedding which are characterized by the fitchery thickness and the
internal structure. Such deposits are very difficult for mining and processing. The thermal modification of this of ores allows changing the physico-chemical
properties of its components in such a way that their further processing -
grinding, separation, transportation and storage can be carried out without the
use of process water or significantly reduces its consumption. Keywords: Argillaceous minerals Hydrochemical modification
Thermochemical modification Water saving Keywords: Argillaceous minerals Hydrochemical modification
Thermochemical modification Water saving References Abdel-Moghny MW, Zhabin AV (2011) Mineralogical features of phosphate rocks Egypt. Bull
Voronezh State Univ Ser Geol 2011(1):37–49 Abouzeid AM (2007) Upgrading of phosphate ores – a review. Powder Handling Process 19:92–
109 Aleksandrova TN, Litvinova NM, Gurman MA, Aleksandrov AV (2012) Comprehensive
utilization of the far eastern apatite-containing raw materials. J Min Sci 48(6):1047–1053 Baioumy HM (2013) Effect of the depositional environment on the compositional variations
among the phosphorite deposits in Egypt. Geol Geophys 54(4):589–600 Brylyakov YE (2004) The development of the theory and practice of complex enrichment of
apatite-nepheline ores of the Khibinsky deposits Evdokimova GA, Gershenkop ASh, Fokina NV (2012) The impact of bacteria of circulating
water on apatite-nepheline ore flotation. J Environ Sci Health - Part A Toxic/Hazardous Subst
Environ Eng 47(3):398–404 Gerasimova LG, Nikolaev AI, Maslova MV, Shchukina ES, Samburov GO, Yakovenchuk VN,
Ivanyuk GY (2018) Titanite ores of the khibiny apatite-nepheline-deposits: selective mining,
processing and application for titanosilicate synthesis. Minerals 8(10):446 Jasinski SM (2017) Phosphate Rock, U.S. Geological Survey, Mineral Commodity Summa Jasinski SM (2017) Phosphate Rock, U.S. Geological Survey, Mineral Commodity Summaries
Litvintsev VS, Melnikova TN, Yatlukova NG, Litvinova NM (2006) Mechanoactivation in the
processes of ore preparation. Mt J (6):95–96 Litvintsev VS, Melnikova TN, Yatlukova NG, Litvinova NM (2006) Mechanoactivation in the
processes of ore preparation. Mt J (6):95–96 Mineralogical and Technological Aspects of Phosphate Ore Processing 65 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Properties and Processing of Ores Containing
Layered Silicates A. Gerasimov(&), V. Arsentyev, and V. Lazareva
REC Mekhanobr-tekhnica, St-Petersburg, Russia
gerasimov_am@npk-mt.spb.ru A. Gerasimov(&), V. Arsentyev, and V. Lazareva REC Mekhanobr-tekhnica, St-Petersburg, Russia
gerasimov_am@npk-mt.spb.ru 2
Methods and Approaches Coal, sylvinite and kaolin were selected for the dry processing study. The presented coals are dark gray, dense, laminated and fractured with the matte
surface in the place of spalling. The samples of the coal are brittle and under a slight
stress are destroyed along the microfracture (Gerasimov et al. 2016). The sylvinite sample is a rock salt-sylvinite-carnalite formation with the varying
ratio of the main rock-forming minerals represented mainly by readily soluble con-
geries and individuals of the chloride class (Arsentyev et al. 2017a, b). The kaolin ore is represented by the kaolin argillites mainly gray, dark gray,
greenish-gray, less often yellowish-ochreous and sorrel, occasionally milky-gray colors
cured during the compaction, dehydration and cementation processes. The structure is
massive often the breccia structure and splintery, sometimes the oolitic structure, here
and there the indistinctly-banded. The structure is the pelitic, aleuropelitic, aleuro-
psammito-pelitic, occasionally oolitic (Arsentyev et al. 2017a, b). 1
Introduction Considering these minerals from the point Properties and Processing of Ores Containing Layered Silicates 67 of view of their advanced processing, it is important to note the similar processing
characteristics: of view of their advanced processing, it is important to note the similar processing
characteristics: of view of their advanced processing, it is important to note the similar processing
characteristics: – comparatively high content of the useful component which is 60–80% for the
bituminous coal and 20–30% for potassium ores; – comparatively high content of the useful component which is 60–80% for the
bituminous coal and 20–30% for potassium ores; p
;
– polydispersity of the impregnations of the useful component is from 0.5 to 10 mm;
l ti
l
l
d
it
f th
tit
t
k i f
1 4 t
2 3
/
3 – polydispersity of the impregnations of the useful component is from 0.5 to 10 mm;
– relatively low density of the constituent rocks is from 1.4 to 2.3 g/cm3; – polydispersity of the impregnations of the useful component is from 0.5 to 10 – relatively low density of the constituent rocks is from 1.4 to 2.3 g/cm3; – the presence of argillaceous material such as the layered silicates swelling in the
water. – the presence of argillaceous material such as the layered silicates swelling in the
water. 1
Introduction Many deposits of minerals are associated with the presence of layered silicates such as
the clay, mudstones, argillaceous slate - highly hydrophilic mineral varieties that have
the ability to swell in the water. This ability to self-disperse creates the great difficulties
both for the extraction and for the processing of such ores. Since the processing of most
minerals is associated with the use of wet processes, where in contact with water the
swelling layered silicates are dispersed to a particle size of less than 50 lm, but due to
their high hydrophylic property they form bonds that promote the formation of so-
called structured suspensions having increased viscosity. In this case, the structured
suspensions make it difficult to perform the separation operations as the classification,
the gravity and magnetic separation, the flotation and thickening and filtration opera-
tions (Arsentyev et al. 2014). In order to eliminate the influence of swelling silicates on the treatment processes it
is necessary to use the diluted suspensions with the solids content of 10–20% while the
usual solids content in processing suspensions is 35–50%. This causes the increase in
3–4 times the consumption of water during the processing of ores containing the
swelling sheet silicates (Zhdanovich et al. 2011). The most large-tonnage processed commercial mineral containing the swelling
silicates are the coal and the potassium salts. Arsentyev VA, Vaisberg LA, Ustinov ID (2014) Directions of creation of low-water technologies
and devices for the processing of finely divided mineral raw materials. Obogashenie Rud
(5):3–9 4
Conclusions An analysis of international practice has shown that fresh water is becoming the most
scarce resource for the mining and processing industry, which leads to a significant
increase in the cost of water supply and circulation systems, with their growth rates
being four to five times higher than the growth rates of the ore mining output. Studies of the problems of reducing water consumption in mineral processing
should be combined with energy consumption assessments due to the close relationship
between water and energy consumption values within a single processing system of
any plant. Most often, a technology that ensures water saving requires higher energy
consumption and vice versa. The presence of layered silicates with mobile crystal lattices in the mined ore
significantly complicates both its wet and dry processing. A promising solution, aimed at efficient use of resources in processing of ores
containing layered silicates, implies the inclusion of thermal and hydrothermal modi-
fication of such ores as the primary stages of the process chain. These modifications
would ensure a reduction or elimination of the negative impact of such materials
currently experienced in ore preparation, separation and dewatering and in the storage
of processing products. The solution requires additional energy inputs, but ensures significant savings in
other resources. Acknowledgements. Financial support was provided by the Russian Scientific Fund (project
18-17-00169). 3
Results and Discussion The thermal treatment of the high-ash coals (low-temperature pyrolysis reaction)
modefies the physicochemical properties of the minerals that make up the ash reducing
their ability to swell in water and increasing the magnetic susceptibility which makes it
possible to significantly intensify the processes of their magnetic-flotation processing to
produce the high-quality solid fuels. In the coal, under the influence of temperatures in
the process of the coal pyrolysis, the initial fracture structure is transformed into the
cracked-pore structure with the increase in the number and dimensions of the poriness,
vesicles and cracks. The sample of the coal subjected to medium-temperature pyrolysis
reaction compared to the initial coal has 20% lower apparent density and the specific
energy consumption of crushing is 20–30% lower. The specific ash content per unit of the heat of combustion indicates that the
combined dry processing of coal allows reducing the specific ash content by 1.3–1.6
times i.e. the formation of the ash and slag during the combustion is reduced. It has been established that the thermal treatment of the sylvinite ore; containing the
argllous-carbonate insoluble minerals in the range of 100–500 °C practically does not
affect the structure of salting minerals of the halite and the silvinite but it significantly A. Gerasimov et al. 68 modifies the structure of minerals entering into the insoluble fraction which has the
positive effect on the flotation of these ores and the process of sedimentation of the
slurry fractions in the flotation tails. For reducing the energy costs for the heat treatment of sylvinite ores it has been
proposed to use microwave treatment. The hydrothermal treatment of kaolin raw material in the autoclave can signifi-
cantly reduce the viscosity of kaolin suspensions and further efficient processing to
produce the concentrates for the production of alumina by using the wet screening
process for the suspended matters with the relatively high density. Arsentyev VA, Vaisberg LA, Ustinov ID, Gerasimov AM (2016) Prospects for reducing water
use in coal-processing. Mining J (5):97–100 Arsentyev VA, Gerasimov AM, Kotova EL (2017a) Thermochemical modification of sylvinite
ore using the microwave heating. Obogashenie Rud (6):3–8 Acknowledgements. Financial support was provided by the Russian Scientific Fund (project
18-17-00169). © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 70–74, 2019.
https://doi.org/10.1007/978-3-030-22974-0_16 1
Introduction Accessory minerals, in the classical sense, are minerals contained in rocks in small
quantities (less than 1%). It may be assumed that the term is also applicable to an
expanding range of mineral resources, including rare minerals of anthropogenic
deposits and mineral metallurgical wastes. Typical accessory minerals of sedimentary
rocks, metallurgical slags and calcines include sulphide minerals of iron, copper, and
zinc, often containing precious metals. This work covers certain chemical transformations of accessory sulphides con-
tained in the wastes of the metallurgical industry. Note that similar behaviors of sul-
phides may also be observed in the thermal processing of coal. Process mineralogy
(Bradsow 2014), recently expanded to include thermal processes (Rubin et al. 2014), is
the critical tool when studying the behavior of accessory sulfide minerals. Applied Mineralogy of Anthropogenic
Accessory Minerals A. Gerasimov1(&), E. Kotova2, and I. Ustinov1 1 REC «Mekhanobr-Tekhnika», St-Petersburg, Russia
gornyi@mtspb.com
2 Mining Museum, St-Petersburg Mining University, St-Petersburg, Russia Abstract. The term “accessory minerals” is applicable to an expanding range
of mineral resources, including rare minerals of anthropogenic deposits and
mineral metallurgical wastes. Typical accessory minerals of sedimentary rocks,
metallurgical slags and calcines include sulphide minerals of iron, copper, and
zinc. This work covers certain chemical transformations of accessory sulphides
contained in the wastes of the metallurgical industry. Note that similar behaviors
of sulphides may also be observed in the thermal processing of coal. It is shown
that, in terms of thermodynamics, thermal processing of products containing
precious metals enables application of various methods for generating artificial
covellite with silver and gold content through the use of accessory sulphide
minerals. Keywords: Accessory minerals Precious metals Thermal processing
Thermodynamics Applied Mineralogy of Anthropogenic
Accessory Minerals
A. Gerasimov1(&), E. Kotova2, and I. Ustinov1
1 REC «Mekhanobr-Tekhnika», St-Petersburg, Russia
gornyi@mtspb.com
Mining Museum, St-Petersburg Mining University, St-Petersburg, Russia Applied Mineralogy of Anthropogenic
Accessory Minerals
A. Gerasimov1(&), E. Kotova2, and I. Ustinov1
1 REC «Mekhanobr-Tekhnika», St-Petersburg, Russia
gornyi@mtspb.com
2 Mining Museum, St-Petersburg Mining University, St-Petersburg, Russia References Arsentyev VA, Vaisberg LA, Ustinov ID (2014) Directions of creation of low-water technologies
and devices for the processing of finely divided mineral raw materials. Obogashenie Rud
(5):3–9 Arsentyev VA, Gerasimov AM, Kotova EL (2017a) Thermochemical modification of sylvinite
ore using the microwave heating. Obogashenie Rud (6):3–8 Arsentyev VA, Vaisberg LA, Ustinov ID, Gerasimov AM (2016) Prospects for reducing water
use in coal-processing. Mining J (5):97–100 Properties and Processing of Ores Containing Layered Silicates 69 Arsentyev VA, Gerasimov AM, Mezenin AO (2017b) Study of technology of kaolin processing
using hydrothermal modification. Obogashenie Rud (2):3–9 Gerasimov AM, Dmitriev SV (2016) Combined technology of dry coal processing. Obogashenie
Rud (6):9–13 Lyskova MY (2016) Geoecology in the modern construction of enterprises for the extraction and
enrichment of potassium-magnesium salts, News of Tula State University. Geosciences
(4):39–49 Mozheiko FF, Potkin TN (2008) Physico-chemical basis of processing of high-arcilla off-balance
sylvinite ores. Vesti, National Academy of Sciences of Belarus, Series of Chemical Sciences,
no 4, pp 25–32 pp
Titkov SN et al (2013) The technology of dry crushing of potash ore to flotation size. In: IX
Congress of enrichers of the CIS countries. Collection of materials, vol 2. Digest MISA. M. 2013, pp 578–583 Zhdanovich IB et al (2011) Influence of heat treatment of saliferous arcilla on the structural-
geological properties of their dispersions. Vesti, National Academy of Sciences of Belarus,
Series of Chemical Sciences, no 3, pp 113–117 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches The traditional technology used for the processing of zinc concentrates obtained in
flotation concentration of polymetallic sulphide ores envisages oxidizing roasting of the
zinc concentrate, in which the main mineral, ZnS sphalerite, is converted into zinc Applied Mineralogy of Anthropogenic Accessory Minerals 71 sulphate ZnSO4. The inevitable impurities of pyrite FeS2 and chalcopyrite CuFeS2,
present in the zinc concentrates, are also oxidized into the corresponding simple sul-
fates and oxides at temperatures above 770 °K (Naboichenko et al. 1997). At present,
fluidized bed furnaces are predominantly used for oxidizing roasting, with the oxida-
tion process occurring in a hot gas flow in a suspended state. Despite the obvious benefits, such as high yield and roasting rates, fluidized bed
furnaces also have certain disadvantages, including partial underburning of sphalerite
due to the rapid formation of a poorly gas-permeable zinc sulfate film on its surface. This inevitably leads to underburning of fine inclusions in sphalerite grains of other
sulfide minerals, primarily chalcopyrite. Chalcopyrite is the most common impurity in
sphalerite, which is due to the high similarity of the structures of their crystal lattices. It
is known that chalcopyrite is most frequently found in the sphalerite structure in the
form of very fine grains, down to emulsion shots (Betekhtin 2008). The behavior of
chalcopyrite inclusions in sphalerite is significant for subsequent stages of processing
of the calcined concentrate. After the oxidizing roasting, the resulting zinc calcine is subjected to staged sulfuric
acid and aqueous leaching of zinc sulfate and oxide-sulfate with the recovery of a zinc
sulfate solution to be subsequently used for zinc metal precipitation by electrolysis. Acid and neutral zinc sulfate leaching is performed at temperatures of up to 80–100 °C,
additionally enabling chemical transformations of insoluble calcine components on the
grain surfaces. The insoluble cake obtained after leaching includes a certain amount of
underburned sulphide minerals containing precious metals, primarily silver. It was
previously demonstrated (Otrozhdennova et al. 1997; Geikhman et al. 2003) that
sulphide minerals remaining in the cake may be flotation concentrated, yielding a froth
product containing up to 18% of sulfide sulfur and up to several kilograms of silver per
ton. We have studied the composition of concentrates obtained by flotation concen-
tration of the cake generated after zinc sulphate leaching at three metallurgical enter-
prises in different countries. 2
Methods and Approaches The studies were performed using a Camscan scanning
microscope, an Analysette laser microanalyzer, a Geigerflex X-ray phase spectrometer,
optical microscopy tools, etc. It was found that there was no fundamental difference in the material composition
of such concentrates at different metallurgical plants. This is explained by the similarity
of the ore bases and of the metallurgical treatment structures used at the mining
enterprises analyzed. Sphalerite (70–80%) is the main mineral found in the composition
of the concentrates, followed by such spinel-structure minerals as MeFe2O4, covellite,
oxide compounds of zinc and iron (several percent), and traces of chalcosite. In terms of their elementary chemical composition, the concentrates were repre-
sented by zinc 45–48%, total sulfur 20–22%, sulfur sulfide 17–19%, iron 11–14%,
silica 2.5–4%, and copper 3–4%. The silver content was 0.2–0.4% (2–4 kg/t); the gold
content was 10–15 g/t. 72 A. Gerasimov et al. 3
Results and Discussion The most important finding is the presence of copper sulphide covellite CuS in the
flotation concentrate, in which the bulk of silver is concentrated. The availability of
silver-containing films of copper sulfides (without their identification) on the surfaces
of sphalerite contained in the zinc cake flotation concentrate has been indicated pre-
viously (Otrozhdennova et al. 1997). We have established that covellite forms distinct
substances on the sphalerite surface. No native silver was found in the samples. Gold is
present in the form of fine grains containing silver and copper. The gold sample is 700–
930. Copper-containing metals in the source zinc concentrate entering the roasting
process are represented almost exclusively by chalcopyrite, including its aggregates
with sphalerite. Two main methods of covellite formation in the metallurgical processing of zinc
may be considered in this regard. The first is the interaction of dissolved copper sulfate
with the sphalerite surface in sulfuric acid leaching of zinc calcines, with the formation
of covellite films in the course of the resulting exchange reaction and with the transition
of zinc ions into the solution. The second possible source of covellite is the transfor-
mation of fine chalcopyrite inclusions contained in sphalerite grains with the migration
of the resulting covellite to the sphalerite surface due to the strong differences in the
structures of their crystal lattices. None of these methods has been subjected to special
experimental verification. Given that even isomorphous substitution of copper with
silver can occur in chalcopyrite, despite the difference in the electron spiral radii of
silver of 1.26 Å and copper of 0.96 Å, it may be assumed that silver-containing
covellite may be generated using these methods, especially considering the effects
produced on such processes by associated minerals. Let us consider the thermodynamic feasibility of the two hypothetical processes of
formation of man-made covellite. In the below calculations of the free energy of the
processes and their enthalpy and entropy increments, known reference data are used for
reactions (I) and (II). These calculations show that reaction (I) can occur spontaneously,
which is facilitated by the entropy effect, and reaction (II) requires external energy
inputs, which does not contradict the conditions of respective production processes
(Tables 1 and 2). Table 1. Acknowledgements. The work was carried out with the support of the Russian Science
Foundation (project No. 17-17-30015). Table 2. Thermodynamic characteristics Table 2. Thermodynamic characteristics
Substance DH0, kJ/mol S0
298, kJ/mol K CP, kJ/mol K
CuFeS2
−194.93
124.9
95.67
CuS
53.10
66.5
47.8
FeS
−100.00
60.3
50.5
Peaкция
41.8
1.9
2.63 ZnS + CuSO4 ! CuS + ZnSO4
ð1Þ ð1Þ DH0
798 ¼ DH0
298 þ
Z798
298
DcpdT ¼ 58:5 103 þ 2:1 ð798 298Þ ¼ 57:45 kJ/mol
DS0
798 ¼ DS0
298 þ
Z798
298
Dcp
T ¼ 10:1 þ 2:1 ln 798
298 ¼ 12:17 J/molK DH0
798 ¼ DH0
298 þ
Z798
298
DcpdT ¼ 58:5 103 þ 2:1 ð798 298Þ ¼ 57:45 kJ/mol
798 DS0
798 ¼ DS0
298 þ
Z798
298
Dcp
T ¼ 10:1 þ 2:1 ln 798
298 ¼ 12:17 J/molK 298
DG0
798 ¼ DH0
798 798 DS0
798 ¼ 57450 798 12:17 ¼ 67:2 kJ/mol CuFeS2 ! CuS + FeS2
ð2Þ ð2Þ CuFeS2 ! CuS + FeS2 DH0
798 ¼ DH0
298 þ
Z798
298
DcpdT ¼ 43; 15 kJ/mol
DS0
798 ¼ DS0
298 þ
Z798
298
Dcp
T
¼ 4; 4 J/molK
DG0
798 ¼ DH0
798 798 DS0
798 ¼ 39; 56 kJ/mol DS0
798 ¼ DS0
298 þ
Z798
298
Dcp
T
¼ 4; 4 J/molK DG0
798 ¼ DH0
798 798 DS0
798 ¼ 39; 56 kJ/mol Applied Mineralogy of Anthropogenic Accessory Minerals Applied Mineralogy of Anthropogenic Accessory Minerals 73 Table 2. Thermodynamic characteristics 3
Results and Discussion Thermodynamic characteristics
Substance DH0, kJ/mol S0
298, J/mol K CP, kJ/mol K
ZnS
−206
57.7
46.0
CuSO4
−771.4
109.2
98.9
CuS
53.1
66.5
47.8
ZnSO4
−982.8
110.5
99.2
Peaкция
−58.5
10.1
2.1 Table 1. Thermodynamic characteristics Applied Mineralogy of Anthropogenic Accessory Minerals 4
Conclusions In terms of thermodynamics, combined thermal processing of products containing
precious metals enables application of various methods for generating artificial cov-
ellite with the participation of accessory sulphide minerals. Respective statements may
be useful for both metallurgical processes and thermochemical processing of hard
coals, which always contain impurities of sulphide minerals. Acknowledgements. The work was carried out with the support of the Russian Science
Foundation (project No. 17-17-30015). 74 A. Gerasimov et al. Naboichenko SS (1997) Processes and devices of non-ferrous metallurgy. USMU, Ekaterinburg
Rubin S, Aksenov A, Senchenko A, Lagutina S (2014) Argentojaroside synthesis and research.
In: Proceedings of the XXVII IMPC, C.19, pp 209–213 1
Introduction The most important geochemical feature of iron is the presence of several oxidation
degrees. By crystal-chemical properties of the Fe2+ ion is close to the Mg2+ and Ca2+
ions and to the other main elements. Due to crystal-chemical similarity Fe replaces Mg
and partially Ca in many silicates. In quartzites, the main ore mineral is magnetite, the
crystal-chemical structure characteristics of which determine the technological prop-
erties of quartzites, according to which quartzites are characterized by high variability
of composition and properties, and the results of enrichment depend on their material
composition. It is worth noting the diversity of mineral composition, texture and
structural features of ores and, as a consequence, a wide range of physical, mechanical
and technological properties (Gsogyan 2010). References Betekhtin AG (2008) Course of Mineralogy. Publishing House University, Moscow (2008)
Bradsow D (2014) The role of ‘Process mineralogy’ in improving the process performance of
complex sulfide ores. In: Proceedings XXVII IMPC, C.14, pp 1–23
Geikhman VV, Kazanbaev LA, Kozlov PA (2003) Industrial tests of flotation of zinc cakes. Non-
ferrous Met (1):29–32
Otrozhdennova LA, Maksimov II, Khodov NV (1997) Combined technology of silver recovery
from zinc cakes. Min J (4):39–40
Naboichenko SS (1997) Processes and devices of non-ferrous metallurgy. USMU, Ekaterinburg
Rubin S, Aksenov A, Senchenko A, Lagutina S (2014) Argentojaroside synthesis and research. In: Proceedings of the XXVII IMPC, C.19, pp 209–213 Betekhtin AG (2008) Course of Mineralogy. Publishing House University, Moscow (2008) gy
g
y
Bradsow D (2014) The role of ‘Process mineralogy’ in improving the process performance of
complex sulfide ores. In: Proceedings XXVII IMPC, C.14, pp 1–23 Bradsow D (2014) The role of ‘Process mineralogy’ in improving the pro
complex sulfide ores. In: Proceedings XXVII IMPC, C.14, pp 1–23 Geikhman VV, Kazanbaev LA, Kozlov PA (2003) Industrial tests of flotation of zinc cakes. Non-
ferrous Met (1):29–32 Otrozhdennova LA, Maksimov II, Khodov NV (1997) Combined technology of silver recovery
from zinc cakes. Min J (4):39–40 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Crystal-Chemical and Technological Features
of the KMA Natural Magnetites T. Gzogyan(&) and S. Gzogyan Belgorod National Research University, Belgorod, Russia
mehanobr1@yandex.ru Belgorod National Research University, Belgorod, Russia
mehanobr1@yandex.ru Abstract. The results of researches of crystal-chemical features of magnetites
obtained from the KMA ferruginous quartzites deposits are presented. It is
shown that all magnetites are represented by cation-scarce mineral differences. The correlation dependences between the technological indexes (yield, extrac-
tion) and the magnetite concentration in quartzites are established. Keywords: Crystal-chemical features Ferruginous quartzites Magnetite
Maghemite Mossbauer spectra Magnetic properties Defectiveness © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 75–79, 2019.
https://doi.org/10.1007/978-3-030-22974-0_17 2
Methods and Approaches In this work the crystal-chemical features of the main ore minerals of unoxidized and
oxidized quartzite KMA deposits are investigated using modern research methods
(nuclear gamma-resonance spectroscopy (Mossbauer effect), X-ray analysis and X-ray
fluorescence). Measurements of the dependence of magnetization on the applied
magnetic field and saturation magnetization by the vibration method are carried out. Magnetite is a typomorphic mineral composition and properties of which depends
on the conditions of formation and has got a structural spinel. This is the ohFd3 m
space group, where cations occupy 16 octahedral (B) and 8 tetrahedral (A) positions. Therefore, for ideal crystals of Fe3 þ
1
[Fe3 þ
1
F2 þ
1
O4 the reversed spinel, the ratio of the
intensities of the leftmost lines of the mossbauer absorption spectra, taking into account T. Gzogyan and S. Gzogyan 76 the difference in the probabilities of the effect for the octahedral and tetrahedral sub-
lattices, should be slightly less than two. 3
Results and Discussion The appearance of a cationic
vacancy in position B of the spinel structure does not lead to experimentally observed
broadening but only to a redistribution of the intensities of the lines corresponding to
Fe3+ and [Fe3+, Fe2+] in positions A and B due to increasing in the contribution of Fe3+
which does not participate in electronic exchange Fe2+ with position B due to
vacancies. But in both cases the integral intensity remains constant. However, the
presence of a cationic vacancy in the octahedral position in the spinel structure sig-
nificantly affects the technological properties. The combination of experimental data
obtained allows us to write the crystal-chemical formula of magnetites in the form
Fe3 þ
1
Fe3 þ
5=32=3XFe2 þ
xh1=31=3X
h
i
O4, where □– is a cation vacancy (in the structure of
magnetite as isomorphic substituting ions can be Mg+2, Mn+2, Ni+2, Ti+2, etc.), and x –
is non-stoichiometric (the defectiveness is the Fe2+ deficiency widely varying in area of
deposits). At x = 0 we have a six-peak absorption spectrum corresponding to
maghemite. The condition of electro-neutrality is observed due to the formation of
defects that is, electron holes. The relationship between Fe3+ and [Fe3+, Fe2+] deter- appearance of isomorphically replacing ions in the octahedral position of the magnetite
spinel structure leads to a partial removal of electron exchange in position B between
Fe2+ and Fe3+ and, as a result, to a change in the ratio of the intensities of the lines
corresponding to iron ions in positions A and B with simultaneous broadening of not
only the extreme right groups of lines, but also the entire set of lines of the total
resonance absorption spectrum (broadening is especially pronounced with an increase
in the concentration of isomorphically substituting ions). The appearance of a cationic
vacancy in position B of the spinel structure does not lead to experimentally observed
broadening but only to a redistribution of the intensities of the lines corresponding to
Fe3+ and [Fe3+, Fe2+] in positions A and B due to increasing in the contribution of Fe3+
which does not participate in electronic exchange Fe2+ with position B due to
vacancies. But in both cases the integral intensity remains constant. However, the
presence of a cationic vacancy in the octahedral position in the spinel structure sig-
nificantly affects the technological properties. 3
Results and Discussion The characteristic lines of the obtained Mössbauer absorption spectra of quartzites
represent the superposition of a series of six-peak spectra. The ratio of the intensities
corresponding to half of the ions in the A and B positions is practically not satisfied,
despite the fact that the selected sample is closest to the monomineral difference. In
addition, there is also a slight broadening of the leftmost and rightmost lines of the total
absorption. Mossbauer spectra indicate that the intensity ratio 1:2 is not satisfied. Violation of
this ratio may be due either to an isomorphic substitution of iron ions in the B-position of Fe3 þ
1
Fe3 þ
5=32=3XF2 þ
XYMe2 þ
yh1=31=3X
h
i
O4 or to the phenomenon of non-stoichiometry
and
the
presence
of
Fe1
2+
vacancies
in
the
same
position
Fe3 þ
1
Fe3 þ
5=32=3XF2 þ
XYMe2 þ
yh1=31=3X
h
i
O4. Obviously, at x = 1, this is the stoichiometric
magnetite Fe3 þ
1
[Fe3 þ
1
F2 þ
1
O4, and at x = 0 – c – Fe2O3 Fe3 þ
1
Fe3 þ
5=3h2=3X
h
i
O4
. The of Fe3 þ
1
Fe3 þ
5=32=3XF2 þ
XYMe2 þ
yh1=31=3X
h
i
O4 or to the phenomenon of non-stoichiometry
and
the
presence
of
Fe1
2+
vacancies
in
the
same
position
Fe3 þ
1
Fe3 þ
5=32=3XF2 þ
XYMe2 þ
yh1=31=3X
h
i
O4. Obviously, at x = 1, this is the stoichiometric
magnetite Fe3 þ
1
[Fe3 þ
1
F2 þ
1
O4, and at x = 0 – c – Fe2O3 Fe3 þ
1
Fe3 þ
5=3h2=3X
h
i
O4
. The appearance of isomorphically replacing ions in the octahedral position of the magnetite
spinel structure leads to a partial removal of electron exchange in position B between
Fe2+ and Fe3+ and, as a result, to a change in the ratio of the intensities of the lines
corresponding to iron ions in positions A and B with simultaneous broadening of not
only the extreme right groups of lines, but also the entire set of lines of the total
resonance absorption spectrum (broadening is especially pronounced with an increase
in the concentration of isomorphically substituting ions). 3
Results and Discussion The combination of experimental data
obtained allows us to write the crystal-chemical formula of magnetites in the form
Fe3 þ
1
Fe3 þ
5=32=3XFe2 þ
xh1=31=3X
h
i
O4, where □– is a cation vacancy (in the structure of
magnetite as isomorphic substituting ions can be Mg+2, Mn+2, Ni+2, Ti+2, etc.), and x –
is non-stoichiometric (the defectiveness is the Fe2+ deficiency widely varying in area of
deposits). At x = 0 we have a six-peak absorption spectrum corresponding to
maghemite. The condition of electro-neutrality is observed due to the formation of
defects that is, electron holes. The relationship between Fe3+ and [Fe3+, Fe2+] deter-
mines the behavior of such magnetites in magnetic fields. Fe3 þ
1
Fe3 þ
5=32=3XFe2 þ
xh1=31=3X
h
i
O4, where □– is a cation vacancy (in the structure of
magnetite as isomorphic substituting ions can be Mg+2, Mn+2, Ni+2, Ti+2, etc.), and x –
is non-stoichiometric (the defectiveness is the Fe2+ deficiency widely varying in area of
deposits). At x = 0 we have a six-peak absorption spectrum corresponding to
maghemite. The condition of electro-neutrality is observed due to the formation of
defects that is, electron holes. The relationship between Fe3+ and [Fe3+, Fe2+] deter-
mines the behavior of such magnetites in magnetic fields. In addition, martite lines are present on the Mossbauer spectra of magnetites and
this indicates that the crystal lattice of magnetites is coherently connected with the
lattice
of
the
“parent”
magnetite. The
presence
of
a
coherently
coupled Crystal-Chemical and Technological Features of the KMA Natural Magnetites 77 Fe0
1 Fe3 þ
5=3h1=3
h
i
O4 structure significantly affects the degree of its extraction during the
enrichment. Traces of hematite and various non-metallic particles are present with
magnetite. Thus, magnetite is a very sensitive indicator of the conditions of deposits formation
and its “life”, started in nature, is manifested and sometimes significantly in the kinetics
of technology, because rapid processes intensively destroy its information structure and
slowly transfer it to the products resulting from processing (Gsogyan 2010). Features of heterogeneity of the composition and properties of magnetite affect the
variability of its oxidation degree in the course of crushing process and, as a result, the
technological indicators of enrichment (Gsogyan 2010). Quantitative XRF of quartzites
revealed the presence in them along with magnetite, hematite and maghemite; more-
over, the presence of maghemite is the characteristic of all mineral types of quartzite. 3
Results and Discussion XRF showed that the highest content of maghemite (*11%) is the characteristic of
magnetite quartzite, somewhat lower (*8%) is observed in semi-oxidized and the
lowest (*4.5%) in oxidized. Morphological series observed in nature in ferruginous quartzites: Fe4O4 (wus-
tite) ! Fe2+Fe2
3+O4
(magnetite) ! cFe2O3
(maghemite) ! aFe2O3
(he-
matite) ! Fe2O3H2O
(goethite) ! cFeO(OH)
(lepidocrocite) ! Fe2O3xH2O
(limonite /martite) emphasizes the complex relationships between their structures. With
the transition from one structural type to another, some features of the original type are
inherited. The edge of the unit cell, depending on the composition of the oxidation of
wustite to magnetite and maghemite consistently decreases with the number of Fe2+
cations. Linear dependence of the lattice parameter on the composition emphasizes the
similarity between the structures of these compounds. Reducing edges of the unit cell
of magnetite in the transition to maghemite caused by the replacement of Fe2+ with an
ionic radius of 0,80 Å on Fe3+ with an ionic radius of 0,67 Å with simultaneous
removal of one third of the Fe2+ ions from the structure of magnetite. The magnetites
associated with the incomplete processes of martitization are highly heterogeneous,
which is associated with the manifestation of morphotropic transformations in the
series indicated above, the content of Fe in which falls from 72.4% (magnetite) to
62.0% (limonite). The average edge size of the unit cell varies from 8.371 Å (Lebedinsky) to 8.395 Å
(Mikhailovsky) and this is somewhat less than that of a magnetite of stoichiometric
composition (8.396 Å). The evaluation of magnetite deformations allows us to con-
clude that a change in the main interplanar distances of magnetite samples is more
characteristic of Lebedinsky and Prioskolsky. Microprobe analysis revealed iron deficiency in magnetite grains of various gen-
erations, the content of which varies from 66.89% (Mikhailovsky) to 72.3%
(Stoilensky). Thermomagnetic analysis in almost all magnetites at T = 340–350 °C shows a
weak exothermic effect corresponding to maghemite and its presence is confirmed by
electron diffraction patterns, which indicate the absence of periodicity in the arrange-
ment of some reflexes and the presence of defects and dislocations in the magnetite
lattice (Fig. 1). 78 T. Gzogyan and S. Gzogyan Elem Wt % Mol %
SiO2
2.26 5.79
Fe2O3 97.74 94.21
Total 100.00 100.00
Fig. 1. The electron phase, taken from the magnetite particles at different angles of orientation
of the device
78
T. Gzogyan and S. Gzogyan Fig. 3
Results and Discussion The Crystal-Chemical and Technological Features of the KMA Natural Magnetites 79 obtained dependence of the specific saturation magnetization has an inflection point
which apparently is associated with the influence of a sufficiently large weight fraction
of Fe3 þ
1
Fe3 þ
5=3 Fe2 þ
1=3
h
i
O4 The unevenness of magnetic properties is due to the change in
the ratio of Fe3+ and Fe2+. Specific magnetic susceptibility indicates a different pet-
rographic composition of ores and the size of magnetite impregnation and their sig-
nificant heterogeneity which is a consequence of their genesis and is expressed in the
difference in their technological properties. The saturation magnetization is a multi-
parameter function for ferruginous quartzites. In this regard, the dependences of technological indicators of enrichment were
obtained as a function of the reduced value (rs/rs
id), where rs are the obtained values
of the specific magnetization of this sample; rs
id is the specific magnetization of the
isolated magnetite fraction (Fetot = 72.2%, SiO2 less than 0.2%). 3
Results and Discussion 1. The electron phase, taken from the magnetite particles at different angles of orientation
of the device Maghemite is marked from 11% (Mikhailovsky and Prioskolsky) to 6% (Kor-
obkovsky). The data of high-temperature magnetometry confirm the lack of Fe+2 in the
KMA magnetites and their difference from stoichiometric: the Curie point for mag-
netite averaged 571.8 Å (Stoilensky) to 578.3 Å (Prioskolsky). The change in the ratio
of Fe3+ and Fe2+ causes the inextensive magnetic properties which are one of the most
important factors affecting the enrichment. These studies suggest that, due to the lack of
a sufficient amount of isomorphic impurities, we are dealing with alternative behavior
in the two-phase system of Fe2O3 Fe3O4, that is, the formation of metastable tran-
sitional phases with a structure closer to the matrix than the equilibrium phase. This is
confirmed by the results of determining the concentrations of isomorphically substi-
tuting impurities of Mg2+, Mn2+, Ti2+ and Ni2+, which did not exceed 0.2% NiO;
0.25% MnO2 and 0.4% MgO. All Mossbauer absorption spectra contain maghemite lines. Consequently, the
formation of solid solutions of FeO:Fe2O3 coherently coupled with c-Fe2O3 and semi-
coherently with Fe2O3 is strictly observed in deposits. This led to the widespread
development of quartzites heterogeneity in their composition and properties. The
coherent phases formed in this way can be clusters of dissolved atoms in the lattice of
the parent material that is often observed. The presence of “non-stoichiometric” cation-
deficient differences of magnetites led to the need to find the value that determines
their
defectiveness. The
crystal-chemical
formula
of
such
magnetites
is:
Fe3 þ
1
Fe3 þ
5=32=3XFe2 þ
x
Fe2 þ
1=31=3X
h
i
O4 where x is defectiveness (Fe2+ deficiency in the
octahedral
position
of
the
spinel
structure)
(for
x = 1
is
ideal
magnetite
Fe3 þ Fe3 þ
1
Fe2 þ
O4 (content of 72.4%), and at x ! 0 a complete transition up to c-
Fe2O3 (crystallographic symmetry) Fe3 þ
1
Fe3 þ
5=3 Fe2 þ
1=3
h
i
O4 when a part of iron ions is
recharged to the state of Fe3+ and the iron content drops to 69.9%). The dependence of the magnetic properties on the composition and crystal-
chemical features of magnetites is performed by the vibration method in a uniform
magnetic field H up to 71.62 kA/m in the temperature range from 20 to 800 °C. 4
Conclusions – magnetites from ferruginous quartzite deposits of the KMA are represented by
cation-deficient mineral differences with a wide variation of the magnitude of
defects (x) in the area of occurrence; – the presence of coherently bound c-Fe2O3 and semi-coherently bound a-Fe2O3 is
determined depending on the degree of oxidation which undergoes spinodal and
intracrystalline decomposition up to hematite; – technological indicators of enrichment depends on the crystal-chemical and mag-
netic properties of quartzites. These allow to find optimal conditions for concen-
trates obtaining, i.e. implement the “geological” management of the production. – technological indicators of enrichment depends on the crystal-chemical and mag-
netic properties of quartzites. These allow to find optimal conditions for concen-
trates obtaining, i.e. implement the “geological” management of the production. Gzogyan TN (2010) To the question of the heterogeneity of the KMA magnetite deposits. Min
Inf-Anal Bull (5):256–259 Gzogyan TN (2010) To the question of the heterogeneity of the KMA magnetite deposits. Min
Inf-Anal Bull (5):256–259 Gzogyan TN (2010) To the question of the heterogeneity of the KMA magnetite deposits. Min
Inf-Anal Bull (5):256–259 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction Actual problems of processing iron ores and quartzites remain increasing in the met-
allurgical value of the concentrate, reducing metal losses and costs in the production of
commercial products. In increasing the efficiency of processing ferruginous quartzites a
special role belongs to the scientific direction – the technological mineralogy. The
transition from the descriptive methodology to the methodology of genetic analysis
allowed giving a scientific interpretation of many problems associated with the tech-
nology of processing ferruginous quartzites. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 80–83, 2019.
https://doi.org/10.1007/978-3-030-22974-0_18 Practical Application of Technological
Mineralogy on the Example of Studying
of Sulphidization in the KMA
Ferruginous Quartzites S. Gzogyan(&) and T. Gzogyan
Belgorod National Research University, Belgorod, Russia
mehanobr1@yandex.ru S. Gzogyan(&) and T. Gzogyan Belgorod National Research University, Belgorod, Russia
mehanobr1@yandex.ru Abstract. The results of mineralogical-petrographic researches of relationship
of iron sulphidic minerals with a magnetite and their influence on the techno-
logical properties of the KMA ferruginous quartzites are presented. A classifi-
cation of ferruginous quartzites by sulphidic factor on the basis of textural and
structural features of the divided mineral components is developed. Keywords: Ferruginous quartzites Pyrite Pyrrhotine Magnetite
Sulphidic factor Keywords: Ferruginous quartzites Pyrite Pyrrhotine Magnetite
Sulphidic factor Practical Application of Technological Mineralogy 81 2
Methods and Approaches The object of a research is more than 900 group geological and technological samples
of the KMA ferruginous quartzites obtained during the operational exploration of
deposits, which reflect the most representative texture-structural and mineralogical
features of the KMA quartzite deposits. The study of the composition and properties of the samples taken was carried out
using
optical
microscopy,
NMR-spectroscopy,
high-temperature
magnetometry,
microprobe analysis and X-ray diffractometry. The physical properties are used to determine the microhardness of minerals and
magnetic properties. The main attention is paid to relationship of iron oxides and
sulphides and their distribution in the finished product. Researches were carried out on
samples of polished sections and briquettes. Practical Application of Technological Mineralogy 3
Results and Discussion The object of a research is more than 900 group geological and technological samples
of the KMA ferruginous quartzites obtained during the operational exploration of
deposits, which reflect the most representative texture-structural and mineralogical
features of the KMA quartzite deposits. The study of the composition and properties of the samples taken was carried out
using
optical
microscopy,
NMR-spectroscopy,
high-temperature
magnetometry,
microprobe analysis and X-ray diffractometry. The physical properties are used to determine the microhardness of minerals and
magnetic properties. The main attention is paid to relationship of iron oxides and
sulphides and their distribution in the finished product. Researches were carried out on
samples of polished sections and briquettes. The second category (medium enriched) includes ferruginous quartzites in which
the structure of relationship between ore minerals and sulphidic minerals is more
complex, that contributes to the transition to magnetite concentrate of both free grains
of pyrrhotite and aggregates of magnetite with pyrite (Fig. 1b). From ferruginous
quartzites of the second category (sulfur content 0.18), in the laboratory concentrate
0.09% remains (in the industrial one – 0.085%). Fig. 1. Pyrite in magnetite aggregates (a) and internal and external intergrowths of pyrite with
magnetite, poikilitic inclusions of pyrrhotite in magnetite (b), zoom 12.5 8 10. Fig. 1. Pyrite in magnetite aggregates (a) and internal and external intergrowths of pyrite with
magnetite, poikilitic inclusions of pyrrhotite in magnetite (b), zoom 12.5 8 10. The third category of quartzites (hardly enriched) is represented by ferruginous
quartzites with close structural intergrowths of sulphidic minerals with magnetite,
poikilitic inclusions of pyrite in magnetite and pyrrhotite and vice versa, and that leads
to the transition and concentration of sulphides in the process of wet magnetic sepa-
ration into magnetite concentrate (Fig. 2). With a total sulfur content in the feedstock of 0.42–0.47 in the laboratory magnetite
concentrate, its size reaches 0.42–0.78% (in the industrial one – 0.40–0.74%). Thus, optical and mineralogical researches have shown that clogging of magnetite
concentrate with sulfur occurs not only because of the ferromagnetic properties of the
monoclinic pyrrhotite variety, but also due to the inclusion of pyrite grains in the
magnetite and pyrrhotite grains and vice versa (Table 1). S. Gzogyan and T. Gzogyan 82 Fig. 2. Substitution structures of magnetite and pyrrhotite by pyrite, zoom 25 8 10 (a),
12.5 12.5 10 (b). Fig. 2. Practical Application of Technological Mineralogy 83 3
Results and Discussion Substitution structures of magnetite and pyrrhotite by pyrite, zoom 25 8 10 (a),
12.5 12.5 10 (b). Optical and mineralogical researches of finished products have established that
sulphidic minerals in concentrate are as in the form of intergrowths and inclusions in
magnetite grains as in the form of separate grains (Fig. 3). Table 1. Grain composition and distribution of chemical components by grain size in the
magnetite concentrate
Grain size, mm
Yield of grain size,% Content, %
Fe
S
Aggregate Magnetite Pyrite
Pyrrhotine
+0.071
1.20
29.4
27.53
0.046
0.040
−0.071 + 0.045
4.20
55.29
33.37
0.024
0.018
−0.045 + 0.032
27.33
66.92
64.88
0.028
0.019
−0.032 + 0.020
45.03
71.05
66.55
0.023
0.016
−0.020
22.24
70.47
62.77
0.021
0.014
Total
100.00
68.57
63.39
0.0242 0.0168 able 1. Grain composition and distribution of chemical components by grain size in th
agnetite concentrate Fig. 3. Magnetite concentrate: aggregates of iron sulphides with magnetite and their separate
grains (a, b) stand out against the background of free magnetite grains, poikilite inclusions in
magnetite (c), polished briquette, zoom 132. Fig. 3. Magnetite concentrate: aggregates of iron sulphides with magnetite and their separate
grains (a, b) stand out against the background of free magnetite grains, poikilite inclusions in
magnetite (c), polished briquette, zoom 132. Practical Application of Technological Mineralogy 4
Conclusions Thus, applying the methods of technological mineralogy and based on the studied
nature of the variability of structural transformations, the classification of quartzites
was developed according to enrichment by the sulphidic factor. The next important achievement of technological mineralogy in the context of a
particular sulphidic problem is the clearly shown migration of non-magnetic iron
sulphides into magnetite concentrate using magnetic separation processes. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Ore Mineralogy of Kirazliyayla (Yenişehir-
Bursa-Turkey) Mesothermal Zn-Pb-(–Cu)
Deposit: Preliminary Results F. Javid and E. Çiftçi(&) Department of Geological Engineering, Faculty of Mines, ITU, 33469 Maslak,
Istanbul, Turkey
eciftci@itu.edu.tr Abstract. The Kirazlıyayla deposit is one of Pb-Zn (±Cu) deposits associated
with andesitic volcanism cutting through the metamophics of Karakaya com-
plex. It is a structurally controlled, vein-stockwork style, mesothermal ore
deposit. Replacement, brecciation, vein/veinlets, carries, sea-island, and dis-
semination textures were identified. Pyrite, sphalerite, chalcopyrite, galena,
tennantite, and covellite constitute the ore mineral paragenesis. Quartz, calcite
and dolomite with kaolinite account for the gangue minerals. Supergene stage is
insignificant. The Kirazlıyayla mineralization is a Zn-Pb (±Cu) mineralization
hosted by tectonically controlled andesitic volcanism within the Karakaya
metamorphics and has common features with the other occurrences in the
western Anatolia. Keywords: Kirazlıyayla Mesothermal Lead Zinc Karakaya complex
Structurally controlled Keywords: Kirazlıyayla Mesothermal Lead Zinc Karakaya complex
Structurally controlled Ore Mineralogy of Kirazliyayla (Yenisehir-Bursa-Turkey) Although Turkey produces zinc and lead concentrates, due to the lack of smelters,
about 1.5 billion dollars each year paid for the import of those metals. In the recent
years, the government is in an attempt to revive the sector through various incentives. Although Turkey produces zinc and lead concentrates, due to the lack of smelters,
about 1.5 billion dollars each year paid for the import of those metals. In the recent
years, the government is in an attempt to revive the sector through various incentives. The western Anatolia is host to great number of mineral deposits. As for the lead-zinc
occurrences,itcanbesummarizedin3typesintermsoftheirgenesisintheregion:(I)distal
skarn occurrences associated with the carbonates of the metamorphic basement;
(II) structurally controlled veins within the basement metamorphics and the overlying
volcanics or along their contacts; and (III) replacements along the mafic dykes. The study area is located in one of the tectonic elements of Turkey – the Sakarya
terrane, which is an elongate crustal ribbon extending from the Aegean in the west to
the Eastern Pontides in the east. It is consisted of sandstones of Lower Jurassic age,
which sits on a fairly complex metamorphic basement that contains a high-grade
Variscan basement metamorphics of Carboniferous age (Topuz et al. 2004, 2007; Okay
et al. 2006), Paleozoic granitoids (Delaloye and Bingöl 2000; Okay et al. 2002, 2006;
Topuz et al. 2007), and a low grade metamorphic complex - the Lower Karakaya
Complex constituted by Permo-Triassic metabasite with lesser amounts of marble and
phyllite. The Lower Karakaya Complex represents the Permo-Triassic subduction-
accretion complex of the Paleo-Tethys as indicated by the presence Late Triassic
blueschists and eclogites (Okay and Monié 1997; Okay et al. 2002), accreted to the
margin of Laurussia during the Late Permian to Triassic. The complex is overlain by a
thick series of strongly deformed clastic and volcanic rocks with exotic blocks of
Carboniferous and Permian limestone and radiolarian chert. This complex basement
was overlain unconformably in the Early Jurassic by a sedimentary and volcanic
succession. The Early Jurassic is represented by fluvial to shallow marine sandstone,
shale and conglomerate in the western part of the Sakarya Zone. The metamorphic
basement is cut by Eocene volcanisms. The Kirazliyayla Zn-Pb ore deposit is spatially and temporally related with Eocene
intermediate extrusive rocks - andesite and trachyandesite with NE-SW extension and
covered by clastic and carbonate rocks. Ore Mineralogy of Kirazliyayla (Yenisehir-Bursa-Turkey) The main purpose of the investigation is to
determine the genesis of Kirazliyayla Zn-Pb ore deposit and its place in the metallo-
genic evolution of the region to contribute the understand of the metallogeny of Tur-
key. In that, geochemical characteristics, mineralogy of both host rocks and ore
minerals, its tectonic setting are the main issues are covered. 1
Introduction Turkey hosts noteworthy variety of mineral deposits owing to its geological evolution
within the Alpine-Himalayan orogenic belt and its complex tectonic setting. Mineral
deposits of Turkey are better understood through the understanding tectono-magmatic
evolution of Turkey. Within this framework, there are a number of various ore deposits
disseminated throughout the country including the Kuroko-type VMS deposits (strictly
in the Eastern Pontides tectonic belt), Cyprus-type VMS deposits (along the Bitlis-
Zagros suture zone in SE Anatolia and Küre-Kargı trend in the Central Pontides),
Besshi-type VMS deposits (in the Central Pontides and along the Bitlis-Zagros suture
zone), epithermal deposits of both LS and HS type (mostly in western Anatolia) and
significant number of IS type with noteworthy lead and zinc presence, and carbonate-
hosted sulfidic and nonsulfidic Pb-Zn deposits (along the Tauride Belt). In addition,
skarn-type (mainly Fe producing) deposits (disseminated throughout the country),
porphyry Cu-Mo (disseminated throughout the country), mesothermal cupriferous Pb-
Zn mineralizations (along the Eastern Pontide belt), and podiform chromite deposits
(along the suture zones throughout the country) are also important part of the metal-
logeny of Turkey. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 84–89, 2019. https://doi.org/10.1007/978-3-030-22974-0_19 Ore Mineralogy of Kirazliyayla (Yenisehir-Bursa-Turkey) 85 2
Methods and Approaches A total of 30 samples representing the ore deposit from open pit – main production step
and from the boreholes were taken, from which polished sections were prepared and
Nikon Eclipse LV100 reflected light microscopy integrated with a CITL MK5
Cathodoluminescence system was employed for mineralogical examination. The ore
minerals and the paragenesis were identified on the basis of their petrographical fea-
tures and their textural relationships, respectively. Electron Probe Micro Analysis
(EPMA) and Secondary Electron Microscopy-Energy Dispersive Spectroscopy (SEM-
EDS) were routinely used for confirming the minerals and chemistry of sulfide minerals
when needed. F. Javid and E. Çiftçi 86 Fig. 1. (A) sphalerites and carbonates are partially replaced by late stage galena; (B) chalcopyrite
and pyrite are partially replaced by late stage galena; (C) galena replaces coarse sphalerite
(D) galena replaces coarse sphalerite and also carbonate gangue; (E) large cataclastic pyrite
veined by sphalerite and late stage galena; (F) large cataclastic sphalerite with replacing galena
and chalcopyrite, all in silicic matrix; (G) Large sphalerite grains with late stage galena
chalcopyrite and tennantite veins; (H) large pyrite and chalcopyrite veined by late stage galena
(I) Euhedral quartz, large sphalerite grains with late stage galena; (J) sphalerite veined by late
stage chalcopyrite and pyrite; (K-L) Late carbonates veining sphalerite (Py: pyrite; Ccp
chalcopyrite; Sp: sphalerite; Gn: galena; Tn: tennantite; Cal/Dol: calcite/dolomite) Fig. 1. (A) sphalerites and carbonates are partially replaced by late stage galena; (B) chalcopyrite
and pyrite are partially replaced by late stage galena; (C) galena replaces coarse sphalerite;
(D) galena replaces coarse sphalerite and also carbonate gangue; (E) large cataclastic pyrite
veined by sphalerite and late stage galena; (F) large cataclastic sphalerite with replacing galena
and chalcopyrite, all in silicic matrix; (G) Large sphalerite grains with late stage galena,
chalcopyrite and tennantite veins; (H) large pyrite and chalcopyrite veined by late stage galena;
(I) Euhedral quartz, large sphalerite grains with late stage galena; (J) sphalerite veined by late
stage chalcopyrite and pyrite; (K-L) Late carbonates veining sphalerite (Py: pyrite; Ccp:
chalcopyrite; Sp: sphalerite; Gn: galena; Tn: tennantite; Cal/Dol: calcite/dolomite) Ore Mineralogy of Kirazliyayla (Yenisehir-Bursa-Turkey) 87 Fig. 1. (continued) Fig. 1. (continued) Fig. 1. (continued) 3
Results and Discussion Field observations indicated that kaolinitization with silicification is pervasive alter-
ations indicating low pH hydrothermal activity resulted in the mineralization. The
mineralization is associated with andesitic volcanics in that vein/veinlets, replacement
in places, stockwork, and breccia ore structures are prevalent. Thickness of veins may
reach to 1 m in rare occasions. Samples were collected from the production steps in the
open pit and from the boreholes cutting the mineralized zones. Based on the studies of the ore samples, the major ore minerals are sphalerite and
galena as zinc and lead carrier. Pyrite is a ubiquitous. Chalcopyrite is a minor to trace F. Javid and E. Çiftçi 88 sulfide phase along with trace tennantite (Fig. 1K–L). Most of the samples indicate that
the mineralization is medium to high grade. Sphalerite and galena occur as large grains
(sometimes up to mm size) in most of the samples, indicative of being precipitated out
of supersaturated fluids within narrow spaces. Both are also fairly inclusion-free
(clean). When not, galena occurs with tennantite, sphalerite with pyrite and chal-
copyrite. Galena has also chalcopyrite encapsulation in places. Interpretation of the intergrowth ore textures suggests that mineralization started
with pyrite crystallization then a brief precipitation of first generation of chalcopyrite,
then followed by major sphalerite crystallization, which is followed by a major car-
bonatization, followed by a brief tennantite and second generation of chalcopyrite
formation. In the final stage of the ore mineralization a major galena precipitation took
place. Figure 2 summarizes the mineralization event. Ore deposit experienced very
weak supergene stage in which only traces of covellite formed. Fig. 2. Proposed ore mineral paragenetic sequence for the Kirazlıyayla Zn-Pb mineralization Fig. 2. Proposed ore mineral paragenetic sequence for the Kirazlıyayla Zn-Pb mineralization Ore Mineralogy of Kirazliyayla (Yenisehir-Bursa-Turkey) 89 Acknowledgements. The authors would like to acknowledge support received from Meyra
Mining company for their courteous support during the field work. 4
Conclusions Major ore minerals include sphalerite and galena. Chalcopyrite is minor while ten-
nantite is trace. Pyrite is always present, but not as much as sphalerite and/or galena. Tennantite is the only fahlerz that occurs in some ore zones, overall in trace quantities. Sphalerite and galena occur as large grains in most of the samples. Both are also fairly
inclusion-free (clean). When not, galena occurs with tennantite, sphalerite with pyrite
and chalcopyrite. Galena has also chalcopyrite encapsulation in places. Paragenetic
succession for the ore minerals appear to be (from early to late): Pyrite-Chalcopyrite (I)-Sphalerite-Chalcopyrite (II)-Tennantite-Galena Calcite/dolomite and quartz account for the gangue minerals. Kaolinite is the major
clay mineral. Sericite also locally become significant. Coarse nature of the major minerals (majority of the galena and sphalerite is larger
than 100 microns) suggests high liberation (>90%). Small size occurrences (<10%) are
mostly in the form of veinlets within each other, so during milling, most of that size
range may be liberated. Sulfosalts or fahlore are represented only by tennantite (Cu6[Cu4(Fe,Zn)2]As4S13) Preliminary results show that the Kirazlıyayla mineralization is a Zn-Pb (±Cu)
hosted by andesitic volcanics whose emplacement within the metamorphic complex
should be tectonically controlled. Ore Mineralogy of Kirazliyayla (Yenisehir-Bursa-Turkey) 1 Department of Mineral Technology, School of Geology,
Transbaikal State University, Chita, Russia
2 Department of Geology, University of Kazan, Kazan, Russia
nesvvik@gmail.com Abstract. We have considered problem of accumulation, storage, utilization
and recycling of wastes of various industries. We chose borogypsum as the
object of study, which contains gypsum and silicon dioxide, which can be used
in various industries. We proposed a new flotation reagent for separation of
silicon concentrate from wastes of boric acid production. Using methods of
mathematical planning we conducted a multifactorial experiment, which
allowed identifying optimal flotation mode. We developed a technology for
processing of borogypsum. Keywords: Secondary raw Borogypsum Flotation Perlastane
White soot Silicon concentrate © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 90–93, 2019.
https://doi.org/10.1007/978-3-030-22974-0_20 References Delaloye M, Bingöl E (2000) Granitoids from western and northwestern anatolia: geochemistry
and modeling of geodynamic evolution. Int Geol Rev 42:241–268 Okay AI, Tüysüz O, Satır M, Özkan-Altıner S, Altıner D, Sherlock S, Eren RH (2006)
Cretaceous and Triassic subduction-accretion, HP/ LT metamorphism and continental growth
in the Central Pontides, Turkey. Geol Soc Am Bull 118:1247–1269 Okay Aİ, Monié P (1997) Early Mesozoic subduction in the Eastern Mediterranean: evidence
from Triassic eclogite in northwest Turkey. Geology 25:595–598 Okay Aİ, Monod O, Monié P (2002) Triassic blueschists and eclogites from northwest Turkey:
vestiges of the Paleo-Tethyan subduction. Lithos 64:155–178 Topuz G, Altherr R, Schwarz WH, Dokuz A, Meyer HP (2007) Variscan amphibolite facies
metamorphic rocks from the Kurtoğlu metamorphic complex (Gümüşhane area, Eastern
Pontides, Turkey). Int J Earth Sci 96:861–873 Topuz G, Altherr R, Kalt A, Satır M, Werner O, Schwartz WH (2004) Aluminous granulites from
the Pulur Complex, NE Turkey: a case of partial melting, efficient melt extraction and
crystallisation. Lithos 72:183–207 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Use of Borogypsum as Secondary Raw A. Khatkova1, L. Nikitina2, and S. Pateyuk2(&) 1 Department of Mineral Technology, School of Geology,
Transbaikal State University, Chita, Russia
2 Department of Geology, University of Kazan, Kazan, Russia
nesvvik@gmail.com 2
Methods and Approaches A sample of waste of boric acid production, borogypsum, obtained from Dalne-
gorsky MPP was selected as the object of study. For the flotation of gypsum-containing raw, fatty acid reagents are commonly used
(Bulut et al. 2008; Matsuno et al. 1958). We proposed a new flotation reagent perlastan
ON 60 to isolate the silicon concentrate by reverse flotation. It was efficiently used at
the flotation of non-metallic fluorite ores (Dolgikh 2012), and is a promising flotation
reagent for borogypsum. To determine optimal conditions for flotation, we used the method of rational
planning of a multifactorial experiment (Malyshev 1977). 1
Introduction The dependence of the need for concentrate on the content of SiO2 According to the table, it is necessary to ensure the content of silicon dioxide from
50 to 65% with a product yield 15% and above using flotation. 1
Introduction Over the years the enterprises of mining and chemical industries in the Far East
accumulated millions tons of industrial wastes that are currently not being recycled. Thus, the total amount of wastes from the production of boric acid - borogypsum - in
the Far Eastern region is more than 25 million tons. Borogypsum contains gypsum and
silicon dioxide, which can be used in various industries. In this regard, the problem of
complex processing of these wastes to obtain various functional materials is a very
urgent task (Gordienko et al. 2014). Highly dispersed amorphous silica is referred to as white soot. White soot of all
kinds, alongside with black ones, is used to strengthen rubber, being absolutely
indispensable for silicone rubbers. It is also used as fillers in the production of rubber
linoleum, as well as in plastics, paints, lubricants and other materials to give them
valuable properties. In addition to silicon dioxide, another component of borogypsum, gypsum and
anhydrite, can be used in industry. All possible products (component for cement
production, dry building mixtures, finished building and architectural products, gypsum
boards) have positive market prospects in Russian markets and in some neighboring
foreign markets. In Russia there are projects for the production of white soot with a high silica
content by separating from datolite tailings. However, such technologies are not
implemented at industrial scale, because they are quite expensive due to the low
content of silicon dioxide in the original product. Use of Borogypsum as Secondary Raw 91 Use of Borogypsum as Secondary Raw To reduce the consumption of acids and the cost of production of white soot, it is
necessary to increase the content of silicon dioxide in the original product by other
cheaper methods such as flotation. To reduce the consumption of acids and the cost of production of white soot, it is
necessary to increase the content of silicon dioxide in the original product by other
cheaper methods such as flotation. The table below shows the dependence of the need for concentrate on the content of
silicon dioxide (Table 1). Table 1. The dependence of the need for concentrate on the content of SiO2
SiO2 content
in concentrate
Need for
concentrate,
t/hour
Need for sludge t/hour, with
a concentrate yield 15%
Need for pulp volume,
m3/h, at 20% solids
65
1,35
9
40,1
60
1,47
9,8
43,7
55
1,60
10,7
47,7
50
1,76
11,7
52,1 Table 1. 3
Results and Discussion After conducting a multifactorial experiment, the following regularities were identified. Increasing temperature of flotation leads to decreasing extraction, but at the same
time, the content of silicon dioxide increases. It was also found that the temperature
affects the floatability of gypsum, however, almost no effect on the floatability of
silicon concentrate. Consequently, by changing the temperature, it is possible to reg-
ulate the process, depending on the task. Changes in flotation pH slightly affect the
content and recovery, therefore, pH control is not effective, which in turn is an
advantage of this method, for it does not require consumption of additional reagents. Increasing concentration of perlastane increases the content of silicon concentrate in the
chamber product, however, the extraction drops significantly. The agitation time has a A. Khatkova et al. 92 linear effect on the extraction, at the same time we didn’t observe a clear dependence of
the agitation time on the content. Therefore it is necessary to agitate perlastane with
minerals for at least 3 min. As the flotation time increases, the content slightly
decreases due to the fact that alongside with calcium sulphate, silica dioxide begins to
shift to the foam product. On the basis of the revealed regularities we determined optimal conditions of
flotation. However, the content of silicon concentrate in the chamber product remains
insufficient. From the literature it was found that sodium sulfide or liquid glass was often used to
suppress flotation of quartz. Experiments with these reagents showed that they prac-
tically did not suppress flotation of amorphous silicon dioxide, and could also suppress
flotation of gypsum, therefore it would be not advisable to use them for borogypsum
flotation. We determined that it would be much more effective to increase the concentration
of perlastane at the main flotation, increasing its time than to feed the reagent frac-
tionally in different operations. Increasing the concentration of perlastane up to 1 kg/t,
as well as flotation time up to 9 min, allows obtaining a product with a content of
53.6%, with a yield 32.5% and a recovery 64%. 4
Conclusions We suggested a new technology for processing wastes of boric acid production
according to the scheme presented in Fig. 1. The proposed scheme has its advantages
in comparison to previously known technologies. Thus it does not require regulation of
pH of the medium, consists of a single operation, which significantly reduces flotation
time and also allows to increase the yield of the chamber product significantly. Borogypsum pulp 20%
1 kg/t ON-60 3 min agitation
Reverse flotation 9 min (T=300C, pH=7.5)
Foam product
(gypsum sludges)
Chamber product
(silica concentrate
CSiO2=53.5%)
Fig. 1. Diagram of borogypsum flotation Borogypsum pulp 20% Chamber product
(silica concentrate
CSiO2=53.5%) Foam product
(gypsum sludges) Fig. 1. Diagram of borogypsum flotation Use of Borogypsum as Secondary Raw 93 New Approaches in X-ray Phase Analysis
of Gypsum Raw Material of Diverse Genesis
V. Klimenko(&), V. Pavlenko, and T. Klimenko
Belgorod State Technological University named after V G Shukhov,
Belgorod, Russia
klimenko3497@yandex.ru V. Klimenko(&), V. Pavlenko, and T. Klimenko Belgorod State Technological University named after V G Shukhov,
Belgorod, Russia
klimenko3497@yandex.ru Abstract. Modern software was used to conduct X-ray phase analysis of
gypsum raw material of different genesis: gypsum from several deposits, citr-
ogypsum, phosphogypsum, vitamin gypsum, chemically pure gypsum, hydra-
tion products from different gypsum and anhydrite cementing and composite
materials on their bases. Two criteria for ranging gypsum raw material with the
account of genesis and structural characteristics and predicting properties of
gypsum bindings and materials based on them were suggested: structure
sophistication criteria (Кg) and identity criteria (Кi). They were calculated by the
results of X-ray phase analysis of calcium sulphate. Keywords: Gypsum X-ray phase analysis Structure sophistication criteria
Identity criteria Gypsum texture and structure References Bulut G, Atak S, Tuncer E (2008) Celestite-gypsum separation by flotation. Can J Metall Mater
Sci 47(2):119–126 Bulut G, Atak S, Tuncer E (2008) Celestite-gypsum separation by flotation. Can J Metall Mater
Sci 47(2):119–126 Dolgikh OL (2012) The use of perlastane reagent as an alternative to oleic acid during fluorite
flotation. Bull ZabGU 9(88):20–26 Gordienko PS, Kozin AV, Yarusova, SB, Zgibly IG (2014) Integrated processing of waste from
the production of boric acid to produce materials for the construction industry. Min Inf Anal
Bull (S4-9):60–66 Gordienko PS, Kozin AV, Yarusova, SB, Zgibly IG (2014) Integrated processing of waste from
the production of boric acid to produce materials for the construction industry. Min Inf Anal
Bull (S4-9):60–66 Malyshev VP (1977) Mathematical planning of metallurgical and chemical experiments. Alma-
Ata: Sci 35 Malyshev VP (1977) Mathematical planning of metallurgical and chemical experiments. Alma-
Ata: Sci 35 Matsuno T, Kadota M, Ishiguro Y (1958) Separation of gypsum by the flotation process. Bull
Soc Salt Sci 12(2):73–78 Matsuno T, Kadota M, Ishiguro Y (1958) Separation of gypsum by the flotation process. Bull
Soc Salt Sci 12(2):73–78 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction There are convincing theoretical and experimental data about physical-chemical pro-
cesses of gypsum dehydration, their structural defects dominating role, an origin and
transformation mechanism of these defects during calcinations, at the same time there
are no data about influence of the gypsum raw material formation on these processes. Thermodynamic characteristics of gypsum with different structure are rather close
(Kelly et al. 1948; Reference… 2004) and it is difficult to study gypsum raw material
deformations according to them. There are data (Gordashevski 1963) about application
of differential thermal and x-ray analysis methods for these purposes. To characterize
gypsum from different deposits P.F. Gordashevskiy (Gordashevski and Sakhno 1963)
suggested identity criteria – the value received by division of reflexes intensivities
difference at 2H = 20o and 23o by their sum. On the other hand, gypsum crystal lattice
defects can be characterized by crystallinity degree, which is determined by reflex with
d = 2.81 Å in the X-ray patterns (Boldyrev 1983). Crystallinity degree diagnostics is
possible in relation to diffraction reflection doublet intensivity and by crystallinity
index determination. Some authors suggest studying crystal lattice defects by studying
X-ray diffraction line broadening (Ginje 1961). Literature analysis shows wide
researches in this sphere as well as absence of practical standards. In connection with
this the purpose of this work was to find new criteria allowing ranging gypsum raw
material with the account of genesis and structural characteristics at designing com-
posite materials. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 94–98, 2019. https://doi.org/10.1007/978-3-030-22974-0_21 New Approaches in X-ray Phase Analysis of Gypsum Raw Material 95 2
Materials and Methods As raw materials we studied gypsum of Shedok, Baskunchak, Novomoskovsk, and
Peshelansk Deposits; man-induced gypsum – chemically pure gypsum, citrogypsum,
vitamin gypsum, Voskresensk phosphogypsum, synthetical gypsum; gypsum thermal
treatment and rehydration products, composite materials based on gypsum binders,
multiphase gypsum binders (MGB). To receive composite materials, we used crashed
glass withdrawals (CGW) and iron-ore concentrate of Lebedinsk mining and pro-
cessing enterprise (OMC). X-ray phase analysis of the studied species was done with
X-ray diffraction meter DRON-4 by powder pattern method. Grain-size analysis of
powdery material was done by laser granulometry method with a MicroSizer 201
installation. 3
Results and Discussions The X-ray phase analysis of the initial raw material showed that intensity and area of
the main reflexes depend on its genesis. Intensity (L) and reflex area (S) at interplanar
spacing (d) 7,628 Å are strongly dependent on gypsum raw material. Gypsum raw
material genesis influences somewhat lower the intensity and reflex area at d = 4.291
and 3.069 Å. Reflex intensity at d = 3.809 Å almost independent of the gypsum raw
material nature. The received data evidence that the studied gypsum samples have
equal crystal lattice dimensions. To determine the gypsum raw material genesis influence on its structure and crystal
lattice defects several studies of the initial gypsum raw material crystallinity degree and
its products of the heat processing have been studied. Here indexes were suggested
calculated by reflex area difference division by their sum. The calculations were done
for reflexes with the most intensity such as: 7.628 Å; 4.291 Å; 3.809 Å; 3.069 Å; 2.880
Å; 2.687 Å. We found that most indexes slightly depended on raw material genesis. Certain
dependencies were observed in changes of two indexes chosen for further work. The
first index was structure sophistication index (Кg), and the second was identity index
(Кi). Кg, was determined by X-ray phase analysis results, as ratio of reflex area dif-
ference division at 2H = 29.09° (d = 3.069 Å) and 2H = 31.08° (d = 2.876 Å) by
their sum, a Кi as a ration of reflex area difference division at 2H = 20.68° (d = 4.291
Å) and 2H = 31.08° (d = 2.876 Å) by their sum. Kg ¼ S29:09 S31:08
S29:09 þ S31:08
Ki ¼ S20:68 S31:08
S20:68 þ S31:08 Kg ¼ S29:09 S31:08
S29:09 þ S31:08
Ki ¼ S20:68 S31:08
S20:68 þ S31:08 For natural gypsum Кg = 0.42–0.46, and for production induced gypsum was
0.560–0.903 (Table 1). The greater the value Кg, the more sophisticated crystal
structure was and gypsum raw material was less stable. We think that Кi value depends on gypsum raw material micro-assembly dimen-
sions. Powdery-material grain-size composition determined by laser granulometry
analysis proves a finely crystalline structure of phosphogypsum and citrogypsum. V. Klimenko et al. 96 Table 1. X-ray phase analysis of gypsum of different genesis Table 1. 4
Conclusion A method and parameters (structure sophistication index (Кg) have been suggested and
identity index (Кi)) allowing ranging gypsum raw material with the account of genesis
and structural characteristics and forecast properties of gypsum cementing components
and materials on their basis. It has been determined that for natural gypsum Кg = 0.42–
0.46 and for produced gypsum Кg = 0.560–0.903. Value Кi depends on micro clusters
sizes of gypsum raw material structure. For large grained gypsum Кi = 0.612–0.746. for finely crystalline gypsums – 0.312–0.488. At producing gypsum cementing agents
it is necessary to observe that their Кg is closer to value Кg of natural gypsum and Кi is
in the range 0.38–0.41. Acknowledgements. The work is realized within the framework of the Program of flagship
university development on the base of the Belgorod State Technological University named after
V.G. Shukhov. using equipment of High Technology Center at BSTU named after V.G. Shukhov. 3
Results and Discussions 97 New Approaches in X-ray Phase Analysis of Gypsum Raw Material New Approaches in X-ray Phase Analysis of Gypsum Raw Material 97 3
Results and Discussions X-ray phase analysis of gypsum of different genesis
№
Gypsum genesis
Index value
Ki
Kg
1
Chemically pure gypsum
0.612
0.660
2
Shedok deposit gypsum
0.362
0.440
3
Baskunchak deposit gypsum
0.448
0.420
4
Novomoskovsk deposit gypsum
0.448
0.450
5
Peshelansk deposit gypsum
0.215
0.450
6
Vitamin gypsum
0.746
0.903
7
Voscresensk phosphogypsum
0.404
0.560
8
Citrogypsum 1
0.312
0.640
9
CaCl2 and (NH4)2SO4 gypsum
-
0.749
10
Anhydrite cement (1,5% (NH4)2SO4 + 0,5%CuSO4)
0.277
0.169
11
Composite material based on MGB and CGW remains (Klimenko
et al. 2013)
0.286
0.156
12
Composite material: 70% CuSO4 II + 30% LC
70% Г-4 + 30% LC (Klimenko et al. 2018)
0.370
0.270
-
- Crystal size of production induced chemically pure gypsum and vitamin gypsum is
higher. They can be referred to large grained gypsum. By value Кi Shedok natural
gypsum can be referred to finely crystalline raw material (finely tessellated oriented
structure). Baskunchak natural gypsum has medium tessellated chaotic structure, and
according to value Кi it takes intermediate position. Hence, for large grained gypsum Кi = 0.612–0.746, for finely crystalline gypsum
Кi = 0.312–0.488. Granulometry of produced gypsum changes unevenly. There are
fractions, which number depends greater on genesis, and there are fractions, which
number depends less on raw material genesis. To analyze the influence of gypsum heat
treatment parameters and amount of residue hydrate water on value Кg and Кi we used
Baskunchak natural gypsum. The results show that Кg of calcium sulphate hydration
products can be both greater and less Кg of natural gypsum. Gypsum produced during
heat treatment products hydration with hydrate water 11.00–14.42 mass%, has
sophistication structure index value greater than that of natural gypsum. The smallest
value Кg, irrespective of heat treatment parameters is of calcium sulphate hydration
products with hydrate water amount 3–4 mass%. Calcium sulphate hydration products
having value Кg close to Кg of the initial gypsum, have maximum strength. The amount
of residue hydrate water of these products is equal 1.0–1.5 mas. %. By value Кi (0.39–
0.45) the produced gypsum can be classified as fine- or medium crystalline structure. Analysis of identity index value suggests that calcium sulphate hydration products
with the amount of hydration water 10–15 mass% have bigger crystal micro cluster size
and hydration products b-CaSO40.5H2O. b-centrifuged hemihydrates of calcium
sulphate and soluble anhydrous plaster (b-CaSO4III) have smaller size of gypsum
crystal micro clusters (Кi = 0.35–0.43). 1
Introduction The Republic of Uzbekistan is one of the leading gold mining countries in the world. Such large deposits as Muruntau, Myutenbay, Kokpatas, Zarmitan and others
(Kyzylkum-Nurata ore region) are known. As a result of many years of production, the
mining and processing industry faces the problem of transition from oxidized ores with
large “free” gold to the processing of refractory ores with “invisible” gold. One of the
effective approaches to the assessment of such ores is the use of nanotechnology,
nanomineralogy and nanogeochemistry (Hochella 2002; Koneev et al. 2010). © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 99–102, 2019.
https://doi.org/10.1007/978-3-030-22974-0_22 Nanotechnologies in Mineral-Geochemical
Methods for Assessing the Forms of Finding
of Gold, Related Elements, Technological
Properties of Industrial Ores and Their Tails R. Koneev1(&), R. Khalmatov2, O. Tursunkulov2, A. Krivosheeva1,
N. Iskandarov2, and A. Sigida1 R. Koneev1(&), R. Khalmatov2, O. Tursunkulov2, A. Krivosheeva1,
N. Iskandarov2, and A. Sigida1 1 National University of Uzbekistan, Tashkent, Uzbekistan
ri.koneev@gmail.com
2 Centre for Advanced Technology, Tashkent, Uzbekistan 1 National University of Uzbekistan, Tashkent, Uzbekistan
ri.koneev@gmail.com
2 Centre for Advanced Technology, Tashkent, Uzbekistan Abstract. Seven mineralogical-geochemical types of ores have been identified
in the gold orogenic deposits of Uzbekistan: /Au-W/Au-Bi-Te/Au-As/Au-Ag-
Te/Au-Ag-Se/Au-Sb/Au-Hg/. Non-industrial are Au-W and Au-Hg. For each
industrial type, certain gold compounds are characteristic: maldonite, Au-
arsenopyrite and Au-pyrite, petzite, physhesserite, petrovskaite, aurostibite,
which form regular micro- nanoensembles with the corresponding minerals Bi,
Te, Se, S, As, Sb. They are direct indicators of the type and technological
properties of ores. Pyrite and arsenopyrite are preserved in the processing wastes
of Au-Bi-Te and Au-As ores, in which gold (901‰), maldonite (Au2Bi),
headleyite (Bi7Te3) and others were detected, 300–700 nm in size. Waste is
suitable for the secondary extraction of gold. Keywords: Nanomineralogy Gold Ores Tails Properties Technologies Keywords: Nanomineralogy Gold Ores Tails Properties Technologies References Boldyrev VV (1983) Experimental methods in mechanochemistry of solid inorganic matters. Science, Novosibirsk, 36 p Ginje A (1961) Crystal X-ray filming. State Publishing House of Physic-Mathematical Literature,
Moscow, 188 p Gordashevskiy PF, Sakhno ZA (1963) About some properties of gypsum raw material of
different crystalline structure, no 26. Collection of works/SRILCM, Moscow, pp 25–27 Gordashevskiy PF (1963) Thermal and X-ray phase gypsum analyses results. Constr Mater
(12):28–30 Kelly N, Suttard D, Anderson К (1948) Thermodynamic properties of gypsum. BTI MCMPI,
Moscow, pp 38–42 Klimenko VG, Kashin GA, Prikaznova TA (2018) Plaster-based magnetite composite materials
in construction. In: IOP Conference Series: Materials Science and Engineering, vol 327,
p 032029. https://doi.org/10.1088/1757-899x/327/3/032029 Klimenko VG, Pavlenko VI, Gasanov SK (2013) The role of pH medium in forming binding
substauces on base of calcium sulphate. Middle-East J Sci Res 17(8):1169–1175 Reference book. Gypsum materials and articles (production and application) (2004) Under
general editorship of professor, doctor of sciences A.V. Ferronskaya. Construction
Universities Association Publishing House, Moscow, 485 p V. Klimenko et al. 98 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches The section of mineralogy that studies the conditions of formation, the physicochemical
properties of natural compounds, the size of which, at least in one dimension, enters the
nanoscale (10−6–10−9 m) is called nanomineralogy. There are nanominerals that are
formed by the “down-top” technology, from atoms to chemical compounds, and R. Koneev et al. 100 nanoparticles that are formed by the “top-down” technology during dispersion. The
anomalous properties of nanominerals and nanoparticles are determined by a signifi-
cant increase in the specific surface energy. As a result of “size effects”, nano-objects
acquire high chemical, catalytic, sorption activity and other properties. “Noble”,
chemically inert gold in the nanostate becomes active and is found in ores not only in
the form of native, but also in the form of a compound with various elements up to
sulfur and oxygen. When studying ores, the main focus is on ore minerals. Non-
metallic minerals, carbonaceous material rather negatively affect the enrichment pro-
cesses, making it difficult to crush, float or adsorb gold. The study of mineralogy and
nanomineralogy is important due to the fact that minerals of different composition and
size differ in their ability to crush, float, or cyanate. The studies used an electron probe
microanalyzer Superprobe JXA-8800R, Carl Zeise Oxford instrument (SEM-EDX);
ICP MS 7500 Agilent Technologies. 3
Results and Discussion The gold ore deposits of the Kuzylkum-Nurata region of Uzbekistan are confined to the
South Tien Shan orogenic belt. Placed in the “black” shale, terrigenous, carbonate,
volcanogenic and intrusive rocks. Tails of ore enrichment stored in long-term facilities
were also studied. It has been established that in the orogenic deposits of the
Kyzylkum-Nurata region, seven mineral-geochemical ore types are developed: /Au-
W/Au-Bi-Te/Au-As/Au-Ag-Te/Au-Ag-Se/Au-Sb/Au-Hg/. Each type differs in the ratio
of non-metallic and ore (sulphide) minerals, but primarily in the composition of micro-
nanoensembles of minerals, compounds and gold fineness. In the studied deposits, the
leading industrial ones are Au-Bi-Te, Au-As, Au-Ag-Se, Au-Sb (Table 1). In addition
to native gold, electrum and kustelite, maldonite, Au-arsenopyrite, Au-pyrite, petzite, able 1. Types, composition of ores of gold and gold-silver deposits of Kyzylkum-Nurat
gion Table 1. Types, composition of ores of gold and gold-silver deposits of Kyzylkum-Nurata
region Nanotechnologies in Mineral-Geochemical Methods 101 Fig. 1. Micro-nanomineral forms of gold release and its compounds in various types of ores
(I) and tails (II). a - maldonite, Bi-tellurides in arsenopyrite; b - gold, arsenopyrite in arsenic
pyrite; c - aurostibite with arsenopyrite in antimonite; d - nanogold in pyrite; e - maldonite and
hedleyite and arsenopyrite; f - hedleyite in arsenopyrite. Fig. 1. Micro-nanomineral forms of gold release and its compounds in various types of ores
(I) and tails (II). a - maldonite, Bi-tellurides in arsenopyrite; b - gold, arsenopyrite in arsenic
pyrite; c - aurostibite with arsenopyrite in antimonite; d - nanogold in pyrite; e - maldonite and
hedleyite and arsenopyrite; f - hedleyite in arsenopyrite. calavertite, fishesserite, petrovskaite, aurostibite were detected in ores. Gold release
forms and its micro- nanoanesmble in ores in Fig. 1, row I. The study of Au-Bi-Te and
Au-As ore tails showed that some of the sulfide minerals still remain, in which the
nanoforms of gold, its compounds and associated bismuth tellurides are preserved 300–
700 nm in size (Fig. 1, row II).nts. Acknowledgements. The authors are grateful to the Navoi Mining and Metallurgical Combi-
nate for their assistance in research. 4
Conclusions As a result of the research the following were established: - Seven mineral-geochemical
types of ores were identified in the orogenic deposits of the Kyzylkum-Nurata region of
Uzbekistan: /Au-W/Au-Bi-Te/Au-As/Au-Ag-Te/Au-Ag-Se/Au-Sb/Au-Hg/. Industrial
types are /Au-Bi-Te/Au-As/Au-Ag-Se/Au-Sb/ ores possessing different technological
properties; - Gold is represented by its native type, electrum, cyustelite, maldonite,
petzite, aurostibite, fishesserite, petrovskaite, Au-arsenopyrite, Au-pyrite, forming
micro- nanoensembles with the Bi, Te, Sb, Se, As, S minerals; - Pyrite and arsenopyrite
remain in the enrichment tails of Au-Bi-Te and Au-As ores, in which nanoinclusions of
gold, maldonite and bismuth tellurides are found. Tails are recyclable; - Nanotech-
nological methods allow to increase the efficiency and unambiguity of exploration,
assess the prospects for hidden and unconventional deposits, as well as assess the
technological properties of ores and enrichment tails for their complete processing. R. Koneev et al. 102 Hochella M (2002) There’s plenty of room at the bottom: nano-scince in geochemistry. Geochem
Cosmochem Acta 66(5):735–743 Hochella M (2002) There’s plenty of room at the bottom: nano-scince in geochemistry. Geochem
Cosmochem Acta 66(5):735–743
Koneev RI, Khalmatov RA, Mun YS (2010) Nanomineralogy and nanogeochemistry of ores Koneev RI, Khalmatov RA, Mun YS (2010) Nanomineralogy and nanogeochemistry of ores
from gold deposits of Uzbekistan. Geol Ore Deposits 52(8):755–766 Hochella M (2002) There’s plenty of room at the bottom: nano-scince in geochemistry. Geochem
Cosmochem Acta 66(5):735–743
Koneev RI, Khalmatov RA, Mun YS (2010) Nanomineralogy and nanogeochemistry of ores
from gold deposits of Uzbekistan. Geol Ore Deposits 52(8):755–766 Applied Mineralogy of Mining
Industrial Wastes O. Kotova1(&) and E. Ozhogina2 1 Institute of Geology Komi SC UB RAS, Syktyvkar, Russia
kotova@geo.komisc.ru
2 FSBE “All-Russian Institute of Mineral Raw Materials” (VIMS),
Moscow, Russia 1 Institute of Geology Komi SC UB RAS, Syktyvkar, Russia
kotova@geo.komisc.ru
2 FSBE “All-Russian Institute of Mineral Raw Materials” (VIMS),
Moscow, Russia Abstract. Recycling and disposal of mining wastes has been very important
and considered as an independent task. Features of composition and structure of
mining wastes, identified by a set of mineralogical analysis methods, allowed
predicting their possible involvement in the secondary processing. This was
illustrated by the example of metallurgical iron slags. Keywords: Mining wastes Technological mineralogy Iron slags References Hochella M (2002) There’s plenty of room at the bottom: nano-scince in geochemistry. Geochem
Cosmochem Acta 66(5):735–743 Koneev RI, Khalmatov RA, Mun YS (2010) Nanomineralogy and nanogeochemistry of ores
from gold deposits of Uzbekistan. Geol Ore Deposits 52(8):755–766 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 103–106, 2019.
https://doi.org/10.1007/978-3-030-22974-0_23 1
Introduction The intense development of the mineral resource complex inevitably leads to accu-
mulation of significant amounts of waste, which negatively affect natural ecosystems. Therefore, the disposal and recycling of wastes is important at the national level and
considered within framework of the priority direction of development of science and
technology in the Russian Federation – “Rational environmental management”. Today
we can state the fact that recycling is an independent major task for our industry. Mining and processing wastes are very diverse. They include overburden, host
rocks, dry raw processing, off-balance sheet and non-standard ores, which composition
and properties are not only close to their natural analogues, but usually used the same
way. Processing wastes of metallurgical, chemical, heat power industries are more
abundant. Slags, muds, ash slags and oil muds, burnt rocks, pyrite cinders, clinkers,
dusts are significantly different from natural ores and rocks. They are characterized by
variable granular composition, often a high dispersion, presence of amorphous for-
mations, complex interrelationship of mineral and (or) technogenic phases, including
presence of eutectic colonies or structures of decomposition of solid solutions, a small
amount of one or more minerals, polymineral (polyphase) aggregates, presence of
isomorphic minerals and polytypic modifications, secondary changes associated mainly
with hypergenesis. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 103–106, 2019. https://doi.org/10.1007/978-3-030-22974-0_23 O. Kotova and E. Ozhogina 104 2
Methods and Approaches The study of the composition, structure and technological properties of mining wastes
is based on modern scientific, methodological, technical and instrumental support for
the researches of technogenic raw and allows predicting its possible involvement in
secondary processing, including elimination of environmental consequences of
industrial processing (Ozhogina et al. 2018; Ozhogina et al. 2017; Chanturia et al. 2016; Yakushina et al. 2015; Ozhogna and Kotova 2019; Burtsev et al. 2018). Necessary and sufficient mineralogical information about the object is possible to get
by a complex of mineralogical and analytical methods (optical and electron micro-
scopy, X-ray, X-ray tomography, micro X-ray spectral analysis). For different types of
wastes, an individual set of mineralogical analysis methods is used, which allows to
obtain complete and reliable information, including information on the phase com-
position of technogenic formations, as well as the form of finding useful elements,
granular composition, morphometric parameters, nature of localization of specific
phases (Ozhogina et al. 2017). Mineralogical study of mining wastes is carried out mainly according to the
methodological documents developed for the analysis of natural mineral raw. Special
methods of mineralogical analysis of industrial wastes do not exist. The study of such
objects is of an interdisciplinary nature because of a reasonable combination of
methods from various fields of science and adapted to solving mineralogical, techno-
logical and environmental problems. 3
Results and Discussion The bulk of the processing wastes are slags. For example, iron-containing metallurgical
slag is a loose material with dense lumpy aggregates and few octahedral crystals. More
than 80% of the slag is represented by a thin material with a particle size of less than
0.2 mm. The main beneficial elements are iron (42.5%) and chromium (3.15%), which
form their own mineral phases. Nickel (0.4%), associated with trevorite, and cobalt
(0.08%), with unknown occurrence form, can possess industrial value. p
A significant part of the slags (more than 75%) is formed by black magnetic
material prone to artificial segregation, represented by spinelides forming a continuous
isomorphic series spinel-magnetite-chromite. The main mineral is magnetite, which
isolations are always non-uniform and contain numerous inclusions of non-metallic
phases represented by olivine, pyroxene, mica, corundum, feldspar and glass (Fig. 1a). Sometimes the grains are surrounded by a fairly flat border of iron hydroxides and
contain veinlets that do not extend beyond their boundaries (Fig. 1b). Two varieties of
magnetite are noted; eutectic colonies are evidence of their simultaneous presence. This
may be due not only to the closest intergrowth of the phases, or the oxidation process of
a magnet and its partial transition to maghemite, but also continuous isomorphic
substitutions of the ferrospinels, including heterogeneous −2 and 3 valence cations. Applied Mineralogy of Mining Industrial Wastes 105 Fig. 1. A. Magnetite (light gray) with inclusions of non-metallic minerals, in the center of the
grain is maghemite (white). B. Magnetite (light gray) with veins and border of iron hydroxides. Polished thin section, nicols are parallel Fig. 1. A. Magnetite (light gray) with inclusions of non-metallic minerals, in the center of the
grain is maghemite (white). B. Magnetite (light gray) with veins and border of iron hydroxides. Polished thin section, nicols are parallel The features of the mineral composition and structure of iron-containing slags
(phase composition, distribution of ore phases and aggregates, their morphostructural
characteristics, heterogeneous structure of magnetite), identified by a complex of
mineralogical analysis methods, suggest the expediency of chemical processing
methods at recycling (Yakushina et al. 2015). 4
Conclusions Mining wastes, being main types of technogenic raw, possess mineralogical charac-
teristics (mineral and (or) phase composition, useful component occurrence form,
morphostructural features and distribution pattern, real composition and structure) that
determine the strategy and tactics of its secondary use: 1) as initial raw without processing, for example, for extraction of valuable metal (1) as initial raw without processing, for example, for extraction of valuable metals;
(2) as initial raw after additional processing to obtain material resources in the
industry. (2) as initial raw after additional processing to obtain material resources in the
industry. (2) as initial raw after additional processing to obtain material resources in the
industry. (3) as object of disposal. (3) as object of disposal. Ozhogina EG, Kotova OB, Yakushina OA (2018) Mining wastes: mineralogical features.
Vestnik of IG Komi SC UB RAS, pp 43–49 (in Russian) Chanturia VA, Ozhogina EG, Shadrunova IV (2016) Tasks of ecological mineralogy during the
development of the Earth’s interior. Phys Tech Probl Dev Miner Resour (SB RAS) 5:193–196
(in Russian) Yakushina OA, Ozhogina EG, Khozyainov MS (2015) Microtomography of technogenic mineral
raw materials. IG Komi Science Center, Ural Branch of the Russian Academy of Sciences.
vol 11, pp 38–43 (in Russian) V. Maslennikov1(&), S. Maslennikova1, N. Aupova1, A. Tseluyko1,
R. Large2, L. Danyushevsky2, and U. Yatimov1 1 Institute of Mineralogy, Ural Branch of RAS, Miass, Russia
mas@mineralogy.ru
2 CODES ARC Centre of Excellence in Ore Deposits, University of Tasmania,
Hobart, Australia 1 Institute of Mineralogy, Ural Branch of RAS, Miass, Russia
mas@mineralogy.ru
2 CODES ARC Centre of Excellence in Ore Deposits, University of Tasmania,
Hobart, Australia Abstract. LA-ICP-MS data on trace element zonation reflecting a local vari-
ation of physicochemical conditions of mineralization in black, grey and clear
smokers from the Pacific and Atlantic oceans are used for comparison with the
ancient chimneys of the Urals, Rudniy Altai, Pontides and Hokuroko massive
sulfide deposits. Host rocks also influence on high-tech trace element assem-
blages in chalcopyrite: ultramafic (high Se, Sn, Co, Ni, Ag and Au) ! mafic
(high Co, Se, Mo and low Bi, Au and Pb) ! bimodal mafic (high Te, Au, Ag,
Bi, Pb, Co, moderate Se, and variable As and Sb) ! bimodal felsic (high As,
Sb, Mo, Pb, moderate Bi, and low Co, Te and Se). In sphalerite of the same
range, the contents of Bi, Ga, Pb, In, Ag, Au and Sb increase versus Fe, Se, Sn
and Co. The variations in pyrite coincide with these changes. Diagenetic evo-
lution of high-tech elements is recognized in sulfide nodules. Keywords: LAICP-MS Trace elements Chimneys
Massive sulfide deposits Keywords: LAICP-MS Trace elements Chimneys
Massive sulfide deposits © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 107–110, 2019.
https://doi.org/10.1007/978-3-030-22974-0_24 References Burtsev IN, Kotova OB, Kuzmin DV, Mashin DO, Perovsky IA, Ponaryadov AV, Shushkov DA
(2018) The role of technological research in the development of the mineral resource complex
of the Timan-North Ural region. Explor Prot Miner Resour 5:38–47 (in Russian) Burtsev IN, Kotova OB, Kuzmin DV, Mashin DO, Perovsky IA, Ponaryadov AV, Shushkov DA
(2018) The role of technological research in the development of the mineral resource complex
of the Timan-North Ural region. Explor Prot Miner Resour 5:38–47 (in Russian) Chanturia VA, Ozhogina EG, Shadrunova IV (2016) Tasks of ecological mineralogy during the
development of the Earth’s interior. Phys Tech Probl Dev Miner Resour (SB RAS) 5:193–196
(in Russian) Ozhogina EG, Kotova OB, Yakushina OA (2018) Mining wastes: mineralogical features. Vestnik of IG Komi SC UB RAS, pp 43–49 (in Russian) O. Kotova and E. Ozhogina 106 Ozhogina EG, Shadrunova IV, Chekushina TV (2017) The role of mineralogical studies in
solving environmental problems of mining areas. Min J 11:105–110 (in Russian) Ozhogina E, Kotova O (2019) New methods of mineral processing and technology for the
progress of sustainability in complex ore treatment. In: 29th International Mineral Processing
Congress IMPC 2018, Canadian Institute of Mining, Metallurgy and Petroleum, 2-s2.0-
85059377649 Yakushina OA, Ozhogina EG, Khozyainov MS (2015) Microtomography of technogenic mineral
raw materials. IG Komi Science Center, Ural Branch of the Russian Academy of Sciences. vol 11, pp 38–43 (in Russian) Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches In this paper, we use new LA-ICP-MS data on minor and trace elements in sulfides
from Rudniy Altai, Pontides and Hokuroko massive sulfide deposits. Original LA-ICP-
MS data on smokers from the Pacific and Atlantic oceans are used for comparison with
the ancient chimney material. The sulfide nodules are the other important subject of the
research reflecting chemical evolution of diagenetic and metamorphic processes. Quantitative LA-ICP-MS analysis of sulfides from chimneys for a wide range of
major and trace elements (Fe, Cu, Zn, Co, Ni, Au, Ag, Bi, Pb, Tl, Cd, As, Te, Se, Mo,
Sn, V, Ti and Mn) was carried out using a New Wave 213 nm solid-state laser
microprobes coupled to an Agilent 4500 quadrupole ICP-MS housed at the CODES
LA-ICP-MS analytical facility, University of Tasmania and Institute of Mineralogy UB
RAS. For this study, quantitative analyses were performed by ablating spots ranging in
diameter from 40 to 60 microns. Data reduction was undertaken according to standard
methods (Danyushevsky et al. 2011). The LA-ICP-MS was also used for detection of
rare mineral microinclusions in the sulfides studied and for analysis of trace elements in
telluride phases. Each zone of the chimney and nodules are characterized by LA-ICP-
MS-images (Large et al. 2009). 1
Introduction The bulk minor and trace element analyses indicate significant influence of host rocks
on the metal inventory of the modern and ancient massive sulfide deposits. However,
these general data can’t provide dramatic improvement of the extraction of high-tech
elements from massive sulphide ores. In contrast, the results of the quantitative laser
ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) show concen-
tration of most trace metals in pyrite, chalcopyrite and sphalerite varieties as a function
of physicochemical parameters of the fluid as well as host rock composition. Local
LA-ICP-MS analyses of the sulphides provides a new insight into the processes
of ore treatment and recovery of high-tech elements. The chimney fragments, sulfide
turbidites, and their ore diagenites are suitable subject of this style of the research. The study is a key basement for fither improvement of mining and metallurgical
processes. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 107–110, 2019. https://doi.org/10.1007/978-3-030-22974-0_24 V. Maslennikov et al. 108 Aknowledgements. LA-ICP-MS analyses were carried out during visiting programs (2005,
2009, 2012, 2013, 2015) sponsored by the ARC Centre of Excellence grant to CODES. The
research of sulphide nodules was supported by the Russian Foundation for Basic Research
(project no. 17-05-00854) in the Institute of Mineralogy UB RAS. High-Tech Elements in Minerals of Massive Sulfide Deposits The next stage of mineral and chemical differentiation is halmyrolysis of clastic
sulfide sediments followed by diagenesis, anadiagenesis and metamorphism. Halmy-
rolysis as a leaching process of clastic sulfide sediments give way to redeposition of
high-tech element such as Se, Te, Au, Ag, Sn, and In as own minerals in enrichment
submarine supergene zones. The inclusions of authigenic mineral such as Pb, Ag, Au-
tellurides, Ag and Pb-selenides, In, Cu, Ag, Sn, Ge-sulfides, and Sn-oxide are increased
in sizes in the enrichment submarine supergene zones of massive sulfide deposits. In many massive sulfide deposits, sulphide nodules have a zonal structure: the
nucleus of the poikilite pyrite (zone A) is successively surrounded by the fringes of
metacrystalline pyrite (zone B) and marcasite (zone C). Each zone is characterized by
own mineralogical features, which are reflected in the results of LA-ICP-MS-images
(“microtopochemistry”) of the surface of the nodule cut. It is assumed that at the first
stage, the source of the substance for nodule growth was the products of the halmy-
rolysis of hyaloclastites containing an admixture of sulphide material. In the diagenetic
nucleus of nodule, the chemical elements typical of illite (Si, Al, K, Mg, V, Cr,), rutile
(Ti, W), apatite (Ca, Mn, U), galena (Pb, Bi, Sb, Ag), bismuth sulphosalts (Bi, Cu, Pb),
bornite (Cu, Bi, Ag), tetrahedrite-tennantite (Cu, As, Sb), chalcopyrite (Se, Te, Cu),
native gold (Au, Ag, Hg) and barite poikilites. In the pyrite, the high contents of Co and
Ni are suggested to be substituted for Fe2+. Only signs of tellurides of bismuth are
guessed. Zone B, probably formed in the early anadiagenetic stage, is depleted of most
trace elements, with the exception of Cu, Pb, and Ag. Formation of latest zone C was
accompanied by of Cd, In, Tl, As, Sb, Mo, and Ni saturation. 3
Results and Discussion A variety of very well preserved modern black, grey, white and clear smokers (Pacific
and Atlantic oceans) and fossil vent chimneys from the Urals, Rudniy Altai, Pontides
and Hokuroko massive sulfide deposit show systematic trace element distribution
patterns across chimneys. Chalcopyrite is enriched in Se, Sn, Bi, Co, Mo, and Te. Sphalerite in the conduits and the outer chimney wall contains elevated Sb, As, Pb, Co,
Mn, U, and V. He highest concentrations of most trace elements are found in colloform
pyrite within the outer wall of the chimneys, and likely result from rapid precipitation
in high temperature-gradient conditions. The trace element concentration in the outer
wall colloform pyrite decrease in the following order, from the outer wall inwards;
Tl > Ag > Ni > Mn > Co > As > Mo > Pb > Ba > V > Te > Sb > U > Au > Se >
Sn > Bi, governed by the strong temperature gradient. In contrast, pyrite in the high- to
mid-temperature central conduits exhibit concentration of Se, Sn, Bi, Te, and Au. The
zone between the inner conduit and outer wall is characterised by recrystallization of
colloform pyrite to euhedral pyrite, which becomes depleted in all trace elements
except Co, As and Se. The mineralogical and trace element variations between
chimneys are likely due to increasing fO2 and decreasing temperature caused by mixing
of hydrothermal fluids with cold oxygenated seawater (Maslennikov et al. 2009). Host rocks also influence on high-tech trace element assemblages in chalcopyrite:
ultramafic (high Se, Sn, Co, Ni, Ag and Au) ! mafic (high Co, Se, Mo and low Bi, Au
and Pb) ! bimodal mafic (high Te, Au, Ag, Bi, Pb, Co, moderate Se, and variable As
and Sb) ! bimodal felsic (high As, Sb, Mo, Pb, moderate Bi, and low Co, Te and Se). In sphalerite of the same range, the contents of Bi, Ga, Pb, In, Ag, Au and Sb increase
versus Fe, Se, Sn and Co. The variations in pyrite coincide with these changes
(Maslennikov et al. 2017). 109 High-Tech Elements in Minerals of Massive Sulfide Deposits 4
Conclusions The mineralogical and trace element variations between chimneys are likely due to
increasing of O2 and decreasing temperature caused by mixing of hydrothermal fluids
with cold oxygenated seawater. The next stage of trace element differentiation is
halmyrolysis of sulfide sediments followed by diagenesis, anadiagenesis and meta-
morphism. Halmyrolysis as a leaching process of clastic sulfide sediments gives a way
to redeposition of high-tech element such as Se, Te, Au, Ag, Sn, and In as own
minerals in enrichment submarine supergene zones. The enrichment in the core of
sulfide nodules is changed to trace element depletion in later epigenetic stages. Local LA-ICP-MS analyses of the sulphides provides new insight into the pro-
cesses of selective ore treatment and recovery of high-tech elements from different
genetic ore varieties in implication to diverse ore formational types of massive sulfide
deposits. Aknowledgements. LA-ICP-MS analyses were carried out during visiting programs (2005,
2009, 2012, 2013, 2015) sponsored by the ARC Centre of Excellence grant to CODES. The
research of sulphide nodules was supported by the Russian Foundation for Basic Research
(project no. 17-05-00854) in the Institute of Mineralogy UB RAS. 110 V. Maslennikov et al. References Danyushevsky LV, Robinson R, Gilbert S, Norman M, Large R, McGoldrick P, Shelley JMG
(2011) Routine quantitative multi-element analysis of sulfide minerals by laser ablation ICP-
MS: standard development and consideration of matrix effects. Geochim Explor Environ Anal
11:51–60 Large RR, Danyushevsky L, Hillit H, Maslennikov V, Meffere S, Gilbert S, Bull S, Scott R,
Emsbo P, Thomas H, Singh B, Foster J (2009) Gold and trace element zonation in pyrite
using a laser imaging technique: implications for the timing of gold in orogenic and Carlin-
style sediment-hosted deposits. Econ Geol 104:635–668 Maslennikov VV, Maslennikova SP, Large RR, Danyushevsky LV (2009) Study of trace element
zonation in vent chimneys from the Silurian Yaman-Kasy VHMS (the Southern Urals,
Russia) using laser ablation inductively coupled plasma mass spectrometry (LA-ICP MS). Econ Geol 104:1111–1141 Maslennikov VV, Maslennikova SP, Ayupova NR, Zaykov VV, Tseluyko AS, Melekestseva IY,
Large RR, Danyushevsky LV, Herrington RJ, Lein AT, Tessalina SG (2017) Chimneys in
Paleozoic massive sulfide mounds of the Urals VMS deposits: mineral and trace element
comparison with modern black, grey, white and clear smokers. Ore Geol Rev 85:64–106 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction Native gold in exogenetic conditions, depending on its environment, undergoes gradual
changes, in morphology and material composition. It is known that, gold cleaning in
exogenetic conditions occurs mainly under chemical and physical-chemical influence
in weathering crusts, and also as a result of simple strain in geodynamic conditions. This report will be focused on gold cleaning as a result of impact of mechanogenic
processes in eolian conditions. 2
Methods and Approaches Chemical composition of eolian gold of the Lena-Viluy interfluve (east Siberian
platform) was studied by atomic-absorption spectrography (30 objects), spectral
quantitative analysis (50 objects), and at JXA-5OA micro-analyzer (30 objects and 200
identifications). Diamond and Precious Metal Geology Institute SB RAS, Yakutsk, Russia
znikiforova@yandex.ru Diamond and Precious Metal Geology Institute SB RAS, Yakutsk, Russia
znikiforova@yandex.ru Abstract. It is identified for the first time that, during process of complicated
deformation in eolian conditions - mechanical transformation of flaky gold into
toroidal form and then into globular-hollow form, gold is cleaned up to abso-
lutely pure metal with fineness 1000‰. Note that, fineness of this eolian gold is
higher than fineness of the reference object, shown by the detecting device
(JXA-5OA micro-analyzer). In this connection, identified natural process of
gold cleaning in eolian conditions can be successfully used in gold metallurgy to
obtain absolutely pure gold. Keywords: Gold Fineness Trace elements Structures Eolian process © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 111–114, 2019.
https://doi.org/10.1007/978-3-030-22974-0_25 Absolutely Pure Gold with High
Fineness 1000‰ Z. Nikiforova(&) Z. Nikiforova(&) Diamond and Precious Metal Geology Institute SB RAS, Yakutsk, Russia
znikiforova@yandex.ru 3
Results and Discussion During mechanical impact of sand grains on gold in eolian conditions, not only the
form is transformed, but also inner structure and chemical composition are regularly
changed. It is identified that, process of gold cleaning is more intensive in eolian
conditions, than in hydrodynamic conditions (Nikiforova 1999). 112 Z. Nikiforova Z. Nikiforova According to the analysis of eolian gold by atomic-absorption method, increase of
gold fineness during transformation of flaky forms into globular-hollow forms from 810
to 970‰ is identified. Flaky gold with scarcely noticeable elevation at the periphery,
has fineness range 810–970‰, with average gold fineness 890‰. In toroidal gold,
interval of gold fineness fluctuation is identified - 920–970‰, with average index
940‰. Globular-hollow form is characterized by high gold fineness - 960–990‰, with
average fineness 970‰. Spectral quantitative analysis identified that, constant trace elements of flaky forms
with characteristic features of eolian transformation are Fe- 0,1; Pb- 0,003; Sb- 0,002
Cu-0,017; Mn- 0,01; Pd- 0,002.; Ni-traces; Hg- traces. And some other. In toroidal gold,
a smaller set of trace elements is identified Fe- 0,1; Cu-0,02; Mn- 0,03; Ni- traces; Hg-
traces, and in globular-hollow body only these trace elements are identified – Fe- 0,1;
Cu-0,05; Mn- 0,001. Study of fineness of different areas of gold particles (Table 1 and Fig. 1) allowed
identifying that, a flake with medium-grained structure (grain C-9a) has fineness from
747 t0 780‰, and its more high-standard shell – from 950 to 988‰. Flake P-138 with
partially recrystallized rim has gold fineness 900–970‰, and its central part 814–860‰. Fully recrystallized flake showed maximum gold fineness – 990–1000‰. Fineness of
globular-hollow gold within one sample is not just high, but has an absolute value -
1000‰. For example, 17 identifications in the grain 60a found insignificant fluctuations
of fineness within interval 992–1000‰, and 13 identifications in the grain 60b showed
the highest gold fineness – 1000‰. Fig. 1. Section of gold particles and points of fineness identification Fig. 1. Section of gold particles and points of fineness identification Absolutely Pure Gold with High Fineness 1000‰ 113 Table 1. Z. Nikiforova Fineness of particular areas of gold particles, ‰
Points of
fineness
measurement
Morphologic type of eolian gold
Flaky
Globular recrystallized
Index of the
sample
C-9a
P-138
8642
Б-19a
B-19б
P-141a
P-141б
P-123
P-60a
P-60б
1
759
973
998
903
860
990
997
1000
996
1000
2
747
941
996
912
867
Not
identified
992
955
996
1000
3
759
933
998
907
869
983
997
999
1000
4
756
Not identified
984
904
919
990
991
999
1000
1000
5
757
908
994
912
988
985
996
999
1000
6
755
920
997
902
996
996
995
997
1000
7
758
860
996
901
980
987
1000
992
1000
8
769
892
997
999
984
938
992
1000
9
766
978
1000
980
970
930
999
1000
10
953
814
989
995
993
994
1000
11
783
896
994
985
997
1000
1000
12
979
806
993
995
995
1000
1000
13
986
956
990
987
not identified. 1000
1000
14
970
937
920
995
1000
15
974
973
932
1000
903
1000
16
988
974
994
988
912
1000
17
970
900
998
987
910
1000
18
987
1000
1000
1000
19
998
999
20
984
Not identified
21
991
22
1000
23
983
24
996
1000
25
998
1000
26
999
1000
27
1000
28
1000 Table 1. Fineness of particular areas of gold particles, ‰ It should be emphasized that, shell of globular-hollow forms constantly shows
absolutely high fineness 1000‰. Just in several samples, primary gold preserved in
partition, is characterized by lower fineness within a range from 860 to 919‰, for
example grain 19b. Increase of gold fineness in eolian conditions is explained by the fact that, as a
result of complex deformation of flaky gold, very thin films of gold (fraction of mcm)
are formed, being overlapped on each other, generate a shell of globular forms. In
addition, surface for active chemical interaction of metal with environmental elements
is increased, that contributed to the maximum removal of silver and trace elements
from primary gold particles. It should be emphasized that, shell of globular-hollow forms constantly shows
absolutely high fineness 1000‰. Just in several samples, primary gold preserved in 114 Z. Nikiforova partition, is characterized by lower fineness within a range from 860 to 919‰, for
example grain 19b. 4
Conclusions It is identified for the first time, that during complex deformation, when flaky gold is
mechanically transformed into toroidal form and then into globular-hollow form, metal
is cleaned to absolutely pure gold with fineness 1000‰. Identified natural process of
gold cleaning in eolian conditionsв can be successfully used in gold metallurgy to
obtain absolutely pure gold. Acknowledgements. The work is implemented within scientific-research projects of Diamond
and Precious Metal Geology Institute, Russian Academy of Sciences, project №0381-2016-
0004. Z. Nikiforova Increase of gold fineness in eolian conditions is explained by the fact that, as a
result of complex deformation of flaky gold, very thin films of gold (fraction of mcm)
are formed, being overlapped on each other, generate a shell of globular forms. In
addition, surface for active chemical interaction of metal with environmental elements
is increased, that contributed to the maximum removal of silver and trace elements
from primary gold particles. The process of mechanically gold cleaning was proved by (Lechtman 1979)
experimentally. According to the results of his experiments, clean layer of gold
appeared after multiple alteration of forging of gold and copper alloy, with its pro-
cessing in weak ammonia solution, where initial gold content did not exceed 12%. 1
Introduction Microhardness of gold was earlier studied by Lebedeva (1963), Badalova et al. (1968),
Petrovskaya (1968), Popenko (1982). Previous researchers identified that, microhard-
ness changes on the average from 40 to 100 kg/mm2 and depends on chemical com-
position of gold (percentage of silver and trace elements) and its inner structure. It is
known that, Ag and Cu trace elements significantly increase hardness of gold alloys. Trace element presence (Pt, Sn, Al) also causes sharp increase of gold hardness. Gold
of low fineness (550–650‰) has the biggest microhardness (Popenko 1982). References Lechtman H (1979) A pre-Columbian technique for electrochemical replacement plating of gold
and silver on copper objects. J Metals 31(12):154–160 Lechtman H (1979) A pre-Columbian technique for electrochemical replacement plating of gold
and silver on copper objects. J Metals 31(12):154–160
Nikiforova ZS (1999) Typomorphic features of eolian gold. ZVMO. N5, pp 79–83 Nikiforova ZS (1999) Typomorphic features of eolian gold. ZVMO. N5, pp 79–83 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Diamond and Precious Metal Geology Institute SB RAS, Yakutsk, Russia
znikiforova@yandex.ru Diamond and Precious Metal Geology Institute SB RAS, Yakutsk, Russia
znikiforova@yandex.ru Abstract. Microhardness of eolian gold – new morphologic type of placer
gold, is studied. Flaky gold particles with a elevation at the periphery, as well as
gold of toroidal and globular-hollow forms belong to eolian gold. Genesis of
eolian gold is related to mechanical transformation of flaky gold particles into
toroidal, and then into globular-hollow form in eolian conditions, that is
experimentally proven. Previous studies determined changes of microhardness,
mainly from 47 kg/mm2 to 100 kg/mm2, lower limit – 40 kg/mm2. But, low
microhardness was identified in globular-hollow gold for the first time, which
stood at 21 kg/mm2. This is due to the fact that, as a result of transformation of
flaky gold in eolian conditions, under mechanical and chemical processes, silver
and trace elements were removed, that led to fineness increase up to 1000‰, and
to decompaction of inner structure of gold, that influenced microhardness
indices. Identified patterns in nature, microhardness changes under mechano-
genic and chemical process impact in eolian conditions, can be successfully
used in gold metallurgy to produce gold alloys with very low microhardness. Keywords: Gold Microhardness Fineness Trace elements Morphology
Process Keywords: Gold Microhardness Fineness Trace elements Morphology
Process © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 115–118, 2019.
https://doi.org/10.1007/978-3-030-22974-0_26 New Data on Microhardness of Placer Gold Z. Nikiforova(&) 2
Methods and Approaches Microhardness of globular-hollow eolian gold was studied by microhardness tester
PMT-3. In order to detect microhardness, globular-hollow forms of gold were mounted
in epoxy specimen. In such a manner, optimum section of gold particles was obtained,
necessary for further study; it is possible to identify microhardness indices in its central
part, end and shell. 55 identifications were performed at 12 sections of globular-hollow
gold particles (Fig. 1 and Table 1). Since, gold fineness is increased in eolian Z. Nikiforova 116 conditions, high-standard shell is formed in end parts, and central parts remain
unchanged; microhardness was measured in end parts of the globular-hollow gold, and
in partition of hollow ball, being a relic of the flake. 3
Results and Discussion Microhardness of native gold, being in eolian conditions, is studied for the first time. It
is identified that, in four gold particles (1, 2, 3, 4), increase of microhardness from
partition center to the end is clearly observed. Microhardness between individual grains
of gold particles (1, 2) is 2 times different. Uniform microhardness is typical for
different parts of the globular-hollow forms (6, 7, 9). Sample 12 yields increase of
microhardness from outer part of the end to partition center. Other four gold particles
do not have any clear regularity concerning microhardness distribution. When microhardness of eolian gold was studied, it was found that, microhardness
depends on degree of inner structure changes and chemical composition of gold in
exogenetic conditions. When flaky gold is transformed in a complicated way into
toroidal form, and then into globular-hollow form in eolian conditions, not only gold
morphology is changed, but also chemical composition is changed, in particular gold
fineness is increased and amount of trace elements is decreased (Nikiforova 1999). It is
explained by the fact that, as a result of complicated transformation of flaky gold in
eolian conditions, very thin gold films are stretched from the flake end and overlap each
other, generating globular-hollow form. In this connection, surface for chemical impact
is increased in exogenetic conditions, that contributed to gradual removal of silver and
trace elements, and sharp decrease of microhardness indices of eolian gold. In addition, earlier unknown lower limit of microhardness of the globular-hollow
gold is identified - 21 kg/mm2. Fig. 1. Location of the points of identification of globular-hollow gold microhardness Fig. 1. Location of the points of identification of globular-hollow gold microhardness New Data on Microhardness of Placer Gold 117 Table 1. 3
Results and Discussion Microhardness of the elements of globular gold section, kg/mm2
Sample №
Elements of section of globular form
End 1
Partition
End 2
Base of the
Shell
End 1
End 2
I
64;5
4
41;2
2;3
55
1
II
25;7
3;4
21
2
28;6
1
III
41;2
6
25;7
5;4
32;1
1
55
7
41;2
2
25;7
3
23;2
8
IV
46;2
1
42;0
2;3
64;2
6
57;2
5
46;7
4
V
47;3
4
32;1
3
25;7
2
25;7
1
55
5
23;2
6
VI
43;0
1
41;2
2
VII
64;5
4
64;5
5
64;5
2
64;5
3
41;2
1
VIII
47;3
1
47;3
4
32;1
5
32;1
2;3
IX
28;6
1
28;6
28;6
3
X
47;3
1
55;0
2
XI
36;2
1
47;3
2
36;2
3
36;2
4
41;2
5
XII
47;3
3
55
2
41;2
1
36;2
4
32;1
5
*Above line – microhardness, below line – point of microhardness
measurement (Fig. 1). In general, it is true that microhardness of eolian gold relatively low in comparison
with native gold from other exogenetic conditions. It may be that, low microhardness is
due to decompaction of some parts of the shell and the end of globular-hollow gold. Petrovskaya (1973) identified that, structures of recrystallization cause decompaction,
stress relief, removal of silver and trace elements, that lead to increase of gold fineness
up to 1000‰. Structures of decompaction, high fineness and paucity of trace elements
are observed in the studied gold, that is why microhardness indices of globular-hollow
gold reached such low limits. In addition, earlier unknown lower limit of microhardness of the globular-hollow
gold is identified - 21 kg/mm2. In general, it is true that microhardness of eolian gold relatively low in comparison
with native gold from other exogenetic conditions. It may be that, low microhardness is
due to decompaction of some parts of the shell and the end of globular-hollow gold. Petrovskaya (1973) identified that, structures of recrystallization cause decompaction,
stress relief, removal of silver and trace elements, that lead to increase of gold fineness
up to 1000‰. Structures of decompaction, high fineness and paucity of trace elements
are observed in the studied gold, that is why microhardness indices of globular-hollow
gold reached such low limits. 118 Z. Nikiforova 4
Conclusions Thus, low microhardness – 21 kg/mm2 is identified in globular-hollow gold is iden-
tified for the first time. Identified patterns in nature, microhardness changes under
mechanogenic and chemical process impact in eolian conditions, can be successfully
used in gold metallurgy to produce gold alloys with very low microhardness. Acknowledgements. The work is implemented within scientific-research projects of Diamond
and Precious Metal Geology Institute, Russian Academy of Sciences, project №0381-2016-0004. References Badalova RP, Nikolaeva EP, Tolkacheva LF (1968) Study of microhardness of minerals of the
silver-gold series from gold deposits of Uzbekistan. “Physical features of rare-metal minerals
and methods of their study” collected articles, Moscow, Nauka Filippov VE, Nikiforova ZS (1988) Transformation of native gold particles during process of
eolian impact. AN SSSR Report, vol 299, №5, pp 1229–1232 Lebedeva SI (1963) Identification of microhardness of minerals. AN SSSR Publishing House,
Moscow Nikiforova ZS (1999) Typomorphic features of eolian gold. ZVMO, N5, pp 79–83 Petrovskaya NV (1973) Native gold. Nedra, Moscow Popenko GS (1982) Mineralogy of fold from the Quaternary deposits of Uzbekistan, Tashkent,
FAN Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. E. Ozhogina(&), A. Rogozhin, O. Yakushina, Yu. Astakhova,
E. Likhnikevich, N. Sycheva, A. Iospa, and V. Zhukova FSBE “All-Russian Institute of Mineral Raw Materials” (VIMS),
Moscow, Russia
vims@df.ru FSBE “All-Russian Institute of Mineral Raw Materials” (VIMS),
Moscow, Russia
vims@df.ru Abstract. Modern technologies of mineralogical study and evaluation of rare
metal raw quality are focused on its variety. Methods of the mineral processing,
allowing to optimize monitoring of ore properties defining technological pro-
cesses and quality of expected products are presented. Some examples of rare
metal ores mineralogical study are given. The main challenging tasks in rare
metal ores quality evaluation are considered. Keywords: Technological mineralogy Methods of analysis Testing
Rare metals Minerals Ores Specific features Quality assurance
Quality assessment Keywords: Technological mineralogy Methods of analysis Testing
Rare metals Minerals Ores Specific features Quality assurance
Quality assessment Keywords: Technological mineralogy Methods of analysis Testing © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 119–122, 2019.
https://doi.org/10.1007/978-3-030-22974-0_27 Modern Methods of Technological Mineralogy
in Assessing the Quality of Rare Metal
Raw Materials E. Ozhogina(&), A. Rogozhin, O. Yakushina, Yu. Astakhova,
E. Likhnikevich, N. Sycheva, A. Iospa, and V. Zhukova 2
Methods and Approaches The variety and complexity of rare metal ores mineralogical features identify the
necessity to use a set of modern physical research methods of analysis to get reliable
data on their composition and structure. This complex of mineralogical methods is
individually selected depending on ores features and research tasks. Mineralogical study of rare metal ores is necessary at all stages of deposits exploration
and development. We particularly note the importance of ore mineralogical and analytical
study of at the early stages of geological exploration, that allow to carry out technological
assessment of raw materials with minimal investment, and at deposit exploration,
involving geological and technological mapping for the detailed study of the mineral-
ization zoning, minerals and mineral associations distribution, variations in the ore-
bearing phases properties and characteristics, identification of the technological types and
species whiting the geological margins of the deposit. The research practice proves great
contribution of mineralogical study in deposits investigation and quality assessment. When mineralogical investigation is the result of a set of implemented methods,
including not only usually traditional ones (optical microscopy, radiography), but also
precise analysis (analytical electron microscopy, microprobe). Mineral and techno-
logical mapping challenge today the Zashikhinsky, Tomtor, Chuktukon and other
deposits of rare earth elements. 1
Introduction Variety of rare metal raw materials is determined, firstly, by a significant amount of
industrial minerals, among which there are more than 20 main and about 30 minor and
accessory minerals of different genesis; secondly, by the diversity of their genesis:
magmatic, pegmatite, greisen, scarn, metamorphic, hydrothermal, sedimentary, hyper-
geneous. The rare metal-bearing minerals are the ones that contain Tantalum, Niobium,
Bismuth, Tellurium, Zirconium, Hafnium, Yttrium, Scandium, Lanthanides, Lithium,
Beryllium, Cesium, Rubidium, Strontium, Barium. A lot of rare metal ores, mainly
tantalum-niobium, rare-earth phosphate, carbonate and silicate, and frequently zirco-
nium, are radioactive. The vast majority of rare-metal ores are complex, and industrial
minerals can be both main, and secondary, subordinate ones. General features of the rare
metal-bearing ores are as follows: – complex texture-structural pattern (a significant number of fine, metacolloidal spots
formed by minerals and aggregates of micro-and nanometer size); – complex texture-structural pattern (a significant number of fine, metacolloidal spots
formed by minerals and aggregates of micro-and nanometer size); – polymineral composition associated with the simultaneous presence of minerals
developed in different paragenetic associations; – polymineral composition associated with the simultaneous presence of minerals
developed in different paragenetic associations; – variations in chemical composition of the ore-forming minerals caused by chemical
elements isomorphic substitutions in their crystal structure; – variations in chemical composition of the ore-forming minerals caused by chemical
elements isomorphic substitutions in their crystal structure; © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 119–122, 2019. https://doi.org/10.1007/978-3-030-22974-0_27 © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 119–122, 2019. https://doi.org/10.1007/978-3-030-22974-0_27 E. Ozhogina et al. 120 – mineral grains phase heterogeneity of various origin, namely decay of solid solu-
tions, syngeneic inclusions, zonal growth, multiple stages of generation, partial
recrystallization, secondary solid-phase transformations, etc.; – mineral grains phase heterogeneity of various origin, namely decay of solid solu-
tions, syngeneic inclusions, zonal growth, multiple stages of generation, partial
recrystallization, secondary solid-phase transformations, etc.; – ore minerals with radioactive elements can undergo transformations resulting in
metamictic forms origin (disrupted crystal structure due to radiation damage) or
partially metamictic (damaged crystal structure due to radiation). – ore minerals with radioactive elements can undergo transformations resulting in
metamictic forms origin (disrupted crystal structure due to radiation damage) or
partially metamictic (damaged crystal structure due to radiation). 3
Results and Discussion The characteristic feature of technological mineralogy is integration/conjunction of
research methods and modern technical means/units. It is especially important when
studding the rare-metal mineralization, because it is not always possible to uniquely
identify industrially valuable minerals, to establish the mineral form of useful compo-
nents, to identify and study characteristics of minerals appearing in the fine aggregates. Pyrochlore-Mmonazite-Crandallite ores of the Tomtor deposit differ in specific
composition, are rich in content and reserves of REE, Niobium, Yttrium, Scandi- um,
Phosphorus and are a non-standard type of rare-metal raw materials. Ores features are
the variable granular composition, often high dispersion, polymineral composition,
various forms of occurrence the rare earths and niobium-bearing minerals, the character
of their localization in close association with Alumiium- Silicate minerals. The ores are
formed by polymineral aggregates with variable content of Crandallite and Kaolinite of
fine-grained structure. The aggregate cement contains also Siderite, Ilmenorutil, Modern Methods of Technological Mineralogy 121 Anatase, Pyrochlore, iron hydroxides, and other minerals. Most often the aggregates
dispose earthy-type structure. Pyrochlore is the main Niobium mineral, that form both individual octahedral
crystals, fragments of rounded and angular forms, as well as aggregates of tiny grains in
size ranging from 1 lm to 0.5 mm, in varying degrees transformed by hypergene
processes. The rock is distributed in the form of microinclusions, forming a “rash” in
the cementing mass of aluminophosphates and silicates. According to microprobe data
(X-ray microspectral microanalysis), the main feature of the hypergenic alteration of
Pyrochlore is the replacement of Ca and Na by Sr, Ba and Pb; the altered Pyrochlore
varieties significantly dominate in the ores. Hypergeneous transformation of Pyrochlore
was accompanied by textural transformations, typically clearly manifested in the dis-
integration of its large crystals into small blocks (Fig. 1). Often the cracks between the
separate Pyrochlore individuals are filled by Crandallite group minerals and Apatite,
rarely by sulfides. Fig. 1. Disintegrated crystals of Pyrochlore. TEM, image in back scattered electrons Fig. 1. Disintegrated crystals of Pyrochlore. TEM, image in back scattered electrons Minerals of Crandallite group mostly form polymineral aggregates being dominant
phase in these aggregates. There is a weak individualization of isometric and round
shape grains, closely associated with fine Pyrochlore, Monazite, iron hydroxides, their
grain size in the ores is often much less than 5 micrometers. According to microprobe
analysis, the Crandallite group minerals have a mixed composition. 3
Results and Discussion According to X-ray
powder diffraction analysis (XPD), the interplanar distances d(hkl) reliably identificate
Gorceixite (2.96, 3.55, 5.72 Å) and the intermediate Goyazite-Florencite series (2, 94,
3, 53, of 5.71 Å). Mineralogical features of a Pyrochlore-Monazite-Crandallite ores (variable granular
composition, often with high dispersion; polymineral composition due to the simul-
taneous presence of different paragenesis assemblages, different form of Niobium and
rare earths presence, vide range of isomorphic substitutions in the structure of minerals,
the proximity of their physical properties) determine the impossibility of these ores
processing by methods of deep enrichment. Therefore, the prospects of such ores
processing should be associated with hydrometallurgy. Typical features of the Chuktucon Deposit rare earth ores were established on
electron microscopic study. The main ore minerals are Pyrochlore, Monazite and the 122 E. Ozhogina et al. Crandallite group minerals. All of them are superfinely dispersed and are in close
assemblage/association with Iron and Manganese oxides and hydroxides. The latter
form complex types of accretions with Pyrochlore (corrosive), Monazite (envelope of
Goethite around Monazite grains), Crandallite group minerals (thin jointing), which
negatively affects their disclosure and does not allow to identify and study these
minerals by traditional methods of optic microscopy. Microprobe study indicated
varieties of secondary Pyrochlore (bearing Cerium, Barium, Strontium and mixed
type), and variable chemical composition of the Crandallite group minerals. Almost
constant presence of Iron and Manganese mechanical impurities was established in all
ore minerals. An independent mineral form of Cerium – Cerianite has been identificate
by X-ray powder diffraction data. 4
Conclusions The main challenging tasks for the rare metal ores investigation and quality evaluation
should be considered: • predictive mineral and technological assessment of raw materials of natural and
man-made origin; • predictive mineral and technological assessment of raw materials of natural and
man-made origin; • geological and technological mapping using an Arsenal of methods of technological
Mineralogy; • forecasting of technological properties of ores at various stages of processing and
development of methods of their directed change; • increase of complexity of development of deposits and deep processing of ores;
• identification and involvement in the industrial use of non-traditional types of rare
metal raw materials; ;
assessment of environmental consequences of industrial development of deposits. assessment of environmental consequences of industrial development of deposits. Therefore, the main task of technological mineralogy in the study of the rare metal
ores is today their comprehensive study for quality assessment at all stages of geo-
logical research and development of mineral deposits. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Topochemical Transformations
in Sodium-Bismuth-Silicate System
at 100–900 °C A. Pavlenko(&) and R. Yastrebinskiy Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
yrndo@mail.ru Abstract. The authors have developed a method for producing highly dis-
persed sillenite bismuth silicate in the system Na2O-Bi2O3-SiO2 (NBS) from
water solutions of organosilicon monomers (sodium methylsiliconate) and bis-
muth nitrate. The paper studies the phase composition and microstructure of the
synthesized NBS material at different temperatures. The morphology of crystals
in the NBS material and the peculiarities of its thermal-oxidative breakdown are
investigated. X-ray diffraction spectra obtained using a cuКa-source are used to
evaluate the crystal lattice spacing and to analyze the broadening of the
maximum-intensity diffraction line for this crystal with due consideration of
crystal indices h, k, l by the approximation method to determine the dimensions
of the coherent scattering region and microdistortions of the crystal lattice Da/a. The authors established that the silicate shell on Bi12SiO20 particles is close to
the silicates with continuous chain radicals [SiO3]∞
2−, and a part of them are
bridges between the bismuth silicate particles. Keywords: Sillenite Phase composition Microstructure
Crystal morphology Thermal treatment Crystal lattice microdeformation Keywords: Sillenite Phase composition Microstructure
Crystal morphology Thermal treatment Crystal lattice microdeformation © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 123–126, 2019.
https://doi.org/10.1007/978-3-030-22974-0_28 2
Materials and Methods The authors have developed a method for producing highly dispersed sillenite bismuth
silicate in system Na2O-Bi2O3-SiO2 from water solutions of organosilicon monomers
(sodium methylsiliconate) and bismuth nitrate (Yastrebinskii et al. 2018). The amounts of the components were calculated with the aim of producing stable
bismuth silicate Bi12SiO20 (6Bi2O3 SiO2). At 100 °C we have obtained highly dispersed (0.2–0.3 lm) hydrophobic NBS
material that is insoluble in water (NBS wetting angle is 122°). The density is 3780 kg/m3. At 100 °C we have obtained highly dispersed (0.2–0.3 lm) hydrophobic NBS
material that is insoluble in water (NBS wetting angle is 122°). The density is 3780 kg/m3. According to mass-spectroscopy, NBS material had the following composition
(expressed as oxides), wt%: Na2O - 23.83; Bi2O3 - 59.70; SiO2 - 16.47. According to mass-spectroscopy, NBS material had the following composition
(expressed as oxides), wt%: Na2O - 23.83; Bi2O3 - 59.70; SiO2 - 16.47. Differential thermal analysis (DTA) and thermogravimetric analysis (TGA) of
specimens were performed on a STA-449 F1 Jupiter derivatograph (Germany). X-ray
diffraction (XRD) analysis of phases and structure was performed on a ARL™X’TRA
Powder Diffractometer (Switzerland) with Cukx source (kkx = 1.542 Å) using a nickel
filter. The infrared spectra were obtained on a Specord-75IR spectrometer (Germany). The material microstructure was studied by raster electron microscopy (REM) in the
modes of reflected (back-scattered) and secondary electrons. 1
Introduction The development of highly dispersed metal-organosiloxane fillers with modified sur-
face allows solving a multitude of important problems in the field of radiation materials
science (Pasechnik 2006). The promising approach is to use water-soluble chemically
active organosiloxanes as the basis for production of metal oligomers. A new tech-
nological approach to the solution of the stated complex problem is required. As of today, the chemistry of organosiloxane compounds of bismuth attract par-
ticular attention. This is conditioned by multiple valuable properties of organosilicon
compounds (high thermal stability, hydrophobicity, dielectric characteristics and
resistance to a range of aggressive media). Besides, bismuth atoms have large capture
cross-section of gamma-radiation, which is almost the same as for lead atoms in a wide
energy spectrum. The presence of vacant 3d-orbitals in silicon atoms conditions high
reactivity of bond Si-OH in silicate minerals. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 123–126, 2019. https://doi.org/10.1007/978-3-030-22974-0_28 124 A. Pavlenko and R. Yastrebinskiy 3
Results and Discussion Phase Composition and Microstructure of Mineral Phases in NBS Material
Synthesized at 100 °C. XRD phase analysis using the Powder Diffraction File
(PDF) and literature data (Gorshkov and Timashev 1981; Gorelik et al. 2002) allowed
detecting the formation of three amorphous-crystalline mineral phases: 1. Metastable bismuth silicate Bi2SiO5 (d = 3.0379 Å/I = 100%; 3.7169 Å; 2.7223 Å)
with tetragonal crystal system (a = 3.802; c = 15.134 Å), with the amorphous ring
of about 3 Å. 1. Metastable bismuth silicate Bi2SiO5 (d = 3.0379 Å/I = 100%; 3.7169 Å; 2.7223 Å)
with tetragonal crystal system (a = 3.802; c = 15.134 Å), with the amorphous ring
of about 3 Å. 2. Bismuth oxide a-Bi2O3 (d = 3.2596 Å/I = 100%; 3.2596 Å; 1.9625 Å) with
monoclinic crystal system (a = 5.8499; b = 8.1698; c = 7.5123 Å) with the
amorphous ring of about 3 Å. 2. Bismuth oxide a-Bi2O3 (d = 3.2596 Å/I = 100%; 3.2596 Å; 1.9625 Å) with
monoclinic crystal system (a = 5.8499; b = 8.1698; c = 7.5123 Å) with the
amorphous ring of about 3 Å. 3. Bismuth organosilicate H3C(SixBiyOz)Na with the amorphous ring of 10–12 10–
12 Å and clear X-ray reflection at d = 11.4513 and 5.7090 Å. However, the precise
determination of this composition using PDF failed. 3. Bismuth organosilicate H3C(SixBiyOz)Na with the amorphous ring of 10–12 10–
12 Å and clear X-ray reflection at d = 11.4513 and 5.7090 Å. However, the precise
determination of this composition using PDF failed. Fig. 1. IR spectrum of synthesized NBS material Fig. 1. IR spectrum of synthesized NBS material 125 Topochemical Transformations in Sodium-Bismuth-Silicate System The results of IR-spectroscopy, the silicate phases in NBS powder synthesized at
100 °C have linear structure. The splitting of the absorption bands in the range of
1000–1100 cm−1 that is typical for the siloxane bond indicates the presence of several
types of siloxane phases (Fig. 1). Morphology of Crystals in Synthesized NBS Material. According to REM, NBS
material synthesized at 100 °C contained particle agglomerations of irregular shape
with the size of 0.8–2.5 lm (Fig. 2). Fig. 2. Electron microphotographs (REM) of NBS material synthesized at 100 °C Fig. 2. Electron microphotographs (REM) of NBS material synthesized at 100 °C Defectiveness of Crystals in NBS Material Subjected to Thermal Treatment. Defectiveness of Crystals in NBS Material Subjected to Thermal Treatment. X-ray diffraction spectra obtained using a CuКa-source were used to evaluate the
crystal lattice spacing and to analyze the broadening of the maximum-intensity
diffraction line for this crystal with due consideration of crystal indices h, k, l by the
approximation method to determine the dimensions of the coherent scattering region
and microdistortions of the crystal lattice Da/a. At 100–300 °C XRD analysis detected amorphous-crystalline bismuth organosili-
conate H3C(SixBiyOz)Na with the amorphous ring of 10–12 Å and clear X-ray
reflection at d = 11.4810 Å and 5.7020 Å. At the temperature of 200 °C, bismuth silicate Bi2SiO5 in the mixture of minerals
in terms of X-ray parameters approaches to the benchmark silicate of this composition
(card no. 36-288 PDF: d = 3.0400 Å (I = 100%, hkl = 103). In
the
temperature
interval
of
300–400 °C,
metastable
bismuth
silicate
Bi12Si0,87O20 with cubic crystal lattice in the synthesized dry mix transforms into stable
bismuth silicate Bi12SiO20 also with cubic lattice. In the temperature interval of 300–500 °C, the density of dislocations in the
structure of bismuth silicate Bi12SiO20 crystals was lowering, while at the temperature
higher than 650 °C, it was conversely rising up. The increase of the temperature from
300 to 500 °C improves the parameters of elementary crystal lattice of bismuth silicate
Bi12SiO20 by 0.0357 Å and 0.0268 Å, as compared to the benchmark crystal. The
volume of elementary crystal cell in this temperature interval grows by 2% and
amounts to 1040.5870 Å3. 126 A. Pavlenko and R. Yastrebinskiy A. Pavlenko and R. Yastrebinskiy 4
Conclusion In the study, a method for producing highly dispersed sillenite bismuth silicate in the
system Na2O-Bi2O3-SiO2 (NBS) from water solutions of organosilicon monomers
(sodium methylsiliconate) and bismuth nitrate was developed. The paper studied the
phase composition and microstructure of the synthesized NBS material at different
temperatures. The crystalline structure of the substance and the presence of silicate
amorphous phase in it were discovered. The paper revealed the morphology of crystals
in the synthesized NBS material and the peculiarities of its thermal-oxidative break-
down; the silicate shell on the particles of Bi12SiO20 was close to continuous chain
radicals SiO3
½
2
1 and a part of them played the role of bridges between bismuth silicate
particles. The determination of physicochemical properties of modified Bi12SiO20 sil-
lenite crystals was of appreciable significance. Acknowledgements. The work is realized in the framework of the Program of flagship uni-
versity development on the base of Belgorod State Technological University named after V.G. Shukhov, using equipment of High Technology Center at BSTU named after V.G. Shukhov, the
project within strategic development program No. a-51/17. 1
Introduction The Nevenrekan Au-Ag deposit is located in the Severo-Even district of the Magadan
region, Russia. The site is located within the central part of the Okhotsk-Chukotka
volcanogenic belt. The main structural element is cretaceous-paleogenic Nevenrekan
intrusive dome, which is crossed by quartz-adularia and quartz-carbonate veins with
hydrothermal origin. These veins contain ores with Au-Ag mineralization. This work is
motivated by mineralogical criteria for understanding ore-forming processes which are
essential for future efficient exploration. Ag-Bearing Mineralization of Nevenrekan
Deposit (Magadan Region, Russia) 1 “RMRL” Ltd. (Raw Materials Researching Laboratory),
Saint Petersburg, Russia
podolyan@lims-lab.com
2 Department of Mineral Deposits, Saint Petersburg State University,
Saint Petersburg, Russia 1 “RMRL” Ltd. (Raw Materials Researching Laboratory),
Saint Petersburg, Russia
podolyan@lims-lab.com
2 Department of Mineral Deposits, Saint Petersburg State University,
Saint Petersburg, Russia Abstract. Nevenrekan deposit, located in Magadan region, Russia, is a per-
spective Au-Ag deposit with average silver content 445 ppm and gold 7.4 ppm. The study of the ores reveals the main Ag-bearing minerals: Au-Ag alloys,
tetrahedrite-tennantite series and new phase – Ag8SnSe2S4. New phase contains
the main part of the silver of the deposit – 97.6%. Keywords: Silver Ag-sulphosalts Ag-bearing minerals Epithermal
Argyrodite References Gorelik SA, Skakov YuA, Rastorguev LN (2002) X-radiography and electron-optical analysis
(in Russian). MISIS, Moscow, 360 p Gorshkov VS, Timashev VV (1981) Methods of physicochemical analysis of silicates
(in Russian). Higher School, Moscow, 335 p Pasechnik OF (2006) Study of properties and structure of polyamide films after interaction of
low-orbit space factors. Obninsk, p 113. (in Russian) Yastrebinskii RN, Bondarenko GG, Pavlenko VI (2018) Synthesis of stable bismuth silicate with
sillenite structure in the Na2O–Bi2O3–SiO2 system. Inorg Mater Appl Res 9(2):221–226 Yastrebinskii RN, Bondarenko GG, Pavlenko VI (2018) Synthesis of stable bismuth silicate with
sillenite structure in the Na2O–Bi2O3–SiO2 system. Inorg Mater Appl Res 9(2):221–226 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 127–128, 2019.
https://doi.org/10.1007/978-3-030-22974-0_29 2
Methods and Approaches Methods of research include: (1) chemical analyses and its interpretation (include
X-Ray diffraction, atomic absorption analysis, inductively coupled plasma mass-
spectrometry (ICP-MS) analysis for micro components and X-Ray fluorescence anal-
ysis for macrocomponents); (2) optical researches including petrographic research of
thin sections and mineragraphic research of polish sections which are provided by
electron microprobe analyses of chemical composition of ore minerals; (3) technolog-
ical experiments (gravity и floatation methods of ores separation). © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 127–128, 2019. https://doi.org/10.1007/978-3-030-22974-0_29 128 E. Podolian et al. 4
Conclusions The main Ag-bearing minerals at the Nevenrekan deposit are Au-Ag alloys, tennantite-
tetrahedrite series and Ag8SnSe2S4 sulphosalt of argyrodite group, which contain
97.6% of silver. The mineral composition of ores and the mineral balance of silver
determine the prospects of flotation flow sheet for ore beneficiation. Acknowledgements. We thank Polymetal Engineering Company for the ore samples and RMRL
Ltd. for sponsorship of the participation in 14th International Congress for Applied Mineralogy. 3
Results and Discussion The main types of rocks at the deposit are clastolaval rhyodacites and ignimbrite rhy-
odacites, which are host rocks, and quartz-adularia veins, which are gangue rocks. Ore
mineralization of Nevenrekan deposit consists of 4 ore associations which are charac-
terized by different temperature of formation: 1. rutile-kassiterite; 2. pyrite-arsenopyrite;
3. sphalerite-chalcopyrite-galena-Au-Ag alloys-stannite series-Ag8SnSe2S4-tennantite-
tetrahedrite series with Ag; 4. hydrohematite. The most important paragenesis is the
third one because it includes the main mineral concentrators of silver and gold. Tech-
nological and following chemical and mineralogical researches of beneficiaries
demonstrate that the main concentrator of silver is sulphosalts, especially new phase
Ag8SnSe2S4, which contains 97.6% of all silver. According to crystallographic
researches, phase Ag8SnSe2S4 is orthorhombic and should belong to argyrodite group
(Zhai et al. 2018). Nevenrekan deposit belongs to low-sulfidation type in classification
of epithermal deposits (White and Hedenquist 1995). References Zhai D, Bindi L, Voudouris P, Liu J, Tombros S, Li K (2018) Discovery of Se-rich canfieldite,
Ag8Sn(S,Se)6, from the Shuangjianzishan Ag-Pb-Zn deposit, NE China: a multimethodic
chemical and structural study. Miner Mag 1–21 y
g
White NC, Hedenquist JW (1995) Epithermal gold deposits styles, characteristics and
exploration. SEG Newsl 23:9–13 White NC, Hedenquist JW (1995) Epithermal gold deposits styles, characteristics and
exploration. SEG Newsl 23:9–13 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction Precambrian formations, especially Pre-Riphean, which underwent metamorphism, as a
rule, experienced it repeatedly, i.e. were polymetamorphic. Accordingly, the zircons of
the newly formed, or transformed from the previously existing ones, in the course of
these events should have acquired some new properties, expressed in changes in the
morphology of the crystals, the internal structure, the geochemical composition. That is
what we see in zircons from various polymetamorphic complexes of the Urals,
including those located in its northern part: Nyartin, in the Subpolar Urals, and Harbey
in the Polar Urals, where up to five morphological types of this mineral are distin-
guished (Pystina and Pystin 2002). Th/U Relations as an Indicator of the Genesis
of Metamorphic Zircons (On the Example
of the North of the Urals) Y. Pystina(&) and A. Pystin
Institute of Geology Komi SC UB RAS, Syktyvkar, Russia
pystina@geo.komisc.ru Y. Pystina(&) and A. Pystin Abstract. Having studied polymetamorphic complexes of the Urals, including
its northern part, for many years we have collected material on the basis of
which we attempted to make generalizations concerning both the morphology of
zircons and their geochemical features, allowing the mineral to be used in the
reconstruction of specific metamorphic events and the interpretation of
geochronological data (Pystina et al. 2017; Pystina and Pystin 2002). In recent
years we also obtained new results on the morphology and geochemistry of
zircons from granitoids in the northern part of the Subpolar Urals (Pystina and
Pystin 2002). Together, this made it possible to compare different morphological
types of magmatic and metamorphic zircons. Keywords: Polymetamorphic complexes Geochronological Morphology
Geochemistry of zircons Keywords: Polymetamorphic complexes Geochronological Morphology
Geochemistry of zircons © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 129–132, 2019.
https://doi.org/10.1007/978-3-030-22974-0_30 2
Results and Discussion The same picture, according to our data, is observed in zircons from the rocks of
the metamorphic complexes of the Southern Urals: the Alexandrov and Ilmenogorsky
(Selyankin block). The content and distribution of the scattered elements according to
the data available today with morphological types of zircons are not clearly correlated. In this regard, the Th/U ratio deserves interest, which for magmatic zircons are, as a
rule, > 0.5 (Skublov et al. 2009), and for metamorphic, significantly lower – 0.1–0.3
(Pystin and Pystina 2015a), according to Rubatto < 0.07 (Rubatto 2002), although
according to other data it can be > 0.5, for example, 0.73 in zircons from the eclogite of
the Maksyutov Complex (Pystin and Pystina 2015b). But, despite some rebounds in
Th/U values, the average indices for metamorphic zircons, as well as for magmatic ones,
are fairly consistent. In the metamorphic zircons of the gneisses of the Nyartin Complex,
obtained from 9 crystals, they vary from 0.02 to 0.39, but among these values, two are
essentially out of the general picture – 0.75 and 0.68. Such values of the Th/U ratio
correspond to zircons of magmatic origin. In our case, these are zircons of prismatic
habitus, which we have isolated into the “migmatite” type. The isotope age, obtained by
the U-Pb SHRIMP-II method, is 503 ± 8 Ma and 498 ± 8 Ma, respectively (Pystin
and Pystina 2008). Their formation was associated with the metamorphism of the
amphibolite facies and the associated granitization. Therefore, the conditions, under
which this morphotype was formed, were similar to the conditions of granite formation,
hence we assume high Th/U ratios. An even clearer picture is obtained for zircons of the “granulitic” type from the
130
Y. Pystina and A. Pystin Y. Pystina and A. Pystin 130 conditions that do not exceed the low to medium stages of the amphibolite facies. In
more high-temperature conditions, it is found in the rocks of the basic series. The reason
for the emergence of such intricate forms of zircon may be the absence or deficiency of
silicate melt. Therefore, when P-T reaches the conditions of metamorphism sufficient for
the development of migmatization processes, such zircon can continue to crystallize
only in metamorphic mafic composition, for which, as is known, the migmatization
temperature is higher. Opaque zircons (type 5) are associated with the manifestation of
medium temperature diaphtoresis. 2
Results and Discussion The internal structure of all morphotypes of zircons is
characterized by the presence of nuclei of irregular or rounded shape, in the “migma-
titic” type, oscillatory zonality is usually noted, the “granulitic” type is the most
homogeneous. Among the isolated morphotypes of zircons in polymetamorphic com-
plexes, the “granulite” and “migmatite” types prevail. If, by morphological features and
internal structure, the zircons of polymetamorphic complexes are surely divided into
morphotypes, which can be associated with certain metamorphic events or processes,
then the geochemical composition of the scattered elements does not make a clear
separation. On the one hand, this is due to the extremely low content of the elements
themselves, most often zircon is enriched only in Hf, Y, U, P. C, on the other hand, the
nature of the distribution of these elements in the crystal, their quantitative variations do
not give grounds to separate some zircon morphotypes from others. Although in some
cases it is possible. E.g., in “migmatite” zircons (type 4) from the gneiss of the Harbay
Complex, the distribution of Hf decreases from the center of the crystal to the edge,
while in other morphotypes of zircons from the same rocks it increases. At the same
time, in the gneisses of the Nyartin Complex in all the selected morphotypes, including
the “migmatite” type, the content of Hf increases from the center of the crystal to the
edge. The same picture, according to our data, is observed in zircons from the rocks of
the metamorphic complexes of the Southern Urals: the Alexandrov and Ilmenogorsky
(Selyankin block). The content and distribution of the scattered elements according to
the data available today with morphological types of zircons are not clearly correlated. In this regard, the Th/U ratio deserves interest, which for magmatic zircons are, as a
rule, > 0.5 (Skublov et al. 2009), and for metamorphic, significantly lower – 0.1–0.3
(Pystin and Pystina 2015a), according to Rubatto < 0.07 (Rubatto 2002), although
according to other data it can be > 0.5, for example, 0.73 in zircons from the eclogite of
the Maksyutov Complex (Pystin and Pystina 2015b). But, despite some rebounds in
Th/U values, the average indices for metamorphic zircons, as well as for magmatic ones,
are fairly consistent. 2
Results and Discussion Detritic zircons (type 1) determine the metamorphic affiliation to one or another source
formation. Zircons of the “soccer ball” type or, as is customary in the Urals, after
Krasnobayev (1986), call them “granulitic” (type 2), and also “migmatite” (type 4) fix
several age levels of occurrences of high-temperature rock transformations. Zircon of
irregular shape like “cauliflower” (type 3) is typical for rocks metamorphosed under conditions that do not exceed the low to medium stages of the amphibolite facies. In
more high-temperature conditions, it is found in the rocks of the basic series. The reason
for the emergence of such intricate forms of zircon may be the absence or deficiency of
silicate melt. Therefore, when P-T reaches the conditions of metamorphism sufficient for
the development of migmatization processes, such zircon can continue to crystallize
only in metamorphic mafic composition, for which, as is known, the migmatization
temperature is higher. Opaque zircons (type 5) are associated with the manifestation of
medium temperature diaphtoresis. The internal structure of all morphotypes of zircons is
characterized by the presence of nuclei of irregular or rounded shape, in the “migma-
titic” type, oscillatory zonality is usually noted, the “granulitic” type is the most
homogeneous. Among the isolated morphotypes of zircons in polymetamorphic com-
plexes, the “granulite” and “migmatite” types prevail. If, by morphological features and
internal structure, the zircons of polymetamorphic complexes are surely divided into
morphotypes, which can be associated with certain metamorphic events or processes,
then the geochemical composition of the scattered elements does not make a clear
separation. On the one hand, this is due to the extremely low content of the elements
themselves, most often zircon is enriched only in Hf, Y, U, P. C, on the other hand, the
nature of the distribution of these elements in the crystal, their quantitative variations do
not give grounds to separate some zircon morphotypes from others. Although in some
cases it is possible. E.g., in “migmatite” zircons (type 4) from the gneiss of the Harbay
Complex, the distribution of Hf decreases from the center of the crystal to the edge,
while in other morphotypes of zircons from the same rocks it increases. At the same
time, in the gneisses of the Nyartin Complex in all the selected morphotypes, including
the “migmatite” type, the content of Hf increases from the center of the crystal to the
edge. 2
Results and Discussion In the metamorphic zircons of the gneisses of the Nyartin Complex,
obtained from 9 crystals, they vary from 0.02 to 0.39, but among these values, two are
essentially out of the general picture – 0.75 and 0.68. Such values of the Th/U ratio
correspond to zircons of magmatic origin. In our case, these are zircons of prismatic
habitus, which we have isolated into the “migmatite” type. The isotope age, obtained by
the U-Pb SHRIMP-II method, is 503 ± 8 Ma and 498 ± 8 Ma, respectively (Pystin
and Pystina 2008). Their formation was associated with the metamorphism of the An even clearer picture is obtained for zircons of the “granulitic” type from the
Alexandrov Complex in the Southern Urals, where the Th/U ratio varies from 0.23 to Th/U Relations as an Indicator of the Genesis of Metamorphic Zircons 131 0.31, which is quite consistent with the metamorphic zircons. This is confirmed by the
isotopic age of all the crystals, which is close and approximately 2.1 Ga (Pystin and
Pystina 2015b, Pystina and Pystin 2002). Our studies show that accessory zircons from rocks of different granitoid com-
plexes in the northern part of the Subpolar Urals, which have different geological
positions and isotopic age, differ in the set of morphotypes, their quantitative ratios and
geochemical features (Pystina et al. 2017). Recently we studied morphological features
of zircons from the granites of Nikolayshor (PR1), Kozhim (PF2–3), Badyayu (RF3–
V) and Yarota (RF3–V) Massifs. Accessory zircons are very diverse in form, character
of zonality, presence of inclusions, color, degree of metamictism and other features. All
the main morphological types of zircons according to I.V. Nosyrev (Nosyrev et al. 1989) were observed: zirconium, hyacinth, spear-shaped, torpedo-shaped and citrolite. All of the above morphological types can relate to the generation of zircons of either
synpetrogenic or superimposed genetic types. In the granites of the Nikolashor Massif hyacinth, spear-shaped, torpedo-shaped,
and detrital zircons were found. Granitoids of the Kozhimsky, Badyayusky and
Yarotsky Massifs are characterized by the presence of three morphotypes of zircons,
but if in the granitoids of the Badyayu and Yarota Massifs they are similar (zircon,
hyacinth and torpedo-shaped morphotypes), while in the rocks of the Kozhim Massif,
they are pattern, torpedo-shaped, torrent, and the beginning of torpedo-shaped mor-
photypes. Common to the granitoids of all the massifs is one morphotype – torpedo. 2
Results and Discussion The spear-shaped zircon is found only in the rocks of the Nikolashor Massif. There are
also detrital zircons, which are absent in the granitoids of other massifs. Granites of the
Kozhim Massif are distinguished from other granitoids by the presence of zircon of the
citrolite morphotype. The presence of this type of zircons is a sign of the metasomatic
(or metamorphic) processing of rocks. In contrast to the granites of the Badyayu and
Yarota Massifs, they lack hyacinth type zircons. The Th/U ratio in zircons from the granitoids of the northern part of the Subpolar
Urals – Nikolashor, Kozhim, Khatalambo-Lapchin and Lapchavozh Massifs, is on
average 0.73; 0.61; 0.51; 0.79, respectively. These values are maintained and observed
in all zircons of the studied granitoids. In some cases, in zircons from the granitoids of
the Khatalambo-Lapchin Massif, the Th/U values are knocked out of the overall pic-
ture, amounting to 0.22 and 0.15, which is not at all characteristic of magmatic zircons. If we take into account that the age of these zircon crystals obtained by the U-Pb
SHRIMP-II method is 703.9 ± 8 Ma and 795 ± 41 Ma, and the rest of the zircons is
550–580 Ma, we can assume that the formation of ancient zircons The early stages of
granite formation and the increased Th/U ratio are explained by the subsequent
metamorphism of early generation granites. References Hoskin PWO, Schaltegger U (2003) The composition of zircon and igneous and metamorphic
petrogenesis. Rev Miner Geochem 53:27–62 Krasnobayev A (1986) Zircon as an indicator of geological processes. Nauk, Moscow, 152 Krasnobayev A (1986) Zircon as an indicator of geological processes. Nauk, Moscow, 152 p
Nosyrev IV, Robul VM, Esipchuk KE, Orsa VI (1989) Generation analysis of accessory zircon. Nauka Press, Moscow 203 p Nosyrev IV, Robul VM, Esipchuk KE, Orsa VI (1989) Generation analysis of accessory zircon. Nauka Press, Moscow 203 p Pystin A, Pystina J (2015a) The early precambrian history of rock metamorphism in the Urals
segment of crust. Int Geol Rev 57(11–12):1650–1659 Pystin AM, Pystina YI (2008) Metamorphism and granite formation in the Proterozoic-
Earlypaleozoic history of the formation of the Polar Ural segment of the earth’s crust. Lithosphere (6):25–38 Pystin AM, Pystina YI (2015b) The Archean-Paleoproterozoic history of metamorphism of rocks
of the Ural segment of the earth’s crust. Works of the Karelian Research Center of the Russian
Academy of Sciences, no 7, Geology of Dokembriya, pp 3–18 Pystina YI, Denisova YV, Pystin AM (2017) Typomorphic signs of zircons as a criterion for the
dissection and correlation of granitoids (by the example of the northern part of the Subpolar
Urals). Bulletin of the Institute of Geology of Komi Scientific Center of the Ural Branch of
the Russian Academy of Sciences, no 12, pp 3–15 Pystina Y, Pystin A (2002) Zircon chronicle of the Ural Precambrian. UrD RAS Press,
Ekaterinburg, 167 p Rubatto D (2002) Zircon trace element geochemistry: partitioning with garnet and the link
between U-Pb ages and metamorphism. Chem Geol 184:123–138 Skublov SG, Lobach-Zhuchenko SB, Guseva NS, et al (2009) Distribution of rare-earth and rare
elements in zircons from miaskite lamproites of the Panozersky complex of central Karelia. Geochemistry (9):958–971 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. 3
Conclusions Thus, we have to state the validity of the fact that “the only obvious systematic
difference between the magmatic and metamorphic zircon is the Th/U ratio …”
[Hoskin and Schaltegger 2003, p. 48]. It allows not only to distinguish between Y. Pystina and A. Pystin 132 magmatic zircons from metamorphic, but taking into account the morphological fea-
tures of individual crystals and isotopic age dating, it is more reliable to restore the
history of the formation of specific metamorphic and igneous complexes. magmatic zircons from metamorphic, but taking into account the morphological fea-
tures of individual crystals and isotopic age dating, it is more reliable to restore the
history of the formation of specific metamorphic and igneous complexes. Acknowledgements. This work was supported by the Basic Research Program of the Russian
Academy of Sciences No. 18-5-5-19. Acknowledgements. This work was supported by the Basic Research Program of the Russian
Academy of Sciences No. 18-5-5-19. References If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction The problem of processing of aluminum ores is related to the fact that bauxite-forming
minerals are characterized by similar values of density, dispersion and fine mutual
intergrowths of individuals, non-recoverability of many valuable microelements. Therefore, the development and improvement of methods for the enrichment and
processing of bauxite remains highly relevant (Borra et al. 2015; Borra et al. 2016;
Davros et al. 2016). In terms of their iron index, the studied Timan bauxites are
subdivided into three mineral types: hematite-boehmite, hematite-berthierine-boehmite,
and berthierine-boehmite (Vakhrushev 2011, 2012; Vakhrushev et al. 2012). The
results of our studies showed that in these bauxites in the state of endocrypt scattering
there are a lot of valuable elements-impurities, which extraction by modern tech-
nologies is either time consuming or not yet possible. Our experiments on heating of
bauxites in combination with their irradiation with high-energy electrons lead to a
change in the phase composition of bauxites and, as a result, to the improvement of
their technological properties (Rostovtsev 2010; Kotova et al. 2016). © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 133–135, 2019.
https://doi.org/10.1007/978-3-030-22974-0_31 Phenomenon of Microphase Heterogenization
by Means of Endocrypt-Scattered Impurity
of Rare and Noble Metals as a Result
of Radiation by Accelerated Electrons
of Bauxites I. Razmyslov1(&), O. Kotova1, V. Silaev1, and L. A. Gomze2 1 Institute of Geology Komi SC UB RAS, Syktyvkar, Russia
razmyslov-i@mail.ru
2 University of Miskolc, Miskolc, Hungary Abstract. During radiation-thermal transformation of Timan ferrous bauxites
we discovered a previously unknown phenomenon of microphaseheteroge-
nization, which can contribute to the extraction of many valuable impurities
during processing of relatively poor quality bauxite raw. Keywords: Bauxite Radiation-thermal transformation
Microphase heterogenization Profitability of bauxite raw processing Keywords: Bauxite Radiation-thermal transformation
Microphase heterogenization Profitability of bauxite raw processing 2
Methods and Approaches We carried out experiments with thermal (heating to 500 and 600 °C with quadruple
exposure by 60 min) and radiation-thermal (heating to 500–600 °C with double
exposure by 20 min with irradiation by an electron beam with energy of 2.4 MeV 134 I. Razmyslov et al. using ILU-6 industrial unit at the Institute of Nuclear Physics of the Siberian Branch of
the Russian Academy of Sciences) modification of Timan iron bauxites. References Borra CR, Mermans J, Blanpain B, Pountikes YB, Gerven T (2016) Selective recovery of rare
earths from bauxite residue by cjmbination of sulfation, roasting and leaching. Min Eng
92:151–159 Borra CR, Pontikes Y, Binnemans K, Gerven T (2015) Leaching of rare earts from bauxite
residue (red mud). Min Eng 76:20–27 Davros P, Balomenos E, Panias D, Paspaliaris I (2016) Selective leaching 0f rare earth elements
from bauxite residue (red mud). Hydrometallurgy 164:125–135 Davros P, Balomenos E, Panias D, Paspaliaris I (2016) Selective leaching 0f rare earth elements
from bauxite residue (red mud). Hydrometallurgy 164:125–135 Kotova OB, Razmyslov IN, Rostovtsev VI, Silaev VI (2016) Radiation-thermal modification of
ferruginous bauxites during processing. Enrich Process 4:16–22 (in Russian) Kotova OB, Razmyslov IN, Rostovtsev VI, Silaev VI (2016) Radiation-thermal modification of
ferruginous bauxites during processing. Enrich Process 4:16–22 (in Russian) Rostovtsev VI (2010) Theoretical foundations and practice of using electrochemical and radiation
(accelerated electrons) effects in the processes of ore preparation and enrichment of mineral
raw materials. Vestnik of the Chita State University, vol 8, pp 91–99. (in Russian) Rostovtsev VI (2010) Theoretical foundations and practice of using electrochemical and radiation
(accelerated electrons) effects in the processes of ore preparation and enrichment of mineral
raw materials. Vestnik of the Chita State University, vol 8, pp 91–99. (in Russian) Vakhrushev AV (2012) Crystal chemistry of bauxite minerals from the Vezhayu-Vorykvinskoe
deposit. Structure, substance, history of the lithosphere of the Timan-Northern Ural segment. Geoprint, Syktyvkar, pp 32–34. (in Russian) Vakhrushev AV, Lyutoev VP, Silaev VI (2012) Crystal-chemical features of ferrous minerals in
bauxite at the Vezhayu-Vorykvinskoe deposit (Middle Timan). IG Komi Science Center UB
RAS, pp 14–18. (in Russian) Vakhrushev AV, Lyutoev VP, Silaev VI (2012) Crystal-chemical features of ferrous minerals in
bauxite at the Vezhayu-Vorykvinskoe deposit (Middle Timan). IG Komi Science Center UB
RAS, pp 14–18. (in Russian) Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. Phenomenon of Microphase Heterogenization Acknowledgements. This research was supported by UB RAS project 15-18-5-44 and project
AAAA-A19-119031390057-5 “The main directions of integrated assessment and effective use of
geo-resources in the Timan-North Ural-Barents Sea region”. 3
Results and Discussion Heating up to 500–600 °C with exposure to 60 min without irradiation led to almost
complete dehydration of Al and Fe oxyhydroxides with the formation of cAl2O3 (spi-
nelide with a defective structure) and hematite with relatively small alteration of
structure and magnetic properties of the rocks. The gross chemical composition of the
annealed samples remained almost unaltered, and the content of trace elements
increased markedly in% to the original: Cu by 70–250; Zn by 20–25; Cd by 20–380; Zr
by 2–20; Nb by 15–20; Sc by 25–40; Y by 35–70; Ce by 25–60; Nd by 1–10. Obviously,
the latter is conditioned bya significant thermal dehydration of the studied bauxites. The radiation-thermal treatment of ferrous bauxites led not only to dehydration of
the original minerals, but also to chemical disproportionation of the original substance,
its radical magnetic restructuring resulted from the presence of strong magnetic phases
(maghemite, magnetite) and the formation of valuable trace elements of many new
minerals with individuals varying in shape from isometric to needle-like and in size
from submicronic to 0.5 mm due to endocrypt scattered impurity. The newly formed
phases include native metals (Au, Pb, Al, Cu, Zn); sulfides (pyrite, galena); oxides of
Sn, Ta, Nb, Zr, lanthanides; silicates (zircon, kaolin); rare sulphates, etc. Thus, the
heating, combined with irradiation with high-energy electrons, resulted not only in
transformation of primary minerals in the ferrous bauxites, but also in microphase
heterogenization with the formation of new minerals (Fig. 1). It can be assumed that
this kind of transformation can contribute to extraction of many valuable impurities and
increase the profitability of processing of relatively low-quality bauxite raw. Fig. 1. Micro pocket segregations of Ce-Gd oxides in radiation-thermally modified Timan
bauxites Fig. 1. Micro pocket segregations of Ce-Gd oxides in radiation-thermally modified Timan
bauxites Phenomenon of Microphase Heterogenization 135 References If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction The development of the methods of flotation, gravity and metallurgic processing of
gold bearing ores and man-made products today is aimed at increase of the recovery of
the valuable product (Pavlov 2005; Pavlov et al. 2015; Meimanova and Nogayeva
2014; Bogdanovich et al. 2013; Tselyuk and Tselyuk 2013; Algebraistova et al. 2017;
Amdur et al. 2015). There are almost no technologies for comprehensive use of pro-
cessing tails to produce an additional marketable product both for task of compre-
hensive use of processing tails for production of foamed resources for ceramic items
production with associated gold recovery to the ferrosilicium matrix is vital. Gold Extraction to Ferrosilicium, Production
of Foam Silicate from Processing Tails
of the Olimpiada Mining and Processing
Complex Gold Processing Plant
(Russia, Krasnoyarsk Territory) A. Sazonov1(&), V. Pavlov2, S. Silyanov1, and E. Zvyagina1 1 Siberian Federal University, Krasnoyarsk, Russia
Sazonov_am@mail.ru
2 Special Design and Technology Bureau “Science”, Krasnoyarsk, Russia 2 Special Design and Technology Bureau “Science”, Krasnoyarsk, Russia Abstract. The paper describes the studies of processing tails of the Olimpiada
Mining and Processing Complex with the methods of chemical, mineralogical,
electronic microscope, deep reductive melting with division of the melt into a
silicate and metal parts. It is demonstrated that 85% of the processing tails
consist of the oxides: SiO2, CaO, Al2O3, Fe2O3, H2O and CO2. The distribution
of gold and silver is provided by the size classes of the initial blend, after
melting of which in reduction conditions re-distribution of gold to the metal
phase of the melt occurs. The silicate part of the melt when released into water in
the “thermal shock” mode forms a light porous X-ray amorphous material “foam
silicate”, which also serves as a resource for stable chemical composition for
production of a wide range of import substituting ceramic materials. Keywords: Pyrometallurgy Processing tails Ferrosilicium Gold extraction © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 136–139, 2019.
https://doi.org/10.1007/978-3-030-22974-0_32 3
Samples The input material for pyrometallurgic extraction of gold is represented by the tails of
the gold processing plant of the Olimpiada Mining and Processing Complex, which is
the leader in Russia in terms of the ore processing volumes and marketable gold
production. The gold-antimony and gold-arsenic ores of the Olimpiada deposit have a
complex mineral composition and are extremely refractory for gold recovery from
them. Most of gold is in the form of thin dissemination in sulfides (Kirik et al. 2017;
Novozhilov et al. 2014). No more than 50% of gold is extracted by direct cyanation. At
the gold processing plant the flotation concentrates of the ores are exposed to bioox-
idation using the BIONORD® technology and subsequent leaching to release gold
capsuled in sulfides. The processing tails have loose consistence, mainly sand-aleuritic-clayey size, with
the content of (−0.071) mm fraction of about 25%. The chemical composition is
dominated by SiO2, CaO, Al2O3, H2O and CO2, the share of which is about 90 wt.%. The concentrations of S are 0.69%, C—2.35%, Sb—0.11%, As—0.19%, and Ag
— < .1%. The gold grade in individual samples as per the fire assay data varies from 0.2
to 0.97 g/t, with the average metal grade in the bulk sample received for study of 0.6 g/t. The main minerals of tails are quartz, calcite and stratified silicates making 98–99%. Sulfides, oxides, hydroxides and sulfates of iron, arsenic, antimony and tungsten are the
impurities. In addition to the native highcarat gold, aurostibite (AuSb2) and jonassonite
(AuBi5S4) are present in single grains. Copper, antimony and mercury impurities are
noted in some gold particles. Native gold particles are less than 0.045 mm in size (90%). Most of gold is noted in the form of micron inclusions in sulfides, quartz, carbonate and
micas. Successive chemical leaching in the tail material using the method (Antropova
et al. 1980) identified about 20% of mobile gold forms (water-soluble, sorbed, ferri-
forms) and sulfide and telluride forms * 15%. 2
Methods The paper proposes the pyrometallurgic approach to the solution of the problem of
comprehensive wasteless processing of man-made gold-bearing resources with the
method of deep reductive melting with division of the melt into the deferrized silicate Gold Extraction to Ferrosilicium, Production of Foam Silicate 137 part with its further chilling in the thermal shock mode (Pavlov et al. 2015) and the
metal part with associated re-distribution of gold in the ferrosilicium matrix. The
specimens of the silicate part of the melt were prepared for study using the powder
technology; the metal specimens were covered with epoxy resin with subsequent
polishing. The chemical phase composition was studied with the use of an X-ray
fluorescent
S2
RANGER
analyzer
and
the
scanning
electronic
microscope
(SEM) Hitachi S-3400N with EMF Bruker. 4
Results In the process of the experiment the flotation tails sample with the weight of 400 g was
mixed with lime (Ca(OH)2) and brown coal. The blend was exposed to reductive
melting at the temperature of 1500–1550 °C, in the process of which melt division into
a metal and silicate parts occurred. The silicate part of the melt was poured into water A. Sazonov et al. 138 with production of foamed amorphous material (foam silicate). The recovery of foam
silicate was 150 g. The metal phase recovery was 26 g (4.4%). Phase composition of
the metal aggregate (wt.%): ferrosilicum (FeSi) 82.8; xifengite (Fe5S3) 8.97; wustite
(Fe0.974O) 0.79; wollastonite (CaSiO3) 4.24; calcic clinoferrosilite (Fe1.5Ca0.5(SiO3)2)
and elemental iron (Fe) 0.49. Chemical composition of the produced foam silicate (wt. %) – SiO2—43.7; TiO2—0.7; Al2O3—7.79; Fe2O3—0.19; MgO—2.22; CaO—42.5;
K2O—1.5; SO3—0.77; Cl—0.27, and metal phases: Si—23.5; Al—1.6; Fe—66.2; Mn
—2.22; Mg 0.47; Ca—1.97; S—0.42; As—0.68; Cl—0.14; P—1.0; Co—0.35; V—
0.28; Cu—0.21; Au—0.2; and Cr—0.18. The optical research of the metal alloy
showed non-uniform aggregate composition. Six individualized metal phases of more
complex composition have been identified in the iron and silicon alloy matrix. Gold is a
part of the alloy consisting of (wt.%): Au—0.25–5.11; Sb—0.4–0.7; Sn—0.57–3.30;
As—up to 9.47; Cu—4.51–32.07; Fe—59.8–33.8; Mn—7.2–1.33; Ga—0.24–9.38;
and Si—14.53–12.96. 5
Conclusions Therefore, silicate and metallic semi-finished products have been produced as the result
of deep reductive melting of processing tails: (1) ferrosilicium, which is a gold col-
lector; (2) foam silicate material as an additional product of the main production, can be
used for production of ceramic materials for different purposes. The use of the method
of pyrometallurgic processing of processing tails allows mitigating their adverse effect
on the environment. References Algebraistova NK, Makshanin AV, Burdakova EA, Markova AS (2017) Processing of precious -
metal raw materials in centrifugal devices. Non-ferrous Met 1:18–22 Amdur AM, Vatolin NA, Fyodorov SA, Matushkina AM (2015) Movement of disperse drops of
gold in porous bodies and oxide melts during heating. Rep Acad Sci 465(3):307–309 ntropova LV, Shuraleva AZ, Farfel LF, Aizenberg FM, Priyemov GA (1980) Forms of gold
occurrence in the rock. Explor Met Eng 136:5–21 Antropova LV, Shuraleva AZ, Farfel LF, Aizenberg FM, Priyemov GA (1980) Forms of gold
occurrence in the rock. Explor Met Eng 136:5–21
Bogdanovich AV, Vasilyev AM, Shneyerson YaM, Pleshkov MA (2013) Gold extraction from
stale processing tails of pyritic copper zinc ores Ore Process 5:38 44 occurrence in the rock. Explor Met Eng 136:5–21
Bogdanovich AV, Vasilyev AM, Shneyerson YaM, Pleshkov MA (2013) Gold extraction from
stale processing tails of pyritic copper-zinc ores. Ore Process 5:38–44 ogdanovich AV, Vasilyev AM, Shneyerson YaM, Pleshkov MA (2013) Gold extraction from
stale processing tails of pyritic copper-zinc ores. Ore Process 5:38–44 Kirik SD, Sazonov AM, Silyanov SA, Bayukov OA (2017) Study of disordering in the structure
of natural arsenopyrite with the X-ray diffraction analysis of polycrystals and nuclear gamma
resonance. J Siberian Federal Univ Eng Technol 10(5):578–592 Meimanova ZhS, Nogayeva KA (2014) Study of flotation processability of stale tails from the
Solton-Sary processing plant. Nauka i novye Tekhnologii 2:15–16 Novozhilov YI, Gavrilov AM, Yablokova SV, Arefyeva VI (2014) Unique commercial gold
antimony deposit Olimpiada in upper proterozoic terrigenous deposits. Ores Met 3:51–64 Pavlov VF (2005) Physical bases for the technology of production of new materials with set
properties on the basis of creation of the system for comprehensive use of man-made and
barren resources. Siberian Branch of the Russian Academy of Sciences, Novosibirsk Gold Extraction to Ferrosilicium, Production of Foam Silicate 139 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. Gold Extraction to Ferrosilicium, Production of Foam Silicate Pavlov MV, Pavlov IV, Pavlov VF, Shabanova OV, Shabanov AV (2015) Features of processes
of pyrometallurgic processing of polymetallic ores. Chem Benefit Sustain Dev 3:263–266 Pavlov MV, Pavlov IV, Pavlov VF, Shabanova OV, Shabanov AV (2015) Features of processes
of pyrometallurgic processing of polymetallic ores. Chem Benefit Sustain Dev 3:263–266
Tselyuk OI, Tselyuk DI (2013) Prospects of use of gold heap leaching for involvement into
commercial development of stale tails of the Eastern Siberia gold processing plants. In:
Proceedings of the Siberian Department of the Section of Earth Sciences of the Russian
Academy of Natural Sciences, vol 1, no. 42 pp 103–110 Tselyuk OI, Tselyuk DI (2013) Prospects of use of gold heap leaching for involvement into
commercial development of stale tails of the Eastern Siberia gold processing plants. In:
Proceedings of the Siberian Department of the Section of Earth Sciences of the Russian
Academy of Natural Sciences, vol 1, no. 42 pp 103–110 Karpinsky Russian Geological Research Institute, St. Petersburg, Russia
Rimma_Brodskaya@vsegei.ru Abstract. The paper is supposed to discuss one of the most important techno-
logical properties of mineral aggregates: their ability to disintegrate, to be
destroyed for the subsequent redistribution and extraction of a useful component. The strength of a mineral aggregate is determined by the strength of accretion of
composing individuals and depends on many factors, including the energy
(kinetics) of formation conditions and the subsequent transformation conditions. The energy of the boundary of accretion of mineral grains in aggregate cannot be
directly measured since currently, there are no reliable and proved methods for
measuring the surface energy of solids. It is only possible to calculate quantity,
proportional to the surface energy of the fusion of mineral grains – this is the
calculation of the atomic density of the surface, parallel to the boundary of
accretion of a pair of mineral grains. The methodology and calculation of this
characteristic have been developed and published. The use and obtaining of
results are possible at the stage of preliminary mineralogical study by methods of
geometrical analysis of the structure of the mineral aggregate. The effectiveness of
the proposed method is determined by the knowledge of the quality and features
of destruction of the aggregates using different physical methods of disintegra-
tion: traditional mechanical, ultrasonic, electro-pulse, electro-hydraulic. Keywords: Technological properties Mineral aggregate
Aggregate disintegration Repartition Strength Accretion boundaries
Energy of boundaries Keywords: Technological properties Mineral aggregate
Aggregate disintegration Repartition Strength Accretion boundaries
Energy of boundaries References If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Predictive Assessment of Quality of Mineral
Aggregates Disintegration
S. Shevchenko, R. Brodskaya(&), I. Bilskaya, Yu. Kobzeva,
and V. Lyahnitskaya
Karpinsky Russian Geological Research Institute, St. Petersburg, Russia
Rimma_Brodskaya@vsegei.ru 1
Introduction The mineral aggregate is considered as a structured system consisting of mineral
individuals and the accretion boundaries between them. The formation of the aggregate
results from cooperative thermodynamic processes of synthesis of substances, corre-
sponding to the composition of minerals, as well as the crystallization of mineral
individuals and the formation of crystal-grain boundaries, their aggregation. The
aggregation or accretion of mineral individuals with each other is carried out along the
forming boundaries of accretion. The process of aggregation of grains in the mineral
aggregate may somewhat lag behind the synthesis and crystallization of individuals,
but their parallel development is also possible. The accretion boundaries do not always © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 140–142, 2019. https://doi.org/10.1007/978-3-030-22974-0_33 Predictive Assessment of Quality of Mineral Aggregates 141 correspond to the boundaries of the crystallization of individuals, since they are formed
in a different energy situation. correspond to the boundaries of the crystallization of individuals, since they are formed
in a different energy situation. 2
Methods and Approaches The accretion boundaries of mineral individuals represent a certain transition zone
between crystal lattices of accreted grains. The orientation of the accretion boundaries
with respect to the symmetry of crystalline lattices of mineral grains depends on the
local energy potentials during the formation and transformation of the mineral indi-
viduals and the aggregate. Local thermodynamic potentials of the forming mineral
system directly affect the safety of an individual through the energy of its boundary, the
choice of the orientation of the boundary relative to the symmetry of its crystal lattice
and the orientation of the accretion boundary with the nearest stable grain. Thus, a
compromise boundary is established for the accretion of each pair of mineral indi-
viduals, which is stable in a specific range of thermodynamic forces and flows. During evaluation of sequence and quality of opening of the accretions, it is nec-
essary to take into account the distribution of mineral individuals in the aggregate
which possess a very perfect cleavage. It is possible to assess the degree and quality of opening of the accretions at the
stage of preliminary mineralogical analysis when studying the structure of the mineral
aggregate in thin sections using geometric analysis methods. 3
Results and Discussion The boundaries of mineral individuals, as well as the accretion boundaries, possess a
certain stability reserve, ensuring their functioning in a certain range of thermodynamic
and mechanical parameters. The stability margin of the boundaries is the wider, the
higher the kinetics of the cooperative processes that form them, their genesis. The
accretion boundaries of mineral grains are the more stable in the field of destructive
forces, the more surface energy they accumulate. The stability of the accretion boundary of mineral grains depends, in addition to the
above said, on the degree of its balance by the value of energy saturation of each of the
mineral grains in the accretion region. The open boundaries of the mineral grains of accretions after relaxation of the
crystal lattice, as well as remaining unopened accretions, have different elemental
composition, amount of energy, and different floatability. S. Shevchenko et al. S. Shevchenko et al. The strength of mineral aggregates is understood as resistance to destructive forces
on the system of mineral grains forming the aggregate due to the presence of their
“accretion” surfaces. The accretion boundaries of the mineral grains can be balanced or unbalanced by
value of energy of accretion surfaces of individuals. The accretion of grains on surfaces, which atomic density or crystallographic
indices correspond to the edges of habit forms, will collapse faster and earlier by the
same destructive force, than the accretion of energy-intensive boundaries with a low
atomic density and high crystallographic indices. Brodskaya RL, Makagonov EP (1990) Determination of the spatial orientation of crystalline
individuals in microstructural analysis and prospects for its use in stereometric petrography.
Notes All-Union Mineral Soc 119(4):84–93 Brodskaya RL, Bil’skaya IV, Lyakhnitskaya VD, Markovsky BA, Sidorov EG (2007)
Boundaries of accretions between mineral individuals in an aggregate. Geol Ore Deposits
49(8):669–680 4
Conclusions The accretion boundaries determine the grain system as a mineral aggregate and
determine its stability, as both the strength of the accretions of mineral grains, and the
strength of the mineral aggregate as a whole. 142 2
Methods and Approaches A wide range of methods were used for the current research: the analysis of the earlier
conducted researches, the mathematical and physical simulation, determination of
dependences, the calculations and the control, the experimental studies and measure-
ments in accordance with the conventional standard. For the experience, we used a section of a rectangular shaped heap leach pile
5 5 m size. We have installed an air injection pipe near a homogeneous low-
permeable layer. Assuming that the bottom and sides are sealed to air leakage, we used
the SVAirFlow software to simulate the air/oxygen flow through a uniform anti-
filtration layer. In addition, the temperature inside the mass is constant and equal to
20 °C. The air supply pressure through the pipeline is 121 kPa, and the upper part has
an atmospheric pressure of 101 kPa. References Brodskaya RL, Makagonov EP (1990) Determination of the spatial orientation of crystalline
individuals in microstructural analysis and prospects for its use in stereometric petrography. Notes All-Union Mineral Soc 119(4):84–93 Brodskaya RL, Bil’skaya IV, Lyakhnitskaya VD, Markovsky BA, Sidorov EG (2007)
Boundaries of accretions between mineral individuals in an aggregate. Geol Ore Deposits
49(8):669–680 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Development of Methods for Anti-filtration
Formations Destruction Inside a Heap
Leach Pile
H. Tcharo(&), M. Koulibaly, and F. K. N. Tchibozo
Department of Mineral Developing and Oil&Gas Engineering,
Engineering Academy, RUDN University, Moscow, Russia
honoretcharo@yahoo.com 1
Introduction The presence of low permeable soils is one of the most difficult problems of metal
extraction during heap leaching (Ozhogina et al. 2017; Vorobyov et al. 2017). The development of new models will reduce the degree of their negative impact on
the intensity of heap leach process. Development of Methods for Anti-filtration
Formations Destruction Inside a Heap
Leach Pile H. Tcharo(&), M. Koulibaly, and F. K. N. Tchibozo Department of Mineral Developing and Oil&Gas Engineering,
Engineering Academy, RUDN University, Moscow, Russia
honoretcharo@yahoo.com Abstract. This article discusses the new technical solutions that increase
the restoration of the quality of pregnant solutions flowing out from the heap
leach pile. Keywords: Anti-filtration formations Heap pile Pipe Sprinklers © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 143–145, 2019.
https://doi.org/10.1007/978-3-030-22974-0_34 H. Tcharo et al. The second technical solution, which also allows to increase the intensification of
heap leach process is the displacement of spraying devices in the direction of weakly
affected by technological solutions places. The second technical solution, which also allows to increase the intensification of
heap leach process is the displacement of spraying devices in the direction of weakly
affected by technological solutions places. Fig. 1. Model showing the impact of injecting air/oxygen Fig. 1. Model showing the impact of injecting air/oxygen 4
Conclusions According to our study results, we can conclude that supplying the air under much
higher pressure than the atmospheric one in the places where anti-filtration layers are
formed, along with the displacement of the sprinklers, will enhance more seepage of
pregnant solutions through a heap leach pile. 3
Results and Discussion The results of the model are shown in Fig. 1. We’ve noticed that the air pressure in the
whole low-permeable layer varied between 109 and 120 kPa. This means that the pores
are slightly wider open. 144 H. Tcharo et al. Ozhogina EG, Shadrunova IV, Chekushina TV (2017) Mineralogical rationale for solving
environmental problems of mining regions. Gornyi zhurnal 11:105–110
Vorobyov AE, Lyashenko VI, Tcharo H, Vorobyev KA (2017) Efficiency increase for gold-
bearing ore deposits with respect to the influence of sulfide surface nanobarrier on metal
adsorption. Sci Tech J “Metall Min Ind” 7:28–36 Microtomographic Study of Gabbro-Diabase
Structural Transformations Under
Compressive Loads
L. Vaisberg1(&) and E. Kameneva2
1 «Mekhanobr-Tekhnika» REC, St. Petersburg, Russia
gornyi@mtspb.com
2 Petrozavodsk State University, Petrozavodsk, Russia Microtomographic Study of Gabbro-Diabase
Structural Transformations Under
Compressive Loads
L. Vaisberg1(&) and E. Kameneva2
1 «Mekhanobr-Tekhnika» REC, St. Petersburg, Russia
gornyi@mtspb.com
2 Petrozavodsk State University, Petrozavodsk, Russia Microtomographic Study of Gabbro-Diabase
Structural Transformations Under
Compressive Loads L. Vaisberg1(&) and E. Kameneva2 1 «Mekhanobr-Tekhnika» REC, St. Petersburg, Russia
gornyi@mtspb.com
2 Petrozavodsk State University, Petrozavodsk, Russia Abstract. Transformations of the porous space structure for gabbro-diabase
under compressive loads is studied. Quantitative assessment of respective
structural parameters for the pore space is ensured through the application of
X-ray computer microtomography, enabling visualization of the internal three-
dimensional structure of each sample and a detailed quantitative analysis of the
pore space structure for both separate sections and the entire sample volume. Differences in the number, sizes, shapes, connectivity and spatial distribution of
pores are established. It is shown that, when the sample is destroyed, the structure
of the pore space in its fragments is transformed as follows: intracrystalline pores
are partially closing, with the simultaneous emergence of new pores of large
capillary sizes, concentrated in the cracks. In terms of their structure, these cracks
represent a system of interconnected pores containing micron-size mineral
particles. Keywords: Computer microtomography Gabbro-diabase Pore space Keywords: Computer microtomography Gabbro-diabase Pore space © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 146–151, 2019.
https://doi.org/10.1007/978-3-030-22974-0_35 References Ozhogina EG, Shadrunova IV, Chekushina TV (2017) Mineralogical rationale for solving
environmental problems of mining regions. Gornyi zhurnal 11:105–110 Ozhogina EG, Shadrunova IV, Chekushina TV (2017) Mineralogical rationale for solving
environmental problems of mining regions. Gornyi zhurnal 11:105–110 Vorobyov AE, Lyashenko VI, Tcharo H, Vorobyev KA (2017) Efficiency increase for gold-
bearing ore deposits with respect to the influence of sulfide surface nanobarrier on metal
adsorption. Sci Tech J “Metall Min Ind” 7:28–36 145 Development of Methods for Anti-Filtration Formations Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction In the current theory for the mechanism of disintegration of rocks, destruction is a
process that develops in time. The formation and development of microscopic failures
begin upon application of a loading force, either dynamic or static, and resume for the
entire period the rock remains under load until fracture (Zhurkov 1980; Krivtsov 2007). The existing works on rock destruction generally identify the most probable
structural elements, along which the destruction processes tend to develop. These are
pores, tiny fractures, intergrowth boundaries of mineral phases and intergrain bound-
aries. In this regard, porosity, understood as the sum of all cavities enclosed in the rock,
including pores, pore channels, and tiny fractures, becomes a useful feature, linking the
strength of a rock with the defects in its structure. The physical or total porosity of a
rock, determined by calculation using known values of mineral and bulk densities,
enables only indirectly assessing the transformations occurring in the rock microstruc-
ture during destruction. Unbiased data may only be obtained if the structure of the pore © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 146–151, 2019. https://doi.org/10.1007/978-3-030-22974-0_35 Microtomographic Study of Gabbro-Diabase Structural Transformations 147 space is taken into account, including its dimensions, pore shapes, connectivity and
spatial orientation (Romm 1985). The purpose of this work is to study the transformations occurring in the
microstructure of a rock during destruction. X-ray computer microtomography was
selected as the method for the quantitative assessment of respective pore space struc-
tural parameters of rocks at the microscopic level. 2
Methods and Approaches X-ray tomography (X-ray micro-CT) is a non-destructive method for studying the
internal structure of solid materials, based on the dependence of the linear coefficient of
X-ray radiation attenuation on the chemical composition and density of the substance
analyzed. Computer processing of shadow projections obtained by x-ray scanning of
samples allows visualizing the internal three-dimensional structure of each sample and
performing a detailed analysis of its morphometric and density characteristics both at
separate sections and throughout the entire sample volume, obtaining quantitative
values of respective parameters. The non-destructive nature of the method is an
important advantage of X-ray microtomography, as it enables subsequent application of
the same samples for other types of analysis, in particular, for establishing their strength
characteristics. A gabbro-diabase sample of cylindrical shape (L = d) without visible defects was
prepared for the studies. The gabbro-diabase sample was characterized by a massive
texture and a uniform medium-grained structure. The size of mineral grains was up to
2–3 mm. The experiment included X-ray tomography of the gabbro-diabase sample with
identification of the following parameters of the pore space: dimensions, shape, volume,
specific surface, pore connectivity, spatial orientation, distribution, and pore density,
both in separate tomographic Sections (2D system) and in the entire volume of the
sample (3D system). The sample was then subjected to axial loading using a manual
hydraulic press until fracture. The loading force was 192 MPa, the lateral pressure was
1 atm, at t = 20 °C, the loading time was 121 s. The resulting fragment was then
subjected to x-ray tomography. The tomographic studies were carried out using SkyScan-1172 (Belgium) with
resolutions of 0.5 to 27 lm. In the experiments, the samples were carefully oriented on
the table with respect to the optical axis of the instrument; the tube was supplied with
100 mA and 100 kV; the X-ray power was 90 W. The pixel size at the maximum
magnification (nominal resolution) was 3.9 lm, which generally allows identifying
pores of 4 lm or larger. The table with the sample was rotated by 360° in 0.25
increments. The subsequent reconstruction work was performed using Nrecon, CTan,
and CTvol SkyScan software. 148 L. Vaisberg and E. Kameneva 3
Results and Discussion The original sample contains crystals of 0.2–2.0 mm, represented by plagioclase
(41.5%), actinolite (47.0%), quartz (3.6%), sphene (5.7%) and biotite (2.2%). The porosity of the initial sample is 0.7%. The pores in the sample volume are
unevenly distributed. Plagioclase crystals have the highest porosity that is 4.1 to 4.9
times higher than the value for the entire sample (2.9–3.4% vs 0.7%). In quartz,
actinolite and biotite crystals, only individual pores are observed. The concentration of pores in the original sample is 84.55 mm−3. The largest pore size is 32–34 lm. In quantitative terms, pores of up to 10 lm
prevail. The largest pore size is 32–34 lm with their median value of 5.6 lm. In
quantitative terms, pores of up to 4–5 lm (80.7%) prevail in this group. 3 In the fragment after fracture, the total pore concentration increases to 163.9 mm−3,
super-capillary pores of 179 to 180 lm are observed, and the porosity increases to
1.8% (Table 1). Table 1. Results of microtomography (3D-system)
Parameters
Values
Original sample Fragment after sample fracture
Pore fraction in sample volume, % 0.7
1.8
Concentration of pores, mm−3
84.55
163.9
Largest pore size, lm
32–34
178–180
Pore connectivity, %
2.81
28.88 Table 1. Results of microtomography (3D-system) Pore sphericity decreases from small pores to large pores. Pores of 4–5 lm are
close to spherical in shape. Pores of 110–180 lm have elongated shapes, with their
sphericity not exceeding 0.15 to 0.3. It can be seen on the tomographic sections (Fig. 1) that the fragments formed upon
sample fracture contain incomplete cracks (not leading to the formation of fracture
surfaces) with the length of L = 7–8 mm and width of h = 40–150 lm (for main
cracks) and of h = 1–10 lm (for feather cracks). The study of the structure of the cracks shows that these are linear-plane sections
consisting of interconnected cavities containing micron-size mineral particles (Fig. 2). The availability of these cracks satisfactorily explains the increase in pore con-
nectivity. In the original sample, the pore connectivity is 2.81%; in the fractured
fragment, it is an order of magnitude higher (27.88%). The increase in connectivity
indicates the association of small pores into larger pores. Microtomographic Study of Gabbro-Diabase Structural Transformations 149 Fig. 1. Cracks in Gabbro-Diabase sample fragments (2D System) 1 m—Plagioclase; 2 m—
Actinolite, 3—Sphene Fig. 1. L. Vaisberg and E. Kameneva L. Vaisberg and E. Kameneva L. Vaisberg and E. Kameneva At the same time, a comparative tomographic study of the most porous mineral
component, the plagioclase crystal, shows a decrease in pore concentration under
compressive loads (Fig. 3). The plagioclase crystal porosity values are 3.23% for the
original sample and 1.11% for the fractured fragment, which indicates that the
intracrystalline pores are closing. Not only the finest pores, but also the larger ones are
joined: the maximum pore size in the plagioclase crystal is 32 lm in the original
sample and 16 lm in the fragment. Porosit
Dmax –
ty – 1,11%
16 um
Porosity – 3,23
Dmax – 32 um
%
Fig. 3. Pores (white) in Plagioclase crystal in original sample (a) and in its fractured fragment
(b) in 2D system Fig. 3. Pores (white) in Plagioclase crystal in original sample (a) and in its fractured fragment
(b) in 2D system The increase in the number of pores in the volume of the fragment formed after
fracture, with the closing of intracrystalline pores, suggests that the newly formed pores
are mainly concentrated in the cracks. In terms of their structures, these cracks rep-
resent a system of interconnected pores containing micron-size mineral particles. Acknowledgements. The study was supported by the grant of the Russian Science Foundation
(project No. 17-79-30056). 3
Results and Discussion Cracks in Gabbro-Diabase sample fragments (2D System) 1 m—Plagioclase; 2 m—
Actinolite, 3—Sphene Fig. 2. Structure of cracks Fig. 2. Structure of cracks 150 4
Conclusions When a gabbro-diabase sample is fractured under the action of a compressive load, the
structure of the pore space is transformed in the fragments formed as follows:
intracrystalline pores are partially closing, with the simultaneous emergence of new
pores of large capillary sizes, concentrated in the cracks. Acknowledgements. The study was supported by the grant of the Russian Science Foundation
(project No. 17-79-30056). Microtomographic Study of Gabbro-Diabase Structural Transformations 151 Krivtsov AM (2007) Deformation and fracture of solids with microstructure, Fizmatlit, Moscow
Romm YeS (1985) Structural models of pore space of rocks, Nedra, Leningrad
Zhurkov SN (1980) On the question of the physical nature of the strength. J Solid State Phys 22
(11):13–15 Microtomographic Study of Gabbro-Diabase Structural Transformations Process Mineralogy as a Basis
of Molybdoscheelite Ore Preparation 1 REC Mekhanobr-Tekhnica, St. Petersburg, Russia
gornyi@mtspb.com
2 Moscow State University, Moscow, Russia Abstract. The Tyrnyauz ore field (Big Tyrnyauz) is one of the largest and most
geologically complex deposits of tungsten and molybdenum. The main valuable
mineral of the deposit is molybdoscheelite, a representative of the sheelite—
powellite isomorphic series, as well as molybdenite and respective accessory
minerals. The main problem in the processing of ores of the Tyrnyauz ore field
consists in the high variability of mineral associations of its rocks, including the
availability of ores and nonmetallic minerals with similar physical and chemical
properties, as well as in the availability of ores of various natural types. Selective
mining and processing of various geological and industrial types of ores
with a wide use of vibrational technologies for the selective disintegration of
raw materials is a promising approach to the development of the Tyrnyauz
concentrator. Keywords: Molybdoscheelite Tungsten Molybdenum Disintegration
Separation Flotation Keywords: Molybdoscheelite Tungsten Molybdenum Disintegration
Separation Flotation References Krivtsov AM (2007) Deformation and fracture of solids with microstructure, Fizmatlit, Moscow
Romm YeS (1985) Structural models of pore space of rocks, Nedra, Leningrad
Zhurkov SN (1980) On the question of the physical nature of the strength. J Solid State Phys 22
(11):13–15 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 152–156, 2019.
https://doi.org/10.1007/978-3-030-22974-0_36 1
Introduction The Tyrnyauz ore field is located in the North Caucasus and is one of the largest and
most geologically complex deposits of tungsten, molybdenum and associated metals. The ore bodies of the Tyrnyauz deposit occur on the platform of Eljurta granites and
diabases. In the central part, these have undergone intensive metamorphism and con-
verted into amphibole-biotite hornfelses, overlain by numerous contact-metasomatic
and hydrothermal formations. The main valuable minerals of the deposit are molyb-
doscheelite Ca(W,Mo)O4, a representative of the isomorphous series of scheelite
CaWO4—powellite CaMoO4, and molybdenite. The deposit had been intensively developed starting from the middle of the XX
century. Flotation had always been the main concentration process used at the plant. When the global market for tungsten changed at the end of the XX century, the
Tyrnyauz concentrator was shutdown. By that time, the Main Skarn with tungsten and
molybdenum mineralization and a significant part of the amphibole-biotite hornfelses
with molybdenum mineralization had been mined out. At present, an upgrade of the
Tyrnyauz concentrator is required. After the enterprise is repeatedly put into operation,
its ore base will be represented by high-calcite skarn ore bodies (up to 10–15%) and © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 152–156, 2019. https://doi.org/10.1007/978-3-030-22974-0_36 Process Mineralogy as a Basis of Molybdoscheelite Ore Preparation 153 skarned marbles with the calcite content of 40–70%. It is known that calcite and
molybdoscheelite have similar flotation properties (Barskyi et al. 1979). Due to the high calcite to molybdoscheelite ratios, their selective flotation with the
use of traditional reagent regimes is impractical. Moreover, at lower mining horizons,
scheelite is almost completely replaced by molybdoscheelite, its mineral variety with
greater brittleness and lower hardness. These circumstances call for a new miner-
alogical and technological assessment of the Tyrnyauz deposit. Another independent
promising type of ores are talcose hornfelses with high molybdenite contents and
associated gold mineralization. However, the availability of a pair of naturally
hydrophobic minerals (molybdenite and talc) also hinders their flotation concentration
(Tarasevitch et al. 2014). 2
Methods and Approaches In both cases, the skarns are represented by thin lens-shaped and clustered
units (with the thickness of several centimeters to several tens of centimeters) having a
fine to medium-grained structure with the characteristic light greenish-brown color. Marbles in the skarn bodies are white, light gray and dark gray massive rocks with
a spotted, brecciated or banded texture caused by the alternating interlayers of dark and
light marbles of different granularity, alternating interlayers of light marbles and thin
banded light green pyroxene-plagioclase hornfelses and new units in the form of
wollastonite-fluorite,
pyroxene-fluorite
and
quartz-pyroxene-calcite-garnet
veins,
veinlets and lenses. The marbles are composed of calcite. The sizes of polysynthetic
calcite grains containing fine scattered inclusions of graphite and pyrrhotite range from
0.08 to 1.6 mm. In the skarned marbles, the distribution of structural, textural and color varieties of
marble is non-uniform. Within the contours of the ore body, fine, medium and non-
uniformly crystalline and, respectively, dark and light sulfur varieties predominate (in
quantitative terms). The lighter and white varieties (“clarified” marbles) form among
them a system of veinlets, lens-shaped interlayers, clusters and blocks and are part of
the parallel-banded varieties of marbles with calcite bands with a thickness of 1 to
5 cm. Their formation is associated with recrystallization along the fractures and
increased permeability zones, as well as in tectonic deformation processes. In addition,
lighter zones are always observed at the interfaces between marbles and skarns and
around quartz-silicate veins and veinlets. Luminescent properties of the ore minerals and host rocks of the Tyrnyauz deposit
were studied since it is possible to use these luminescent properties as indicators for
rapid diagnostics and as separation features in ore separation. Preliminary concentration of the Tyrnyauz ore, which may be implemented using
X-ray luminescent separation (RL), is required due to the processing of comparatively
low-grade ores with large amounts of diluting materials and high mineralization con-
trasts. The contrasts in the luminescent properties of minerals and, therefore, of rocks
and ores, may also be used for the rapid diagnostics of various natural and industrial
types of ores in the process of their transportation and sorting, for optimizing their
grinding conditions, as well as in their subsequent processing. The RL spectra were
registered in the continuous and staged scanning modes. In the continuous scanning
mode, with the constantly enabled source of excitation, the entire spectrum was
recorded. 2
Methods and Approaches Several hundred small and several dozens of large samples of promising ore types were
selected and analyzed. Additionally, during the period of active operation of the
deposit, the material composition and ore concentration indicators had been continu-
ously analyzed for the respective production processes. The grain-size distribution of molybdoscheelite (MSh) of different compositions
and powellite is due to the regular decrease in hardness and increase in brittleness with
the increase in the molybdenum content in the scheelite—powellite series. The higher
hydrophobicity of powellite and high-molybdenum MSh (with powellite inclusions),
which increases for particles of fine classes due greater specific surface values, pre-
determines their accumulation in the sulfide-molybdenum concentrate. In contrast, the
higher affinity between scheelite and fatty acids, as compared to MSh and powellite,
gives certain advantages in the process of rougher scheelite flotation and in respective
final treatment. The dependence of flotation properties of MSh in these operations on the com-
position, chemical and phase inhomogeneity of the particles enables the use of MSh
composition and dispersion values to evaluate the efficiency of flotation processes. Chemical sampling data on the contents of Mook and WO3, their ratio and related
dispersion and the results of MSh local composition analysis serve as the criteria for
assessing the chemical inhomogeneity of MSh. The color of luminescence, spectro-
scopic and kinetic parameters of excitation and radiation represent rapid test indicators
for MSh. The color of luminescence is effective when assessing the content of pure
scheelite with high hardness and flotability (blue luminescence) and of all varieties of
MSh with powellite characterized by low hardness and poor flotability (yellow lumi-
nescence). When predicting the processing properties of MSh, dispersion assessment
data is of greater significance. Therefore, the criteria used for identifying the processing
varieties of ores with account of the composition of MSh should include variations in
the composition of pure scheelite, molybdoscheelite and powellite. Similar distribution
of molybdenite oxidation values in the ore biotite hornfelses (with almost zero MSh L. Vaisberg et al. 154 content) occurring in parallel to the Slepaya deposit may be used to assess the content
and distribution of powellite. Skarns of the Slepaya deposit do not form independent bodies. These either form
part of the metasomatically altered skarned hornfelses, layered and massive marbles, or
are represented by relics in the areas of development of later metasomatic mineral
associations. Process Mineralogy as a Basis of Molybdoscheelite Ore Preparation Therefore, reproducible and comparable results in the continuous scanning mode
may only be obtained when the samples are irradiated to the saturation state, i.e. for
relatively long periods (tens of minutes or more, depending on the mineral). In this
connection, a method was developed for expressly obtaining the required information
on the spectra by means of high-speed analysis in pulsed mode with staged scanning of
the spectrum. Spectral and kinetic RL characteristics were studied for the following
minerals: scheelite and molybdenite, calcite, wollastonite, quartz and plagioclase. The
resulting RL spectra were interpreted by comparison of the data obtained with the
reference data. For all minerals, respective types of luminescence centers were deter-
mined and the issues of variability in luminescent properties were considered. For the
spectra obtained, the intensities, positions of the maximum and half-widths of the
intrinsic luminescence band associated with the [WO4] and [MoO4] groups were
established, as well as the wavelength and the intensity of the luminescence lines for
impurity ions of rare-earth elements isomorphically replacing calcium ions in the
scheelite structure. 2
Methods and Approaches This is the traditional and most informative form of data recording. However,
under the influence of X-rays, the luminescence intensity of a substance usually
increases. After a certain period of time that is specific for respective centers of
luminescence, the saturation level is reached, when the luminescence intensity prac-
tically ceases to change or changes so slowly that it produces no effect on the spectrum
image at any moment of the scanning time. 155 Process Mineralogy as a Basis of Molybdoscheelite Ore Preparation 3
Results and Discussion Visual observation of the luminescence for the molybdoscheelite samples studied
showed that many of them are characterized by a non-uniform color glow (blue and
yellow), which indicates joint availability of several varieties of scheelite. In this case,
the experimentally studied RL spectrum is composed of individual spectra of these
varieties and represents an averaged characteristic for the scheelites of the sample. The
scheelite and molybdoscheelite samples studied are characterized by varying degrees of
chemical composition inhomogeneity, which is manifested in different colors of their
fluorescent luminescence. The half-width of the band of intrinsic luminescence may be
used as the inhomogeneity measure. The observed diversity of the spectra may be explained by the presence of different
types of MSh in different sample groups with varying content ratios and by the pres-
ence of their spectra. Based on the spectroscopic data obtained, at least four varieties
with different luminescent properties may be distinguished for the scheelite samples
studied. Signs of vertical zoning are observed in the distribution of MSh with different
types of luminescence spectra. At the uppermost horizons, the luminescence intensity is
usually higher. The luminescence and afterglow spectra for all calcite samples studied consist of a
single broad band with the luminescence maximum of 630 nm (orange-red). It was also
found that luminescence intensity is variable and depends on a number of parameters,
such as the duration of x-ray irradiation, the concentration of manganese, and the color
of the samples. An important element of the promising processing technology for the
Tyrnyauz ores is the use of selective vibration in disintegration and classification of raw
materials. 156 L. Vaisberg et al. 4
Conclusions These studies in the field of process mineralogy of molybdoscheelite ores of the
Tyrnyauz deposit form the basis for the design of a new combined process flow using
various selective vibration methods for disintegration and preliminary lump separation. Acknowledgements. The study was supported by the grant of the Russian Science Foundation
(project No. 17-79-30056). Barskyi LA, Kononov OV, Ratmirova LI (1979) Selective flotation of calcium minerals. Nedra,
Moscow
Tarasevich Y, Aksenenko EV (2014) The hydrophobicity of the basal surface of talc. Colloid J
76(4):526–532 © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 157–161, 2019.
https://doi.org/10.1007/978-3-030-22974-0_37 1
Introduction The task of screening ore occurrences has always been and remains one of the
important tasks of the geological survey. One of the achievements of applied miner-
alogy in the history of its development is participation in metallogenic researches and
the development of the theory of mineral deposits prediction. Among the tasks solved
by the methods of applied mineralogy (in particular the search for crystallomorphol-
ogy) there are tasks aimed at determining the scale of mineralization (its vertical extent
and the magnitude of the erosion-denudation slice). The methods of the search crystal
morphology are based on two important postulates: the ontogenic development of the
mineral form and the doctrine of typomorphism. One of such methods is crystallo-
morphological method for estimating of tin ore occurrences developed by N. Z. Evzikova on the example of tin deposits. References arskyi LA, Kononov OV, Ratmirova LI (1979) Selective flotation of calcium minerals. Nedra
Moscow arasevich Y, Aksenenko EV (2014) The hydrophobicity of the basal surface of talc. Colloid
76(4):526–532 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Nizhny Novgorod Institute of Education Development,
Nizhny Novgorod, Russia
viann@inbox.ru Nizhny Novgorod Institute of Education Development,
Nizhny Novgorod, Russia
viann@inbox.ru Abstract. The methods of applied mineralogy are used to assess the scale of
the mineralization of tin occurrences and deposits. One of such methods is the
crystallomorphological method for estimating the level of tin ore bodies
developed by N.Z. Evzikova. According to the crystallomorphological features
of cassiterite it allows one to estimate the possible depth of distribution of
mineralization and its magnitude. Using the example of three studied tin-ore
districts of the Far East the possibilities of using crystallomorphology of cas-
siterite to predict and evaluate the prospect of mineralization of both the ore
region as a whole and individual ore occurrence at the exploration stage are
shown. Keywords: Applied mineralogy Crystallomorphology
Crystallomorphological method Cassiterite Scale of mineralization
An assessment of prospects Keywords: Applied mineralogy Crystallomorphology
Crystallomorphological method Cassiterite Scale of mineralization
An assessment of prospects Crystallomorphology of Cassiterite
and Its Practical Importance I. Vdovina(&) 2
Methods and Approaches Crystallomorphological method for evaluating of tin ore occurrences (Evzikova 1984)
is based on the difference in the natural appearance of cassiterite crystals in different
parts of the ore-bearing system as they move away from the source. The deeper parts of I. Vdovina 158 the ore bodies correspond to short-prismatic, isometric forms of cassiterite (I and II
types), the higher—elongated forms of crystals (IV and V). The scale of the miner-
alization and the degree of its preservation reflect the crystallomorphological criteria:
scoring represented by the statistical predominance of types IV and V and values from
50 to 200; degree of elongation of crystals (object’s class) reflecting the completeness
of the crystallomorphological evolution; the homogeneity of the morphology of cas-
siterite in different height sections of the ore object. The scoring factor is calculated by
the formula B = [2 (V) + (IV)] −[2 (I) + (II)], where I, II, IV, V is the percentage of
crystal morphological types in the sample. The testing of this method at various times took place at tin ore deposits of the Far
East where for the first time this method was used to evaluate the prospects for tin ore
mineralization in exploration. The author has researched the crystallomorphological
features of cassiterite from the ore manifestations of three Far Eastern tin ore regions
(the Komsomolsk, Badzhal, Yam-Alin regions) in order to obtain a crystallomorpho-
logical estimate of the prospect of their mineralization. 3
Results and Discussion Ore districts are located within the Khingan-Okhotsk volcano-plutonic belt and the
same-name tin-bearing region. The Khingan-Okhotsk volcano-plutonic belt represents
a vast magmatic area of the northeast strike superimposed on a heterogeneous foun-
dation formed in the atmosphere of the transform continental margin. The Badzhal
region is located in the junction zone of ancient Archean-Proterozoic cratons and
Mesozoic accretionary folded belts and is characterized by the presence of a Cretaceous
volcano-tectonic formation in the central part of the granitoid batholith. Komsomolsk and Yam-Alin regions are located in accretionary folded systems
spatially coincides with the eponymous volcanic zone (Rodionov 2005). The Komsomolsk Ore Region. Several hundreds of ore mineralization zones, mainly
cassiterite-silicate formation, have been established within the Komsomolsk ore region. By the beginning of 90s of last century crystallomorphological researches covered
about 40 manifestations of tin mineralization in which a fairly representative number of
samples were analyzed. 25 ore manifestations of this number were evaluated at deep
horizons in the process of prospecting and evaluation (14) or exploration which made it
possible to determine the reliability of forecasts obtained earlier by the crystallomor-
phological method. When sorting out 25 ore occurrences by crystallomorphological method 11 min-
eralization zones received a positive forecast. Subsequent testing at the exploration
stage confirmed this forecast for 6 ore occurrences. In five positively evaluated by the
subsequent preliminary exploration the forecast was not confirmed. Among the ore occurrences that received a negative assessment on the depth 10
objects were studied. The convergence of the results in these cases turned out to be
higher. A negative forecast was confirmed by 8 occurrences. And only in two zones
contrary to expectations industrial ores have been established. 159 Crystallomorphology of Cassiterite and Its Practical Importance The Badzhal Ore Region. The crystallomorphological researching of cassiterite from
the ore occurrences of the Badzhal ore district began with the first explorations in the
middle of 70s of the last century (Vdovina 2005). In the process of research at the exploratory stage the cassiterite ore occurrences of
the “Badzhal axis” structure were studied. Crystallomorphological criteria indicated a
high level of slice and insignificant prospects for ore occurrences. No detailed work has
been done. The Pravourmiyskoe deposit was the most studied in details. The morphology of
cassiterite crystals of this deposit was researched at all stages of prospecting and
exploration (Vdovina 1987, 2008). 3
Results and Discussion At the first stage of work the crystallomorphological mapping was carried out on
surface mine workings. Evaluation of the mineralization’s prospect was as follows: – the level of the denudation slice is high enough; – the gradient of the vertical crystallomorphological variability is large; the deposit is
characterized by the proximity of the upper and lower boundaries of mineralization; – the gradient of the vertical crystallomorphological variability is large; the deposit is
characterized by the proximity of the upper and lower boundaries of mineralization;
– the estimated length of the mineralization to the depth is 200–220 m; y
p
y
pp
– the estimated length of the mineralization to the depth is 200–220 m; – the morphology of the crystals suggests a large length of mineralization to the depth
and the occurence of industrial ores in the middle part of the deposit. Material was obtained from the deep horizons of the deposit in the detailed
exploration. The results of the crystallomorphological analysis of cassiterite of the
Pravourmiyskoe deposit were confirmed by the results of exploration. The length of the
mineralization to a depth was no more than 250 m. The level of the denudation slice is
really high. In addition the results of cassiterite of the Pravourmiyskoye deposit’s
crystallomorphological features researching confirmed the regularity of the spatial and
temporal distribution of the morphological types of cassiterite. The same sequence of
the cassiterite crystals changing from type I to type V was established at the stockwork
type of the cassiterite-quartz formation. The deposit is currently being exploited. About a dozen have been studied within the Verkhnebadzhalsky ore cluster. Most
detailed and from the surface and from the depth Blizhnee deposit has been explored
(Vdovina 2011). The Blizhnee Deposit. Cassiterite is of a short-prismatic almost isometric habit, IV and
less than V type, the score is 92–130. According to the crystallomorphological features
the prospect of mineralization to a depth is small. Exploration data did not confirm the
depth of the deposit. The Talidzhak mine is the most perspective one. Cassiterite is predominantly IV and
V types, of medium degree of elongation, the scaling varies within 100–200. The
gradient of crystal morphological variability is weak. Ore occurrences are characterized
as poorly eroded. The vertical extent of mineralization is assumed to be 500 m. The
detailed exploration has not been carried out. The Yam-Alin Ore Region. 3
Results and Discussion Ore occurrences were researched only at the exploration
stage. There is the Sorukan deposit. Cassiterite is situated in the zones of quartz-chlorite I. Vdovina 160 composition in the form of scattered and nested impregnation and quartz-cassiterite
lenses and veinlets. Pyramid-prismatic crystals of type IV of crystallomorphological
type prevail in their composition, crystals of type V are found in a small amount. There
are a lot of prismatic fragments of crystals of very different lengths including conical. All
type IV crystals are characterized by a moderate and strong degree of elongation, a zonal
structure; growth zones fix the development of cassiterite in types II–IV. The deposit’s
class development is 4. The deposit is poorly eroded and perspective on depth. The ore occurrences of the Shiroky, Exan and Bastion sections from the different
ones are of interest. Cassiterite is very small, almost colorless, slightly yellowish, often
observed in star splices and aggregates. It is presented by IV and V crystallomor-
phological types with a predominance of type V. The crystals are characterized by short
heads (dull pyramid), moderate and strong degree of elongation. The crystals are
transparent, zonal; the observed growth phantoms show their development in type IV
and III. Often there are simple articulated twins. The presence of cassiterite of different
physical and crystallomorphological properties gives the right to assume the devel-
opment of tin mineralization in several stages. In general the district is prospective,
poorly eroded. According to the results of the crystallomorphological method the most promising
sections include the majority of the studied sections. 4
Conclusions The author believes that the use of crystallomorphological analysis to determine the
denudation slice, the extent of mineralization to the depth and generally for the
industrial evaluation of tin mineralization is appropriate at the exploration stage. But
the credibility and reliability of the assessment will increase if it is used in combination
with others, in particular, at the first stage with structural and morphostructural
(Vdovina 2008), and at later ones with more “direct” mineralogical, geochemical and
other researches. Vdovina IA (1987) Crystallomorphology of cassiterite as one of the criteria for industrial
evaluation of the tin ore deposit in notes all-union. Mineral Soc Part XVI, I:60–65
Vd
i
IA (2005) C
ll
h l
i
l f
f
i
i
i
h B d h l Evzikova NZ (1984) Prospecting cristal morphology, Moscow
Rodionov SM (2005) Metallogeny of tin of East of Russia, Moscow
Vdovina IA (1987) Crystallomorphology of cassiterite as one of the criteria for industrial
evaluation of the tin ore deposit in notes all-union. Mineral Soc Part XVI, I:60–65
Vdovina IA (2005) Crystallomorphological features of cassiterite occurrences in the Badzhal ore
region. Nat Geogr Res, Komsomolsk-na-Amure, pp 4–13
Vdovina IA (2008) Morphostructural-crystallomorphological evaluation of the prospect of
mineralization (on the example of the Badzhal tin ore region) in Fedorov session 2008.
Thesises of international scientific conferences, Saint-Petersburg, pp 246–249 Vdovina IA (2011) About the crystallomorphology of cassiterite. In: X international conference
«New ideas in the Earth sciences». Reports in 3 volume. Russian State Geological
Exploration University, Moscow, vol 1, pp 107–108 Crystallomorphology of Cassiterite and Its Practical Importance References Evzikova NZ (1984) Prospecting cristal morphology, Moscow Evzikova NZ (1984) Prospecting cristal morphology, Moscow Rodionov SM (2005) Metallogeny of tin of East of Russia, Moscow Vdovina IA (1987) Crystallomorphology of cassiterite as one of the criteria for industrial
evaluation of the tin ore deposit in notes all-union. Mineral Soc Part XVI, I:60–65 Vdovina IA (2005) Crystallomorphological features of cassiterite occurrences in the Badzhal ore
region. Nat Geogr Res, Komsomolsk-na-Amure, pp 4–13 Vdovina IA (2005) Crystallomorphological features of cassiterite occurrences in the Badzhal ore
region. Nat Geogr Res, Komsomolsk-na-Amure, pp 4–13 Vdovina IA (2005) Crystallomorphological features of cassiterit
region. Nat Geogr Res, Komsomolsk-na-Amure, pp 4–13 Vdovina IA (2008) Morphostructural-crystallomorphological evaluation of the prospect of
mineralization (on the example of the Badzhal tin ore region) in Fedorov session 2008. Thesises of international scientific conferences, Saint-Petersburg, pp 246–249 Vdovina IA (2008) Morphostructural-crystallomorphological evaluation of the prospect of
mineralization (on the example of the Badzhal tin ore region) in Fedorov session 2008. Thesises of international scientific conferences, Saint-Petersburg, pp 246–249 Crystallomorphology of Cassiterite and Its Practical Importance 161 Vdovina IA (2011) About the crystallomorphology of cassiterite. In: X international conference
«New ideas in the Earth sciences». Reports in 3 volume. Russian State Geological
Exploration University, Moscow, vol 1, pp 107–108 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction Modal (quantitative mineralogical) analysis of rocks and ores in thin sections is one of
the first fundamental quantitative methods of mineralogy (including technological
mineralogy), petrography (in classifications of rocks and ores, and petrological
reconstructions), and lithology (i.e., petrography of sedimentary rocks). That is why its
rigorous justification is of fundamental importance owing to the Delesse-Rosiwal-
Glagolev ratios, as well as to stereological reconstruction. It makes sense to summarize
the history of these methods in Russia and abroad, and to formulate the conclusion
about their prospects. Yu. Voytekhovsky(&) Yu. Voytekhovsky(&) Saint-Petersburg Mining University, Saint-Petersburg, Russia
Voytekhovskiy_YuL@pers.spmi.ru Saint-Petersburg Mining University, Saint-Petersburg, Russia
Voytekhovskiy_YuL@pers.spmi.ru Saint-Petersburg Mining University, Saint-Petersburg, Russia
Voytekhovskiy_YuL@pers.spmi.ru Abstract. The article is devoted to the history and justification of the modal
analysis of rocks and ores with a microscope. It is shown that the Delesse-
Rosiwal-Glagolev ratios do not follow from the Cavalieri principle. They do not
allow one to find the exact volume of the minerals in rocks or ores, but give only
their average estimates. It is also shown that the volume fractions of convex
mineral grains in rocks and ores, taken equal to the fractions of their flat sec-
tions, are always underestimated if compared with the matrix. Due to the wide
variety and complexity of forms of mineral grains, the methods of stereological
reconstruction lead to integral equations with a difficult to define form factor. Most likely, tomography methods should come to replace the modal analysis of
rocks and ores in thin sections. Keywords: Rocks and ores Modal analysis Stereological reconstruction 2
The Cavalieri Principle and the Delesse-Rosiwal-Glagolev
Ratios First of all, we point out that the relations suggested by Delesse (1848) dVi = dSi,
Rosiwal (1898) dSi = dLi, and Glagolev (1932) dLi = dNi, decreasing the dimension of
space (namely, equating the volume fractions of minerals to areal, areal to linear, linear
to point-like), have no relation to the Cavalieri principle: S1i = S2i ! V1 = V2 (if the
areas of all arbitrarily close parallel sections of two bodies are pairwise equal, then their
volumes are also equal, Fig. 1). © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 162–166, 2019. https://doi.org/10.1007/978-3-030-22974-0_38 Modal Analysis of Rocks and Ores in Thin Sections 163 Fig. 1. To the justification of the Cavalieri principle Fig. 1. To the justification of the Cavalieri principle In recent notation, this principle, historically preceding integral calculus, has a clear
meaning: V1 ¼
R
S z
ð Þ dz ¼ V2, where S(z) is a continuous function of the areal
fraction of a certain mineral along the z axis being normal to the sections. But, the
modal analysis of rocks and ores in thin sections, which accumulates the statistics of
areal, linear or point fractions of minerals from section to section, has nothing to do
with the integration procedure. It only leads to an assessment of their average values. In
this case, it can be argued that the volume of any mineral in a rock or ore is within the
certain interval: S z
ð ÞminDz ¼
Z
S z
ð Þmindz\V\
Z
S z
ð Þmaxdz ¼ S z
ð ÞmaxDz where Dz is the thickness of rock sample under study. Despite of this contradiction, which was also considered in the works (Krumbein
1935; Chayes 1956), the method became firmly established in practice because of its
apparent simplicity and was step by step automated (Shand 1916; Wentworth 1923;
Hunt 1924; Dollar 1937; Hurlbut 1939) up to the use of modern computers for image
analysis of thin sections. The list of parameters characterizing the cross-section of
minerals, and the speed of processing have grown many times. But, in terms of the
reconstruction of the true metric characteristics of mineral grains from those of their flat
or even linear sections, the ideology remains the same. Companies that produce computer structure analyzers offer software packages
without discussing the fundamental problems. The analysis of 2D images does not use
the available chapters of mathematics. 3
Stereological Reconstruction A new line of research, i.e. stereological reconstruction, arose from the obvious
observation that an arbitrary cross section of a spherical shape is always less than its
characteristic cross section (Fig. 2, above). And it follows from this that the volume
fraction of the convex mineral phase in the rock and ore, equal to the fraction of its flat
sections, is always underestimated if compared with the host matrix. The corresponding
general problem – finding the distribution of true particle sizes from the size distri-
bution of their random sections – belongs to the inverse problems typical for geo-
physics. It is analytically solved only for spherical and ellipsoidal particles due to
relatively simple description of these forms (Wicksel 1925, 1926). But, even in this
case, the practical use of the theory requires the selection of the best solution and an
estimate of the errors (Fig. 2, below). For more complex forms of mineral grains this
can’t be done without mathematical modeling on powerful computers. Fig. 2. Left: the size of the cross section of a convex grain is always less than the characteristic
one, in the modal analysis its volume fraction is underestimated. Right: to recognize the true size
of spherical particles by the size of circular sections; R = 100 – particles with a radius of 100
arbitrary units; R = 100, r = 50 – two sets of particles of the same type, indistinguishable in
sections (for example, two generations of one mineral); horizontal scale – section radii from 0 to
100 bits per 10 classes; vertical scale – frequencies by classes (lower curves) and accumulated
frequencies (upper curves). Fig. 2. Left: the size of the cross section of a convex grain is always less than the characteristic
one, in the modal analysis its volume fraction is underestimated. Right: to recognize the true size
of spherical particles by the size of circular sections; R = 100 – particles with a radius of 100
arbitrary units; R = 100, r = 50 – two sets of particles of the same type, indistinguishable in
sections (for example, two generations of one mineral); horizontal scale – section radii from 0 to
100 bits per 10 classes; vertical scale – frequencies by classes (lower curves) and accumulated
frequencies (upper curves). The history of this area in Russia can be found in the following incomplete list of
works: Zhuravsky A.M. 2
The Cavalieri Principle and the Delesse-Rosiwal-Glagolev
Ratios For example, the distances between the mineral
grains in thin section are replaced by the Euclidean distances between the points taken
within the grains, whereas there is a more complicated, but easily programmable
Hausdorff’s metric which allows one to do this procedure correctly. In turn, it gives us
the possibility of calculating space covariograms between the mineral grains of dif-
ferent species and their various clusters in the rock. Yu. Voytekhovsky 164 4
Conclusions Thus, due to the extraordinary diversity and complexity of the forms of mineral grains
in rocks and ores, the methods of stereological reconstruction lead to integral equations
with an analytically difficult-to-define form factor. The practical application of the
theory is drowning in the selection of the best solution to the inverse problem and
complex estimates of measurement errors. It seems that the modal analysis of rocks and
ores in thin sections should be replaced by tomography methods. Standardizing modal
analysis of rocks and ores in thin sections by creating their artificial counterparts with
previously known volume fractions of mineral grains and a wide range of petrographic
structures can serve as an inter-laboratory comparison of the accuracy of the method. But it does not solve the problems in essence. 3
Stereological Reconstruction Mineralogical analysis of thin section in terms of probabilities. Moscow-Leningrad: Gosgeolizdat, 1932. 20 p.; Glagolev A.A. Quantitative miner-
alogical analysis of rocks with the microscope. Leningrad: Gosgeolizdat, 1932. 25 p.;
Glagolev A.A. On the geometric methods of quantitative mineralogical analysis of
rocks. Moscow-Leningrad: Gosgeolizdat, 1933. 47 p.; Glagolev A.A. Geometric
methods for quantitative analysis of aggregates with a microscope. Moscow-Leningrad:
Gosgeolizdat, 1941. 263 p.; Chayes F. An elementary statistical appraisal. New York:
John Wiley & Sons, Inc., 1956; Shvanov V.N., Markov A.B. Granulometric analysis of
sandstones in thin sections//Geology and exploration. 1960. N 12. P. 49–55; Ivanov N. V. A new direction in testing ore deposits. Moscow: Gosgeolizdat, 1963. 179 p.;
Chernyavsky K.S. Stereology in metallurgy. Moscow: Metallurgy, 1977. 375 p.;
Ivanov O.P., Ermakov S.F., Kuznetsova V.N. Improving the accuracy of determining
the weight particle size distribution of ore minerals from measurements in thin 165 Modal Analysis of Rocks and Ores in Thin Sections sections//Proc. CNII of Tin. Novosibirsk: Science, 1979. p. 10–14; Gulbin Yu.L. On
stereological reconstructions of grain sizes in aggregates//Proc. Rus. Miner. Soc. 2004. N 4. P. 71–91. sections//Proc. CNII of Tin. Novosibirsk: Science, 1979. p. 10–14; Gulbin Yu.L. On
stereological reconstructions of grain sizes in aggregates//Proc. Rus. Miner. Soc. 2004. N 4. P. 71–91. Wicksel SD (1926) The corpuscle problem: case of ellipsoidal corpuscles. Biometrica 18:151 Wicksel SD (1926) The corpuscle problem: case of ellipsoidal corpuscles. Biometrica 18:151
172 Quality Assurance Support (QA/QC System)
of Mineralogical Analysis O. Yakushina(&), E. Gorbatova, E. Ozhogina, and A. Rogozhin FSBE “All-Russian Institute on Mineral Raw Materials” (VIMS),
Moscow, Russia
vims@df.ru Abstract. Mineralogical studies are an integral part of the exploration and
development of solid mineral deposits, the effectiveness of which directly
depends on the quality of the measurements. Moreover, five decades QA/QC
system of mineralogical analysis (UKARM) for Russian geological survey is
developed. Its specific features and tasks are discussed. The system provides to
obtain complete, reliable, metrologically evaluated and legally valid information
about the material composition and structure of rocks and ores. QA/QC of
mineralogical analysis ensures the coordination of testing laboratories, starting
with the resources of testing laboratories, stuffing at last. This system covers the
entire process of mineralogical research, starting at the resources of testing
laboratories as stuff and equipment, through the research procedure as the
selection of testing object and its preparation for analysis, the accuracy of
analysis rank, the testing method and technique, to metrology data and the
quality assessment of the results obtained. Keywords: Mineralogical analysis Quality management Assurance
Control system Metrology Reference materials References Delesse M (1848) Procede mecanique pour determiner la composition des roches. In: annales des
mines. De memoires sur l’exploitation des mines. 4me serie. T. XIII. Carilian-Goeury et
Dalmont, Paris, pp 379–388 Dollar ATJ (1937) An integrating micrometer for the geometrical analysis of rocks. Mineral Mag
24:577–594 Hunt WF (1924) An improved Wentworth recording micrometer. Am Mineral 9:190–193
Hurlbut CS Jr (1939) An electric counter for thin-section analysis. Am J Sci 237:253–261 Hunt WF (1924) An improved Wentworth recording micrometer. Am Mineral 9:190–193
H rlb t CS Jr (1939) An electric co nter for thin section anal sis Am J Sci 237:253 261 Krumbein WC (1935) Thin-section mechanical analysis of indurated sediments. J Geol 43:482–
496 Rosiwal A (1898) Über geometrische Gesteinanalysen. Ein einfacher Weg zur ziffermässigen
Feststellung des Quantitätsverhältnisses der Mineralbestandtheile gemengter Gesteine. In:
Verh. der k.-k. Geol. Reichsanstalt. Verlag der k.-k. Geol. Reichsanstalt, Wien, pp 143–175 g
pp
nd SJ (1916) A recording micrometer for geometrical rock analysis. J Geol 24:394–404 Wentworth CK (1923) An improved recording micrometer for rock analysis. J Geol 31:228–232
Wicksel SD (1925) The corpuscle problem: a mathematical study of a biometric problem. Biometrica 17:84–99 Wentworth CK (1923) An improved recording micrometer for rock analysis. J Geol 31:228–232 Wicksel SD (1925) The corpuscle problem: a mathematical study of a biometric problem. Biometrica 17:84–99 Wicksel SD (1926) The corpuscle problem: case of ellipsoidal corpuscles. Biometrica 18:151–
172 172 Yu. Voytekhovsky 166 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction Mineralogical analysis is an integral part of mineral deposits exploration and devel-
opment; its effectiveness directly depends on the quality of testing (ISO/IEC
17025:2005; JORC Code 2012). The quality of analysis directly affects efficiency
and reliability of the whole investigation. The main requirement for mineralogical
research is to obtain reliable, complete, metrologically evaluated and legally valid
information about the studied matter composition and structure, namely of rocks, ores
and technologically processed products. The quality management system of miner-
alogical works ensures the coordination of testing laboratories, starting with the
resources of testing laboratories, and ending with the processes occurring in them. Laboratory mineralogical studies with varying degrees of depth of mineral substance
analysis are carried out at all stages of geological exploration from geological
prospecting to operational exploration and development of mineral deposits. The main
requirement for laboratory mineralogical studies is to provide all spheres of activity of
the Ministry of Natural Resources and Ecology of the Russian Federation with reliable, © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 167–171, 2019. https://doi.org/10.1007/978-3-030-22974-0_39 O. Yakushina et al. 168 reliable, standardized, metrologically evaluated and legally valid information obtained
as a result of the use of a wide range of laboratory methods and equipment (Ozhogina
et al. 2017a, b). reliable, standardized, metrologically evaluated and legally valid information obtained
as a result of the use of a wide range of laboratory methods and equipment (Ozhogina
et al. 2017a, b). 2
Methods The variety and complexity of objects of natural and man-made origin, the widespread
use of quantitative methods of mineralogical analysis, the presence of a large number of
mineralogical laboratories with different instrumentation and methodological base,
personnel composition, determines the need to improve and develop the Quality
Management System QA/QC for Mineralogical works UKARM, established at VIMS
in the 1980th (Ginsburg et al. 1985) and renovated today. Developed by VIMS the
UKARM System coordinates laboratory studies, preparation for testing operations and
also monitors the quality of research. Scientific-methodical support of the UKARM
functioning carries out the Scientific Council on methods of mineralogical methods of
analysis (NSOMMI). The main components of UKARM mineralogical QA/QC system
include as follows: – make the General concept of development of mineralogical service and deliver
priority directions of its improvement; – preparation of proposals for the program and coordination plan of scientific research
aimed at solving mineralogical laboratory research methods of testing; – develop the industry system of standard samples, coordinate their production,
registration and use; – the NSOMMI Scientific Council review, update and elaborate new guidance doc-
uments on methods of all kinds of laboratory mineralogical works, the system and
means of these documents verification and approval; – ensuring the functioning of a unified quality management system of laboratory
mineralogical work, organization and carrying the interlaboratory comparative tests
(ICT), internal and external laboratory analysis control; – methodological assistance support for the basic and regional laboratories and
research centers in certification of mineralogical methods of analysis; – implementation of complex and unique mineralogical and analytical studies; – development of operational information system for mineralogical research. UKARM QA/QC system is based on the requirements of the requirements of
ISO/IEC 17025 standard in relation to mineralogical research, starting with the
resources as stuff and equipment, ending with metrology and data quality assessment
(Fig. 1). The testing laboratory stuff must have the necessary competence to perform at
a high level of mineralogical works. UKARM QA/QC includes workshops, advanced
training courses for laboratory stuff. For example, every year, since 2011, VIMS
arrange annual Seminar “Mineralogical school – Current problems and modern
methods”. 2
Methods Seminar members discuss the basic state-of-art concepts of mineralogy,
prospects of development, mineralogical support of geological exploration, methods of
mineralogical analysis, environmental problems, specific features of man-made raw
materials study, mineralogy for enrichment of ores, the nature of minerals technological Quality Assurance Support (QA/QC System) of Mineralogical Analysis 169 properties and their behavior in geological and technological circulation, mineralogical
works metrological and methodological support, etc. Specialists of various affiliation
took part in the Seminar. Fig. 1. UKARM QA/QC system structure Fig. 1. UKARM QA/QC system structure Testing laboratories are equipped with the necessary units: measuring instruments,
software, standards, reference data, reagents and consumables. Also the methodical
base is developed or adapted to technical base of analysis current state, corresponding
metrological assessment and its legal approve. The certification and registration of
standard samples of phase composition and properties, and artificial mixtures is also a
necessary element of QA/QC. The choice of mineral substance testing method and
mode is determined by substance peculiarities in composition and structure, the
hardware capabilities, the availability of relevant methodological documents and the
staff competence. In 2018 the Register on mineralogical studies of natural or techno-
geneous mineral raw matter includes 3 industry standards, 45 instructions, 31 practical
guidelines and 174 recommendations. Today, the methods of quantitative phase analysis of rocks, ores and man-made
substance are especially in demand. UKARM QA/QC includes the classification of
quantitative phase analysis laboratory methods, depending on their reproducibility, are
divided into six categories: I – particularly accurate quantitative analysis, II – full phase
analysis with increased accuracy, III – ordinary quantitative analysis, IV – ordinary
analysis with reduced accuracy requirements (express), V – semi-quantitative analysis,
VI – qualitative phase analysis. Any testing may/should be characterized by a level of
accuracy (Ozhogina et al. 2017a, b). 4
Conclusions Mineralogical study of any substance should be based on a QA/QC System. Testing
laboratory, which performs mineralogical studies supporting geological exploration
and technological works in order to meet the requirements of mineralogical work
quality management should have the relevant stuff, modern test methods, proper
equipment, capabilities and means of verification and calibration, be supplied by
industry techniques procedures, guidelines, as well as strictly follow them, and comply
with the requirements of a unified quality control system. 3
Results and Discussion The quality of measurements is provided by measuring units’ verification and cali-
bration tests; the availability of measurement guidance and their strict observance;
internal and external laboratory control. Organization of the control and dispatching
service for mineralogical analysis and interlaboratory comparative tests, carrying out
the control of phase composition and properties by standard samples, certified O. Yakushina et al. 170 mixtures; participation in analysis to certify the standard samples of phase composition
and properties, certification of laboratories with mandatory experimental evaluation. We state the leading role and the one of the most effective forms of external control
are Interlaboratory Comparative testing (ICT). The last one allow to evaluate the
reliability of the results obtained in each individual laboratory, and to obtain infor-
mation about the real accuracy of measurement techniques in general. In 2016–2018
VIMS conducted a series of ICT on X-ray powder diffraction (XPD) and petrography
analysis. The samples for control were artificial mixture of minerals (2), and synthetic
corundum powder (1), igneous (1) and sedimentary (1) rocks thing-sections. Each ICT
involved from 8 to 12 participants, totally 46 laboratories. The obtained results are
considered satisfactory, they can be used by laboratories during accreditation proce-
dures and confirmation of technical competence. The ICT comparative testing provide
an opportunity to assess the quality of measurements in different laboratories, to carry
out corrective actions to ensure the uniformity of measurements and to show the
technical competence of the laboratory. Mineral Preparation in Geological Research
T. Yusupov(&), A. Travin, S. Novikova, and D. Yudin
V.S. Sobolev Institute of Geology and Mineralogy SB RAS, Novosibirsk, Russia
yusupov@igm.nsc.ru V.S. Sobolev Institute of Geology and Mineralogy SB RAS, Novosibirsk, Russia
yusupov@igm.nsc.ru Abstract. Paper deals with discussion of minerals’ preparation requiring high
purity monoproducts, this has special importance for minerals –geochronome-
ters. Widely used methods including gravitation, magnetic separation, floatation
provide fractions with 90% of targeted mineral. Further monominerality increase
requires special separation methods; one of them – “Strat” is perspective. It is
based on separation in organic liquids under gradual density change. Combi-
nation of bromoform with d – 2.89 g/sm3 and dimethyle formamide with d –
0.8 g/sm3 is used; density gradations till 0.001 g/sm3 are possible therefore
isomorphic inclusions could be separated. Another direction is presented by
trybotreatments under higher energies in planetary mills with a centrifugal factor
to 40–50 g. Exotic surface substances presented mainly by kaolinite, muscovite,
calcite, gothite are removed as trybotreatment result. Special planetary mills -
classifiers are used for processing of big samples. This method together with
minerals opening in disintegrator under destruction by the free pulse is rec-
ommended for wide application. Keywords: Monominerality Geo chronometers Separation Surface
Organic liquids Trybotreatment Keywords: Monominerality Geo chronometers Separation Surface
Organic liquids Trybotreatment © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 172–175, 2019.
https://doi.org/10.1007/978-3-030-22974-0_40 References Ginsburg AI, Vikulova LP, Sidorenko GA (1985) On some typical mistakes at mineralogical
investigation (studies) J. In: Zapiski RMO (Proceedings of RMS), vol 3, pp 324–333 JORC Code: 2012 Edition. htt p
j
g
ISO/IEC 17025:2005: General requirements for the competence of testing and calibration
laboratories Ozhogina EG, Gorbatova EA (2017a) System of mineralogical quality management. J Actual
Probl Min 1:3–7 Ozhogina EG, Lebedeva MI, Gorbatova EA (2017b) Interlaboratory comparison tests in the
mineralogical works. J. Stand. Samples 2:37–47 171 Quality Assurance Support (QA/QC System) of Mineralogical Analysis Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Mineral Preparation in Geological Research
T. Yusupov(&), A. Travin, S. Novikova, and D. Yudin
V.S. Sobolev Institute of Geology and Mineralogy SB RAS, Novosibirsk, Russia
yusupov@igm.nsc.ru 2
Methods, Approaches, Results and Discussion We develop new section in sample preparation – mineral preparation, which includes
number of new research and methodological aspects. 1. Preliminary concentrating of minerals – chronometers from objects with very low
their content. Only intermediate products which are used for mono mineral fractions
extraction could be obtained by traditional types of separation – gravity, magnet and
floating. 1. Preliminary concentrating of minerals – chronometers from objects with very low
their content. Only intermediate products which are used for mono mineral fractions
extraction could be obtained by traditional types of separation – gravity, magnet and
floating. Gravitation methods are helpfully used in the situations where differences between
density of extracted mineral and monaural basis are not lower than 3 kg per sm3. Losses of target mineral are significant under other combinations of densities. Magnet
methods are more effective when differences in magnet sensitivity of separating
components are sufficient for separation. This is true for the case of quartz from −0.40
to 0.10 and biotite from + 46.7 to 86.7 109 m3/g. Under lower differences in magnet
sensitivity of minerals which are contained in samples at the level of accessory units
extraction is extremely difficult. Great perspectives in the extraction of minerals from extremely poor subsurface
rocks are related with floatation process which makes it possible to get minerals with
less than 0.1% content in sample. Obtained products of preliminary concentration
should either be further grinded in order to open intergrowths or be dressed with the
help of special concentrating methods till mono mineral state. Combine schemes with
mono mineral and similar products extractions at the initial stage are often used; then
intergrowths minerals opening and repeated concentrating of targeted minerals take
place (Berger 1962). 2. Opening of minerals from intergrowth stage done with the help of mechanical
treatments may be accompanied by significant structural chemical changes. It is
necessary to avoid high temperatures, local high pressures and if possible to use dry
process. High energetic free pulse realized in desintegrators is effective method of
minerals – chronometers’ opening. Prospectivity of desintegrator’s use for ore
preparation is proved on the cases of different minerals: spodumene, apatite, sulfides
and others. Higher preservation of crystal structure and lower over grinding are
considered to be main advantages here (Yusupov et al. 2015). Positive aspects of
mineral preparation were revealed under disintegrating of quartz – feldspar asso-
ciations (Yusupov et al. 2018). 1
Introduction Many mineralogical and geochemical researches are based on mono mineral products’
studies; acquisition of these products is based on research intensive processes of
minerals’ revealing and extracting. This assumption is relevant to geochemical, geo-
physical, lithologic, petrochemical and other studies; geochronological definitions
became wider during recent years. The task to extract minerals – chronometers of mono mineral purity is very
important and complicated. The problem becomes much more complicated when
chronometers are extracted from geo objects which have precious and rare metal
character. This happens due to their very low content, thin dispersion and occurrence in
genetic association (intergrowth) with usually rock-forming minerals. Range of rock-forming and ore minerals which are used for rock dating is
widening. Analysis is concentrated on such minerals as plagioclases, olivine, ortho and
clino pyroxene, phlogopite, tourmaline, sphalerite, volframite, tin spar, pyrite, pyr-
rhotite, pentlandite and others. Special methods should be developed for the extraction
of many of these minerals (Isotope…, 2015; Methods…, 2018). © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 172–175, 2019. https://doi.org/10.1007/978-3-030-22974-0_40 Mineral Preparation in Geological Research 173 2
Methods, Approaches, Results and Discussion Disintegrated sample preparation is recommended
for wide use. 3. Obtained concentrates were dressed by methods of mono mineral fractions
extraction with extraction of products with 90% of targeted mineral (Methods…
1985). Further increase of mono mineral character is reached by the help of special
methods. Gradual separation in organic liquids on density – “Starts” method and
trybo treatment – surface attrition under higher energies of mechanical treatment are
wide used. Mixture of bromform with d – 2.89 g/sm3 and dimethyle formamide
with d – 0.8 g/sm3 is used as separation media. Different density gradations till
0.001 g/sm3 are possible here. Potential of the method is shown on the example of
quartz – feldspar associations with – 0.3 +0.2 mm size (Table 1). 3. Obtained concentrates were dressed by methods of mono mineral fractions
extraction with extraction of products with 90% of targeted mineral (Methods…
1985). Further increase of mono mineral character is reached by the help of special
methods. Gradual separation in organic liquids on density – “Starts” method and
trybo treatment – surface attrition under higher energies of mechanical treatment are
wide used. Mixture of bromform with d – 2.89 g/sm3 and dimethyle formamide
with d – 0.8 g/sm3 is used as separation media. Different density gradations till
0.001 g/sm3 are possible here. Potential of the method is shown on the example of
quartz – feldspar associations with – 0.3 +0.2 mm size (Table 1). 174 T. Yusupov et al. Table 1. Reparability of minerals of non electromagnetic quartz feldspar product
Fraction density g/sm3 Output, % Elements content, %
SiO2 Fe2O3 Al2O3 K2O Na2O Li2O
Initial
81.1
0.03
12.3
2.48 5.18
0.028
2.44–2.55
7.881
65.3
0.02
19.3
13.3 1.28
0.097
2.55–2.58
3.751
2.58–2.61
5.109
2.61–2.63
43.795
71.0
0.03
18.4
0.51 9.82
0.004
2.63–2.65
35.737
98.7
0.03
1.4
0.12 0.62
0.017 Potassic feldspar product with 7.88% output was extracted under 2.44–2.55 g/sm3
density. Fraction of sodium feldspar is concentrated under higher density d – 2.61–
2.63 g/sm3 with output of 43.79%. Density of quartz fraction is more close to similar
indicator for sodium spar with density interval being 2.63–2.65 g/sm3 and output
35.73%. These results confirm high effectiveness of density method; separation of 0.05 mm
and lower size products looks possible (Yusupov et al. 2015). Method is successfully
used for not only quartz and feldspar separation but also for muscovite, biotite, glau-
conite (Katz 1977). Mineral Preparation in Geological Research Mineral Preparation in Geological Research 3
Conclusions Methodical bases of samples preparation in processes of mineral products extraction
are reviewed. Taking into account orientation of these methods it is suggested to name
this approach mineral preparation. Possibilities of minerals separation by “Strat” method which is based on use of
organic liquids with different density and surface trybo treatment are discussed. Mono
mineral products of high quality are obtained as a result of methods application. These methods as well as disintegrated minerals opening are recommended for
wide application in analytical practice. 2
Methods, Approaches, Results and Discussion Trybo attrition impacts are under investigated though they are important for min-
eral’s homogeneity increase. They enable to remove inclusions of tramp substances
presented mainly by kaolinite, muscovite, calcite, gothite as well as by remaining
floating reagents. It is important to take into account that after removal of surface layers of 0.1–10
mcm thickness surface is characterized by different structural imperfections. They
could vary from practically unchanged state to totally crystal and chemically destroyed
surface (Yusupov et al. 2018). Trybo treatment looks as important way to increase mono mineral character. Hand
attrition in jet and jasper stamps is widely used in institute’s analytical practice. Method
is applied for treatment of biotite, glauconite, amphibolites, tourmolin spinels, phos-
phates, sulphides and other minerals with monomimeral coefficients being about 100%. Facilities of PMK type are effective for large samples with weight more than 1 kg
trybo treatment. Material here is exposed by planetary rotating movement of ore mass. Such technological regime selectively destroys impurity substances and increases
monomineral properties of extracting products. Improved version of this mechanism is
being developed at CJSC “Itomak”, Novosibirsk. Trybotreatment not only removes impurity substances but also changes surface
defectiveness character and mineral heterogeneity type. Heterogeneity management is
under investigated methodologically however its role in ores processing is constantly
growing. For example concentrate with 0.05% ferrous oxide was got from quarts with
1.5% of this component. This result could not be reached by other methods. It is
important to provide transfer to homogeneous state when minerals have similar char-
acter of structural defects. Type and level of trybo treatment make it possible to solve
these problems to certain extent. 175 References Berger GS (1962) Minerals’ floatating. Gortechizdat, Moscow
Ginzburg AI (1985) Mineral research methods Nedra, Moscow g
(
)
g
,
Ginzburg AI (1985) Mineral research methods Nedra, Moscow g
Isotope dating of geological processes: new results, approaches and perspectives (2015) VI
Russian conference in isotope geochronology. IPGG RAS, Saint Petersburg Isotope dating of geological processes: new results, approaches and perspectives (2015) VI
R
i
f
i
i
t
h
l
IPGG RAS S i t P t
b Katz MY (1977) Minerals’ heterogeneity analysis. Nauka, Moscow Methods and geo chronological results of isotope geometric minerals systems and ores studies
(2018) VII Russian conference in isotope geochronology, Moscow Methods and geo chronological results of isotope geometric minerals s
(2018) VII Russian conference in isotope geochronology, Moscow Yusupov TS, Baksheeva II, Rostovtsev VI (2015) Analysis of different-type mechanical effects
on selectivity of mineral dissociation. J MiningSci 51:1248–1253 Yusupov TS, Baksheeva II, Rostovtsev VI (2015) Analysis of different-type mechanical effects
on selectivity of mineral dissociation. J MiningSci 51:1248–1253 Yusupov TS, Travin AV, Yudin DS, Novikova SA, Shumskaya LG, Kirillova EA (2018)
Improvement of methods of minerals – geo chronometers extraction from ores for geological
processes isotope dating. In: Shchiptsov VV (ed.) Fundamental and applied aspects of
technologica lmineralogy, Petrozavodsk. Russian conference in isotope geochronology,
pp 86–92 Yusupov TS, Travin AV, Yudin DS, Novikova SA, Shumskaya LG, Kirillova EA (2018)
Improvement of methods of minerals – geo chronometers extraction from ores for geological
processes isotope dating. In: Shchiptsov VV (ed.) Fundamental and applied aspects of
technologica lmineralogy, Petrozavodsk. Russian conference in isotope geochronology,
pp 86–92 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. V. Afanasiev1(&), N. Pokhilenko1, A. Eliseev1, S. Gromilov2,
S. Ugapieva3, and V. Senyut4 1 VS Sobolev Institute of Geology and Mineralogy, Siberian Branch,
Russian Academy of Sciences, Novosibirsk, Russia
avp-diamond@mail.ru
2 Nikolaev Institute of Inorganic Chemistry, Siberian Branch,
Russian Academy of Sciences, Novosibirsk, Russia
3 Diamond and Precious Metal Geology Institute, Siberian Branch,
Russian Academy of Sciences, Yakutsk, (Sakha) Yakutia, Russia
4 Joint Institute of Mechanical Engineering of the NAS of Belarus,
Minsk, Belarus 1 VS Sobolev Institute of Geology and Mineralogy, Siberian Branch,
Russian Academy of Sciences, Novosibirsk, Russia
avp-diamond@mail.ru
2 Nikolaev Institute of Inorganic Chemistry, Siberian Branch,
Russian Academy of Sciences, Novosibirsk, Russia
3 Diamond and Precious Metal Geology Institute, Siberian Branch,
Russian Academy of Sciences, Yakutsk, (Sakha) Yakutia, Russia
4 Joint Institute of Mechanical Engineering of the NAS of Belarus,
Minsk, Belarus 1 VS Sobolev Institute of Geology and Mineralogy, Siberian Branch,
Russian Academy of Sciences, Novosibirsk, Russia
avp-diamond@mail.ru
2 Nikolaev Institute of Inorganic Chemistry, Siberian Branch,
Russian Academy of Sciences, Novosibirsk, Russia
3 Diamond and Precious Metal Geology Institute, Siberian Branch,
Russian Academy of Sciences, Yakutsk, (Sakha) Yakutia, Russia
4 Joint Institute of Mechanical Engineering of the NAS of Belarus,
Minsk, Belarus Abstract. Popigai is the world largest crater produced by an impact event. Abundant graphite in the target rocks underwent martensitic transformation into
a mixture of high-pressure phases (an aggregate of nanometer cubic diamond
and hexagonal lonsdaleite crystals), and some amount of graphite survived as a
residual phase. They are of two types: (i) diamonds extracted from tagamites as
chips of grains crushed during processing; (ii) yakutites in placers inside and
around the crater, which formed at the impact epicenter and dispersed during the
event. The impact diamonds possess exceptional abrasive strength, 1.8 to 2.4
times greater than in synthetic diamonds. The outstanding wear resistance, a
large specific surface area and a thermal stability (200–250 °C greater than in
synthetic diamonds) are favorable for main technological uses. With these
properties, impact diamonds are valuable as material for composites and tools. Keywords: Impact crater Impact diamond Yakutite Abrasion strength © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 179–182, 2019.
https://doi.org/10.1007/978-3-030-22974-0_41 2
Popigai Impact Diamonds A huge crater, about 100 km in diameter, formed 35.7 Ma when a large bolid hit the
Earth at the Anabar shield in Siberia (Masaitis et al. 1998). The target Archean gneisses
of the Khapchan Group containing abundant crystalline graphite were broken, remolten
and partly dispersed outside the crater. Thereby the graphite underwent martensitic
transformation into a mixture of high-pressure cubic diamond and hexagonal lons-
daleite; some amount of graphite survived as a residual phase either in aggregates with
high-pressure phases or as separate particles. Upon conversion, the graphite reduced in
volume by 1.6 and the resulting diamond formed as aggregate of n * 10 – n * 100 nm
crystals (Walter et al. 1992). Impact diamonds possess exceptional abrasive strength,
greater than synthetic diamonds by 1.8 to 2.4 depending on relative percentages of
phases. The ultrahard impact diamonds can be successfully used for composites and
tools. 1
Introduction The Popigai impact crater located in Yakutia (Russia) stores unlimited amounts of
impact diamond with exceptional technological properties. Studies of the Popigai impact crater have a dramatic history. The research was
active from 1971, when V.L. Masaitis first proved the impact origin of the crater, to
1986, when the work stopped unexpectedly. Since neither geological surveys, nor
engineering testing were undertaken; the testing was impossible because few diamond
samples were available. In 2010 we got a collection of impact diamonds (about 3000
carat) from previous work at Popigai and resumed the tests. In this paper we charac-
terized the diamonds as valuable raw for advanced technologies. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 179–182, 2019. https://doi.org/10.1007/978-3-030-22974-0_41 180 V. Afanasiev et al. Impact Diamonds: Types, Properties and Uses We explored the Udarnoye and Skalnoye sites within the Popigai crater. The
Skalnoye deposit stores up to 100 carat diamond per ton of tagamite, while the total
estimated resources exceed 162 billion carat (Masaitis et al. 1998). The resources in the
whole crater are actually inexhaustible. The amount of yakutites in placers around the
crater may reach 1.5 billion carat (Masaitis et al. 1998). 3
Types of Impact Diamonds The Popigai diamonds comprise yakutites in placers and diamonds hosted by taga-
mites, the primary impact rocks. Yakutites formed at the impact epicenter and dispersed during the event. Currently
they are found in placers inside and around the crater. The farthest dispersed findings of
yakutites occur 550 km away from the crater or even more. The yakutite aggregates of
nanometer grains are 0.7–0.8 mm to 1.0 mm in size and have a shapeless morphology
or sometimes preserve hexagonal contours of primary graphite. They formed at the
highest pressure and consist of cubic diamond and hexagonal lonsdaleite. The presence
of graphite appears in Raman spectra but is undetectable by X-ray diffractometry,
possibly, because of minor contents. Tagamite-hosted diamonds are extracted by crushing from very hard host rocks and
thus have angular shapes and particle sizes from a few microns to 1 mm. During
subsequent flotation, large amounts of fine graphite are extracted along with diamond. These diamonds are likewise aggregates of nanometer diamond, lonsdaleite and gra-
phite grains. Diamond chips look laminated like the precursor graphite. Impact dia-
monds from tagamite are of two species. A-diamonds are colorless or yellowish, almost
fully consisting of the cubic phase; they are the most resistant to wear. Those of B
species comprise cubic diamond, lonsdaleite and graphite and are slightly less resistant. Most of impact diamonds, including those we analyzed, are from the Skalnoye deposit
in the southwestern flank of the Popigai crater, away from the epicenter. They formed
by shock waves while the target rocks were melting and remained enclosed in tagamite,
i.e. they originated at lower pressures than yakutites. Thus, placer yakutites and tagamite-hosted diamonds share the same impact origin
but formed at different pressures and evolved in different ways afterwards. The highest-
pressure yakutites were dispersed and quenched, and preserved their primary structure,
whereas diamonds in tagamites underwent prolonged annealing in tagamite melt. The
annealing explains the presence of nitrogen impurity (N3V) which lacks in yakutites. 181 Impact Diamonds: Types, Properties and Uses 5
Conclusions The Popigai impact diamonds possess exceptional abrasion strength, large specific
surface area and high thermal stability superior over the respective properties in syn-
thetic diamonds. With these technological properties and actually unlimited resources,
the impact diamonds are valuable industrial raw. The development of the Popigai
diamond deposit can be successful due to progress. Acknowledgements. The work was supported by grants 16-05-00873a and 17-17-01154a of the
Russian Foundation for Basic Researchers and was carried out as a part of the Project No. 0330-
2016-0006. 4
Possible Uses of Impact Diamonds Possible uses of impact diamonds are based on their exceptional wear resistance. Only
tagamite-hosted diamonds were studied. Micropowders were made from a collection of
impact diamonds (mixed A and B species) at the Institute of Superhard Materials
(Kyiv, Ukraine). The impact diamond micropowder of any grain size shows greater
abrasive strength than synthetic diamonds (Table 1). Table 1. Comparative abrasive strength of impact synthetic diamonds
Impact diamond micropowders
Synthetic diamond micropowders
Grain size, µm
Abrasive strength,
relative units
Grain size, µm
Abrasive strength,
relative units
+60
5,05
+60
3,67
60/28
6,53
60/28
3,69
40/20
5,89
40/20
3,54
28/14
5,70
28/14
3,33
14/7
4,85
14/7
2,91
10/5
3,80
10/5
2,16
7/3
2,98
7/3
1,71
5/2
2,20
5/2
1,21 Table 1. Comparative abrasive strength of impact synthetic diamonds The extremely hard impact diamonds can be superior substitutes for synthetic
diamonds in industry. Furthermore, a large specific surface area and relatively high
adsorption and thermal stability 200–250 °C greater than in synthetic diamonds, make
impact diamonds excellent raw for composites. Sintered composites of “impact dia-
mond - Fe-Ti bond” powdered in a planetary mill to 5 to 50 µm granules show better
polishing properties compared with similar powders based on synthetic diamond DSM
20/14 (Table 2). Table 2. Comparative polishing properties of impact diamonds – Fe-Ti bond and similar
powder of synthetic diamonds
Composite powders
Polished material Weight loss (mg/min) Stability (min)
Fe-Ti/impact diamond 5/50 Silica
35,4
>30
Fe-Ti/ACM
5/50 Silica
17,8
14 le 2. Comparative polishing properties of impact diamonds – Fe-Ti bond and similar
der of synthetic diamonds 182 V. Afanasiev et al. The impact diamond-based abrasive powders used for magnetic-abrasive polishing
of silica plates have 1.5–2 times greater strength and 2 times greater stability (lifetime)
than their counterparts made from synthetic diamonds (Table 2). The technological properties of yakutites remain poorly investigated. They obvi-
ously have a very high wear resistance, possibly, higher than in tagamitic diamonds,
while their grain sizes are suitable for using them in blade tools. Meanwhile, apart from abrasive properties, there is a large scope of potential uses
associated with structure, phase composition, etc. Currently, the only problem is the
shortage of diamond specimens for testing. Sufficient amounts of such specimens can
be obtained by building a pilot plant for extraction of diamonds. References Masaitis VL, Mashchak MS, Raikhlin AI, Selivanovskaya TV, Shafranovskiy GI (eds) (1998)
Diamond-bearing impactites from the popigai impact crater. VSEGEI, St. Petersburg
Walter AA, Eryomenko GK, Kvasnitsa VN, Polkanov Y (1992) Carbon minerals produced by
impact metamorphism. Naukova Dumka, Kiev Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Authentic Semi-precious and Precious
Gemstones of Turkey: Special Emphasis
on the Ones Preferred for Prayer Beads E. Çiftçi1(&), H. Selim2, and H. Sendir3 1 Department of Geological Engineering, Faculty of Mines, ITU, 33469 Maslak,
Istanbul, Turkey
eciftci@itu.edu.tr
2 Faculty of Engineering, Department of Jewellery Engineering, ITICU,
34840 Küçükyalı, Istanbul, Turkey
3 Faculty of Engineering and Architecture, Department of Geological
Engineering, EOGU, 26480 Odunpazarı, Eskişehir, Turkey 1 Department of Geological Engineering, Faculty of Mines, ITU, 33469 Maslak,
Istanbul, Turkey
eciftci@itu.edu.tr
2 Faculty of Engineering, Department of Jewellery Engineering, ITICU,
34840 Küçükyalı, Istanbul, Turkey
3 Faculty of Engineering and Architecture, Department of Geological
Engineering, EOGU, 26480 Odunpazarı, Eskişehir, Turkey Abstract. There are 6 semi-precious gemstones in Turkey that are the most
significant in terms of abundance and authenticity. These include smoky quartz,
blue
chalcedony,
chrysoprase
(aka
Şenkaya
emerald),
diaspore
(aka
sultanite/zultanite), sepiolite (aka meerschaum/Eskişehirstone), and jet (aka
Oltustone). The smoky quartz occurs in the south of Büyük Menderes Basin
within metamorphic rocks of the massif. Chalcedony occurrences are in
Çanakkale and Sarıcakaya (Eskişehir). Chrysoprase is acquired in Bursa, Ala-
şehir (Manisa) as yellowish green in color, Biga (Çanakkale), Sivrihisar (Eski-
şehir) and Şenkaya (Erzurum) as dark green in color. Diaspore occurs in Milas
(Muğla), Söke (Aydın), Tire (İzmir), Bolkardağı Gerdekkilise area and Saimbeyli
(Adana). Sepiolite occurrences are limited to Kıbrısçık-Köroğlu (Bolu), Eskişehir
and Konya. Jet (aka Oltustone) occurs in Oltu area (Erzurum). These are the
major varieties that has been used to produce both jewelry and ornamental
objects among which the prayer beads are one of the most indispensable object of
oriental cultures in particular. These are also considered to be the authentic gems
of Turkey and exported as raw and processed in into many forms. As for the
prayer beads, lightness, color, durability, hardness, and cost are the main criteria. Thus, the Oltustone has been the major source for decades. However, the Eski-
şehir stone and blue chalcedony are becoming popular as well. Keywords: Turkish gems Oltustone Eskişehirstone Diaspore
Smoky quartz Blue chalcedony Keywords: Turkish gems Oltustone Eskişehirstone Diaspore
Smoky quartz Blue chalcedony Keywords: Turkish gems Oltustone Eskişehirstone Diaspore
Smoky quartz Blue chalcedony © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 183–188, 2019.
https://doi.org/10.1007/978-3-030-22974-0_42 1
Introduction Today, the jewelry sector is living its golden age. With increasing technical capacity
and manual aptitude, gold, silver, platinum etc. metals can be embroidered with the
precious and semi-precious gems and delivered to consumers. This sector is a fashion-
driven dynamic sector and subjected to rapid changes and innovations that are con-
trolled and dictated by the fashion trends and demands of the consumers. Companies E. Çiftçi et al. 184 are forced to keep up with those and both develop and manufacture the products
accordingly. Use of the semi-precious and precious gems is likewise chosen according
to the demand by the fashion and modern trends. Consumers religiously follow those
two facts. Some of those are considered to be semi-precious gems of Turkey including
smoky quartz, chalcedony, chrysoprase, diaspore, sepiolite, and jet (Selim 2015). 2
Methods and Approaches Representative samples that acquired from the deposits were prepared for further
analytical studies for comprehensive characterization. Polished sections and powdered
samples were prepared for Transmitted and Reflected Light Microscopy integrated with
Optical Cathodoluminescence, Electron Probe Microanalysis (EPMA) and Secondary
Electron Microscopy-Energy Dispersive Spectroscopy (SEM-EDS), X-ray Diffraction
(XRD), X-ray Fluorescence (XRF) and Inductively-Coupled Plasma Mass Spec-
troscopy analyses. 3.3
Chrysoprase Chrysoprase or chrysoprasus is a gemstone variety of chalcedony that contains small
quantities of nickel. The color varies from faint light green to apple green (Fig. 2). It is
translucent and opaque. It worldwide occurrences are reported from USA (California,
Oregon, and Arizona) and Eastern India. In Turkey, it occurs in Alaşehir (Manisa) in
yellowish green, Dikmendağı area in Biga (Çanakkale) deep green, Sivrihisar (Eski-
şehir) in deep green, İkizce village (Bursa), Zümrüt and Turnalı villages of Şenkaya
county (Erzurum). In Erzurum, due to it locality and most probably to the color, it is
erroneously named as “Şenkaya Emerald”. It has naturally formed patterns and quasi
figures that make small objects even more fascinating. Fig. 2. Macroscopic views of natural chrysoprase and diaspore (Selim 2015) Fig. 2. Macroscopic views of natural chrysoprase and diaspore (Selim 2015) 3.2
Chalcedony Chalcedony occurs in a few locations including Bolu (Kıbrısçık), Ankara (Beypazarı),
Afyon (Bayat), İzmir (Aliağa, Bergama, Seferihisar-Yukarıdoğanbey), Tokat (Zile) in
lilac-purple colors, Yozgat (Çekerek) with amethyst as botryoidal masses, Çanakkale
(center) and Eskişehir (Sarıcakaya) in blue color. Druzy chalcedony occurs in Sivas
(Yıldızeli) (İlhan 2012). Chalcedony is a silica variety mineral. Its color may vary from
deep blue to faint blue, purple in places, translucent and opaque (Fig. 1). Among these,
blue chalcedony occurrence in Turkey is located in Mayıslar village (Sarıcakaya-
Eskişehir) owned and exploited by Kalsedon Co. (Hatipoğlu et al. 2010a). Blueish
color is either due to the water content or abnormally high barium content. This is also
considered to be the type locality. Its major use is as jewelry and ornamental objects. 3.1
Smoky Quartz Smoky quartz is one of the varieties of quartz with dark gray-black color (Fig. 1). It
occurs rarely in the Alps, Colorado and California (North America). Its occurrence in
Turkey is limited to the Menderes Massif, one of the metamorphic complexes of
Turkey. It occurs in metamorphic rocks composed of mafic minerals that expose in
southern portion of the massif in Great Menderes basin (İlhan 2012). Location is the
city of Aydın and covers a few rustic villages within Çine, Ovacık and Koçarlı county
limits. Host-rocks are variable degree metamorphic rocks including slate, phyllite,
schist, and gneiss. Smoky quartz occurs locally within the fractures of these rocks. Its
major use is as jewelry and ornamental objects. Type locality is Mersinbelen village in
the area. Fig. 1. Macroscopic views of natural smoky quartz and blue chalcedony (Selim 2015) Fig. 1. Macroscopic views of natural smoky quartz and blue chalcedony (Selim 2015) 185 Authentic Semi-precious and Precious Gemstones of Turkey 3.5
Sepiolite Sepiolite, also known as meerschaum, is a soft white clay mineral - a complex mag-
nesium silicate, a typical chemical formula (Mg4Si6O15(OH)26H2O). It can be present
in fibrous, fine-particulate, and solid forms. Sepiolite is opaque and off-white, grey or
cream color, breaking with a conchoidal or fine earthy fracture, and occasionally
fibrous in texture (Fig. 3). There many reports on its occurrence worldwide. It is
generally associated with carbonate/evaporate sequences, sedimentary in origin. However, it also occurs in Kıbrısçık (Bolu) as a result of hydrothermal alteration of
glassy tuffs of Middle Miocene Deveören volcanics in Köroğlu (Galatya) volcanic belt
(Irkeç and Ünlü 1993). Although there are occurrences worldwide, most of the sepiolite
of commerce is obtained chiefly from Sepetçi, Margı, Sarısu, Kayı, Gökçeoğlu, İmi-
şehir, and Türkmentokat villages of the city of Eskişehir in Turkey. Nemli (Kütahya) is
also important for its alike occurrences. It is mostly used to make pipes and small
objects, paring beads. Calcite, gypsum and dolomite often company sepiolite. Fig. 3. Macroscopic views of worked and raw Eskişehirstone and raw Oltustone (Selim 2015) g. 3. Macroscopic views of worked and raw Eskişehirstone and raw Oltustone (Selim 2015 3.4
Diaspore Diaspore is one of the minerals forming the bauxite ore. It is the least hydrous Al-
oxyhydroxide mineral in the bauxites. Diaspore becomes the main, even only mineral
of bauxites when the bauxite ore gets metamorphosed. Such ores are better called as
diasporite, since those are monomineralic. This is very common occurrence especially
in the Menderes Massif (western Anatolia). Diaspore crystals sometimes grow into
really gigantic sizes. Then becomes gem material. Diaspore naturally occur in yel-
lowish green and light green colors (Fig. 2). In addition to the metabauxites, diaspore E. Çiftçi et al. 186 occurrences are also reported in laterites and Al-rich clays, in recrystallized limestones,
with corundum, in some alkali pegmatites as late stage hydrothermal mineral, and in
magmatic rocks as a result of recrystallization of Al-rich xenoliths (Hatipoğlu et al. 2010b; Kumbasar and Aykol 1993). However, they hardly grow into gem size and
quality. In Turkey, type locality is Milas county (Muğla) (Lake Bafa vicinity and area
in between Milas-Yatağan counties). There are reports on its existence in Söke (Aydın),
Tire (İzmir), in central Anatolia (Gerdekkilise-Bolkardağ) and Saimbeyli county
(Adana), however, these are not at mineable scale. Gem quality diaspores are mostly
used in jewelry. Two names were adopted, also trademarks, for such diaspores,
“Sultanite or Zultanite” in honor of Ottoman Sultans and “Csarite”. It has very unique
phenomenal color changing property and rarity thus becoming more popular. It is
considered as “diamond of the future” by many. 4
Conclusions Among the many, 6 semi-precious stones are the most common and renowned: smoky
quartz, Eskişehirstone, Oltustone, sultanite, blue chalcedony, and Şenkaya emerald. Major use of these include jewelry (sultanite, Oltustone, smoky quartz, blue chal-
cedony, and Şenkaya emerald), ornamental objects (Eskişehirstone, Oltustone, smoky
quartz, blue chalcedony, and Şenkaya emrald), prayer beads (Oltustone, blue chal-
cedony, and Eskişehirstone). Sultanite (diamond of the future) is a rising star in the
precious stones due to its only occurrence in Milas area (Muğla) and unique properties. 3.6
Jet/Oltu Stone Jet (literal name) or Oltustone (locality name), is an opaque black coalified fossilized
drift wood of trees of the family Araucariaceae which is 180 million years old (Fig. 3). Authentic Semi-precious and Precious Gemstones of Turkey 187 It is formed from the high-pressure decomposition of wood. Jet is chemically related to
brown coal, or lignite, but it is more solid and tough, looks more like obsidian than a
coal (Toprak 2013). Jet has been used both as a talisman and a jewel for over four
thousand years. In Turkey, it occurs and mined in Oltu county (Erzurum) where more
than 300 locations around Tutlu (Lispek), Yeşilbağlar (Norpet), Güllüce, and Güzelsu
villages. Jet is also found in other locations around the world. However, only the
British and Spanish deposits in addition to the Turkish occurrences have been worked
commercially. Jet is called “Oltustone” in Turkey due to its type locality (Ciftci et al. 2004). It is mostly used in jewelry and to make praying beads. Framboidal pyrites and
clay pockets commonly occur in jet. Selim HH (2015) Türkiye’nin değerli ve yarı değerli mücevher taşları. İstanbul Ticaret Odası
(İTO) Yayınları No: 2014/4, 102s, İstanbul
Toprak S (2013) Petrographical properties of a semi-precious coaly stone, Oltu stone, from
eastern Turkey. Int J Coal Geol 120:95–101 References Ciftci E, Yalcin MG, Yalcınalp B, Kolaylı H (2004) Mineralogical and physical characterization
of the Oltustone, a Gemstone Occurring around Oltu (Erzurum-Eastern Turkey). In: 8th
ICAM, São Paulo-Brazil, pp 537–538 Hatipoğlu M, Babalık H, Chamberlain SC (2010a) Gemstone deposits in turkey. Rocks Miner 85
(2):124–133 Hatipoğlu M, Türk N, Chamberlain SC, Akgün AM (2010b) Gem-quality transparent diaspore
(Zultanite) in bauxite deposits of the Ilbir Mountains, Menderes Massif. SW Turk Miner
Deposita 45(2):201–205 İlhan NN (2012) Türkiye’nin Mücevher Taşları Haritası. I. Türkiye Mücevher Taşları
Sempozyumu, Bildiriler Kitabı, İstanbul, pp 33–38 İrkeç T, Ünlü T (1993) Volkanik kuşaklarda hidrotermal Sepiyolit oluşumuna bir örnek:
Kıbrısçık (Bolu) Sepiyoliti. MTA Dergisi, Ankara, vol 115, pp 99–118 Kumbasar I, Aykol A (1993) Mineraloji. İstanbul Teknik Üniversitesi Kütüphanesi, Sayı: 1519,
İstanbul 1
Introduction Copper and zinc are mainly extracted from sulfide ores using pyrometallurgical tech-
niques. Pyrometallurgical processing of arsenic containing ores is a problem due to the
emission of toxic gases (Filippou et al. 2007). Biohydrometallurgy is widely used to
process gold bearing concentrates, and may also be used to extract non-ferrous metals
arsenic-containing concentrates (Neale et al. 2017). The goal of the present work was to
study copper bioleaching from arsenic-containing minerals and chalcopyrite at different
temperatures and in NaCl presence. 2
Methods and Approaches Chalcopyrite (CuFeS2), enargite (Cu3AsS4), and tennantite (Cu12As4S13) as well as
mixed culture of acidophilic microorganisms oxidizing ferrous iron and sulfur com-
pounds were subjects of the study. The experiments were carried out in flasks with
100 ml of nutrient medium supplemented and 2 g of milled minerals (P100 75 µM) on a
rotary shaker at temperatures from 40 °C to 60 °C for 30 days. In one variant of the
experiment, nutrient medium was supplemented with100 mm NaCl. Biooxidation of Copper Sulfide Minerals Yu. Elkina1,2(&), E. Melnikova2, V. Melamud2, and A. Bulaev1,2 Yu. Elkina
, E. Melnikova , V. Melamud , and A. Bulaev
1 Faculty of Biology, Lomonosov Moscow State University, Moscow, Russia
yollkina@mail.ru
2 Research Center of Biotechnology RAS, Moscow, Russia 1 Faculty of Biology, Lomonosov Moscow State University, Moscow, Russia
yollkina@mail.ru
2 Research Center of Biotechnology RAS, Moscow, Russia Abstract. The effects of temperature and the presence of NaCl on bioleaching
of chalcopyrite, enargite, and tennantite were studied. Rate of copper extraction
from all minerals depended on temperature and was the highest at 45–50 °C. NaCl addition increased rate of copper extraction from chalcopyrite but led to
the decrease in copper extraction from enargite and tennantite. Keywords: Bioleaching Chalcopyrite Enargite Tennantite
Acidophilic microorganisms Selim HH (2015) Türkiye’nin değerli ve yarı değerli mücevher taşları. İstanbul Ticaret Odası
(İTO) Yayınları No: 2014/4, 102s, İstanbul
Toprak S (2013) Petrographical properties of a semi-precious coaly stone, Oltu stone, from
eastern Turkey. Int J Coal Geol 120:95–101
188
E. Çiftçi et al. E. Çiftçi et al. 188 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 4
Conclusions The results obtained suggest that the efficiency of copper sulfide minerals depended on
temperature, while NaCl addition did not allowed increasing the rate of copper
bioleaching from arsenic-containing minerals in contrast to chalcopyrite. This fact
should be taken into consideration when planning laboratory scale trials on bioleaching
of copper sulfide concentrates. Acknowledgements. The work was supported by the President Grant of the Russian Federation,
grant No. MK-6639.2018.8. 3
Results and Discussion The results of the experiments (rates of copper extraction) are shown in Table 1. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 189–191, 2019. https://doi.org/10.1007/978-3-030-22974-0_43 Yu. Elkina et al. 190 Table 1. Rate of copper extraction from the minerals for 30 days (%)
Variant of the
experiment
Chalcopyrite
(CuFeS2)
Enargite
(Cu3AsS4)
Tennantite
(Cu12As4S13)
40 °C
14.33 ± 0.08
8.18 ± 0.65
15.33 ± 0.24
45 °C
17.84 ± 1.69
12.89 ± 1.79
26.16 ± 1.33
50 °C
25.29 ± 4.15
14.04 ± 0.95
18.43 ± 0.84
50 °C, 100 NaCl
33.25 ± 0.12
5.91 ± 1.09
13.04 ± 0.03
55 °C
26.75 ± 1.57
14.39 ± 0.01
14.84 ± 0.01
60 °C
17.32 ± 1.01
5.86 ± 2.18
12.83 ± 0.17 Table 1. Rate of copper extraction from the minerals for 30 days (%) We showed that the rate of copper extraction from all minerals depended on
temperature and was low at 40 °C. Extraction rate at 60 °C was also low as this
temperature might inhibit microbial activity. Addition of NaCl increased rate of copper
extraction from chalcopyrite that was a well known phenomenon. At the same time,
NaCl addition led to the decrease in copper extraction rate. Neale J, Seppälä J, Laukka A, van Aswegen P, Barnett S, Gericke M (2017) The MONDO
minerals nickel sulfide bioleach project: from test work to early plant operation. Solid State
Phenom 262:28–32 1
Introduction The main resources of titanium in Russia are concentrated in identified (Pizhemskoe
and Yaregskoye deposits) and are designed (Vodnenskoe and other manifestations) of
the Devonian titanium paleoplacers Timan. Ores are represented by leucoxene, to a
lesser extent altered under exogenous conditions by ilmenite (pseudo-ethyl). Ore is
difficult to enrich due to the large number of quartz inclusions in titanium minerals. A large number of quartz inclusions in titanium minerals is an exceptional feature of
Timan paleoplacers. The high content of quartz in titanium minerals is explained by its
primary metamorphogenic genesis. Slates of the Precambrian folded basement of the
Timan are the root source of titanium minerals. Titanium paleoplacers are formed due
to the redeposition of the weathering crust on shale (Kochetkov 1967; Kalyuzhny 1972;
Makhlaev 2006; Ponaryadov 2017). Genetic Problem of Quartz in Titanium
Minerals in Paleoplacers of Middle Timan
I. Golubeva1, I. Burtsev1, A. Ponaryadov1, and A. Shmakova1,2(&) I. Golubeva1, I. Burtsev1, A. Ponaryadov1, and A. Shmakova1,2(&) 1 Institute of Geology Komi SC UB RAS, Syktyvkar, Russia
alex.sch92@yandex.ru
2 Karpinsky Russian Geological Research Institute (VSEGEI),
Saint-Petersburg, Russia 1 Institute of Geology Komi SC UB RAS, Syktyvkar, Russia
alex.sch92@yandex.ru
2 Karpinsky Russian Geological Research Institute (VSEGEI),
Saint-Petersburg, Russia Abstract. Titanium ore in the Devonian paleoplacers of Middle Timan is
predominantly represented by leucoxene, less frequently by modified ilmenite
(pseudorutile). Other titanium minerals are found in small amounts (or have a
sharply subordinate significance). All titanium minerals have numerous inclu-
sions of quartz, which create an intractable problem in the enrichment of titanium
ore. Metamorphogenic porphyroblastic explains the presence of quartz in titanium
minerals. Precambrian seric-chlorite clay weathering crusts are a supplier of
titanium minerals Timan. Leucoxene and ilmenite form in paraschist under con-
ditions of facies of green shale of regional metamorphism, poikiloblasts. In the
poikiloblasts, the poikilite and helicitic structures are well defined, due to
numerous poikilite inclusions of quartz. The poikiloblasts, poikilite and heliic
structures are well represented, due to the numerous poikilite incorporating quartz. Keywords: Titanium Paleoplacer Shale Poikiloblastez
Leucoxene Ilmenite Keywords: Titanium Paleoplacer Shale Poikiloblastez
Leucoxene Ilmenite References Filippou D, St-Germain P, Grammatikopoulos T (2007) Recovery of metal values from copper –
arsenic minerals and other related resources. Miner Process Extr Metall Rev 28(4):247–298 arsenic minerals and other related resources. Miner Process Extr Metall Rev 28(4):247 298
Neale J, Seppälä J, Laukka A, van Aswegen P, Barnett S, Gericke M (2017) The MONDO
minerals nickel sulfide bioleach project: from test work to early plant operation. Solid State
Phenom 262:28–32 Neale J, Seppälä J, Laukka A, van Aswegen P, Barnett S, Gericke M (2017) The MONDO
minerals nickel sulfide bioleach project: from test work to early plant operation. Solid State
Phenom 262:28–32 191 Biooxidation of Copper Sulfide Minerals Biooxidation of Copper Sulfide Minerals Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 192–194, 2019.
https://doi.org/10.1007/978-3-030-22974-0_44 Keywords: Titanium Paleoplacer Shale Poikiloblastez
Leucoxene Ilmenite Photos of minerals were taken on a JSM-6400 scanning microscope with a Link ISIS-
300 energy-dispersive spectrometer and a polarized microscope. 2
Methods and Approaches Photos of minerals were taken on a JSM-6400 scanning microscope with a Link ISIS-
300 energy-dispersive spectrometer and a polarized microscope. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 192–194, 2019. https://doi.org/10.1007/978-3-030-22974-0_44 193 Genetic Problem of Quartz in Titanium Minerals in Paleoplacers 3
Results and Discussion Titanium minerals - ilmenite and leucoxene (rutile, anatase and quartz aggregate)
crystallize the parachale under the conditions of regional metamorphism of the green
slate facies. This process is widely developed in the Riphean schists of Timan. The
content of titanium minerals varies from 1.5–3.0%, occasionally rising to 5%. Titanium
minerals crystallize in the form of porphyroblasts saturated with numerous quikilic
poikilite inclusions. Poikilite inclusions of quartz are fragments of aleurite sizes. The
quikite and inclusions of quartz and sericite captured during porphyroblasty determine
the helicocyte structure (Fig. 1a, b). Titanium minerals are easily separated from shale
and are separated during physical and chemical weathering. They accumulate due to
gravitational separation during transportation and redeposition of weathering products. In paleoplacer metamorphic structures of titanite in titanium minerals are well pre-
served (Fig. 1d, f). In titanium minerals, fragments of quartz veins recorded with the
growth of titanium minerals are diagnosed in paleoplacer. Sometimes fragments of
sericite-chlorite schists with ilmenite grains are found (Fig. 1e). The quartz inclusions
in leucoxene upon lithification of the ore-bearing sandstone can be regenerated with an
increase in the volume of inclusions. The amount of SiO2 in titanium minerals in
bedrock - shale is 6.6–11.47%, and in paleoplacer - 12.2–28.19% (Ignatiev 1997). The
high content of silicon dioxide in the titanium minerals of paleoplacer Timan is a
specific feature of ore in this area and is explained by the metamorphogenic porphy-
roblastic genesis. For comparison, the SiO2 content in titanium minerals of another
well-known Tuganov paleoplacer in Western Siberia is given: silica is 1.82% for Fig. 1. Titanium minerals in shale and paleoplacers: a - leucoxene porphyroblasts in Riphean
sericite-chlorite shale; b - helicitic structure in cross section of leucoxene porphyroblast in schist;
c - poikilite inclusions in the longitudinal section of porphyroblast ilmenite in schist; d - helicitic
structure in cross section leucoxene plate from paleoplacers; e - helicitic structure in leucoxene
from paleoplacers; f - a longitudinal section of ilmenite with poikilitite inclusions of quartz from
paleoplacers; g - a rounded fragment of sericite-chlorite shale with the inclusion of ilmenite from
paleoplacers Fig. 1. 3
Results and Discussion Titanium minerals in shale and paleoplacers: a - leucoxene porphyroblasts in Riphean
sericite-chlorite shale; b - helicitic structure in cross section of leucoxene porphyroblast in schist;
c - poikilite inclusions in the longitudinal section of porphyroblast ilmenite in schist; d - helicitic
structure in cross section leucoxene plate from paleoplacers; e - helicitic structure in leucoxene
from paleoplacers; f - a longitudinal section of ilmenite with poikilitite inclusions of quartz from
paleoplacers; g - a rounded fragment of sericite-chlorite shale with the inclusion of ilmenite from
paleoplacers I. Golubeva et al. 194 ilmenite and 8.5.4% for leucoxene 6.5. The primary metamorphogenic nature of tita-
nium minerals in Timan is indirectly indicated by the same increased content of MnO
in porphyroblasts of leucoxene and ilmenite in native shale (2.5–3.63%) and in pale-
oplacer (to 2.33%). 4
Conclusions The presence of a large amount of quartz in the titanium minerals of the paleoplacer
Timan is explained by the porphyroblastic growth of shales under conditions of
regional metamorphism of ilmenite and leucoxene. The increase in the percentage of
silica in titanium minerals is also due to the processes of lithogenesis. Acknowledgements. This research was supported by project AAAA-A17-117121270037-4
“Scientific basis for effective development and use of the mineral resource base, development and
implementation of innovative technologies, geological and economic zoning of the Timan-North
Ural region”. Ural region”. References Ignatiev VD, Burtsev IN (1997) Timan’s leucoxen: mineralogy and technology issues. Science,
St. Petersburg 215 p g
p
Kalyuzhny VA (1972) Geology of new placer formations. Science, Moscow 263 p Kalyuzhny VA (1972) Geology of new placer formations. Science, Moscow 263 p
Kochetkov OS (1967) Accessory minerals in the ancient strata of Timan and Kanin. Science,
Leningrad 200 p Kochetkov OS (1967) Accessory minerals in the ancient strata of Timan and Kanin. Science,
Leningrad 200 p Makhlaev LV, Golubeva II (2006) Ilmenite-containing metapelites as the most important source
of the formation of giant and supergiant titanium placers. Titanium-zirconium deposits of
Russia and prospects for their development. Moscow, IGEM RAS, pp 39–42 Ponaryadov AV (2017) Mineralogical and technological features of ilmenite-leucoxene ores of
Pizhemskoe deposit, Middle Timan. Vestnik of the Institute of Geology, Komi SC UB RAS,
no 1, pp 29–36. https://doi.org/10.19110/2221-1381-2017-1-29-36. (in Russian) Ponaryadov AV (2017) Mineralogical and technological features of ilmenite-leucoxene ores of
Pizhemskoe deposit, Middle Timan. Vestnik of the Institute of Geology, Komi SC UB RAS,
no 1, pp 29–36. https://doi.org/10.19110/2221-1381-2017-1-29-36. (in Russian) Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Gold and Platinum Group Minerals
(PGM) from the Placers of Northwest
Kuznetsk Alatau (NWKA)
(South Siberia, Russia) V. Gusev1,2(&), S. Zhmodik1,2, G. Nesterenko2, and D. Belyanin1,2 1 Department of Geology and Geophysics, Novosibirsk State University,
Novosibirsk, Russia
vityansky@igm.nsc.ru
2 Institute of Geology and Mineralogy SB RAS (IGM SB RAS),
Novosibirsk, Russia 1 Department of Geology and Geophysics, Novosibirsk State University,
Novosibirsk, Russia
vityansky@igm.nsc.ru
2 Institute of Geology and Mineralogy SB RAS (IGM SB RAS),
Novosibirsk, Russia Abstract. Native gold and PGM from NW Kuznetsk Alatau (NWKA) (South
Siberia, Russia) have been investigated. Applying the complex of advanced
analytical, geo-chemical, and statistical techniques permits determination of
motherlode types of noble mineralization (NM). For gold, it is mineralization of
three types: 1 – the gold-sulfide-quartz type associated with dykes of the basic
composition and fault-line zones (2), as well as the gold-skarn type (3). For PGM it is mineralization of the Ural-Alaskan (1) and the ophiolitic (2) types,
as well as multicomponent alloys associated with layered intrusions (3). The
presence of rims and inclusions is indicative of postmagmatic transformations of
the minerals. In the meantime, Au and PGM from NW Kuznetsk Alatau placers
retain genetic traits of motherlodes. Keywords: Alluvial placers Gold Platinum Group Elements (PGE)
Altai–Sayan folded area Kuznetsk Alatau Keywords: Alluvial placers Gold Platinum Group Elements (PGE)
Altai–Sayan folded area Kuznetsk Alatau © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 195–197, 2019.
https://doi.org/10.1007/978-3-030-22974-0_45 1
Introduction Au placers with PGM (0.1–0.2% and more by weight of Au) are widespread on the
territory of the Kelbes placer region NWKA. The placers were abandoned during last
centuries, but motherlode deposits have not been discovered here. Au-quartz, Au-
quartz-sulfide types, and Au-magnetite (skarn) have been discovered in NE and the
central Kuznetsk Alatau region, as well as in Salair range, and PGE have been revealed
in chromites and dykes of the basic composition. The contribution presents new data on
the morphology, micro-inclusions, change types, and the composition of native Au and
PGM from placers and their comparison with NM from motherlodes of adjacent
regions in the South Siberia. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 195–197, 2019. https://doi.org/10.1007/978-3-030-22974-0_45 V. Gusev et al. 196 2
Methods and Approaches Minerals were selected from heavy mineral concentrates under binocular microscope. An Axio Scope A1 (Carl Zeiss) microscope was used for determination of sizes and
morphology of NM particles, and then polished preparations were made from these
particles for microscopic investigations by methods of ore and electron microscopy. The composition and interrelations between minerals, micro-inclusions, and newly
formed phases were studied by SEM (MIRA 3 LMU, Tescan Ltd.), and EMP methods
(Camebax Micro) in the share use Center for Multielement Isotope Studies
(Novosibirsk). 3
Results and Discussion The examined gold has different morphologies (sizes, degrees of grain rounding, and
deformation), it is characterized by the wide range of variations in the fineness (from
720‰ to 1000‰), by different degrees of chemical change, contains inclusions of
quartz, magnetite, and clay minerals, and it is coated with material consisting of Fe
hydroxides. PGM are represented by ferroplatinum and rutheniridosmine associations, as well
as by small amount of sperrylite and Pd minerals. More than 65% of grains belong to
ferrous platinum and isoferroplatinum; about 30% are minerals of the Ru, Ir and Os
system and about 3% are Pd minerals. The degree of grain rounding of ferroplatinum is
higher than that of rutheniridosmines and sperrylite, but it is generally low which
indicates a short range of their transfer. As impurities, Cu, Rh and Pd are commonly
found, Au is found in rare cases. 4
Conclusions The identified features of native Au and PGM indicate the presence of several types of
motherlodes, and possibly, intermediate reservoirs, as well as various distances of
placers from sources of supply. The data obtained are compatible with the assumption
that input of gold from ledge ores of Au-quartz, Au-quartz-sulfide, and Au-magnetite
types widespread in the eastern part of the Kuztetsk Alatau and on the Salair range, as
well as from sediments of the Simonovskaya suite. The sources of PGM were rocks of
the Ural-Alaskan and ophiolite complexes, fragments of which (in particular the
Kaygadatsky massiv) have been established in NWKA. Gold and PGM from the Placers of NWKA 197 Acknowledgements. This work was supported financially by the RFBR (Grant 19-05-00464)
and government assignment (project No. VIII.72.2.3 (0330-2014-0016)). Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches The following research methods were used in the work: mineralogical description of
sections of boreholes, study of samples of rocks, slides and polished sections, micro-
probe analysis of transparent polished thin sections from ore intervals, statistical
treatment of chemical analysis data with program Statistica v.6.1 and then analysis and
interpretation of results. These methods allowed to identify similarities and differences
in the ore intervals and their mineralization, and to confirm the assumption of a various
genesis of PGE mineralization in the ore zone “C” (Subbotin et al. 2012). © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 198–200, 2019.
https://doi.org/10.1007/978-3-030-22974-0_46 1
Introduction Since the late 1980s in Kola Peninsula the platinum content, associated with ultramafic-
mafic magmatic complexes, has been studied. The Fedorovo-Pana early proterozoic
layered intrusion of the peridotite-pyroxenite-gabbronorite formation has been recog-
nized as the most promising object for discovering the industrial reserves of complex
low sulfide PGE ores. Noble Metal Mineralization of the PGM Zone
“C” of the East-Pana Layered Intrusion
(Kola Peninsula) O. Kazanov1, G. Logovskaya2(&), and S. Korneev2 O. Kazanov1, G. Logovskaya2(&), and S. Korneev2 1 Moscow Branch, FSUE “All-Russian Scientific-Research Institute of Mineral
Resources named after N.M. Fedorovsky”, Moscow, Russia
2 Institute of Earth Science, Saint-Petersburg State University,
Saint-Petersburg, Russia
galkanuu@gmail.com Abstract. The first study of the noble metal mineralization of the ore zone “C”
of the East-Pana massif allowed to divide it into two types: early magmatic low-
sulfide and late post-magmatic proper PGE associations. Keywords: Fedorovo-Pana massif East-Pana massif
Stratiform mineralization Platinum metal mineralization
Early magmatic and late post-magmatic platinum mineral associations Keywords: Fedorovo-Pana massif East-Pana massif
Stratiform mineralization Platinum metal mineralization
Early magmatic and late post-magmatic platinum mineral associations Keywords: Fedorovo-Pana massif East-Pana massif
Stratiform mineralization Platinum metal mineralization Early magmatic and late post-magmatic platinum mineral associations Early magmatic and late post-magmatic platinum mineral associations 3
Results and Discussion Based on the results of the study of ore mineralization within the ore zone “C”, two
associations of noble metal minerals were identified. The first, early, magmatic asso-
ciation is represented by platinum and palladium sulfides (Fig. 1), native gold and Noble Metal Mineralization of the PGM Zone “C” 199 silver, moncheite, kotulskite and ferroplatinum, forming inclusions in the magmatic
major sulphide and rock-forming silicate minerals. There is a significant positive
correlation dependence of Pt, Pd, Au with Cu, Ni and S. It is indicated in the Sungiyok
area. Later, postmagmatic association is represented by sperrylite (Fig. 1), mertieite-I,
mertieite-II, temagamite, telargpalite, stibiopalladinite and kotulskite, associated with
secondary silicate and later sulphide (millerite) minerals. They develop along cracks
and veins in the rock-forming and early sulfide minerals. For the second type, corre-
lations with Cu are not characteristic, with S there is a significant negative correlation. This association is observed in the Chuarvy area. Both types of mineralization are
characterized by abnormally low values of Pd/Pt = 0.1–1.7 (Voytekhovich et al. 2008). Fig. 1. Subhedral zonal aggregate of braggite ((Pt, Pd, Ni)S - early magmatic association of
PGE mineralization (left); Spot edge around amphibole and micron streak of sperrylite (PtAs2) -
postmagmatic, hydrothermal metasomatic association of PGE mineralization (on the right). Fig. 1. Subhedral zonal aggregate of braggite ((Pt, Pd, Ni)S - early magmatic association of
PGE mineralization (left); Spot edge around amphibole and micron streak of sperrylite (PtAs2) -
postmagmatic, hydrothermal metasomatic association of PGE mineralization (on the right). 4
Conclusions Thus, based on the identified features of the noble metal mineralization of the ore
horizon “C”, it can be divided into two types: low-sulfide platinum and properly
platinum. The first type is characterized by a close relationship with sulfide minerals:
noble metal minerals form intergrowths with the main sulphides, are included in rock-
forming silicate minerals. This type is typical for the ore horizons of the Sungiyok area. Minerals of noble metals are localized in cracks and streaks and are associated with
minerals formed during the post-magmatic stage of intrusive evolution. This type is
observed in Sunghiyok area and Chuarvy area. Acknowledgements. Authors are grateful to V.V. Shilovskich for the research of minerals
which carried out on the “Hitachi S-3400 N” scanning electron microscope at “Geomodel”
Resource Centre of Saint-Petersburg State University. O. Kazanov et al. 200 Subbotin VV, Korchagin AU, Savchenko EE (2012) Platinometal mineralization of the
Fedorovo-Pana ore cluster: types of mineralization, mineral composition, genesis peculiar-
ities. Vestnik Kola Sci Cent Russ Acad Sci 1:55–65 Subbotin VV, Korchagin AU, Savchenko EE (2012) Platinometal mineralization of the
Fedorovo-Pana ore cluster: types of mineralization, mineral composition, genesis peculiar-
ities. Vestnik Kola Sci Cent Russ Acad Sci 1:55–65
Voytekhovich VS, Kazanov OV, Kalinin AA (2008) Report on the results of prospecting and
assessment work on platinum metal mineralization in the eastern part of the PanaTundras-
massif in 2006–2008. LLC Kola Mining and Geological Company, Apatity Voytekhovich VS, Kazanov OV, Kalinin AA (2008) Report on the results of prospecting and
assessment work on platinum metal mineralization in the eastern part of the PanaTundras-
massif in 2006–2008. LLC Kola Mining and Geological Company, Apatity Institute of Geology, Karelian Research Centre, Petrozavodsk, Russia
shchipts@krc.karelia.ru Abstract. Shungite rocks are widespread in Zaonezhye, Republic of Karelia,
where they constitute dozens of carbonaceous rock deposits of the Paleopro-
terozoic Onega structure with predicted carbon resources of more 4 billion tons. The lower age boundary is of 2.1 Ga. Shungite rocks belong to carbonaceous
rock class. These rocks metamorphosed in greenshcist facies of muscovite-
chlorite-biotite subfacies are unique natural, noncrystalline, non-graphitized,
fullerene-like
carbon. They have various
structural-mineralogical
levels:
(a) supramolecular, (b) molecular, (c) electron-energetic, (d) structural-physical
and (e) geologic-genetic (parametric). Shungite rocks contain shungite carbon
(shungite matter) and a variety minerals, microminerals and nanominerals. The
applications of shungite rocks are determined with regard for their natural types. Authors had shown their intergrated application in ore-thermal processes. Keywords: Shungite rocks Carbon Metamorphism Paleoproterozoic
Application Shungite rocks, named so after a Karelian town, Shunga, have attracted attention for
decades and have no counterparts in the Earth’s geological evolution with respect to the
mode of occurrence and tremendous reserves. They are unique from both scientific and
practical points of view. Shungite rocks are part of Paleoproterozoic carbonaceous
formation in Karelia. They are common in the Trans-Onega area, Russian Karelia,
where 251010 tonnes of autochthonous organic matter have been formed over an area
of about 9000 km2. These complexes constitute dozens of carbonaceous rock deposits
in the Paleoproterozoic Onega Structure with forecast carbon reserves of over 4 billion
tones and are mainly confined to the rocks of the Ludicovian system with the lower age
boundary of 2.1 Ga. The bulk of free carbon (Corg.) is in the Trans-Onega suite of the
Ludicovian superhorizon. Phenomenal carbon accumulation in this superhorizon is
responsible for the mineralogenic specialization of the rocks of the Trans-Onega suite. Their Corg. concentration varies from less than 1% to 70 wt% in rocks and up to
98 wt% in anthraxolite aggregates. Carbon concentration in the rocks of the Kondopoga
suite of the Kalevian superhorizon is not more than a few percent. Thus, the black shale
formation of the Onega Structure is formed of the rocks of the Trans-Onega, Suisari and
Kondopoga suites. The world’s largest Zazhogino Ore Field, covering an area of
З240 km2, with two active quarries, Zazhogino and Maksovo, has been identified and
relevant evidence was presented (Table 1). Metamorphism and metasomatism have contributed markedly to the genetically
distinctive shungite rocks. They belong to a metamorphogenetic class of shungite
formation. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 201–204, 2019.
https://doi.org/10.1007/978-3-030-22974-0_47 References Subbotin VV, Korchagin AU, Savchenko EE (2012) Platinometal mineralization of the
Fedorovo-Pana ore cluster: types of mineralization, mineral composition, genesis peculiar-
ities. Vestnik Kola Sci Cent Russ Acad Sci 1:55–65 Voytekhovich VS, Kazanov OV, Kalinin AA (2008) Report on the results of prospecting and
assessment work on platinum metal mineralization in the eastern part of the PanaTundras-
massif in 2006–2008. LLC Kola Mining and Geological Company, Apatity Voytekhovich VS, Kazanov OV, Kalinin AA (2008) Report on the results of prospecting and
assessment work on platinum metal mineralization in the eastern part of the PanaTundras-
massif in 2006–2008. LLC Kola Mining and Geological Company, Apatity Voytekhovich VS, Kazanov OV, Kalinin AA (2008) Report on the results of prospecting and
assessment work on platinum metal mineralization in the eastern part of the PanaTundras-
massif in 2006–2008. LLC Kola Mining and Geological Company, Apatity Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Shungites and Their Industrial Potential V. Kovalevski and V. Shchiptsov(&) Institute of Geology, Karelian Research Centre, Petrozavodsk, Russia
shchipts@krc.karelia.ru Institute of Geology, Karelian Research Centre, Petrozavodsk, Russia
shchipts@krc.karelia.ru Their industrial properties are controlled by metamorphism and a special V. Kovalevski and V. Shchiptsov 202 Table 1. Comparative description of the parameters of major deposits in Zazhogino Ore Field
Ore body shape
Size
Maximum
thickness
Average free carbon
content, %
Length, m
Width, m
Shunga deposit
Sheet-like
1400
300
5.2
41
Maksovo deposit
Sheet-like-cone
shaped
700
500
120
40
Zazhogino deposit
Sheet-like-cone
shaped
400
300
60
27
Zalebyazhskoe deposit
Sheet-like
2000
700
38
35 Comparative description of the parameters of major deposits in Zazhogino Ore Field contribution of K-Na alkaline metasomatism which has affected the formation of the
natural types and varieties of shungite and, correspondingly, industrial types and
varieties and applications of this unique raw material. On the P-T scheme, showing
principal correlations between metamorphic facies and subfacies after S.A. Bushmin
and V.A. Glebovitsky, shungite rocks were derived under greenschist-facies,
muscovite-chlorite-biotite-subfacies conditions of metamorphism at a temperature of
325–450 °C and a pressure of 2–5 kbar (Bushmin and Glebovitsky 2016). The mineral form of shungite (shungite matter) is non-graphitizable fullerene-like
carbon, which differs from graphite at a supramolecular, atomic and zonal (electron)
structure level. The main supramolar character of shungite is an ability to form
spherical structures (empty globules). At an atomic level, in addition to hexagonal rings
alone typical of graphite, pentagonal and heptagonal rings, characteristic of fullerene-
like structures, are also observed. At a zonal structure level, the energy of collective
excitations of valent (external) and framework (internal) p- and r-electrons decreases
relative to graphite, which is typical of fullerenes as well (Kovalevsky et al. 2016). Shungite from some deposits displays diamagnetic properties characteristic of full-
erenes. The structure of shungite rocks is similar to that of vitreous-crystalline mate-
rials, where highly dispersed crystals are distributed in non-crystalline matrix. Shungite rocks belong to a class of carbonaceous rocks varying in carbon content
and mineral diversity and those are natural carbon-mineral composite materials. They
contain shungite (shungite matter) and a variety of micro- and nanominerals. Silicate
minerals are highly disperse and are evenly distributed in carbon matrix. Major rock-
forming minerals are quartz, mica, albite and pyrite. High secondary and accessory
mineral concentrations and a certain spectrum of layered and cluster impurities are
observed. Natural types of shungite rocks display several textural and structural vari-
eties
and
the
non-uniform
phase
composition
of
carbon
and
geochemical
characteristics. Institute of Geology, Karelian Research Centre, Petrozavodsk, Russia
shchipts@krc.karelia.ru Shungite applications are determined by natural types and varieties of shungite
rocks (Kovalevsky et al. 2016). The structure and properties of shungite rocks are Shungites and Their Industrial Potential 203 responsible for their application in oxidation-reduction processes: in blast furnace
production of foundry (high-silicon) cast iron: in ferroalloy production; in yellow
phosphorus production; in carbide and silicon nitride production; as a reinforcing
component of groove masses; as a filler of non-stick paints. The sorptive catalytical and reduction properties of shungite rocks are used: for
treatment of high-quality drinking water in flow systems and wells; for removal of
many contaminants from urban domestic and industrial sewage; for swimming-pool
water treatment; for water treatment at heat power plants; for electrically conductive
paint production; for electrically conductive concrete and brick production; for elec-
trically conductive and plastering and masonry solutions; for electrically conductive
asphalt production. These materials were used for developing and designing heaters:
rooms screening electromagnetic radiation; a method for removal of ice from roads
(warm pavements and roads). Finely ground shungite can be mixed with any binders of organic and inorganic
origin and can thus be used as: black pigment for oil and water paints; a filler for
polymers materials (polyethylene, polypropylene, fluoroplastic, etc.); substitute for
white soot and technical carbon in rubber production. One essential feature of shungite rocks is that they can be modified with regards for
their desirable application. Enrichment of shungite rocks and their division into micro-
and nanosized components make a possible to activate shungite carbon and expand the
potential applications of shungite rocks in science-intensive technologies, e.g. nanotechnologies. There is only one geologo-industrial classification of shungite rocks (after Yu. Kalinin) based on industrial types distinguished with respect to their mineralogical
composition and corresponding applications. However, the latest results of the large-
scale practical application of shungite rocks shows that such a division into industrial
types is clearly insufficient. Therefore, division into subtypes or varieties is required. At the present stage of the study of Karelia’s shungite rocks the goal of research
and appraisal is not only to estimate the reserves of potential deposits and to assess the
petrographic and structural-chemical characteristics of shungite rocks but to develop
criteria and recommendations for the industrial application of rocks from potential
deposits in innovative and science-intensive fields (Kalinin and Kovalevski 2013). For this purpose it is proposed: 1. Institute of Geology, Karelian Research Centre, Petrozavodsk, Russia
shchipts@krc.karelia.ru To re-assess earlier geological, physico-chemical and technological data on known
deposits to make up a list of known shungite rocks suitable for particular
applications. 1. To re-assess earlier geological, physico-chemical and technological data on known
deposits to make up a list of known shungite rocks suitable for particular
applications. 2. To reveal promising shungite rock applications (e.g. metallurgy, tyre production,
water disposal, the production of composite materials, etc.) and certification
requirements to be met by raw material for each application. 3. To specify the critical properties of shungite rocks for each application (e.g. chemical composition, mineralogical composition, the structural parameters of
carbon and rocks) and express methods for their analysis. 4. To develop certification requirements for shungite rocks with regard for their
applications. 204 V. Kovalevski and V. Shchiptsov 5. To analyze shungite rock samples and cores from each of the bodies and rock
exposures and classify all shungite rocks into types and subtypes referenced to a
major application, based on the criteria developed. 5. To analyze shungite rock samples and cores from each of the bodies and rock
exposures and classify all shungite rocks into types and subtypes referenced to a
major application, based on the criteria developed. 6. To correlate the geological parameters of the deposits and outcrops with the geo-
chemical and structural-petrological characteristics of shungite rocks to forecast the
prospecting of shungite rocks with preset properties for required applications. 6. To correlate the geological parameters of the deposits and outcrops with the geo-
chemical and structural-petrological characteristics of shungite rocks to forecast the
prospecting of shungite rocks with preset properties for required applications. A classification of the geologo-industrial types of shungite rocks will be worked out
and the most promising shungite rock prospects for each application will be specified,
based on the results of prospecting and prospecting-and-appraisal for shungite rocks. The accomplishment of the work planned will result in the efficient investment of the
money spent in the cost of future deposits and the updating of innovative approaches to
the use of Russia’s unique carbonaceous raw material. Shungite has unveiled many of his mysteries, has become known all over the
world, has attracted the interest of experts by its great potential and yet has remained
largely unknown and open to new discoveries. Acknowledgements. The work is performed in the framework of the PFNI GAN research of IG
KarRC RAS. References Bushmin SA, Glebovitsky VA (2016) Scheme of mineral facies of metamorphic rocks and its
application to the Fennoscandian Shield with representative sites of orogenic gold
mineralizaion. Trans KRC RAS 3–27 Bushmin SA, Glebovitsky VA (2016) Scheme of mineral facies of metamorphic rocks and its
application to the Fennoscandian Shield with representative sites of orogenic gold
mineralizaion. Trans KRC RAS 3–27 Kalinin Yu, Kovalevski V (2013) Shungite rocks: scientific search horizons. Nauka v Rossii
6:66–72 Kalinin Yu, Kovalevski V (2013) Shungite rocks: scientific search horizons. Nauka v Rossii
6:66–72 Kovalevsky V, Shchiptsov V, Sadovnichy R (2016) Unique natural carbon deposits of shungite
rocks of Zazhogino ore field, Republic of Karelia, Russia. In: SGEM Conference Proceedings
of International Multidisciplinary Scientific GeoConference Surveying Geology and Mining
Ecology Management, pp 673–680 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 205–207, 2019.
https://doi.org/10.1007/978-3-030-22974-0_48 Gold-Silver Natural Alloy of Chromitites
from the Kamenushinsky Massif
(The Middle Urals) A. Minibaev(&)
Mining University, Saint-Petersburg, Russia
a.m.minibaev@yandex.ru A. Minibaev(&) Mining University, Saint-Petersburg, Russia
a.m.minibaev@yandex.ru Abstract. A detailed study of the chromitites clinopyroxenite Kamenushinsky
massif in the middle Urals has allowed to allocate from them the gold-silver
alloy. The study of the chemical composition, morphology of gold-silver alloys,
peculiarities of their placement allowed not only to determine their genetic
relationship with platinum-bearing chromites, but also to make an assumption
about the formation of chromitites as a result of a single geological process. Keywords: Urals Platinum Belt Kamenushinskiy massif
Chromite-platinum mineralization Gold Keywords: Urals Platinum Belt Kamenushinskiy massif
Chromite-platinum mineralization Gold 2
Methods and Approaches A study of morphology and chemical composition of Au-Ag natural alloys and its
bearing chromitites was carried out by the scanning electron microscope Carl
Zeiss EVO (OPTEC, Moscow), equipped with attachment: EDS (energy dispersive X-
ray spectrometer) and BSD (Backscattered Electron Detector). 1
Introduction The history of the development of platinum placers of the Urals dates back nearly a two
centuries and a distinctive feature is the permanent presence of gold and its natural
alloys along with the extraction of the platinum groups minerals (hereafter - PGM)
(Vysotsky 1913). It is known that the main source of the PGM is a process of a zonal
massifs erosion of the Ural-Alyaskan type. Nevertheless, the question of a primary
source of gold in placers is remained as a controversial issue. Thus, for example, in
research works of N. Vysotsky (Vysotsky 1913) was given a support of the idea that
the appearance of gold in platinum placers was due to the weathering of an acid rocks,
but its absence in some zonal massifs of the Urals Platinum Belt (hereafter - UPB)
keeps the matter in abeyance. In the 70s of the 20th century during setting deposit into
exploitation, located within the Kachkanar zonal massif UPB, for the first time gold
was determined and described from the bed rock (Fominykh et al. 1970). In spite of this
fact, further the question of the nature of the appearance of gold in the ores of the UPB
massifs was not under active discussion. Inside the Nysyamsky platinum placer field – one of the main leaders of the
platinum mining of the Urals – also the presence of gold was reported (Vysotsky 1913). The source of the placer formations was Kamenushinsky dunite-clinopyroxenite massif
which had a potential for of bed chromite-platinum mineralization (Ivanov 1997;
Tolstykh et al. 2011, Minibaev et al. 2015). During the study platinum-bearing
chromitites were divided into two petrographic types with characteristic features: vein-
imbedded and massive (Minibaev 2018). In addition to study of PGM features from the A. Minibaev 206 chromitites of both petrographic types, the scientific observation of the determined Au-
Ag alloys is under a great interest 4
Conclusions The obtained results are correlated with the conclusions about the syngenetic nature of
the platinum-bearing vein-imbedded and massive chromitites, where the latter are
characterized by a later origin relating to the final stage of evolution of the ore-forming
system (Minibaev 2018). Also these results can be used not only to confirm the
hypothesis about the common substance source of two petrographic types of chromi-
tites, but also allow to make the conclusion that the formation of chromitites is gen-
erally resulted from a single geological process. Acknowledgements. Work
was
done
with
the
support
of
government
program
№
5.12856.2018/8.9. 3
Results and Discussion Au-Ag natural alloys was determined as impurities of irregular crystallographic habitus
in chromespinelides of vein-imbedded (Fig. 1a) and massive (Fig. 1b) chromitites. The size of the aggregates is around 4–6 microns. The chemical composition of Au-
Ag alloys (Table 1) corresponds to the compositions of similar objects founded in the
ore chromospinelides of the Konder massif (Pushkarev et al. 2015). The similar composition of Au-Ag alloys, its presence directly in chromespinelides,
also the absence of traces of visible deformations of the mentioned indicates that its
formation does not correspond to the processes of serpentinization or overlaid
hydrothermal processes (as it was known before), it corresponds to the fact that
inclusions were gained into chromespinelides during crystallization. Inclusions such as
Cr and Fe in Au-Ag alloys are the evidence of inherited condition of mineralization and
chemistry features of the ore-forming system. Table 1. Chemical composition of Au-Ag alloys from the chromitites of the
Kamenushinsky massif (mass. %)
№point
Au
Ag
Cr
Fe
Sum
1
81,45
10,84
3,84
2,02
98,15
2
81,21
10,47
3,55
1,89
97,12
Notes: 1 - from vein-imbedded chromitites; 2 - from massive chromitites. 1. Determination of grains Au-Ag natural alloys in chromitites: a - vein-imbedded; b -
v
Table 1. Chemical composition of Au-Ag alloys from the chromitites of the
Kamenushinsky massif (mass. %)
№point
Au
Ag
Cr
Fe
Sum
1
81,45
10,84
3,84
2,02
98,15
2
81,21
10,47
3,55
1,89
97,12
Notes: 1 - from vein-imbedded chromitites; 2 - from massive chromitites. Table 1. Chemical composition of Au-Ag alloys from the chromitites of the
Kamenushinsky massif (mass. %) Fig. 1. Determination of grains Au-Ag natural alloys in chromitites: a - vein-imbedded; b -
massiv Fig. 1. Determination of grains Au-Ag natural alloys in chromitites: a - vein-imbedded; b -
massiv Determination of grains Au-Ag natural alloys in chromitites: a - vein-imbedded; b - 207 References Fominykh VG, Uskov ED, Volchenko YuA (1970) Gold in the ores of the Kachkanar massif in
the Middle Urals. In: Questions of the Geology of the Gold Deposit, Tomsk, pp 31–36 Ivanov OK (1997) Concentric-zonal pyroxenite-dunite massifs of the Urals. Ed. Ural University,
Ekaterinburg, 488 p Minibaev AM (2018) About the origin of chromite-platinum mineralized zones of the
clinopyroxenite-dunite Kamenushinsky massif of the middle Urals. In: Materials of the
Eighth Russian Youth School with International Participation “New in the Knowledge of the
Processes of Ore Formation”. IGEM RAS, Moscow, pp 255–258 Minibaev AM, Stepanov SYu (2015) Perspectives of the identification of chromite-platinum
mineralization in the rocks of the Kamenushinsky zonal clinopyroxenite-dunite massif (Middle
Ural). In: Proceedings of the Fifth Russian Youth School with International Participation “New
in the Knowledge of the Processes of Ore Formation”. IGEM RAS, Moscow, pp 145–148 Pushkarev EV, Kamenetsky VS, Morozova AV, Hiller VV, Glavatskikh SP, Rodemann T (2015)
Ontogeny of ore chromespinelide and composition of inclusions as an indicators of
pneumatolyte-hydrothermal formation of platinum-bearing chromite of the Konder massif
(Aldan shield). Geol Ore Deposits 57(5):394–423 Tolstykh ND, Telegin Y, Kozlov AP (2011) The native platinum of the Svetloborsky and
Kamenushinsky massifs of the Ural platinum-bearing belt. Geol Geophys 52(6):775–793
Vysotsky NK (1913) Platinum deposits of Isovsky and Nizhne-Tagilsky regions of the Urals. In:
Proceedings of the Geological Committee. New edit, no. 62, 692 p Tolstykh ND, Telegin Y, Kozlov AP (2011) The native platinum of the Svetloborsky and
Kamenushinsky massifs of the Ural platinum-bearing belt. Geol Geophys 52(6):775–793
V
k NK (1913) Pl i
d
i
f I
k
d Ni h
T
il k
i
f h U l
I Vysotsky NK (1913) Platinum deposits of Isovsky and Nizhne-Tagilsky regions of the Urals. In:
Proceedings of the Geological Committee. New edit, no. 62, 692 p Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. References Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Microbial Processes in Ore-Bearing Laterite
at the Tomtor Nb-REE Deposit: Evidence
from Carbon Isotope Composition
in Carbonates V. Ponomarchuk(&), E. Lazareva, S. Zhmodik, N. Karmanov,
and A. Piryaev
Institute of Geology and Mineralogy SB RAS, Novosibirsk, Russia
ponomar@igm.nsc.ru V. Ponomarchuk(&), E. Lazareva, S. Zhmodik, N. Karmanov,
and A. Piryaev Institute of Geology and Mineralogy SB RAS, Novosibirsk, Russia
ponomar@igm.nsc.ru Institute of Geology and Mineralogy SB RAS, Novosibirsk, Russia
ponomar@igm.nsc.ru Abstract. The unique Nb-REE deposit is located within the Tomtor complex of
ultramafic alkaline and carbonatite rocks in the northern Sakha Republic
(Yakutia) (Kravchenko and Pokrovsky 1995; Dobretsov and Pokhilenko 2010;
Lazareva et al. 2015). Ores reside in three layers (Severny, Yuzhny, and Buranny
sites) which fill depressions in subsided profiles of weathered carbonatites. Judging by stable isotope analysis, carbonates from laterite weathering profiles at
the Tomtor Nb-REE-deposit formed by different mechanisms, including micro-
bially mediated organic-clastic sulfate reduction and anaerobic oxidation of
methane. Keywords: Tomtor Nb-REE-deposit Laterite Carbonates C-isotopes
Microbial Methane Keywords: Tomtor Nb-REE-deposit Laterite Carbonates C-isotopes
Microbial Methane © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 208–211, 2019.
https://doi.org/10.1007/978-3-030-22974-0_49 2
Methods and Approaches The stable isotope compositions were analyzed in three samples (TM-592, TM-590,
and BH 101) from laterite ore zones with *1%, 0.6%, and 1.6% REE, respectively. Extraction of monofractions for analysis is difficult because the laterite samples are fine
grained while the carbonates are zoned. The problem can be solved by using selective
acid extraction based on progressively slower reactions of carbonates with H3PO4 in
the stoichiometric series: calcite ! dolomite ! ankerite ! siderite ! rhodochrosite. According to experimental evidence (Al-Aasm et al. 1990), the reaction duration
sufficient for d13C determination is 1 h for calcite, 24 h for dolomite, and 5–7 days for
siderite and rhodochrosite. The C and O isotope compositions of carbonates are
determined by digestion in anhydrous H3PO4 followed by measurements on a FIN-
NIGAN MAT-253 mass spectrometer with a GasBench II analyzer, in a stream of pure
helium. The GasBench unit was also used for preconditioning of samples digested at
70 °C. The results are quoted in (‰) relative to the Vienna Peedee belemnite PDB
standard for carbon and relative to SMOW for oxygen. The analytical errors were
±0.1‰ for carbon and 0.15‰ for oxygen. The composition and morphology of min-
erals were studied at the Analytical Center of IGM (Novosibirsk) on a Tescan MIRA 3
LMU scanning electron microscope with Oxford Instruments Nanoanalysis Aztec
Energy/INCA Energy 450+ XMax 80 and INCA Wave 500 analyzers, applicable to
scan nanometer particles. 1
Introduction Rare earth elements have been broadly used in advanced technologies. They often occur
in carbonatite deposits among which laterite profiles of weathered carbonatites are most
attractive commercial targets. A unique Nb-REE deposit is located within the Tomtor
complex of ultramafic alkaline and carbonatite rocks in the northern Sakha Republic
(Yakutia) (Kravchenko and Pokrovsky 1995; Dobretsov and Pokhilenko 2010; Lazar-
eva et al. 2015). Ores reside in three layers (Severny, Yuzhny, and Buranny sites) which
fill depressions in subsided profiles of weathered carbonatites. Lately, evidence has been
obtained that localization of elements in ore-bearing beds may have biotic controls
(Lazareva et al. 2015). The possible role of microorganisms in ore formation has not
been discussed yet in the literature on laterite profiles from carbonatite deposits,
including the giants of Mountain Weld (Australia) or Araxa and Catalão (Brazil). In this
respect, the Tomtor deposit is remarkable by a contribution of biogenic and bacterial
processes to its formation, besides magmatism and high-temperature hydrothermalism
that are common to all carbonatite deposits. Biogenic agents are usually identified from
the carbon isotope composition of rocks controlled by their interaction with fluids. The
discussed stable isotope compositions of carbonates from ore-bearing laterites at the
Tomtor deposit provide evidence of possible microbial mediation. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 208–211, 2019. https://doi.org/10.1007/978-3-030-22974-0_49 209 Microbial Processes in Ore-Bearing Laterite 3
Results and Discussion Sample TM-590 is composed mainly of goetite, siderite (with a Mn impurity), calcite,
and rhodochrosite. Goethite occurs either as dripstone, with concentric zonation, or as
pseudomorphs after a disappeared mineral. The carbonates exist as zoned anhedral
grains, with calcite being the latest phase. Fine apatite crystals, pyroxene, monazite,
pyrochlore, and sphalerite are present in lesser amounts. The rock-forming phases in
TM-592 are goethite, siderite, and calcite and the accessories are apatite and monazite. Apatite occurs as fine prismatic euhedral grains in calcite. Sample BH 101 consists of
finely intergrown Fe-chlorite (chamosite) and apatite. The apatite-chamosite aggregate
encloses clearly seen large rhodochrosite grains with siderite in their core. The two
phases have a distinct boundary but have inherited orientations of the crystallographic
axes. The accessory phases are TiO2, submicrometer monazite platelets, sphalerite, and
galena. Synthesis of C and O isotope data from magmatic and postmagmatic (hydrother-
mal, metasomatic) carbonates in different carbonatite deposits world wide shows a
large range of values from −10 to +3‰ (PDB) for d13C and from +6 to +30‰
(SMOW) for d18O (Deines 1989). The d13C values obtained for the Tomtor samples (Table 1) are lower. The d13C
patterns in carbonates are controlled by the isotope composition of bicarbonate. Low
d13C in bicarbonate is due to oxidation of organic matter where d13C can reach −32‰
(Oleary 1988). In the course of microbially mediated sulfate reduction of geopolymers
and biopolymers in aerobic conditions, the d13C values of bicarbonate that forms by V. Ponomarchuk et al. 210 reaction (1) and those of carbonate produced later by reaction (2) are inherited from the
precursor component. reaction (1) and those of carbonate produced later by reaction (2) are inherited from the
precursor component. 2CH2O þ SO2
4
! 2HCO3 þ H2S
ð1Þ
2HCO3 þ Me2 þ ¼ MeCO3 þ CO2 þ H2O
ð2Þ ð1Þ ð2Þ There are two facts that implicitly support the above considerations: laterite con-
tains organic remnants (Lazareva et al. 2015) that are electron donors in reaction (1);
the samples contain sulfides, including framboidal pyrite, which formed at the account
of H2S released in reaction (1). However, this model can account only for the isotope
composition of the calcite component in TM-590, while other d13C values (−24.8‰ in
TM-590) are below −32‰, and may have formed with participation of methane. 3
Results and Discussion Biogenic methane has −55‰ to −80‰ d13C, and the d13C values in bicarbonate
resulting from microbially mediated methane oxidation (react/3) and in carbonate that
formed later by reaction (2) will be low. CH4 þ SO2
4
! HCO3 þ HS þ H2O
ð3Þ ð3Þ Table 1. d13С and d18О (‰) in different fractions of carbonates
Sample
1 h
24 h
7 days
TM-590
d13С
−29.6
−39.2
−24.8
d18О
+12.0
+16.1
+14.6
TM-592
d13С
−31.2
−44.1
−37.1
d18О
+7.6
+13.6
+15.1
BH 101
d13С
−59.0
−54.6
–
d18О
+9.4
10.5
–
Note that a low value of d13C was previously reported (Pokrovsky 1990;
Kravchenko and Pokrovsky 1995) at the Tomtor deposit, but the mineralogical
and geochemical characterization of the sample is not given. Table 1. d13С and d18О (‰) in different fractions of carbonates Note that a low value of d13C was previously reported (Pokrovsky 1990;
Kravchenko and Pokrovsky 1995) at the Tomtor deposit, but the mineralogical
and geochemical characterization of the sample is not given. Note that a low value of d13C was previously reported (Pokrovsky 1990;
Kravchenko and Pokrovsky 1995) at the Tomtor deposit, but the mineralogical
and geochemical characterization of the sample is not given. Microbial Processes in Ore-Bearing Laterite 211 4
Conclusions The reported isotope data show that organic clastic sulfate reduction and anaerobic
methane oxidation with participation of microbial communities were among key
mechanisms responsible for the formation of carbonates in laterite profiles of the
Tomtor Nb-REE deposit. Acknowledgements. The study was supported by grant 18-17-00120 from the Russian Science
Foundation. Microbial Processes in Ore-Bearing Laterite References Al-Aasm IS, Taylor BE, South B (1990) Stable isotope analysis of multiple carbonate samples
using selective acid extraction. Chem Geol Isot Geosci Sect 80(2):119–125 de Toledo MCM, de Oliveira SMB, Fontan F, Ferrari VC, de Parseval P (2004) Mineralogia,
morfologia e cristaloquímica da monazita de Catalão I (GO, Brasil). Braz J Geol 34(1):135–
146 Deines P (1989) Stable isotope variations in carbonatites. In: Bell K (ed) Carbonatites, Genesis
and Evolution. Unwin Hyman, London, pp 301–359 Dobretsov NL, Pokhilenko NP (2010) Mineral resources and development in the Russian Arctic. Russ Geol Geophys 51(1):98–111 Kravchenko SM, Pokrovsky BG (1995) The Tomtor alkaline ultrabasic massif and related REE-
Nb deposits, Northern Siberia. Econ Geol 90(3):676–689 p
( )
Lazareva EV, Zhmodik SM, Dobretsov NL et al (2015) Main minerals of abnormally high-grade
ores of the Tomtor deposit (Arctic Siberia). Rus Geol Geophys 56(6):844–873 Lazareva EV, Zhmodik SM, Dobretsov NL et al (2015) Main minerals of abnormally high
ores of the Tomtor deposit (Arctic Siberia). Rus Geol Geophys 56(6):844–873 Olary MO (1988) Carbon isotopes in photosynthesis. Bioscience 38(5):328–336 Pokrovsky BG, Belyakov AYu, Kravchenko SM et al (1990) Origin of carbonatites and ore-
bearing rocks of the Tomtor massif, NW Yakutia, according to isotopic data. Geokhimiya
(9):1320–1329. (in Russia) Ribeiro CC, Brod JA, Junqueira-Brod TC, Gaspar JC, Petrinovic IA (2005) Mineralogical and
field aspects of magma fragmentation deposits in a carbonate–phosphate magma chamber:
evidence from the Catalão I complex, Brazil. J South Am Earth Sci 18(3–4):355–369 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 212–213, 2019.
https://doi.org/10.1007/978-3-030-22974-0_50 1
Introduction The increased interest to peridot – jewelry varieties of olivine, was observed over the
last years. This has resulted in numerous publications on the geology, mineralogy, and
genesis of this gem. Peridot has been found in almost 30 countries and on all conti-
nents, including Antarctica (gem-quality mineral is present in the basalts from the Ross
Island); furthermore it was diagnosed in several stony-iron meteorites. Peridot is pre-
sent in different geological formations, which almost all (with the exception of
exogenous placers) belong to derivatives of endogenous processes and often geneti-
cally related to ultrabasic and basic rocks. S. Sokolov(&) S. Sokolov(&) FSBE “All-Russian Institute of Mineral Raw Materials” (VIMS),
Moscow, Russia
vims-sokol@mail.ru Abstract. The generalized information about the peridot from deposits and ore
occurrence of well-known formation type is presented in this paper. Peridots
from specific deposits vary in the accompanying mineral associations. These
gems, depending on the genesis, contain differing in phase composition inclu-
sions of mineral-forming medium and have different crystallization temperatures. Keywords: Peridot Formations Deposits Origin Temperature offormation Peridot: Types of Deposits and Formation Conditions Placer deposits. Eluvial-deluvial plasers are the most productive among the
exogenous deposits of peridot. They are connected with different genetic types of
primary deposits of this gem (Zabargad, San-Carlos, Kugda). 3
Origin and Formation Conditions of Peridot Peridots from the deposits of different formation clearly differ in the associations of
accompanying minerals. A detailed study of the jewelry peridots from the Kovdor and Kugda deposits
allowed us to determine the specific features of their micromineralogy, (Yarmishko and
Sokolov 2005): crystalline inclusions, represented mainly by minerals accompanying
to peridot; decomposition products (magnetite dendrites and magnetite-diopside plates;
crystallized melt inclusions. Fixation multiphase crystallized melt inclusions in peridots confirms the magmatic
nature of the deposits some formation types. For example, olivine and calcite from
kimberlites of the Udachnaya-Vostochnaya pipe contain the melt inclusions the total
temperature range of homogenization of which is 1100–880 °C (Tomilenko et al. 2009). Peridots from basalts of the Hawaiian Islands contain the inclusion of glass (Gubelin and
Koivula 1992). The melt inclusions in jewelry peridot from the Kovdor deposits were
homogenized at temperatures of 970–930 °C. Crystallization of peridot at the Zabargad
deposit occured in temperature range from 900 to 750 °C (Maaskant 1986). 2
Formation Types of the Deposits of Peridot Diamond-bearing kimberlites, concentrating peridot in different quantities, are known
in Yakutia (Udachnaya-Vostochnaya and Mir pipes), the Republic of South Africa
(Kimberly and De Beers pipes), and in Tanzania (Mvadun pipe). Ultrabasic-alkaline rocks and carbonatites – UAC complexes. Peridot installed at
complexes of the Kola peninsula (Kovdor) and Polar Siberia (Kugda). Basalts and basaltoids (normal and (sub) alkaline). Peridot phenocrysts in the
nodules of peridotites and olivenites are situated in lava flows and volcanic craters at
deposits and ore occurrence in Russia, USA, Hawaiian Island, the Czech Republic,
Madagascar Island, Ethiopia, China, Pakistan, Australia, Antarctica. Alpinotype hyperbasites. The most famous deposit of peridot – Zabargad Island is
classed to this formation type. They are known also in Russia at the East Sayan and at
the South Urals. 213 Peridot: Types of Deposits and Formation Conditions References Gűbelin EJ, Koivula JI (1992) Photoatlas of inclusions in gemstones. ABC Edition, Zurich Gűbelin EJ, Koivula JI (1992) Photoatlas of inclusions in gemstones. ABC Edition, Zurich
Maaskant P (1986) Electron probe microanalyses of unopened fluid inclusions, semiquantitative
approach. Neues Jahrb Miner 7:297–304 Tomilenko AA, Kovyazin SV, Dunlyansky YuV, Pokhilenko LN (2009) Primary melt and fluid
inclusions in minerals from kimberlites of the Udachnaya-Vostochnaya pipe, Yakutia. ECROFI-XX. Abstracts, University of Granada, pp 255–256 Tomilenko AA, Kovyazin SV, Dunlyansky YuV, Pokhilenko LN (2009) Primary melt and fluid
inclusions in minerals from kimberlites of the Udachnaya-Vostochnaya pipe, Yakutia. ECROFI-XX. Abstracts, University of Granada, pp 255–256 Yarmishko SA, Sokolov SV (2005) Micromineralogy of peridots from rocks of alkaline-
ultrabasic massifs. In: VII International Conference «New Ideas in Earth Sciences». Abstracts,
vol 2, Moscow, p 76. (in Russian) Yarmishko SA, Sokolov SV (2005) Micromineralogy of peridots from rocks of alkaline-
ultrabasic massifs. In: VII International Conference «New Ideas in Earth Sciences». Abstracts,
vol 2, Moscow, p 76. (in Russian) Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction Mineralogical analysis is the most significant method in reconstructing sources pale-
oplacers and glacial deposits (boulder loams), determining location of source for glacial
provinces. The paper presents two objects - glacial deposits of polar and vychegda
horizons, Middle Devonian and Middle Triassic titanium paleoplacers. Mineralogical Analysis of Glacial Deposits
and Titanium Paleoplacers of the
East European Part of Russia N. Vorobyov and A. Shmakova(&) Institute of Geology of Komi SC UB RAS, Syktyvkar, Russia
alex.sch92@yandex.ru Abstract. Mineralogical analysis is one of the main methods to determine
sources for paleoplacers and location of source glacial provinces. Our studies
defined location of source glacier provinces and sources for titanium paleo-
placers of the East European Part of Russia. Keywords: Minerals Reconstruction Glacial deposits Titanium paleoplacers © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 214–215, 2019.
https://doi.org/10.1007/978-3-030-22974-0_51 2
Results and Discussion Quaternary deposits have been studied in two areas of Pechora lowland: in the
northeast in the basin of the Padymeytyvys river and northwest in the basin of the Kui
river. Titanium paleoplacers were studied at the Kydzarasyu river (Preural Foredeep)
and Middle Timan (Pizhma paleoplacer). Relations between mineral composition of Quaternary deposits and underlying
bedrocks are very important. During active exaration activity of the glacier, the
underlying rocks controlled composition of boulder loams. The formation of heteroaged
horizons of boulder loams is associated with the Northeastern (Paykhoy-Ural-Novaya
Zemlya) and Northwestern (Fennoscandinavia) terrigenous-mineralogical source pro-
vinces. The rocks of the eastern province are characterized by higher levels of epidote
and ilmenite fraction, and the northwestern ones - amphiboles and garnets. In the basin of the Padymeytyvys river two horizons of boulder loams were drilled. In the lower (Vychegodsky) horizon, epidote (19.9%) and siderite (21.2%) dominate in
the heavy fraction. In the composition of the heavy fraction of the upper (polar)
horizon, pyrite (22.1%) and epidote (20.7%) have maximum concentrations, siderite
content (11.7%) and ilmenite (14.7%) are high. Pyrite and siderite indicates the relation
between the glacier and Triassic and Permian underlying rocks, and ilmenite may be
associated with Uralian rocks. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 214–215, 2019. https://doi.org/10.1007/978-3-030-22974-0_51 215 Mineralogical Analysis of Glacial Deposits and Titanium Paleoplacers In the valley of the Kui river one horizon of boulder loam – polar – was drilled. The
heavy fraction is represented by amphibole (13.6%) - garnet (18%) - epidote (20.8%)
associations with the increased content of pyrite (7.7%). High concentrations of
amphiboles (13.6%) and garnets (18%) may indicate the relation between the glacier
and the rocks of Fennoscandinavia. The number of amphiboles in boulder loams decreases from west to east and
amounts to the first percent. High total contents of pyrite and siderite in the north-east
of Pechora lowland are typical of local Mesozoic source province. Heavy fraction of the paleoplacer at the Kydrasyu river is represented by ilmenite
(45.19%), epidote (23.21%), magnetite (15.76%), amphibole (6.69%), chrome spine-
lide (5.65%), garnets (2.06%). Also zircon, leucoxen, rutile, hematite, kyanite, martite,
pumpellyite, staurolite are present (0.04–0.4%). The high content of ilmenite indicates
the relation between the paleoplacer and igneous rocks. Epidote, garnet, kyanite,
staurolite, and pumpellyite found in the heavy fraction are of metamorphic origin. 2
Results and Discussion The
morphology of zircons is characteristic of minerals formed in igneous rocks of medium
composition. The peculiarity of these minerals is their good preservation. This may be
due to the nearby source. The mineral composition indicates that the source of minerals
was not far from its burial. Titanium minerals and satellite minerals were most likely of
magmatic and metamorphic origin. The mineral composition of titanium placers often contains stable minerals and
there are no unstable ones, although both are present in the bedrocks. The main reason
is that the paleoplacer formed due to erosion of the weathering crust. An example of a
paleoplacer formed that way can be Pizhma paleoplacer (Middle Timan) formed due to
erosion and redeposition of weathering crusts on Riphean shales. It’s mineral com-
position in contrast to the paleoplacer at the Kydzarasyu river, is very poor. The main
part is represented by leucoxenized ilmenite and leucoxene. Tourmaline, garnet and
zircon are found in single units. 3
Conclusions We determined location of the source glacier provinces and sources for the titanium
paleoplacers of the East European part of Russia by the mineralogical analysis. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction Nowadays, a global trend of oil fields with increased residual oil saturation is observed
(Drozdov and Drozdov 2012). There is also problem with associated petroleum gas
(APG) utilization, which is not used rationally and is directed to gas flares. Currently,
one of the most promising areas in oil production is the simultaneous water and gas
injection (SWAG) technology. The definite advantage of the SWAG is the ability to use
APG as a liquid-gas mixture compound. This method makes it possible to mount simple
maintaining, reliable and efficient equipment, while providing a significant reduction in
power consumption and increasing the SWAG efficiency (Drozdov et al. 2012). A Bench Scale Investigation of Pump-Ejector
System at Simultaneous Water
and Gas Injection S. Karabaev(&), N. Olmaskhanov, N. Mirsamiev, and J. Mugisho
Department of Mineral Developing and Oil and Gas Engineering,
Engineering Academy, RUDN University, Moscow, Russia
simpleforfiza@mail.ru S. Karabaev(&), N. Olmaskhanov, N. Mirsamiev, and J. Mugisho Department of Mineral Developing and Oil and Gas Engineering,
Engineering Academy, RUDN University, Moscow, Russia
simpleforfiza@mail.ru Abstract. In this paper, a bench study of the pump-ejector system for simul-
taneous water and gas injection (SWAG) was conducted. For these purposes, a
pump-ejector system stand was used. A differential pressure gauge was used to
determine the gas flow at the ejector intake. According to the results of differ-
ential manometer calibrations, a new formula was obtained which reduces its
inaccuracy to 1% at pressures below 0.6 MPa. In addition, according to the
pressure-energy diagrams, it was determined that the gas injection with excess
pressures in the ejector suction chamber significantly increases the efficiency of
the pumping-ejector system overall. Keywords: Liquid-gas ejector Pump-ejector system Liquid-gas mixture
SWAG © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 219–220, 2019.
https://doi.org/10.1007/978-3-030-22974-0_52 2
Methods and Approaches The investigation was conducted based on bench tests. The electric centrifugal pump
and the liquid-gas ejector were used, which represent a single bench-model of the
pump-ejector system. This stand is designed to investigate the characteristics of model
ejectors, multistage centrifugal pumps and pump-ejector systems with liquid-gas
mixtures using fresh water as a liquid, and air as a gas. The differential pressure gauge
was used to determine the gas flow, with allowance for the excess pressure created in S. Karabaev et al. 220 the suction chamber of the ejector. For operations at low pressures, the differential
pressure gauge was pre-calibrated before. 4
Conclusions Further study of the pressure-energy characteristics behavior and the causes affecting
this change will facilitate the increase of efficiency of technological processes which
use the pumping-ejector systems. 3
Results and Discussion Calibration of the differential manometer made it possible to clarify the existing for-
mula for determining the gas flow rate at the ejector suction chamber at pressures below
0.6 MPa. The inaccuracy of the calculated values obtained by this method was less
than 1%. According to the results of the conducted research, the pressure-energy
characteristics of the ejectors were constructed during gas suction by the liquid. There
occurs a significant expansion of liquid-gas ejector working range by the injection of
gas with excess pressure in the ejectors suction chamber as well as the areas of
maximum values of injection coefficients and efficiency rates are shifted to higher
region of operating pressure. References Drozdov AN, Drozdov NA (2012) Laboratory researches of the heavy oil displacement from
Russkoye field’s core models at the SWAG injection and development of technological
schemes of pump-ejecting systems for the water-gas mixtures delivering. In: SPE 157819,
presented at the SPE heavy oil conference canada held in Calgary, Alberta, Canada, pp 12–14
Drozdov NA, Drozdov AN, Malyavko EA (2012) Investigation of SWAG injection and
prospects of its implementation with the usage of pump-ejecting systems at existing oil-field
infrastructure. In: SPE 160687, presented at the SPE Russian oil and gas exploration and
production technical conference and exhibition held in Moscow, Russia, pp 16–18 Drozdov AN, Drozdov NA (2012) Laboratory researches of the heavy oil displacement from
Russkoye field’s core models at the SWAG injection and development of technological
schemes of pump-ejecting systems for the water-gas mixtures delivering. In: SPE 157819,
presented at the SPE heavy oil conference canada held in Calgary, Alberta, Canada, pp 12–14 schemes of pump-ejecting systems for the water-gas mixtures delivering. In: SPE 157819,
presented at the SPE heavy oil conference canada held in Calgary, Alberta, Canada, pp 12–14
Drozdov NA, Drozdov AN, Malyavko EA (2012) Investigation of SWAG injection and
prospects of its implementation with the usage of pump-ejecting systems at existing oil-field
infrastructure. In: SPE 160687, presented at the SPE Russian oil and gas exploration and
production technical conference and exhibition held in Moscow, Russia, pp 16–18 Drozdov NA, Drozdov AN, Malyavko EA (2012) Investigation of SWAG injection and
prospects of its implementation with the usage of pump-ejecting systems at existing oil-field
infrastructure. In: SPE 160687, presented at the SPE Russian oil and gas exploration and
production technical conference and exhibition held in Moscow, Russia, pp 16–18 Drozdov NA, Drozdov AN, Malyavko EA (2012) Investigation of SWAG injection and
prospects of its implementation with the usage of pump-ejecting systems at existing oil-field
infrastructure. In: SPE 160687, presented at the SPE Russian oil and gas exploration and
production technical conference and exhibition held in Moscow, Russia, pp 16–18 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. 1
Introduction The complex development of deposits and the most complete use of the resource
potential of the developed deposits, as well as the multifunctional use of the waste
space of mineral deposits, is one of the main tasks of mining science (Vorob’ev et al. 2017). In a number of oil fields, the screen is a salt layer of various capacities located
above the productive oil reservoir, which can be used to produce mineralized water
with subsequent injection of mineralized water into the productive oil reservoir, and the
production-capacity (underground reservoir) formed in the salt reservoir can serve as a
storage of hydrocarbons extracted from the productive reservoir (natural gas, associated
oil gas, oil). Integrated Use of Oil and Salt Layers at Oil
Field Development V. Malyukov and K. Vorobyev(&) Department of Mineral Developing and Oil & Gas Engineering,
Engineering Academy, RUDN University, Moscow, Russia
k.vorobyev98@mail.ru Department of Mineral Developing and Oil & Gas Engineering,
Engineering Academy, RUDN University, Moscow, Russia
k.vorobyev98@mail.ru Abstract. Combined development of oil and salt layers of the oil field allows to
obtain a mineralized solution to intensify the extraction of oil from the reservoir
and create an underground reservoir in the salt reservoir for underground storage
of hydrocarbons, including the creation of an underground gas storage (UGS) in
rock salt. Keywords: Oil reservoir Salt formation Field Combined use References Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 221–222, 2019.
https://doi.org/10.1007/978-3-030-22974-0_53 2
Methods and Approaches To obtain mineralized water, it is advisable to drill wells on the salt formation and
conduct underground dissolution of rock salt to obtain a solution of rock salt of a
certain concentration for injection through injection wells into the productive formation
and extraction of oil from the productive formation through producing wells. Pumping more fresh water into the clay-containing collector than the reservoir
water reduces the permeability of the collector and makes it a low-permeable collector. Controlling the mineralization of injected water and the properties of clays in pro-
ductive formations can significantly increase the oil recovery rate. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 221–222, 2019. https://doi.org/10.1007/978-3-030-22974-0_53 222 V. Malyukov and K. Vorobyev 4
Conclusions Complex use of oil and salt layers of the oil field allows to obtain a mineralized
solution to intensify the extraction of oil from the reservoir and to create an under-
ground reservoir in the salt reservoir for underground storage of associated petroleum
gas (APG), i.e. to create an underground gas storage (UGS) in rock salt. At Talakan oil and gas condensate field (Republic of Sakha) the implementation of
combined development of oil and salt layers of oil and gas condensate field with the
creation of an underground gas storage in rock salt and the use of mineralized solution
for the intensification of oil extraction was started. 3
Results and Discussion The use of mineralized water can reduce the hydration of formation clays, but it is
desirable to select the composition of water that is most compatible with the formation
components of the productive formation. The use of mineralized water obtained by dissolving the salt layer of the oil field
can largely solve the problem of compatibility of pumped mineralized water with
reservoir water and mineral composition of the reservoir (Lyashenko et al. 2018). When developing a salt formation with a capacity of several tens of meters with the
supply of solvent through the drilling well, according to the technological regulations,
an underground production is created-a container in rock salt (vertical or horizontal) for
storage of petroleum gas along the way. References Lyashenko V, Vorob’ev A, Nebohin V, Vorob’ev K (2018) Improving the efficiency of blasting
operations in mines with the help of emulsion explosives. Min Miner Deposits 1:95–102
Vorob’ev A, Chekushina T, Vorob’ev K (2017) Russian national technological initiative in the
sphere of mineral resource usage. Rudarsko Geolosko Naftni Zbornik. 2:1–8 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Oil and Gas Reservoirs in the Lower
Triassic Deposits in the Arctic Regions
of the Timan-Pechora Province N. Timonina(&)
Institute of Geology Komi SC UB RAS, Syktyvkar, Russia
nntimonina@geo.komisc.ru N. Timonina(&) Institute of Geology Komi SC UB RAS, Syktyvkar, Russia
nntimonina@geo.komisc.ru Abstract. The Lower Triassic sediments of the Timan-Pechora oil and gas
bearing province were studied by the complex of lithological, petrophysical and
geochemical methods. It is established that productive deposits are represented
by various-grained sandstones that formed in arid climates in the vast alluvial-
lacustrine plain. The research allowed to study and characterize the structure
features of rock and mineral aggregates, forming a pore space. The high
heterogeneity of the composition and structure of the cement minerals of the
reservoir caused by local facies-paleogeographic sedimentation conditions, were
the cause of the significant variability of the pore space. Rocks reservoirs are
complex, with high content of clay component, an effective development of
which requires special methods of stimulation. Keywords: Oil and gas bearing province Lithological types Reservoir
Clay minerals Pore space Porosity Permeability Keywords: Oil and gas bearing province Lithological types Reservoir
Clay minerals Pore space Porosity Permeability © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 223–226, 2019.
https://doi.org/10.1007/978-3-030-22974-0_54 2
Methods and Approaches Oil and gas reservoirs are geological bodies consisting of reservoir beds, lenses, and
reservoirs of weakly and impermeable rocks of intra-reservoir tires, forming a single
hydrodynamic system, bounded below and above by inter-reservoir tires. Accumula-
tion of hydrocarbons in the reservoir and their safety are determined by the quality of
each of the elements. The main reason for the differentiation of natural reservoirs by
properties is their formation. The structural features of sedimentary layers determine
the patterns of distribution of collectors and seals in them, their interrelationship, and
ultimately predetermine the morphology and properties of natural reservoirs. This paper proposes a conceptual model of the formation of a natural reservoir
confined to the deposits of the Charkabozhskaya suite. The construction of working
models was carried out using the results of complex processing of all available
information, including well-logging, and study of core material. The results of this work are based on combination of analytical tools, including
polarizing microscope, scanning electron microscope, X-ray diffraction and electron
microprobe, were used in order to identify and estimate authigenic mineral type sand to
determine paragenetic sequences. 1
Introduction In recent years, researches were intensified in the field of the formation of natural
reservoirs. The basis for sedimentological reconstructions lies in the idea that the
morphology and filtration characteristics of natural reservoirs are largely predetermined
by ancient sedimentation situations, which are closely associated with the tectonic
history of the territories. The Timan-Pechora province occupies the region of the
northeastern Russian platform. Hydrocarbon accumulation occurred in Triassic rocks
largely in the northern part of the basin, where pools were found in the rocks in the
Varandey, Toravey, Labagan fields of the Sorokin swell, Kumzhinskoe, Korovinskoe –
in Denisov Depression. Triassic deposits are distributed almost throughout the Timan-
Pechora oil and gas bearing province, with the exception of the axial zones of large
positive structures. The Lower Triassic includes strata of the Charkabozhsky suite,
which thickness varies from the first meters in the southwest (in the Seduyahinsky
swell) to 380 m in the central part of the Kolvinsky megaswell, the Khoreyver
depression, the average thickness of the suite is 150–250 m (Morakhovskaya 2000). The sediments are represented by rhythmic alternation of red-brown clays, greenish-
gray siltstones and gray sandstones with conglomerates and gravelites. Sandstones are
characterized by a variety of granulometric composition from fine to coarse-grained, as
well as a wide range of textures: massive, diagonal and horizontal bedding. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 223–226, 2019. https://doi.org/10.1007/978-3-030-22974-0_54 N. Timonina 224 N. Timonina Oil and Gas Reservoirs in the Lower Triassic Deposits Dominant cement minerals in Triassic sandstones include calcite, kaolinite,
smectite, illite, and chlorite. Carbonate cement include pure calcite and siderite. The
clear transparency and delicate crystalline habit of most of the clay, as revealed by
microscope and SEM analysis, leads us to conclusion that the most clays in these
sandstones are authigenic. Kaolinite occurs mostly as pore-lining and pore-filling
cement, some of them appear as visible alteration of detrital feldspars. The distribution
of kaolinite is uneven both in the section and in area. In the transition from coarse to
fine-grained sediments, a decrease in the content of kaolinite is noted. Chlorite is
common as pore-filling and lining cement. Smectite occurs both as grain-coating and
filling pores. The studied deposits are characterized by a wide distribution of smectite
minerals with increasing content upward the section. In the fine-grained sandstones of
the basal stratum, its content does not exceed 50–60%, whereas in the upper part of the
section its amount increases to 80–90%. The distribution and amount of clay minerals
in sandstone cement is determined by both conditions of sedimentation and post-
sedimentation transformations. The most widespread collectors are of III–V classes
according to the A.A. Khanin classification (Khanin 1976). Class VI reservoirs are
characteristic of floodplain formations and represented by aleurolites and fine-grained
sandstones, in which large pore channels are practically absent. Class V collectors are
represented by fine-grained sandstones with pore cement predominantly of smectite
composition, low values of filtration properties are due to the insignificant content of
large pore channels (less than 5%) and increasing number of non-filtered pores. These
formations were deposited in floodplain conditions. The class IV reservoirs include
fine- and medium-grained sandstones with polymineral cement: clayey pore and car-
bonate clot-pore type. Collectors of II–III classes are represented by coarse-grained and medium-grained
poorly sorted sandstones with pore-type cement, formed in an environment with a
relatively quiet hydrodynamic regime. 3
Results and Discussion At the first stage of creating a geological model of a terrigeneous reservoir, separate
strata were correlated. The selection and tracing of the layers was carried out by
logging diagrams; local surfaces associated with homogeneous rocks, more or less
sustained over the area, were taken as main reference points; within individual areas,
additional benchmarks were used, which were characterized by stable geophysical
characteristics. Two productive formations were confined to the Lower Triassic
deposits. To maximally take into account the features of the structure of productive
layers at the first stage of modeling, the study of microscopic in homogeneities of
productive sediments was carried out on the basis of lithofacies and sedimentation
models. The main reason for the heterogeneity of natural reservoirs in terms of the prop-
erties of the reservoirs and tires that form them is the conditions of their formation. Detailed lithofacies analysis of Lower Triassic deposits was carried out, based on a set
of investigations by both domestic (Muromtsev 1984; Bruzhes 2010; Morozov 2013)
and foreign researchers (Celli 1989; Hellem 1983). The analysis of geological and
geophysical information, the study of well cores, and the interpretation of well logging
data allowed reconstructing conditions for the formation of lower Triassic natural
reservoirs. The formation of these sediments took place in the continental conditions of
the alluvial plain. According to classification by A.G. Kossovskaya and M.I. Tuchkova (Kossovs-
kaya, Tuchkova 1988) sandstones fall into the field of polymictic (SiO2 content 62–
78%) and volcanictic (SiO2 content 54–64%). According to classification of Pettijohn
(1976) points of the composition of sandstones are localized in the fields of graywacke. 225 Oil and Gas Reservoirs in the Lower Triassic Deposits 4
Conclusions The analysis of the reservoir properties of sandstones shows that sandstones formed
under channel conditions are characterized by high median and average values of
porosity and permeability, respectively 24% and 20.8% (porosity), 56 10−12 and
40 10−12 m2, the lowest values characterize sediments formed in floodplain condi-
tions: the median values of porosity and permeability do not exceed 11% and
0.6 10−12 m2, arithmetic averages reach 14% and 1.6 10−12 m2. Analysis of the
graphs of the dependence of porosity-permeability for productive layers showed that
the highest values of filtration properties were typical to deposits of the basal layer,
where I–II class reservoirs were identified. A reliable seal of subregional distribution is the deposits of the Upper charkabo-
jskaya sub-suite, having high quality and thickness. Minerals of the smectite dominate
in the mineral composition, chlorite and illite are also present. Middle-Upper Triassic
clays are floodplain and lake originated, in their composition chlorite and illite pre-
dominate, which reduces the quality of the foamed seal compared to the Chark-
abozhskaya seal. 226 N. Timonina The differences in the composition and type of cement requires an individual
approach, a balanced choice of technologies in determining the development strategy
of fields and careful selection of a set of methods aimed at increasing oil recovery for
different sections of fields. Acknowledgements. The article was created under partial financial support by project of Ural
Branch of Russian Academy of Sciences №18-5-5-13 “The Geological Models, Environmental
Conditions and Prospects of Oil and Gas bearing of Phanerozoic deposits in Arctic regions of
Timan-Pechora province”. Timan-Pechora province”. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 227–230, 2019.
https://doi.org/10.1007/978-3-030-22974-0_55 Associated Petroleum Gas Flaring:
The Problem and Possible Solution A. Vorobev1,2(&) and E. Shchesnyak2 1 Atyrau University of Oil and Gas, Atyrau, Kazakhstan
fogel_al@mail.ru 2 Peoples’ Friendship University of Russia (RUDN University),
Moscow, Russia Moscow, Russia Abstract. The article analyzes the current state and prospects for utilization of
a hydrocarbon component dissolved in oil and released during its extraction and
preparation - associated petroleum gas (APG). The authors studied the proper-
ties, characteristics and component composition of APG. The analysis of the APG use at the international and regional levels is carried
out. The main causes of flaring were discussed and the shortage of production
capacities for APG processing in the Russian Federation was noted as one of the
main factors in the high level of APG flaring in the country. The paper notes possible ways of utilization of associated petroleum gas,
which depend on oil production conditions, such as field characteristics, oil/gas
ratio (gas-oil factor), and market opportunities for recovered gas. An overview
of all APG utilization methods are presented, which focuses on unit costs,
economic benefits and environmental impact reduction. The authors analyzed
the innovative experience of effective APG use in the USA and Canada. Special
attention is paid to the need to solve the problem of the effective use of APG in
the Russian Federation, especially the reduction of its burning in flare plants. Keywords: Associated petroleum gas Associated petroleum gas utilization
APG flaring Environmental pollution Associated Petroleum Gas Flaring:
The Problem and Possible Solution
A. Vorobev1,2(&) and E. Shchesnyak2
1 Atyrau University of Oil and Gas, Atyrau, Kazakhstan
fogel_al@mail.ru
2 Peoples’ Friendship University of Russia (RUDN University),
Moscow, Russia Associated Petroleum Gas Flaring:
The Problem and Possible Solution
A. Vorobev1,2(&) and E. Shchesnyak2
1 Atyrau University of Oil and Gas, Atyrau, Kazakhstan
fogel_al@mail.ru
2 Peoples’ Friendship University of Russia (RUDN University),
Moscow, Russia References Bruzhes LN, Izotov VG, Sitdikova LM (2010) Litofacial conditions of formation of the horizon
of the Yu1 Tevlinsko-Russkinskoye field of the West Siberian oil and gas province. Georesources 2(34):6–9 Celli RCh (1989) Ancient sedimentation. Nedra, Moscow, p 294 Celli RCh (1989) Ancient sedimentation. Nedra, Moscow, p 294 Hellem E (1983) Interpretation of facies and stratigraphic sequence. Mir, Moscow, p 328 Khanin AA (1976) Petrophysics of oil and gas reservoirs. Nedra, Moscow, p 259 Morakhovskaya ED (2000) Trias of the timan-ural region (reference sections, stratigraphy,
correlation). Biochronology and correlation of the Phanerozoic oil and gas basins of Russia,
vol 1, p 80. SPb: VNIGRI Morozov VP, Shmyrina VA (2013) Influence of secondary changes in reservoir rocks on
reservoir properties of the БC111 and ЮC11 productive layers of the Kustovoye deposit. Uchenye zapiski Kazan University, Kazan, vol 155, pp 95–98 Muromtsev VS (1984) Electrometric geology of sand bodies - lithological traps of oil and gas. Nedra, p 260 Pettijohn FJ, Potter PE, Siever R (1976) Sand and sandstone. Mir, Moscow, p 536 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction Associated petroleum gas (APG) is a kind of natural gas that is in oil deposits, either
dissolved in oil or as a free “gas cap” over oil into the deposits. Regardless of the
source, as soon as it separates from crude oil, it usually exists in mixtures of other
hydrocarbons such as ethane, propane, butane and pentane; in addition, APG contains
water vapor, hydrogen sulfide (H2S) and carbon dioxide (CO2), nitrogen (N2) and other
mixtures. Associated petroleum gas containing such impurities so it can not be trans-
ported and used without purification, as it is extracted in the process of oil production
(Kartamysheva et al. 2017). The volume and composition of APG depends on the area of production and on the
specific properties of the field. In the process of extraction and separation of one ton of
oil, it is possible to obtain from 25 to 800 m3 of associated gas. Some of this gas is used
or stored, because governments and oil companies have made significant investments A. Vorobev and E. Shchesnyak 228 for its extraction. However, individual companies burn APG because of technical,
regulatory or economic constraints. As a result, thousands of flare stacks from more
than 17,000 oil production facilities around the world burn about 140 billion cubic
meters of natural gas per year, resulting in more than 350 million tons of CO2 and a
large variety of pollutants, including very dangerous. The overall increase in global flaring compared to previous years is largely due to
negative developments in only a few countries: Iran, Russia and Iraq. Satellite data
show an increase in flaring in Iran by more than 4 billion m3, in Russia by almost 3
billion m3 and more than 1 billion m3 in Iraq. Flaring in Russia is close to the global
average compared to oil production; in the other two countries, flaring intensity is
higher. This is the expenditure of a valuable energy resource that can be used to
promote the sustainable development of producing countries. Thus, 149 billion m3 of
associated petroleum gas, burned in 2018, could turn into 750 billion kWh of elec-
tricity, which exceeds its total annual consumption by all countries of the African
continent. According to official data, the volume of extracted APG in Russia increased by
more than 7% - to 65 billion m3 in 2010 and over 70 billion m3 in 2018. 1
Introduction The immediate
impact on the increase in the volume of recoverable APG was made by the growth of
oil production in new areas, including the East Siberian fields. 2
Methods and Approaches For a long time, oil companies simply burned this unwanted by-product. Its flaring
requires a significant portion of the security system. The term “gas flaring” indicates a gas combustion (without energy recovery) in an
open flame, which is continuously lit on top of the flare stacks in the field of oil
production (Knizhnikov et al. 2017). Flaring occurs for three main reasons: • emergencies: limited incineration for reasons of safety for short periods of time can
always be necessary even after connecting the gas gathering pipeline; emergencies: limited incineration for reasons of safety for short periods of t
always be necessary even after connecting the gas gathering pipeline; • deficiency of gas utilization capacity - isolated well flaring: if the well starts to
produce oil and gas without connecting to gas gathering pipeline or other gas
utilization technology, the gas can be shut off; • lack of gas utilization capacity - well burning through the pipeline: if the well is
connected to gas gathering pipeline, but these systems can not process all gas from
the well (due to lack of power or compression), some or all of the associated gas
from the well can be flared. Billions of cubic meters of natural gas are burned at oil production sites around the
world. Gas combustion is a costly energy resource that can be used to support eco-
nomic growth and progress (Vorob’ev et al. 2017). Since 2012, the US National Oceanic and Atmospheric Administration and the
Global Gas Flaring Reduction Partnership have begun to apply a method for estimating
the volume of APG flared. This method consists in the use of satellite observational
data in the visible and near infrared ranges. 229 Associated Petroleum Gas Flaring Associated Petroleum Gas Flaring The conclusion drawn from the research results is that the volume of flared APG in
the world increased to 147 billion m3 in 2015, compared with 145 billion m3 in 2014
and 141 billion m3 in 2013. According to data for 2015, Russia led this “anti-rating”,
burning 24 billion m3 of APG, followed by Iraq (17.5 billion m3), Iran (16 billion m3)
and the United States (8 billion m3). Russia is also a “leader” (in third place after the
USA and Canada) with 1,814 flare stacks, which burn APG. At present, there are other possible ways of utilization of associated gas, alternative
to flaring. 2
Methods and Approaches Among them it is necessary to single out the following: • re-injection of APG into oil reservoirs to maintain pressure and increase oil
recovery (as a method of increasing oil recovery), or for possible conservation of it
as a resource and use in the future; • the use of gas as an energy source for production site or at oil producing facilities in
the vicinity; • the most effective way of utilization of associated petroleum gas is its processing at
gas processing plants to produce dry stripped gas (SOH), a wide fraction of light
hydrocarbons (LH), liquefied natural gas (LNG) and stable gasoline (SG). Below is an overview of all APG utilization methods, which focus on unit costs,
economic benefits and environmental impacts. Below is an overview of all APG utilization methods, which focus on unit costs,
economic benefits and environmental impacts. 3
Results and Discussion The indicator of its useful use has remained stable since the 2000s, within 73–79% of
the total amount of extracted APG in the country. Only in 2014–2017, according to the
public accounts of companies, it rose to 85–86%. According to the statement of rep-
resentatives of government organizations, the indicators of productive processing of
APG amounted to 90% in 2018. The amendments to the law “On environmental protection” (№219) adopted in
July 2014 caused such as significant reduction in the share of associated gas com-
bustion, according to these amendments, the company is obliged to establish its
technological standards at the level of application of the best available technologies. The total investment in increasing the useful use of APG was estimated at 200 billion
rubles. According to the Ministry of energy of the Russian Federation, it is expected
that the target of 95% of the associated gas will be used by the end of 2020 (Vorobyev
et al. 2018). Foreign experience of utilization of APG shows that flaring of gas in torches has
decreased slightly over the past two years, and oil production has also declined. In
particular, Nigeria reduced volume APG combustion to 8 billion m3 of nearly 18%
compared with 2013 year. The volume of associated gas flaring in the USA decreased
from 11 billion m3 in 2016 to less than 9 billion m3 in 2018 due to the use of a number
of innovative low-volume technologies. One of the innovative technologies for the production of liquefied natural gas with
small volumes of associated petroleum gas is LNG Production (Production Natural Gas
Liquids, LH-Pro). The “LH Pro” process combines dehydration, compression, cooling
and conditioning, eliminating the need for expensive glycol and refrigeration systems. 230 A. Vorobev and E. Shchesnyak Hydrate formation is excluded due to the thermal integration system. The technology
was developed by ASPEN and is used in the US and Canada. Hydrate formation is excluded due to the thermal integration system. The technology
was developed by ASPEN and is used in the US and Canada. 4
Conclusions Thus, the most rational ways of utilization of associated petroleum gas in Russia,
depending on the volumes of its extraction are: • at small volumes - covering own energy needs; • at small volumes - covering own energy needs; • with increased volumes - electricity generation and primary processing of APG to
produce lean dry gas (LDG) as fuel for the boiler room and light hydrocarbons
(LH) for disposal to the oil collector; • at resources from 50 to 150 million m3/year - processing with obtaining LDG, as
well as LH and electricity;
3 • with the amount of APG in excess of 150 million m3/year, processing of LDG, and
NGL is recommended. Knizhnikov AYU, Il’in AM (2017) Problems and prospects of associated petroleum gas use in
Russia. 2017 WWF Rossii, Moscow, p 34 Kartamysheva YeS, Ivanchenko DS (2017) Associated petroleum gas and the problem of its
utilization. Molodoy uchenyy. №25, pp 120–124 p
Vorob’ev A, Chekushina T, Vorob’ev K (2017) Russian national technological initiative in the
sphere of mineral resource usage. Rudarsko Geolosko Naftni Zbornik 2:1–8 References Kartamysheva YeS, Ivanchenko DS (2017) Associated petroleum gas and the problem of its
utilization. Molodoy uchenyy. №25, pp 120–124 Knizhnikov AYU, Il’in AM (2017) Problems and prospects of associated petroleum gas use in
Russia. 2017 WWF Rossii, Moscow, p 34 Vorob’ev A, Chekushina T, Vorob’ev K (2017) Russian national technological initiative in the
sphere of mineral resource usage. Rudarsko Geolosko Naftni Zbornik 2:1–8 Vorobyev AE, Zhang L (2018) Analysis of production and consumption of associated petroleum
gas in China. Bulletin of Atyrau Institute of oil and gas №2(46), pp 137–142 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction The development of oil and gas fields in complex mining and geological conditions is
associated with various complications - a drop in reservoir pressure, an increase in the
water content of the crude product and the amount of mechanical impurities,
destruction of the integrity of the well bottom zone. Analysis of the sources (Vorob’ev
et al. 2017) shows that sand manifestations are a multifactorial and multi-element
sophisticated technical system. The task of managing the sand formation processes includes such elements as sand
forecasting and effective methods of influencing the sand manifestation phases in order
to reduce negative effects. Innovative Technology of Using Anti-sand
Filters at Wells of the Vankor Oil
and Gas Field K. Vorobyev(&) and A. Gomes Department of Mineral Developing and Oil & Gas Engineering,
Engineering Academy, RUDN University, Moscow, Russia
k.vorobyev98@mail.ru Abstract. In this article, the authors considered an innovative technology for
restricting sands to reduce complications and watering during the development
of the Vankor gas and oil field. Keywords: Sand control filters Waterflood Well Borehole zone
Formation pressure drop Keywords: Sand control filters Waterflood Well Borehole zone
Formation pressure drop © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 231–232, 2019.
https://doi.org/10.1007/978-3-030-22974-0_56 References Lyashenko V, Vorob’ev A, Nebohin V, Vorob’ev K (2018) Improving the efficiency of blasting
operations in mines with the help of emulsion explosives. Min Mineral Deposits. 1:95–102
Ozhogina EG, Shadrunova IV, Chekushina TV (2017) Mineralogical rationale for solving
environmental problems of mining regions. Gornyi zhurnal. 11:105–110
Vorob’ev A, Chekushina T, Vorob’ev K (2017) Russian national technological initiative in the
sphere of mineral resource usage. Rudarsko Geolosko Naftni Zbornik. 2:1–8 Lyashenko V, Vorob’ev A, Nebohin V, Vorob’ev K (2018) Improving the efficiency of blasting
operations in mines with the help of emulsion explosives. Min Mineral Deposits. 1:95–102
Ozhogina EG, Shadrunova IV, Chekushina TV (2017) Mineralogical rationale for solving Lyashenko V, Vorob’ev A, Nebohin V, Vorob’ev K (2018) Improving the efficiency of blasting
operations in mines with the help of emulsion explosives. Min Mineral Deposits. 1:95–102 Lyashenko V, Vorob’ev A, Nebohin V, Vorob’ev K (2018) Improving the efficiency of blas
operations in mines with the help of emulsion explosives. Min Mineral Deposits. 1:95– operations in mines with the help of emulsion explosives. Min Mineral Deposits. 1:95 102
Ozhogina EG, Shadrunova IV, Chekushina TV (2017) Mineralogical rationale for solving
environmental problems of mining regions. Gornyi zhurnal. 11:105–110
Vorob’ev A, Chekushina T, Vorob’ev K (2017) Russian national technological initiative in the
sphere of mineral resource usage. Rudarsko Geolosko Naftni Zbornik. 2:1–8 p
p
p
p
Ozhogina EG, Shadrunova IV, Chekushina TV (2017) Mineralogical rationale for solving
environmental problems of mining regions. Gornyi zhurnal. 11:105–110
Vorob’ev A Chekushina T Vorob’ev K (2017) Russian national technological initiative in the Vorob’ev A, Chekushina T, Vorob’ev K (2017) Russian national technological initiative in the
sphere of mineral resource usage. Rudarsko Geolosko Naftni Zbornik. 2:1–8 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. 2
Methods and Approaches Studies confirm that during the removal of rock particles from the reservoir in the
operation of wells in the bottom zone form the high permeability channels of various
widths and lengths of fractures along and bedding planes, by which the bulk of gas and
produced water is filtered (Ozhogina et al. 2017). When considering the performance of geological and technical measures at the
stockwell of OJSC Gazprom (Lyashenko et al. 2018) in the direction of technology for
the elimination of sand production and the removal of wells from inactivity, the causes
and factors of sand production are given: poorly cemented formation; the viscosity of
the formation fluid; the velocity of fluid particles in the reservoir. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 231–232, 2019. https://doi.org/10.1007/978-3-030-22974-0_56 232 K. Vorobyev and A. Gomes 4
Conclusions According to the results of the analysis of the geological and technical conditions of
water wells and the characteristics of the aquifers of the Vankor oil and gas field, it was
decided to use downhole anti-sand filters to reduce the effect of removal of mechanical
impurities. The results of the pilot-scale industrial tests were considered successful,
which gave rise to the decision to implement filters on an industrial scale. As of January
2019, all water wells in the Vankor oil and gas field are equipped with anti-sand mesh
and slot-type filters 3
Results and Discussion Pilot tests of filters of various designs at the Vankor oil and gas field. During 2018, pilot
industrial tests offilters of various designs were carried out: slotted and multi-layer mesh. The filter descends on an unremovable packer into the perforated interval zone and
is placed opposite the entire interval in order to minimize the skin effect. The well is
operated using a high-rate installation of an electric submersible pump. At the base of a 168 mm column, filters with a total length of about 200 m are
lowered into the perforation intervals. Standard filter life is 24 months. Behind the
filter, another is formed - a natural filter. The organization of the reservoir pressure maintenance system at the Vankor oil
and gas field involves the use of both produced water and water from water wells. References If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Thermometry of Apatite Saturation
(The Kozhym Massif, The Subpolar Urals) Y. Denisova(&), A. Vikhot, O. Grakova, and N. Uljasheva Institute of Geology of the Komi SC UB of RAS,
Syktyvkar, Komi Republic, Russia
yulden777@yandex.ru Institute of Geology of the Komi SC UB of RAS,
Syktyvkar, Komi Republic, Russia
yulden777@yandex.ru Abstract. The results of the study of accessory apatite from the Kozhym massif
rocks have been presented in this paper. Apatites of the same morphological
type were found in granites. The Kozhym massif granites formation tempera-
tures by apatite were determined by the Watson and Bea saturation thermom-
etry. These temperatures were compared with the previously obtained ones for
accessory zircon of the same massif. Keywords: Apatite Granite The Kozhym massif The Subpolar Urals
Watson Bea Keywords: Apatite Granite The Kozhym massif The Subpolar Urals
Watson Bea 1
Introduction The Kozhym massif is located in the northeastern part of the Subpolar Urals on the left
and right banks of the Kozhym River in the Oseu and Ponyu Rivers basins (Fig. 1)
among deposits of the Puyvinian Middle Riphean Formation. This massif is the second
by area among the geobodies composing the Kozhym intrusion which includes the
Kuzpuayu granite massif. The most fully preserved Kozhym massif granites are
medium-grained pink greenish-gray rocks. They (the rocks) are characterized by a
massive coarse-platy texture with well-defined tectonic gneissiness. The studied massif
belongs to the A-type according to B. Chappel classification (Fishman et al. 1968). The accessory massif apatite is represented by yellow mat elongated crystals of a
hexagonal dipyramidal-prismatic habitus. The crystals size is 0.1–0.4 mm, the elon-
gation coefficient is 1.5–3. The mineral shape is represented by a combination of a
prism (1010) and a dipyramid (1011). The faces surface is fractured. Characteristic
inclusions are quartz, plagioclase, zircon. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 235–238, 2019.
https://doi.org/10.1007/978-3-030-22974-0_57 2
Methods 1992) proposed the following addition to the Watson’s formula for
peraluminum rocks (A/CNK > 1): P2O5 Bea
ð
Þ ¼ P2O5 HW
ð
Þ P2O5 HW
ð
Þ e
6429 A=CNK1
ð
Þ
T273;15
: 2
Methods Apatite is increasingly used as a geothermometer. The E. Watson saturation ther-
mometry was used to determine the mineral crystallization temperatures. This method
allowed determining the apatite and rock formation temperature by the distribution of
the phosphorus oxide content between apatite and the rock containing the mineral. The
level of phosphorus saturation necessary for the apatite crystallization depends on the © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 235–238, 2019. https://doi.org/10.1007/978-3-030-22974-0_57 236 Y. Denisova et al. Fig. 1. The geological Kozhym granite massif map. 1 – mica-quartz schists, green orthoschists,
quartzits; 2 – mica-quartz schists, porphyries, porphyrites, marbles and quartzits interlayers; 3 –
granites; 4 – gabbro; 5 – contact lines: a – stratigraphic and magmatic, b – tectonic; 6 – planar
structured bedding elements. Massifs (numbers in circles): 1 – the Kuzpuayu; 2 – the Kozhym. Fig. 1. The geological Kozhym granite massif map. 1 – mica-quartz schists, green orthosch
quartzits; 2 – mica-quartz schists, porphyries, porphyrites, marbles and quartzits interlayers;
granites; 4 – gabbro; 5 – contact lines: a – stratigraphic and magmatic, b – tectonic; 6 – pla
structured bedding elements. Massifs (numbers in circles): 1 – the Kuzpuayu; 2 – the Kozh Fig. 1. The geological Kozhym granite massif map. 1 – mica-quartz schists, green orthoschists,
quartzits; 2 – mica-quartz schists, porphyries, porphyrites, marbles and quartzits interlayers; 3 –
granites; 4 – gabbro; 5 – contact lines: a – stratigraphic and magmatic, b – tectonic; 6 – planar
structured bedding elements. Massifs (numbers in circles): 1 – the Kuzpuayu; 2 – the Kozhym. silica content in the meta-aluminic rock (A/CNK < 1) and the temperature according to
the Harrison and Watson calculations (Harrison et al. 1984): InDp ¼ 8400 þ 26400 SiO2 0; 5
ð
Þ
ð
Þ=T 3; 1 12; 4 SiO2 0; 5
ð
Þ;
P2O5 HW
ð
Þ ¼ 42=Dp, where Dp is the ratio of P concentration in apatite and melt, P2O5; SiO2 is the weight
fraction of the phosphorus oxide, silica in the melt, wt. %, T is the temperature, Kelvin. where Dp is the ratio of P concentration in apatite and melt, P2O5; SiO2 is the weight
fraction of the phosphorus oxide, silica in the melt, wt. %, T is the temperature, Kelvin. Bea (Bea et al. Thermometry of Apatite Saturation (The Kozhym Massif, The Subpolar Urals) Thermometry of Apatite Saturation (The Kozhym Massif, The Subpolar Urals) 237 Table 1. Chemical granites composition of the Kozhym massif Table 1. Chemical granites composition of the Kozhym massif
Component, wt. % Sample number
К-1
К-2
К-3
К-4
К-5
К-6
К-7
К-8
К-9
К-10
SiO2
77.78
76.89
75.95
75.89 76.49
78.12
77.54
76.26 77.48 76.95
TiO2
0.16
0.22
0.48
0.52
0.48
0.11
0.24
0.42
0.31
0.59
Al2O3
11.88
11.95
12.69
12.52 10.05
11.34
11.78
12.22 11.09 10.92
FeO
1.72
1.29
1.15
1.24
0.56
0.50
0.59
0.62
0.61
1.03
Fe2O3
0.84
1.12
0.52
1.05
0.92
1.21
0.87
1.02
0.89
0.56
MnO
0.02
0.00
0.00
0.01
0.02
0.01
0.02
0.04
0.03
0.03
MgO
0.16
0.25
0.17
0.33
0.38
0.39
0.18
0.29
0.19
0.18
CaO
0.31
0.29
0.22
0.38
0.59
0.28
0.45
0.42
0.37
0.51
Na2O
3.65
3.33
4.22
4.02
3.08
3.15
3.22
4.51
3.01
3.89
K2O
3.88
4.51
4.09
3.89
4.15
5.17
4.99
3.78
4.65
3.28
P2O5
0.01
0.03
0.02
0.02
0.03
0.02
0.01
0.01
0.03
0.02
ппп
0.05
0.29
0.59
0.15
0.75
0.62
0.39
0.98
1.02
1.23
P
100.46 100.17 100.10 100.02 97.50 100.92 100.28 100.57 99.68 99.19
Note. The chemical composition was obtained using the silicate method in CUC Science of Institute of
geology of Komi SC UB RAS (analyst Koksharova O.V.) Note. The chemical composition was obtained using the silicate method in CUC Science of Institute of
geology of Komi SC UB RAS (analyst Koksharova O.V.) Table 2. Saturation temperatures for the Kozhymmassif apatite
Temperature, °C
Sample number
К-1 К-2 К-3 К-4 К-5 К-6 К-7 К-8 К-9 К-10
According to Watson 784 860 819 818 856 838 782 770 865 828
According to Bea
722 798 770 764 856 836 768 770 840 826
Fig. 2. Formation temperatures distribution histograms for the Kozhym massif granites. a - according to Watson, b - according to Bea Table 2. Saturation temperatures for the Kozhymmassif apatite
Temperature, °C
Sample number
К-1 К-2 К-3 К-4 К-5 К-6 К-7 К-8 К-9 К-10
According to Watson 784 860 819 818 856 838 782 770 865 828
According to Bea
722 798 770 764 856 836 768 770 840 826
Fig. 2. Formation temperatures distribution histograms for the Kozhym massif granites. a - according to Watson, b - according to Bea Table 2. Saturation temperatures for the Kozhymmassif apatite Fig. 2. Formation temperatures distribution histograms for the Kozhym massif granites. 3
Results and Discussion The apatite saturation temperatures based on the data of the chemical granites com-
position of the Kozhym massif (Table 1) were calculated by the author according to
Watson and Bea (Table 2). The Kozhym massif rocks formation temperatures distribution histograms by
apatite were compiled on the obtained temperatures for each calculation method
(Fig. 2). Thermometry of Apatite Saturation (The Kozhym Massif, The Subpolar Urals) a - according to Watson, b - according to Bea The presented histograms show that the studied granites are high-grade rocks. The
Kozhym massif formation occurred at temperatures from 770 °C to 865 °C and
averaged 822 °C according to Watson. The application of Bea refinements allowed
concluding that the massif rocks formation occurred at temperatures from 722 °C to
856 °C and an average of 795 °C. 238 Y. Denisova et al. 4
Conclusions The Kozhym massif rocks formation occurred at high temperatures. The similar massif
granites formation temperature ranges: 770–865 °C according to Watson, 722–856 °C
according to Bea were obtained using the Watson saturation thermometry by apatite
and the Watson formula adjustment for aluminous rocks according to Bea. The data
confirmed the temperatures obtained earlier by the author using the Watson saturation
thermometry (749–816 °C) and the classical evolutionary-morphological Pupin anal-
ysis (700–900 °C) for zircon of the same massif (Denisova 2016; 2018a; b). It can be
affirmed that the Watson saturation thermometry and the refined Bea formula for apatite
provide the same information about regime temperature evolution during the granites
formation such as on the Watson saturation thermometry and the Pupin and Tyurko
evolutionary-crystallomorphological analysis for zircons. Acknowledgements. The work was supported by the Basic Research Program of RAS
№18-5-5-19. Acknowledgements. The work was supported by the Basic Research Program of RAS
№18-5-5-19. References Bea F, Fershtater GB, Corretgé LG (1992) The geochemistry of phosphorus in granite rocks and
the effects of aluminium. Lithos 48:43–56 Bea F, Fershtater GB, Corretgé LG (1992) The geochemistry of phosphorus in granite rocks and
the effects of aluminium. Lithos 48:43–56 Bea F, Fershtater GB, Corretgé LG (1992) The geochemistry of phosphorus in granite rocks and
the effects of aluminium. Lithos 48:43–56 Denisova UV (2018a) Apatite of the Nikolaishor granite massif (the Subpolar Urals). Vestnik of
Institute of geology of Komi SC UB RAS, Syktyvkar, №9, pp 24–29 Denisova UV (2018b) Crystal morphology of zircon in solving problems of the Kozhimsky
massif granites genesis (the Subpolar Urals). Trends in the development of science and
education, №35, Part 3. Samara, pp 45–48. https://doi.org/10.18411/lj-28-02-2018-51 Denisova UV (2016) Temperature survey of zircon from the granitoids of the Subpolar Urals. Vestnikof Institute of geology of Komi SC UB RAS, Syktyvkar, №12, pp 37–44 Denisova UV (2016) Temperature survey of zircon from the granitoids of the Subpolar Urals. Vestnikof Institute of geology of Komi SC UB RAS, Syktyvkar, №12, pp 37–44 Fishman MV, Yushkin NP, Goldin BA, Kalinin EP (1968) Mineralogy, typomorphism and
genesis of accessory igneous rocks minerals of the Urals and Timan north. M.-L.: Sci 252 Fishman MV, Yushkin NP, Goldin BA, Kalinin EP (1968) Mineralogy, typomorphism and
genesis of accessory igneous rocks minerals of the Urals and Timan north. M.-L.: Sci 252 Harrison TM, Watson EB (1984) The behavior of apatite during crustal anatexis: equilibrium and
kinetic considerations. Geochim Cosmochim Acta 48:1467–1477 Harrison TM, Watson EB (1984) The behavior of apatite during crustal anatexis: equilibrium and
kinetic considerations. Geochim Cosmochim Acta 48:1467–1477 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Studies of Structural Changes in Surface
and Deep Layers in Magnetite Crystals
After High Pressure Pressing P. Matyukhin(&) Belgorod State Technical University named after V.G. Shukhov,
Belgorod, Russia
mpvbgtu@mail.ru Abstract. The article introduces the study of structural changes in surface and
deep layers in magnetite crystals, on samples under high pressure pressing. Magnetite (magnetite iron-ore concentrate) is widely used as a filling compound
of new composites which are planned to be used in nuclear-construction. These
compounds are based on the aluminum containing matrix with a filling com-
pound. This modern composite material can be used in construction structures
able to resist significant loads, operate in such extreme situations as abrupt
dynamic loads, fires with further alternating temperature oscillations. Keywords: Magnetite Pressure Crystal Structure Layer Material Keywords: Magnetite Pressure Crystal Structure Layer Material © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 239–243, 2019.
https://doi.org/10.1007/978-3-030-22974-0_58 2
Methods and Approaches As an object of research we used a highly-dispersive enriched magnetite iron-ore
concentrate from the Lebedinsk deposit of the Kursk Magnetic Anomaly with density
4950 kg/m3, Mohs hardness – 6, black colour; the magnetite is in form of irregular
needle-shaped grains, octahedral crystals, shell-like fracture; the mineral composition is
introduced by magnetite with inclusions of carbonate (chalk-stone) and siliceous
(quartz) admixtures. After cleaning and chemical treatment of the magnetite iron ore
concentrate per 99,8% its mineral composition is introduced by magnetite (FeO.Fe2O3)
with fractional composition of particles 0,05–15 mcm. In the research we used a
modern electronic microscope “TESCAN MIRA 3 LMU”. 1
Introduction Nowadays many scientists develop new kinds of materials, including composites which
can be used in nearly all spheres of human life. Composite materials based on organic
and inorganic components with different additives and fillers are created. These
components can be the base for different matrixes. Fillers with different qualities are
introduced into these matrixes depending on the purpose, scope of operation, condi-
tions of service of the designed composite. These matrixes can have metal, ceramics,
concrete, polymer and other bases, and the fillers may be cast iron shot, barites, metal
processing remains, mining company products (for example, iron containing rocks) and
of other enterprises (Grishina and Korolev 2016; Barabash et al. 2017; Matyukhin
2018; Gulbin et al. 2018; Kruglova 2009; Laptev et al. 2015; Gulbin et al. 2016;
Matyukhin et al. 2011, Samoshin et al. 2017). Composite materials based on metal matrixes can be used in bearing constructions
production. Such constructions can resist high mechanical impacts, intensive ionizing
radiation and alternating temperatures, that can widely be used in nuclear construction
industry, and foremost, for biological protection on sites with power supplies of dif-
ferent nature. Nowadays, the issue of aluminum containing matrixes and iron con-
taining fillers utilization in designing new types of radiation-protective construction
composite materials becomes acute. One of promising fillers for new composite © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 239–243, 2019. https://doi.org/10.1007/978-3-030-22974-0_58 P. Matyukhin 240 materials is magnetite. The introduction of modern composite materials of natural iron
ore raw materials will give them additional mechanical strength, increase their radiation
shielding characteristics. This filler is widely found in nature, is relatively cheap, and
composites on its base will meet ecological requirements as they are a part of the
ecosystem. Here a possibility of combining magnetite iron ore concentrate with alu-
minum containing matrix and receiving composite material by high pressure pressing is
of great scientific interest. In this research it is necessary to solve issues about maxi-
mum possible filler compacting in its composite material matrix, studying composite
filler behaviour at high pressing pressures, in particular studying structural changes in
surface and deep layers of magnetite crystals at pressing pressures when such a
composite material is produced, and compact-space position of its particles relative to
each other. 3
Results and Discussion For experiment fairness the magnetite samples in the form of a high dispersive powder
with particles 0,05–15 mcm were used, without adding it into the aluminium con-
taining matrix. Figure 1 shows the structure of surface and deep layers of magnetite
particles without pressure treatment (Fig. 1a) and after pressure pressing 5000 MPa
(Fig. 1b) with magnifying power 10 mcm. The analysis of the received data showed
that the surface and deep layers of magnetite crystal particles after pressure pressing at
5000 MPa (Fig. 1b), in comparison with the micro photo of magnetite crystal particles
not pressed (Fig. 1a), there are spaces with loose distribution of its particles along the
whole sample volume: there are numerous empty spaces; particles have irregular form
with hard aggregation and rough edges. Figure 2 shows the structure of surface and
deep magnetite particle layers, after pressure 10000 MPa and 20000 MPa with mag-
nifying power 10 mcm. 241 Studies of Structural Changes in Surface and Deep Layers Fig. 1. Magnetite surface with magnifying power 10 mcm: (a) without pressure pressing
treatment, (b) after pressure treatment 5000 MPa Fig. 1. Magnetite surface with magnifying power 10 mcm: (a) without pressure pressing
treatment, (b) after pressure treatment 5000 MPa The received data analysis showed that at pressure pressing increase up to
10000 MPa (Fig. 2a) magnetite crystal particles were distributed more compactly
within the whole volume of the studied sample: also there are numerous empty spaces
but with a slight reduction of their geometrical sizes; the magnetite particles structure
changed insignificantly, as before they had irregular form with strong aggregation and
rough edges, but here particles of smaller fraction appeared, that evidenced the process
of partial mechanical destruction of its particles. Fig. 2. Magnetite surface with magnifying power 10 mcm: (a) after pressure pressing
10000 MPa, (b) after pressure pressing 20000 MPa Fig. 2. Magnetite surface with magnifying power 10 mcm: (a) after pressure pressing
10000 MPa, (b) after pressure pressing 20000 MPa When the pressure increases up to 20000 MPa on the magnetite crystal particles
(Fig. 2b), there is a more compact visual magnetite particle packing both in surface and
deep layers of the studied sample; there is a significant increase of zones with good
compaction. 4
Conclusions Based on the studies of structure changes in surface and deep layers of magnetite
crystals, after high pressure pressing and studied with electron microscope, we can make
an assumption that the structure of the composite may be denser by adding magnetite
into its filler 15–25% (by volume) of its particles, but smaller fraction in comparison the
studied one. It is theoretically possible that after adding such an amount of magnetite
fine-fraction a higher degree of sample material compacting will be achieved. Or to
conduct the studies with adding the same percentage ratio into the volume of studied
composite material matrix samples in the form of aluminum powder, as it has particles
less in size than magnetite particles. We think that these studies have high theoretical
and practical significance and they should be taken into account when developing new
composite materials based on magnetite filler and different metal matrixes. Acknowledgements. The work is realized in the framework of the Program offlagship university
developmentonthebaseoftheBelgorodStateTechnologicalUniversitynamedafterV.G.Shukhov,
using equipment of High Technology Center at BSTU named after V.G. Shukhov. Acknowledgements. The work is realized in the framework of the Program offlagship university
developmentonthebaseoftheBelgorodStateTechnologicalUniversitynamedafterV.G.Shukhov,
using equipment of High Technology Center at BSTU named after V.G. Shukhov. 3
Results and Discussion There is a greater degree of destruction of magnetite crystal particles
surface in the total amount of material samples in comparison with previous pressure
pressing; somewhere surface aggregation and hematite particles edges is smoothed. P. Matyukhin 242 It evidences qualitative distribution of hematite particles of all fraction composition along
the whole sample volume, but micro photos show remaining zones with “insufficient
compaction”, where there is a lack of fine-fractioned magnetite particles. It evidences qualitative distribution of hematite particles of all fraction composition along
the whole sample volume, but micro photos show remaining zones with “insufficient
compaction”, where there is a lack of fine-fractioned magnetite particles. Studies of Structural Changes in Surface and Deep Layers Ochkina NA (2018) Heat stability of radio-protective composite based on aluminous cement and
polymineral industrial waste. Pridneprov Sci Bull 3(2):007–010 Samoshin AP, Korolev YV, Samoshina YN (2017) Internal stresses at metal concrete structure
formation for protection from radiation. Bull. BSTU Named After V.G. Shukhov 6:13–17 Ochkina NA (2018) Heat stability of radio-protective composite based on aluminous cement and
polymineral industrial waste. Pridneprov Sci Bull 3(2):007–010
Samoshin AP, Korolev YV, Samoshina YN (2017) Internal stresses at metal concrete structure
formation for protection from radiation. Bull. BSTU Named After V.G. Shukhov 6:13–17 References Barabash DE, Barabash AD, Potapov YuB, Panfilov DV, Perekalskiy OE (2017) Radiation-
resistant composite for biological shield of personnel. In: IOP conference series: earth and
environmental science. C. 012085 Grishina AN, Korolev EV (2016) New radiation-protective binder for special-purpose
composites. Key Eng Mater 683:318–324 Gulbin VN, Kolpakov NS, Gorkavenko VV, Boikov AA (2018) Research of the structure and
properties of radio and radiation-protective polymer nanocomposites. J Electro-magnetic
Waves Electron Syst 23(1):4–11 Gulbin VN, Martsenuk AV, Gorkavenko VV, Cherdyntsev VV (2016) Polymeric composites for
radio and radio active protection. Sci Intensive Technol 17(10):7–12 Kruglova AN (2009) Radiation protective materials based on industrial wastes: physic-
mechanical properties. Reg Archit Const 1:53–56 Laptev GA, Potapov Y, Yerofeev VT (2015) Development of manufacturing technology
metalloconcretes. Build Reconst 1(57):123–129 Matyukhin PV (2018) The choice of iron-containing filling for composite radioprotective
material. In: IOP conference series: materials science and engineering 11. International
conference on mechanical engineering, automation and control systems 2017 - material
science in mechanical engineering. C. 032036 Matyukhin PV, Pavlenko VI, Yastrebinskiy RN, Bondarenko YuM, (2011) Prospects of creating
modern highly constructive radiation-protective metalocomposites. Bulletin of BSTU named
after V.G. Shukhov. 2, 97 Studies of Structural Changes in Surface and Deep Layers 243 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. The Potential of Lacquer Peel Profiles
and Hyperspectral Analysis for Exploration
of Tailings Deposits W. Nikonow(&) and D. Rammlmair Federal Institute for Geosciences and Natural Resources (BGR),
Hanover, Germany
wilhelm.nikonow@bgr.de Abstract. Lacquer peel profiles are a valuable technique to preserve and study
sedimentary structures and depositional features outside the field. The use of the
lacquer fluid Mowiol® has shown to work well as a simple and rapid prepa-
ration technique for tailings material from a tailings heap in Copiapó, Chile. The
combination of lacquer peels with 2D mapping techniques such as Hyper-
spectral Imaging (HSI) can provide important sedimentary information such as
particle size distribution on a large scale with little effort, which becomes
quantifiable due to continuous 2D information and modern image analysis. The
presented relationship of element or mineral distribution with particle size can
serve as a tool for targeted and more focused sampling for mineral exploration in
tailings and tailings analysis in general, considering future selective mining for
economic or environmental reasons. Keywords: Lacquer profile Sedimentology Hyperspectral Imaging
Tailings exploration Keywords: Lacquer profile Sedimentology Hyperspectral Imaging
Tailings exploration © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 244–247, 2019.
https://doi.org/10.1007/978-3-030-22974-0_59 2
Methods and Approaches The lacquer profiles were taken from a tailings heap in the city of Copiapó in northern
Chile. According to historic aerial images it was deposited ca. 1993 on an area of ca. 230 130 m with an approximated mass of 400.000 t in 2018. There is only little
information about the origin of the heap, but the material is most probably from a
nearby processing plant using flotation or leaching to process ore from one of the many
mines in the Punta del Cobre district around Copiapó. Since the heap is not covered, it
is a target for erosion by wind and rain affecting the adjacent area including the
Copiapó River. Three lacquer profiles were taken from the steep parts of the eastern and western
sides, where the internal layering becomes visible and accessible. To create the profiles
the polyvinyl alcohol Mowiol® 4-88 was used. It is dissolved in hot water a day before
preparation. At the site, the selected surface is scraped even and the Mowiol is applied
on the surface by a spray bottle or a brush. For stabilization, several layers of Mowiol
and gauze are laid on the back of the profile. Then the profiles are left to dry for about
24 h. Finally, they can be removed with a knife and, wrapped in cloth, can be trans-
ported easily. Preparation of one profile of about 30 50 cm takes about 30 min
excluding the time for drying. One profile was taken from the eastern side (LP1) and
two profiles from the western side (LP2, LP3). Additional samples from each layer
were taken next to the profiles for geochemical, particle size analysis and verification of
the HSI analyses. The dry profiles were analyzed in the laboratory with the SisuRock system from
Specim. The images were taken with two push broom cameras covering the wavelength
range from 400 to 2500 nm (VNIR and SWIR) at a spatial resolution of 875 lm
(SWIR) and 280 lm (VNIR). The data were processed using vertical de-striping and
minimum noise fraction transform. 1
Introduction Creation of lacquer peels is an old, but rarely recognized technique enabling preser-
vation of sedimentary structures of granular materials. The profiles are also known as
sedimentary peels, sediment plates or lacquer profiles and are similar to the early
description of soil monoliths by e.g. Voigt (1936). As a relatively simple and inex-
pensive technique with great possibilities in various geoscientific fields it represents a
valuable method to study and preserve complex depositional processes and sedimen-
tary textures on a scale from millimeters up to several square meters. The general procedure is to apply the lacquer fluid on the target area, then let the
fluid dry, solidify, remove the peel and fixate it on a board for transport and display. A detailed technical description of the preparation process can be found e.g. in Van
Baren and Bomer (1979). The impregnation takes place due to gravity and capillary
forces, which makes it possible to create vertical profiles. The advantage of lacquer peel preparation is the possibility to preserve and
transport features of interest for additional analysis on the site or in the laboratory. The
combination of lacquer peels with emerging analytical techniques for 2D mapping such 245 The Potential of Lacquer Peel Profiles and Hyperspectral Analysis as Hyperspectral Imaging (HSI) can provide non-destructively valuable information
from e.g. textural, sedimentological and geochemical analysis. HSI is commonly used in remote sensing and finds its path into other fields of
geoscience. HSI measures the reflectance of the samples in a wavelength range of, in
this work, the visible and near infrared region (VIS-NIR: 400–2500 nm). It utilizes
molecule - light interaction such as vibrational processes or electron energy level
transitions which produce characteristic absorption features at certain wavelengths
(Hunt and Salisbury 1970). 3
Results and Discussion From the tailings impoundment three lacquer profiles were prepared. The profiles were
left to dry for about 24 h to minimize the risk of destroying structures during removal
and transport. When the profiles were removed from the tailings wall, only small areas
at the edges or in clay layers were lost, but overall, the texture was well preserved. 246 W. Nikonow and D. Rammlmair Hyperspectral images of all profiles were obtained in the laboratory with the
SisuRock system within minutes. The data show clearly the layered structure, which
seems to be mostly an effect of particle size. Figure 1 shows the optical and the
hyperspectral image of LP2. Layers that were differentiated optically in the field are
indicated by capital letters. In the hyperspectral image areas of coarse particle size
appear dark, while layers of fine particle size appear light grey or white. The data show
a regression coefficient of R2 = 0.76 at 860 nm between particle size and reflectance. Fig. 1. Left: True color photo of the lacquer peel LP2 with layers selected for bulk sampling. Middle: Hyperspectral scan of LP2 at 860 nm. The red line indicates the position of the
reflectance profile in the right graph. Right: Distribution of reflectance (red line) at 860 nm and
average particle size (black line) at lacquer peel LP2. Fig. 1. Left: True color photo of the lacquer peel LP2 with layers selected for bulk sampling. Middle: Hyperspectral scan of LP2 at 860 nm. The red line indicates the position of the
reflectance profile in the right graph. Right: Distribution of reflectance (red line) at 860 nm and
average particle size (black line) at lacquer peel LP2. The effect of particle size on reflectance in the visible and infrared wavelength
region has been studied in the literature and was described as a negative correlation
(Hunt and Vincent 1968; Okin and Painter 2004; Salisbury and Hunt 1968). However,
the application on and implications for tailings evaluation still have to be described. The particle size distribution is an important information for the evaluation of the
economic value of tailings deposits. The extractability of e.g. Cu depends largely on the
particle size of the Cu minerals, since flotation or heap leaching depend on the access to
free mineral surfaces. 4
Conclusions The proposed method of creating lacquer profiles has worked well on tailings material. It is a simple and rapid method to preserve sedimentary features for display and further
analysis. The combination with 2D mapping techniques such as HSI provides rapidly
continuous 2D information on sedimentary structures and particle size distribution on a
scale from micrometer to meter. The information is quantifiable by image analysis and
can be used for various geoscientific applications including sedimentology, sequential
analysis and tailings exploration. Acknowledgements. The results of this work are part of research that is funded by the German
Federal Ministry of Education and Research (BMBF) within the projects SecMinStratEl
(033R118B) and SecMinTec (033R186B). We thank Dominic Göricke for preparation of the
lacquer profiles and Dr. Martin C. Schodlok for support with the hyperspectral data processing. We are very grateful to the reviewers for their helpful comments. 3
Results and Discussion The geochemical data from the additional samples analyzed by
XRF show a possible trend of increasing Cu concentration with increasing particle size
above 100 lm. These areas of interesting particle size can be identified, located and
quantified with HSI. After establishing the methods in the laboratory, it could be
expanded to drill cores or in-situ measurements in the field. Therefore, the combination
of creating lacquer profiles and HSI and image analysis serves as a rapid and simple
tool to acquire important information for targeted and more selective sampling and
even evaluation of tailings material for possible selective mining. 247 The Potential of Lacquer Peel Profiles and Hyperspectral Analysis Methods of Extraction of Micro-
and Nanoparticles of Metal Compounds
from Fine Fractions of Rocks, Ores
and Processing Products Abstract. The presented extraction methods are based on the features of the
state of liquids in the capillary space in the form of a weak electrolyte. These
methods make possible to extract micro- and nanoparticles adsorbed in matrix
minerals from a suspension placed in the graphite substrate into a capillary
solution. After the particle deposition in the substrate due to evaporative con-
centration the microprobe analysis is performed. The method is known as the
capillary method of extracting micro and nanoparticles. The described methods was used as a prototype for extracting micro- and
nanoparticles from suspensions associated with the use of an external electric
field. The field is created by connecting the electrodes to a graphite substrate and
applied suspension with a direct current source using the voltage of 4 6 V. The micro- and nanoparticles adsorbed in matrix minerals are extracted into the
capillary solution. The deposition of micro- and nanoparticles in a capillary
solution is made by the method of evaporative concentration. The application of
an external electric field intensify extraction of micro- and nanoparticles. Keywords: Nanoparticles Capillaries Electrolyte Graphite
Electric potential Adsorption Keywords: Nanoparticles Capillaries Electrolyte Graphite
Electric potential Adsorption References Hunt GR, Salisbury JW (1970) Visible and near-infrared spectra of minerals and rocks: I silicate
minerals. Mod Geol 1:283–300 Hunt GR, Vincent RK (1968) The behavior of spectral features in the infrared emission from
particulate surfaces of various grain sizes. J Geophys Res 73:6039–6046 Hunt GR, Vincent RK (1968) The behavior of spectral features in the infrared emission from
particulate surfaces of various grain sizes. J Geophys Res 73:6039–6046 Okin GS, Painter TH (2004) Effect of grain size on remotely sensed spectral reflectance of sandy
desert surfaces. Remote Sens Environ 89:272–280 Okin GS, Painter TH (2004) Effect of grain size on remotely sensed spectral reflectance of sandy
desert surfaces. Remote Sens Environ 89:272–280 Salisbury JW, Hunt GR (1968) Martian surface materials: effect of particle size on spectral
behavior. Science 161:365–366 Salisbury JW, Hunt GR (1968) Martian surface materials: effect of particle size on spectral
behavior. Science 161:365–366 Van Baren J, Bomer W (1979) Procedures for the collection and preservation of soil profiles. International soil museum Van Baren J, Bomer W (1979) Procedures for the collection and preservation of soil profiles. International soil museum Voigt E (1936) Die Lackfilmmethode, ihre Bedeutung und Anwendung in der Paläontologie,
Sedimentpetrographie und Bodenkunde. Zeitschrift der deutschen geologischen Gesellschaft
88:272–292 Voigt E (1936) Die Lackfilmmethode, ihre Bedeutung und Anwendung in der Paläontologie,
Sedimentpetrographie und Bodenkunde. Zeitschrift der deutschen geologischen Gesellschaft
88:272–292 Voigt E (1936) Die Lackfilmmethode, ihre Bedeutung und Anwendung in der Paläontologie,
Sedimentpetrographie und Bodenkunde. Zeitschrift der deutschen geologischen Gesellschaft
88:272–292 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 248–251, 2019.
https://doi.org/10.1007/978-3-030-22974-0_60 1
Introduction A - clean graphite plate; B - a plate with a dried hemisphere of the investigated
suspension; C- a plate with a hemisphere removed; D - Dried hemisphere (flat side up) 1
Introduction The following methods of extraction of minerals trace from rocks, ores and products of
their processing are well known. 1. In heavy liquids with subsequent magnetic and electromagnetic separation of heavy
fractions (Chueva 1950; Mitrofanov et al. 1974). The disadvantage of this method is
the aggregation of nanoparticles with minerals with a particle size less than
0.045 mm and their absence in the heavy fraction. 2. Extraction of rhenium from ores of black shale formations (Oleynikova et al. 2012. The method for the extraction of micro- and nanoparticles of metal compounds is
not calculated. 3. Extraction of nanoparticles from disperse systems (Zhabreev 1997) using the
electric field created by electromagnets. The method does not provide for the
extraction of all classes of metal compounds. 3. Extraction of nanoparticles from disperse systems (Zhabreev 1997) using the
electric field created by electromagnets. The method does not provide for the
extraction of all classes of metal compounds. 249 Methods of Extraction of Micro- and Nanoparticles of Metal Compounds 4. Method of capillary extraction of nanoparticles (Smetannikov 2014). The material
of fine fractions (<0.25 mm) mixing with a liquid forms of suspension, placed in the
form of a hemisphere in a graphite substrate. The distance between minerals is
comparable to their size and forms a capillary space. The liquid acquires structure,
charge and becomes electrolyte (Deryagin et al. 1989). Nanoparticles adsorbed in
matrix minerals are extracted into the capillary solution. Due to the evaporation concentration, the nanoparticles migrate to the base of the
hemisphere. After drying (1 1.5 h), the hemisphere is removed, and the nanoparti-
cles on the substrate are examined under a microprobe (Fig. 1). More than 50 minerals
have been identified: intermetallic compounds and solid solutions of Au, Cu, Ag, Zn,
Pb, Ni, Sn, Cu, Fe, Cr, Ti zircon, Sn minerals, monazite, which are trace substances in
the water-insoluble salt residues of the Verkhnekamsk deposit (Smetannikov and
Filippov 2010). The disadvantage of this method is incomplete extraction of nanoparticles. This
method served as a prototype for developing method for extracting particles using the
methods of intensifying the natural properties of capillary systems by an external
electric field. Fig. 1. A - clean graphite plate; B - a plate with a dried hemisphere of the investigated
suspension; C- a plate with a hemisphere removed; D - Dried hemisphere (flat side up) Fig. 1. 2
Methods and Approaches The suspension is prepared from the material of fine fractions using distilled water and
adding salt. The suspension is applied to a graphite plate, forming the hemisphere. An
electrode with a “plus” sign (forming an anodic part of the system with a graphite plate)
is connected to the underside of the plate. The second electrode is immersed in the
suspension (without touching the graphite substrate) with a minus sign serves as a
cathode. Then there is inclusion of the direct current voltage of 4 6 V. Further there
is an electrolytic extraction of nanoparticles and deposition on the substrate. The dried
hemisphere is removed and the sediment is analyzed. There were conducted three experiences. The first is capillary sedimentation. The
second experiment is anodic deposition, with a graphite plate serves as an anode. The
third experiment is with pole reversal, when the upper (positive) electrode serves as
the anode and the graphite plate as the cathode. Figure 2 posted three photos of
deposition traces in three experiments. 250 A. Smetannikov and D. Onosov In the first experiment, there were matrix minerals practically absent, the number of
nanoparticles is minimal (Fig. 2A). In the second experiment, the number of matrix
minerals and nanoparticles is 1–2 orders of magnitude higher than in the first experi-
ment (Fig. 2B). In the third experiment, an intermediate result (Fig. 2C). The maxi-
mum of nanoparticles is fixed in the second experiment. Here, the deposition process is
enhanced by heating the graphite plate. Experiments have shown the natural properties of water in a capillary space change
created by a constant current source under the influence of an electric field. Capillary
solution acquires the properties of an electrolyte. The salt concentration in the sus-
pension is 0.5–1%. Fig. 2. A - sediment after capillary leaching; B - sediment after electrolytic anodic leaching;
C - sediment after cathodic leaching Fig. 2. A - sediment after capillary leaching; B - sediment after electrolytic anodic leaching;
C - sediment after cathodic leaching 3
Results and Discussion Extraction of micro- and nanoparticles from insoluble residues of salts, rocks, ores and
products of their processing is provided by creating an electric field. The field creates
the extracting properties of the electrolyte in suspension from the material of fine
fractions and salt solution. This factor ensures maximum extraction of micro- and nanoparticles and their
fixation on a graphite substrate for subsequent analysis is by the microprobe method. Moreover, it is possible to solve the direct problem - the extraction of micro- and
nanoparticles from insoluble residues of salts, ores and technogenic products, as well as
the inverse problem - “cleansing” any small fraction of the material from impurities of
minerals and metal particles. While using the capillary-electrolytic method of extracting micro- and nanoparti-
cles, the leaching effect (extraction) is achieved by 1–2 orders of magnitude higher than
the results of the application of the prototype. The main goal is to obtain information of
the form of finding elements-microimpurities. This result has been achieved. Resear-
ches are confirmed by the patent (Smetannikov and Onosov 2018). References Chueva MN (1950) Mineralogical analysis of schlich and ore concentrates. Gosgeolizdat,
Moscow, p 179 Deryagin BV, Churaev NV, Ovcharenko FD et al (1989) Water in disperse systems. Chemistry,
288 p Mitrofanov SI et al (1974) Mineral studies for enrichment. Nedra, Moscow, p 352 Oleynikova GA et al (2012) A nanotechnological method for extracting rhenium from rocks and
ores of black shale formations. Patent No. 245237 RU, St. Petersburg State University (RU),
Claims 12 June 2010. Publ. 10 July 2012, Bul. No. 19 Smetannikov AF (2014) Capillary method of extracting micro- and nanoparticles of minerals
from fine fractions for subsequent microprobe analysis. Yushkinsky readings 2014, Materials
of the mineralogical seminar with international participation, Syktyvkar, pp 177, 178 Smetannikov AF, Filippov VN (2010) Some features of the mineral composition of salt rocks and
products of their processing (for example, Verkhnekamsk salt deposit). Scientific readings of
the memory of P.N. Chirvinsky: Sat. articles, Perm, Issue 13, pp 99–113 Smetannikov AF, Onosov DV (2018) Capillary electrolytic method of extracting micro- and
nanoparticles of metal compounds from fine fractions of rocks, ores and industrial products. Patent №2659871, PFIC UB RAS (RU), Declared. 20 December 2016, Published 04 July
2018 Zhabreev VS (1997) Installation for the extraction of substances and particles from suspensions
and solutions. Patent 32098193 (RU), Chelyabinsk State Technical University, Appl. 26 July
1995, Publ. 10 December 1997 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. 4
Conclusions 1. The method relates to the methods for extracting micro- and nanoparticles of metal
compounds from various media under the influence of direct electric current in a
suspension of the material under study, water and salt, placed in a graphite substrate. Methods of Extraction of Micro- and Nanoparticles of Metal Compounds 251 2. The new method is different from the prototype by the use of an external electric
field, the intensity of extraction of nanoparticles and the completeness of extraction. 2. The new method is different from the prototype by the use of an external electric
field, the intensity of extraction of nanoparticles and the completeness of extraction. References If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Efficiency Evaluation for Titanium
Dioxide-Based Advanced Materials
in Water Treatment M. Harja1, O. Kotova2, S. Sun3, A. Ponaryadov2(&),
and T. Shchemelinina4 1 Gheorghe Asachi Technical University of Iasi, Iași, Romania
2 Institute of Geology Komi SC UB RAS, Syktyvkar, Russia
avponaryadov@geo.komisc.ru
3 Institute of Non-metallic Minerals, Department of Geological Engineering,
School of Environment and Resource, Southwest University of Science
and Technology, Mianyang, People’s Republic of China
4 Institute of Biology Komi SC UB RAS, Syktyvkar, Russia Abstract. We present a comparative evaluation of efficiency of titanium dioxide
polymorphs as an active photocatalyst (commercially available DegussaP25,
anatase (Sigma Aldrich), natural leucoxene concentrate (Pizhemskoe deposit,
Russia) and titanium dioxide nanotubes based on it). The materials obtained on
the basis of relatively inexpensive and affordable ilmenite-leucoxene ore have the
same efficiency as more expensive commercial products. Keywords: Anatase Ilmenite-leucoxene ores Nanotube Water treatment © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 255–258, 2019.
https://doi.org/10.1007/978-3-030-22974-0_61 2
Methods and Approaches Titanium dioxide. Degussa P25 (80% anatase, 20% rutile; Sigma Aldrich, France) was
used as photocatalyst without any purification. It has a BET surface (average) of 50 m2/
g, a particle size of 20–50 nm. Anatase (Sigma Aldrich) was used as photocatalyst
without any purification. It has a BET surface (average) of 80 m2/g, elongated particles
with a size of 15–30 nm. Leucoxene concentrate (LC) was obtained from the Pizhemskoe deposit (Russia). Chemical composition (wt%): TiO2 – 42.12, SiO2 – 46.57, Fe2O3 - 1.04, Al2O3 – 7.57,
K2O - 1.61, MnO - 0.06, CaO - 0.13, MgO - 0.37, SO3 - 0.06, P2O5 - 0.17, ZrO2 - 0.05,
NbO - 0.11. The particle size is about 20–40 mcm. Titanium dioxide nanotubes (TiNT) were obtained using a hydrothermal treat ment
procedure. The detailed description is given elsewhere (Kotova et al. 2016a). The photocatalytic activity of the samples was studied using a test reaction of
decomposition of trichlorophenol in Hereaus circular reactor of a volume of 350 cm3. Vertically to the reactor axis the TQ150 Z2 mercury lamp (150 W, 352–540 nm) was
located. The control solutions were analyzed by liquid chromatography (Hypersil C18
reverse phase HPLC column). The solvent was a solution of acetonnitrile in water in a
ratio of 3:2. The solvent flow was 0.5 ml/min. 1
Introduction In recent years, advanced oxidation processes have been proposed as alternative
methods for eliminating toxic organic pollutants from aquatic systems. Semiconductor
heterogeneous photocatalysis is one of the most promising and effective method. This
method is environment friendly, because the reaction products of the oxidation of
organic pollutants are carbon dioxide and water. A comparative analysis of the eco-
nomic efficiency of water purification showed that the photocatalytic method was the
cheapest (Duduman et al. 2018, Kotova et al. 2016b). The treatment of water from phenols, in particular, containing chlorine (2, 4, 6 -
trichlorophenol, TCP), is an important public health task because of their estrogenic,
mutagenic or carcinogenic effects. Their toxicity depends on the degree of chlorination
and the position of chlorine atoms in relation to the hydroxyl group. Removing these
compounds from the water is necessary to protect both human health and the envi-
ronment. To produce semiconductor photocatalyst based on titanium dioxide, multi-
stage synthesis methods are most often used, using orthotitanium acid or titanium
tetrachloride as precursors. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 255–258, 2019. https://doi.org/10.1007/978-3-030-22974-0_61 256 M. Harja et al. The aim of the work is the comparative evaluation of efficiency of commercially
available titanium dioxide (Degussa P25, Anatase Sigma Aldrich), natural (leucoxene
concentrate, Pizhemskoe deposit, Russia) and titanium dioxide nanotubes based on
natural leucoxene as active photocatalysts (TiNT). 3
Results and Discussion The initial leucoxen (Fig. 1A) is a mixture of two phases: rutile and quartz. The peaks
are clear, which indicates a high crystallinity of these phases. There are weak reflexes
of ilmenite and anatase. Leucoxen is a rutile microcrystalline matrix, saturated with the
finest inclusions of quartz (Ponaryadov 2017). The synthesized sample (Fig. 1B) is a
mixture of two phases: quartz and sodium titanate Na2Ti6O13. The chemical compo-
sition (semi-quantitative): TiO2 – 74.68%, SiO2 – 12.64%, Fe2O3 – 5.44%, Al2O3 –
4.71%, K2O – 0.93%. The structural rearrangement at the nanoscale level – formation of titanium di-oxide
nanotubes – leads to decreasing band gap: anatase – 3.1, LC –2.8, TiNT – 2.4 eV. Another important parameter is the specific surface area. During formation of titanium
dioxide nanotubes we observed increasing specific surface, which is associated with
formation of external and internal surfaces. For the studied samples, the specific surface
area was: anatase – 80, LC – 13, TiNT – 230 g/m2. Efficiency Evaluation for Titanium Dioxide-Based Advanced Materials 257 Fig. 1. XRD patterns for leucoxene concentrate, heavy fraction (A) and as-synthesized TiNT (B) Fig. 1. XRD patterns for leucoxene concentrate, heavy fraction (A) and as-synthesized TiNT (B) Kinetics of heterogeneous photooxidation reaction in liquid medium in the presence
of a catalyst is described by the Langmuir-Hinshelwood model. For the reaction of
decomposition of trichlorophenol, the time dependence ln(C0/C) is linear, at that the
slope ratio gives a constant kapp. The time dependence graphs for the studied samples
are presented in Fig. 2. Fig. 2. Curves of decomposition of trichlorophenol in water medium (
- leucoxene concentrate,
- anatase,
- DegussaP25, - titanium dioxide nanotubes) Fig. 2. Curves of decomposition of trichlorophenol in water medium (
- leucoxene concentrate,
- anatase,
- DegussaP25, - titanium dioxide nanotubes) The adsorption and decomposition reaction on surface occur simultaneously, most
likely, they do not determine the reaction rate. In the initial period of time (0–10 min),
trichlorophenol is adsorbed on the sample surface and the reaction rate increases. Upon
reaching the full coverage of the surface with adsorbate, the reaction rate is maximal
and does not change in the future. Based on the received data, the values kapp of
reaction constants were calculated: 0.005 for leucoxene concentrate, 0.006 for anatase,
0.025 for Degussa P25, 0.036 for titanium dioxide nanotubes. 3
Results and Discussion Thus, TiNT, produced by the hydrothermal method from ilmenite-leucoxene ore,
are competitive photocatalysts in water treatment from organically contaminants in
comparison to the above stated synthetic analogues. 258 M. Harja et al. 4
Conclusions We studied the dependence of the kinetics of photoinduced decomposition of
trichlorophenol in water solutions in the presence of various types of catalysts based on
titanium dioxide: commercially available DegussaP25 and anatase (Aldrich), leucoxene
concentrate (Pizhemskoe deposit), titanium dioxide nanotubes. We calculated reaction
constants of the photoinduced decomposition of trichlorophenol. It is shown that
advanced materials on the basis of relatively inexpensive and affordable ilmenite-
leucoxene ore have the same efficiency as expensive commercial products. Acknowledgements. This research was supported by UB RAS project №15-18-5-44 and project
AAAA-A17-117121270037-4 “Scientific basis for effective development and use of the mineral
resource base, development and implementation of innovative technologies, geological and
economic zoning of the Timan-North Ural region”. References Duduman CN, de Salazar y Caso de Los Cobos JMG, Harja M, Barrena Pérez MI, Gómez de
Castro C, Lutic D, Kotova O, Cretescu I (2018) Preparation and characterisation of
nanocomposite materials based on TiO2-Ag for environmental applications. Environ Eng
Manag J 17(4):2813–2821 Kotova O, Ozhogina E, Ponaryadov A, Golubeva I (2016b) Titanium minerals for new materials. In: IOP conference series: materials science and engineering, p 012025. https://doi.org/10. 1088/1757-899x/123/1/012025 Kotova OB, Ponaryadov AV, Gömze LA (2016a) Hydrothermal synthesis of TiO2 nanotubes
from concentrate of titanium ore Pizhemskoe deposit (Russia). Vestnik IG Komi SC UB RAS
1:34–36 Ponaryadov AV (2017) Mineralogical and technological features of ilmenite-leucoxene ores of
Pizhemskoe deposit, Middle Timan. Vestn Inst Geol Komi SC UB RAS 1:29–36. https://doi. org/10.19110/2221-1381-2017-1-29-36 (in Russian) Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 259–262, 2019.
https://doi.org/10.1007/978-3-030-22974-0_62 The Use of Karelia’s High-Mg Rocks
for the Production of Building Materials,
Ceramics and Other Materials
with Improved Properties V. Ilyina(&)
Institute of Geology KarRC RAS, Petrozavodsk, Russia
Ilyina@igkrc.ru V. Ilyina(&) Institute of Geology KarRC RAS, Petrozavodsk, Russia
Ilyina@igkrc.ru Abstract. The possible use of high-Mg host rocks, such as serpentinite and
pyroxenite, from the Aganozero chromium ore deposit and serpentinite from the
Ozerki soapstone deposit, Republic of Karelia, for the production of heat-
insulating building materials, ceramic pigments and filters for purification of
technogenous solutions is assessed. The results of analysis of the mineralogical
compositions of serpentinites and pyroxenite, as well as the physico-mechanical
properties (strength, heat conductivity, shrinkage upon roasting, moisture
resistance, etc.) and structural characteristics of the ceramic and heat-insulating
materials produced on their basis are reported. Keywords: Serpentinite Pyroxenite Ceramics Heat conductivity
Mechanical strength Facing material Keywords: Serpentinite Pyroxenite Ceramics Heat conductivity
Mechanical strength Facing material Keywords: Serpentinite Pyroxenite Ceramics Heat conductivity
Mechanical strength Facing material 2
Methods and Approaches The mineral composition of the analyzed samples was studied in the IG KarRC RAS by
optical microscopy methods, X-ray phase analysis (XPA) and thermal analysis (TA). Rock-forming minerals were studied by Vega II LSH scanning electron microscope
with INCA Energy 350 energy dispersion analyzer. X-ray phase analysis was per-
formed by ARL X’TRA diffractometer with CuKl radiation. The physico-mechanical
properties of the materials and ceramics were assessed in accordance with State All-
Russia standards. 1
Introduction Non-conventional high-Mg rocks can be used for the production of ceramic, building
and other materials because they are widespread but mainly because of the chemical,
mineral and structural characteristics of their mineral constituents: periclase, forsterite,
diopside, augite, enstatite and serpentine. The main high-Mg mineral constituents of
ceramics suffer phase transformations upon heating, as a result of the disintegration and
recrystallization of their lattice (Deere et al. 1965), forming crystalline phases that
improve the physic-mechanical properties of ceramics. As forsteritic ceramics suffers
no polymorphic transformations, it does not age and is mechanically strong. It is used
for the production of dielectrics, heat-insulating materials, facing ceramics and filters
for water purification. The aim of the present project is to study serpentinites, kemistites
(rocks of hydrotalcite-serpentine composition) and pyroxenites that host chromium ores
at the Aganozero deposit and serpentinites at the Ozerki soapstone deposit and to use
them for the production of ceramics with a dominant forsteritic crystalline phase, filters
for purification of technogenic solutions and ceramic pigments. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 259–262, 2019. https://doi.org/10.1007/978-3-030-22974-0_62 V. Ilyina V. Ilyina 260 Strength
Heat conducƟvity coefficient Fig. 1. Dependence of variations in the properties of ceramics on kemistite concentration Fig. 1. Dependence of variations in the properties of ceramics on kemistite concentration 4
Conclusions The materials developed display a porous structure and, consequently, low density, low
heat conductivity and high strength. Hence, they can be widely used in industrial and
civil engineering for the insulation of buildings, for the heat insulation of the hot
surfaces of equipment (furnaces and pipelines), for intensifying high temperature
processes and for fuel saving. Ilyina VP, Shchiptsov VV, Frolov PV (2013) Raw mixture for the pro-duction of a porous heat-
insulating material, Bull no 31 Patent no 2497774 RF, MPC SO4B 33/132 Deere WA, Haui RA, Zoosman J (1965) Rock-forming minerals. Chain silicated. vol 2, p 405
Il’ina VP, Inina IS, Frolov PV (2017) Ceramic mix based on pyroxenite and low-melting clay.
Glass and Ceramics pp 1–4 3
Results and Discussion Differences in the mineral composition of serpentinites affect their chemical compo-
sition. Aganozero kemistites and serpentinites are the richest in magnesium (36–38%)
and contain minor quantities of impurities (0.1–0.5% Al2O3 and 0.24–0.5% CaO) and
elevated quantities of crystallization water (loss on ignition is 15–18.5%). Ozerki
serpentinite is rich in MgO (36.92%) and contains Al2O3 (2.2) and CaO (0.22%) as
impurities. All the samples are iron-rich (3.46–10.02% Fe2O3, 1.72–3.9% FeO). The
mineralogo-analytical study (RPA, TA) of serpentinites showed that Ozerki serpen-
tinites consisted of 89.1% fine-grained lamellar antigorite aggregate and that Agano-
zero serpentinites contained 78% lizardite. Ore minerals are represented by magnetite
and ilmenite. The main minerals of natural pyroxenite (wt%) are augite (67, 2), for-
sterite (4, 3), enstatite (23, 7) and serpentine (4, 8). Kemistite-based porous heatinsu-
lating ceramics was developed and its properties were studied (Patent no. 2497774,
2013). Electron microscopy study has shown that forsterite, produced by serpentine
recrystallization, is the main crystal-line phase of heat-insulating ceramics. The prop-
erties of heat-insulating ceramics are shown in Fig. 1. Pyroxenite (20–70%) and hydromica-based facing ceramics (Il’ina et al. 2017)
displays water absorption of 13–15.8% at a roasting temperature of 900–1100 °C,
which is consistent with the current standards. At 1200 °C, the water absorption of all
the masses decreases rapidly from 0 to 1.2%. Their bending strength is 9–10 MPa. Pyroxenite-based ceramic pigment has been developed. Unlike the well-known
pigment, it can be used to obtain a stable color after roasting over a wide temperature of
750–1250 °C. The results obtained (Ilyina et al. 2018) show that Ozerki serpentinite can be used
for the production of an Mg-silicate reagent for the removal of heavy metals from
solutions, e.g. the removal of heavy metal compounds from highly polluted techno-
genous solutions by filtration through loading from a granulated reagent. The Use of Karelia’s High-Mg Rocks for the Production of Building Materials 261 0.02
0.025
0.03
0.035
0.04
0
5
10
15
20
25
30
58
60
62
64
66
68
70
72
Heat conducƟvity coefficient,
Wt/mk
Bending strength, Mpa
KemisƟte concentraƟon, mas.%
Strength
Heat conducƟvity coefficient
Fig. 1. Dependence of variations in the properties of ceramics on kemistite concentration 0.02
0.025
0.03
0.035
0.04
0
5
10
15
20
25
30
58
60
62
64
66
68
70
72
Heat conducƟvity coefficient,
Wt/mk
Bending strength, Mpa
KemisƟte concentraƟon, mas.% References Deere WA, Haui RA, Zoosman J (1965) Rock-forming minerals. Chain silicated. vol 2, p 405
Il’ina VP, Inina IS, Frolov PV (2017) Ceramic mix based on pyroxenite and low-melting clay. Glass and Ceramics pp 1–4 Deere WA, Haui RA, Zoosman J (1965) Rock-forming minerals. Chain silicated. vol 2, p 405
Il’ina VP, Inina IS, Frolov PV (2017) Ceramic mix based on pyroxenite and low-melting clay. Glass and Ceramics pp 1–4 Ilyina VP, Kremenetskaya IP, Gurevich BI, Klimovskaya EE, Ivashevskaya SN (2018) The study
of serpentinized ultramafics from the Kareli-an-Kola Region and the production of a Mg-rich-
silicate reagent on their basis for the removal of heavy metals from solutions. In: 18th
International Multidisciplinary Scientific GeoConference SGEM2018: Conference proceed-
ings. STEF 92 Technology Ltd., 51 “Alexander Malinov” Blvd., 1712 Sofia, Bulgaria,
Energy and Clean Technologies Issue: 4, 2. 2 July – 8 July 2018. Albena, Bulgaria, vol 18
(13), pp 207–213 Ilyina VP, Shchiptsov VV, Frolov PV (2013) Raw mixture for the pro-duction of a porous heat-
insulating material, Bull no 31 Patent no 2497774 RF, MPC SO4B 33/132 V. Ilyina 262 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Kinetic Features of Formation
of Supramolecular Matrices on the Basis
of Silica Monodisperse Spherical Particles D. Kamashev(&)
Institute of Geology Komi SC UB RAS, Syktyvkar, Russia
kamashev@geo.komisc.ru D. Kamashev(&) Institute of Geology Komi SC UB RAS, Syktyvkar, Russia
kamashev@geo.komisc.ru Abstract. We have established that under such conditions, when the formation
of a supramolecular structure from spherical silica particles 220–320 nm in
diameter is limited by the rate of introduction of particles into the sedimentation
zone (sedimentation deposition in a constant cross section tube), the particle
deposition rate, as well as the formation rate of the supramolecular structure, is
strictly linear. At the same time, under conditions of excess of disperse phase in
the zone of formation of the supramolecular structure (sedimentation deposition
in a tube with a modified cross section), the deposition rate is also linear, but
there is some delay in formation of the supramolecular structure in time, the
larger the smaller is the particle size of the disperse phase. After the formation of
the supramolecular structure is completed, a region with an increased concen-
tration of the disperse phase remains, the height of which is greater, the smaller
is the particle size. It is shown that a certain concentration of the disperse phase
is necessary to begin the formation of a supramolecular structure, below which
the formation of a supramolecularly ordered structure does not occur. The
concentration of the disperse phase, necessary for beginning of formation of a
supramolecular structure, is a constant value that does not vary with time and
depends only on the size of the particles. Keywords: Supramolecular structure Monodisperse spherical silica particles Keywords: Supramolecular structure Monodisperse spherical silica particles © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 263–266, 2019.
https://doi.org/10.1007/978-3-030-22974-0_63 1
Introduction The supramolecularly ordered structures, based on monodisperse spherical silica par-
ticles, generated interest relatively long ago, back in the 70s of the last century, in
connection with attempts to synthesize artificial analogs of noble opal on their basis
(Stober et al. 1968). The overwhelming majority of studies of that time were aimed at
developing conditions for synthesis of spherical silica particles and supramolecular
structures based on them, and ended with the development of methodological bases for
production of synthetic noble opals (Deniskina et al. 1987). However at present the
supramolecular silica particles are more often considered as promising objects for
synthesis of new composition materials, photonic crystals and nanostructured materials
based on them. The 3D ordered closest packing of monodisperse silica spheres is an
ideal matrix for creating a wide class of new nanostructured materials, but it greatly
increases requirements for the monodispersity of particles, their size and the defects of D. Kamashev 264 derived supramolecular structures (Kamashev 2012). With this aim and within the task
to develop the basis of synthesis of supramolecularly ordered matrices we carried out
experimental studies of the rate of precipitation of silica particles and formation of a
supramolecular structure on their basis in various conditions (growth modes). 2
Methods and Approaches Monodisperse spherical silica particles with radius 109, 138 and 158 nm were syn-
thesized by Stober-Fink method (Stober et al. 1968). The sizes of the obtained particles
were determined by Photocor Complex dynamic light scattering spectrometer at a laser
wavelength 661 nm, a scattering angle 90°, and a correlation function accumulation
time 20 min. The initial concentration of the dispersed phase particles was about
2 wt.%. The deposition rate of the silica particles was measured in glass tubes 750 mm
high and 20 mm in diameter, with the control of deposition boundary advancing every
30 days with accuracy 0.5 mm. The average daily temperature was also taken into
account to calculate the temperature correction associated with the expansion of the
dispersion environment. The following values were obtained for the silica particles of
different radius (Fig. 1). Fig. 1. Dependence of height of deposition of silica particles of different sizes over time. The
coefficient before x (0.201805, 0.168749 and 0.149001) in the equation of approximating curve
represents the rate of particle deposition, cm/day. Fig. 1. Dependence of height of deposition of silica particles of different sizes over time. The
coefficient before x (0.201805, 0.168749 and 0.149001) in the equation of approximating curve
represents the rate of particle deposition, cm/day. To measure the rate of formation of the supramolecular structure, the deposition of
particles was carried out in glass tubes with a narrowing in the lower part, which
reduced the area (concentration of suspension) by about 80 times. This mode of for-
mation of the supramolecular structure is characterized by a constant increased content
of the dispersed phase, unlike deposition in tubes with a constant cross section, where
the formation of region with a high content of particles is limited by their deposition
rate. The obtained data on the rate of formation of a supramolecularly ordered structure
are presented in Fig. 2. At such rates one monolayer of silica particles with radius
158 nm is formed approximately within 90 s. Kinetic Features of Formation of Supramolecular Matrices 265 Fig. 2. Dependence of height of a supramolecular structure formed from silica particles of
different size over time. The coefficient before “x” (0.28115, 0.22990 and 0.20110) in the
equation of approximating curve is a growth rate of the supramolecular structure in conditions of
high concentration (excess) of the dispersed phase, mm/day. Fig. 2. 2
Methods and Approaches Dependence of height of a supramolecular structure formed from silica particles of
different size over time. The coefficient before “x” (0.28115, 0.22990 and 0.20110) in the
equation of approximating curve is a growth rate of the supramolecular structure in conditions of
high concentration (excess) of the dispersed phase, mm/day. 4
Conclusions The obtained experimental data on deposition rate of monodisperse spherical silica
particles with formation of a supramolecular structure showed that its formation was
the result of a kind of second-order phase transition (self-organization of particles). This transition was associated with a threshold concentration of particles of dispersed
phase in the area of deposition, below which the formation of the supramolecular
structure did not occur. When this value was exceeded, the process of formation of a
supramolecular structure started, and the rate of its formation was strictly linear and
independent of the concentration of the dispersed phase. If for some reason the
threshold concentration of particles was not reached (for example, very small sizes, low
density of particles or high viscosity of the dispersion medium), then the formation of a
supramolecular structure did not occur. Acknowledgements. The work was accomplished with partial financial support by UB RAS
program No. 15-18-5-44, and RFBR No. 19-05-00460a. 3
Results and Discussion Our data testify to that the particle deposition rate (Fig. 1) is linear and starts at the
origin and strictly obeys the Stokes equation throughout the time. In turn, the analysis
of data on the formation rate of the supramolecular structure (Fig. 2) shows that the
formation rate of the ordered structure is also linear, however, the process of
supramolecular crystallization does not begin immediately, but after some time, due to
creation of “supersaturation” or some increased concentration of particles of dispersed
phase in the bottom region. This time depends on size of particles and increases with
their decrease. As a result of deposition of monodisperse silica particles and formation of a
supramolecular structure we noted that initially a zone with an increased concentration
of dispersed phase was formed in the bottom region. The boundaries of this zone were
clearly expressed, it possessed a constant height, both in the process of the beginning of
deposition of particles, and after its completion. The height of the condensed y = -0.0125x + 3.037
R² = 0.9909
0
2
4
6
8
10
0
50
100
150
200
Height of thickened
suspension, mm
Radius of the particles, nm
Fig. 3. Dependence of thickened suspension height on the particle size of the disperse phase
after completion of deposition. Fig. 3. Dependence of thickened suspension height on the particle size of the disperse phase
after completion of deposition. D. Kamashev 266 suspension zone after deposition of particles was constant and depended only on size of
dispersed particles (Fig. 3) with the exception of particles of radius less than 60 nm. In
this case a similar zone with clearly defined boundaries was not formed, and the
supramolecular structure was not formed. Three-Cation Scandium Borates
RxLa1−XSc3(BO3)4(R = Sm, Tb):
Synthesis, Structure, Crystal Growth
and Luminescent Properties A. Kokh1(&), A. Kuznetsov1, K. Kokh1,2, N. Kononova1,
V. Shevchenko1, B. Uralbekov3, A. Bolatov3, and V. Svetlichnyi4 1 Sobolev Institute of Geology and Mineralogy SB RAS, Novosibirsk, Russia
a.e.kokh@gmail.com
2 Novosibirsk State University, Novosibirsk, Russia
3 Al-Farabi Kazakh National University, Almaty, Kazakhstan
4 Tomsk State University, Tomsk, Russia 1 Sobolev Institute of Geology and Mineralogy SB RAS, Novosibirsk, Russia
a.e.kokh@gmail.com
2 Novosibirsk State University, Novosibirsk, Russia
3 Al-Farabi Kazakh National University, Almaty, Kazakhstan
4 Tomsk State University, Tomsk, Russia Abstract. Complex ortohoborates of rare earth metals with the general
chemical formula RxLa1−xSc3(BO3)4 (R = Sm, Tb) have been obtained by solid
state synthesis and spontaneous crystallization. These crystals belong to the
huntite family with the space group R32 and for x = 0.5 have unit cell
parameters a = 9.823(6), c = 7.975(3) (SLSB) and a = 9.803(3), c = 7.960(4) Å
(TLSB). Keywords: Crystal Borate Structure Huntite Growth Luminescence References Deniskina ND, Kalinin DV, Kazantseva LK (1987) Noble opals. Novosibirsk, Nauka, Sib. otd. (Trudy institute geologii I geofiziki) 693, p 184 Kamashev DV (2012) Synthesis, features and model of formation of supramolecular silica
structures. Phys Chem Glass 38(3):69–80 Stober W, Fink A, Bohn E (1968) Controlled growth of monodisperse silica spheres in the
micron size range. J Colloid Interface Sci 26:62–69 Stober W, Fink A, Bohn E (1968) Controlled growth of monodisperse silica spheres in the
micron size range. J Colloid Interface Sci 26:62–69 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 267–271, 2019.
https://doi.org/10.1007/978-3-030-22974-0_64 1
Introduction Orthoborates with the general formula RX3(BO3)4, where R = Y, Ln; X = Al, Ga, Sc,
Cr, Fe are practically important and interesting from the point of view of crystal
chemistry objects for research. One of the important properties of these compounds is
the ability to form a non-centrosymmetric structure, which is called huntite-like. Such a
structure causes, for example, non-linear optical properties. To understand the formation of the huntite-like structure of three-cation scandobo-
rates, we consider the lanthanum – scandium borate LaSc3(BO3)4. The authors (He et al. 1999) distinguish three modifications of this crystal: high-temperature monoclinic with
the C2/c space group, medium temperature trigonal with the R32 space group (huntite-
like) and low-temperature monoclinic with the Cc space group. As a result of our
research (Fedorova et al. 2013) identity of the X-ray patterns of polymorphic modifi-
cations high and low was shown. The stabilization of the huntite-like structure can occur if an additional isomorphic
cation is introduced into the LaSc3(BO3)4 structure, that was confirmed in (Li et al. 2001) who initiated the new three-cation scandoborate with the huntite-like structure
NdxLa1−xSc3(BO3)4. Further, in a number of works by adding a third cation © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 267–271, 2019. https://doi.org/10.1007/978-3-030-22974-0_64 A. Kokh et al. 268 RxLayScz(BO3)4 nonlinear optical crystals with a stable huntite-like structure were
obtained with R = Gd (Xu et al., 2011); Y (Ye et al. 2005) and Lu (Li et al. 2007). The existence of a huntite-like structure for the boundary members of the REE series
suggests the stability of such a structure with the rest of the REE. This paper presents
data on the huntite-like structures SLSB and TLSB for systems RxLa1−xSc3(BO3)4
(R = Sm, Tb). 2
Methods and Approaches Polycrystalline sample of RxLa1−xSc3(BO3)4 (x = 0–0.5)were prepared by the method
of two stage solid state synthesis in a Pt crucible. The stoichiometric mixtures of pure
raw La2O3, Sc2O3, H3BO3 and R2O3(R = Sm, Tb) reactants were heated at 800 °C for
5 h to decompose H3BO3. At the second stage, the mixtures were grinded in an agate
mortar and heated again at 1300 °C for 12 h until the powder X-ray method showed no
peaks of initial compounds (Fig. 1). Fig. 1. X-ray pattern of SmxLa1−xSc3(BO3)4, where x = 0(a), 0.2(b), 0.3(c), 0.4(d), 0.5(e), 1(f). I- huntite (R32), X- monoclinic (C2/c) structure. Fig. 1. X-ray pattern of SmxLa1−xSc3(BO3)4, where x = 0(a), 0.2(b), 0.3(c), 0.4(d), 0.5(e), 1(f). I- huntite (R32), X- monoclinic (C2/c) structure. Spontaneous crystals of RxLa1−xSc3(BO3)4with dimensions 30 30 10 mm
with a transparent area of 5 5 5 mm were grown from LiBO2- LiF flux Fig. 2. A Pt crucible containing R0.5La0.5Sc3(BO3)4, Li2CO3, H3BO3 and LiF in the molar
ratio of 1:1,5:1,5:3 was heated to 1000 °C. The charge was held in a melted state for a
day to achieve homogenization. After this stage a platinum wire with a loop was placed
in the center of the melt surface and the temperature was decreased to 900 °C. Then the
melt was cooled with the 2 °C/day to 850 °C and following cooling at the rate of Three-Cation Scandium Borates RxLa1−XSc3(BO3)4(R = Sm, Tb) Three-Cation Scandium Borates RxLa1−XSc3(BO3)4(R = Sm, Tb) 269 15 °C/day to room temperature. The crystal was chosen for x-ray analysis. Powder
diffraction patterns were refined using the Rietveld method within the GSAS- II
program. Fig. 2. Crystals grown from LiBO2-LiF flux: TLSB (left) and SLSB (right) Fig. 2. Crystals grown from LiBO2-LiF flux: TLSB (left) and SLSB (right) The chemical composition of obtained crystals was measured by X-ray fluorescent
analysis using XRF 1800 (Shimadzu, Japan). The results of the analysis are conformed
with the formula obtained after crystal structure refinement: (Table 1) e 1. Composition of TLSB and SLSB based on X-ray fluorescent elemental analysis Table 1. Composition of TLSB and SLSB based on X-ray fluorescent elemental analysis
Composition of the TbxLayScz(BO3)4
Ratio of Tb/La
Starting melt Tb0.5La0.5Sc3(BO3)4
1
Center
Tb0.22La0.78Sc3(BO3)4
0.28
Edge
Tb0.24La0.75Sc2,99(BO3)4 0.32
Composition of the SmxLayScz(BO3)4
Ratio of Sm/La
Starting melt Sm0.5La0.5Sc3(BO3)4
1
Center
Sm0.32La0.69Sc2,98(BO3)4 0.46
Edge
Sm0.35La0.68Sc2,97(BO3)4 0.52 3
Results and Discussion Structure. According to Rietveld refinement both SLSB and TLSB crystalize in the
trigonal space group R32 with unit cell parameters: a = 9.823(6), c = 7.975(3) (SLSB)
and a = 9.803(3), c = 7.960(4)Å (TLSB). The structure framework is composed of the
R, La atoms, Sc atoms and B atoms occupy trigonal prisms, octahedra and planar
triangle of oxygen, respectively. The isolated (R, La)O6 trigonal prisms alternate along
the c-axis with BO3 triangle that are perpendicular to the c-axis. ScO6octahedra link to
each other along the edge and form twisted chain along c, which separate (R, La)O6
prisms as well. The discrepancies between refined diffraction spectra with model cal-
culations can be explained by crystal cleavage along {202} and {113}. A. Kokh et al. 270 Luminescence. Typical excitation and luminescence spectra of SLSB are shown
on Fig. 3(a). The strongest excitation peak of samarium crystal corresponds to
6H5=2 ! 4F7=2 transition located at 407 nm. Whereas luminescent spectrum of SLSB
has some peaks corresponding to4G5=2 ! 6HJ (J = 5=2, 7=2, 9=2 и 11=2) and located
at 566, 602, 645 and 708 nm. Fig. 3. Luminescent properties of (a) SLSB, (b) TLSB Fig. 3. Luminescent properties of (a) SLSB, (b) TLSB On Fig. 3(b) TLSB excitation and luminescence spectra are shown with a wide strip
at 300 nm corresponding to4F - 5D transition. Luminescent spectra consist of 5 peaks at
490, 505, 585 and 640 corresponding to5D4 - 7FJ (J = 6, 5, 4, 3) transitions. 4
Conclusions The formation of a huntite structure in systems RxLa1−xSc3(BO3)4, (R = Sm, Tb), as
well as the dependence of the compositions stable in the required structure depending
on the production method is shown. The spectral characteristics confirm the potential of
using crystals as luminescent materials. Acknowledgements. This work is supported by RFBR project#19-05-00198a, state contract of
IGM SB RAS and partially by Project GF MES RK IRN AP05130794. Xu X, Ye N (2011) GdxLa1−xSc3(BO3)4: a new nonlinear optical crystal. J Cryst Growth
324:304–308
Ye N, Stone-Sundberg JL, Hruschka MA et al (2005) Nonlinear Optical Crystal YxLayScz(BO3)4
(x + y + z = 4). Chem Mater 17:2687–2692 Three-Cation Scandium Borates RxLa1−XSc3(BO3)4(R = Sm, Tb)
271 References Fedorova MV, Kononova NG, Kokh AE, Shevchenko VS (2013) Growth of MBO3 (M = La, Y,
Sc) and LaSc3(BO3)4 Crystals from LiBO2–LiF Fluxes. Inorg Mater 49:482–486 )
3(
3)
y
g
He M, Wang G, Lin Z et al (1999) Structure of medium temperature phase b-LaSc3(BO3)4
crystal. Mater Res Innov 2(6):345–348 y
Li W, Huang L, Zhang G, Ye N (2007) Growth and characterization of nonlinear optical crystal
Lu0.66La0.95Sc2.39(BO3)4. J Cryst Growth 307:405–409 Li Y, Aka G, Kahn-Harari A, Vivien D (2001) Phase transition, growth, and optical properties of
NdxLa1−xSc3(BO3)4 crystals. J Mater Res 16:38–44 271 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Rational Usage of Amorphous Varieties
of Silicon Dioxide in Dry Mixtures of Glass
with Specific Light Transmittance N. Min’ko(&) and O. Dobrinskaya Belgorod State Technological University named after V G Shukhov,
Belgorod, Russia
minjko_n_i@mail.ru Abstract. The paper studied high-silicon amorphous rocks from the perspec-
tive of their application for glass production of different purpose. The results
contained data of calculation of dry mixtures for producing heat-protective glass
using amorphous varieties of silicon dioxide. The obtained glass specimens were
melted and studied for spectral characteristics. Keywords: Amorphous silicon dioxide Light transmittance Dry mixture
Heat-protection glass 1
Introduction High-silicon amorphous rocks as raw materials have a wide range of valuable features. Primarily, this is the amorphous (metastable) state of silicon dioxide (Kondrashov and
Kondrashov 2013). Moreover, one can distinguish some peculiarities of amorphous
varieties that can be regarded as drawbacks (Manevich et al. 2012): – instability of chemical composition; – SiO2 is accompanied by other components (up to 40%) that can play the role of
auxiliary raw materials; – SiO2 is accompanied by other components (up to 40%) that can play the role of
auxiliary raw materials; – increased content of aluminum oxides and iron. One of the main glass spectral characteristics is light transmittance. The main
components that affect light transmittance of glass products and that should be strictly
controlled are oxides of coloring metals that are encountered in conventional raw
materials (dolomite, feldspar concentrate, sands). These compounds include iron oxi-
des; their content in glass is strictly regulated: – sheet glass – 0.09–0.12%; – heat-protective – 0.6–0.7%; – clear container glass – 0.1 ± 0.01%; – brown container glass – 0.8 ± 0.1%; – green container glass – not regulated. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 272–276, 2019. https://doi.org/10.1007/978-3-030-22974-0_65 © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 272–276, 2019. https://doi.org/10.1007/978-3-030-22974-0_65 273 Rational Usage of Amorphous Varieties of Silicon Dioxide The composition of amorphous varieties of silicon dioxide (Distantov 1976) is
characterized by increased content of iron oxides (Table 1), which impedes their wide
application in production of glass articles. The composition of amorphous varieties of silicon dioxide (Distantov 1976) is
characterized by increased content of iron oxides (Table 1), which impedes their wide
application in production of glass articles. Table 1. Variation of chemical composition of amorphous silicic raw materials Table 1. 1
Introduction Variation of chemical composition of amorphous silicic raw materials
Rock
Content of oxides [wt.%]
SiO2
Al2O3
Fe2O3
CaO
MgO
K2O
Na2O
TiO2
Diatomites 73.0–90.0
3.3–7.5
2.0–5.2
less than
0.6
0.6–1.7
less than
1.0
less than
0.5
less than
0.3
Opokas
52.1–91.4
2.5–15.4
1.0–5.0
0.43–17.1
less than
2.48
0.6–4.0
0.1–1.0
less than
0.2
Pearlites
68.5–75.3
11.2–16.3
less than
3.0
0.5–2.0
less than
1.0
1.5–4.0
2.0–6.2
0.1–0.5
Tripolites
35.3–86.7
2.5–11.6
0.3–3.4
0.4–31.2
0.2–1.6
0.85–2.1
less than
0.5
less than
0.2 Taking into account increased content of iron and aluminum oxides, these rocks
can be used as aluminum- or iron-containing raw materials in production of heat-
protection glass or dark-glass containers. 2
Methods and Approaches Current work assesses amorphous silicon dioxide (ASD) as aluminum-containing raw
material that can partially replace quartz sand and other conventional raw materials
(Table 2). Table 2. Composition of average samples of amorphous silicon dioxide varieties Table 2. Composition of average samples of amorphous silicon dioxide varieties
Rock
Content of oxides [wt.%]
SiO2
Al2O3 Fe2O3 CaO
MgO
K2O Na2O TiO2
Diatomite 87.22
6.79
2.22
0.431 1.25
1.08 0.245 0.265
Opoka
92.47
3.38
1.32
0.506 0.712 0.88 –
0.174
Pearlite
72.0
16.45
1.06
0.863 0.422 4.30 4.21
0.145
Tripolite
79.93 10.99
3.17
0.838 1.81
1.99 0.273 0.687 To conduct experimental studies, the dry mixtures for heat protective glass with
ASD were calculated. The glass was melted from the dry mixtures using pearlite. The
content of iron oxides in pearlite is insufficient for production of heat-protective glass,
which necessitates the introduction of iron containing material such as magnetite. The glass was melted in an electric kiln with silicon carbide heating elements at
maximum melting temperature of 1420 °C. Another batch of glass was melted without
pearlite. Spectral light transmittance in the visible range was measured automatically on SF-
56 spectrophotometer (Russia). The specimens were prepared by mechanical grinding
and polishing on laboratory setups. 274 N. Min’ko and O. Dobrinskaya N. Min’ko and O. Dobrinskaya 3
Results The results showed that the application of ASD for preparation of dry mixtures allowed
reducing number of conventional materials for glass melting (Table 3). Table 3. Economy of conventional raw materials after replacement by ASD, %
Rock
Raw material
Sand Soda
Creta Dolomite
Opoka
72.7
2.44
1.61 19.71
Pearlite
43.8
20.15 38.7
40.38
Diatomite 41.7
6.5
4.08 25.14
Tripolite
27.6
7.79
7.66 27.06 Table 3. Economy of conventional raw materials after replacement by ASD, % The glass produced in the laboratory differed in color (Fig. 1), which depends on
the shift of equilibrium Fe2+$Fe3+. 1
2
3
Fig. 1. Specimens of sheet glass: 1 – with pearlite; 2 – with pearlite and magnetite; 3 – with
pearlite and magnetite (+ coal) Fig. 1. Specimens of sheet glass: 1 – with pearlite; 2 – with pearlite and magnetite; 3 – with
pearlite and magnetite (+ coal) We studied spectral characteristics of the glass specimens: sheet glass melted from
conventional components; sheet glass with addition of pearlite as aluminum-containing
raw material; sheet glass with addition of magnetite (magnetite was reduced by coal)
(Fig. 2). T
λ [nm]
1
2
3
4
Fig. 2. Spectral light transmittance of sheet glass: 1 – conventional raw materials; 2 – with
pearlite; 3 – with pearlite and magnetite; 4 – with pearlite and magnetite (+ coal) Fig. 2. Spectral light transmittance of sheet glass: 1 – conventional raw materials; 2 – with
pearlite; 3 – with pearlite and magnetite; 4 – with pearlite and magnetite (+ coal) 275 Rational Usage of Amorphous Varieties of Silicon Dioxide The data from Table 4 demonstrated that in the visible spectrum, the light trans-
mittance of the glass with application of pearlite differed from glass melted with
conventional raw materials. The highest light transmittance was shown by sheet glass
melted from conventional raw materials, since the content of iron in quartz sand was
minimal (0.022 wt%). Table 4. Light transmittance of sheet glass specimens calculated for the thickness of 10 mm
Specimen
Light transmittance T [%]
Diathermancy
index = 10−1
T1100
k = 570 nm
k = 1100 nm
As per
equation
As per
nomogram*
As per
equation
As per
nomogram*
1
88.6
88.1
67.4
67.7
6.7
2
48.4
48.8
8.4
8.2
0.8
3
30.8
30.5
5.1
4.8
0.4
4
12.3
12.0
3.7
3.2
0.3
* - Amosov’s nomogram Table 4. 3
Results Light transmittance of sheet glass specimens calculated for the The
heat-protective
characteristics
are
assessed
by
light
transmittance
at
k = 1100 nm. In the infrared range of the spectrum, the light transmittance of speci-
mens 2, 3 and 4 reduces due to the presence of impurities of Fe2+, which provisions the
heat-protective characteristics. In IR-range, the light transmittance of studied glasses is different (from 3 to 67%),
i.e. high heat-protective properties are possessed by specimens 3 and 4, the IR-light
transmittance is 3–5%. However, the production of glass with such content of FeO is
unreasonable due to low light transmittance in the visible spectrum. Such dry mixtures
can be used for special glass. 4
Conclusions The calculations allowed determining the amorphous varieties of silicon dioxide that
could replace most conventional materials in dry mixtures. They were: opoka (as a
replacement for quartz sand) for both aluminum- and iron-containing raw materials,
and pearlite capable of replacing the biggest amount of soda, creta and dolomite when
using it as an iron-containing material. Additional studies of glass properties produced from dry mixtures with pearlite
were required (spectral characteristics, crystallization capacity and impact of redox
conditions of melting). Nevertheless, the work showed that pearlite could be used in the
technology of heat-protective glass. 276 N. Min’ko and O. Dobrinskaya Acknowledgments. The work is realized in the framework of the Program of flagship university
development on the base of Belgorod State Technological University named after V.G. Shukhov,
using the equipment of High Technology Center at BSTU named after V.G. Shukhov. Distantov AG (1976) Silicon rocks of USSR. Tatar Press, Kazan
Kondrashov VI, Kondrashov DV (2013) Perspectives of synthesis of industrial compositions of
float-glass on the basis of crystalline and amorphous silicon dioxide. GlassRussia. 3:31–33
Manevich VE, Subbotin RK, Nikiforov EA, Senik NA, Meshkov AV (2012) Diatomite as silica-
containing material for glass industry. Glass Ceram 5:34–39 References Distantov AG (1976) Silicon rocks of USSR. Tatar Press, Kazan
Kondrashov VI, Kondrashov DV (2013) Perspectives of synthesis of industrial compositions of
float-glass on the basis of crystalline and amorphous silicon dioxide. GlassRussia. 3:31–33
Manevich VE, Subbotin RK, Nikiforov EA, Senik NA, Meshkov AV (2012) Diatomite as silica-
containing material for glass industry. Glass Ceram 5:34–39 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Peculiarities of Phase Formation in Artificial
Ceramic Binders for White-Ware Compositions
I. Moreva(&), E. Evtushenko, O. Sysa, and V. Bedina
Belgorod State Technological University named after V G Shukhov,
Belgorod, Russia
moreva_bstu@mail.ru Belgorod State Technological University named after V G Shukhov,
Belgorod, Russia
moreva_bstu@mail.ru Abstract. The production of sanitary white wares traditionally uses multi-
component mixes, which is necessitated by a whole complex of properties: high
density and low humidity of molding slurries with low thixotropic strengthening
and good filterability of the slurry. However, modern understanding of the
structure and properties of materials open opportunities for optimization of
technological process and production of higher-quality articles. The imple-
mentation of activation technologies and replacement of traditional molding
slurries by artificial ceramic binders will reduce the number of components in
mixes and optimize the production of white wares. Since the achievement of
working performance of ceramic materials substantially depends on the phase
formation processes in a material during sintering. Current work analyses the
phase transformations occurring at all stages of white ware production as per
proposed technology. Keywords: Artificial ceramic binders Phase conversions Porcelain
Pottery Ceramic slurry Keywords: Artificial ceramic binders Phase conversions Porcelain
Pottery Ceramic slurry 2
Methods and Approaches Initial materials were broken, mixed and wetted to 15–20% with consequent formation
of bricks. After drying, the bricks were burnt at the temperature of 950–1000 °C with
shock cooling for increased system activity. The thermally activated mix was milled as
per the technology of highly concentrated ceramic binders for suspensions with intro-
duction of electrolytes and liquid phase at the loading of the first part of the material. The XRD analysis of the investigated materials was made on DRON-3 diffractometer. The diffractograms were recorded with filtered CuKa-radiation (Ni filter); the tube
voltage was 20 kV; tube anode current was 20 mA; the measurement limit was 10,000–
4,000 pulses per second; the detector rotation speed was 2.4 min−1; the angular mark
was 10. The phases were identified using JCPDF cards. The samples with the dimen-
sions of 30 30 30 mm from obtained suspensions and slurries were formed by
molding into gypsum molds. The dried samples were burnt in a periodic kiln with silicon
carbide heating elements at the temperature of 800–1200 °C with holding at maximum
temperature for 5–10 min. 1
Introduction Artificial ceramic binders (ACBs) are molding suspensions produced by the technology
of high-concentration ceramic binding suspensions (Pivinskii 2003) from nonplastic
materials (quartz sand, quartz glass, fire clay, etc.). The binding properties of such
suspensions are determined not by clay minerals (as in the case of traditional ceramic
slurries), but by a colloid component that forms during milling and which content can
reach up to 5%. Previous studies have shown the possible production of ACBs on the
basis of clay materials after preliminary thermal activation. The technology allows
producing high-density molding suspensions with record-low moisture content:
Wsusp = 16%, q = 1950 kg/m3 (for traditional slurry 35% and 1740 kg/m3, respec-
tively). This technology is interesting for the production of white wares, because it allows
unifying the properties of implemented raw materials and reduce the number of com-
ponents in the paste from more than 7 down to 3. This work studies the peculiarities of
ACB formation on the basis of thermally activated mixes of typical white ware com-
position: 50% of clay (kaolin from Prosyanoye deposit), 25% of fluxes (Vishnevogorsk
spar) and 25% of filling aggregates (Ziborovsk sand). © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 277–280, 2019. https://doi.org/10.1007/978-3-030-22974-0_66 I. Moreva et al. 278 3
Results and Discussion The preliminary thermal treatment of the studied raw materials allowed weakening
crystal lattices of the minerals, creating the structural non-stability due to polymorph
transformations of quartz (d/n, Å – 4.27; 3.35; 2.46; 2.29) and dehydration of clay and
hydromicaceous minerals (d/n, Å – 7.23; 3.36; 4.48; d/n, Å – 10.16; 3.30; 2.91)
(Fig. 1). After such a thermal treatment, a part of material passes into active state
(Evtushenko et al. 2007). Fig. 1. Changed phase composition of studied mixes after activation and consequent milling:
(a) initial untreated mix; (b) mix after thermal activation at 950 °C; (c) after wet milling. Fig. 1. Changed phase composition of studied mixes after activation and consequent milling:
(a) initial untreated mix; (b) mix after thermal activation at 950 °C; (c) after wet milling. The preliminary thermal activation of the material, consequent wet milling and high
density of the samples facilitates the intensification of burning, in particular, appear-
ance of mullite seeds (d/n, Å – 3.39; 5.42; 2.71; 2.55) beginning from 950 °C. In
traditional compounds, the formation of mullite begins only at 1100 °C. A part of 279 Peculiarities of Phase Formation in Artificial Ceramic Binders crystalline phases of the ACB in the white ware composition transits into the melt,
which is witnessed by the appearance of amorphous phase and decrease in the intensity
of reflections that are typical for quartz (d/n, Å – 3.35; 4.27; 2.46) and feldspars
(d/n, Å – 3.24; 3.22) (Fig. 2). Fig. 2. X-ray diagrams of burnt samples of ACB in white ware composition: (1) 950 °C;
(2) 1100 °C; (3) 1200 °C Fig. 2. X-ray diagrams of burnt samples of ACB in white ware composition: (1) 950 °C;
(2) 1100 °C; (3) 1200 °C Probably, a denser structure of the molding leads to an intense amorphization and
sintering at temperatures close to 1100 °C, while at 1200 °C close-packed structures
start to crystallize and stabilize. The high intensity of sintering and formation of a
stronger structure of ACB samples is testified by gradual increase in the coefficient of
crystallinity (Cc) with increased burning temperature: from 58.5% at 600 °C up to 91%
at 1200 °C, while for cement plant slurry it is 66.5% at the final burning temperature. The high values of Cc for samples of ACB are probably connected with more intense
processes of mullite and cristobalite formation. Evtushenko EI, Sysa OK, Moreva IYu (2007) Controlling the properties of raw materials, casting
slips, and pastes in fine-ceramic technology. Stroit Mater 8:16–17
Pivinskii YuE (2003) Highly concentrated ceramic binding suspensions (HCBS) and ceramic
castables. Stages in research and development. Refract Ind Ceram 44(3):152–160 4
Conclusions Thus, the revealed peculiarities of phase formation in ACBs of white ware composi-
tions demonstrate that the material has high reactivity in a wide temperature range. The
preliminary thermal activation of raw materials with consequent sintering of the
material facilitates earlier beginning of mullite and melt formation. The mentioned
processes are determining for the formation of the strong structure of the material and
high operation characteristics of the ceramic products. Acknowledgements. The work is realized in the framework of the Program of flagship uni-
versity development on the base of Belgorod State Technological University named after V.G. Shoukhov, using equipment of High Technology Center at BSTU named after V.G. Shoukhov. References
Evtushenko EI, Sysa OK, Moreva IYu (2007) Controlling the properties of raw materials, casting
slips, and pastes in fine-ceramic technology. Stroit Mater 8:16–17
Pivinskii YuE (2003) Highly concentrated ceramic binding suspensions (HCBS) and ceramic
castables. Stages in research and development. Refract Ind Ceram 44(3):152–160
280
I. Moreva et al. 280 I. Moreva et al. Evtushenko EI, Sysa OK, Moreva IYu (2007) Controlling the properties of raw materials, casting
slips, and pastes in fine-ceramic technology. Stroit Mater 8:16–17
Pivinskii YuE (2003) Highly concentrated ceramic binding suspensions (HCBS) and ceramic 1
Introduction One of the most promising ways to solve problem of oil pollution is to use bio-
geosorbents, which are sorbent-carriers with oil-oxidizing microorganisms immobi-
lized on their surface (Shchemelinina et al. 2017). It is possible to use zeolite and clay
raw as mineral carriers, which have high ion-exchange and sorption properties (Kotova
et al. 2017). The purpose of this work is to study sorption and destructive properties of
biogeosorbents based on clay and zeolite raw as mineral carriers for Biotrin
biopreparation. Experimental Modeling of Biogeosorbents T. Shchemelinina1, O. Kotova2, E. Anchugova1, D. Shushkov2(&),
G. Ignatyev2, and M. Markarova2 1 Laboratory of Biochemistry and Biotechnology, Institute of Biology,
Komi Science Center, Ural Branch of the Russian Academy of Sciences,
Syktyvkar, Russia
2 Laboratory of Technology of Mineral Raw, Institute of Geology named after
Academician N.P. Yushkin Komi Science Center of the Ural Branch
of the Russian Academy of Sciences, Syktyvkar, Russia
dashushkov@geo.komisc.ru 1 Laboratory of Biochemistry and Biotechnology, Institute of Biology,
Komi Science Center, Ural Branch of the Russian Academy of Sciences,
Syktyvkar, Russia
2 Laboratory of Technology of Mineral Raw, Institute of Geology named after
Academician N.P. Yushkin Komi Science Center of the Ural Branch
of the Russian Academy of Sciences, Syktyvkar, Russia
dashushkov@geo.komisc.ru Abstract. A new trend in the modeling algorithm oil sorption materials is the
adsorptive immobilization of strains of microorganisms on mineral sorbent. The
objects of study were clay and zeolite raw and biogeosorbents with oil-oxidizing
microorganisms from Biotrin preparation, immobilized on them. During our
work we modeled biogeosorbents and estimated their sorption and destructive
properties in reference to petroleum hydrocarbons. Keywords: Biogeosorbents Biotrin Petroleum hydrocarbons Sorption
Zeolites Clays © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 281–285, 2019.
https://doi.org/10.1007/978-3-030-22974-0_67 References Evtushenko EI, Sysa OK, Moreva IYu (2007) Controlling the properties of raw materials, casting
slips, and pastes in fine-ceramic technology. Stroit Mater 8:16–17
Pivinskii YuE (2003) Highly concentrated ceramic binding suspensions (HCBS) and ceramic
castables Stages in research and development Refract Ind Ceram 44(3):152 160 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches The objects of study were: 1. Mineral carriers based on analcime-bearing rocks from Koinskaya zeolite area
(Shushkov et al. 2006), clinoptilolite-bearing clays and glauconite-bearing rocks
from Chim-Loptyugskoe oil shale deposit (Saldin et al. 2013; Simakova 2016),
located in Komi Republic (Russia). For comparison, Ionsorb ™quartz-glauconite
sand from Bondarskoe deposit of Tambov region was taken as a control. 282 T. Shchemelinina et al. 2. Strains of microorganisms in the composition of Biotrin biopreparation (Conclusion
… 2017): bacteria Pseudomonas yamanorum VKM B-3033D, isolated from heavily
soiled railway bed near the city of Syktyvkar (Patent 2615458 RU); yeast Rhodo-
torula glutinis VKM Y-2998D (Patent 2658134 RU); microalgae Chlorella vulgaris
Beijer. f. globosa V. Andr. A1123. Microorganisms (cell titer 109) were cultivated
according to standard methods. Immobilization of the biopreparation on mineral
carriers was carried out in the ratio of 1 part of the biopreparation to 6 parts of the
sorbent. Initial sorbents (without Biotrin) and biogeosorbents were added to oil-
contaminated water, aerated for 4 days, and total petroleum hydrocarbons
(TPH) content in water samples, filtered initial sorbents and biogeosorbents was
measured (Method … 1998). 2. Strains of microorganisms in the composition of Biotrin biopreparation (Conclusion
… 2017): bacteria Pseudomonas yamanorum VKM B-3033D, isolated from heavily
soiled railway bed near the city of Syktyvkar (Patent 2615458 RU); yeast Rhodo-
torula glutinis VKM Y-2998D (Patent 2658134 RU); microalgae Chlorella vulgaris
Beijer. f. globosa V. Andr. A1123. Microorganisms (cell titer 109) were cultivated
according to standard methods. Immobilization of the biopreparation on mineral
carriers was carried out in the ratio of 1 part of the biopreparation to 6 parts of the
sorbent. Initial sorbents (without Biotrin) and biogeosorbents were added to oil-
contaminated water, aerated for 4 days, and total petroleum hydrocarbons
(TPH) content in water samples, filtered initial sorbents and biogeosorbents was
measured (Method … 1998). 3
Results and Discussion Norms for maximum permissible concentration (MPC) of TPH in water of fishery value
are 0.05 mg/dm3 (Order … 2016). The TPH content in the control water sample is 2.4
times higher than MPC (Table 1). g
Experiments showed that samples of the initial analcime-bearing rocks (551,
56403, 1/83) presented adsorption activity in relation to oil products. As a result of the
introduction of these samples into oil-polluted water, the content of pollutant in water
decreases 2.5–3 times in 4 days, to the MPC. When biogeosorbents are applied to
contaminated water (551-B, 56403-B, 1/83-B), the efficiency of water purification
decreases and does not reach MPC standards, which indicates decreasing sorption
properties after immobilization of microorganisms on the mineral carriers. This is
probably due to decreasing surface area of the mineral carriers covered by microor-
ganisms. During the study of initial samples and Biotrin treated samples for destructive
properties we revealed that the efficiency of oxidation of oil products in samples 551-B,
56403-B, 1/83-B increases in 4.4, 3.5 and 1.14 times, respectively. The sorption properties of clinoptilolite-bearing clays are most attractive in sample
541-31. However, taking into account a highly destructive activity of microorganisms
in 538-35-B biogeosorbent, it is preferred for remediation of oil contaminated water. Samples of initial glauconite-bearing rocks have high sorption properties (539-40,
531-56, 315-10, TG). The TPH content in the experimental water is reduced by 3.4–5
times in 4 days relative to a control sample. Biodegradation of hydrocarbons in samples of
Biotrin immobilized glauconite-bearing rocks (539-40-B, 531-56-B, 315-10-B, TG-B)
ranges from 62 to 76%. Experimental Modeling of Biogeosorbents 283 le 1. Change in the concentration of oil products in water in the presence of initial mineral
ers and biogeosorbents Table 1. 3
Results and Discussion Change in the concentration of oil products in water in the presence of initial mineral
carriers and biogeosorbents
Initial samples
TPH content*
Biotrin treated samples TPH content*
Analcime-bearing rocks
551
0.04 ± 0.014
250 ± 60
551-B
0.11 ± 0.04
57 ± 23
56403
0.046 ± 0.016
130 ± 50
56403-B
0.061 ± 0.021
37 ± 15
1/83
0.048 ± 0.017
250 ± 60
1/83-B
0.071 ± 0.025
220 ± 90
58603
0.071 ± 0.025
250 ± 60
58603-B
0.064 ± 0.022
90 ± 40
Clinoptilolite-bearing clays
538-35
0.085 ± 0.030
50 ± 20
538-35-B
0.037 ± 0.013
40 ± 16
541-31
0.035 ± 0.012
250 ± 60
541-31-B
0.058 ± 0.021
100 ± 40
Glauconite-bearing rocks
539-40
0.024 ± 0.009
58 ± 23
539-40-B
0.072 ± 0.025
20 ± 8
531-56
0.027 ± 0.009
63 ± 25
531-56-B
**
15 ± 6
315-10
0.09 ± 0.03
11 ± 4
315-10-B
0.021 ± 0.007
17 ± 7
TG
0.035 ± 0.012
90 ± 40
TG-B
0.045 ± 0.016
34 ± 14
Oil-contaminated water (control) 0.12 ± 0.041
Note: * – in the numerator, TPH content in the experimental water, mg/dm3, in the
denominator – TPH content in the initial sorbents and biogeosorbents after the experiment,
mg/g. ** – no data 4
Conclusions Our experiments resulted in modeling of biogeosorbents based on clay and zeolite raw
and oil-oxidizing microorganisms from Biotrin biopreparation immobilized on them. We determined that samples of initial sorbents showed a high adsorption activity with
respect to oil products. TPH content in water was reduced by 2.5–5 times, up to or
substantially below MPC. We revealed that microorganism cells could reduce sorption
properties of mineral carriers, at the same time providing oil destruction. Biodestruction
of oil products with biogeosorbents for 4 days was 12–77%. Acknowledgments. The authors express their gratitude to the Center for Collective Use
“Geonauka”, ecoanalytical laboratory of the Institute of Biology of the Komi Science Center UB
RAS for their assistance in analytical work. The work was carried out with the partial financial
support of UB RAS Programs (project 18-5-5-44), UMNIK project (12412GU/2017), of the State T. Shchemelinina et al. 284 task “Development of biocatalytic systems based on enzymes, microorganisms and plant cells,
their immobilized forms and associations for the processing of plant raw, production of bio-
logically active substances, biofuels, remediation of contaminated soils and wastewater treat-
ment” No. AAAA-A17-117121270025-1, “Scientific basis for effective subsoil use, development
and exploration of mineral resource base, development and implementation of innovating
technologies and economic zoning of the Timan-Nothern Ural region” No. AAAA-A17-
117121270037-4. References Certificate on the toxicological and hygienic assessment of “BIOTRIN” consortium of strains of
oil-oxidizing microorganisms. State federal enterprise for science Research center for
toxicology and hygienic regulation of biopreparations at Federal medico-biological agency,
Serpukhov, 28 September 2017 (in Russian) erpukhov, 28 September 2017 (in Russian) (1998) Method for performing measurements of the mass fraction of petroleum products in soil
samples on a Fluorat-02 analyzer. Institute of Biology of Komi Science Center of the Ural
Branch of the Russian Academy of Sciences, 16.1.21-98, Moscow, 15 p (in Russian) Patent of the Russian Federation No. 2615458 Patent of the Russian Federation No. 2658134 Order of the Ministry of Agriculture of the Russian Federation of December 13, 2016 No. 552. On approval of water quality standards for water bodies of fishery importance, including
standards for maximum permissible concentrations of harmful substances in the waters of
water bodies of fish-economic importance (in Russian) Saldin VA, Burtsev IN, Mashin DO, Shebolkin DN, Inkina NS (2013) Marking horizons in the
Upper Jurassic deposits of the Yarengsky shale region (north-east of the Russian plate), no 11,
pp 26–29. Vestnik of the Institute of Geology of the Komi Science Center UB RAS (in
Russian) Simakova YuS (2016) Features of globular layered silicates of the Chim-Loptyugsky oil shale
deposit, №9–10, pp 52–57. Vestnik of the Institute of Geology of the Komi Science
Center UB RAS (in Russian) Shushkov DA, Kotova OB, Kapitanov VM, Ignatiev AN (2006) Analcime rocks of Timan as a
promising type of minerals. Syktyvkar, 40 p (Scientific recommendations to the national
economy/Komi Science Center UB RAS, issue 123) (in Russian) Shchemelinina TN, Kotova OB, Harja M, Anchugova EM, Pelovski I, Kretesku I (2017) New
trends in the mechanisms of increasing the productivity of materials on a mineral basis, no 6,
pp 40–42. Vestnik of the Institute of Geology of the Komi Science Center UB RAS (in
Russian) Kotova OB, Harja M, Cretescu I, Noli F, Pelovski Y, Shushkov DA (2017) Zeolites in
technologies of pollution prevention and remediation of aquatic systems, no 5, pp 49–53. References Vestnik of the Institute of Geology of the Komi Science Center UB RAS Experimental Modeling of Biogeosorbents Experimental Modeling of Biogeosorbents 285 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Heating Rate and Liquid Glass Content
Influence on Cement Brick Dehydration V. Strokova(&) and D. Bondarenko Department of Material Science and Material Technology, Belgorod State
Technological University named after V.G. Shukhov, Belgorod, Russia
vvstrokova@gmail.com Department of Material Science and Material Technology, Belgorod State
Technological University named after V.G. Shukhov, Belgorod, Russia
vvstrokova@gmail.com Abstract. Peculiarities of Portland cement dehydration in different hydrate
phases with sodium water glass have been given. Three endoeffects were
determined during non-isothermal heating, connected with ettringite dehydration
and water extraction at temperature ranges 98.7–110.0 °C, calcium hydroxide
decomposition at temperature ranges 439.4–450.7 °C and secondary carbonates
decomposition at temperature ranges 657.4–669.3 °C. We experimentally
proved that the rates of dehydration of hydrated Portland cement was signifi-
cantly influenced by the liquid glass concentration. Optimum liquid glass con-
tent was grounded in the protective layer of composite finishing material,
modified with low-temperature plasma. Keywords: Plasma-chemical modification Dehydration Cement brick
Soda water glass Keywords: Plasma-chemical modification Dehydration Cement brick
Soda water glass © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 286–289, 2019.
https://doi.org/10.1007/978-3-030-22974-0_68 2
Methods and Approaches To prove the efficiency of Portland cement and liquid glass application in the protective
coating during manufacture of composite finishing material with plasma surface
treatment the samples were prepared at water\concrete ratio 0.3 from pure Portland
cement (CEM I 42,5 H) and with 5 and 10% of soda water glass (q = 1,4 g/sm3, silica
modulus 2.8) of water of mixing. After hardening at normal conditions during 28 days,
the samples were exposed to differential-thermal analysis. Plasma-chemical surface modification is done in non-isothermal conditions at
heating rate 3000 °C/min. It makes impossible to study dehydration in real conditions
of plasma heating. This process was simulated with simultaneous thermal analysis
device Netzsch STA 449 F3 Jupiter at heating rates 5 °C/min and 10 °C/min with
maximum heating rate 1000 °C. 1
Introduction Plasma-chemical modification is one of the promising technologies of creating pro-
tective–decorative coatings in the manufacture of finishing construction materials for
building and construction faces (Bondarenko et al. 2018a, b; Bessmertny et al. 2018;
Bondarenko et al. 2016; Volokitin et al. 2016). Dehydration, melt formation and
accumulation during plasma melting take second fractions, and the surface is heated up
to 2000 °C. As a result of high temperature impact hydro silicate dehydration in the
surface layers can result in micro cracking and protective-decorative coating softening,
as well as coating adhesion strength reduction and lowering of cold resisting properties. Insufficient technology elaboration on reducing heat impact consequences and
dehydration minimizing plasma melting of cement concrete does not allow wide
application of these materials on the national market. That is why the main task in
developing treatment technologies for materials based on cement matrix is composition
development for protective coating which eliminate these processes. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 286–289, 2019. https://doi.org/10.1007/978-3-030-22974-0_68 287 Heating Rate and Liquid Glass Content Influence 3
Results and Discussion The thermograph of pure hydrated Portland cement shows three endoeffects (Table 1). The first endoeffects, in the temperature range 98.7–110.0 °C in the low temperature
area, is connected with ettringite dehydration (Ca6Al2(SO4)3(OH)1226H2O) and water
extraction. Endoeffects of these two processes superimpose each other. The second is
connected with calcium hydroxide dehydration (Ca(OH)2) and happens at temperature
ranges 439.4–450.7 °C. The third endoeffects (657.4–669.3 °C) is connected with the
secondary hydro carbonates dehydration (CaCO3). Complete water extraction is at
900 °C. Table 1. Changing of endoeffects with the introduction of liquid glass and heating range 5 and
10 °C/min
Endoeffect producer
Pure
hydrated
Portland
cement
Portland
cement with
5% liquid
glass
Portland
cement with
10% liquid
glass
Heating range, °C/min
5
10
5
10
5
10
Ettringite and physically-coupled water 98.7
110.0 92.6
106.4 92.8
108.0
Calcium hydroxide
439.4 450.7 437.8 450.6 438.0 452.6
Secondary carbonate and hydro silicate 657.4 669.3 662.0 683.1 663.0 693.6 le 1. Changing of endoeffects with the introduction of liquid glass and heating range 5 and
C/min Similar results were received with the hydrated Portland cement after adding 5 and
10% liquid glass (Table 1). Similar results were received with the hydrated Portland cement after adding 5 and
10% liquid glass (Table 1). A positive effect of liquid glass adding on secondary carbonate and hydro silicate
endoeffects, which are responsible for cement brick softening and micro cracking at
higher temperature range, can be explained by effect of encapsulation of hydrate phases
with coating of liquid glass. 288 V. Strokova and D. Bondarenko Adding sodium silicate solute into Portland cement in the amount 5 and 10% takes
down mass loss (TG) in ettringite dehydration area (Fig. 1). But in high temperature
area dehydration intensity increases up to 2–3%, it is especially notable with 10%
liquid glass. Fig. 1. Dependence of water loss on time at heating range 10 °C/min: ––– Portland cement; –––
Portland cement with 5% liquid glass; ––– Portland cement with 10% liquid glass Fig. 1. Dependence of water loss on time at heating range 10 °C/min: ––– Portland cement;
Portland cement with 5% liquid glass; ––– Portland cement with 10% liquid glass The highest dehydration rate is in low temperature area (Fig. 2), which is caused by
ettringite dehydration (first climax). 4
Conclusions The influence pattern character has been determined of liquid glass content on ettringite
dehydration rate, calcium hydroxide and secondary carbonate, which is in endoeffects
shift in low temperature area to lower temperatures, and in high temperature area to the
area of high temperatures. This minimizes dehydration in low temperature area, cement
brick softening and micro cracking and as a result provides adhesion strength
improvement of protective-decorative coating with the concrete layer. Acknowledgements. The work is realized with the financial support of the Grant of the Pres-
ident for scientific schools, No. NSh-2724.2018.8, using equipment of High Technology Center
at BSTU named after V.G. Shukhov. 3
Results and Discussion The second and third climaxes are connected with
dehydration of calcium hydroxide, secondary carbonate and different hydro silicates,
are below the first climax by magnitude. Fig. 2. Dependence of water loss on time at heating rates 10 °C/min: ––– Portland cement; ––
Portland cement with 5% liquid glass; ––– Portland cement with 10% liquid glass Fig. 2. Dependence of water loss on time at heating rates 10 °C/min: ––– Portland cement; ––
Portland cement with 5% liquid glass; ––– Portland cement with 10% liquid glass Dehydration rate decrease during the first and the second climax for cement brick
with 5 and 10% of liquid glass has a significant impact on micro cracking minimization
in the surface layer of protective–decorative coating of composite finishing material at
plasma-chemical modification. Dehydration intensification can result in microcracks
increase and reduce to zero the positive influence of liquid glass in the coating. This
effect takes place at adding 10% liquid glass into Portland cement. Thus, the received 289 Heating Rate and Liquid Glass Content Influence laws analysis of mass loss and dehydration rate of the studied compositions drew to a
conclusion that the optimal is liquid glass component in the ratio 5% of mixing water. laws analysis of mass loss and dehydration rate of the studied compositions drew to a
conclusion that the optimal is liquid glass component in the ratio 5% of mixing water. References Bessmertny VS, Puchka OV, Bondarenko DO, Antropova IA, Bragina LL (2018) Plasmochem-
ical modification of wall building materials. Constr Mater Prod 1(2):11–18 Bondarenko DO, Bondarenko NI, Bessmertnyi VS, Strokova VV (2018a) Plasma-chemical
modification of concrete. Adv Eng Res 157:105–110 Bondarenko NI, Bessmertnyi VS, Borisov IN, Tymoshenko TI, Burshina NA (2016a) The
concrete with protective and decorative coverings on the basis of alyuminatny cements which
are melted off by the plasma stream. Bulletin of Belgorod State Technological University
named after V.G. Shukhov, vol 2, pp 6–12 Bondarenko NI, Bondarenko DO, Burlakov NM, Bragina LL (2018b) Investigation of influence
of plasmochemical modification on macro- and microstructure of surface layer of autoclave
wall materials. Constr Mater Prod 1(2):4–10 Volokitin O, Volokitin G, Skripnikova N, Shekhovtsov V (2016) Plasma technology for creation
of protective and decorative coatings for building materials. In: AIP conference proceedings,
vol 1698, p 070022 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Structure and Surface Reactivity Mediated
Enzymatic Performances of Clay-Based
Nanobiocatalyst S. Sun(&), K. Wang, F. Dong, B. Ma, T. Huo, Y. Zhao, H. Yu,
Y. Huang, and J. Huang Institute of Non-metallic Minerals, Department of Geological Engineering,
School of Environment and Resource, Southwest University of Science
and Technology, Mianyang, People’s Republic of China
shysun@swust.edu.cn Institute of Non-metallic Minerals, Department of Geological Engineering,
School of Environment and Resource, Southwest University of Science
and Technology, Mianyang, People’s Republic of China
shysun@swust.edu.cn Abstract. The nanobiocatalyst is an emerging innovation that synergistically
integrates advanced nanotechnology with biotechnology for improving enzyme
activity, stability, capability and engineering performances. Enzymes and clay
minerals are two important essential substances in the earth. As the important
nanostructured minerals, clay minerals have played an important role in enzyme-
related biochemical processes such as the origin and evolution of life, soil eco-
logical environment, pollution remediation and element geochemical cycling, and
even some clay minerals have intrinsic enzymatic activities. Based on the study of
clay minerals-based enzyme-like activities and enzymatic catalysis in confined
environment by clay minerals, lipase and 3-aminopropyltriethoxysilane (APTES)
functionalized montmorillonite (Mt) were selected to biomimetic construct lipase-
Mt nanobiocatalyst. Experimental results indicated that lipase-Mt activity was
40.65 U/mg, which was nearly 4-fold higher than that offree lipase under optimal
conditions. The kinetic parameters of Km and Vmax for lipase-Mt were 3.406 mM
and 312.5 mM/L min, respectively, indicating a higher affinity of the immobilized
lipase in nanobiocatalyst towards the substrate compared with free lipase. The
present study has provided a promising way for screening, optimizing and rational
design of efficient nanomineral-based enzymatic nanobiocatalyst. The present
work was supported by the National Natural Science Foundation of China
(41672039). The Longshan Academic Talent Research Support Program of the
Southwest University of Science and Technology (18LZX405). Keywords: Surface reactivity Clay minerals Enzyme
Surface modification Nanostructured minerals Nanobiocatalyst © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 290–291, 2019. https://doi.org/10.1007/978-3-030-22974-0_69 291 Structure and Surface Reactivity Mediated Enzymatic Performances Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 292–295, 2019.
https://doi.org/10.1007/978-3-030-22974-0_70 1
Introduction Structure imperfection often defines material quality and properties. The structure
perfection for layered silicates is determined by the structure of aluminosilicate layers
themselves and their positioning within crystallite. It is known that kaolinite perfect
crystals are hexagonal plane particles of the regular shape which allows free sliding
relative each other and providing plasticity, liquescence and close arrangement goods
formation. Crystallite distortion results in distortion of kaolinite plates due to lineal and
point defects. It deteriorates rheological behaviour of kaolinite suspensions, liques-
cency instability (Kukovskiy et al. 1966). It is possible to affect clay material structure by several methods (natural,
mechanical, chemical, biological and thermal) (Evtushenko et al. 2009). Hydrothermal
clay treatment can be the most effective when it is accompanied by Rehbinder effect
(adsorptive plastification) (Rebinder et al. 1972), which accelerates restructurisation
and minimizes crystal defects in clay minerals for a short period of time (Evtushenko
et al. 2006). Structural-Phase Stabilization of Clay
Materials in Hydrothermal Conditions
O. Sysa(&), E. Evtushenko, I. Moreva, and V. Loktionov O. Sysa(&), E. Evtushenko, I. Moreva, and V. Loktionov Institute of Chemical Technology, BSTU named after V.G. Shukhov,
Belgorod, Russia
sysa1975@inbox.ru Abstract. The results of thermal and X-ray phase analysis of hydrothermal
stabilized kaolin clay are given. An evaluation method has been suggested of
crystalline structure order according to the strength degree of elementary con-
tacts in clay suspensions. It has been noted that hydrothermal stabilization may
result in crystallohydrate crystalline structure perfection, in saturation, extraction
or rearrangement of water molecule in kaolinite clay structure, and influence
new phase generation during ceramic material baking. Keywords: Kaolin clay Hydrothermal processing Degree of sophistication
Crystalline Structural-phase changes Ceramics Structure and Surface Reactivity Mediated
Enzymatic Performances of Clay-Based
Nanobiocatalyst The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches X-ray phase material analysis has been done with a diffractometer “DRON-3”. XRD-
pattern was shot with CuKa filtered radiation, – radiation (Ni – filter); voltage across
the tube is 20 kw; anode tube current is 20 mA; measurement range is 10000–4000
counts per second; detectors rotation rate is 2,4 rot/min; angular mark is 10. Card index 293 Structural-Phase Stabilization of Clay Materials JCPDF was used for phase identification of elementary contacts durability formed
without particles in clay slurry. The calculation was done according to Ur’ev model
(1972, 2002) for low aggregate suspensions (Zubehin et al. 1995): JCPDF was used for phase identification of elementary contacts durability formed
without particles in clay slurry. The calculation was done according to Ur’ev model
(1972, 2002) for low aggregate suspensions (Zubehin et al. 1995): F1 ¼ cmgmind2
6;4
;
ð1Þ ð1Þ where cm is share rate corresponding to complete structural destruction, s-1;
ηmin is effective viscosity corresponding to complete structural destruction, Pa s;
d is average particle diameter, m. where cm is share rate corresponding to complete structural destruction, s-1;
ηmin is effective viscosity corresponding to complete structural destruction, Pa s;
d is average particle diameter, m. Differential-thermal analysis was done with a derivatograph OD-102. 4
Conclusions Hence it has been determined that hydrothermal treatment improves significantly sta-
bilization of clay minerals structure as well as ceramic material baking processes, that
has been proved by several research methods. Due to crystallohydrate structure change,
saturation, water molecule distribution in kaolinite structure dehydration parameters
change, processes take place which cause temperature shift of new phase chilling point
in the interval of lower temperatures. Acknowledgement. The work is realized in the framework of the Program of Flagship
University development on the base of the Belgorod State Technological University named after
V.G. Shukhov, using equipment of High Technology Center at BSTU named after V.G. Shukhov. O. Sysa et al. It has been found that elementary contact strength decreases at hydrothermal
treatment pressure and temperature increase by more than an order. Ranging by
crystalline structure defects these kaolines can be placed as follows: Zhuravliniy
Log > Glukhovetskiy > Kyshtymskiy > Prosyanovskiy and Novoselitskiy. Some differences in endo-and exothermal processes during baking in kaoline
samples depending on hydrothermal modification have been studied (Gorshkov et al. 1988). It has been determined that maximum endothermic effect of kaoline dehydration
shifts towards higher temperatures that testifies to crystalline structure sophistication. Restructurisation results in additional hydration and lessening connection energy of
crystal water in kaoline, facilitates water molecule extraction penetrating into basis of
tetrahedral kaoline layers. Thermal capacity change dependence of the studied materials during baking has been
studied. In the temperature range 950–1020 °C there are several exothermic extreme
values testifying to crystallization possibility of a variety of phase (b-crystobalite,
mullite, sillimanite, c-alumina). Structural change of the initial material causes temper-
ature shifts at the initial crystallization phase, as in clays with clearly seen crystalline
structure mullite is formed at lower temperature and in greater quantity than from dis-
ordered structure minerals. Evtushenko EI, Sysa OК (2006) Structural modification of clay raw material in hydrothermal
conditions. University news. North-Caucasian region. Technical sciences series, no 2, pp 82–
86 Evtushenko EI, Sysa OК (2006) Structural modification of clay raw material in hydrothermal
conditions. University news. North-Caucasian region. Technical sciences series, no 2, pp 82–
86
Evtushenko EI, Sysa OК, Moreva IYu, Bedina VI, Trunov Y (2009) Raw material preparation
refining during activation processes in ceramics technology. Glass Ceram 1:15–16
Gorshkov VS, Saveliev VG, Fedorov NF (1988) Physical chemistry of silicates and other high-
melting compounds. High School, Moscow
Kukovskiy Y (1966) Structure peculiarities and physic-chemical properties of clay minerals.
Naukova Dumka, Kiev Evtushenko EI, Sysa OК, Moreva IYu, Bedina VI, Trunov Y (2009) Raw material preparation
refining during activation processes in ceramics technology. Glass Ceram 1:15–16 3
Results and Discussion Kaolin clay of local deposits has been studied, which show structural changes of clay
mineral after steaming in a pressure vessel at pressure from 1 up to 4 MPa. With X-ray phase analysis we determined constant phase clay composition before
and after treatment, with significant deviations of kaolinite crystalline structure. Hydrothermal modification at saturated steam pressure up to 4 MPa drives up crys-
tallinity index according to Hinckley and intensivity of main diffractional kaolinite
reflections (Table 1), that testifies to a greater mineral structure order in the treated raw
material (Shlykov et al. 2006). But “Hinckley crystallinity index” Ch decrease depending on defects in layer
composition. This parameter is not always applicable to index correlation of index
characteristics with its real structure. Basing on the said above the authors developed an estimation method of material
structure sophistication degree according to the strength magnitude of elementary
contact, formed between particles in clay suspensions. Table 1. Change in “Hinckley crystallinity index” (Ch) of kaolinite after hydrothermal
treatment
Kaoline clay
deposits
Initial
kaoline
Kaoline after pressure
treatment
1,6 MPa
Kaoline after pressure
treatment
4,0 MPa
Zhuravliniy Log
0,20
0,22
0,46
Gluhovetskoe
0,77
0,83
1,27
Kyshtymskoye
0,55
0,47
0,61
Prosyanovskoye
0,51
0,63
0,63
Novoselitskoye
0,26
0,35
0,31 le 1. Change in “Hinckley crystallinity index” (Ch) of kaolinite after hydrothermal
ment 294 O. Sysa et al. Evtushenko EI, Sysa OК, Moreva IYu, Bedina VI, Trunov Y (2009) Raw material preparation
refining during activation processes in ceramics technology. Glass Ceram 1:15–16
Gorshkov VS, Saveliev VG, Fedorov NF (1988) Physical chemistry of silicates and other high-
melting compounds. High School, Moscow
Kukovskiy Y (1966) Structure peculiarities and physic chemical properties of clay minerals p
y
hlykov VG (2006) X-ray analysis of disperse soils mineral composition. GEOS, Moscow Rebinder PA, Ur’ev NB, Shukin Y (1972) Physic-chemical mechanics in chemical technology of
disperse systems. Theor Bases Chem Technol 6:16–24
Shlykov VG (2006) X-ray analysis of disperse soils mineral composition. GEOS, Moscow
Zubehin AP, Strakhov VI, Chekhovskiy VG (1995) Physico-chemical research methods of high-
melting non-metal and silicate materials. Synthesis, Saint-Petersburg Rebinder PA, Ur’ev NB, Shukin Y (1972) Physic-chemical mechanics in chemical technology of
disperse systems. Theor Bases Chem Technol 6:16–24
Shlykov VG (2006) X-ray analysis of disperse soils mineral composition. GEOS, Moscow Rebinder PA, Ur’ev NB, Shukin Y (1972) Physic-chemical mechanics in chemical technology of
disperse systems. Theor Bases Chem Technol 6:16–24 Zubehin AP, Strakhov VI, Chekhovskiy VG (1995) Physico-chemical research methods of high-
melting non-metal and silicate materials. Synthesis, Saint-Petersburg 1
Introduction Bismuth oxide is a widely used bismuth compound due to its chemical stability, low
toxicity, and its unique physical and chemical properties (Gulbin et al. 2014). Its
sufficiently high density (8.9 g/cm3) allows to use it in the synthesis of “heavy”
materials employed in the protection from gamma rays. As it is known, in this case, the
leading role in characterizing the protective properties is played by the density of the
material. In the way of exercising high structural properties, the most efficient were the
ceramic matrices (Correya et al. 2018). This paper studies the phase transformations of the radiation protection materials
based on the high-alumina ACBs, bismuth oxide, and aluminum oxide sintered at
different temperatures. S. Yashkina(&), V. Doroganov, E. Evtushenko, O. Gavshina,
and E. Sysa Department of Technology of Glass and Ceramics, Belgorod State Technological
University named after V.G. Shukhov, Belgorod, Russia
yashkina_asp@mail.ru Abstract. XRF method was used to study the mineralogical compounds of the
radiation protection ceramic materials based on a high-alumina binder and a
“heavy” aggregate, bismuth oxide. The content of Bi2O3 in the test samples was
kept in the range of 38.5–75 wt%. Along with the bismuth oxide, the aggregate
was aluminum oxide (Al2O3). The binder synthesis followed the principle of
obtaining ceramic concretes based on artificial ceramic binders (ACB). The paper establishes the specifics of sintering of the composites under study,
fired under different temperatures. Keywords: Bismuth oxide Artificial ceramic binders X-ray fluorescence
Radiation protection ceramic composites © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 296–299, 2019.
https://doi.org/10.1007/978-3-030-22974-0_71 References Evtushenko EI, Sysa OК (2006) Structural modification of clay raw material in hydrothermal
conditions. University news. North-Caucasian region. Technical sciences series, no 2, pp 82–
86 Evtushenko EI, Sysa OК (2006) Structural modification of clay raw material in hydrothermal
conditions. University news. North-Caucasian region. Technical sciences series, no 2, pp 82–
86 Evtushenko EI, Sysa OК, Moreva IYu, Bedina VI, Trunov Y (2009) Raw material preparation
refining during activation processes in ceramics technology. Glass Ceram 1:15–16 Gorshkov VS, Saveliev VG, Fedorov NF (1988) Physical chemistry of silicates and other high-
melting compounds. High School, Moscow Kukovskiy Y (1966) Structure peculiarities and physic-chemical properties of clay minerals. Naukova Dumka, Kiev Kukovskiy Y (1966) Structure peculiarities and physic-chemical properties of clay minerals. Naukova Dumka, Kiev Structural-Phase Stabilization of Clay Materials 295 Zubehin AP, Strakhov VI, Chekhovskiy VG (1995) Physico-chemical research methods of high-
melting non-metal and silicate materials. Synthesis, Saint-Petersburg Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches Obtaining the artificial ceramic binder (ACB) was carried out through the technique of
wet grinding in a periodic action ball mill with a stepwise loading of material. This
principle of obtaining ceramic concretes allows to obtain matrices with the desired
phase composition, improved physical and mathematical properties and ensures the
broad opportunities for using various aggregates. 297 Phase Changes in Radiation Protection Composite Materials The suspension based on high-alumina grog possesses the thixotropic dilatant flow
(the destruction of the initial thixotropic structure and the following dilatant formation
of structure). The bismuth oxide powder used as aggregate represents spherical parti-
cles sized no more than 35 µm. The source materials were mixed in the proportions presented in Table 1. Then, the
technique of static pressing was used with the specific pressure of 100 MPa to form the
bar. After drying, the samples were subjected to burning within the interval of 200° to
1000 °C with holding at the maximum temperature for one hour. Table 1. Compositions of radiation protection materials
Number of the compound Content, wt%
ACB Al2O3 Bi2O3
1
38.5
23.0
38.5
2
19.0
23.0
58.0
3
–
25.0
75.0 Table 1. Compositions of radiation protection materials The X-ray fluorescence analysis was performed using the DRON-3 diffractometer. XRD patterns were recorded using the CuKa radiation (Ni filter); tube voltage: 20 kV;
tube anode current: 20 mA; measuring range: 10000–4000 PPS; detector turn rate:
2.4 rpm; elevation angle: 10. The JCPDF database was used for identification. 4
Conclusions This paper studied the mineralogical compounds of the radiation protection ceramic
materials based on a high-alumina ACB and bismuth oxide. It was identified that the
higher the content of Bi2O3 in the source composition, the more intensive is the process
of mullite decomposition. This way, at the content of Bi2O3 38,5%, mullite perseveres
up to the temperature of 900 °C, and if bismuth oxide is increased to 58%, mullite
disintegrates completely at the temperature of 700 °C, which is 200 °C lower than the
previous composition. Acknowledgement. The work is realized in the framework of the Program of Flagship
University development on the base of the Belgorod State Technological University named after
V.G. Shukhov, using the equipment of High Technology Center at BSTU named after V.G. Shukhov. Phase Changes in Radiation Protection Composite Materials Phase Changes in Radiation Protection Composite Materials It is possible to assume that compositions No. 1–2 show the formation of bismuth
silicates as a result of mullite decomposition and binding of SiO2 into the bismuth
silicates, while in the composition No. 3 the silicates form due to the interaction of
Bi2O3 and the nanosilica introduced as binder. 3
Results and Discussion The analysis of the transformations of the samples with the admixture Bi2O3 at varied
temperatures (Figs. 1, 2 and 3) indicates that heating within the interval of 100–500 °C,
all compounds are mineralogically constant and exhibit phases of mullite, corundum,
and Bi2O3. When increasing the burning temperature to 600 °C, starts the formation of bismuth
silicates 2Bi2O33SiO2 and 12Bi2O32SiO2. Further increase of the burning temperature
to 700 °C leads to a more intensive formation of bismuth silicates, indicated by the
increase in the intensity of reflections for these compounds. In the process, the entire
Bi2O3 turns into silicates, while the interplanar reflections of bismuth oxide totally
disappear. There occurs a sharp decrease in the intensity of mullite peaks, while in the
composition No. 3 this compound is totally absent. When the temperature is increased up to 800 °C and further, all compositions dis-
play further formation of bismuth silicates, and in the compositions No. 2–3 there were
identified two phases of a-Al2O3 и 12Bi2O32SiO2, while in the composition No. 1, in
addition to the indicated two, there also forms 2Bi2O33SiO2 (Figs. 1, 2 and 3). S. Yashkina et al. 298 Fig. 1. X-ray patterns of the samples of the composition of radiation protection composites
No. 1 Fig. 1. X-ray patterns of the samples of the composition of radiation protection composites
No. 1 No. 1 Fig. 2. X-ray patterns of the samples of the composition of radiation protection composites
No. 2 Fig. 2. X-ray patterns of the samples of the composition of radiation protection composites
No. 2 Fig. 3. X-ray patterns of the samples of the composition of radiation protection composites
No. 3 Fig. 3. X-ray patterns of the samples of the composition of radiation protection composites
No. 3 299 Development of Technology for Anti-corrosion
Glass Enamel Coatings for Oil Pipelines E. Yatsenko(&), A. Ryabova, and L. Klimova South Russian State Polytechnical University (NPI), Novocherkassk, Russia
e_yatsenko@mail.ru Abstract. Among anticorrosive coatings for steel products, glass-enamel glass
is the most reliable and versatile, based on aluminoborosilicate glasses of the
SiO2-Al2O3-B2O3-R2O system. This anti-corrosion coating allows to increase
the chemical resistance of the internal surface of pipelines to various groups of
reagents. Therefore, in the course of the study, a previously developed com-
position was modified by introducing oxides SrO, ZrO2, CaO, MoO3, Li2O and
their acid and alkalinity of enamel frits and coatings based on them and it was
found that the addition of strontium and zirconium oxide in the amount of 2%
was optimal. Keywords: Oil pipelines Anti-corrosion coatings Glass-enamel coatings
Steel protection Chemical resistance Acid and alkali resistance © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 300–303, 2019.
https://doi.org/10.1007/978-3-030-22974-0_72 References Correya AA, Mathew S, Nampoori VPN, Mujeeb A (2018) Structural and optical character-
ization of hexagonal nanocrystalline bismuth-bismuth oxide core-shell structures synthesized
at low temperature. Optik 175:930–935 Gulbin VN, Kolpakov NS, Polivkin VV (2014) Radio - i radiatcionno-zashchitnye kompozit-
cionnye materialy s nanostrukturnymi napolniteliami [Radiation protection composite
materials with nanostructure aggregates]. VSTU Bull 23:43–51 Gulbin VN, Kolpakov NS, Polivkin VV (2014) Radio - i radiatcionno-zashchitnye kompozit-
cionnye materialy s nanostrukturnymi napolniteliami [Radiation protection composite
materials with nanostructure aggregates]. VSTU Bull 23:43–51 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches The technology of enameling the internal surface of pipelines includes the following
technological stages: preliminary annealing of steel pipes at a temperature of *750 °C
in order to remove contaminants and decarburize the outer layers of steel; mechanical
preparation of the inner surface of pipes using shot blasting using single or multiple
shot blasting units to remove scale and roughen for better adhesion to the glass-enamel
coating; preparation of enamel slip suspension on the basis of finely ground glass
granulate; application of slip suspension on the inner surface of a horizontal pipe by
sprinkling using an impeller; drying at a temperature of 100–120 °C and induction
roasting at a temperature of 860–880 °C. The aim of the work was to develop the composition and technology of applying a
glass-enamel single-layer coating for medium-carbon steels, which has high rates of
chemical and abrasive resistance and is capable of forming a defect-free smooth coating
on the steel surface. To solve this problem, the aluminoborosilicate system SiO2-
Al2O3-B2O3-R2O was chosen as the most acceptable in the single-layer enamelling
technology, which was modified by introducing compounds such as SrO, ZrO2, CaO,
MoO3, Li2O, in order to improve chemical resistance and defect-free formation in the
form of through pores. The introduction of enamels strontium, calcium and zirconium
oxides that are insoluble in oxides helps to reduce the leaching of alkali and alkaline
earth cations when exposed to acid coating. Lithium oxide together introduced with
oxides of sodium and potassium contributes to the chemical resistance of enamel
coatings due to the manifestation of polyalkalnochnogo and polycationic effects. The
amount of additives introduced into the charge was 2 wt.%, Since the introduction of
additives less than 1% slightly affects the properties of enamel coatings, and more than
2% can greatly affect the change in the technological properties of the melt. Next, the compounded mixtures were boiled at a temperature of 1350 °C for 1 h in
an electric oven in alundum crucibles, subjected to wet granulation and applied in the
form of finely ground enamel slip to samples of 32G2S steel. After drying and firing at
a temperature of 850 °C, the resulting enamel coatings were tested to study the effect of
modifying oxides on the structure and properties of enamels. 1
Introduction Currently, the oil industry of the Russian Federation is developing rapidly and effi-
ciently. However, corrosion of equipment and facilities in the oil and gas industry is
one of the main reasons for reducing their performance, causing huge economic losses
and environmental damage. Presently, protective coatings (bitumen, epoxy, poly-
urethane, etc.) have become widely used to protect oil pipelines, among which glass-
enamel coatings with high chemical resistance and thermomechanical properties, in
particular, heat resistance, reaching 500 C, occupy a special place. Due to the fact that
the silicate-enamel coating for steel pipelines is exposed to an aggressive environment
containing hydrocarbons and formation water, in which chlorides, sulfates and organic
acids are present, as well as up to 10% hydrogen sulfide and 10% carbon dioxide. The
coating composition was based on an acid-resistant glass composition (Ryabova et al. 2018) with a high content of quartz and low boric anhydride and alkaline oxides, which
will improve the chemical resistance of enamels and extend the range of their roasting
(Yatsenko et al. 2018). Also, when choosing the type of protective coating, the following factors should be
taken into account: operating conditions, composition of the transported medium,
temperature and pressure in the system, speed and nature of the flow movement,
presence of abrasive particles in the fluid flow, composition and properties of associ-
ated petroleum gas (APG), presence of asphalt-resin-paraffin deposits (AFS), the
manifestation of the life of microorganisms. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 300–303, 2019. https://doi.org/10.1007/978-3-030-22974-0_72 301 Development of Technology for Anti-corrosion Glass Enamel Coatings Therefore, the purpose of these studies was the development of anticorrosive glass-
enamel coatings to protect steel pipelines from medium carbon steel 32G2S and the
study of factors affecting the process of their defect-free formation and technical and
operational properties. 2
Methods and Approaches To assess the corrosion resistance, tests were carried out to determine the acid
resistance characterized by weight loss after exposure to 20% boiling hydrochloric acid
and alkali resistance - weight loss after exposure to 8% sodium hydroxide solution. For
frits, the tests were carried out by the grain method, and for enamel coatings, the impact
on their surface. 302 E. Yatsenko et al. 3
Results and Discussion For the developed modified frits and enamel coatings, chemical resistance to various
groups of reagents for weight loss after boiling in acid and alkaline solutions was
studied. The test results are presented in Table 1. able 1. Indicators of the properties of modified frits and enamel coatings depending on th
mposition Table 1. Indicators of the properties of modified frits and enamel coatings depending on the
composition composition
Name of enamel Modifying additive Chemical resistance
Frites, %
Coatings, mg/cm2
20%-HCl 8%-NaOH 20%-HCl 8%-NaOH
10-0
Without additives
0,38
0,83
0,22
0,78
10-1
SrO
0,32
0,99
0,16
0,45
10-2
ZrO2
0,40
1,20
0,20
0,60
10-3
CaO
0,38
0,75
0,21
0,63
10-4
MoO3
0,54
0,98
0,22
0,68
10-5
Li2O
0,42
0,87
0,23
0,63 The results obtained allow us to conclude that the introduction of modifying
additives into the glass matrix composition has a significant impact on anti-corrosion
properties. For all compositions, the mass loss of frits after boiling is quite significant, since the
specific surface of glass powders is much larger than the surface of the burned enamel
coating. However, the composition of frits modified by strontium and zirconium oxides
is characterized by less mass loss, which is caused by their positive effect on the
increase in the packing density of the structural amorphous network, due to the large
radius of these ions. Molybdenum oxide has almost no effect on chemical resistance,
but it has a positive effect on the continuity of the coating contributing to a smoother
formation and the absence of coating defects, due to a decrease in the surface tension of
the enamel melt. Calcium oxide in such an amount does not affect the chemical
resistance of frits and coatings. Lithium oxide increases chemical resistance and frit and
coatings, although it is alkaline in itself, but it has an inhibitory effect due to the
presence of several alkali cations (Na2O and K2O). 4
Conclusions As a result of the research, the composition and technology of producing anticorrosive
glass-enamel coatings for the internal protection of steel pipelines based on the SiO2-
Al2O3-B2O3-R2O aluminoborosilicate system has been developed. The effect of vari-
ous modifiers of the vitreous matrix on the acid and alkali resistance was studied, and it
was found that the strontium and zirconium oxides in an amount of 2% are optimal. 303 Development of Technology for Anti-corrosion Glass Enamel Coatings Development of Technology for Anti-corrosion Glass Enamel Coatings Acknowledgements. The work was done with the financial support of the Russian Science
Foundation under the agreement No. 18-19-00455 “Development of technology for the inte-
grated protection of pipelines for oil and gas operated in the Far East of Russia” (headed by
Yatsenko E. A.). Ryabova AV, Yatsenko EA, Klimova LV, Goltsman BM, Fanda AY (2018) Protection of steel
pipelines with glass-enamel coatings based on silica-containing raw materials of the far east of
Russia. Int. J. Mech. Eng. Technol. 9(10):769–774 g
Yatsenko EA et al (2018) Investigation of chemical processes that ensure the adhesion strength of
glass-enamel coating with steel pipelines. Butlerov Commun 56(11): 122–127 Ryabova AV, Yatsenko EA, Klimova LV, Goltsman BM, Fanda AY (2018) Protection of steel
pipelines with glass-enamel coatings based on silica-containing raw materials of the far east of
Russia. Int. J. Mech. Eng. Technol. 9(10):769–774
Yatsenko EA et al (2018) Investigation of chemical processes that ensure the adhesion strength of
glass-enamel coating with steel pipelines Butlerov Commun 56(11): 122 127 References Ryabova AV, Yatsenko EA, Klimova LV, Goltsman BM, Fanda AY (2018) Protection of steel
pipelines with glass-enamel coatings based on silica-containing raw materials of the far east of
Russia. Int. J. Mech. Eng. Technol. 9(10):769–774 g
Yatsenko EA et al (2018) Investigation of chemical processes that ensure the adhesion strength of
glass-enamel coating with steel pipelines. Butlerov Commun 56(11): 122–127 Yatsenko EA et al (2018) Investigation of chemical processes that ensure the adhesion strength of
glass-enamel coating with steel pipelines. Butlerov Commun 56(11): 122–127 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Optimization of Formulations of Cement
Composites Modified by Calcined Clay Raw
Material for Energy Efficient Building
Constructions A. Balykov(&), T. Nizina, V. Volodin, and D. Korovkin
Department of Building Structures, Ogarev Mordovia State University,
Saransk, Russia
artbalrun@yandex.ru A. Balykov(&), T. Nizina, V. Volodin, and D. Korovkin Department of Building Structures, Ogarev Mordovia State University,
Saransk, Russia
artbalrun@yandex.ru Abstract. The paper presents the results of the study of the influence of for-
mulation and process parameters of dehydrated raw material preparation based
on polymineral clay rocks of the Republic of Mordovia used as independent
mineral additives to cement composites. The possibility of increasing the studied
physical and mechanical parameters of composites by optimizing the mode of
clay raw material calcination and the content of the developed modifier is
shown. Keywords: Cement composite Dehydration Mineral additive Clay Keywords: Cement composite Dehydration Mineral additive Clay © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 307–310, 2019.
https://doi.org/10.1007/978-3-030-22974-0_73 2
Methods and Approaches Clay from Staroshaygovsky deposit (The Republic of Mordovia) was selected as a raw
material for mineral additive development. To carry out the experimental studies, a plan
was prepared, which includes 15 experiments allowing variation of the temperature and
duration of calcination at three levels (400, 600 and 800 °C and 2, 3 and 4 h,
respectively), and the content of mineral additive based on thermally-activated clay in
the composition of cement composites on five levels – 2, 6, 10, 14 and 18% of the
weight of Portland cement. Also, the additive-free composition (No. 16) was studied in
addition to the 15 formulations included in the main block of the experiment plan. Manufacture of cement compositions was carried out at a fixed water-solid ratio of 0.3. The calcined clays were ground in a planetary mill for 1 h. The resulting fine powder
was introduced into the cement binder based on Portland cement CEM I 42.5 N
produced by Serebryakovcement PJSC. According to the results of the study, opti-
mization of the modified cement binders was carried out and the most effective cal-
cination modes were determined. 1
Introduction Currently, Portland cement is the main binder in the construction industry. Introduction
of fine-grained mineral additives of natural and man-made origin to Portland cement in
order to improve the indicators of its physical and mechanical properties and partially
replace clinker is one of the effective ways to ensure sustainable development in terms
of resource conservation. In recent years, such mineral additives as microsilica and
metakaolin have been increasingly used for more rational use of Portland cement and
ensure the required level of cement composites characteristics. These modifiers help to
increase the density of cement stone by controlling its phase composition and porosity,
thereby allowing improvement of physical, mechanical and operational properties of
cement composites at reduced cement consumption (Kirsanova et al. 2015; Nizina et al. 2017; Dvorkin et al. 2015). However, the resources of the above additives do not meet the increasing needs of
the construction industry. In this regard, researchers face the challenge of expanding the
resource base for the production of mineral additives using available natural raw
materials. One of the most promising in this respect are calcined clay rocks (Schulze
et al. 2015). At the same time, according to the studies (Rakhimov et al. 2017; Fernandez
et al. 2011) results, it was found that kaolinite, montmorillonite and muscovite/illite
clays have the highest pozzolanic activity after heat treatment. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 307–310, 2019. https://doi.org/10.1007/978-3-030-22974-0_73 308 A. Balykov et al. The territory of Russia is rich in various types of clays. Ordinary (low-melting) clay
in Russia is produced almost everywhere. For example, in the territory of the Republic
of Mordovia there are more than fifty deposits of clay rocks, which allows classifying
the development of active mineral additives based on clay raw materials as promising
task of the construction industry, the solution of which a number of economic, tech-
nological and environmental problems of the cement industry both in the region and in
country as a whole. 2
Methods and Approaches Rational compositions were determined according to
the analysis of an experimental statistical model describing the change in compression
resistance of cement composites based on modified calcined clay raw materials: y ¼ 67; 29 þ 3; 23 x1 þ 0; 18 x2 3; 99 x3 þ 1; 36 x1 x2
0; 81 x1 x3 1; 38 x2 x3 þ 0; 31 x1 x2 x3 7; 55 x2
1
4; 35 x2
2 þ 4; 48 x2
3 0; 91 x2
1 x2 0; 56 x1 x2
2
0; 49 x2
1 x3 2; 19 x1 x2 x3
ð
Þ2
ð1Þ y ¼ 67; 29 þ 3; 23 x1 þ 0; 18 x2 3; 99 x3 þ 1; 36 x1 x2
0; 81 x1 x3 1; 38 x2 x3 þ 0; 31 x1 x2 x3 7; 55 x2
1
4; 35 x2
2 þ 4; 48 x2
3 0; 91 x2
1 x2 0; 56 x1 x2
2
0; 49 x2
1 x3 2; 19 x1 x2 x3
ð
Þ2
ð1Þ ð1Þ Identification of compromise solutions optimal areas for each factor separately was
carried out using frequency ranges, which is one of the most descriptive ways to
graphically represent the random variable probability density (Lyashenko et al. 2017). References Dvorkin LI, Zhitkovsky VV, Dvorkin OL, Razumovsky AR (2015) Metakaolin is effective
mineral additive for concrete. Concrete Technol 9–10(110–111):21–24 Fernandez R, Martizena F, Scrivener KL (2011) The origin of the pozzolanic activity of calcined
clay minerals: a comparison between kaolinite, illite and montmorrilonite. Cement Concrete
Res 41:113–122 Kirsanova AA, Ionov YV, Orlova AA, Kramar LY (2015) Features of hydration and hardening
of cement concretes with additives modifiers containing metakaolin. Cement Appl 2:130–135
L
h
k
TV V
k
VA (2017) C
iti
fild
th d l
i Kirsanova AA, Ionov YV, Orlova AA, Kramar LY (2015) Features of hydration and hardening
of cement concretes with additives modifiers containing metakaolin. Cement Appl 2:130–135 Lyashenko TV, Voznesensky VA (2017) Composition-process fields methodology in compu-
tational building materials science. Astroprint, Odessa Lyashenko TV, Voznesensky VA (2017) Composition-process fields methodology in compu-
tational building materials science. Astroprint, Odessa Lyashenko TV, Voznesensky VA (2017) Composition-pr
tational building materials science. Astroprint, Odessa Nizina TA, Balykov AS, Volodin VV, Korovkin DI (2017) Fiber fine-grained concretes with poly-
functional modifying additives. Mag Civil Eng 4:73–83. https://doi.org/10.18720/MCE.72.9 Nizina TA, Balykov AS, Volodin VV, Korovkin DI (2017) Fiber fine-grained concretes with poly-
functional modifying additives. Mag Civil Eng 4:73–83. https://doi.org/10.18720/MCE.72.9 Rakhimov RZ, Rakhimova NR, Gaifullin AR, Morozov VP (2017) Properties of Portland cement
paste incorporated with loamy clay. Geosyst Eng 6:318–325 Rakhimov RZ, Rakhimova NR, Gaifullin AR, Morozov VP (2017) Properties of Portland cement
paste incorporated with loamy clay. Geosyst Eng 6:318–325 Schulze SE, Pierkes R, Rickert J (2015) Optimization of cements with calcined clays as
supplementary cementations materials. In: Proceedings of XIV international congress on the
chemistry of cement, Beijing, China Schulze SE, Pierkes R, Rickert J (2015) Optimization of cements with calcined clays as
supplementary cementations materials. In: Proceedings of XIV international congress on the
chemistry of cement, Beijing, China Schulze SE, Pierkes R, Rickert J (2015) Optimization of cements with calcined clays as
supplementary cementations materials. In: Proceedings of XIV international congress on the
chemistry of cement, Beijing, China Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. 4
Conclusions According to the results of the study, optimal formulation and process principles for the
production of mineral additive based on clay raw materials were determined, which
allow increasing compression resistance of modified cement composites in comparison
with the additive-free composition. The most effective additives were obtained at
calcination time from to 3 to 3.6 h at clay calcination temperature 640 720 °C. The data obtained indicate the prospects and relevance of the development of
concrete with modifying additives based on thermally-activated polymineral clays,
which allows expanding the range of modified cement composites produced today due
to better use of local mineral resources base. Acknowledgements. The reported study was funded by RFBR and Government of the Republic
of Mordovia according to the research project №18-43-130008. 3
Results and Discussion According to the results of the conducted study, it was determined that a number of
modified cement composites can achieve compression resistance equal to 70 80 MPa,
which is comparable with the control composition No. 16 (Fig. 1). The highest strength
characteristics were achieved in compositions 2, 4, 6 and 13 at a content of calcined clay
from 2 to 6% of the cement weight. Optimization of Formulations of Cement Composites 309 Fig. 1. Compression resistance of modified cement composites at the age of 7 and 28 days Fig. 1. Compression resistance of modified cement composites at the age of 7 and 28 days а)
b)
Fig. 2. The range of distribution of the compression resistance of modified cement composites
at the age of 28 days: a – from calcination temperature, b – from calcination duration а) b) Fig. 2. The range of distribution of the compression resistance of modified cement composites
at the age of 28 days: a – from calcination temperature, b – from calcination duration The analysis of the ES model (1) based on frequency ranges (Fig. 2) showed that
the compression resistance corresponding to the control composition can be provided
for cement composites with a mineral additive at any studied temperature level and
duration of calcination of the clay raw material. At the same time, for the accepted
temperature and time intervals of the mineral additive calcination, the total proportion
of compositions with enhanced or corresponding to the control composite character-
istics varies from 22 to 41% depending on the duration and from 11 to 45% depending
on the temperature of calcination. It was found that an increase in clay calcination time
from 2 to 3 4 h leads to an expansion of the relative values range of modified cement
composites strength characteristics from 77.5 115 to 62.5 130%. Increasing the
temperature of calcined clay rocks calcination from 400 to 720 °C allows changing the
limit (achievable) range of compression resistance from 62.5 107.5 to 85 130%, a
further increase in temperature leads to a certain decrease in the boundary values of the
relative strength indicator to 77.5 (lower boundary) and 122.5% (upper boundary),
respectively. 310 A. Balykov et al. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 311–312, 2019.
https://doi.org/10.1007/978-3-030-22974-0_74 1
Introduction Santa Maria, the oldest and most oriental island from the archipelago, is very weathered,
as consequence of intense volcanic activity alternated with sea level alterations and
intense erosion episodes. There are residual and sedimentary clayey deposits in several
parts of the island, one of which known as “Red Desert”, corresponding to Feteiras
Formation. In addition to the previous, Almagreira Formation is other important deposit
of the island. These two in particular were more studied. The abundance of raw mate-
rials, gave this island an ancient tradition concerning exploitation of clays for pottery,
being known as the “mother island” of clay. From this island, during centuries, white
clays were extracted and exported to provide other islands. Recently a pottery oven was
discovery “in the middle” of Vila do Porto and it date from the 18th century. Santa Maria Clays as Ceramic Raw Materials Â. Cerqueira1(&), C. Sequeira1, D. Terroso1, S. Moutinho1, C. Costa1,
and F. Rocha1,2 1 GeoBioTec, Geosciences Department, University of Aveiro,
3810-193 Aveiro, Portugal
angelamcerqueira@ua.pt
2 RISCO, Civil Engineering Department, University of Aveiro,
3810-193 Aveiro, Portugal Abstract. There are evidences and records concerning clay exploitation for
pottery in the island of Santa Maria (Azores, Portugal) in the past. Nowadays
this activity is almost extinct but this is the only island in the archipelago with
abundant residual and sedimentary clay deposits. To evaluate the applicability of
clays from this island for modern ceramics, a campaign made in May 2017
allowed collecting twenty samples in several outcrops all over the island. All
samples were subsequently analyzed in terms of granulometry, mineralogy,
chemical composition and physical properties. Results revealed to be interesting,
namely concerning mineralogical composition, where phyllosilicates such as
Kaolinite are in high percentages. Granulometry also revealed that most part of
the samples is composed by fine grain size particles (<63 µm), which can be a
good indicator of the existence of resources in great quantity. Keywords: Santa Maria Island Pottery Physical characterization
Potentialities Keywords: Santa Maria Island Pottery Physical characterization
Potentialities References The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches Chemical composition was assessed by X-ray fluorescence, qualitative and semi-
quantitative mineralogical analyses were carried out by X-ray diffraction and crystal-
lochemistry analyses were carried on a Scanning Electron Microscope (SEM) Hitachi Â. Cerqueira et al. 312 SU70 with Energy Dispersive X-Ray Spectroscopy (EDS) Brucker QUANTAX 400. Viscosity was assessed with a Haake Viscotester iQ. Plasticity Index was also com-
puted from Atterberg Limits determination. SU70 with Energy Dispersive X-Ray Spectroscopy (EDS) Brucker QUANTAX 400. Viscosity was assessed with a Haake Viscotester iQ. Plasticity Index was also com-
puted from Atterberg Limits determination. Alkaline Activation of Rammed Earth
Material – “New Generation of Adobes”
C. Costa1(&), D. Arduin1, C. Sequeira1, D. Terroso1, S. Moutinho2,
Â. Cerqueira1, A. Velosa1,2, and F. Rocha1,2 C. Costa1(&), D. Arduin1, C. Sequeira1, D. Terroso1, S. Moutinho2,
Â. Cerqueira1, A. Velosa1,2, and F. Rocha1,2 1 GeoBioTec, Geosciences Department, University of Aveiro,
3810-193 Aveiro, Portugal
cristianacosta@ua.pt
2 RISCO, Civil Engineering Department, University of Aveiro,
3810-193 Aveiro, Portugal Abstract. Adobe is an extremely simple form of earth construction and with
this technique the shrinkage associated with the construction of large structures
is avoided. In Portugal, earthen materials have been used in load-bearing walls
in the form of adobe or rammed earth for the construction of buildings especially
in the southern and central coast. Most conventional consolidation treatments
used in the past have not succeeded in providing a long-term solution because
they did not tackle the main cause of degradation, the expansion and contraction
of constituent clay minerals in response to humidity changes. Clay swelling
could be reduced significantly by transforming clay minerals into non-
expandable binding materials with cementing capacity using alkaline activa-
tion. in this study it was being developed adobes with water, and alkaline
activated with NaOH and KOH. The obtained results allowed to conclude that
the adobes with NaOH and KOH have an increase of its properties. Keywords: Rammed-earth Adobe construction Alkaline activation
“New generation of adobes” Keywords: Rammed-earth Adobe construction Alkaline activation
“New generation of adobes” 3
Results and Conclusions Results are very positive since most part of samples are rich on phyllosilicates (69% to
98%, being kaolinite the most common), fine-grained, and showing adequate plasticity
and viscosity. Geological formations, in particular Feteiras and Almagreira Formations,
outcrops on a large part of the island, the residual ones showing always intensive
alteration, therefore assuring the existence of good reserves. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches The chemical composition of diatomite samples was assessed by X-ray fluorescence
(XRF), qualitative mineralogical analyses were carried out by X- ray diffraction
(XRD). Compressive strength tests use a universal mechanical compression testing
machine (Shimadzu Autograph AG 25 TA). 1
Introduction The use of adobe construction in the Aveiro district reflected the properties of the
existing raw materials available to be applied, namely sand, clay sediments and soils
and lime (Silveira et al. 2012), and there is an evident heterogeneity of the adobes
linked to the geographic distribution of the available resources. In Aveiro there was a
semi-industrial production of adobe, some small companies employing ‘adobeiros’, for
the manufacture of blocks of adobe, along with a domestic self-production (Costa et al. 2016). Most conventional consolidation treatments used in the past have not succeeded
in providing a long-term solution because they did not tackle the main cause of
degradation, the expansion and contraction of constituent clay minerals in response to
humidity changes. Clay swelling could be reduced significantly by transforming clay
minerals into non-expandable binding materials with cementing capacity using alkaline
activation. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 313–314, 2019. https://doi.org/10.1007/978-3-030-22974-0_75 314 C. Costa et al. 3
Results and Conclusions Mineralogically, samples are composed by quartz, feldspars and phyllosilicates,
however, there is an evidence of the presence of amorphous alumino-silicate phases. Samples with higher mechanical strength are associated with higher specific surface
areas, namely, those with NaOH. The alkaline activation promotes the increase of
mechanical resistance. With the exception of one sample, all the samples present
promising mechanical resistances for their application in rehabilitation works on adobe
buildings, since these do not require high resistance, even for the sake of compatibility. Costa CS, Rocha F, Velosa AL (2016) Sustainability in earthen heritage conservation.
Sustainable use of traditional geomaterials in construction practice. Geol Soc London, Spec
Publ 416. http://doi.org/10.1144/SP416.22
Silveira D Varum H Costa A Martins T Pereira H Almeida J (2012) Mechanical properties of Silveira D, Varum H, Costa A, Martins T, Pereira H, Almeida J (2012) Mechanical properties of
adobe bricks in ancient constructions. Constr Build Mater 28:36–44 Costa CS, Rocha F, Velosa AL (2016) Sustainability in earthen heritage conservation.
Sustainable use of traditional geomaterials in construction practice. Geol Soc London, Spec
Publ 416. http://doi.org/10.1144/SP416.22
Silveira D, Varum H, Costa A, Martins T, Pereira H, Almeida J (2012) Mechanical properties of
adobe bricks in ancient constructions. Constr Build Mater 28:36–44 © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 315–318, 2019.
https://doi.org/10.1007/978-3-030-22974-0_76 M. Elistratkin1(&), V. Lesovik1, N. Chernysheva1, E. Glagolev2,
and P. Hardaev3 1 Department of Building Materials, Products and Designs,
Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
elistratkin.my@bstu.ru, mr.elistratkin@yandex.ru
2 Department of Construction and Municipal Economy,
Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
3 Department Industrial and Civil Engineering,
East Siberian State University of Technology and Management,
Ulan-Ude, Russia 1 Department of Building Materials, Products and Designs,
Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
elistratkin.my@bstu.ru, mr.elistratkin@yandex.ru
2 Department of Construction and Municipal Economy,
Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
3 Department Industrial and Civil Engineering,
East Siberian State University of Technology and Management,
Ulan-Ude, Russia Abstract. One of new and perspective lines of development in the field of
construction technologies is the integration of additive production elements. The
laboratory study of these issues revealed critical challenges that slightly slowed
down the introduction of construction printing in daily construction practice. One of such problems is a big difference of structurization conditions of additive
composites produced via traditional methods. The paper provides the analysis of
factors exerting negative influence on structurization and considers the possi-
bilities of solving such challenges. Keywords: Construction printing Mixing ratio Structurization of composites Composite binding agents Mineral additives References Costa CS, Rocha F, Velosa AL (2016) Sustainability in earthen heritage conservation. Sustainable use of traditional geomaterials in construction practice. Geol Soc London, Spec
Publ 416. http://doi.org/10.1144/SP416.22 Silveira D, Varum H, Costa A, Martins T, Pereira H, Almeida J (2012) Mechanical properties of
adobe bricks in ancient constructions. Constr Build Mater 28:36–44 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. M. Elistratkin et al. Another problem, which is currently being tackled by scientists, is the production
of efficient mixes for printing, which is considered controversial from the perspective
of the traditional concrete technology and with regard to its sufficient properties. It is
impossible to ensure competitiveness of construction additive technologies in relation
to traditionally applied methods without the development of new principles of their
creation that would integrate classic approaches and the latest achievements in con-
struction materials science (Yi et al. 2017). 2
Methods and Approaches The study used the traditional cement-sand mortar at the ratio of 1:4 at W/C = 0.45…
0.5 as an extrusion printing mix. The following refer to special properties of mortar,
which make its different from standard mixes: – ability to easily pass through the extruder and a 20 20 mm nozzle without losing
uniformity and sticking to walls; – ability to easily pass through the extruder and a 20 20 mm nozzle without losing
uniformity and sticking to walls; – ability to hold its shape after extrusion and resist loading of at least 5 layers placed
on top without intermediate curing. – ability to hold its shape after extrusion and resist loading of at least 5 layers placed
on top without intermediate curing. The specified qualities were obtained due to combination of two Russian additives
characterized by availability, low cost and good technological effectiveness. The used
additives allow receiving various mixes for construction printing at the C:S ratio from
1:3 to 1:5 that preserve special properties within 25…30 min. 1
Introduction One of new and quite perspective development areas of construction technologies is the
integration of additive production elements. The active development of construction
printing launched a decade ago allowed creating some concepts demonstrating only a
mere part of its potential and drawing public attention and investments to its development
(De Schutter et al. 2018). However, it also caused some critical problems that slightly
slowed down the introduction of construction printing in daily construction practice. First of all, such problems include structural reinforcement. In the most cases
traditional reinforcement methods (frames, rods, grids) do not correspond to the ide-
ology of additive production, which implies that the construction robot installs the
building structure without too much human involvement and use of various additional
technical tools. Various methods of dispersed (Christ 2015) and textile reinforcement
(Lesovik et al. 2017) may possibly solve the matter, and in the long-term perspective
this problem may be solved by making the construction printer place or ‘raise’ the
reinforcement according to the design project. 316 M. Elistratkin et al. 3
Results and Discussion The printing with the developed mixes on a laboratory unit (Fig. 1) sets the task to
obtain the flattest surface without post-processing. Fig. 1. Printing with developed mixes on a laboratory unit Fig. 1. Printing with developed mixes on a laboratory unit Table 1 shows the strength properties of the mix during processing in various
conditions. The preparation of a molding compound is critical since this stage ensures
the formation of traditional and special properties. Structurization of Composites When Using 3D-Additive Technologies 317 According to our experience in the study of construction printing, it is more
preferable for industrial facilities to ensure continuous supply of the mix into a small
bin feeder installed on a forming device. According to our experience in the study of construction printing, it is more
preferable for industrial facilities to ensure continuous supply of the mix into a small
bin feeder installed on a forming device. Table 1. Strength of mixes (MPa) depending on curing conditions Table 1. Strength of mixes (MPa) depending on curing conditions
Composition
1 day 3 day 7 day 28 day
1:3 (in water)
3.7
8.2
12.6
17.9
1:3 (in insulation) 4.2
8.2
11.3
14.3
1:3 (in air)
4
6.9
8.1
8.5
1:4 (in water)
2.9
6.3
9.4
12.9
1:4 (in insulation) 3
6.8
10.5
15.2
1:4 (in air)
3
5.4
6.6
6.8 The stage when the mix passes through the extruder is followed by its additional
mixing, decrease in viscosity, and utilization of some amount of air. Printing with slight
premolding of lower layers with the newly placed ones ensures the formation of
stronger contact between them compared to free mix outflow, but at the same time
imposes increased requirements on the ability of a mix to keep its shape after extrusion
(Secrieru et al. 2017). The third stage is characterized by the largest duration and ensures the formation of
final indicators of the additive structure. The curing conditions of composites in the printed structure significantly (for the
worse) differ from traditional methods of concrete works (Zharikov et al. 2018). The
wall assemblies made via outline printing have small effective sectional area at their
quite big surface area, which leads to their fast dehydration. Table 1 shows the
influence of curing conditions on strength accumulation velocity. 3
Results and Discussion It is less likely possible to create favorable conditions for structurization of com-
posites due to external moistening, therefore it is possible to define solution to this
problem: reduction of a binder’s demand in water and the maximum increase in early
strength during the period until the main amount of liquid has not evaporated yet. As it was noted in some works (Chernysheva et al. 2013; Sumskoy et al. 2018;
Lesovik et al. 2014; Kuprina et al. 2014; Elistratkin et al. 2018), a good solution in such
cases may be the replacement of portland cement with composite binding agents that
contain mineral additives with developed microporosity (zeolites, utilization products of
some ceramic construction materials). Such products are able to create some moisture
stock, which maintains hydration for some time needed for the material to become strong. 4
Conclusions Alongside with various processing technologies, quite often complicating the design of
a construction printer, the essential positive effect may be achieved by using composite
binding agents with required properties: required rheology, ability to quickly gain
strength during solidification in the conditions of fast dehydration, reduced or zero
shrinkage. Such measures will provide for a simpler solution to the task of creating 318 M. Elistratkin et al. favorable conditions for composite structurization in construction printing thus estab-
lishing a self-sufficient system not too much dependent on external factors. favorable conditions for composite structurization in construction printing thus estab-
lishing a self-sufficient system not too much dependent on external factors. Acknowledgements. The study is implemented in the framework of the RFBR according to the
research project No. 18-03-00352, using equipment of High Technology Center at BSTU named
after V.G. Shukhov. References Chernysheva NV, Ageeva MS, Elyan I, Drebezgova MYu (2013) The effect of mineral additives
of different genesis on the microstructure of the gypsum-cement stone. Bull. BSTU named
after V.G. Shukhov 4:12–18 Christ S (2015) Fiber reinforcement during 3D printing. Mater Lett 139:165–168 De Schutter G, Lesage K, Mechtcherine V, Nerella VN, Habert G, Agusti-Juan, I (2018) Vision
of 3D printing with concrete – technical, economic and environmental potentials. Cement
Concrete Res Elistratkin MYu, Minakova AV, Jamil AN, Kukovitsky VV, Eleyan Issa Jamal Issa (2018)
Composite binders for finishing compositions. Constr Mater Products 1(2):37–44 Kuprina AA, Lesovik VS, Zagorodnyk LH, Elistratkin MY (2014) Anisotropy of materials
properties of natural and man-triggered origin. Res J Appl Sci 9(11):816–819 Lesovik VS, Chulkova IL, Zagordnyuk LK, Volodchenko AA, Yurievich PD (2014) The role of
the law of affinity structures in the construction material science by performance of the
restoration works. Res J Appl Sci 9(12):1100–1105 Lesovik VS, Popov DYu, Glagolev ES (2017) Textile-concrete is an effective reinforced
composite of the future. Constr Mater 3:81–84 Secrieru E, Fataei S, SchröflC, Mechtcherine V (2017) Study on concrete pumpability combining
different laboratory tools and linkage to rheology. Constr Build Mater 144:451–461 Sumskoy DA, Zagorodnyuk LKh, Zhernovskiy IV (2018) Features of the formation of crystalline
neoplasms in astringent compositions depending on the technology of their preparation. Bull BSTU named after V.G. Shukhov 6:71–78 Yi WDT, Biranchi P, Suvash CP, Nisar ANM, Ming JT, Kah FL (2017) 3D printing trends in
building and construction industry: a review. Virtual Phys Prototyping 12(3):261–276 Zharikov IS, Laketich A, Laketich N (2018) Impact of concrete quality works on concrete
strength of monolithic constructions. Constr Mater Products 1(1):51–58 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction The scientists and technologists should develop a technology of new generation of
cellular concrete with a higher strength and frost-resistance at a low average density. Influence of Flow Blowing Agent
on the Properties of Aerated Concrete
Variable Density and Strength V. Galdina(&), E. Gurova, P. Deryabin, M. Rashchupkina,
and I. Chulkova Department of Building Structures, Ogarev Mordovia State University,
Saransk, Russia
example@yandex.ru Abstract. We presented the effect of gasifier flow on gas release kinetics
and basic properties of aerated concrete with variable traverse density and
strength. We found optimal consumption of gasifier for manufacturing of aer-
ated concrete with variatropic properties from expanded clay sand, wherein we
obtained aerated concrete with a strength of 1.42 MPa at an average density of
414 kg/m3. Keywords: Cellular concrete Aerated concrete Blowing consumption
Bloating Gas release kinetics © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 319–322, 2019.
https://doi.org/10.1007/978-3-030-22974-0_77 3
Results and Discussion The formation of a cellular structure and gas-concrete products in an individual mold
was carried out as follows: gas-concrete mix was prepared, poured into a mold closed
by a cover with a circular hole in the center of the mold (The method…, 2015). The essence of formation of a cellular structure of concrete of variable traverse
density and durability is as follows. Hydrogen releases when an alkaline component
binds with aluminum, which results in blowing of viscoplastic mass. The blowing mix,
having reached the internal surface of the cover with a circular hole, meets a barrier on
the way and swells up on the way of the weakest resistance (through the hole), and on
the periphery of the product there is a self-consolidation of aerated concrete due to
overpressure. As a result, the pressing gas with more than 0.1 kgf/cm2 and presence of
the closed cover surface at the time of a blowing results in less dense and more porous
products in the mid-range of the composite and denser, stronger – on the periphery. The known formulations in production of gas-concrete products by traditional
technology, in particular a gas developing agent consumption, are not absolutely
correct for manufacturing of products in the closed mold since the raised gas devel-
oping agent consumption is necessary for effective self-consolidation of exemplars. In
this regard it is necessary to reveal its influence on a kinetics of gas emission of the mix
and main properties of aerated concrete. V/T = 0.5 was applied to aerated concrete on the basis of expanded clay sand with
mix spread at Southard viscosimeter 30 cm. Gazobetolyuks gas paste and earlier
chosen area of the closed cover surface with the round section equal to 71% was
applied as gas developing agent. The mix was poured up to 70% of the mold height. The nature of a flatulence of the mix up to the cover in the period of time from 5 to
20 min at all gas-concrete mix was identical, different only by the amount of gas
developing agent. Process of an aerogenesis in all mixtures practically finishes in 20–
30 min (Fig. 1). The highest limit of compression strength is observed for samples produced at a
gasifier flow 1000 g per m3 of the mix. 2
Methods and Approaches The production of concrete products in a closed mold can be related to new methods. Works by Chernov and Zavadsky are important at the present stage of production of
aerated concrete. Their studies are mainly based on preparation of gas concrete mix in
the mold with a hollow cap (without holes), or small holes in side and top faces of the
mold (Chernov et al. 1983; Chernov 2003; Zavadsky 2005; Zavadsky 2001; Chernov
2002; Zavadsky, Kosach 1999). The properties of aerated concrete with variable traverse density and strength of
products are influenced by the following factors: area of surface of a cover; fluidity of
mix; consumption and type of blowing agent; fill level of mix in the mold; type, flow
rate and surface area of silica component and binder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 319–322, 2019. https://doi.org/10.1007/978-3-030-22974-0_77 V. Galdina et al. 320 3
Results and Discussion As a result of the chemical reaction the released
hydrogen swells the mix which reaches the edge of the mold, where it meets an
obstacle of cover and is pressed on the way of the weakest resistance through a circular
hole in the cover, whereby mostly form crusts and increase strength of samples during
compression. When the flow reduces to 800 g the strength is reduced on average by
30%. This is a result of low flow of the gasifier, and the mix during swelling is not
sufficient for self-sealing along the periphery, which is confirmed by gas release
kinetics, which reaches the inner surface of the cover only in 20–22 min and the final
height of swelling 35%. Since one of the main objectives is not only to obtain aerated
concrete with a relatively high compressive strength, but also with reduced average
density index, the optimum flow rate is from 1100 to 1300 g per m3. 321 Influence of Flow Blowing Agent Fig. 1. Kinetics of gas emission of mix on the basis of expanded sand Fig. 1. Kinetics of gas emission of mix on the basis of expanded sand Aerated concrete, prepared in the mold with a circular hole in the cover on
expanded clay aggregate with the strength 1.42 MPa at an average density 414 kg/m3
(Fig. 2), was obtained at a flow rate equal to 1,200 g of blowing agent per m3 of mix,
which corresponds to D500 brand strength. The gassing process also takes about 25–
30 min, after which the height of the crusts is 50–54% (Fig. 2). Fig. 2. The effect of the gasifier consumption on the average density and strength of aerated
concrete based on expanded clay sand Fig. 2. The effect of the gasifier consumption on the average density and strength of aerated
concrete based on expanded clay sand V. Galdina et al. 322 4
Conclusions We determined the optimal flow of gas agent on the basis of expanded clay sand equal
to 1100–1300 g per m3, at which the aerated concrete with a durability of 1.42 MPa at
an average density of 414 kg/m3 that corresponds to D500 brand was produced. At the
same time the process of aerogenesis continues 25–30 min, after which the height of
top crust is 50–54%. Advantage of technology of aerated concrete with variatropic properties is a high
stability of density of cellular concrete depending only on the accuracy of the gravimetric
feeders measuring components. At the same time not only density, but also durability,
deformability, heat conductivity and other properties are stabilized. The increased dis-
persion of operational parameters, inherent to all cellular concretes, is excluded. Development of new and combination of already known methods of pore making
in mass and also application of processing methods for products with variatropic
properties at formation allow producing one and two-layer wall products of various
configuration, sizes and required heat-shielding and operational properties. Structuring Features of Mixed Cements
on the Basis of Technogenic Products M. Garkavi1, A. Artamonov1, E. Kolodezhnaya2(&), A. Pursheva1,
and M. Akhmetzyanova1 1 JSC “Ural-Omega”, Magnitogorsk, Russia
2 Department of Mining and Ecology, Institute of Comprehensive Exploitation
of Mineral Resources, Russian Academy of Sciences, Moscow, Russia
gsm@uralomega.ru Abstract. The structure of mixed cements with mineral additives of different
nature enables to divide them into two types: with single-stage and multi-stage
structure formation. Characterization of mixed cements is determined, mainly,
by nature of the mineral admixture. The index of multi-stage structure formation
of mixed cements has been suggested. This index is the value of instantaneous
power of structure formation Wη. It has been shown that in case of the other
equal conditions the nature of structure formation is determined by the ratio of
the components and by milling method of the mixed cement. As a result of
analysis of numerous experimental data it has been detected, that the strength of
stone, which is formed during hardening of mixed cements, has an extreme
nature depending on the content of the mineral admixture. Analysis of the
thermodynamic stability of the structure of the stone, which is formed in the
process of hardening of mixed cements, allows to divide mixed cements into
three groups and identify field of their rational use. Keywords: Mixed cement Structure formation
Instantaneous power of structure formation © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 323–326, 2019.
https://doi.org/10.1007/978-3-030-22974-0_78 References Chernov AN (2003) Auto-frettage in aerated concrete technology. Constr Mater 11:22–23 Chernov AN (2002) Cellular concretes. Publishing house of SUSU, Chelyabinsk, 111 p Chernov AN, Kozhevnikova LP, Khmelev SV, Tsarkov VV, Danilyuk MA, Moiseev EI,
Stepanova ZA (1983) Technology of cellular concrete products with compacted surface layer. Constr Mater 8:12–13 he method of producing aerated concrete and the raw mix for its preparation. Patent of th The method of producing aerated concrete and the raw mix for its preparation. Patent of the
Russian Federation P. Deryabin. No. 2560009 of July 20 2015 Russian Federation P. Deryabin. No. 2560009 of July 20 2015 Zavadsky VF (1999) An integrated approach to solving the problem of thermal protection of
walls of heated buildings. Constr Mater 2:7–8 Zavadsky VF (2001) Variants of wall constructions with the use of effective heaters -
Novosibirsk: NGASU, 52 p Zavadsky VF, Kosach AF (2005) Wall materials and products. SibADI Publishing House, Omsk,
254 p Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Research Methods Portland cement clinker, granulated blast-furnace slag, steelmaking slag and flyash
were used for mixed cement production of different mixture ratio. All components of mixed cements were grinded in VIS mills not only for fineness
needed but also for their mechanoactivation. The real size of BET surface area is
10000…10400 sq. m/kg, which is an indication of high defect structure and related
concentration of surface specific sites. 1
Introduction Technogenic products have a low hydraulic activity. Therefore, the main condition is to
increase their reaction capacity in making the mixed binders, which is achieved by
increasing the degree of fineness and concentration of blemishes. Milling is an energy intensive process operation. So, the energy reduction is
achieved by using modern fineness of grind, of which vertical shaft impacted mill is the
most effective. The feature of VIS mill is their high-power rating (more than 10 kW/kg). This
allows using them in mechanoactivation processing when solid materials increase
capacity of reaction. The simultaneous grinding and mechanoactivation in VIS mills provide to diversify
types of cements (Garkavi et al. 2013). M. Garkavi et al. 324 3
Results and Discussion During hardening of mixed binding systems several chemical reactions of hydration are
connected with various hydraulic activity of their components. Each chemical reaction
of hydration, which occurs in a mixed binding system is interconnected with one process
of structure formation and affects its development. The character of structure formation
of mixed cements with mineral additives of different nature enables to divide them into
two types: with a single-stage or multi-stage structure formation (Garkavy 2005). The
nature of the mineral admixture shows that mixed cements belong to one or another type. The multi-stage structure formation means the cyclicity in occurrence of certain
structural conditions of mixed binding system. The number of stages of structure
formation is determined by the composition of mixed cement such as coagulation or
coagulation-condensation contacts. Prerequisite for the development of multi-stage structure formation is a significant
distinction of the apparent activation energies of hydrate-formation of mixed binding
components (Garkavy 2005). Values of apparent activation energy for components of mixed binding is shown in
Table 1. Table 1. Activation energy of hydration of the components of the mixed cem
Component of mixed cement Activation energy of hydration for
the period of, J/mol
Induction Acceleration Damping
Portland cement clinker
15703
14610
23736
Flyash
47837
17288
31563
Granulated blast-furnace slag 15668
14703
23898
Steelmaking slag
43675
19953
35595 Table 1. Activation energy of hydration of the components of the mixed cements The contribution of components of the mixed cement to the formation of inter-
particle contacts can be estimated by the quantity of instantaneous power of structure
formation Wη. This value is the product of the rate constant of structure formation
process kη and the value of the apparent activation energy of this process Eη, i.e. Wη = kη* Eη (Garkavy 2005). By the synchronous development of the processes of hydrate and structure for-
mation, in other words, by the one-stage structure formation the correlation is present: Structuring Features of Mixed Cements Structuring Features of Mixed Cements 325 Wgmix\Wgcl þ Wgmin:ad
ð1Þ ð1Þ where Wgmix, Wgcl and Wgmin:ad – instantaneous power of structure formation of
the mixed cement, clinker component and mineral admixture, respectively. Consequently, by one-stage structure formation both components of the mixed
cement are involved in the formation of structure, but the predominant influence has a
component, which possesses a high instantaneous power structure formation. 3
Results and Discussion In case of
breach of synchronicity of these processes, that is, by the multi-stage structure for-
mation, the relation is implemented: Wgmix ¼ Wgcl þ Wgmin:ad
ð2Þ ð2Þ It means that in the structure formation components of the mixed cement are
consistently involved, the component with the greater instantaneous power of structure
formation contributes the most. The correlation (2) which takes into account the mass fraction u of the mineral
admixture in the mixed cement has the form: Wgmix ¼ 1 u
ð
Þ Wgcl þ u Wgmin:ad
ð3Þ ð3Þ where u - the mass fraction of mineral admixture in the mixed cement. From the formula (3) follows, that ceteris paribus, the nature of structure formation
is determined by the ratio of the components of the mixed cement. According to the
result of the experimental research, the multi-stage structure formation is typical of the
mixed cements based on steelmaking slags (u > 0.3) and fly ash (u > 0.5) (Garkavi
et al. 2010). Thus, the Eq. (3), along with the ratio of the quantity of activation energy of
hydrate formation components of the mixed cement can be an indicator of multi-stage
structure formation in the binder systems. It is obvious that in the process of hardening
of mixed cements with the diverse character of structure formation, intermediate and
final structural states with diverse thermodynamic stability are formed. It predetermines
their various physical-mechanical and operational characteristics and creates the pre-
requisites for the creation of rational compositions of mixed cements which have
desired properties. As a result of numerous experimental data, it has been detected (Garkavi et al. 2010; Garkavi et al. 2013), that the strength of stone which is formed in the process of
hardening of mixed binders, has an extreme character which depends on the mass
fraction of the mineral admixture in the mixed cement. The extreme dependences, which are shown in Fig. 1, are approximated by the
empirical equation: Rmix ¼ Rcl expðb / þ c /2Þ;
ð4Þ ð4Þ where Rmix – activity of the mixed cement Rcl - activity of Portland cement clinker; b,
c - constants depending on the nature and dispersion of the mineral admixture; u - the
mass fraction of mineral admixture of mixed cement. 326 M. Garkavi et al. 4
Conclusion In agreement with the nature of hardening of mixed cements, they can be divided into
three groups with the following rational fields of application: Group 1 – mixed cements which contain the mineral admixtures u ucr are
characterized by single-stage structure formation, they comply with Eq. (1). The mixed
cements of this group are substitutes of cements with no additives. Group 2 - mixed cements with multi-stage structure formation, which satisfy the
relation (3). These cements should be used for the manufacture of concretes, which hard-
ened by the heat treatment, as well as for slow-hardening concretes for massive structures. Group 3 – cements content a high mass fraction of mineral admixture (80%). These
cements form the weak strength structure of hardening with low thermodynamic sta-
bility. The cements of this group should be used for the production of concrete and
low-grade concretes and solutions. The offered classification of mixed cements allows to identify their rational com-
positions, on the basis of the properties of mineral admixture and applications. 3
Results and Discussion From the correlation (4) follows that the value b/c characterizes the mass fraction of
mineral admixture of mixed cement u cr, in which its activity equal to the activity of
the Portland cement with no additives. According to the Eq. (3), in the mixed cement
the conditions for the development of multi-stage formation are created when the
content of mineral admixture is more than u cr. 1
Introduction Any waste can be considered as secondary material resources, which may be fully or
partially (as additives) used in production (Klassen et al. 2003). In terms of their
physical and chemical properties, slags are similar to igneous rocks used in the pro-
duction of construction materials. Being exposed to high-temperature treatment and
containing basic calcium silicates, they considerably reduce fuel consumption during
clinker burning (Vvedensky 1978; Kopeliovich et al. 1998). Use of Slags in the Production of Portland
Cement Clinker V. Konovalov(&), A. Fedorov, and A. Goncharov Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
konovalov52@mail.ru Abstract. The use of technogenic raw materials as input products in the pro-
duction of portland cement provides for considerable reduction of energy con-
sumption during clinker burning. The study reveals the mineral formation
features caused by the change of the liquid phase composition and crystalliza-
tion of silicate phases. The use of slags in raw mixes increased the mechanical
strength of cements by over 50%. Keywords: Slag Cement Clinker phases Microstructure Keywords: Slag Cement Clinker phases Microstructure Keywords: Slag Cement Clinker phases Microstructure © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 327–330, 2019.
https://doi.org/10.1007/978-3-030-22974-0_79 References Garkavy MS (2005) Thermodynamic analysis of structural transformations in the binder systems. MSTU, Magnitogorsk, 243 p Garkavy MS (2005) Thermodynamic analysis of structural transformations in the binder systems. MSTU, Magnitogorsk, 243 p
Garkavi MS, Hripacheva IS (2010) Optimization of mixed binders with the use of dump
steelmaking slag. Build Mater 2:56
Garkavi MS, Hripacheva IS, Artamonov AV (2013) Centrifugal impact grinding cements. Cement Appl 4:106–109 Garkavy MS (2005) Thermodynamic analysis of structural transformations in the binder systems. MSTU, Magnitogorsk, 243 p
G k
i MS H i
h
IS (2010) O i i
i
f
i
d bi d
i h h
f d Garkavi MS, Hripacheva IS, Artamonov AV (2013) Centrifugal impact grinding cements. Cement Appl 4:106–109 Garkavi MS, Hripacheva IS, Artamonov AV (2013) Centrifugal impact grinding cements. Cement Appl 4:106–109 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches The study included the methods of chemical analysis of input products and clinkers
carried out according to GOST 5382-93. The X-ray phase analysis was performed via
the powder diffraction technique using DRON-3 M. The polished sections were studied
in reflected light using a universal polarizing microscope NU-2 by Karl Zeiss Jena. The
etching of polished sections was carried out via the universal etching agent, i.e. M.I. Strelkov’s reagent. The thermal test was carried out using the scanning calorimeter
STA 449 F1 Jupiter® by NETZSCH in inert media. Thin slag structures were studied
on a scanning electron microscope MIRA3 TESCAN. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 327–330, 2019. https://doi.org/10.1007/978-3-030-22974-0_79 328 V. Konovalov et al. Table 2. Raw mixing ratio Table 2. Raw mixing ratio Raw mix, No. Limestone Clay Blast furnace slag Open-hearth slag
1
75%
22%
0%
3%
2
61%
0% 33%
6% The clear exo-effect is observed in a clay-based raw mix (No. 1) at 1227 and 1256 °C
caused by the formation of belite phase, as well as endothermal melting effects at 1288
and 1300 °C. According to (Kougiya, Ugolkov 1981), the exotherm of belite mass
crystallization at higher temperatures improves the synthesis of alite and its formation in
a fine-crystalline state. Within sintered materials cooled at 1250 °C the crystallization of phase C3A and
C4AF is recorded, the reflection intensity of aluminate phase considerably increases with
temperature rise. When clay is replaced with slag in a mix the exothermal processes
characterizing the formation of silicate phases are weakly expressed within the range of
1000 and 1200 °C. The melting in these mixes is recorded at 1259 and 1308 °C. 1
2
Fig. 2. Micrographs of polished section of sample clinkers: identifications are given in Table 1 ig. 2. Micrographs of polished section of sample clinkers: identifications are given in Table Within sintered materials №2 cooled at 1250 °C the aluminate phase prevails thus
leading to the appearance of ferrous phases – C6A2F and C4AF with temperature rise. The basic clinker fusion is formed at 1327 °C. These differences in A-F formation are
clear with the increase of slag concentration in raw mixes. The gehlenite and mayenite
is observed at 1200 °C in clay-containing mixes along with belite phase, which is not
observed in slag-containing samples. The introduction of slag intensifies the formation
of belite phase at early burning stages. Some changes in the composition of a ‘liquid’
phase, which increase its temperature and molten viscosity with the increase of slag
composition in a raw mix, also affect the features of crystallization of clinker phases. Figure 2 shows micrographs of polished sections of sample clinkers, which
demonstrates the crystallization difference of alite phase. Clinkers from clay-based raw mixes have clear monadoblastic texture. Clinkers
from slag-based raw mixes are different in terms of the number of alite growths. The strength of cements was defined in small samples from cement paste (1:0) with
water-cement ratio of 0.28. The results given in Table 3 show that the use of slags in
raw materials instead of clay positively affects the activity of clinkers. 3
Results and Discussion Besides others, the LLC South-Ural Mining and Processing Company utilizes smelter
slags as input products. The chemical composition of slags and raw materials given in
Table 1 indicates a possibility of replacing the natural constituent, i.e. clay, and slightly
reducing the consumption of a carbonate component. Table 1. Chemical composition of input products, %. Table 1. Chemical composition of input products, %. Raw mix components CaO
Al2O3 Fe2O3 SiO2
MgO
PPP
Blast furnace slag
39.70
8.80
2.09
39.08
4.08
0.17
Open-hearth slag
32.36
3.96 21.20
19.76 11.18
3.61
Limestone
52.12
0.24
0.18
1.53
0.48 43.11
Clay
7.57 12.8
6.32
50.45
3.97 12.80 According to X-ray phase analysis, the phase composition of dump blast furnace non-
granulated and granulated slag differs only in the intensity of new crystal growths. The
main identifiable minerals include gehlenite, akermanit, quartz and melilite minerals. The DTA method of a dump blast furnace slag in the range of 600–700 °C
demonstrates a slight vague exo-effect caused by ferrous iron oxidation. The same
temperature range indicates the decomposition of a secondary calcium carbonate fol-
lowed by the insignificant loss of a sample mass. At 840–900 °C the glass phase is
crystallized with further heat release. Fig. 1. Microstructure of dump blast furnace slag Fig. 1. Microstructure of dump blast furnace slag The process of devitrification for the blast furnace granulated slag has more expressed
exothermic maximum, which is caused by high concentration of a glass phase. The process of devitrification for the blast furnace granulated slag has more expressed
exothermic maximum, which is caused by high concentration of a glass phase. The open-hearth slag acts as a correcting ferrous additive, besides hematite and
magnetite is rich in calcium ferrite, monticellite, diopside and magnesium oxide in the
form of a periclase. Figure 1 shows the structure of dump slags representing the conglomerate of
crystalline phases and melting particles. The phase formation analysis was carried out
in raw mixes having similar chemical composition with various ratio of clay and slag
components (Table 2). Use of Slags in the Production of Portland Cement Clinker Use of Slags in the Production of Portland Cement Clinker 329 Table 2. Raw mixing ratio The strength
improvement within a 28-day interval at full replacement of a clay component with slag
made 62%. The heat burning input of limestone-slag mixes may be reduced by over
0.85 mJ/t. 330 V. Konovalov et al. Table 3. Cement stone strength, W/C = 0.28
Mix No., Identifications
are given in Table 1
Tensile strength,
MPa, days
2
7
28
1
7
14
32
2
30
37
52 Table 3. Cement stone strength, W/C = 0.28 4
Conclusions The integral analysis of raw materials and calcined products indicates the possibility of
full replacement of clay in a raw mix for the production of portland cement clinker. This contributes to the improvement of qualitative parameters of a calcined product and
to the reduction of its production cost. Acknowledgements. The study is implemented in the framework of the Flagship University
Development Program at Belgorod State Technological University named after V.G. Shukhov,
using equipment of High Technology Center at BSTU named after V.G. Shukhov. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 331–334, 2019.
https://doi.org/10.1007/978-3-030-22974-0_80 1
Introduction The application of colloid and nano-sized silicate and aluminosilicate components for
the synthesis of effective binding systems is one of the most attractive directions in the
science of construction materials (Vivian et al. 2017; Dmitrieva et al. 2018; Sobolev
2016). The earlier studies (Shekhovtsova et al. 2018; Galindo Izquierdo et al. 2009) dis-
cussed various factors influencing the ability of alkali activated cements to form the
aluminosilicate structures from the anthropogenic aluminosilicates, in particular fly-ash
(Kozhukhova et al. 2018; Wang et al. 2018). Geopolymerization and Structure Formation
in Alkali Activated Aluminosilicates
with Different Crystallinity Degree N. Kozhukhova1(&), V. Strokova1, I. Zhernovsky1, and K. Sobolev2 1 Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
kozhuhovanata@yandex.ru
2 University of Wisconsin-Milwaukee, Milwaukee, USA Abstract. The work presents the results of grain-size analysis of alkali-
activated aluminosilicate suspensions with different crystallinity degree. It is
found that the crystallinity degree of aluminosilicate is inversely proportional to
its solvability in strong alkaline substance. The mechanism of geopolymeric
system formation during the geopolymerization process has been suggested. Keywords: Aluminosilicates Crystallinity degree Structure formation
Geopolymerization Keywords: Aluminosilicates Crystallinity degree Structure formation
Geopolymerization References Klassen VK, Borisov IN, Klassen AN, Manuylov VE (2003) Features of mineral formation in
slag-containing raw mixes of various basicity. Bull High Educ Inst Constr Ser 7:56–58 Kopeliovich VM, Zdorov AI, Zlatkovsky AB (1998) Utilization of industrial wastes in cement
production. Cement 3:174 Kougiya MV, Ugolkov VL (1981) Differential thermal analysis of portland cement raw mixes. Cement 11:19–21 Vvedensky VG (1978) Environmental and economic efficiency of waste utilization. Complex
Use Mineral Raw Mater 3:59 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. N. Kozhukhova et al. These suspensions were mixed for three days (72 h) using a LS-110 mixing device. The specific surface and the average size of grains of aluminosilicate powders was
performed using a laser analyzer ANALYSETTE 22 NanoTec plus. 2
Materials and Methods To estimate the viability of the research hypothesis three types of natural aluminosil-
icates with a different crystallinity degree were used: – Obsidian - effusive rock of an acidic composition and amorphous structure; – Obsidian - effusive rock of an acidic composition and amorphous structure; – Pearlite - effusive rock of an acidic composition and crypto-crystalline structu – Crouan – intrusive compound acidic rock with a hollow crystalline structure. To prepare alkaline silicate suspensions the equal volumes (50 g) corresponding to
each sample of milled aluminosilicate material were placed into glass bottles and mixed
with 50% NaOH water solution. 332 N. Kozhukhova et al. 4
Conclusion The crystallinity degree of aluminosilicates is inversely proportional to the reactivity in
alkali systems, which is controlled by the solubility degree in highly alkali medium. Based on this observation, it was suggested that the geopolimerization scheme in the
system «gel layer – unreacted grain of aluminosilicate component» occurs during the
alkaline activation. Acknowledgements. The work has been fulfilled within the project Federal Target Program of
Research and Development on “Priority Development Fields of science and technology sector in
Russia for 2014–2020”, unique project number is RFMEFI58317X0063. 3
Results and Discussions The structure formation mechanism in the system “gel layer – unreacted grain of
aluminosilicate component” during the alkali activation. The lower aluminosilicate crystallinity degree results in a high intensity of the
dissolution of aluminosilicate particles and a thicker surface gel layer based on the
newly formed compound. 3
Results and Discussions This report is based on the hypothesis that during the alkaline activation of alumi-
nosilicate particles the dissolving process is gradual, starting with the dissolution of
surface layers. As the result, the alkali aluminosilicate gel is formed which acts as a
binding base for further geopolymerization. At the same time, the aluminosilicate
component crystallinity degree influences its solubility in the alkaline medium. To test the research hypothesis the average size of the aluminosilicate particles
average size was determined as well as the specific area in the initial condition and also
after the alkaline activation (Table 1). Table 1. The particle size and specific area of alumosilicate powders after the a Table 1. The particle size and specific area of alumosilicate powders after the alkaline activation
№Aluminosilicate
type
Average particle size,
lm
Change,
%
Specific area, m2/kg
Change,
%
Before
activation
After
activation
Before
activation
After
activation
1
Crouan
11
9
–16
910
1003
10
2
Perlite
11
14
21
838
774
–7
3
Obsidian
14
17
23
785
642
–18 The results of grain size analysis for crouan after the alkaline activation prove that
the average particle size is reduced relative to the particle size before activation. It may
be caused by crouan grains subsolution resulting in the reduction of the particle size. At the same time in the alkaline activated suspensions of perlite and obsidian there
is a tendency of particle size increase in comparison with those before activation. Hence, for crouan there is an increase of the specific area of the solid phase after the
activation and a decrease of average particle size. In case of perlite and mostly obsidian, the alkaline activation has a reverse effect:
the specific area decreases and the average particle size increases (Fig. 1). The received data of the grain size analysis suggested a scheme of aluminosilicate
structure formation that occurs during the geopolimerization. This scheme includes two
simultaneous processes: the dissolution of the aluminosilicate component and the
formation of the alkali aluminosilicate gel, which is the chemical interaction of alkali
aluminosilicate gel with unreacted grains. The chemical interaction causes the forma-
tion of a «gel layer – unreacted grain» in investigated system. Geopolymerization and Structure Formation 333 Fig. 1. The structure formation mechanism in the system “gel layer – unreacted grain of
aluminosilicate component” during the alkali activation. Fig. 1. Vivian F-I, Pradoto R GK, Moini M, Kozhukhova M, Potapov V, Sobolev K (2017) The effect of
SiO2 nanoparticles derived from hydrothermal solutions on the performance of portland
cement based materials. Front Struct Civil Eng 11(4):436–445 Sobolev K (2016) Modern developments related to nanotechnology and nanoengineering of
concrete. Front Struct Civil Eng 10(2):131–141 Wang YS, Provis JL, Dai JG (2018) Role of soluble aluminum species in the activating solution
for synthesis of silico-aluminophosphate geopolymers. Cement Concrete Compos 93:186–
195 1
Introduction Measures are taken in the building complex of Russia at the state level to improve the
efficiency and the system for determining the costs of building products. The estimated
cost of construction work includes: direct costs, overhead costs and estimated profit
(The Civil Code… 1996; On approval… 2004; Taymaskhanov et al. 2012). To ensure the effectiveness of the production of innovative products the matrix
formula by Professor Kargopolov is recommended, providing transparency and accu-
racy of calculations of the costs of production, taking into account all the territorial
conditions affecting the estimated cost of production (Kuladzhi 2014; Kargopolov
2001). Matrix Instruments for Calculating Costs
of Concrete with Multicomponent Binders T. Kuladzhi1(&), S.-A. Murtazaev2,3,4, S. Aliev2, and M. Hubaev2 1 Lomonosov North (Arctic) Federal University, Arkhangelsk, Russia
kuladzhit@list.ru
2 Millionshchikov Grozny State Oil Technical University, Grozny, Russia
3 Ibragimov Complex Research Institute, RAS, Grozny, Russia
4 Academy of Sciences of the Chechen Republic, Grozny, Russia 1 Lomonosov North (Arctic) Federal University, Arkhangelsk, Russia
kuladzhit@list.ru
2 Millionshchikov Grozny State Oil Technical University, Grozny, Russia
3 Ibragimov Complex Research Institute, RAS, Grozny, Russia
4 Academy of Sciences of the Chechen Republic, Grozny, Russia 4 Academy of Sciences of the Chechen Republic, Grozny, Russia Abstract. The paper describes methodology for determining the costs of
construction products - concrete with multicomponent binders, containing the
matrix formula by Professor M.D. Kargopolov, which is recommended a
modern micro-prediction tool for the production efficiency of building materials
and products, allowing simultaneous calculations of their costs taking into
account all the volumes of material costs: cement, fittling, components of bin-
ders, etc., as well as wage costs, depreciation etc. Keywords: Concretes with multicomponent binders Solar thermal processing Micro-prediction tools for production efficiency p
g
p
p
y
Composite building materials Matrix formula by professor M.D. Kargopolov Composite building materials Matrix formula by professor M.D. Kargopolov © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 335–338, 2019.
https://doi.org/10.1007/978-3-030-22974-0_81 References Dmitrieva TV, Strokova VV, Bezrodnykh AA (2018) Influence of the genetic features of soils on
the properties of soil-concretes on their basis. Constr Mater Products 1:69–77 Galindo Izquierdo M, Querol X, Davidovits J, Antenucci D, Nugteren HW, Fernández-Pereira C
(2009) Coal fly ash-slag-based geopolymers: microstructure and metal leaching. J Hazard
Mater 166(1):561–566 Kozhukhova NI, Zhernovsky IV, Sobolev KG (2018) Effect of variations in vitreous phase of
low-calcium aluminosilicates on strength properties of geopolymer systems. Bull BSTU
named after V.G. Shukhov 3:5–12 Shekhovtsova J, Zhernovsky I, Kovtun M, Kozhukhova N, Zhernovskaya I, Kearsley PE (2018)
Estimation of fly ash reactivity for use in alkali-activated cements - a step towards sustainable
building material and waste utilization. J Cleaner Prod 178:22–33 N. Kozhukhova et al. 334 Vivian F-I, Pradoto R GK, Moini M, Kozhukhova M, Potapov V, Sobolev K (2017) The effect of
SiO2 nanoparticles derived from hydrothermal solutions on the performance of portland
cement based materials. Front Struct Civil Eng 11(4):436–445 Wang YS, Provis JL, Dai JG (2018) Role of soluble aluminum species in the activating solution
for synthesis of silico-aluminophosphate geopolymers. Cement Concrete Compos 93:186–
195 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Materials D.Sc.(Economics) Kargopolov in (Kargopolov 2001) showed that any economic sys-
tem of economic entities can be represented as a scheme of interacting objects pro-
ducing a specific product (as part of X), from which part of the output - W in the
studied economic system is used inside the system, and the other - (Y) as the final
product is taken outside this system. Therefore, to assess the magnitude of costs and T. Kuladzhi et al. 336 results of the effectiveness of each such product, the entire production process П of an
organization can be represented by a general structure. results of the effectiveness of each such product, the entire production process П of an
organization can be represented by a general structure. Matrix formula by Professor M.D. Kargopolov is given by: P ¼
E AT
ð
Þ 1 DT C
ð1Þ ð1Þ where: P = ||pj||; j = 1; n – desired column vector of production (full) cost per unit of
production (works, services); where: P = ||pj||; j = 1; n – desired column vector of production (full) cost per unit of
production (works, services); p
(
,
);
E – single matrix n n; p
E – single matrix n n; A = ||aij||, i = 1; n, j = 1; n – matrix n n of consumption rates of own production
resources; D = ||dij||, i 2 L U R, j = 1; n – matrix of consumption rates of the primary
sources (L – variable, R – constant), T – transposition mark for matrices A and D. T – transposition mark for matrices A and D. C = ||ci||, i 2 L U R – column vector of the wholesale and procurement prices of
primary resources, and if resources are represented by value indicators in matrix D,
then in matrix C these resources, respectively, should be denoted by number – 1 (one). 3
Results and Discussion To calculate the total cost of production of reinforced concrete floor slabs using
helioforms, we used actual data on labor costs and costs of materials for the manu-
facture of reinforced concrete floor slabs of type 10-60.12 under the conditions of
Argun reinforced concrete products and structures (Table 1). Thus, Table 1 presents matrix D in such a way that it consistently (from simple to
complex products – a reinforced concrete slab) reveals indicators of the consumption of
materials for: – production of process water: 1 column; – steam production: 2 column; – production of complex binders (dry mix: cement, filler and additives “Bio-NM),
made in the scientific laboratory of building department of the Grozny State Oil
Technical University named after Academician M.D. Millionshchikov (Kargopolov
2001): 3, 4 columns Technical University named after Academician M.D. Millionshchikov (Kargopolov
2001): 3, 4 columns – production of components of concrete mixes (dry mix: complex binders - CB and
crushing residue): 5–6 columns;
3 – production of 1 m3 of concrete products under solar thermal treatment: 7–8
columns. Matrix Instruments for Calculating Costs of Concrete 337 Table 1. Matrix D of consumption rates of primary resources, incl. 3
Results and Discussion purchased, for the
production of reinforced concrete products and matrix C of wholesale procurement prices of
primary resources
Costs
Water
Steam
Binder
composition
Dry
concrete
mix
Reinforced
concrete products
with solar
thermal treatment
№
Matrix
C
KB
100
KBЗ 50
KB
100
KBЗ
50
KB 100
KBЗ 50
Price
Capital investments in
helioforms (per 1 m3 of
reinforced concrete
products), thousand
rubles/m3
0
0
0
0
0
0
0,012
0,012
1 0,012
Cement, thousand
rubles/t
0
0
0,5
0,254
0
0
0
0
2
5
Crushing residue,
thousand rubles/t
0
0
0
0 1,5
1,524
0
0 –
0,25
Filler, thousand rubles/t
0
0
0
0,254
0
0
0
0
2
1,5
BIO-NM additive,
thousand rubles/t
0
0 0,01 0,00508
0
0
0
0
4
22
Fitting, thousand
rubles/t
0
0
0
0
0
0
0,065
0,065
5
5
Electricity and fuel
0,0124 0,0414
0
0
0
0
0,2794
0,2794
6
1
Water, thousand
rubles/t
0,02365
0
0
0
0
0
0
0
7
1
Wages, thousand
rubles
0,01
0,025
0
0
0
0
0,3439
0,3439
8
1
Equipment
maintenance costs
(127.8% of salary),
thousand rubles
0,0128
0,032
0
0
0
0
0,4395
0,4395
9
1
Shop expenses (25% of
salary), thousand
rubles
0,0025 0,0064
0
0
0
0
0,086
0,086 10
1
* Deduction for social
insurance (34% for
2011 *), thousand
rubles
0,0034 0,0085
0
0
0
0
0,1169
0,1169 11
1
Plant costs (20%),
thousand rubles
0,002
0,005
0
0
0
0 0,06878 0,06878 12
1
Other, thousand rubles
0,0137
0,05
0
0
0
0
0
0 13
1 Table 1. Matrix D of consumption rates of primary resources, incl. purchased, for the
production of reinforced concrete products and matrix C of wholesale procurement prices of
primary resources 4
Conclusions Considering that the balance equation of the Nobel Prize winner in economics V.V. Leontiev is a macro-prediction tool for output of products at the national and world
levels, the matrix formula by Professor M.D. Kargopolov should be considered as an Considering that the balance equation of the Nobel Prize winner in economics V.V. Leontiev is a macro-prediction tool for output of products at the national and world
levels, the matrix formula by Professor M.D. Kargopolov should be considered as an T. Kuladzhi et al. 338 instrument for micro-prediction of cost indicators of products of any economic entities -
companies, households and other subjects, including products of cluster subjects. instrument for micro-prediction of cost indicators of products of any economic entities -
companies, households and other subjects, including products of cluster subjects. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 339–343, 2019.
https://doi.org/10.1007/978-3-030-22974-0_82 1 Standard cement chemistry notation is used.
As per this simplified notation: C = CaO, A = Al2O3, F = Fe2O3, S = SiO2, 6İ = SO3 and
T = TiO2. Characterisation of Perovskites in a Calcium
Sulfo Aluminate Cement G. Le Saout1(&), R. Idir2, and J.-C. Roux1 1 C2MA, IMT Mines Ales, Univ Montpellier, Ales, France
gwenn.le-saout@mines-ales.fr
2 CEREMA, DIM Project Team, Provins, France Abstract. Calcium sulfo aluminate cement (CSA) is a promising low CO2
footprint alternative to Portland cement. The phase assemblage of a commercial
CSA cement was investigated by a combination of XRD, SEM-EDX and
selective extraction techniques. This study focused on the composition of per-
ovsite phases present in the cement. Keywords: Calcium sulfo aluminate cement Perovskite X-ray diffraction
Scanning electron microscope References Kargopolov MD (2001) Interoperable balances of costs and results of production: theory and
practice. Monograph. Arkhangelsk: Publishing House of AGSTU, 182 p Kuladzhi TV (2014) Methodology for evaluating the effectiveness of design solutions in the
construction complex. North (Arctic) Federal University - Arkhangelsk: NAFU Publishing
House, 296 p On approval and implementation of the Methodology for determining the cost of construction
products in the territory of the Russian Federation (together with “MDS 81-35.2004 …”). Resolution of the State Construction Committee of Russia of 05.03.2004 No. 15/1 (as
amended on 06.16.2014) Taymaskhanov KE, Bataev DK-S, Murtazaev S-AY, Saidumov MS (2012) Justification of the
economic efficiency of the production of concrete composites based on technogenic raw
materials. Questions Econ Law 2:124–128 The Civil Code of the Russian Federation (part two) dated January 26, 1996 No. 14-FZ (as
amended on July 29, 2017) (as amended and added, entered into force on December 30, 2018) Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches The CSA cement was from a commercial supplier and the chemical characteristics of
the cement are given in Table 1. Table 1. Mineralogical and chemical compositions of the CSA. Mineralogical composi
determined by XRD/Rietveld analysis. Chemical analysis by X-ray fluorescence (DIN 5100
Minerals
Mass % Oxides Mass %
Anhydrite
18.3
SiO2
8.42
Gypse
2.9
Al2O3
19.1
Ye’elimite 31.4
Fe2O3
6.94
Belite
21.2
TiO2
0.76
Perovskite 11.3
K2O
0.08
Ferrite
5.4
Na2O
0.02
Merwinite
1.9
CaO
44.9
Calcite
3.5
MgO
1.27
Magnesite
1.5
SO3
15.2
aLoss
on
ignition
measured
by
calcination until 1025 °C according to
ISO 12677 Table 1. Mineralogical and chemical compositions of the CSA. Mineralogical composition
determined by XRD/Rietveld analysis. Chemical analysis by X-ray fluorescence (DIN 51001) Table 1. Mineralogical and chemical compositions of the CSA. Mineralogical composition
determined by XRD/Rietveld analysis. Chemical analysis by X-ray fluorescence (DIN 51001) Minerals
Mass % Oxides Mass %
Anhydrite
18.3
SiO2
8.42
Gypse
2.9
Al2O3
19.1
Ye’elimite 31.4
Fe2O3
6.94
Belite
21.2
TiO2
0.76
Perovskite 11.3
K2O
0.08
Ferrite
5.4
Na2O
0.02
Merwinite
1.9
CaO
44.9
Calcite
3.5
MgO
1.27
Magnesite
1.5
SO3
15.2
aLoss
on
ignition
measured
by
calcination until 1025 °C according to
ISO 12677 In order to analyze mineralogical composition, X-ray diffraction was performed on
cement with a diffractometer BRUKER D8 Advance. Powder samples were analyzed
using an incident beam angle (Cu Ka, k = 1.54 Å) varying between 5 and 70°. Software X’Pert High Score was used to process diffraction patterns and crystals were
identified using the Powder Diffraction File database. Rietveld analysis allowed
obtaining mass fractions of crystalline phases in the cement. For the microscopical investigations, powder samples were impregnated using a
low viscosity epoxy and polished down to 0.25 lm using diamond pastes. The samples
were further coated with carbon (*15 nm) and examined using a Quanta 200 FEG
scanning electron microscope (SEM) from FEI coupled to an Oxford Xmax N 80 mm2
energy dispersive X-ray spectroscopy (EDX) analyser. In order to improve the characterization of the cement, two different selective
dissolution methods were used. In the first method, the silicate phases were removed in
a solution of acid salicylic in methanol (Hjorth and Lauren 1971). In addition, a second
selective dissolution method was used to get mainly perovskite phases in CSA by
removing ye’elimite, anhydrite, gypsum with 5% Na2CO3 solution (Wang 2010). 1
Introduction Calcium sulfo aluminate cements (CSA)1 were developed by the China Building
Material Academy in the seventies. CSA have many specific properties compared to
Portland cement as fast setting, rapid hardening, shrinkage reduction. This special
cement used alone or in combination with calcium sulphates and Portland cement has
found applications such as airport runways and roads patching, selfleveling mortars, tile
adhesives grouts… (Zhang et al. 1999). This is also a promising low CO2 footprint
alternative to Portland cement due to the difference in the amount of energy used to
produce CSA cements (lower kiln temperatures and energy at the mill to grind). The
main raw materials used for making CSA cements are bauxite, limestone, clay, and
gypsum and this leads to a mineralogical composition very different than Portland
cement. While many studies have been carried out on the characterization of Portland
cement, few are available concerning CSA. In this study, we report a characterization
of a commercial CSA using Rietveld quantitative analysis and scanning electron
microscopy. 340 G. Le Saout et al. 2
Methods and Approaches The
method was modified to prevent precipitation of CaCO3 by washing the filtered sus-
pension with 6% acetic acid. The filter paper and contents were placed in an oven at
105 °C until a constant weight was reached. 341 Characterisation of Perovskites in a Calcium Sulfo Aluminate Cement 3
Results and Discussion The main phases observed in the experimental diffraction pattern (Fig. 1a) are the
orthorhombic ye’elimite C4A3S with small amount of the pseudo cubic form, belite b
and a’H- C2S and perovskites from the CSA clinker. Anhydrite II CS is also present as
mineral addition. Perovskite family has crystal structures related to the mineral per-
ovskite CT. Ferrite phase Ca2(AlxFe1-x)2O5 is usually present in CSA and its structure
is derived from that of perovskite by the substitution of Al and Fe for Ti, together with
ordered omission of oxygen atoms, which causes onehalf of the sheets of octahedral in
perovskite to be replaced by chains of tetrahedral (Taylor 1997). To obtain a good
Rietveld refinement, it is also necessary to add a perovskite phase CT (Alvarez- Pinazo
et al. 2012). The titanium dioxide is present by the use of bauxite, which usually
contains some TiO2, as raw materials in the manufacturing process of CSA clinker. The peaks associated with this cubic phase are confirmed in the XRD pattern of the
CSA after the extraction of the main phases (Fig. 1b). Fig. 1. Diffraction pattern of the cement as received (a), after extraction (b) Fig. 1. Diffraction pattern of the cement as received (a), after extraction (b) However, with the assumed stoichiometry of perovskite CT, the elemental oxide
composition TiO2 calculated from phase content deduced by Rietveld analysis is
strongly overestimated in comparison with XRF analysis. However, with the assumed stoichiometry of perovskite CT, the elemental oxide
composition TiO2 calculated from phase content deduced by Rietveld analysis is
strongly overestimated in comparison with XRF analysis. EDX analysis on polished section of CSA (Fig. 2a) and CSA after extraction
revealed an average composition of ferrite Ca1.99Al0.39Si0.10Fe1.35Ti0.08Mg0.07O5.00 not
far from the brownmillerite series Ca2(Fe2-xAlx)O5. However, the average composition
of perovskite Ca2.00Al0.30Si0.22Fe1.06Ti0.40Mg0.05O5.31 is very different to the Ca2Ti2O4
composition. These
compositions
were
similar
to
those
observed
in
the
ferrite/perovskite phases of a calcium aluminate cement (Gloter et al. 2000). We also
observed in some grains perovskite lamellae with high amount of Ti on the scale of few
micrometers (Fig. 2b). 342 G. Le Saout et al. Fig. 2. Back scattering image of the cement (a), example of EDX analysis on perovskite (b) Fig. 2. Back scattering image of the cement (a), example of EDX analysis on perovskite (b) 4
Conclusions The phase assemblage of a commercial calcium sulfo aluminate cement has been
investigated with a special attention to the ferrite- perovskite phases. The ferrite
composition is closed to the brownmillerite Ca2(Fe2-xAlx)O5 whereas the perovskite
shows heterogeneity with important substitution of Ti by Fe, Al and Si. Acknowledgements. The authors acknowledge A. Diaz (C2MA, IMT Mines Ales) for sample
preparation for SEM experiments preparation for SEM experiments Acknowledgements. The authors acknowledge A. Diaz (C2MA, IMT Mines Ales) for sample
preparation for SEM experiments 1
Introduction Nowadays many scientists are developing construction composites of new generations
(Delgado et al. 2015; Ahadi 2011). Basing on the theoretical background of geonics
(geomimetics) it is possible to reduce power consuming in construction materials
production by application of energy of geological and cosmochemical processes,
energy efficient raw material, specially treated with geological and cosmochemical
processes. So, production technologies for a wide range of composite binding materials (in-
cluding water resistant and freeze proof gypsum binders) with the application of new
raw material (Lesovik et al. 2014; Zagorodnyuk et al. 2018) with high free internal
power have been suggested. Clay rocks with an incomplete stage of mineral formation
and a sediment genesis zone, effusive rocks with an amorphous and cryptocrystalline
structure, quartz rocks of greenschist coal ranging with crystalline defects and inclu-
sions of mineral formation, gas and air inclusions, some kinds of anthropogenic
materials and others refer to these materials (Elistratkin and Kozhukhova 2018). Geonics (Geomimetics) as a Theoretical Basis
for New Generation Compositing V. Lesovik1(&), A. Volodchenko1, E. Glagolev1, I. Lashina1,
and H.-B. Fischer2 1 Belgorod State Technological University Named After V.G. Shukhov,
Belgorod, Russia
naukavs@mail.ru
2 Bauhaus-Universität Weimar, Weimar, Germany 1 Belgorod State Technological University Named After V.G. Shukhov,
Belgorod, Russia
naukavs@mail.ru
2 Bauhaus-Universität Weimar, Weimar, Germany Abstract. The article introduces basic principles of a new transdisciplinary
research area geonics (geomimetics) in the construction material science. This
research area differs from bionics, which uses knowledge about nature to solve
engineering problems. The purpose of geonics (geomimetics) is to solve engi-
neering problems with the account of knowledge about geologic and cosmo-
chemical processes. Keywords: Geonics Anthropogenic methasomatism in the construction
material science The law of consanguinity Construction composites © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 344–347, 2019.
https://doi.org/10.1007/978-3-030-22974-0_83 References Alvarez- Pinazo G, Cuesta A, Garcia-Maté M, Santacruz I, Losilla ER, De la Torre AG, Leon-
Reina L, Aranda MAG (2012) Rietveld quantitative phase analysis of Yeelimitecontaining
cements. Cem Concr Res 42:960–971 Gloter A, Ingrin J, Bouchet D, Scrivener K, Colliex C (2000) TEM evidence of perovskite-
brownmillerite coexistence in the Ca(AlxFe1-x)O2.5 system with minor amounts of titanium
and silicon. Phys. Chem. Miner 27:504–513 y
Hjorth L, Lauren K-G (1971) Belite in Portland cement. Cem Concr Res 1:27–40 Taylor HFW (1997) Cement Chemistry. Thomas Telford, London Wang J (2010) Hydration mechanism of cements based on low-CO2 clinkers containing belite,
ye’elimite and calcium alumino ferrite. PhD dissertation, University of Lille I, France Zhang L, Su M, Wang Y (1999) Development of the use of sulfo- and ferroaluminate cements in
China. Adv Cem Res 11:15–21 343 Characterisation of Perovskites in a Calcium Sulfo Aluminate Cement Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches The law of consanguinity has been formulated within the theoretical backgrounds of
geonics (geomimetics). It implies designing layered composites and maintenance
systems at nano-, micro-, and macro- levels similar to basic matrix that enhances Geonics (Geomimetics) as a Theoretical Basis 345 significantly material adhesion and durability. The implementation of this law allows
creating a composite which components have close deformation and temperature
characteristics. The theory of anthropogenic methasomatosis in the construction material science
has been suggested. This stage of composite materials evolution is characterized by
composite ability to adjust to changing conditions during building and construction
service. Designing construction composites with the account of anthropogenic
methasomatosis theory in the construction material science provides possibility of self-
healing defects, which appear during construction and building service and design, they
are so-called “smart” composites. 4
Conclusions Hence, monodisciplinary and interdisciplinary approaches in the construction material
science promoted developing a wide range of construction composites which main task
is construction of solid and durable structures. Design and creation of materials for
environment optimization “a man-material-environment” is a complex problem, which
requires united work of scientists of different fields. A single way to solve this problem
is transdisciplinary approach as a way of widening the scientific world view, which
requires considering phenomena beyond a single science. Acknowledgements. The study is carried out in the framework of the State Task of the RF
Ministry of Education and Science No. 7.872.2017/4.6. Development of principles for the design
of ecologically positive composite materials with prolonged bioresistance 2017–2019. V. Lesovik et al. These materials are designed with the account of the system interacting with the
environment. This system allows reacting to the external actions by defects “self-
healing” and having positive impact on the triad “a man-material-environment”. This approach has been tested on the composite bindings based on calc-sinter,
which create favorable conditions for at the early stages of structure formation and
system hardening. It decreases stress in the hardening composite and as a consequence
decreases the amount and the size of micro-cracks that predetermines technical and
economic efficiency of the composite binding based on tuff, especially in hot climate. Volcanic tuff is known to be a heteroporous rock. Pore space of this rock is rather
complex in form and combines pores of different sizes. Water in this rock is in complex interaction with its mineral grid, whose boundaries
and ratio are relative and change constantly: vapour; chemically and physically
bounded water; free or gravitational water. In a hot climate, with a deficit of liquid phase in the concrete itself, tuff particles in
the binding mixture composition, during the hardening process will release the capil-
lary water, it will activate structure formation processes and synthesize more massive
structure of the materials during concrete hardening and utilization. The micro-cracks appearing during service at different conditions will self-heal by
interaction of water in tuff particles with unreacted cement minerals. During service tuff
particles release saved capillary water and that will result in structure formation acti-
vation and synthesizing more massive materials structure during concrete hardening
and operation. These are so-called smart composites. Ahadi P (2011) Applications of nanomaterials in construction with an approach to energy issue.
Adv Mater Res 261–263:509–514 3
Results and Discussion Principles for productivity enhancement of wall materials manufacturing using sandy
and clay rocks with an incomplete stage of clay formation and industrial wastes in
hydrothermal processing at atmospheric pressure have been suggested. It has been
found that aeolian-sedentary-diluvia clay rocks of the Quaternary period are the
products of the initial clay formation stage including metastable atelene minerals of
nano-level and non-rounded finely dispersed quartz and are suitable as a raw material
for autoclaved silicate materials production. Application of sandy and clay rocks instead of traditional quartz sand in silicate
materials production enhances raw mixture formation, hardens raw bricks by 4–11
times, that allows developing the production technique of highly-hollowed construction
products. Application of these rocks allows widening the range of autoclaved raw material
base, decrease energy-output ratio of their production, improve the ecological state of
the environment and create comfortable conditions for human life. Within the law of consanguinity it has been found that rocks independent of their
genesis (magmatic, methamorphic and aqueous) with banded texture, whose layers are
introduced by minerals whose stress related characteristics and thermal-expansion
coefficient differ significantly, are non-durable and have anisotropy coefficient 7–9
higher comparing with prototypes with anisotropy coefficient 2–3. Application of the
suggested mixtures increases brickwork breaking limit by 3–5 times. It is explained by
the contact zone microstructure, for example, ceramic brick and binding matrix. The
designed binding matrix and wall material are nearly sole block sample and con-
structions with a traditional binding matrix have a distinct contact zone – the weakest
place of the samples. This law allows creating restoration mixtures, plastering mate-
rials, brickwork and restoration compositions of new generations for every walling. With the account of geonics theoretical background a wide range of acoustic,
insulating, and construction insulating composites based on foam glass have been
suggested. Designing construction composites with the account of anthropogenic methaso-
matosis theory in the construction material science provides possibility of defects self-
healing which appear during construction and building service and design so-called
“smart” composites. 346 Ahadi P (2011) Applications of nanomaterials in construction with an approach to energy issue.
Adv Mater Res 261–263:509–514
Delgado JMPQ, Cerný R, de Lima AGB, Guimarães AS (2015) Advances in building
technologies and construction materials. Adv Mater Sci Eng 2015:1–3 no. 312613 Delgado JMPQ, Cerný R, de Lima AGB, Guimarães AS (2015) Advances in building
technologies and construction materials. Adv Mater Sci Eng 2015:1–3 no. 312613 Elistratkin MY, Kozhukhova MI (2018) Analysis of the factors of increasing the strength of the
non-autoclave aerated concrete. Constr Mater Prod 1(1):59–68 References Ahadi P (2011) Applications of nanomaterials in construction with an approach to energy issue. Adv Mater Res 261–263:509–514 Delgado JMPQ, Cerný R, de Lima AGB, Guimarães AS (2015) Advances in building
technologies and construction materials. Adv Mater Sci Eng 2015:1–3 no. 312613 Geonics (Geomimetics) as a Theoretical Basis 347 Elistratkin MY, Kozhukhova MI (2018) Analysis of the factors of increasing the strength of the
non-autoclave aerated concrete. Constr Mater Prod 1(1):59–68 Lesovik VS, Chulkova IL, Zagorodnjuk LH, Volodchenko AA, Popov DY (2014) The role of the
law of affinity structures in the construction material science by performance of the restoration
works. Res J Appl Sci 9(12):1100–1105 pp
Zagorodnyuk L, Lesovik VS, Sumskoy DA (2018) Thermal insulation solutions of the reduced
density. Constr Mater Prod 1(1):40–50 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Regularities in the Formation of the Structure
and Properties of Coatings Based on Silicate
Paint Sol V. Loganina(&), E. Mazhitov, and V. Demyanova Department “Quality Management and Technology of Construction Production”,
Penza State University of Architecture and Construction, Penza, Russia
loganin@mail.ru Department “Quality Management and Technology of Construction Production”,
Penza State University of Architecture and Construction, Penza, Russia
loganin@mail.ru Abstract. The authors established a higher quality of the appearance of coat-
ings based on silicate paint sol in comparison with coatings based on silicate
paints. Information is provided on the regularities in the formation of the quality
of the appearance of coatings on the basis of silicate paint sol is that the silicate
paint sol has a higher value coefficient of the wetting and spreading on the
cement substrate in comparison with silicate paint. It was established that when
using a paint brush on the cement substrate, there has been some slowing of the
rise time of the structure of silicate paint compositions based on polysilicate
solutions. A higher value of the adhesion work based on polysilicate solutions
were indentified, indicating the strong force of the paint and the cement
substrate. Keywords: Sol silicate paint Polysilicate binder Coatings Wetting
Coating quality Keywords: Sol silicate paint Polysilicate binder Coatings Wetting
Coating quality 1
Introduction The problem of reliability and durability of protective decorative coatings of exterior
walls of buildings is one of the topical scientific and technical objectives in materials
science. It is known that the longevity of coatings depend on the type of binder, the
technology of applying the paint composition, operating conditions, etc. (Ailer 1982). In the practice of finishing works, silicate paints, which are a suspension of pigments
and fillers in liquid glass of potassium, proved to be very useful. To improve the
performance of coatings based on silicate paints, it is important to develop methods for
modifying liquid glass. Analysis of patent and scientific-technical literature shows that
one way of modification is the introduction of organosilicon and other polymeric
compounds into the binder. In works (Figovsky et al. 2012) there is an increased
durability of silicate coatings when imposing of polymeric compounds. It is of interest
to use polysilicates in silicate paints as film-forming substances that provide higher
performance properties of coatings. However, at present time the questions of the
formation of the structure and properties of coatings based on sol silicate paints have
not been studied, the questions of mechanism for improving the operational properties
of coatings based on them have not been considered. Polysilicates are characterized by © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 348–351, 2019. https://doi.org/10.1007/978-3-030-22974-0_84 Regularities in the Formation of the Structure and Properties 349 a broad range of the degree of polymerization of anions and are dispersions of colloidal
silica in an aqueous solution of alkali metal silicates. We have established a paint
composition based on a polysilicate binder obtained by mixing liquid glass with a silica
sol (Loganina et al. 2018a, b). It was found, that coatings based on polysilicate solu-
tions are characterized by faster curing. Films based on polysilicate solutions have
higher cohesive strength. The tensile strength of a film based on potassium liquid glass
is Rp = 0.392 MPa, and the tensile strength of a film based on a polysilicate solution
(15% Nanosil 20) is 1.1345 MPa. The paint forms a coating characterized by a uniform
homogeneous surface. To study the regularities in the formation of the quality of the appearance of
coatings on the based on sol silicate paint, the character of filling on a porous cement
substrate was considered. 2
Methods and Approaches The character of the filling of the sol of the silicate paint was evaluated. The method of
determining filling consisted in applying five parallel strips of paint and determining
the degree of spreadability according to the number of stuck bands. Paint with an
operating viscosity was applied to a glass plate measuring 200 100 1.2 mm. The
spreading of five parallel strips was evaluated on a ten-point scale of filling. The surface tension of the paint was determined by the drop method (stalagmo-
metric method). Work of adhesion of the paint to a cement substrate was calculated
using the Dupre - Young thermodynamic equation. The wetting operation and
spreading coefficient were determined. The quality of the appearance of the coatings was estimated from the surface
roughness Ra by the method of scanning probe microscopy (SPM) (Chizhik and
Syroezhkin 2010). 3
Results and Discussion Previously, the rheological type of the solutions was determined. The rheological
properties were evaluated by the indicators of conventional viscosity according to BЗ-
4, critical shear stress with instrument Reotest-2. It is found, that all systems are typical
pseudoplastic bodies. In the region of slow flow, the viscosity of polysilicate solutions
gradually declined with increasing shear stress. Analysis of data (Table 1) shows, that silicate paints based on polysilicate solutions
have a long filling time. Thus, the time for bottling for a paint based on liquid glass is
6 min, and for a paint based on a polysilicate solution - 8 min 40 s. The degree of filling
is satisfactory (no more than 10 min). For paints based on a polysilicate solution, a large
work of adhesion to the substrate is characteristic. So, work of the adhesion of paint
based on the potassium polysilicate solution to the substrate is 108.17 mN/m, while
work of the adhesion of the paint based on potassium liquid glass is 96.82 mN/m. A higher value of work of the adhesion of paint based on polysilicate solutions indicates
a stronger interaction of the paint and the cement substrate. V. Loganina et al. V. Loganina et al. 350 Table 1. Test results
The name of indicators
Name of the paint composition
Based on potassium
liquid glass
Based on potassium
polysilicate solution
Surface roughness, Ra, [lm]
16,208
10,880
The contact angle of wetting
50,9
51,6
Surface tension of the paint
composition, [mN/m]
59,38
66,73
Filling the colorful composition *
7 min 40 s
9
8 min 40 s
9
Adhesion work, [mN/m]
96,82
108,17
Wetting operation, [mN/m]
37,44
41,44
Cohesion work, [mN/m]
118,76
133.46
Coefficient of spreading [mN/m]
−21,94
−25,29
Coefficient of wetting
0,815
0,81
Note: * Above the line are the values of time of restoration of the paint structure, below the line -
the value of filling Table 1. Test results Name of the paint composition Note: * Above the line are the values of time of restoration of the paint structure, below the line -
the value of filling The work of wetting paints based on polysilicate solution is higher, which indicates
better wetting of the paint surface of the cement substrate. Thus, the work of wetting
the sol of silicate paint on the basis of potassium polysilicate solution is 41, 44 mN/m,
and on the basis of potassium liquid glass - 37.44 mN/m. Regularities in the Formation of the Structure and Properties Regularities in the Formation of the Structure and Properties Higher waterproof properties of coatings based on sol silicate paint are caused, in
our
opinion,
by
the
structure
of
the
coating. Scanning
probe
microscopy
(SPM) methods were used to estimate the local structure of the coating surface. It is
established, that the surface of coatings based on potassium liquid glass contains a
certain number of pores of the nanometric range, differing in size and shape. The
maximum pore size is 19.8 lm. Pores with a diameter of 18.85 to 19.6 lm are mainly
present, whereas in the coating based on the potassium polysilicate solution there are
two groups of pores: from 19.25 to 19.8 lm and from 20.0 to 20.6 lm. The value of
the maximum pore size is 21.2 lm. In the coating based on the polysilicate solution is
observed a more uniform pore size distribution. Coatings based on the developed paint are characterized by high adhesion (1.1–
1.3 MPa), coefficient of vapor permeability - 0,00878 mg/m * hPa. 4
Conclusions The properties of the paint and coating based on it meet the requirements for coatings for
exterior decoration of buildings, have higher adhesion, sufficient vapor permeability. 3
Results and Discussion When the sol of the silicate
paint is applied to the substrate, the wetting and spreading coefficient increases, which
indicates more favorable conditions for the formation of the quality of the appearance. The surface roughness of the coating based on silicate paint is Ra = 16.208 lm, and
based on the potassium polysilicate solution, Ra = 10.880 lm. The quality of the
appearance of the surface of the coatings formed by the sol with silicate paint, in
accordance with GOST 9.032-74 ** “Unified system for protection against corrosion
and aging. Coatings for paint and varnish, Groups, technical requirements and desig-
nations” is graded IV class, and on the basis of liquid glass - V class. g
q
g
Testing of solution samples, colored with sol by silicate paint, was carried out for
frost resistance by alternating thawing and freezing. Appearance of coatings was
assessed according to GOST 6992-68. “Coatings for paint and varnish. Test method for
resistance to atmospheric conditions”. It was found, that coatings on the basis of the
developed composition had withstood 40 test cycles, while the coating condition after
40 test cycles was estimated at I.1 points, which corresponds to the coating condition
with no color change, chalking, mud retention. To assess the waterproof properties of coatings, tests were carried out of solution
samples stained with silicate and sol silicate paints. After curing of the coatings, water
absorption was determined upon capillary suction. It was found, that water absorption
by capillary suction of samples stained with silicate paint is 4.4%, and stained with
silicate paint - 4.6%. The lower value of water saturation of samples colored with sol by
silicate paint indicate a change in the pore size in the coating structure as compared to
the coating based on silicate paint. 351 (
)
y
(
g
)
p
Chizhik SA, Syroezhkin SV (2010) Methods of scanning probe microscopy in micro- and
nanomechanics. Instr Meas Methods 1:85–94 Figovsky O, Borisov Yu, Beilin D (2012) Nanostructured binder for acid-resisting builder
materials. J Sci Israel-Technol Advant 14(1):7–12 References Ailer P (1982) The chemistry of silica (Transl. from English), Part 1. Mir, Moscow, p. 416 Ailer P (1982) The chemistry of silica (Transl. from English), Part 1. Mir, Moscow, p. 416 Chizhik SA, Syroezhkin SV (2010) Methods of scanning probe microscopy in micro- and
nanomechanics. Instr Meas Methods 1:85–94 Figovsky O, Borisov Yu, Beilin D (2012) Nanostructured binder for acid-resisting builder
materials. J Sci Israel-Technol Advant 14(1):7–12 Loganina VI, Kislitsyna SN, Mazhitov YB (2018a) Development of sol-silicate composition for
decoration of building walls. Case Stud Constr Mater 9:e00173 Loganina VI, Kislitsyna SN, Mazhitov YB (2018a) Development of sol-silicate composition for
decoration of building walls. Case Stud Constr Mater 9:e00173 Loganina VI, Kislitsyna SN, Mazhitov YB (2018b) Properties of polysiylate binders for sol-
silicate pains. Adv Mater Res 1147:1–4 Loganina VI, Kislitsyna SN, Mazhitov YB (2018b) Properties of polysiylate binders for sol-
silicate pains. Adv Mater Res 1147:1–4 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction Modern rotary furnaces of cement industry are characterized by accumulation and
circulation of alkali salts in the furnace system (Luginina 2002). As a result, the R2O
content of furnace charge may reach 3.5% before the sintering zone, and in some plants
this value may even reach 10%. The formation of calcium aluminate ferrite is not
observed in furnace charges of alumina industry characterized by high Na2O content
due to the formation of aluminates and sodium ferrites (Lisiyenko 2004). Hence, as
may be expected, the accumulation and circulation of alkali salts will lead to the
situation that occurs in furnaces of alumina industry. Raw mixes from analytical reagents used to obtain C4AF are characterized by the
formation of sodium compounds instead of calcium compounds (Kovalyov 2015). It
will allow obtaining a clinker of lighter shade. Thus, the purpose of this study is to determine the possibility of adjusting the
brightness coefficient of portland cement by introducing sodium oxide into the raw
mix. Influence of Sodium Oxide on Brightness
Coefficient of Portland Cement Clinker D. Mishin(&) and S. Kovalyov D. Mishin(&) and S. Kovalyov Department of Technology of Cement and Composite Materials,
Chemical Technology Institute, Belgorod State Technological University
named after V.G. Shukhov, Belgorod, Russia
mishinda.xtsm@yandex.ru Department of Technology of Cement and Composite Materials,
Chemical Technology Institute, Belgorod State Technological University
named after V.G. Shukhov, Belgorod, Russia
mishinda.xtsm@yandex.ru Abstract. The paper is devoted to the possibility of adjusting the reflection
factor of portland cement clinker. For this purpose, Na2CO3 is introduced into
the slurry of CJSC Belgorod Cement Plant. The influence of Na2O on brightness
coefficient of the crushed clinker is established at a burning temperature of
1250–1300 °C. With the increase of Na2O concentration up to 1–2% the
brightness coefficient is reduced and the increase of Na2O in the range of
3.5–7% leads to sharp increase of the reflection factor and CaOfree content. Keywords: Sodium oxide Brightness coefficient White cement
Free calcium oxide Keywords: Sodium oxide Brightness coefficient White cement
Free calcium oxide Keywords: Sodium oxide Brightness coefficient White cement © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 352–355, 2019.
https://doi.org/10.1007/978-3-030-22974-0_85 The dried slurry of CJSC Belgorod Cement Plant with the following modular char-
acteristics: KH = 0.91; n = 2.23; p = 1.29 was used as a raw mix (Table 1). Table 1. Chemical composition of slurry of CJSC Belgorod Cement Plant, % Sodium carbonate of CH classification was introduced into the raw mix in the
amount of 0.5; 1; 2; 3.5; 5; 7% of Na2O in equivalent of ignited basis. Tablets weighing
2 g were formed from obtained mix under manual pressure. In order to avoid Na2O
volatilization during roasting, the samples were covered with a platinum cup. Roasting
of samples was carried out in a laboratory furnace with isothermal time of 20 min. The
heating rate of the furnace made 10 °C/min. The influence of alkali salts on the formation of aluminate-ferrite phase was esti-
mated according to the content of free calcium oxide and the brightness coefficient of
samples. The CaOfree content in a clinker was defined through ethyl-glycerate method
(Butt and Timashev 1973). The whiteness (brightness coefficient) of a clinker was
defined via FB-2 reflection meter on a reference polished barium sulfate plate. 2
Methods and Approaches The dried slurry of CJSC Belgorod Cement Plant with the following modular char-
acteristics: KH = 0.91; n = 2.23; p = 1.29 was used as a raw mix (Table 1). The dried slurry of CJSC Belgorod Cement Plant with the following modular char-
acteristics: KH = 0.91; n = 2.23; p = 1.29 was used as a raw mix (Table 1). 353 Influence of Sodium Oxide on Brightness Coefficient Table 1. Chemical composition of slurry of CJSC Belgorod Cement Plant, %
Losses on ignition SiO2
Al2O3 Fe2O3 CaO
MgO K2O Na2O SO3
Other
34.8
14.23 3.59
2.78
43.12 0.6
0.4
0.11
0.09 0.37 Table 1. Chemical composition of slurry of CJSC Belgorod Cement Plant, %
Losses on ignition SiO2
Al2O3 Fe2O3 CaO
MgO K2O Na2O SO3
Other
34.8
14.23 3.59
2.78
43.12 0.6
0.4
0.11
0.09 0.37 3
Results and Discussion The compositions burned at 1250 and 1300 °C were analyzed. The roasting temper-
ature was chosen based on the following: at given temperatures the formation of belite
was complete, and the synthesis of alite had not started yet. As a result, the formation of
silicate phases would not change the content of free calcium oxide. At a temperature of 1250 °C (Fig. 1) at the initial stage (up to 1% Na2O) the
content of free calcium oxide is reduced alongside with the brightness coefficient (BC). The brightness coefficient almost does not change with Na2O concentration in the range
of 1–3.5% and the content of free calcium oxide. With the introduction of over 3.5% of
Na2O the CaOfree content and the brightness coefficient sharply increase. 50.00
52.00
54.00
56.00
58.00
60.00
62.00
5.00
6.00
7.00
8.00
9.00
10.00
11.00
0
2
4
6
8
СаОfree content
Brightness coefficient, %
Na2O volume, %
СаОсв. КЯ
СаОfree
BC
Fig. 1. Influences of Na2O on brightness coefficient and content of free calcium oxide at 1250 °C
roasting temperature. 50.00
52.00
54.00
56.00
58.00
60.00
62.00
5.00
6.00
7.00
8.00
9.00
10.00
11.00
0
2
4
6
8
СаОfree content
Brightness coefficient, %
Na2O volume, %
СаОсв. КЯ
СаОfree
BC Fig. 1. Influences of Na2O on brightness coefficient and content of free calcium oxide at 1250 °C
roasting temperature. 354 D. Mishin and S. Kovalyov At a roasting temperature of 1300 °C (Fig. 2) a similar situation is observed. With
the amount of introduced Na2O up to 1–2% the considered characteristics are reduced. Sharp increase of reflection factor and CaOfree content was also observed in the range
of 3.5–7% of Na2O. 40.00
42.00
44.00
46.00
48.00
50.00
52.00
54.00
56.00
58.00
4.00
6.00
8.00
10.00
12.00
14.00
16.00
18.00
0
2
4
6
8
СаОfree content
Brightness coefficient, %
Na2O volume, %
СаОсв. КЯ
СаОfree
BC
Fig. 2. Influence of Na2O on reflection factor and content of free calcium oxide at 1300 °C
roasting temperature. Fig. 2. Influence of Na2O on reflection factor and content of free calcium oxide at 1300 °C
roasting temperature. The relatively low content of free calcium oxide without alite formation may be
caused by the formation of intermediate phases, for example carbonate spurrite. The
presence of Na2CO3 in the system fosters its formation. Hence, the increase of Na2O
content at the initial stage leads to CaOfree decrease. 4
Conclusions The introduction of up to 2–3.5% of Na2O leads to the decrease in the brightness
coefficient of samples due to the reduction of free calcium oxide content. Apparently,
this is caused by the formation of intermediate phases – carbonate spurrite. The Na2O content of over 3.5% increases the brightness coefficient of samples. This is caused by the formation of aluminates and sodium ferrites instead of aluminates
and calcium aluminate ferrites. Acknowledgements. The study is implemented in the framework of the Flagship University
Development Program at Belgorod State Technological University named after V.G. Shukhov,
using the equipment of High Technology Center at BSTU named after V.G. Shukhov. 3
Results and Discussion The sharp increase of free calcium oxide content starting from Na2O concentration
of approximately 3.5% confirms the formation of ferrites or sodium aluminates instead
of calcium aluminate ferrites in clinker systems, since such interaction results in the
emission of additional amounts of calcium oxide and may be presented by the fol-
lowing reactions: 3CaO Al2O3 þ Na2O ¼ 2NaAlO2 þ 3CaO
ð1Þ
4CaO Al2O3 Fe2O3 þ 2Na2O ¼ 2NaFeO2 þ 2NaAlO2 þ 4CaO
ð2Þ ð1Þ ð2Þ The change dependence of brightness coefficient within samples on Na2O content
in a clinker correlates with the change dependence of free calcium oxide content. This
may be explained by the fact that CaOfree crystals are white. Therefore, the increase or
reduction of its content leads to the corresponding change of the brightness coefficient
of a clinker. Based on above conclusions, the sharp increase of the brightness coeffi-
cient with the increase in Na2O concentration of over 3.5% may also be caused by the
complexity of aluminate-ferrite phase. This phase is the most colored part of a clinker
(Zubekhin et al. 2004). Hence, the change of its composition and quantity resulting
from the formation of aluminates and sodium ferrites will be characterized by the
corresponding change of the brightness coefficient of a clinker. 355 Influence of Sodium Oxide on Brightness Coefficient © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 356–359, 2019.
https://doi.org/10.1007/978-3-030-22974-0_86 1
Introduction Currently, white cement is one of the most demanded building materials due to its wide
building and technological properties. The demand for this cement grows with the rate
of city development. However, the development of white cement industry is limited by
strict requirements to the quality of raw materials. To produce cement of graded
whiteness, the content of iron oxide in the clinker should not exceed 0.5% (Zubekhin
et al. 2004). The number of stock deposits that can provide such content of iron oxide
in clinker is relatively low and will gradually decrease with their depletion. Thus, the development of new technologies extending the range of raw material
sources for white cement by involving components with higher iron content becomes
very urgent. References Butt YM, Timashev VV (1973) Practical Guide on Chemical Technology of Binding Materials:
Study Manual. Higher School, p 504 Zubekhin AP, Golovanova SP, Kirsanov PV (2004) White portland cement. Publishing House of
Izvestiya Vuzov Journal, Rostov-On-Don, North Caucasian Region, p 264 (Zubekhin AP
(ed)) Kovalyov SV (2015) Essentially new method of clinker bleaching with high iron content. In:
Kovalyov SV, Mishin DA (eds) International seminar-competition of young scientists and
graduate students working in the field of binding materials, concrete and dry mixes: collection
of articles: 180 pages. Alitinform Publishing House, Saint Petersburg, pp 29–37 Lisiyenko VG (ed) (2004) Rotary furnaces: thermal engineering, management and ecology. Teplotekhnik, p 688 (Reference edition. Lisiyenko VG, Shchelokov YM, Ladygichev MG)
Luginina IG (2002) Selected articles. Publishing House of BSTU named after V.G. Shukhov,
Belgorod p 302 Lisiyenko VG (ed) (2004) Rotary furnaces: thermal engineering, management and ecology. Teplotekhnik, p 688 (Reference edition. Lisiyenko VG, Shchelokov YM, Ladygichev MG)
Luginina IG (2002) Selected articles. Publishing House of BSTU named after V.G. Shukhov,
Belgorod, p 302 Lisiyenko VG (ed) (2004) Rotary furnaces: thermal engineering, management and ecology. Teplotekhnik, p 688 (Reference edition. Lisiyenko VG, Shchelokov YM, Ladygichev MG) Luginina IG (2002) Selected articles. Publishing House of BSTU named after V.G. Shukhov,
Belgorod, p 302 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Production of Bleached Cement D. Mishin(&) and S. Kovalev D. Mishin(&) and S. Kovalev Department of Technology of Cement and Composite Materials,
Institute of Chemical Technology, Belgorod State Technological University
named after V.G. Shukhov, Belgorod, Russia
mishinda.xtsm@yandex.ru Abstract. The present paper studies a new method of bleaching that allows
extending the range of raw materials for white cement. This paper investigates
the separate introduction of mineralizers into raw slurry of CJSC Belgorod
Cement plant containing 2.78% of Fe2O3. The separate introduction of miner-
alizers provides bleaching effect and complete consumption of free calcium
oxide at the temperature of 1250 °C. The bleaching is caused by the reduced
content and changed composition of ferroaluminate phase. Keywords: Clinker bleaching White cement Separate introduction
Na2CO3 CaF2 Mineralizers Keywords: Clinker bleaching White cement Separate introduction
Na2CO3 CaF2 Mineralizers Table 1. Chemical composition of slurry of CJSC Belgorod Cement plant [%]
Loss on ignition SiO2
Al2O3 Fe2O3 CaO
MgO K2O Na2O SO3
other
34.8
14.23 3.59
2.78
43.12 0.6
0.4
0.11
0.09 0.37 The effect of mineralizers on the phase composition of the clinker was established
using XRD analysis using ARL™X’TRA Powder Diffractometer (Switzerland). The completion of clinker formation process was assessed by the content of free
calcium oxide. The content of CaOfree was determined by ethyl-glycerate method
(Boutt and Timashev 1973). The whiteness grade (brightness coefficient) of the clinker was determined by FB-2
reflection meter using reference polished plate of barium sulphate. 2
Methods and Approaches Materials and Methods. The raw mixture with high content of Fe2O3 was represented
by dried slurry of CJSC Belgorod Cement plant (Table 1) with the following module
characteristics: LSF = 0.91; n = 2.23; p = 1.29. The raw mixture was doped by mineralizers Na2CO3, CaF2 and 2C2S·CaF2 calcu-
lated as ignition basis over 100%: The final mixture contained 3.5% of sodium car-
bonate as R2O, 1.5% of calcium fluoride and 8.11% of synthesized 2C2S·CaF2 (1.5% if
expressed as CaF2). The mineralizers were introduced separately (Mishin et al. 2016). 357 Production of Bleached Cement 3
Results and Discussion The implementation of separate introduction of mineralizers leads to bleaching of the
samples at the temperature of 1250–1300 °C (Fig. 1(1)). The increase of the burning
temperature from 1250 to 1300 °C causes gradual return of clinker color to typical
black (Fig. 1(1), (2) and (3)). 1)
2)
3)
Fig. 1. Appearance of clinker samples produced by separate introduction at burning temperature
of: (1) 1250 °C; (2) 1275 °C; (3) 1300 °C. Fig. 1. Appearance of clinker samples produced by separate introduction at burning temperature
of: (1) 1250 °C; (2) 1275 °C; (3) 1300 °C. In this temperature interval, complete consumption of free calcium oxide is
observed, which indicates the completion of clinker formation processes (Table 2). Table 2. Characteristics of clinker samples produced by separate introduction of 2C2S·CaF2
mineralizer Table 2. Characteristics of clinker samples produced by separate introduction of 2C2S·CaF2
mineralizer Burning temperature [°C] 1250
1250 1275 1300 1400*
Cooling method
(water) (air)
(air)
(air)
(air)
Content of CaOfree [%]
0.55
0.5
0.3
0.31
0.10
Brightness coefficient [%]
46
41
37.5
30
31
* common plant clinker 358 D. Mishin and S. Kovalev Shooting angle, 2θ, degree
Shooting angle, 2θ, degree
Fig. 2. Phase composition of clinker samples: (1) no additives at 1250 °C; produced by separate
introduction at: (2) 1250 °C; (3) 1300 °C. Shooting angle, 2θ, degree Fig. 2. Phase composition of clinker samples: (1) no additives at 1250 °C; produced by separate
introduction at: (2) 1250 °C; (3) 1300 °C. According to the phase composition analysis, at the burning temperature of 1250 °C
(Figs. 2(1), (2)), in the samples produced by separate introduction, the composition of
ferroaluminate phase C4AF (d, Å: 7.314; 2.679; 2.637) shifts into the region of com-
positions with a higher iron content and approaches that of C6AF2 (d, Å: 7.375; 2.656). The intensity of reflections of the ferroaluminate phase decreases, which tells about the
decrease in its content in the clinker. Ferroaluminate phase is the most coloring among
the clinker components (Zubekhin et al. 2004). In this connection, the decreased amount
of ferroaluminate phase leads to increased brightness coefficient of the clinker. The increased burning temperature up to 1300 °C (Fig. 2(3)) causes increased
intensity of ferroaluminate phase reflections (d, Å: 7.375; 2.659), which witnesses its
increased content in the clinker. As a result, the brightness coefficient of the clinker
reduces from 41% down to 30%. The increase in the brightness coefficient is connected with decreased content of
calcium ferroaluminate phases in the clinker: C4AF, C6AF2. The separate introduction of mineralizer allows bleaching the clinker with the iron
oxide content of more than 0.5%. With the iron oxide content in raw mixture of 2.78%,
the brightness coefficient increases from 31% up to 46%. 4
Conclusions The separate introduction of mineralizer allows bleaching the clinker with the iron
oxide content of more than 0.5%. With the iron oxide content in raw mixture of 2.78%,
the brightness coefficient increases from 31% up to 46%. The increase in the brightness coefficient is connected with decreased content of
calcium ferroaluminate phases in the clinker: C4AF, C6AF2. Production of Bleached Cement 359 The implementation of separate introduction of mineralizer will allow extending the
range of sources for white cement production involving raw components with increased
iron content. The separate introduction of mineralizer in the presence of increased iron oxide
content allows achieving complete consumption of free calcium oxide at 1250 °C. Acknowledgements. The work is realized in the framework of the Program of flagship uni-
versity development on the base of Belgorod State Technological University named after V.G. Shoukhov, using equipment of High Technology Center at BSTU named after V.G. Shoukhov. Mishin DA, Kovalev SV, Chekulaev VG (2016) Reason for reduced efficacy of mineralizers for
burning Portland cement clinker. Bulletin of BSTU n.a. V.G. Shoukhov, no 5, p 161–166 (in
Russian) Boutt YM, Timashev VV (1973) Practical guide on chemical technologies of binders: guide.
Higher school, Moscow, 504 p. (in Russian) Zubekhin AP, Golovanova SP, Kirsanov PV (2004) White Portland cement. Zubekhin AP
(ed) J. “Bulletin of HEIs North-Caucasus region”, Rostov on Don, 264 p (in Russian) Multicomponent Binders
with Off-Grade Fillers S.-A. Murtazaev1,2,3, M. Salamanova1,2(&), M. Saydumov1,
A. Alaskhanov1, and M. Khubaev1 1 Millionshchikov Grozny State Oil Technical University, Grozny, Russia
Madina_salamanova@mail.ru
2 Ibragimov Complex Research Institute, RAS, Grozny, Russia
3 Academy of Sciences of the Chechen Republic, Grozny, Russia Abstract. The paper deals with issues related to development of multicom-
ponent binders (MCB) and high-quality concretes based on them. The pro-
duction of such binders is based on the use of finely divided mineral additives of
natural and technogenic origin. Particular attention is paid to the aggregate, the
strength of coarse aggregate should be at least 20% higher than the strength of
concrete, and the maximum particle size should not exceed 8–20 mm. At pre-
sent, considerable experience was accumulated for production of multicompo-
nent binders, and the results of studies conducted in this direction showed that
the raw material potential of the Republic allowed obtaining high-quality class
B30-40 concrete, and if we expanded the geography of the use of natural
resources by regions of the North Caucasus, we could produce concretes with
higher strength. Keywords: High-quality concretes Composite binders
Reactive mineral components Volcanic ash Thermal power plant (TPP) ash
Fractionated filler Keywords: High-quality concretes Composite binders Reactive mineral components Volcanic ash Thermal power plant (TPP) ash
Fractionated filler © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 360–364, 2019.
https://doi.org/10.1007/978-3-030-22974-0_87 References Boutt YM, Timashev VV (1973) Practical guide on chemical technologies of binders: guide. Higher school, Moscow, 504 p. (in Russian) Mishin DA, Kovalev SV, Chekulaev VG (2016) Reason for reduced efficacy of mineralizers for
burning Portland cement clinker. Bulletin of BSTU n.a. V.G. Shoukhov, no 5, p 161–166 (in
Russian) Zubekhin AP, Golovanova SP, Kirsanov PV (2004) White Portland cement. Zubekhin AP
(ed) J. “Bulletin of HEIs North-Caucasus region”, Rostov on Don, 264 p (in Russian) Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Materials As part of the work carried out in this direction, we developed formulations of mul-
ticomponent binders, which include mineral additives of natural and technogenic
origin. The North Caucasus has large reserves of natural raw materials for these devel-
opments, the chemical analysis of the mineral components used in the studies is shown
in Table 1. Table 1. The chemical composition of mineral components, wt.% Table 1. The chemical composition of mineral components, wt.%
Type
MgO Al2O3 SiO2
K2O CaO
Fe2O3 TiO2 SO3 LOI
TPP ash
2,49
23,89
42,88 0,48
4,6 7,95
0,11
0,66 16,9
Volcanic ash
0,20
13,57
73,67 6,00
1,79 1,52
2,85
-
0,40
Limestone flour 0,72
1,55
5,05
0,6 90,14
1,4
-
0,49 -
Quartz powder
6,32
14,99
73,83 1,83
0,6 0,97
1,32
0,14 - 1
Introduction Concrete is one of the oldest materials, but its potential and possibilities seem inex-
haustible (Murtazaev et al. 2016; Nesvetaev et al. 2018; Stelmakh et al. 2018), since at
all times of its existence and in the future this material will occupy a leading place
among a huge variety of building compositions. The active component of concrete is cement. It is known that varying finely dis-
persed mineral additives in its composition results in modern composite materials,
which properties will vary in wide ranges (Udodov 2015; Salamanova et al. 2017). The active component of concrete is cement. It is known that varying finely dis-
persed mineral additives in its composition results in modern composite materials,
which properties will vary in wide ranges (Udodov 2015; Salamanova et al. 2017). In accordance with GOST 31108-2003, granulated slag, fuel ashes, including acidic
or basic fly ash, microsilica, burnt clay, burnt shale, marl, quartz sand, etc. are used as
mineral components—main components of cement (Udodov 2015; Murtazaev et al. 2016). Various mineral additives can be used as auxiliary components of cement,
which will not significantly increase the water demand of cement and reduce durability
of concrete. In accordance with GOST 31108-2003, granulated slag, fuel ashes, including acidic
or basic fly ash, microsilica, burnt clay, burnt shale, marl, quartz sand, etc. are used as
mineral components—main components of cement (Udodov 2015; Murtazaev et al. 2016). Various mineral additives can be used as auxiliary components of cement,
which will not significantly increase the water demand of cement and reduce durability
of concrete. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 360–364, 2019. https://doi.org/10.1007/978-3-030-22974-0_87 361 Multicomponent Binders with Off-Grade Fillers 3
Results and Discussion To produce multicomponent binders, the additives under study were ground in VM-20
laboratory ball vibratory mill for 30 and 40 min. Figure 1 shows dependence of
specific surface of mineral additives on the grinding time. 0
100
200
300
400
500
600
700
800
900
1000
30 minutes
40 minutes
Specific surface, m2/kg
Grinding time
limestone
volcanic ash
TPP ash
quartz sand
Fig. 1. Specific surface of mineral components Fig. 1. Specific surface of mineral components To determine the optimal degree of saturation of Portland cement (PC) – mineral
powder (MP) system (PC:MP), samples were prepared from the proposed multicom-
ponent binder formulations and properties (Table 2). S.-A. Murtazaev et al. 362 Table 2. Properties of multicomponent binders (MCB) No. Mineral
Powder
PC:MP
Normal den-
sity,%
Setting time,
hour-min
Activity,
MPa
start
end
1
Limestone flour
70/30
25,5
2-05
3-00
35,8
2
60/40
26,8
2-15
3-20
30,4
3
Quartz powder
70/30
24,6
1-30
2-10
41,8
4
60/40
27,0
1-55
2-50
39,7
5
TPP Ash
70/30
26,4
2-10
3-15
34,1
6
60/40
28,1
2-25
3-35
28,2
7
Volcanic ash
70/30
25,2
1-35
2-15
42,6
8
60/40
26,5
2-05
3-00
40,3
9
–
100
25,0
2-20
3-40
48,0 The results of the studies showed that the most rational are the compositions of
binders using mineral powders of volcanic ash and quartz powder with a ratio of
70:30%, with a specific surface of 876 m2/kg and 650 m2/kg, respectively, with a
typical increase in the activity of the binder and a slight increase in normal thickness,
and 30% of portland cement are saved. Next, a concrete mixture with P2 mobility mark was produced, the samples were
subjected to heat and humidity treatment (HHT) in a steam chamber at 2 + 3 + 7 + 2 h
at an isothermal holding temperature of 80 °C. Table 3 shows the experimental
compositions and properties of the studied concretes. Table 3. The compositions of the studied concretes
No
composition
Mineral
powder
Consumption of
materials, kg/m3
Average density,
kg/m3
Compressive
strength, MPa
MCB-
70
ACS DS
B
After
HHT
Age 28
days
1
Limestone
flour
450
1100 680 220 2430
43,3
38,4
2
Quartz
powder
450
1100 680 210 2410
50,2
45,9
3
Volcanic
ash
450
1100 680 215 2415
52,1
46,5
4
TPP ash
450
1100 680 230 2420
37,7
35,9
5
PC
450
1100 680 200 2420
51,5
48,6
Note: PC – Portland cement; ACS – Alagir crushed stone fraction 5–20 mm; FS – fractionated
fine filled based on the sands of the Alagir and Chervlensk deposits. Table 3. The compositions of the studied concretes We established that the strength of concrete after HHT is 12% higher than the
indicators of the strength of concrete after 28 days of natural hardening. The use of
MCB-70 with volcanic ash showed the best results on the compressive strength of Multicomponent Binders with Off-Grade Fillers 363 concrete in comparison with other additives and slightly inferior to similar indicators of
control samples (Murtazaev et al. 2016; Stelmakh et al. 2018). Table 2. Properties of multicomponent binders (MCB) The study of operational
characteristics (Table 4) showed that the indicators of these properties depend on the
composition of the MCB-70 and its activity, as well as on the type and value of the
porosity of the material. Table 4. Operational properties of concrete using MCB-70
Indicators
Mineral powder
Limestone
flour
Volcanic
ash
TPP
ash
Quartz
powder
MCB-70 activity, MPa
35,8
42,6
34,1
41,8
Compressive strength, MPa
38,4
46,5
35,9
45,9
Flexural strength, MPa
4,1
4,9
3,8
4,4
Porosity, %
9,7
7,6
12,4
6,9
Frost resistance, cycle
F300
F350
F200
F350
Pressure, MPa
1,4
1,8
1,2
1,8
Water absorption, %
4,2
3,5
5,2
3,6
Water resistance, Kr - softening
coefficient
0,79
0,89
0,63
0,90 Table 4. Operational properties of concrete using MCB-70 4
Conclusions Multicomponent binders based on mineral powders of natural and man-made origin
allow to obtain high-quality concrete of class of strength B30-40, including for high
monolithic construction. References Udodov SA (2015) Re-introduction of plasticizer as a tool for controlling the mobility of concrete
mix. In: Proceedings of the Kuban State Technological University, no 9, pp 175–185 Murtazaev S-Y, Salamanova MS, Bisultanov RG, Murtazaeva TS-A (2016) High-quality
modified concretes using a binder based on a reactive active mineral component. Stroitelnyematerialy, no 8, pp 74–80 Nesvetaev G, Koryanova Y, Zhilnikova T (2018) On effect of superplasticizers and mineral
additives on shrinkage of hardened cement paste and concrete. In: MATEC Web of
Conferences 27, Cep, 27th R-S-P Seminar, Theoretical Foundation of Civil Engineering
(27RSP), TFoCE 2018, p 04018 Stelmakh SA, Nazhuev MP, Shcherban EM, Yanovskaya AV, Cherpakov AV (2018) Selection
of the composition for centrifuged concrete, types of centrifuges and compaction modes of
concrete mixtures. In: Kim Y-H, Parinov IA, Chang S-H (ed) Physics and Mechanics of New
Materials and Their Applications (PHENMA 2018) Abstracts & Schedule, p 337 Salamanova M, Khubaev M, Saidumov M, Murtazaeva T (2017) Self-consolidating concretes
with materials of the Chechen Republic and neighboring regions. Int J Environ Sci Educ 11
(18):12719–12724 364 S.-A. Murtazaev et al. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction The modern materials science and construction now deal with an important national
economic and engineering problem: development of efficient technologies for pro-
ducing high-strength monolithic concretes through the integrated use of technogenic
raw materials to obtain secondary raw materials for concrete, while eliminating the
enormous environmental damage caused by waste “cemeteries” (Bazhenov et al. 2006;
Salamanova et al. 2017; Lesovik et al. 2012; Murtazaev et al. 2009; Kaprielov et al. 2018) in Russia and the world and, in particular, for the Chechen Republic, considering
construction of 435-m high-rise complex «Akhmat-Tower» in Grozny City (Udodov
2015; Koryanova et al. 2018; Salamanova and Murtazaev 2018). High-Quality Concretes for Foundations
of the Multifunctional High-Rise Complex
(MHC) «Akhmat Tower» S.-A. Murtazaev1,2,3, M. Saydumov1(&), A. Alaskhanov1,
and M. Nakhaev4 1 Millionshchikov Grozny State Oil Technical University, Grozny, Russia
saidumov_m@mail.ru
2 Ibragimov Complex Research Institute, RAS, Grozny, Russia
3 Academy of Sciences of the Chechen Republic, Grozny, Russia
4 Chechen State University, Grozny, Russia 1 Millionshchikov Grozny State Oil Technical University, Grozny, Russia
saidumov_m@mail.ru
2 Ibragimov Complex Research Institute, RAS, Grozny, Russia
3 Academy of Sciences of the Chechen Republic, Grozny, Russia
4 Chechen State University, Grozny, Russia Abstract. The paper presents results of studies of monolithic concrete mixes
and concretes produced with the integrated use of local natural and technogenic
raw materials, including waste scrap and crushed bricks. We developed optimal
compositions of monolithic concretes and studied their technological and
physical-mechanical properties. Keywords: High-strength concrete High-quality concrete mix
Filled binder Technogenic wastes Mineral technogenic filler
Monolithic concrete Keywords: High-strength concrete High-quality concrete mix
Filled binder Technogenic wastes Mineral technogenic filler
Monolithic concrete Keywords: High-strength concrete High-quality concrete mix
Fill d bi d
T
h
i
t
Mi
l t
h
i fill Keywords: High-strength concrete High-quality concrete mix
Filled binder Technogenic wastes Mineral technogenic filler © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 365–368, 2019.
https://doi.org/10.1007/978-3-030-22974-0_88 2
Methods and Materials Our experimental studies used local additive-free Portland cement of PC 500 D0 grade
as a binder. Natural sand from the Chervlensky deposit of the Chechen Republic was
used as a fine filler. Local gravel of 5–20 mm fractions from the Argunsky and Ser-
novodsky deposits of the Chechen Republic and imported crushed stone of 5–20 mm S.-A. Murtazaev et al. 366 fraction from granite-diabase rocks of the Alagirsky deposit of the Republic of North
Ossetia-Alania were used as a coarse filler. As plastifying agents in accordance with GOST 24211-2008 “Additives for con-
crete. General technical requirements” modern additives of various manufacturers of
building chemicals (POLYPLAST, TOKAR, etc.) were used. The raw materials for the production of dispersed technogenic mineral fillers
(DTMF) were local materials, mainly technogenic, namely concrete scrap, crushed
ceramic bricks (CCB), ash and slag mixture from the Grozny heat and power plant
(HPP) and very small non-conditioned quartz sands ones were used in comparative
tests. All the DTMFs were ground for 5 min in MV-20-EX laboratory vibratory ball mill
with a loading volume of 5–6 L to obtain a specific surface of 450–600 m2/kg. 3
Results and Discussion The Filled Binder (FB) formulation was developed and investigated with activity 60–
71 MPa with concrete scrap and CCB fillers with ratio 70:30%. The proportion of the
mixture of filler in FB was 25 and 40% by weight of the binder. Due to the fact that for designing the underground part of the Akhmat Tower
multifunctional complex, concrete of different strength classes (B40, B75-B80) was
laid, the task was to develop high-quality concrete mixes (HCM), starting from the
middle B40-B50 classes and ending with high-strength concrete of B80-B90 classes,
with the integrated use of local raw materials, including with technogenic nature. Fig. 1. Temperature change and stress characteristic in fresh concrete with limited deformation:
rCS - internal compressive stresses; rCST - the same, tensile; tIN - initial temperature of concrete
mixture; tmax - the same, maximum; tAT - ambient temperature (air); I, II, … V - stages (periods)
of process of heat dissipation of the concrete mix in time; curve 1 - kinetics of heat dissipation of
concrete with PC; curve 2 - the same with FB Fig. 1. Temperature change and stress characteristic in fresh concrete with limited deformation:
rCS - internal compressive stresses; rCST - the same, tensile; tIN - initial temperature of concrete
mixture; tmax - the same, maximum; tAT - ambient temperature (air); I, II, … V - stages (periods)
of process of heat dissipation of the concrete mix in time; curve 1 - kinetics of heat dissipation of
concrete with PC; curve 2 - the same with FB 367 High-Quality Concretes for Foundations In massive building constructions, such as the Akhmat Tower MFC base plate,
because of their large dimensions, as a rule, the heat from cement hydration is slowly
released into the air or into adjacent structural elements, as a result of which the core of
the monolithic element heats up much faster and stronger than the shell. Therefore, we
investigated thermophysical processes of the developed compositions. Figure 1 schematically shows the dependence of temperature and voltage due to
external pressure generated by this concreting technology. The dependence of heat release curves in time is conventionally divided into 5
stages (Table 1). Table 1. References Bazhenov YM, Demyanova BC, Kalashnikov VI (2006) Modified high-quality concretes. Publishing House of the Association of Construction Universities, Moscow, 368 p Kaprielov SS, Sheinfeld AV, Al-Omais D (2018) Experience in the production and quality
control of high-strength concrete on the construction of the high-altitude complex “OKO” in
Moscow-City International Business Center. Industrial and Civil Construction, no 1, pp 18–
24 oryanova YI, Rezantsev NE, Shumilova AS (2018) Materials and structures used in th
construction of high-rise buildings - from tradition to innovation. Alley Sci 6(4):95–99 construction of high-rise buildings - from tradition to innovation. Alley Sci 6(4):95–99
Lesovik BC, Murtazaev SAY, Saydumov MS (2012) Construction composites based on
screenings of crushing of concrete scrap and rocks. MUP “Typography”, Groznyy, 192 p
Murtazaev S-AY, Bataev DK-S, Ismailova ZK (2009) Fine-grained concretes based on fillers
from secondary raw materials. Comtechprint, Moscow, 142 p esovik BC, Murtazaev SAY, Saydumov MS (2012) Construction composites based on
screenings of crushing of concrete scrap and rocks. MUP “Typography”, Groznyy, 192 p y
(
)
p
screenings of crushing of concrete scrap and rocks. MUP “Typography”, Groznyy, 192 p
Murtazaev S-AY, Bataev DK-S, Ismailova ZK (2009) Fine-grained concretes based on fillers
from secondary raw materials. Comtechprint, Moscow, 142 p Murtazaev S-AY, Bataev DK-S, Ismailova ZK (2009) Fine-grained concretes based on fi
from secondary raw materials. Comtechprint, Moscow, 142 p alamanova M, Khubaev M, Saidumov M, Murtazayeva T (2017) Int J Environ Sci Educ 1
(18):12719–12724 Salamanova MS, Murtazaev SAY (2018) Clinker-free binders based on finely dispersed mineral
components. In: Collection: Ibausil Conference Proceedings, pp 707–714 Udodov SA (2015) Re-introduction of plasticizer as a tool for controlling the mobility of concrete
mix. In: Proceedings of the Kuban State Technological University, no 9, pp 175–185 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. 3
Results and Discussion The main stages (periods) of the process of heat dissipation in time of concrete mixes
with various binders
Stage
№
Duration, h
Description
With
PC
With
FB
1
0–2
0–8
The initial stage without raising the temperature of the concrete mix
(dormant period). This period is significantly increased due to the use
of surface-active substances (surfactants) in the composition of FB,
hardening retarders, etc. 2
2–6
8–15
Temperature increase due to hydration of the binder, no measurable
stress, because in the plastic concrete, thermal expansions are
converted to relative compression. At the end of this stage, the
temperature is referred to as “the first temperature at zero stress” t01
3
6–13
15–24
Further heating of the concrete, the strength of the concrete increases
and a compressive stress is formed, partially decreasing because of
relaxation. Stage III ends when the maximum temperature tmax is
reached
4
13–
24
24–72
Heat transfer prevails: the temperature of concrete and compressive
stress in concrete decrease, a part of compressive stress decreases
because of relaxation. The “second temperature at zero stress” t02 is
reached, which significantly exceeds t01 in cooling rate and age of
concrete
5
24–
72
72–144 Further cooling and increasing tensile stress, which are partially
reduced due to relaxation. If the tensile stress reaches the tensile
strength of the concrete under tension (at ΔTКРИТ) through cracks are
formed Table 1. The main stages (periods) of the process of heat dissipation in time of concrete mix
with various binders According to calculations (Kaprielov et al. 2018), the temperature difference
between the upper surface layers of concrete slab and outside air DT1 should be no
more than 20 °C, and the difference between the side layers DT2 - no more than 30 °C. S.-A. Murtazaev et al. 368 4
Conclusions Thus, analysis of data confirms the effectiveness of the use of FB with DTMF in high-
quality concretes used for concreting massive structures. We established that the peak
of the maximum heat release tmax from the exotherm of cement in (massive) concrete
on HB was reduced by 30–35% in comparison to the concrete with PC (from 70–75 °C
to 50–55 °C). 1
Introduction A great interest for application of the secondary product of crushing concrete scrap of
dismantling buildings and structures, according to the authors of the paper, is related to
the possibility of its application as a fine-milled mineral component in filled binders,
based on the use of which it is possible to produce high-strength concrete, including
monolithic high-rise construction (Bazhenov et al. 2011; Batayev et al. 2017;
Murtazaev et al. 2009). Designing High-Strength Concrete Using
Products of Dismantling of Buildings
and Structures T. Murtazaeva1(&), A. Alaskhanov1, M. Saidumov1,
and V. Hadisov1,2 T. Murtazaeva1(&), A. Alaskhanov1, M. Saidumov1,
and V. Hadisov1,2 1 Millionshchikov Grozny State Oil Technical University, Grozny, Russia
tomamurtazaeva@mail.ru
2 Ibragimov Complex Research Institute, RAS, Grozny, Russia Abstract. The paper presents an analysis of experience of using the products of
dismantling of buildings and structures, the technology of recycling of sec-
ondary raw materials to produce secondary raw materials for concrete. We
presented results of tests of heavy concrete based on filled binders using the
products of processing of concrete and brick scrap. Keywords: Building demolition products Concrete scrap Brick scrap
Ecology Recycling Secondary aggregate Fine ground aggregate filled with binder References If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 4
Conclusions The optimal formulations of highly mobile concrete mixtures were designed using local
natural and technogenic raw materials with a grade of P5 cone sediment and persistence
for more than 8 h to obtain HSC classes of compressive strength up to B60-B80 with
unique performance properties. 2
Methods and Materials The following materials were used as raw materials for concrete: natural sand from the
Chervlenskoye deposit, crushed stone from gravel of 5–20 mm fractions from the
Argunsky and Sernovodsky deposits, imported crushed stone of a 5–20 mm fraction
from granite-diabase rocks of the Alagirsky deposit of the Republic of North Ossetia-
Alania, local non-additive portland cement of brand PC 500 D0, plastification additives
Polyplast and D-5. Polyplast and D-5. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 369–371, 2019. https://doi.org/10.1007/978-3-030-22974-0_89 © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 369–371, 2019. https://doi.org/10.1007/978-3-030-22974-0_89 370 T. Murtazaeva et al. 3
Results and Discussion To obtain the optimal formulations of high-strength concrete (HSC) with the integrated
use of local raw materials, including of technogenic nature, compositions of filled
binders (FB) with fine-milled mineral filler of technogenic nature (MFTN) of HB-75:25
and HB-60:40grades, allowing to obtain high-strength cement stone with noticeably
smaller pores and less shrinkage (Murtazaev et al. 2009; Udodov 2015; Murtazaev and
Salamanova 2018). From the test results of HSC based on FB, it can be seen that the dynamics of the
strength of concrete on FB are noticeably different from the dynamics of the growth of
concrete strength on Portland cement. It was established that the process of durability of concrete on FB at an early age
(1–3 days) is accelerated by 1.5–2 times. So, concrete on FB at the age of 1 day has a
strength of about 33–36% of the designed, and 3 days old - this indicator reaches 70%. 7-day strength of concrete, produced using FB, is about 85–90% of the designed, which
is significantly higher than traditional compositions on ordinary Portland cement. These
indicators for concrete on Portland cement at the age of 1, 3 and 7 days are about 24, 35
and 70% of the designed strength, respectively. References Bazhenov YM, Bataev DK-S, Mazhiev KN (2011) Fine-grained concretes from recycled
materials for the repair and restoration of damaged buildings and structures. FE “Sultanbe-
gova Kh.S.”, Grozny, 342 p Bazhenov YM, Bataev DK-S, Mazhiev KN (2011) Fine-grained concretes from recycled
materials for the repair and restoration of damaged buildings and structures. FE “Sultanbe-
gova Kh.S.”, Grozny, 342 p Batayev DK-C, Saidumov MS, Murtazaeva TS-A (2017) Recipes of high-strength concretes on
technogenic and natural raw materials. In: Materials of the All-Russian Scientific and
Practical Conference Dedicated to the 60th Anniversary of the Building Department of
Millionshchikov GSTU, 12–13 October 2017, Bisultanova P.Sh., Grozny, pp 109–117 Murtazaev S-AY, Bataev DK-S, Ismailova ZK (2009) Fine-grained concretes based on fillers
from secondary raw materials. Comtechprint, Moscow, 142 p Udodov SA (2015) Re-introduction of plasticizer as a tool for controlling the mobility of concrete
mix. In: Proceedings of the Kuban State Technological University, no 9, pp 175–185 mix. In: Proceedings of the Kuban State Technological University, no 9, pp 175–185
Murtazaev SAY, Salamanova MS (2018) Clinker-free binders based on finely dispersed mineral
components. In: Collection: Conference Proceedings, pp 707–714 Murtazaev SAY, Salamanova MS (2018) Clinker-free binders based on finely dispersed mineral
components. In: Collection: Conference Proceedings, pp 707–714 371 Designing High-Strength Concrete Using Products of Dismantling © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 372–375, 2019.
https://doi.org/10.1007/978-3-030-22974-0_90 Estimation of Rheo-Technological Effectiveness
of Polycarboxylate Superplasticizer in Filled
Cement Systems in the Development
of Self-compacting Concrete for High-Density
Reinforced Building Constructions T. Nizina(&), A. Balykov, V. Volodin, and D. Korovkin Department of Building Structures, Ogarev Mordovia State University,
Saransk, Russia
nizinata@yandex.ru Abstract. Analysis of rheo-technological effectiveness of the polycarboxylate
superplasticizer Melflux 5581 F in the microcalcite-filled cement systems was
performed. Optimal quantities of polycarboxylate superplasticizer and carbonate
filler were determined, which allow obtaining highly mobile cement-mineral
suspensions at reduced water content, which is an important step in the devel-
opment of self-compacting concrete mixtures. Keywords: Cement-mineral suspension Rheology Efficiency
Superplasticizer Microcalcite Self-compacting concrete Designing High-Strength Concrete Using Products of Dismantling Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches To prepare suspensions, Portland cement CEM I 32.5R (C) by Mordovtsement PJSC
(GOST 31108) was used. The mineral part included a carbonate filler from KM100
microcalcite (MKM) by Polipark LLC with the dosage of 0300% of Portland cement
weight (075% of solid phase weight) with the variability pitch of 100%. Melflux
5581 F (SP) polycarboxylate superplasticizer by BASF Construction Solutions
(Trostberg, Germany) was used as a plasticizer. The study was carried out with the fixed water-solid ratio W/S = 0.15 with the
varying factors being: – ratio MKM/C, x1 = 03.0 relative units; – ratio SP/(C + MKM), x2 = 01.5%. – ratio SP/(C + MKM), x2 = 01.5%. 1
Introduction Currently, there is an active growth in the area of concrete development, which the
world technological community clearly classifies as cement composites of the new
generation with high strength, workability, volume stability and durability (Collepardi
2006; Nawy 2001; Nizina and Balykov 2016; Nizina et al. 2017; Sivakumar et al. 2014; Tran and Kim 2017). A special place among the concretes of the new generation
is occupied by Self-Compacting Concrete (SCC) – Selbstverdichtender Concrete (SVB,
German), Betonautoplacant (BAP, French), which are currently fairly widespread
abroad. This term, proposed in 1986 by the Japanese professor H. Okamura (Okamura
and Ouchi 2003), combines concrete mixtures with high workability characteristics
(standard cone flow over 55–60 cm at water-to-cement ratio reduced to 0.35–0.4 or
less), which are due to the high deformability of the suspension matrix, along with its
high resistance to segregation or separation during movement. A number of papers experimentally proved that one of the basic principles for
producing self-compacting concrete mixtures is the presence of a significant amount of
dispersed micro-particles of cement or mineral fillers (mainly 1–100 microns in size) in
their formulation, which, together with Portland cement, increase the volume of a
dispersion-water suspension. At the same time, not all dispersed fillers are capable of
providing a higher flowability in suspension with a superplasticizer (SP) as compared Estimation of Rheo-Technological Effectiveness 373 with cement suspensions. The research (Kalashnikov et al. 2014) shows that carbonate
rocks (limestone, marble, dolomite), which include particles with a significant pro-
portion of positively charged active centers, are the most compatible with anionic
superplasticizers. Thus, the generate of self-compacting concrete mixtures must begin with the
development of rheological active formulation of the filled cement binders, which
allow, when mixing together with the superplasticizer, to form aggregate resistant
suspensions, which have a high concentration of the solid phase, low values of the
shear stress limit and plastic viscosity at high gravitational fluidity under its own
weight. At the same time, the efficiency of plasticizers in such systems will depend on
many factors – addition procedure and optimal quantity of the plasticizer, rheological
activity of the fillers used, etc. The purpose of the study is to determine the optimal quantity of polycarboxylate
superplasticizer and carbonate filler in cement-mineral suspensions when developing
self-compacting concrete mixtures. 3
Results and Discussion The study results were used to develop an experimentally statistical (ES) model
describing the changes in the flow diameter of cement-carbonate suspensions (Portland
cement + microcalcite) [DHC
f
, mm] from the Hegermann cone from the content of the
varying factors x1 and x2: DHC
f
¼ 288:9 þ 52:9 x1 þ 64:8 x2 þ 19:8 x1 x2 51:33 x2
1
89:3 x2
2 þ 37:97 x3
1 þ 32:34 x3
2 59:06 x1 x2
2
15:19 x2
1 x2 þ 31:64 x2
1 x2
2:
ð1Þ ð1Þ Using the polynomial (1), isolines have been built, which reflect the changes in the
flow diameter of cement-carbonate suspensions from the Hegermann cone depending
on the content of microcalcite and Melflux 5581 F superplasticizer (Fig. 1). It has been T. Nizina et al. 374 found that for the constant water/solid ratio of W/S = 0.15 relative units, an increase in
the dosage of the superplasticizer and the mineral filler (microcalcite) causes a sig-
nificant increase in the flow diameter of the cement-mineral suspension. Fig. 1. Changes in the flow diameter of cement-carbonate suspensions from the Hegermann
cone depending on the superplasticizer dosage and microcalcite filling degree Fig. 1. Changes in the flow diameter of cement-carbonate suspensions from the Hegermann
cone depending on the superplasticizer dosage and microcalcite filling degree Analysis on Fig. 1 shows that cement suspensions without microcalciteat the
specified water content W/S = 0.15 begin to flow under the action of gravity and the
value of the Hegermann cone flow diameter DHC
f
= 100135 mm with superplasticizer
quantities exceeding 0.10.5% the mass of the solid phase. However, even an increase
in the superplasticizer content to 1.01.5% does not allow achieving self-compacting
of suspensions (DHC
f
¼ 160 210\280 mm). It was determined (Fig. 1) that the level of rheo-technological indices specified for
self-compacting suspensions (DHC
f
280 mm) is achieved at the microcalcite content
x1 = 0.823.0 relative units (82300% of the Portland cement mass or 4575% of the
solid phase mass (C + MKM)) and the quantity of Melflux 5581 F superplasticizer
x2 = 0.451.5% of the solid phase mass, wherein when decreasing the indicator x1, an
increase in the indicator x2 in the specified ranges ((x1 = 3.0 relative units;
x2 = 0.45%) ! 3
Results and Discussion (x1 = 0.82 relative units; x2 = 1.5%)) is required. 4
Conclusions The analysis of research results found the optimal levels of the varying factors that
allow reaching the self-compacting of suspensions for the flow diameter from the
Hegermann cone above 280 mm and the water/solid ratio of 0.15 relative units: the
dosage of Melflux 5581 F superplasticizer is 0.51.0% of the solid phase weight; the
filling degree of suspension with microcalcite is at least 105% of the Portland cement
weight. Estimation of Rheo-Technological Effectiveness 375 Acknowledgements. The reported study was funded by RFBR according to the research project
№18-29-12036. №18-29-12036. References Collepardi M (2006) The New Concrete. Grafiche Tintoretto, Villorba Kalashnikov VI, Moskvin RN, Belyakova EA, Belyakova VS, Petukhov AV (2014) High-
dispersity fillers for powder-activated concretes of new generation. Syst Methods Technol 2
(22):113–118 Nawy EG (2001) Fundamentals of High-Performance Concrete. Wiley, New York Nizina TA, Balykov AS (2016) Experimental-statistical models of properties of modified fiber-
reinforced fine-grained concretes. Mag Civil Eng 2:13–25. https://doi.org/10.5862/MCE.62.2 Nizina TA, Ponomarev AN, Balykov AS, Pankin NA (2017) Fine-grained fibre concretes
modified by complexed nanoadditives. Int J Nanotechnol 14:665–679. https://doi.org/10. 1504/IJNT.2017.083441 Okamura H, Ouchi M (2003) Self-compacting concrete. J Adv Concr Technol 1:5–15 Sivakumar N, Muthukumar S, Sivakumar V, Gowtham D, Muthuraj D (2014) Experimental
studies on high strength concrete by using recycled coarse aggregate. Res Inventy: Int J Eng
Sci 4:27–36 Tran NT, Kim DJ (2017) Synergistic response of blending fibers in ultra-high-performance
concrete under high rate tensile loads. Cement Concr Compos 78:132–145 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction The production of photocatalytic composite materials (PCM) of the “TiO2 – SiO2”
system is aimed at increasing the efficiency of photocatalytic decomposition of pol-
lutants (Arai et al. 2006; Guo et al. 2016).The peculiarities of physical and chemical
interaction of siliciferous and titanium-containing components in the synthesis and use
of PCM directly affect its photocatalytic activity. In this regard, it is important to study
the influence of the properties of siliciferous raw materials on the final characteristics of
PCM. Parameters of Siliciferous Substrate
of Photocatalytic Composition Material
as a Factor of Its Efficiency Y. Ogurtsova(&), E. Gubareva, M. Labuzova, and V. Strokova Department of Materials Science and Technology,
Belgorod State Technological University named after V.G. Shoukhov,
Belgorod, Russia
ogurtsova.y@yandex.ru Abstract. The article presents the results of the determination pf the properties
of the photocatalytic composite material (PCM) of the “TiO2 – SiO2” system
synthesized by the sol-gel method. The characteristics of siliciferous raw
material - diatomite and silica clay, as substrates in the composition of PCM -
mineral composition, microstructural features, composition and concentration of
active centers on its surface are determined. The dependences of the elemental
composition of the surface, the features of the microstructure and photocatalytic
activity of PCM on the properties of siliciferous raw material are found. The
research shows that the use of diatomite makes it possible to obtain PCM with
better characteristics, which is caused by a higher content of the amorphous
phase, a more developed and chemically active surface of the particles. Keywords: Siliciferous raw material Titanium dioxide
Sol-gel Photocatalysis Microstructure Activity Keywords: Siliciferous raw material Titanium dioxide
Sol-gel Photocatalysis Microstructure Activity © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 376–380, 2019.
https://doi.org/10.1007/978-3-030-22974-0_91 2
Methods and Approaches As a siliciferous raw material and as a substrate in the composition of the photocatalytic
composite material, the Diasil diatomaceous fine dispersed powder (specific surface
Ss = 1.39 m2/g) was used (Diamix, Ulyanovsk region, Russia); fine-ground silica clay
(Ss = 1.08 m2/g) (Alekseevskii deposit, Mordovia, Russia) were used. The determi-
nation of the mineral composition of siliciferous raw materials was carried out using an Parameters of Siliciferous Substrate of PCM 377 ARL 9900 WorkStation X-ray fluorescence spectrometer. The peculiarities of
microstructure and elemental composition of the surface were studied with the help of
TESCAN MIRA 3 LMU high resolution scanning electron microscope. The acid-base
characteristics of the surface of siliciferous raw materials were studied using the
indicator method (Nechiporenko 2017; Nelyubova et al. 2018). The production of composite material of the TiO2 – SiO2 system was obtained by
the sol-gel method using a titanium-containing organic precursor—titanium butoxide
Ti (OC4H9)4 (TBT) (TU 6-09-2738-89, “PROMHIMPERM”, Russia). It was dissolved
in ethanol, and then the siliciferous substrate (SS) material was introduced into the
resulting solution, in a ratio of “TBT/SS” - 4/1. After stirring it on a magnetic mixer,
the material was dried and burned at 550 °C. Then, the tablets were prepared from the obtained materials of the “TiO2 – SiO2”
system. White cements CEM I 52, 5 R (Adana, Turkey) was used as a binder. The ratio
TiO2 – SiO2/cement is 1.3/1. The photocatalytic activity was determined using the
photocatalytic decomposition method of the organic pigment Rhodamine B (Rho-
damine B, C28H31ClN2O3). The pigment was applied to the tablets at a concentration of
4 ∙10−4 mol/l. The samples were kept for 4 and 26 h under ultraviolet radiation (UV-
A, 1.1 ± 0.1) W/m2). The evaluation of color change, as an indicator of the effec-
tiveness of self-cleaning of the surface, was carried out according to the Lab color
space (coordinate a) using software (Guo et al. 2016). 3
Results and Discussion Diatomite is a sedimentary biogenic rock consisting of microscopic siliciferous shells
of algae (diatoms) with a valve size of 5–200 mcm. The presence of nanoscale pores
and elements is shown on the Fig. 1a. Silica clay is of sedimentary biogenic and
chemogenic origin, composed mainly of opal-cristobalite silica particles with a size of
less than 5 mcm. It is a microporous rock and the content of organic fragments is
insignificant (Fig. 1b). The mineral composition of the raw material is similar; a higher
content of the amorphous phase is found in the composition of diatomite (Table 1). Fig. 1. Microstructure of siliciferous raw materials: a – diatomite, b – silica clay Fig. 1. Microstructure of siliciferous raw materials: a – diatomite, b – silica clay Y. Ogurtsova et al. 378 Table 1. Mineral composition of siliciferous raw material
Siliciferous raw
material
a-Quartz
Cristobalite
low
Tridimite low Illite 2M1
Albite
Amorphous
Diatomite
9.35 ± 0.72 40.57 ± 4.22
4.21 ± 0.90 2.44 ± 0.89 0.53 ± 0.20 42.90
Silica clay
17.36 ± 0.72 31.63 ± 1.42 14.63 ± 1.31 3.42 ± 0.65 0.54 ± 0.12 32.42 Table 1. Mineral composition of siliciferous raw material The presence of a high concentration of acid sites characterized by proton acidity
(Brønsted) is noted on the surface of diatomite (Table 2). Table 2. The composition and concentration of active centers on the surface siliciferous raw
materials (q ∙10 −3, mEq/g)
Siliciferous raw
material
pKa
−0.29
Lewis bases
+0.80
+7.15
+12.00
+12.80
Bronsted acids
Bronsted bases
Diatomite
53.11
490.43
54.09
47.95
55.80
Silica clay
115.90
No data
20.51
98.68
13.41 able 2. The composition and concentration of active centers on the surface siliciferous raw
aterials (q ∙10 −3, mEq/g) The photocatalytic activity of PCM based on diatomite (Table 3) is high and close
to the control specimen – the industrial nano-sized Aeroxide TiO2 P25 photocatalyst. Table 3. Elimination of Rhodamine B, %
Time
Aeroxide
TiO2 P25
Diatomite
TiO2−SiO2
(diatomite)
Silica
clay
TiO2−SiO2
(Silica clay)
4 hours
29
9
29
1
5
26 hours
91
61
86
43
68 Table 3. Elimination of Rhodamine B, % The analysis of the peculiarities of the microstructure and elemental composition of
the surface of synthesized PCM based on diatomite (Fig. 2a) and silica clay (Fig. 2b)
shows that the silica particles are partially covered with titanium-containing new for-
mations. 4
Conclusions The siliciferous raw material differs in morphology, concentration of acid-base centers
and content of the amorphous phase: the surface of the diatomite is more developed,
characterized by a high concentration of proton acid centers; it has a higher content of
the amorphous phase in its composition. As a result, the photocatalytic activity of PCM
synthesized on the basis of diatomite is higher by 20% in comparison with PCM on the
basis of silica clay. In order to improve the efficiency, it is advisable to consider the
possibility of pre-activation of silica clay, which will allow using this waste (by-
product) rock to produce modern self-cleaning materials. Acknowledgements. The research was carried out with financial support from Russian Science
Foundation grant (project №19-19-00263). 3
Results and Discussion The surface of PCM particles based on diatomite is more developed; the
distribution of the titanium-containing phase is more even. Parameters of Siliciferous Substrate of PCM 379 Fig. 2. Microstructure of PCM on the basis of: a – diatomite, b – silica clay (with mapping by
elements) Fig. 2. Microstructure of PCM on the basis of: a – diatomite, b – silica clay (with mapping by
elements) Acknowledgements. The research was carried out with financial support from Russian Science
Foundation grant (project №19-19-00263). Guo M-Z, Maury-Ramirez A, Poon CS (2016) Self-cleaning ability of titanium dioxide clear
paint coated architectural mortar and its potential in field application. J Clean Prod 112:3583–
3588 Arai Y, Tanaka K, Khlaifat AL (2006) Photocatalysis of SiO2-loaded TiO2. J Mol Catal A: Chem
243:85–88 Properties Improvement
of Metakaolin-Zeolite-Diatomite-Red Mud
Based Geopolymers F. Rocha1(&), C. Costa1, W. Hajjaji1, S. Andrejkovičová1,
S. Moutinho2, and A. Cerqueira1 1 GeoBioTec, Geosciences Department, University of Aveiro,
3810-193 Aveiro, Portugal
tavares.rocha@ua.pt
2 RISCO, Civil Engineering Department, University of Aveiro,
3810-193 Aveiro, Portugal Abstract. Addition of pozzolanic materials increases the mechanical charac-
teristics of construction materials and contributes towards a higher durability. Metakaolin is an artifical pozzolan obtained by calcination of kaolinitic clays at
an adequate temperature. Geopolymers are inorganic materials from mineral
origin, composed of a precursor, an alkaline activator and a solvent. New
geopolymer formulations were designed by sodium silicate/NaOH/KOH acti-
vation of metakaolin, zeolites, diatomites and red mud mixtures. The effects of
source materials on the microstructure and mechanical properties were studied. Mineralogical and chemical compositions were assessed as well as microstruc-
ture, specific surface area, compressive strength and adsorption. In general,
incorporation of red mud, zeolite filler and diatomites to metakaolin in medium of
alkali activators of low concentration provided formation of more eco-friendly
materials with high mechanical resistances and water treatment capabilities. Keywords: Geopolymers Metakaolin Alkaline activation
Additives formulations Keywords: Geopolymers Metakaolin Alkaline activation
Additives formulations Keywords: Geopolymers Metakaolin Alkaline activation
Additives formulations References Arai Y, Tanaka K, Khlaifat AL (2006) Photocatalysis of SiO2-loaded TiO2. J Mol Catal A: Chem
243:85–88 Guo M-Z, Maury-Ramirez A, Poon CS (2016) Self-cleaning ability of titanium dioxide clear
paint coated architectural mortar and its potential in field application. J Clean Prod 112:3583–
3588 Guo M-Z, Maury-Ramirez A, Poon CS (2016) Self-cleaning ability of titanium dioxide clear
paint coated architectural mortar and its potential in field application. J Clean Prod 112:3583–
3588 Nechiporenko AP (2017) Donor-acceptor properties of surface of solid-phase systems. Indicator
method, 1st edn. Lan, St. Petersburg Nelyubova VV, Sivalneva MN, Bondarenko DO, Baskakov PS (2018) Study of activity of
polydisperse mineral modifiers via unstandardized techniques. J Phys: Conf Ser 118:012029 Nelyubova VV, Sivalneva MN, Bondarenko DO, Baskakov PS (2018) Study of activity of
polydisperse mineral modifiers via unstandardized techniques. J Phys: Conf Ser 118:012029 Nelyubova VV, Sivalneva MN, Bondarenko DO, Baskakov PS (2018) Study of activity of
polydisperse mineral modifiers via unstandardized techniques. J Phys: Conf Ser 118:012029 380 Y. Ogurtsova et al. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 381–384, 2019.
https://doi.org/10.1007/978-3-030-22974-0_92 1
Introduction Addition of pozzolanic materials increases the mechanical characteristics of con-
struction materials and contributes towards a higher durability. Metakaolin is an
artifical pozzolan obtained by calcination of kaolinitic clays at an adequate temperature. Metakaolin is the structurally disordered product obtained following the dehydroxy-
lation of kaolin, more precisely of its essential component kaolinite - Al2Si2O5(OH)4. Geopolymers are inorganic materials from mineral origin, composed of a precursor,
an alkaline activator and a solvent. The process of geopolymerization involves a
chemical reaction that takes place in an alkaline medium, resulting in the formation of
inorganic polymers that have silicon and aluminum as main constituents (Si+Al),
connected by oxygen ions. The solution becomes alkaline using activators such as
sodium and potassium hydroxides and/or sodium and potassium silicates. 382 F. Rocha et al. Depending on the composition, geopolymer characteristics can be attained in terms
of physical, chemical and mechanical performance (Mackenzie and Welter 2014). Moreover, geopolymers have the advantage to be possibly formulated from a wide
range of aluminosilicate minerals. 2
Methods and Approaches 383 Properties Improvement of Metakaolin-Zeolite-Diatomite-Red Mud Properties Improvement of Metakaolin-Zeolite-Diatomite-Red Mud 3
Results and Conclusions Performed analyses confirm that zeolite particles are responsible for higher amount of
crystalline phases producing more compact and firm microstructure of blended
geopolymers relative to that of reference MK100. SEM analysis reveals that incor-
poration of 50 wt% of zeolite to metakaolin geopolymer (MK50) leads to the denser
geopolymer matrix related to microstructure when compared to 100 wt% metakaolin in
reference geopolymer (MK100). Dense microstructure of MK50 manifests in its con-
siderably lower water adsorption and specific surface area values. All geopolymers containing zeolite show increase in compressive strength com-
pared to pure metakaolin one, with optimal ratio metakaolin precursor/zeolite filler
50:50 providing the highest compressive strength (8.8 MPa at 28 days). The adsorption
of heavy metals increases as the amount of metakaolin in the structure increases. Diatomite enriched formulations showed different evolutions according to the Na or
K activator; those K activated showed higher compressive strength. The well-known geopolymer composition (amorphous phase predominant over
residual quartz, illite and anatase) is slightly affected by the red mud, despite it provides
aluminum and iron oxides and oxy-hydroxide (hematite, goethite, gibbsite and boeh-
mite). These phases, in fact, are to a limited extent involved in the geopolymerization
process and the alkaline aluminosilicate amorphous phase is about 90 wt% in all
samples. The structural features of the amorphous geopolymer, as resumed by the
broad hump of XRD patterns, are not modified by addition of RM. The physical properties of geopolymers are not significantly affected by RM, as all
samples exhibit high values of water absorption and low apparent density. The
chemical stability is good: sodium leaching test gave leachate concentrations close to
100 ppm without evidences of deterioration of mechanical performance. Red mud
influences the mechanical strength during curing (especially at the higher amounts of
RM but the lower additions, cured for 28 days showed good compressive strength). In general, incorporation of red mud, zeolite filler and diatomites tometakaolin
inmedium of alkali activators of low concentration provided formation of more eco-
friendly materials with high mechanical resistances and water treatment capabilities. Brunauer S, Emmett PH, Teller EJ (1938) Adsorption of gases on multimolecular layers. J Am
Chem Soc 60:309–319
Mackenzie K, Welter M (2014) Geopolymer (aluminosilicate) composites: synthesis, properties
and applications. Adv Ceram Matrix Compos. 445–470 2
Methods and Approaches New geopolymer formulations were designed by sodium silicate/NaOH/KOH activa-
tion of metakaolin, zeolites, diatomites and red mud mixtures. The effects of source
materials on the microstructure and mechanical properties were studied. Geopolymers were prepared using commercial metakaolin (1200S, AGS Mineraux,
France, D50 = 1.1 lm, bulk density = 296 g/dm−3), diatomite from Rio Maior and
Amieira deposits (Portugal), zeolite (ZeoBau micro 50, from Nižný Hrabovec, Zeocem,
Slovakia, CEC = 83 meq/100 g, SSA = 1663 m2/kg, particle size 0–0.05 mm, bulk
density = 500–600 g/dm−3, more information about Nižný Hrabovec deposit is
available on http://www.iza-online.org/natural/), red mud from aluminium metallurgies
wastes, hydrated sodium silicate (Merck, Germany Merck, Germany; 8.5 wt% Na2O,
28.5 wt% SiO2, 63 wt% H2O) and sodium and potassium hydroxide (ACS AR Ana-
lytical Reagent Grade Pellets). y
g
The role of the above ingredients in the preparation of geopolymers is as follows:
metakaolin was used as a precursor of aluminium; diatomite was used as a precursor of
silica; zeolite was used as filler with high specific surface area and cation exchange
capacity; red mud as precursor of aluminium and iron; sodium silicate was used as a
source of silicon and sodium and potassium hydroxide as alkaline activators for dis-
solution of aluminosilicate. Water was the reaction medium. The X-ray diffraction was conducted on Philips X’Pert diffractometer using CuKa
radiation at a speed of 0.02°/s. The X’Pert HighScore (PW3209) program was used to
analyze XRD peaks. The chemical composition (major elements) of materials was
analysed using PANalytical Axios X-ray fluorescence spectrometer. Loss on ignition
was determined by heating the samples in an electrical furnace at 1000 °C during 3 h. The microstructural characterization was carried out by scanning electron microscopy
(SEM – Hitachi, SU 70) and energy dispersive X-ray spectrometry (EDS – EDAX with
detector Bruker AXS, software: Quantax) operated at 3–30 kV. The specific surface area of geopolymers (heated at 200 °C during 6 h) was
determined by BET method (Brunauer, Emmett and Teller, nitrogen gas adsorption at
77 K) (Brunauer et al. 1938). Compressive strength tests were carried out on 3 probes
of individual geopolymer on (SHIMADZU: AG-IC 100 kN) equipment, by applying a
maximum force of 5 kN at the speed of 50 N/s according to the Standard EN 1015-11. Adsorption experiments were carried out in batch using nitrate solutions of Pb2+, Zn2+,
Cu2+, Cd2+ and Cr3+. References Brunauer S, Emmett PH, Teller EJ (1938) Adsorption of gases on multimolecular layers. J Am
Chem Soc 60:309–319
Mackenzie K, Welter M (2014) Geopolymer (aluminosilicate) composites: synthesis, properties
and applications. Adv Ceram Matrix Compos. 445–470 384 F. Rocha et al. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Features of Production of Fine Concretes
Based on Clinkerless Binders
of Alkaline Mixing M. Salamanova1,2(&), S.-A. Murtazaev1,2,3, A. Alashanov1,
and Z. Ismailova1 1 Grozny State Oil Technical University named after Academician
M.D. Millionshchikov, Grozny, Russia
Madina_salamanova@mail.ru
2 Complex Research Institute named after H.I. Ibragimov,
Russian Academy of Sciences, Grozny, Russia
3 Academy of Sciences of the Chechen Republic, Grozny, Russia Abstract. The paper showed relevance and potential for the development of
clinkerless alkaline activation, since at the present the production of currently
leading “constructional” binder, Portland cement, has been increasing, and
carbon dioxide released during cement production has a negative effect on the
ecological situation of separate countries and the whole world. The world
community has long been concerned about the problem of switching to clink-
erless binders and building composites to replace resource-intensive cement, at
least in those areas of construction that do not need its high technical and
functional properties. We gave formulations and properties of clinkerless alka-
line activation based on highly dispersed mineral components, effective com-
positions of fine concretes based on the use of the proposed clinkerless alkaline
activation cements were obtained. It was theoretically substantiated and prac-
tically proved that Brønsted acid sites on the surface of highly active powders
accelerated synthesizing silica gel, supported polymerization of silicon-oxygen
anions, enhanced ion exchange reactions and stabilized intergranular contact
formation. Keywords: Portland cement Clinkerless binders Mineral powders
Alkaline mixer Liquid glass High dispersion Keywords: Portland cement Clinkerless binders Mineral powders
Alkaline mixer Liquid glass High dispersion 2
Methods and Materials This paper presents results of researches on the formulation and study of the properties
of alkaline mixing binders using of sedimentary and magmatic rocks: quartz sand,
limestone, volcanic tuff and silicified marl. 1
Introduction The environmental problem, created by the cement industry, is associated with con-
sumption of large volumes of raw materials and release of huge amounts of carbon
dioxide and dust during production of mineral binders into the atmosphere. A promising direction for solving this problem is the use of alkaline activation binders,
which can be produced both from wastes of the fuel and energy industry, if present in
the region, and using highly dispersed aluminosilicate additives, which chemical
composition is characterized by an increased content of aluminum and silicon (Niki-
forov et al. 2011; Salamanova et al. 2015; Strokova et al. 2013). © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 385–388, 2019. https://doi.org/10.1007/978-3-030-22974-0_93 386 M. Salamanova et al. 3
Results and Discussion To prepare highly dispersed powders from the studied rocks, they were preliminarily
crushed in a jaw crusher, and then subjected to fine grinding for 1 h in a vibratory ball
mill. At the next stage, a number of Bronsted active crystallization centers on the surface
of the mineral powder was studied by the method (Strokova et al. 2013) of determining
the exchange capacity with respect to calcium ions (Table 1). Table 1. Surface activity of fine powders Table 1. Surface activity of fine powders
No. Mineral powder
Coefficient
of activity
Ka,%
Coefficient of
hydraulic
activity GHD
Number of active
centers of
crystallization,
mg eq/g
Specific
surface of
powders, m2/
kg
1
Quartz sand (QS) 22
1,76
21
810
2
Volcanic Tuff
(VT)
37
1,90
34
905
3
Limestone (L)
8
1,24
12
1060
4
Thermo-
activated marl
(TM) at 700 °C
62
2,03
42
1150
5
Sandstone (S)
10
1,44
16
1020 The coefficient of activity Ka,% The coefficient of hydraulic activity GAMD The
number of active centers of crystallization, mg eq/g Specific surface of powders, m2/kg. Thus, analyzing the results obtained we can state that the activity coefficients,
including the number of active crystallization centers, largely depend on the degree of
disclosure of defects formed during their grinding, in combination, all this contributes
to an increase in the reactivity of the powders used in concrete as highly dispersed
additives. After confirming the reactivity of the proposed powders, samples of 10 cm in size
were prepared using the mixture: a highly dispersed component, fractionated sand,
obtained by mixing in a ratio of 55:45% of screening crushing of rocks of the Argun
field and fine sand of the Chervlensky field. The mixing was carried out with liquid
glass, sodium hydroxide and accelerator of the precipitation of silica gel with sodium
fluoride in specified proportions. The prepared samples were hard in normal conditions
at a temperature of 20 ± 2 °C, but after 2 days the samples were placed in an oven at a Features of Production of Fine Concretes 387 temperature of 40–50 °C for several days. The test results of the investigated fine-
grained concretes based on alkaline activation binders are shown in Table 2. Table 2. 3
Results and Discussion Properties of fine concrete based on clinkerless binders of alkaline activation
No
compound
Materialconsumption per 1 m3
Density of concrete,
kg/m3
Strength at
compaction,
MPa at the
age, days
Fine
powders
Fractionated
sand
Na2SiO3
NaOH
Na2SiF6
7
28
1
QP - 480
1700
72,0
6,0
42,0
2240
11,8
24,7
2
TM - 480
1700
72,0
6,6
41,4
2250
31,4
40,5
3
VT - 480
1700
84,0
6,0
30,0
2246
25,3
34,2
4
L - 480
1700
60,0
3,6
56,4
2235
5,9
14,1
5
S - 480
1700
60,0
3,8
56,2
2239
6,6
15,9
Note: QP - quartz powder; TM - thermo-activated marl at 7000 °C; VT - volcanic tuff; L - limestone flour; S - sandstone; Na2SiO3 - sodium liquid
glass; Na2SiF6 - sodium silicofluoride; NaOH - sodium hydroxide. Note: QP - quartz powder; TM - thermo-activated marl at 7000 °C; VT - volcanic tuff; L - limestone flour; S - sandstone; Na2SiO3 - sodium liquid
glass; Na2SiF6 - sodium silicofluoride; NaOH - sodium hydroxide. The alkaline activation binder with the use of fine powders of thermally activated
marl showed the best results, so these particular samples of concrete were examined by
Quanta 3D scanning electron microscope (Fig. 1). Fig. 1. Micrographs of concrete on thermo-activated alkali activated marl Fig. 1. Micrographs of concrete on thermo-activated alkali activated marl In the contact area we determined a fairly strong accretion of particles of binder and
quartz sand, no defects on the surface in the form of growths or cracks, irregularities of
various shapes and sizes, and that individual particles have a needle-fibrous, vitreous
structure, which indicated an increased binder activity. High strength results of fine concrete using clinkerless alkaline mixing on the basis
of thermally activated marl are explained by formation of a durable geopolymer stone,
represented by a frame aluminosilicate shutter alkaline medium with the formation of a
3D aluminosilicate hydrogel (Murtazaev and Salamanova 2018; Soldatov et al. 2016). 388 M. Salamanova et al. 4
Conclusions Thus, we theoretically justified and practically proved that Brønsted acid sites on the
surface of highly active powders accelerated process of synthesizing silica gel, pro-
moted the polymerization of silicon-oxygen anions, enhanced ion exchange reactions
and stabilized intergranular contact formation. The results of the researches significantly expanded the field of application of
clinkerless binders on a liquid-glass binder and might enable partial replacement of
expensive and energy-intensive portland cement in the construction industry. Impact of Thermal Modification on Properties
of Basalt Fiber for Concrete Reinforcement
V. Strokova, V. Nelyubova(&), I. Zhernovsky, O. Masanin,
S. Usikov, and V. Babaev V. Strokova, V. Nelyubova(&), I. Zhernovsky, O. Masanin,
S. Usikov, and V. Babaev Department of Materials Science and Materials Technology,
Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
vvnelubova@gmail.com Abstract. The paper shows feasibility and efficacy of thermal modification of
basalt fiber to increase its corrosion resistance and durability in a cement matrix. The authors justify the mechanism of phase and structure transformation of the
fiber subsurface layer providing its increased physicochemical properties. Keywords: Basalt fiber Fiber Thermal modification Oxidation © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 389–392, 2019.
https://doi.org/10.1007/978-3-030-22974-0_94 References Murtazaev S-A, Salamanova M (2018) Prospects for the use of thermoactivated raw materials of
aluminosilicate nature. Volga Sci J 46(2):65–70 Nikiforov EA, Loganina VI, Simonov EE (2011) The influence of alkaline activation on the
structure and properties of diatomite. Bull BSTU Named After V.G. Shukhov (2):30–32 Salamanova MSh, Saidumov MS, Murtazaeva TS-A, Khubaev MS-M (2015) High-quality
modified concretes based on mineral additives and superplasticizers of various nature. Sci
Anal Mag “Innov Invest” (8):159–163 Soldatov AA, Sariev IV, Zharov MA, Abduraimova MA (2016) Building materials based on
liquid glass. In: The collection: actual problems of construction, transport, engineering and
technosphere safety materials of the IV-th annual scientific-practical conference of the North-
Caucasian Federal University. N.I. Stoyanov (executive editor), pp 192–195 Strokova VV, Zhernovskiy IV, Maksakov AV (2013) Express-method for determining the
activity of silica raw material for production, granulated nanostructuring aggregate. Constr
Mater (1):38–39 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches The main component in the work was basalt fiber produced at Novgorod glass fiber
plant using machine factory BASK by blowing the melt by vertical air stream. The method for investigation of thermal treatment impact on the fiber properties
included step-wise heating of the fiber from 300 to 700 °C (700 °C is the working
temperature of the basalt fiber, Twork) with the step of 100°. The isothermal holding
time was 30 min, which considered the microscopic dimensions of fibers and striving
to provide high productivity of the thermal treatment process. The fiber was cooled
down in air by convective heat exchange. The fiber specimens then were tested for
alkali resistance in model solutions represented by cement water extract. 1
Introduction Under the conditions of real operation, the elements of concrete structures suffer cyclic,
fatigue, impact, stretching and twisting loads, which causes uncontrolled cracking and
consequent destruction of the cement matrix (Lesovik et al. 2018). This problem was solved by using fibers to reinforce concrete matrix, which pre-
vents brittle fracture of concrete and enables control of cracking (Klyuyev et al. 2013). The analysis of approaches to enhancing the corrosion resistance of glass fiber in
alkaline medium demonstrates that the thermal treatment of basalt fiber without its
crystallization with initiation of a number of processes that increase alkali resistance
and strength of the fibers should be considered as the most advanced and economically
feasible method for increasing the stability of basalt fiber for its consequent application
for concrete production on the basis of cement binders. (Knot’ko et al. 2007; Knotko
et al. 2011) According to the accepted working hypothesis, the increase of alkali resistance of
fiber surface should be achieved by thermal treatment. This method is considered to be
the simplest, affordable and economically reasonable. Its technical performance is
predetermined by a range of physicochemical processes taking place in glass fibers
during thermal treatment: oxidation, structure and diffusive rearrangement of ions in the
material, annealing and densification of the structure, pre-crystallization and crystal-
lization, etc. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 389–392, 2019. https://doi.org/10.1007/978-3-030-22974-0_94 © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 389–392, 2019. https://doi.org/10.1007/978-3-030-22974-0_94 © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 389–392, 2019. https://doi.org/10.1007/978-3-030-22974-0_94 V. Strokova et al. 390 3
Results and Discussion According to preliminary data on the resistance of the fiber from different producers in
alkaline and acidic medium, the chosen basalt fiber stands out for its high alkali and
acid resistance. This is explained by the wash-out of cations persisting in vitrified phase
sue to alkaline hydrolysis into the silicon dioxide solution with formation of aluminates
and zincates with consequent liberation of anions SiO4
4−, Si2O5
2−and SiO3
2−. Insol-
uble complex aluminate and zincate salts accumulate on the chemically active fiber
surface, which inhibits further leaching of silicon. In the absence of further introduction
of alkaline component for supporting necessary pH, the decomposition retards. The corrosion of basalt fiber and its consequent disruption is caused by both
interaction with hydroxyl groups of silicon-oxygen radicals and capability of cations
dissolved in water alkaline medium to exchange with cations comprising the basalt
fiber. Noteworthy, during the utilization of basalt fiber in real conditions in concrete, the
fiber dissolution degree will be not that significant, since the solution processes will die
down along with curing and solidification of the cement. Nevertheless, the chemical
processes involving the fiber that take place in concrete during its life should not be
underestimated. This necessitates the development of the fiber modification method to
improve its resistance to aggressive alkali medium. The analysis of obtained data (Fig. 1) confirmed that thermal treatment of fiber at
500 °C was the most effective, as it promoted its alkali resistance as compared to non-
treated fiber by 35.3% (after 28 days). Further increase of the treatment temperature
was useless because, firstly, the fiber alkali resistance reduced; secondly, it was not
economically reasonable. The effect of increased alkali resistance at 300 and 400 °C
amounted to 26.7 and 30%, respectively. Heating of basalt fibers leads to negligible reduction of their mass due to loss of
adsorbed and chemically bound water, while further heating increases the mass by
binding of air oxygen during divalent iron FeO oxidation into trivalent Fe2O3. The
increased oxidation degree of iron cations leads to their decreased ion radius and
reduction of the coordination number with formation of groups of iron-oxygen tetra-
hedrons [FeO4]−Na+ that are embedded into the glass structure forming complex iron-
aluminum-silicon-oxygen framework and increasing the degree of association, stability Impact of Thermal Modification on Properties of Basalt Fiber 391 Fig. 1. Alkali resistance of fiber versus preliminary treatment temperature and testing time Fig. 1. 3
Results and Discussion Alkali resistance of fiber versus preliminary treatment temperature and testing time and density of the structure. Similar structural restructuring triggers the diffusion
redistribution of modifying cations (Na+, Ca2+) in the fiber. The thermal treatment is accompanied by microdefect healing, structure densifi-
cation and its approaching to stable equilibrium state. The processes described above are most active at 500 °C—the temperature com-
parable with the glass transition temperature Tg (corresponds to the viscosity of
1012,3 Pa s and transition of glass from solid into plastic state)—which conditions the
highest alkali resistance of fibers and stability of their work in cement concretes. 10
7 At higher temperatures (600–700 °C) and decreased viscosity (1010–107 Pa s), the
surface of basalt fibers suffers the manifestation of structural pre-crystallization and
crystallization processes accompanied by the formation of diverse defects, which
increase the free energy of the material and makes it more chemically active. This
results in accelerated interaction with alkali and increased fiber mass losses. Besides,
the thermal treatment at high temperature leads to the strength loss of basalt fibers: at
600 °C to negligible strength decrease; at 700 °C to the loss of 50% of initial strength. Thermal treatment of basalt fibers at 500 °C initiated a number of important
physicochemical and structural processes leading to the modification of glass fiber
surface with acquisition of higher chemical resistance. The holding close to the glass transition temperature Tg leads to gradual densifi-
cation of the structure and its transition into more stable equilibrium state with higher
chemical resistance. During the structure stabilization, numerous microdefects heal
inside and on the surface of the fiber, which results in its decreased free energy and
chemical activity. The structure densification also decreases the rate of numerous
diffusion processes, which are the basis of any chemical reactions with solid phases. Activation of iron ion oxidation during thermal treatment leads to substantial
restructuring of the surface layer of basalt fiber: modifying ions of Fe2+ after oxidation
to Fe3+ embed as iron-oxygen tetrahedrons [FeO4]5−into aluminum-silicon-oxygen
lattice of the glass, thus increasing the degree of its association and chemical resistance. Preliminary 30-min processing of the basalt fiber at 500 °C by 35% increases its
resistance in alkaline medium of curing cement and can be recommended as effective, V. Strokova et al. 392 simple and economically reasonable method for modifying basalt fiber for its effective
implementation as a micro-reinforcing component of fibrous concretes. 3
Results and Discussion simple and economically reasonable method for modifying basalt fiber for its effective
implementation as a micro-reinforcing component of fibrous concretes. 4
Conclusions Thus, the processes initiated by thermal treatment promote the corrosion resistance of
fibers in alkaline medium, which prolongs the corrosion resistance and increases the
efficacy of fiber application as a micro-reinforcing component for cement composites. Acknowledgements. The work has been fulfilled within the project Federal Target Program of
Research and Development on “Priority Development Fields of science and technology sector in
Russia for 2014–2020”, the unique project number is RFMEFI58317X0063. 1
Introduction Activation is a set of measures aimed at increasing the activity of cement by exposing it
to various mechanical methods. As a result of such exposure, physical and chemical
processes of different nature occur in the raw materials, leading to a change in the
characteristics of the final product (Kuznetsova and Sulimenko 1985). As a rule, during
mechanical activation, cement is crushed with an increase in its specific surface. In a jet mill, activation occurs due to processing of cement in turbulent swirling
flows (Korchakov 1986), leading not only to grinding, but also to transformation of the
shape of particles from angular to rounded. References Klyuyev SV, Klyuyev AV, Sopin DM, Netrebenko AV, Kazlitin SA (2013) Heavy loaded floors
based on fine-grained fiber concrete. Mag Civil Eng 38(3):7–14 not’ko AV, Garshev AV, Davydova IV, Putlyaev VI, Ivanov VK, Tret’yakov YuD (2007
Chemical processes during the heat treatment of basalt fibers. Prot Metals 43(7):694–700 Knotko AV, Pustovgar EA, Garshev AV, Putlyaev VI, Tret’yakov YD (2011) A protective
diffusion layer formed on surface of basaltic fiberglass during oxidizing. Prot Metals Phys
Chem Surf 47(5):658–661 Lesovik VS, Glagolev ES, Popov DY, Lesovik GA, Ageeva MS (2018) Textile-reinforced
concrete using composite binder based on new types of mineral raw materials. In: IOP
conference series: materials science and engineering, vol 327, no 3 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Activation of Cement in a Jet Mill S. Titov(&) and A. Kazakov S. Titov(&) and A. Kazakov S. Titov(&) and A. Kazakov Federal State Budgetary Educational Institution, Russian University of Transport
(MIIT), Moscow, Russia
titovs3094@yandex.ru Federal State Budgetary Educational Institution, Russian University of Transport
(MIIT), Moscow, Russia
titovs3094@yandex.ru Federal State Budgetary Educational Institution, Russian University of Transport
(MIIT), Moscow, Russia
titovs3094@yandex.ru Abstract. Mechanical treatment of cement in a jet mill leads to an increase in
the strength of cement-sand stone in compression and during bending. The effect
of increased activity is achieved by changing the shape of cement particles from
angular to rounded. Keywords: Cement Jet mill Cement activity Compressive strength
Bending strength Particle shape factor © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 393–394, 2019.
https://doi.org/10.1007/978-3-030-22974-0_95 2
Methods and Approaches The analysis of the shape of cement particles modified in a jet mill was performed using
the proposed shape criterion: ki ¼ ðS
P2Þi
ð1Þ ð1Þ where S – area of the particle in the plane of its cross section, P – length of the
perimeter in the cross section of the particle; i – order of the criterion (i =1/n…,1,…,n,
where n – counting numerals). To determine the degree of influence of the particle shape on the properties of
cement systems, standard samples were tested and the results obtained were processed
using mathematical statistics methods. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 393–394, 2019. https://doi.org/10.1007/978-3-030-22974-0_95 S. Titov and A. Kazakov 394 4
Conclusions The increase in compressive strength and bending of the samples based on cement
processed in a jet mill is explained by the change in the shape of its particles from
angular to rounded, which is associated with the characteristics of the vortex activation
method. Acknowledgements. The authors express their gratitude to the research supervisor, D.Sc. (Eng),
prof. Kondrashchenko V.I. 3
Results and Discussion It was found that processing in a jet type mill leads to an increase in dispersity of
cement from 5% to 0.8–1.2% on sieve No. 008 with a non-significant change in normal
density of the cement paste. Increasing the fineness of cement grinding causes, as a rule, an increase in mixing
water consumption to obtain a mixture of equal workability. But the experiment
showed the opposite – the water-cement ratio of the solution prepared on cement,
processed in a jet mill, not only did not increase, but also decreased. This is due to the
change in the shape of cement particles from angular to rounded, which was statisti-
cally justified using the proposed shape criterion ki (1). The results of the tests showed an increase in strength of the samples based on
portland cement processed in a jet mill for compression by 38–68% and by bending by
12–25% compared to non-activated cement. The Law of Similarity and Designing
High-Performance Composites A. Tolstoy(&), V. Lesovik, E. Glagolev, and L. Zagorodniuk Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
tad56@mail.ru Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
tad56@mail.ru Abstract. The increasing requirements for the quality of building products and
constructions condition the importance of developing new ways of controlling
the processes of shaping the structure of composite materials. This paper con-
siders the aspects of dealing with the tasks of designing high-performance
powder composites factoring in the law of similarity. Keywords: Law of similarity High-performance materials Powder concrete
Technogenic raw materials © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 395–398, 2019.
https://doi.org/10.1007/978-3-030-22974-0_96 References Korchakov VG (1986) Aerodynamics of flows in jet mills when grinding silicate materials, Ph.D. (Eng) thesis, Kharkov, 168 p
Kuznetsova TV, Sulimenko LM (1985) Mechanical activation of portland cement raw mixtures. Cement, no 4, pp 20–21 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction In our days, erecting special-purpose buildings and structures with complex design, as
well as the so-called unique buildings, calls for using high-performance powder con-
crete, which composition differs from the composition of traditional normal concrete in
the increased proportion of cement, higher fineness of grain, complex composition and
increased dispersity of aggregate (Bazhenov et al. 2007; Tolstoy et al. 2018a, b;
Lesovik et al. 2015). The role of each of the source materials, as well as that of the
mechanism of interaction between them, increases manifold, with fundamental con-
cepts of the law of similarity can explain the nature of the processes that occur. The theoretical foundation for designing high-quality composites is a new cross-
disciplinary scientific school: geonics (geomimetics), which employs the results of
studies of natural processes and rocks to create building materials of the future
(Lesovik 2014; Elistratkin and Kozhukhova 2018; Dmitrieva et al. 2018). This allowed
to develop a system for designing powder concrete using raw materials that were
specially prepared through geological processes, i.e. they are genetically activated
(Tolstoy et al. 2014). A separate school is further distinguished: crystal energy science, the science about
the modern approaches to estimating the quantitative indicators of the properties of
materials and explaining the processes of hydrate formation (Lesovik and Evtushenko
2002). The criteria of applicability of mineral components should be: abundance, acces-
sibility, cost, constant composition (Fig. 1). © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 395–398, 2019. https://doi.org/10.1007/978-3-030-22974-0_96 A. Tolstoy et al. 396 Powder concrete
Active binding component: thin
mineral powders possessing bind-
ing properties due to synergistic
similarity
Partially active component: ul-
trafine solid aggreagate (0.315-0.63
mm frac-tion) with active surface
Organic component: plasti-
cizing water-reducing poly-
mer materials
Genesis
Technogenesis
Superplasticizers
Hyperplasticizers
Fine-grained
cement clicker
Mineral materials
from natural rock
Technogenic min-
eral materials
Rapid strength gain, high strength, low porosi-
ty
Fig. 1. Types of raw materials for making powder concrete
96
A. Tolstoy et al. Partially active component: ul-
trafine solid aggreagate (0.315-0.63
mm frac-tion) with active surface Organic component: plasti-
cizing water-reducing poly-
mer materials Active binding component: thin
mineral powders possessing bind-
ing properties due to synergistic
similarity Fig. 1. Types of raw materials for making powder concrete Here it is necessary to use the “experience” of geological processes, for example,
high-performance siltstones that are similar to powder concrete in their strength and
other properties. 1
Introduction To create a strong and lasting composite of powder concrete, it is
necessary to ensure the reliable physical, mechanical and performance characteristics of
the material structure taking into account all energy parameters of all the substances
involved, as well as the compliance of other properties. This structure should be similar
to natural materials in terms of the main properties and genesis. 2
Methods and Approaches The experiments involved mineral admixtures containing the aluminate and carbonate
components, as well as off-the-shelf polymer ones: Melflux 2651, Melment, and finely
ground quartzite sandstone, a by-product rock from the Kursk Magnetic Anomaly
(KMA). 3
Results and Discussion At the moment there is no unified approach to the technique of producing high-
performance compositions. Some researchers address the issue from the perspective of
technological mechanics, other approaches consider the rheological properties of the
system, optimization of grain size composition, and the relation between the optimal
structure and extreme properties of components (the “law of gauge” by I.A. Rybiev
(Rybev 1999)). The proposed methods of controlling the formation of structure in curing systems
of powder mixes with technogenic components allow to achieve the composite strength
of 80–100 MPa (Table 1). The Law of Similarity and Designing High-Performance Composites The Law of Similarity and Designing High-Performance Composites 397 Table 1. Comparative figures of normal concrete and powder concrete
Parameter
Values
Normal concrete Powder concrete
Average density, kg/m3
2200–2500
2300
Compressive strength, MPa
10–50
97.5
Water retention capacity, %
78–80
90
Strength-density ratio
0.17
0.36
Water resistance grade. W
2–4
4
Freeze-thaw resistance grade, F
50–150
300
Wear capacity, kg/m2
0.7–0.8
0.36
Shrinkage
No fractures
Thermal conductivity ratio, W/(m K) 0.8–1.2
1.29 A feasible foundation can be represented by the method of estimating the com-
position of high-density concrete taking into account the law of similarity of properties
(Tolstoy et al. 2018a, b). Dense structure of powder concrete is characterized by a virtually complete absence
of pores and microfractures. It was possible to implement this by using the adjusted
composition of the curing mass, introducing the necessary number of fine-grained
technogenic components, their densest packing, and the self-compacting effect of
curing. Study of the micrographs of the curing compositions revealed the following: – the microstructure of the cement rock obtained through intergrinding possesses
higher homogeneity than that obtained through separate grinding of components; – growth of needle-shaped crystals that permeate the volume of the material’s
structure was detected; – dense neogeneses are present near the aggregate grains; References Bazhenov IM, Demianova VS, Kalashnikov VI (2007) Modifitcirovannye vysokoprochnye
betony [Modified high-performance concretes], ASV, Moscow, 368 p Dmitrieva TV, Strokova VV, Bezrodnykh AA (2018) Influence of the genetic features of soils on
the properties of soil-concretes on their basis. Constr Mater Prod 1(1):69–77 Elistratkin MYu, Kozhukhova MI (2018) Analysis of the factors of increasing the strength of the
non-autoclave aerated concrete. Constr Mater Prod 1(1):59–68 Elistratkin MYu, Kozhukhova MI (2018) Analysis of the factors of
non-autoclave aerated concrete. Constr Mater Prod 1(1):59–68 Lesovik VS (2014) Geonika (Geomimetika) [Geonics (Geomimetics)]. Primery realizatcii v
stroitelnom materialovedenii [Examples of application in building materials science]. Bulletin
of BSTU named after V.G. Shoukhov, Belgorod, 206 p Lesovik VS, Zagorodniuk LK, Chulkova IL, Tolstoy AD, Volodchenko AA (2015) Srodstvo
struktur, kak teoreticheskaia osnova proektirovaniia kompozitov budushchego [Similarity of
structures as a theoretical foundation of designing the composites of the future]. Stroitelnye
materialy [Building materials], no 9, pp 18–22 Lesovik BC, Evtushenko EI (2002) Stabilizatciia svoistv stroitelnykh materialov na osnove
tekhnogennogo syria [Stabilizing the properties of building materials on the basis of
technogenic raw materials]. Izvestiia VUZov [Bul. HEIs]. 12:40–44 Rybev IA (1999) Otkrytie zakona stvora i vzaimosviaz ego s zakonom kongruentcii v stroitelnom
materialovedenii [Discovering the gauging law and its relation to the law of congruence in
building materials science]. Stroitelnye materialy [Build Mater] 12:30–31 Tolstoy AD, Lesovik VS, Milkina AS (2018a) Osobennosti struktury betonov novogo pokoleniia
s primeneniem tekhnogennykh materialov [Specifics of the structure of new-generation
concretes using technogenic materials]. Russ Automob Highw Ind J Sect III Constr Arch. 15(4, 62):588–595 Tolstoy AD, Lesovik VS, Kovaleva IA (2014) Organomineralnye vysokoprochnye dekorativnye
kompozitcii [Organic and mineral high-performance decorative compositions]. Bull BSTU
Named After V.G. Shoukhov 5:67–69 Tolstoy AD, Lesovik VS, Glagolev ES, Krymova AI (2018b) Synergetics of hardening
construction systems. In: IOP conference series: materials science and engineering, vol 327,
p 032056. https://doi.org/10.1088/1757-899x/327/3/032056 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. 4
Conclusions This way, introducing the theory of geonics (geomimetics), particularly the law of
similarity, allows to obtain high-performance powder concretes, all components of
which possess a high adhesion and similar deformation and thermal characteristics. The
first attempt to test this law allowed to obtain composites with the maximum com-
pressive strength of up to 100 MPa. Acknowledgements. The work has been fulfilled within the project Federal Target Program of
Research and Development on “Priority Development Fields of science and technology sector in
Russia for 2014–2020”, unique project number is RFMEFI58317X0063. A. Tolstoy et al. 398 References If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Genesis of Clay Rock of the Incomplete Stage
of Mineral Formation as a Raw Material Base
for Autoclaved Materials A. Volodchenko(&) and V. Strokova Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
volodchenko@intbel.ru Abstract. We illustrated possible application of clay rock of incomplete stage
of mineral formation to produce autoclaved silica materials. A distinguishing
feature of this rock was the presence of thermodynamically unstable com-
pounds. Weathering processes resulted in partial disintegration of the rock,
formation of defects in the crystalline lattice of the rock-forming minerals, and
partial amorphization of both silicates and aluminosilicates, which reduced the
energy potential of the raw materials. Thus, it was possible to use hydrothermal
conditions to boost the formation of neogeneses of cementing compounds and
reduce power consumption of the production of autoclaved materials. Keywords: Lime Clay rock Autoclaved silica Keywords: Lime Clay rock Autoclaved silica © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 399–402, 2019.
https://doi.org/10.1007/978-3-030-22974-0_97 2
Results and Discussion Some of the most proliferated rocks are deposits of clay. They are formed through
weathering of igneous and metamorphic rocks. The process of weathering of felsic rock passes the following stages: physical
weathering (clastic stage), start of the chemical weathering with illite formation (siallite
stage), chemical weathering with kaolinite formation (acidic siallite stage), and com-
plete decomposition of silicates with the formation of aluminum, ferrous, and silicon
oxides (allite stage). Weathering of rock results in the formation of minerals pre-
dominantly of the montmorillonite group. The processes of weathering can be represented as the destruction of the crystalline
structure of the source minerals followed by a transfer through the pseudo-amorphous
state and then the formation of the crystalline structure of new rock-forming minerals. However, the ideal amorphous (non-crystalline) state, as well as crystalline state, is not
achieved and the transition through it is considered conditional. This results in the
change of the source framework structure of the feldspar into the layered structure of
clay minerals. This transition results in rocks that can be characterized as products of the inter-
mediary stage of weathering that occupy a significant stretch in the line of sub-stance
transformation and predominate in nature. The rock-forming minerals of these for-
mations include the X-ray amorphous phase, illites, mixed-layer minerals, and
imperfectly structured kaolinite and montmorillonite. The formation of these com-
pounds is accompanied by disordering of the crystalline structure of the source min-
erals, which increases the thermodynamic instability of the rock. The rocks in the intermediary stage of weathering that were mechanically and
chemically activated by exogenous geologic processes, i.e. those that belong to the
starting stage of chemical weathering that results in the formation of defects in the
structure of silicates and aluminosilicates and the formation of illites, are virtually never
used. However, due to their thermodynamic instability, these rocks are the most effi-
cient raw material as components of the systems that must be highly reactive. In
particular, such rocks can be used as components of hydro-thermal curing binders,
which allows to manage the processes of the formation of the structure of new-
generation autoclave materials. 1
Introduction The traditional technology of manufacturing of silica materials is based on using lime
and silica sand as raw materials, and the reserves of the latter are getting depleted. Furthermore, this technology is marked with a high power consumption. The con-
sumption of power for grinding the binder and hydrothermal treatment can be reduced
by using highly reactive polymineral raw materials, which would allow not only to
replace silica sand but also intensify the technological processes. A prospective raw
material for producing autoclaved silica materials is the clay rock of the incomplete
stage of mineral formation, which is currently unconventional in the building industry
(Lesovik 2012; Lesovik et al. 2014; Alfimova and Shapovalov 2013; Volodchenko
et al. 2015; Volodchenko et al. 2016; Alfimov et al. 2006; Alfimova et al. 2013). As
compared with the traditional silica sand, this rock ensures the synthesis of neogeneses
in a more complex system “CaO–[SiO2–Al2O3–(MgO)]–H2O”, which the reduction of
power consumption of manufacturing and improving physical, mathematical, and
performance properties of both solid and cellular autoclaved materials (Volodchenko
and Lesovik 2008a, 2008b; Volodchenko 2011). The purpose of this paper is to study the genesis of clay rock of the incomplete
stage of mineral formation as raw material for producing autoclaved silica materials. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 399–402, 2019. https://doi.org/10.1007/978-3-030-22974-0_97 400 A. Volodchenko and V. Strokova 2
Results and Discussion Based on the data about the genesis of the weathering rind rocks, the processes of
phase formation in artificial systems of autoclaved curing and how they compare with
their natural counterparts of mineral formation, a diagram of the exogenous processes
of the genesis of clay rocks as a raw material base for autoclaved materials is proposed
(Fig. 1). The rocks that formed after the second (siallite stage) and partially the third (acidic
siallite) stage of weathering belong to the rocks of the incomplete stage of mineral
formation that contain compounds of the intermediary stage of weathering: the source
minerals have decomposed while the final ones have not formed yet. Such minerals are
characterized by the presence of crystalline structure defects and, consequently, ther-
modynamic instability. This kind of rock can either remain at the place of its formation Genesis of Clay Rock of the Incomplete Stage of Mineral Formation 401 Bio- and chemical
decomposition (sial-
lite stage)
Mechanical
destruction
Sedimentation
Diagenesis
Igneous
rocks
Metamorphic
rocks
Weathering
Acidic
Siallite stage
Sedimentary rocks
Raw materials
for ceramic materials,
cement, etc. Allite stage
Bauxites,
laterites
Secondary clay rocks
of the incomplete
stage
of mineral formation
Primary clay
rocks of the incom-
plete stage of miner-
al formation
Diagenesis
Redeposition
Mechanically and chemi-
cally activated rocks
Raw materials
for autoclaved
materials
Fig. 1. Diagram of the exogenous processes of the genesis of clay rocks as raw materials base
for autoclaved materials Fig. 1. Diagram of the exogenous processes of the genesis of clay rocks as raw materials base
for autoclaved materials or redeposit in new places as the result of transfer and diagenesis. These rocks are
useful as raw materials for autoclaved curing materials. Thus, weathering of source rocks of different compositions leads to the formation
of mineral resources that are different in their composition and industrial significance. If
one considers the weathering rind profile, it can be seen that at the moment the building
materials industry is using the lower horizon (source rocks) and products of the final
stage of weathering: kaolinite and montmorillonite clays as well as bauxites. Clay rocks of the incomplete stage of mineral formation are some of the most
inconsistent in terms of composition and the structure of the sedimentation mass
deposits. 2
Results and Discussion In terms of the crystallochemical features of the rock-forming minerals, it is
necessary to note the high content of the crystalline structure defects, the dis-order of
the interlayer aluminosilicate stacks in the layered structure of minerals, etc. The
crystalline lattice of the typical clay minerals that belong to layered silicates is char-
acterized by the alteration of silicon-oxygen tetrahedra and hydroxyl octahedra. To
some degree, clay minerals can be considered objects, where the nanoparticles (ele-
mentary layer packets) are packed into the microscopic structures, where the properties
of the individual constituent particles are concealed due to a strong interparticle
interaction. 3
Conclusions Thus, lay rocks of the incomplete stage of mineral formation are energy-saturated
deposits. Due to the composition and crystallochemical features of the rock-forming
minerals, these rocks can be recommended as energy-efficient raw materials for the
production of autoclaved curing materials. A distinguishing feature of this rock is the
presence of thermodynamically unstable compounds. At the same time, aluminosili-
cates are characterized by variable chemical composition and the imperfection of their
crystalline lattice. Weathering processes result in partial disintegration of rock, for-
mation of defects in the crystalline lattice of the rock-forming minerals, and partial
amorphization of both silicates and alumino-silicates, which reduces the energy A. Volodchenko and V. Strokova 402 potential of the rock-forming minerals. Thus, it is possible to use hydrothermal con-
ditions to boost the formation of neogeneses of cementing compounds and reduce the
power consumption of the production of autoclaved materials. potential of the rock-forming minerals. Thus, it is possible to use hydrothermal con-
ditions to boost the formation of neogeneses of cementing compounds and reduce the
power consumption of the production of autoclaved materials. Acknowledgements. The work is realized in the framework of the Program of flagship uni-
versity development on the base of the Belgorod State Technological University named after
V.G. Shukhov, using equipment of High Technology Center at BSTU named after
V.G. Shukhov. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 403–406, 2019.
https://doi.org/10.1007/978-3-030-22974-0_98 1
Introduction The properties of any composite materials directly depend on the characteristics of their
base matrix. This also fully applies to composites based on binding agents with various
composition, which quality indicators are set as a result of system hardening (Strokova
et al. 2015; Kozhukhova et al. 2016; Nelyubova et al. 2017; Dmitrieva et al. 2018;
Shulpekov et al. 2018). In this regard the study of phase formation in such systems
seems quite relevant. Construction materials based on aluminate cements have different properties than
those based on portland cement. In particular, they are characterized by a high thermal
stability and resistance to acid corrosion (Kuznetsova 1986; Kuznetsova et al. 1989). The calcium aluminate hydrates, which are formed through hardening of aluminate
h f ll
i
h
CAH
C AH
C AH
AH ( ibb i ) F Construction materials based on aluminate cements have different properties than
those based on portland cement. In particular, they are characterized by a high thermal
stability and resistance to acid corrosion (Kuznetsova 1986; Kuznetsova et al. 1989). Construction materials based on aluminate cements have different properties than
those based on portland cement. In particular, they are characterized by a high thermal
stability and resistance to acid corrosion (Kuznetsova 1986; Kuznetsova et al. 1989). The calcium aluminate hydrates, which are formed through hardening of aluminate
cement, represent the following phases: CAH10, C2AH8, C3AH6, AH3 (gibbsite). From
this point onward the ‘cement’ notation in formulas of chemical compounds will be
used (C – CaO, A – Al2O3, H – OH). The calcium aluminate hydrates, which are formed through hardening of aluminate
cement, represent the following phases: CAH10, C2AH8, C3AH6, AH3 (gibbsite). From
this point onward the ‘cement’ notation in formulas of chemical compounds will be
used (C – CaO, A – Al2O3, H – OH). The hydrate mineral formation in aluminate cements is characterized by the
dependence of C-A-H phases on temperature. At a temperature below 15 °C the main
reaction-active CA phase is hydrated according to the following scheme: CA + 10H ! CAH10. At room temperature the C-A-H is formed according to the following scheme:
2CA + 11H!C2AH8 + AH3. At higher temperatures (above 28 °C) the metastable
hydrates CAH10 and C2AH8 will transform into stable hydrate C3AH6 according to the
following equations: 3CAH10!C3AH6 + 2AH3 + 18H and 3C2AH8!2C3AH6 +
AH3 + 9H (Rashid et al. 1994). Abnormal Mineral Formation in Aluminate
Cement Stone Abnormal Mineral Formation in Aluminate
Cement Stone
I. Zhernovsky(&), V. Strokova, V. Nelyubova, Yu. Ogurtsova,
and M. Rykunova
Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
zhernovsky.igor@mail.ru, zhernovskiy.iv@bstu.ru I. Zhernovsky(&), V. Strokova, V. Nelyubova, Yu. Ogurtsova,
and M. Rykunova
Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
zhernovsky.igor@mail.ru, zhernovskiy.iv@bstu.ru I. Zhernovsky(&), V. Strokova, V. Nelyubova, Yu. Ogurtsova,
and M. Rykunova Abstract. The paper describes the results of the study on the correlation between
the strength properties and the concentration of calcium aluminate hydrates during
long hardening of aluminate cement in aqueous medium. It shows the abnormal
nature of hydrate mineral formation in the system of hardening aluminate cement. Keywords: Aluminate cement Calcium aluminate hydrates References Alfimov SI, Zhukov RV, Volodchenko AN, Yurchuk DV (2006) Technogenic raw materials for
silicate hydration hardening. Mod High Technol 2:59–60 Alfimova NI, Shapovalov NN (2013) Materials autoclaved using man-made aluminosilicate
materials. Fundam Res 6(3):525–529 Alfimova NI, Shapovalov NN, Abrosimova OS (2013) Operational characteristics of silica brick,
manufactured using man-made aluminosilicate materials. Bulletin of BSTU named after V.G. Shukhov, vol 3, pp 11–14 Lesovik VS (2012) Geonik. Subject and tasks. Publisher Belgorod State Technological
University. VG Shukhov, p 219 Lesovik VS, Volodchenko AA, Svinarev AA, Kalashnikov NV, Rjapuhin NV (2014) Reducing. World Appl Sci J 31(9):1601–1606 Volodchenko
AA,
Lesovik
VS,
Volodchenko
AN,
Glagolev
ES,
Zagorodnjuk
LH,
Pukharenko YV (2016) Int J Pharm Technol 8(3):18856–18867 Volodchenko AA, Lesovik VS, Volodchenko AN, Zagorodnjuk LH (2015) Int J Appl Eng Res
10(24):45142–421149 Volodchenko AN, Lesovik VS (2008a) Increasing production efficiency of autoclave materials. In: Proceedings of the higher educational institutions. Building, vol 9, pp 10–16 Volodchenko AN, Lesovik VS (2008a) Increasing production efficiency of autoclave materials. Volodchenko AN, Lesovik VS (2008a) Increasing production efficiency of autoclave materia
I
P
di
f th hi h
d
ti
l i
tit ti
B ildi
l 9
10 16 ,
(
)
g p
y
In: Proceedings of the higher educational institutions. Building, vol 9, pp 10–16 Volodchenko AN, Lesovik VS (2008b) Autoclave silicate materials with nanometer-sized
materials. Build Mater 11:42–44 Volodchenko AN (2011) Features magnesia clay interaction with the calcium hydroxide in the
synthesis and the formation of tumors microstructure. Bulletin of Belgorod State Techno-
logical University. VG Shukhov, vol 2, pp 51–55 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Materials and Methods The aluminate cement GTS-50 produced by JSC Pashiysky Cement and Metallurgical
Plant (Perm Region, Russian Federation) was used in the study as a binding agent. Cubes with a 2 cm side are formed from cement paste (water-cement ratio equal 0.3). After hardening on the 1st day the samples were taken from a mold and placed into the
desiccator with 100% air humidity. After 28 days the samples were dried in a drying
cabinet at 80 °C and placed in water, from which they were taken for further study in 1,
2, 3 and 4 months. The study methods included the compression strength test on PGM-100 MG4 press
(average against three measurements) and the quantitative full-scale XRF to define
potential changes in mineral composition. The diffraction spectra of samples were obtained via the ARL X’TRA diffrac-
tometer (kCu) in the range of diffraction angles 2hº = 4-64, step angle – 0.02º. The
diffraction spectra were smoothened prior to treatment. The quantitative full-scale XRF was carried out via the DDM v.1.95e software for a
difference curve derivative (Solovyov 2004). I. Zhernovsky et al. The paper provides the results of hydration hardening of aluminate cement with
long-term exposure to aqueous medium, which contradict the above ideas. 1
Introduction Thus, according to insights into hydration of aluminate cements, the hydrate phases
during long hydration shall be presented as follows: C3AH6 + AH3. 404 I. Zhernovsky et al. Abnormal Mineral Formation in Aluminate Cement Stone 405 Abnormal Mineral Formation in Aluminate Cement Stone Table 1. Concentration of hydrate phases (wt. %)
Time in aqueous medium, months CAH10 C2AH8 C3AH6 AH3
0
18.8
3.5
3.1
2.0
1
26.5
4.4
8.2
5.9
2
9.0
2.1
6.8
25.1
3
33.5
3.5
4.7
6.3
4
37.2
5.8
5.6
3.0
Time in water, months
Compression strength,
MPa (1)
САН10 concentration,
wt. % (2)
Fig. 1. Compression strength of aluminate cement stone and CAH10 concentration depending
on time
Abnormal Mineral Formation in Aluminate Cement Stone
405 Table 1. Concentration of hydrate phases (wt. %) Time in aqueous medium, months CAH10 C2AH8 C3AH6 AH3
0
18.8
3.5
3.1
2.0
1
26.5
4.4
8.2
5.9
2
9.0
2.1
6.8
25.1
3
33.5
3.5
4.7
6.3
4
37.2
5.8
5.6
3.0 Fig. 1. Compression strength of aluminate cement stone and CAH10 concentration depending
on time 4
Conclusions The paper describes the results of the study on the correlation between the strength
properties and the concentration of calcium aluminate hydrates during long hardening
of aluminate cement in aqueous medium. At present it is impossible to give unam-
biguous interpretation of the observed abnormal compositional changes of hydrate
phases of hydrated aluminate cement during long hardening. Acknowledgements. The study is carried out in the framework of the State Task of the RF
Ministry of Education and Science No. 7.872.2017/4.6. Development of principles for the design
of ecologically positive composite materials with prolonged bioresistance 2017–2019. 3
Results and Discussion According to XRF, the mineral composition of GTS-50 cement is presented by the
following crystal formations (wt.%): CA (28.3), CA2 (9.2), C12A7 (1.4), akermanite-
gelenite (15.6), b-C2S (10.0), a’H-C2S (10.2), wollastonite 2M (7.6), dolomite (10.6),
perovskite (7.2). According to XRF, the mineral composition of GTS-50 cement is presented by the
following crystal formations (wt.%): CA (28.3), CA2 (9.2), C12A7 (1.4), akermanite-
gelenite (15.6), b-C2S (10.0), a’H-C2S (10.2), wollastonite 2M (7.6), dolomite (10.6),
perovskite (7.2). X-ray diagnostics of mineral phases in hydrated samples of aluminate cement
indicated the presence of CAH10, C2AH8, C3AH6, AH3, akermanite-gelenite, b-C2S,
a’H-C2S, wollastonite 2M, dolomite and perovskite. Due to lack of C2AH8 structural model, the quantitative XRF was combined with
the approach suggested by Cuberos et al. (2009). As data show (Table 1), for nearly 5 months (28 days in damp atmosphere and 4
months in water) CAH10 remains the main hydrate phase. The reduction of CAH10
concentration on the 2nd month in water, which correlates well with the increase of
crystal gibbsite (AH3), does not lead to the similar increase of C3AH6. Most likely it is
caused by the formation of C3AH6 in its cryptocrystalline state. The abnormal nature of hydrate mineral formation includes the increase of CAH10
metastable hydrate concentration starting from the 3rd month. At the same time the
temporal change of C3AH6 concentration coincides well with the change of com-
pression strength of the studied samples (Fig. 1). Cuberos AJM, De la Torre ÁG, Martín-Sedeño MC, Moreno-Real L, Merlini M, Ornez LM,
Aranda MAG (2009) Phase development in conventional and active belite cement pastes by
Rietveld analysis and chemical constraints. Cem Concr Res 39:833–842
Dmitrieva TV, Strokova VV, Bezrodnykh AA (2018) Influence of the genetic features of soils on
the properties of soil-concretes on their basis. Constr Mater Prod 1(1):69–77 References Cuberos AJM, De la Torre ÁG, Martín-Sedeño MC, Moreno-Real L, Merlini M, Ornez LM,
Aranda MAG (2009) Phase development in conventional and active belite cement pastes by
Rietveld analysis and chemical constraints. Cem Concr Res 39:833–842
Dmitrieva TV, Strokova VV, Bezrodnykh AA (2018) Influence of the genetic features of soils on
the properties of soil-concretes on their basis. Constr Mater Prod 1(1):69–77 Cuberos AJM, De la Torre ÁG, Martín-Sedeño MC, Moreno-Real L, Merlini M, Ornez LM,
Aranda MAG (2009) Phase development in conventional and active belite cement pastes by
Rietveld analysis and chemical constraints. Cem Concr Res 39:833–842 Dmitrieva TV, Strokova VV, Bezrodnykh AA (2018) Influence of the genetic features of soils on
the properties of soil-concretes on their basis. Constr Mater Prod 1(1):69–77 I. Zhernovsky et al. 406 Kozhukhova NI, Chizhov RV, Zhernovsky IV, Strokova VV (2016) Structure formation of
geopolymer perlite binder vs. type of alkali activating agent. ARPN J Eng Appl Sci 11
(20):12275–12281 Kuznetsova TV (1986) Aluminate and sulfo-aluminate cements, Moscow Kuznetsova TV, Kudryashov IV, Timashev VV (1989) Physical chemistry of binding materials,
Moscow Nelyubova V, Pavlenko N, Netsvet D (2015) Cellular composites with ambient and autoclaved
type of hardening with application of nanostructured binder. In: IOP Conference series:
materials science and engineering, vol 96, no 1, p 012010 Nelyubova VV, Strokova VV, Sumin AV, Jernovskiy IV (2017) The structure formation of the
cellular concrete with nanostructured modifier. Key Eng Mater 729:99–103 Rashid S, Barnes P, Bensted J, Turrillas X (1994) Conversion of calcium aluminate cement
hydrates re-examined with synchrotron energy-dispersive diffraction. J Mater Sci Lett
13:1232–1234 Shulpekov AM, Lepakova OK, Radishevskaya NI (2018) Phase – and structural formation in the
TIO2-AL-C system in the SHS process. Chem Bull 1(1):4–11 Solovyov LA (2004) Full-profile refinement by derivative difference minimization. J Appl
Crystallogr 37:743–749 Strokova VV, Botsman LN, Ogurtsova YN (2015) Impact of epicrystallization modifying on
characteristics of cement rock and concrete. Int J Appl Eng Res 10(24):45169–45175 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. References The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction The mechanoactivation dispergation of quartz materials is a widespread method of
technological processing of this mineral raw material in various fields of technological
mineralogy. In practice of technical petrogenesis of cast stone, forming the basis of synthesis of
inorganic silica-containing binding agents (Dmitrieva et al. 2018), the mechanoacti-
vation dispergation of quartz raw material holds a special place. The result of almost a century-long study of phase and structural transformations of
low-temperature quartz during mechanoactivation is the amorphicity of a surface layer
of quartz particles and the formation of nanosized b-quartz crystals in a-quartz matrix
(Zhernovsky et al. 2018). At the same time there was no study on structural trans-
formations of a-quartz within a quartz particle matrix during mechanoactivation. The
only exception is the study by Archipenko et al. (1987, 1990) concerning phase
transformations in mechanoactivated a-quartz. The task of this study is to fill the gap in
this matter partially. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 407–410, 2019.
https://doi.org/10.1007/978-3-030-22974-0_99 I. Zhernovsky(&) and V. Strokova I. Zhernovsky(&) and V. Strokova Belgorod State Technological University named after V.G. Shukhov,
Belgorod, Russia
zhernovsky.igor@mail.ru Abstract. The paper presents the study of mechanoactivation impact on crystal
structure of a-quartz. The volume of silicon-oxygen tetrahedron SiO4
4– is
accepted as the structural parameter depending on the mechanoactivation
degree. The paper compares the dependence of this parameter on temperature,
pressure and time of mechanoactivation of a-quartz in a planetary mill. Keywords: Quartz Crystal structure Silicon-oxygen tetrahedron
Mechanoactivation Keywords: Quartz Crystal structure Silicon-oxygen tetrahedron
Mechanoactivation 2
Materials and Methods The grinding of hydrothermal quartz (Ural) was carried out in the PULVERISETTE 6
classic line planetary mill (Fritsch, Germany) with lining and grinding bodies of
tungsten carbide. The milling time made 3, 12, 30, 60, 120, 180, 240, 300 and 360 min. 408 I. Zhernovsky and V. Strokova The diffraction spectra of samples are obtained using ARL 9900 Workstation
(kCo). Shooting interval – 2hº:8–80, step angle – 0.02. The specification of structural
parameters was carried out in DDM v.1.95e software for the difference curve derivative
(Solovyov 2004). 174-ICSD data (P3221) were used as a structural model. Profile
function – pseudo-Voigt. The specification of the profile parameters was carried out in
the anisotropic approximation. Thermal corrections were specified in anisotropic
option. Table 1 shows the experimental results. Table 1. Coordinates of a-quartz atoms after mechanoactivation Table 1. Coordinates of a-quartz atoms after mechanoactivation
Milling time, min
3
12
30
60
120
180
240
300
360
Si* x/a 0.4499 0.463
0.460
0.466
0.465
0.466
0.468
0.469
0.470
O x/a
0.403
0.416
0.413
0.419
0.418
0.419
0.422
0.422
0.423
O y/b
0.2660 0.2702 0.2697 0.2715 0.2668 0.272
0.2697 0.2721 0.2707
O z/c
0.7950 0.7921 0.7919 0.7905 0.7886 0.7868 0.7866 0.7887 0.7876
Note: Si y/a = 0, Si z/c = 2/3. Structural Transformations of Low-Temperature Quartz During Mechanoactivation 409 a
b
tetrahedrons
tetrahedrons
Temperature, ◦C
Pressure, kBar
Fig. 1. Calculated dependences of volumes of a-quartz silicon-oxygen tetrahedrons on thermal
(a) and baric (b) influences. (b) indicates the following data: ♦– according to Levien et al. (1980),
▲– according to Hazen et al. (1989), ■– according to Glinnemann et al. (1992). a
tetrahedrons
Temperature, ◦C b
tetrahedrons
Pressure, kBar Fig. 1. Calculated dependences of volumes of a-quartz silicon-oxygen tetrahedrons on thermal
(a) and baric (b) influences. (b) indicates the following data: ♦– according to Levien et al. (1980),
▲– according to Hazen et al. (1989), ■– according to Glinnemann et al. (1992). tetrahedrons
Milling time, min
Fig. 2. Dependence of volumes of a-quartz silicon-oxygen tetrahedrons on mechanoactivation
time Fig. 2. Dependence of volumes of a-quartz silicon-oxygen tetrahedrons on mechanoactivation
time 3
Results and Discussion The volume of silicon-oxygen tetrahedrons was chosen as a structural and sensitive
quartz parameter on mechanoactivation influence since it depended on several ele-
mentary structural parameters: bond length Si-O, bond angle O-Si-O and twist angle of
tetrahedrons (Goryaynov and Ovsyuk 1999). It is known that mechanoactivation is the result of two processes influencing the
material – local thermal influence and impact pressure. Hence, it is advisable to con-
sider structural changes of quartz during mechanoactivation in comparison with
changes during thermal and baric impacts. The following were used as references on
changes of quartz structural parameters: at a thermal influence – the work of Kihara
(1990), at a baric impact – works of Levien et al. (1980), Hazen et al. (1989) and
Glinnemann et al. (1992). The given results show that mechanoactivation alongside with thermal and baric
influences transforms the structure of a-quartz by reducing the volume of SiO4-tetra-
hedrons. At the same time the dependences of volumes of a-quartz silicon-oxygen
tetrahedrons on severity of exposure differ a lot (Figs. 1 and 2). The energy accumulation by quartz during mechanoactivation, as well as under
thermal and baric influence most likely happens due to the reduction of Si-O bond
length. The energy accumulated by quartz during mechanoactivation can be estimated by
comparing the dependences (Figs. 1a and 2). This comparison confirms that material
energy saturation after three-hour mechanoactivation is equivalent to its heating up to
500 °C. Structural Transformations of Low-Temperature Quartz During Mechanoactivation References Archipenko DK, Bokyi GB, Grigorieva TN, Koroleva SM, Yusupov TS (1987) On a new quartz
phase stable at room temperature and found during tribo-processing (via x-ray diffraction). Reports of the USSR Academy of Sciences, vol 296, pp 1370–1374 of the USSR Academy of Sciences, vol 296, p Archipenko DK, Bokyi GB, Grigorieva TN, Koroleva SM, Yusupov TS, Shebanin AP (1990)
Deformed quartz structures obtained after mechanoactivation. Reports of the USSR Academy
of Sciences, vol 310, pp 874–877 Dmitrieva TV, Strokova VV, Bezrodnykh AA (2018) Influence of the genetic features of soils on
the properties of soil-concretes on their basis. Constr Mater Prod 1:69–77 Glinnemann J, King HE, Schulz H, Hahn T, La Placa SJ, Dacol F (1992) Crystal structures of the
low-temperature quartz-type phases of SiO2 and GeO2 at elevated pressure. Zeitschrift fur
Kristallographie 198:177–212 g p
Goryaynov SV, Ovsyuk NN (1999) Twisting of a-quartz tetrahedrons at pressure close to
transition to an amorphous state. J Exp Theor Phys 69:431–435 Hazen RM, Finger LW, Hemley RJ, Mao HK (1989) High-pressure crystal chemistry and
amorphization of alpha-quartz. Solid State Commun 72:507–511 Kihara K (1990) An X-ray study of the temperature dependence of the quartz structure. Eur J
Mineral 2:63–77 Levien L, Prewitt CT, Weidner DJ (1980) Structure and elastic properties of quartz at pressure. Am Miner 65:920–930 Solovyov LA (2004) Full-profile refinement by derivative difference minimization. J Appl
Crystallogr 37:743–749 Zhernovsky IV, Kozhukhova NI, Lebedev MS (2018) Crystallochemical aspects of technological
typomorphism of quartz geomaterials during mechanoactivation. In: The collection of papers:
fundamental and applied aspects of technological mineralogy. Under the editorship of Doctor
of Geology and Mineralogy V.V. Shchiptsov. Karelian Research Center of the RAS,
Petrozavodsk, pp 97–100 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. 4
Conclusions The change of volume of silicon-oxygen tetrahedrons may be considered as an indi-
cator of the a-quartz crystal structure response to the thermal and baric influence, as
well as to the mechanoactivation. The difference of these mechanisms of influence is
demonstrated by different dependences of this parameter of a-quartz crystal structure
on the severity of exposure. Acknowledgements. The work was performed within the Federal Target Program of Research
and Development on Priority Development Fields of Science and Technology Sector in Russia
for 2014–2020, unique project number: RFMEFI58317X0063. The authors would like to express
gratitude to the Doctor of Geology and Mineralogy, Prof. N.V. Zubkova (MSU) for meaningful
consultations. 410 I. Zhernovsky and V. Strokova References If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Effect of Earthquake on the Landscape
of Jiuzhaigou-Huanglong Travertine
and Its Restoration F. Dong(&), Q. Dai, Q. Li, F. Wang, and Y. Luo Key Laboratory of Solid Waste Treatment and Resource Recycle,
School of Environment and Resource, Southwest University of Science
and Technology, Mianyang, People’s Republic of China
fqdong2004@163.com Key Laboratory of Solid Waste Treatment and Resource Recycle,
School of Environment and Resource, Southwest University of Science
and Technology, Mianyang, People’s Republic of China
fqdong2004@163.com Abstract. The
hydrogeology
and
hydrochemistry-calcium
characteristics,
microbial community changes, and dam structure geophysical exploration for
the core heritage sites of Jiuzhaigou after the “8.8” earthquake were investigated
and analyzed. The results indicated that the hydrogeology has undergone sig-
nificant changes, especially the Nuorilang Waterfall and the Sparkling Lake. The
surface hydrodynamic balance has been broken by the earthquake, which may
cause a chain reaction such as travertine deposition, travertine erosion, and
microbial community change. The Nuorilang Waterfall/Sparkling Lake dam and
the cascade waterfall are seriously affected by the earthquake. The results of the
profile study and geophysical exploration show that the surface fissures, cracks,
subsidence pipes, and other potential geological disasters are significant. The
risk of secondary collapse of the dam body and the cascade waterfall is high. It
also shows that the plant and microbes in Jiuzhaigou travertine participate in
higher deposition than Huanglong. The monitoring results show that both
Jiuzhai and Huanglong have extremely high levels of prokaryotic and eukaryotic
algae, and Jiuzhaigou is higher than Huanglong. It is suggested that on the basis
of scientific argumentation, scientific restoration of the slope and dam break of
the Sparkling Lake damage will be carried out so that the original water storage
function of the Spark Sea will be restored as soon as possible. Finally, the
Nuorilang Waterfall and Sparkling Lake Conservation Remediation Plan were
proposed and part of demonstration implementations was carried out. Keywords: Jiuzhai earthquake Travertine hydrogeology
Algae community Hydrochemical characteristics Geophysical exploration © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 413–414, 2019. https://doi.org/10.1007/978-3-030-22974-0_100 414 F. Dong et al. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 415–418, 2019.
https://doi.org/10.1007/978-3-030-22974-0_101 1
Introduction Interest in pathogenic crystallization is growing every year, which is primarily due to
the wide prevalence of diseases associated with stone formation, such as urolithiasis. Very few works are devoted to the influence of bacteria, viruses and micromycetes on
stone formation in the human body. Thus, Sagorika et al. (2013) described a patient
with renal aspergillosis along with urolithiasis, Zhao et al. (2014) reported on the
crystallization of calcium oxalate in the presence of E. coli. Other studies have noted
the initiation of calcium oxalate crystallization and aggregation in the presence of
E. coli. Most of the known works are devoted to the so-called infectious renal stones,
consisting mainly of struvite, and sometimes containing hydroxylapatite and brushite. A. Izatulina1(&), M. Zelenskaya2, and O. Frank-Kamenetskaya1 A. Izatulina1(&), M. Zelenskaya2, and O. Frank-Kamenetskaya1 1 Department of Crystallography, St. Petersburg State University,
St. Petersburg, Russia
alina.izatulina@mail.ru 1 Department of Crystallography, St. Petersburg State University,
St. Petersburg, Russia
alina.izatulina@mail.ru
2 Department of Botany, St. Petersburg State University, St. Petersburg, Russia 2 Department of Botany, St. Petersburg State University, St. Petersburg, Russia Abstract. The presence and study of the species composition of bacterial and
fungal colonies in renal stones was determined. It was shown that the presence
of microorganisms depends on the phase composition of the renal stone. No
microbial colonies were detected in oxalate stones. Under the action of the acid-
producing bacterial and fungal colonies, secondary crystallization of calcium
oxalates (whewellite and weddellite) on phosphate aggregates can occur. Keywords: Renal stones Calcium oxalates Crystallization
Phosphate renal stones Microorganisms Keywords: Renal stones Calcium oxalates Crystallization
Phosphate renal stones Microorganisms Keywords: Renal stones Calcium oxalates Crystallization
Phosphate renal stones Microorganisms Effect of Earthquake on the Landscape
of Jiuzhaigou-Huanglong Travertine
and Its Restoration Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches The study was conducted using 21 samples of renal stones of different composition: 10
– oxalate, 5 – urate, and 6 – phosphate stones. The substance of renal stones was sieved on the Czapek-Dox medium, potato
glucose agar and Saburo medium to detect the presence and to determine the species
composition of microorganisms. Preparation of nutrient media was carried out in
accordance with GOST (GOST 9.048-89). The media was sterilized in an autoclave
after preparation. After incubation period (20 days, 1 month), the samples were
microscoped and examined in accordance with identifier (de Hoog and Guarro 1995). Powder x-ray diffraction (PXRD) studies were carried out using the Rigaku
«MiniFlex II» diffractometer (CuKa radiation of wavelength k = 1.54178 Å, X-ray A. Izatulina et al. 416 tube parameters were 30 kV/15 mA; highspeed solid state energy-dispersive detector
LYNXEYE was used). X-ray diffraction patterns were collected at room temperature in
the range of 2h = 5–50° with a step of 0.02° 2h and a counting time of half second per
data point, the specimens were rotated 30 times per second during the data collection. 3
Results and Discussion As the result of urate and oxalate renal stones sieving, the detection of micromycetes on
the surface of the nutrient medium was random, and revealed a very small number of
fungal colonies; the species of micromycetes were not constant. Abundant growth of
fungal and bacterial colonies was detected on the phosphate renal stones: Cladosporium
cladosporioides, Penicillium expansum, Aspergillus niger, sporiferous light colored
fungus, Geotrichum candidum, Candida sp., Fusarium chlamydosporum, Cladospo-
rium sphaerospermum, white and pink bacterial colonies (Fig. 1). Secretion of micro-
mycetes on the surface of the nutrient medium was probably accidental, in many cases
an insignificant number of fungal colonies was detected (1–3 colonies), the species
distribution of micromycetes was not constant. There is a probability that some types of
identified micromycetes (Aspergillus niger, Cladosporium sphaerospermum, Phoma
herbarum, Penicillium purpurogemum, Penicillium expansum, Fusarium chlamy-
dosporum) accidentally hit the test renal stone material (for instance, as a result of
transportation or storage). The secretion of bacterial colonies and colonies of the fungus
Candida sp., is most likely not accidental, since a number of recent works indicate the
possibility of a bacterial biofilm formation on the surface of renal (urinary) stones
(Romanova et al. 2015). The possibility of micromycetes detection on the surface of
renal stones within the microorganisms’ biofilm requires further investigation. Fig. 1. Phosphate renal stone sample. The growth of bacterial and micromycetes colonies on the
surface of the nutrient medium in a Petri dish (∅100 mm). Fig. 1. Phosphate renal stone sample. The growth of bacterial and micromycetes colonies on the
surface of the nutrient medium in a Petri dish (∅100 mm). A colony of Aspergillus niger micromycetes was found on a nutrient medium on
one of the phosphate samples, consisting of hydroxylapatite, struvite and brushite. Aspergillus infections have grown in importance in the last years (Hedayati et al. 2007). Oxalate crystals may be present in clinical samples, due to the high acid-
producing ability of this fungus. When Aspergillus niger is growing on a liquid nutrient
medium, it was found that acidification of the culture fluid begins almost immediately
after spore germination and continues during the whole period of mycelium active
growth. After a month of incubation (Fig. 2), under the influence of Aspergillus niger 417 Microbial Colonies in Renal Stones Microbial Colonies in Renal Stones culture, the renal stone softens and many small crystals are observed during micro-
scopy. 4
Conclusions Bacterial and fungal colonies were found on the surface of phosphate renal stones; no
microbial colonies were found in oxalate and urate stones. It has been shown for the
first time that under the influence of the microscopic fungus Aspergillus niger, sec-
ondary crystallization of calcium oxalates (whewellite and weddellite) can occur on
phosphate aggregates. Thus, the possibility of the oxalate stones formation on phos-
phate nuclei with the participation of acid-producing bacterial and fungal colonies was
shown. Acknowledgements. This work was supported by the Russian Science Foundation (no. 18-77-
00026). The XRD studies have been performed at the X-ray Diffraction Centre of St. Petersburg
State University. 3
Results and Discussion According to the results of PXRD analysis, the observed crystals turned out to
be mainly calcium oxalate dihydrate (weddellite); crystals of calcium oxalate mono-
hydrate (whewellite) are also present but in subordinate quantities. Thus, under the
influence of the Aspergillus niger culture, secondary crystallization of calcium oxalate
occurs on a phosphate renal stone. The secondary crystallization of calcium oxalates
under the action of microscopic fungi was recently described for monuments of cultural
heritage (Rusakov et al. 2016). This phenomenon may be one of the reasons for the
frequent presence of a phosphate nucleus in the center of an oxalate renal stones
(Izatulina and Yelnikov 2008; Xie et al. 2014). Fig. 2. The formation of calcium oxalate crystals in the presence of Aspergillus niger and a non-
disrupting light-colored fungus during growth on a nutrient medium in a Petri dish (potato
glucose agar was used as a nutrient medium). Fig. 2. The formation of calcium oxalate crystals in the presence of Aspergillus niger and a non-
disrupting light-colored fungus during growth on a nutrient medium in a Petri dish (potato
glucose agar was used as a nutrient medium). Bonaventura M, Gallo M, Cacchio P, Ercole C, Lepidi A (1999) Microbial formation of oxalate
films on monument surfaces: bioprotection or biodeterioration? Geomicrobiology 16:55–64
De Hoog GS, Guarro J (1995) Atlas of clinical fungi, Baarn References Bonaventura M, Gallo M, Cacchio P, Ercole C, Lepidi A (1999) Microbial formation of oxalate
films on monument surfaces: bioprotection or biodeterioration? Geomicrobiology 16:55–64
De Hoog GS, Guarro J (1995) Atlas of clinical fungi, Baarn A. Izatulina et al. 418 Hedayati MT, Pasqualotto AC, Warn PA, Bowyer P, Denning DW (2007) Aspergillus flavus:
human pathogen, allergen and mycotoxin. Microbiology 153:1677–1692 Izatulina AR, Yelnikov VY (2008) Structure, chemistry and crystallization conditions of calcium
oxalates - the main components of kidney stones. In: Krivovichev SV (ed) Minerals as
Advanced Materials I. Springer-Verlag, Heidelberg, pp 231–241 Romanova YuM, Mulabaev NS, Tolordava ER, Seregi AV, Seregin IV, Alexeeva NV,
Stepanova TV, Levina GA, Barhatova OI, Gamova NA, Goncharova SA, Didenko LV,
Rakovskaya IV (2015) Microbial communities on kidney stones. Molekulyarnaya Genetika,
Mikrobiologiya i Virusologiya 33(2):20–25 Rusakov AV, Vlasov AD, Zelenskaya MS, Frank-Kamenetskaya OV (2016) The crystallization
of calcium oxalate hydrates formed by interaction between microorganisms and minerals. In:
Frank-Kamenetskaya OV, Panova EG, Vlasov DY (eds) Biogenic-Abiogenic Interactions in
Natural and Anthropogenic Systems. Springer International Publishing, Switzerland, pp 357–
377 Sagorika P, Viswajeet S, Satyanarayan S, Manish G (2013) Renal aspergillosis secondary to
renal instrumentation in immunocompetent patient. BMJ Case Rep 2013:bcr2013200306
Xie B, Halter TJ, Borah BM, Nancollas GH (2014) Aggregation of calcium phosphate and
oxalate phases in the formation of renal stones. Cryst Growth Des 15(6):3038–3045
Zhao Z Xia Y X e J Qingsheng W (2014) Role of E coli secretion and melamine in selecti e Sagorika P, Viswajeet S, Satyanarayan S, Manish G (2013) Renal aspergillosis secondar
renal instrumentation in immunocompetent patient. BMJ Case Rep 2013:bcr2013200306 renal instrumentation in immunocompetent patient. BMJ Case Rep 2013:bcr2013200306
Xie B, Halter TJ, Borah BM, Nancollas GH (2014) Aggregation of calcium phosphate and
oxalate phases in the formation of renal stones. Cryst Growth Des 15(6):3038–3045 ie B, Halter TJ, Borah BM, Nancollas GH (2014) Aggregation of calcium phosphate and
oxalate phases in the formation of renal stones. Cryst Growth Des 15(6):3038–3045 Zhao Z, Xia Y, Xue J, Qingsheng Wu (2014) Role of E. coli-secretion and melamine in selective
formation of CaC2O4H2O and CaC2O42H2O crystals. References Cryst Growth Des 14:450–458 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Fabrication of ZnO/Palygorskite
Nanocomposites for Antibacterial Application Y. Kang, A. Hui, and A. Wang(&) Key Laboratory of Clay Mineral Applied Research of Gansu Province,
Center of Eco-Material and Green Chemistry, Lanzhou Institute
of Chemical Physics, Chinese Academy of Sciences, Lanzhou, China
aqwang@licp.cas.cn Key Laboratory of Clay Mineral Applied Research of Gansu Province,
Center of Eco-Material and Green Chemistry, Lanzhou Institute
of Chemical Physics, Chinese Academy of Sciences, Lanzhou, China
aqwang@licp.cas.cn Abstract. ZnO/palygorskite nanocomposites were synthesized by chemical
deposition and calcination process for antibacterial application. The results
indicated that ZnO nanoparticles were deposited on the surface of rod-like
palygorskite. Antibacterial evaluation confirmed that ZnO/PAL nanocomposites
presented the good antibacterial behavior against Escherichia coli and Staphy-
lococcus aureus, which was mainly attributed to the synergistic effect of ZnO
and palygorskite. Keywords: ZnO Palygorskite Nanocomposites Antibacterial application © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 419–422, 2019.
https://doi.org/10.1007/978-3-030-22974-0_102 2
Methods and Approaches Fabrication of ZnO/PAL Nanocomposites: In a typical synthesis of ZnO/PAL
nanocomposites, 2 g PAL and 20 wt% Zn(NO3)26H2O were dissolved into deionized
water, and then 10 wt% NaOH solution was added into above solution for 2 h. The
mixture was ultrasonically dispersed for 30 min and aged for 24 h. The powder was
collected by centrifugation and dried at 80 °C for 6 h, and finally annealed at 400 °C
for 3 h in muffle furnace. Characterization: The products were characterized by X-ray diffractometer (XRD,
D/MAX-2200, Rigaku, Japan), transmission electron microscopy (TEM, JEM-
1200EX, FEI, USA) and energy dispersive spectroscopy (EDS) elemental composi-
tion analyzer. The specific surface area of the samples was evaluated by Brunauer-
Emmett-Teller analysis (BET, Micromeritics, Norcross, USA). Antibacterial Evaluation: E. coli and S. aureus were tested as a representative culture
both Gram-negative and Gram-positive bacteria, which kindly provided by China
Veterinary Culture Collection Center. The antibacterial activity of the sample was
evaluated by examining the minimum inhibitory concentration (MIC). 1
Introduction Widespread overuse of the antibiotics on animals has been faulted for creating
potentially threat for human beings (Li et al. 2014; Mckenna 2013). Antibiotic-resistant
bacterial strains emerge and pose increasing health risks, and thus new antibacterials
are urgently needed (Morrison et al. 2014). Inspired by the antibacterial property of
nanocomposites in the nature, functional application of natural clay minerals are of
great interest in academia and industry (Williams et al. 2011). It is well-known that zinc
oxide (ZnO) possesses excellent antibacterial properties against gram-positive bacteria
and gram-negative bacteria (Hui et al. 2016; Liu et al. 2019). However, it is difficult to
prevent from the aggregation of ZnO nanoparticles during preparation process, which
results in the adverse effects of the antibacterial properties. Palygorskite (PAL) as a
kind of clay minerals is common in many parts of the world, typically forming in
volcanic ash layers as rocks. Interestingly, PAL has one-dimensional rod like mor-
phology, high specific surface area and better ion-exchange capacity, which can be
adsorbed onto the bacterial cell by electrostatic adsorption. Therefore, it seems to be a
promising attempt to achieve a possible synergistic effect by combine PAL and ZnO
after loading ZnO nanoparticles on the surface of PAL. Therefore, ZnO/palygorskite
(ZnO/PAL) nanocomposites were synthesized by chemical deposition and calcination
process in this study, and the antibacterial properties against Escherichia coli (E. coli)
and Staphylococcus aureus (S. aureus) were also investigated. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 419–422, 2019. https://doi.org/10.1007/978-3-030-22974-0_102 420 Y. Kang et al. 3
Results and Discussion The XRD patterns of PAL and ZnO/PAL nanocomposites were shown in Fig. 1. The
diffraction peaks at 2h of 8.38°, 13.74°, 16.34° and 34.38° were the characteristic peaks of
PAL (Wang et al. 2015). The diffraction peaks of the obtained ZnO/PAL were corre-
sponded to a wurtzite ZnO structure (JCPDS standard card 36-1451) (Hui et al. 2016). Fig. 1. XRD patterns of PAL and ZnO/PAL nanocomposites Fig. 1. XRD patterns of PAL and ZnO/PAL nanocomposites The morphology of ZnO/PAL nanocomposites presented rod-like structure with a
rough surface, and the length and the diameter were around 200 * 300 nm and a
30 nm, respectively (Fig. 2a), which indicated that ZnO nanoparticles were success-
fully loaded onto the rod-like PAL surface to form a heterostructure. The specific
surface area of PAL and ZnO/PAL was found to be 173 m2g−1 and 56 m2g−1,
respectively. Compared with pure ZnO, this strategy could obviously improve the
specific surface area of ZnO, which was favorable to enhance the antibacterial activity Fabrication of ZnO/Palygorskite Nanocomposites for Antibacterial Application 421 of ZnO/PAL nanocomposites. The ring-type selected area electron diffraction pattern
indicated the generated ZnO possessed polycrystalline nature (Fig. 2b). What’s more,
EDS result showed the surface element compositions of the as-prepared nanocom-
posites were Mg, Al, Si, Fe and Zn (Fig. 2c). Fig. 2. (a) TEM image (inset is enlarged image), (b) selected area electron diffraction pattern
and (c) EDS result of ZnO/PAL nanocomposites Fig. 2. (a) TEM image (inset is enlarged image), (b) selected area electron diffraction pattern
and (c) EDS result of ZnO/PAL nanocomposites As illustrated in Fig. 3, the microbial colonies of E. coli was visible when the
concentration of sample was 0.25 mgmL−1, therefore, the MIC value of sample
against E. coli was 0.5 mgmL−1. By contrast, there was small S. aureus colonies
appeared in the plate when the sample contacts with S. aureus (Fig. 3i), and thus the
MIC value of sample against S. aureu was 2.5 mgmL−1. Fig. 3. (a, f) positive control of E. coli and S. aureus, E. coli treated by ZnO/PAL nanocomposites
with various concentrations (b) 2.5 mgmL−1, (c) 1.5 mgmL−1, (d) 0.5 mgmL−1, (e) 0.25 mgmL−1
and S. aureus (g) 5.0 mgmL−1, (h) 2.5 mgmL−1, (i) 1.0 mgmL−1, (j) 0.5 mgmL−1, respectively Fig. 3. (a, f) positive control of E. coli and S. aureus, E. 3
Results and Discussion coli treated by ZnO/PAL nanocomposites
with various concentrations (b) 2.5 mgmL−1, (c) 1.5 mgmL−1, (d) 0.5 mgmL−1, (e) 0.25 mgmL−1
and S. aureus (g) 5.0 mgmL−1, (h) 2.5 mgmL−1, (i) 1.0 mgmL−1, (j) 0.5 mgmL−1, respectively In fact, PAL as a natural carrier with large special area, could absorb the bacterial
cell by electrostatic adsorption. The reason for the synergistic antibacterial effect of
ZnO/PAL nanocomposites was mainly due to the active factor of ZnO, which gener-
ated reactive oxygen species such as OH, O2
1, O2
2−and H2O2 (Hui et al. 2016; Liu et al. 2019; Ma et al. 2015). Incorporation of ZnO not only enhanced the antibacterial
activity of natural PAL, but also reduced the cost of the preparation process, as well as
efficiently realized the functional utilization of natural clay mineral resources. 422 Y. Kang et al. 4
Conclusions In summary, ZnO/PAL nanocomposites were synthesized by chemical deposition and
calcination process. The ZnO/PAL nanocomposites exhibited the excellent antibacterial
activity against E. coli and S. aureus, and the MIC values for E. coli and S. aureus were
0.5 and 2.5 mgmL−1, respectively. Acknowledgements. This work was financially supported by the Major Projects of the National
Natural Science Foundation of Gansu, China (18JR4RA001). cknowledgements. This work was financially supported by the Major Projects of the Nationa
atural Science Foundation of Gansu, China (18JR4RA001). g
y
pp
y
j
j
Natural Science Foundation of Gansu, China (18JR4RA001). Bacterial Oxidation of Pyrite Surface S. Lipko1(&), I. Lipko2, K. Arsent’ev2, and V. Tauson1 1 Vinogradov Institute of Geochemistry SB RAS, Irkutsk, Russia
slipko@yandex.ru
2 Limnological Institute SB RAS, Irkutsk, Russia 1 Vinogradov Institute of Geochemistry SB RAS, Irkutsk, Russia
slipko@yandex.ru
2 Limnological Institute SB RAS, Irkutsk, Russia Abstract. The article considers the study of the role of bacteria in the surface
oxidation of pyrite. The experiment provided the data on characteristic mor-
phological changes of the surface and the first data on influence of a non-
autonomous phase (NP) on bacterial oxidation. Keywords: Pyrite Iron-oxidizing bacteria SEM-EDAX
Non-autonomous phase Surface References Hui AP, Liu JL, Ma JZ (2016) Synthesis and morphology-dependent antimicrobial activity of
cerium doped flower-shaped ZnO crystallites under visible light irradiation. Colloid Surf A:
Physicochem Eng Aspects 506:519–525 Li XN, Robinson SM, Gupta A, Saha K, Jiang ZW, Moyano DF, Sahar A, Riley MA,
Rotello VM (2014) Functional gold nanoparticles as potent antimicrobial agents against
multi-drug-resistant bacteria. ACS Nano 8:10682–10686 u JL, Wang YH, Ma JZ, Peng Y, Wang AQ (2019) A review on bidirectional analogie
between the photocatalysis and antibacterial properties of ZnO. J Alloy Compd 783:898–91 Liu JL, Wang YH, Ma JZ, Peng Y, Wang AQ (2019) A review on bidirectional analogies
between the photocatalysis and antibacterial properties of ZnO. J Alloy Compd 783:898–918
Ma JZ, Hui AP, Liu JL, Bao Y (2015) Controllable synthesis of highly efficient antimicrobial
agent-Fe doped sea urchin-like ZnO nanoparticles. Mater Lett 158:420–423 Ma JZ, Hui AP, Liu JL, Bao Y (2015) Controllable synthesis of highly efficient antimicr
agent-Fe doped sea urchin-like ZnO nanoparticles. Mater Lett 158:420–423 Mckenna M (2013) Antibiotic resistance: the last resort. Nature 499:394–396 Morrison KD, Underwood JC, Metge DW, Eberl DD, Williams LB (2014) Mineralogical
variables that control the antibacterial effectiveness of a natural clay deposit. Environ
Geochem Health 36:613–631 Wang WB, Zhang ZF, Tian GY, Wang AQ (2015) From nanorods of palygorskite to nanosheets
of smectite via a one-step hydrothermal process. RSC Adv 5:58107–58115 Williams LB, Metge DW, Eberl DD, Harvey RW, Turner AG, Prapaipong P, Poret-Peterson AT
(2011) What makes a natural clay antibacterial? Environ Sci Technol 45:3768–3773 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches The culture of acidophilic iron-oxidizing bacteria isolated from natural habitats (sulfide
occurrences) of the Baikal area of the Irkutsk region was used for research on the bio-
oxidation of the pyrite surface. This bacterial culture was provided by the laboratory №
7 of Irkutsk scientific-research Institute of rare and precious metals and diamonds, JSC
“Irgiredmet”. Iron-oxidizing bacteria are used in laboratory tests for bacterial oxidation
of resistant iron-sulfide ores containing gold. The synthesis of pyrite crystals was performed according to the standard technique
of hydrothermal thermogradient synthesis in titanium inserts at T = 450 °C and 500 °C
and a pressure of 1 kbar (Tauson et al. 2008). In the synthesis of pyrite Fe+S charge was
used, the composition of the surface non-autonomous phase was regulated by the
activity of sulfur depending on Fe/S ratio. The obtained crystals were up to 5 mm in
size. Pyrite, obtained at high sulfur activity, contains virtually no NP on the surface. At
lower sulfur activity, a layer of NP up to *500 nm thick with a base composition
similar to pyrrhotite, but with different forms of sulfur, is formed: Fe2+ [S, S2, Sn]2−
(Tauson et al. 2008). These surface formations are able to absorb cationic impurities
and oxysulfide anions. y
To conduct research on pyrite bio-oxidation, a mixture of acidophilic iron bacteria
was grown on a liquid 9 K medium at room temperature and constant stirring within
5 days. In eight 250 ml conical flasks with 50 ml of 9 K medium (without FeSO4)
there were placed pieces of polished pyrite (4 flasks) and 5–6 pieces of pyrite with
nonautonomous phases (4 flasks). The medium and pyrite flasks were sterilized at
0.5 atm. for 10 min to minimize pyrite oxidation. After cooling the medium, 6 flasks
were inoculated with 5-day bacterial culture. Previously, the culture was centrifuged
and washed from the medium residues with iron in 0.01 m H2SO4. The concentration
of iron bacteria cells added to the medium with pyrite was about 1*107 cells/ml. The
remaining two flasks with polished pyrite and NP on the pyrite surface were used as a
control, without bacteria. Cultivation took place at room temperature and constant
stirring on a shaker (about 110 rotation/min) for three weeks. Every week 2 flasks with
different samples of pyrite were selected for further research. The flasks with control
samples were examined after 3 weeks of cultivation. 1
Introduction For many years, extensive studies have been conducted on the processes of diagenetic
redistribution of ore-forming components in the Earth’s lithosphere and the formation
of iron-containing nodules and ores, aimed at the their prospective industrial use. Most
researchers believe that the earliest microbial ecosystems were based on sulfur trans-
formations – sulfate-reduction and disproportionation (Wacey et al. 2011). The choice of pyrite as the object of study is geochemically justified by the close
connections of iron sulfides with organic matter in various environments, including
hydrothermal conditions (Lindgren et al. 2011). Although the chemistry of the processes has been studied in principle, there
remains a number of unresolved issues. The most important are proof of paleobacterial
processes and determination of their role in the formation of mineral deposits. Here the
range of opinions is very wide: from complete denial to recognition of their leading
character at the sedimentary-hydrothermal stage of ore formation (Vinichenko 2007). The morphological effects of the interaction of mineral surfaces with bacteria have not
been sufficiently investigated, which complicates interpretation of natural observations. In particular, it is unclear what effect non-autonomous phases located within the surface
layer of the crystal have on the interaction of bacterial communities with the pyrite
surface (Tauson et al. 2008, 2009a). Non-autonomous phases (NP) are nanocrystalline
objects formed in the surface layer of the crystal through interaction with its surface of
the growth medium components or contacting autonomous (classical) phases. The
experiment within the framework of present research used specially synthesized pyrite
crystals with different degrees of NP development on the surface (Tauson and Lipko
2013), with the aim to study the process of interaction between bacteria and NP and to
establish the role of surface phases in the oxidative processes initiated by the aci-
dophilic iron bacteria. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 423–426, 2019. https://doi.org/10.1007/978-3-030-22974-0_103 424 S. Lipko et al. 2
Methods and Approaches The bacterial film from the pyrite
samples were washed with 2% aqueous solution of Polysorbate Tween 80. Pyrite
crystals washed after the experiment were dried in air and analyzed on the scanning
multi-microscope SMM 2000 in atomic force mode, scanning electron microscope FEI
Quanta Company (USA) 200 with energy dispersive device EDAX for X-ray
microanalysis. 3
Results and Discussion The experiment on pyrite bio-oxidation established that the surface of polished pyrite is
less susceptible to bacterial oxidation as compared with NP-containing pyrite. Microphotographs show the surfaces of polished pyrite (roughness less than 5 nm) and
NP-containing pyrite (roughness more than 300 nm) after two weeks of bacterial
cultivation (Fig. 1). Bacterial Oxidation of Pyrite Surface 425 Fig. 1. Surface morphology of pyrite (scanning electron microscopy). a – polished surface,
b – NP-containing surface. Fig. 1. Surface morphology of pyrite (scanning electron microscopy). a – polished surface,
b – NP-containing surface. The surface with NP exhibits bacteria and characteristic traces of interaction
between bacteria and pyrite in the form of holes of different size comparable to the
sizes of bacteria. For polished pyrite, these traces are almost absent and are observed
only on the borders of scratches left from polishing. A similar result was obtained for
the pyrite surface with minimal NP development synthesized at high sulfur activity. Therefore, the activity of bacteria is associated with the structure of pyrite surface. Similar formations, but significantly smaller (nano-holes) were discovered earlier, in
the study of pyrites from the Sukhoi Log gold deposit (Irkutsk region) (Tauson et al. 2009b). This confirms the affinity of processes occurring in nature and in the
experiment. 4
Conclusions The research resulted in acquisition of data on pyrite bio-oxidation taking into account
the structure of crystal surface under the given conditions. For this purpose, crystals
with different degrees of non-autonomous phases development on the surface con-
trolled by growth conditions were synthesized and used in the experiment for the first
time. It was found that the surface of polished pyrite is less susceptible to bacterial
oxidation, as compared with pyrite containing a non-autonomous phase. The resulting
characteristic morphological changes in the surface will further be instrumental in
addressing the issues of ore genesis, as well as identifying minerals that were formed at
the initial or final stage of growth involving bacteria. Acknowledgements. We thank Alexandra Mikhailova for suppling of bacterial culture. The
research was performed within a state assignment, Project IX.125.3, No. 0350-2016-0025 and
was funded by the Federal Agency for Scientific Organizations (FASO) within the framework of
State Tasks No. 0345-2016-0003 (AAAAA16- 116122110061-6). S. Lipko et al. 426 References Lindgren P, Parnell J, Holm NG, Broman C (2011) A demonstration of the affinity between
pyrite and organic matter in a hydrothermal setting. J Geochem Trans 12(3):3–7 Tauson VL, Lipko SV (2013) Pyrite as a concentrator of gold in laboratory and natural systems: a
surface-related effect. In: Whitley N, Vinsen PT (eds) Pyrite: Synthesis, Characterization and
Uses Chapter 1. Nova Science Publisher Inc., New York, pp 1–40 Tauson VL, Babkin DN, Lustenberg EE, Lipko SV, Parkhomenko IY (2008) Surface
typochemistry of hydrothermal pyrite: electron spectroscopic and scanning probe microscopic
data I. Synthetic pyrite. J Geochem Int 46(6):615–628 Tauson VL, Kravtsova RG, Grebenshchikova VI, Lustenberg EE, Lipko SV (2009a) Surface
typochemistry of hydrothermal pyrite: electron spectroscopic and scanning probe microscopic
data II. Natural pyrite. J Geochem Int 47(3):245–258 py
Tauson VL, Lipko SV, Shchegolkov YuV (2009b) Surface nanoscale relief of mineral crystals
and its relation to non-autonomous phase formation. J Crystallogr Rep 54(7):1219–1227 Vinichenko PV (2007) Biogeology and Ore Formation. Izd-e Sosnovgeologiya, Irkutsk Wacey D, Saunders M, Brasier MD, Kilburn MR (2011) Earliest microbially mediated pyrite
oxidation in 3.4 billion-year-old sediments. J Earth Planet Sci Lett 301:393–402 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Biomimetic Superhydrophobic Cobalt
Blue/Clay Mineral Hybrid Pigments
with Self-cleaning Property
and Different Colors B. Mu1, A. Zhang1,2, and A. Wang1(&) 1 Key Laboratory of Clay Mineral Applied Research of Gansu Province,
Center of Eco-material and Green Chemistry,
Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences,
Lanzhou 730000, People’s Republic of China
aqwang@licp.cas.cn
2 Center of Materials Science and Optoelectronics Engineering,
University of Chinese Academy of Sciences, Beijing 100049,
People’s Republic of China 1 Key Laboratory of Clay Mineral Applied Research of Gansu Province,
Center of Eco-material and Green Chemistry,
Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences,
Lanzhou 730000, People’s Republic of China
aqwang@licp.cas.cn
2 Center of Materials Science and Optoelectronics Engineering,
University of Chinese Academy of Sciences, Beijing 100049,
People’s Republic of China 1 Key Laboratory of Clay Mineral Applied Research of Gansu Province,
Center of Eco-material and Green Chemistry,
Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences,
Lanzhou 730000, People’s Republic of China
aqwang@licp.cas.cn
2 Center of Materials Science and Optoelectronics Engineering,
University of Chinese Academy of Sciences, Beijing 100049,
People’s Republic of China Abstract. Inspired by self-cleaning and water-repellent properties of the lotus
leaf, biomimetic superhydrophobic cobalt blue/clay mineral hybrid pigments
were facilely fabricated based on the rough surface of hybrid pigments and the
modification with various organosilanes. The obtained hybrid pigments were
characterized using various analytical techniques. Due to the difference in the
compositions and morphologies of clay minerals, the obtained cobalt blue/clay
mineral hybrid pigments exhibited different color properties. Superhydropho-
bicity of hybrid pigments was mainly regulated by the types of organosilanes
instead of the morphologies of hybrid pigments. The sprayed coating of the
superhydrophobic hybrid pigments exhibited the excellent self-cleaning per-
formance with high water contact angle and low sliding angle. The coatings also
presented excellent environmental and chemical durability even under harsh
conditions. Therefore, the obtained biomimetic superhydrophobic cobalt
blue/clay mineral hybrid pigments may be applied in various fields, such as
anticorrosion, self-cleaning coating, etc. Keywords: Biomimetic Superhydrophobic Cobalt blue Clay minerals
Self-cleaning Keywords: Biomimetic Superhydrophobic Cobalt blue Clay minerals
Self cleaning Keywords: Biomimetic Superhydrophobic Cobalt blue Clay minerals
Self-cleaning 1
Introduction Cobalt blue (cobalt aluminate, CoAl2O4) pigment is a typical eco-friendly blue inor-
ganic pigment. Due to high refractive index, excellent chemical and thermal stability, it
can be widely applied in ceramics, paints, engineering plastics, etc. However, the high
cost of CoAl2O4 pigment severely restrains their relevant applications due to the high
price of cobalt compounds and the disadvantages of the traditional solid phase method © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 427–431, 2019. https://doi.org/10.1007/978-3-030-22974-0_104 428 B. Mu et al. (Armijo 1969). In addition, most of the common application fields of CoAl2O4 pigment
are hydrophilic, which may be easily contaminated by dark liquid, oil stains, blot in our
daily life. Therefore, it is very necessary to develop low-cost CoAl2O4 pigment with
and self-cleaning ability. Inspired by the unique water-repellent surfaces of the lotus leaf in the natural world
(Barthlott and Neinhuis 1997), the design of superhydrophobic surfaces has become the
focus in both fundamental research and industrial applications by construction of rough
surface structure and modification using materials with low surface free energy. Recently, substrate-based inorganic hybrid pigments composed of an inorganic sub-
strate coated with inorganic pigment nanoparticles have attracted increasingly attention. Our groups successfully prepared CoAl2O4 hybrid pigment after incorporation of
different clay minerals (Mu et al. 2015; Zhang et al. 2017). Incorporation of clay
minerals greatly decreased the production cost and calcining temperature for formation
of spinel CoAl2O4, as well as preventing from the aggregation of CoAl2O4 nanopar-
ticles after being uniformly anchored on the surface of clay minerals. Based on the
rough surface of the hybrid pigments, it could realize the superhydrophobic modifi-
cation of hybrid pigments using materials with low surface free energy. In this study,
different CoAl2O4 hybrid pigments derived from kaoline (Kaol), palygorskite (Pal),
halloysite (Hal) and montmorillonite (Mt) were prepared and modified using different
organosilanes including octyl triethoxysilane (OTES), dodecyl trimethoxysilane
(DTMS),
hexadecyltriethoxysilane
(HTES)
and
perfluoroctyl
trimethoxysilane
(PFOTMS), and the effect of the types organosilanes and the morphologies of hybrid
pigments on the superhydrophobic properties was comparatively studied. 3
Results and Discussion Table 1. Color parameters, water contact angle (CA) and sliding angle (SA) of superhydropho-
bic hybrid pigments before and after being treated at various conditions Table 1. Color parameters, water contact angle (CA) and sliding angle (SA) of superhydroph
ic hybrid pigments before and after being treated at various conditions Table 1. Color parameters, water contact angle (CA) and sliding angle (SA) of superhydropho-
bic hybrid pigments before and after being treated at various conditions Samples
Conditions
L*
a*
b*
CA/°
SA/°
Kal-HP-HTES
-
38.1 2.6
−63.8 164.2 1.0
98% H2SO4
37.4 2.6
−63.6 163.8 1.2
3 M NaOH
38.1 2.6
−63.8 163.3 1.3
UV for 3 days 38.0 2.6
−63.9 164.0 1.1
Pal-HP-HTES
-
16.2 −21.1 −23.5 148.1 2.4
98% H2SO4
15.8 −20.8 −26.2 148.2 2.3
3 M NaOH
16.7 −21.3 −24.8 147.5 2.4
UV for 3 days 16.9 −22.2 −23.3 148.3 2.4
Hal-HP-HTES
-
55.8 −2.3
−55.0 151.2 1.2
98% H2SO4
55.6 −2.4
−54.6 151.6 1.5
3 M NaOH
55.8 −2.8
−53.7 151.8 1.8
UV for 3 days 55.2 −2.6
−54.2 150.8 1.2
Mt-HP-HTES
-
24.5 −21.2 −32.3 156.4 3.7
98% H2SO4
23.2 −21.3 −33.2 155.6 3.4
3 M NaOH
24.6 −21.4 −32.7 157.5 3.1
UV for 3 days 21.2 −20.4 −30.5 154.8 3.3
Kal-HP-OTES
-
38.1 2.5
−63.8 135.1 4.0
Kal-HP-DTES
-
38.1 2.6
−63.8 151.2 3.0
Kal-HP-PFOTMS -
38.0 2.5
−63.7 165.2 2.0 The color parameters, CA and SA of superhydrophobic hybrid pigments before and
after being treated at various conditions were summarized in Table 1. It was found that
superhydrophobic hybrid pigments derived from different clay minerals presented
different color properties (Fig. 1), which might be attributed to the difference in the
compositions of clay minerals, especially Fe element. Among the employed clay
minerals, the higher content of Fe element was observed in Pal and Mt than Kal and
Hal, which decreased the color parameters of hybrid pigments. By contrast, hybrid
pigment prepared using Hal exhibited the optimum color properties, and hybrid pig-
ments obtained from Kal came second. In addition, the water contact angle and sliding
angle of different hybrid pigments derived from different clay minerals had no obvious
difference after being modified using HTES (Fig. 1). Except for Pal-HP-HTES, the
values of CA and SA of them were higher than 150º and below 5º, respectively. 2
Methods and Approaches Cobalt blue/hybrid pigments were prepared according to the similar procedure reported
in our previous study (Zhang et al. 2017). Typically, 2.91 g of Co(NO3)2∙6H2O, 7.50 g
of Al(NO3)3∙9H2O and 1.09 g of clay minerals were added to 50 mL of water under
magnetic stirring at 150 rpm for 30 min, and then 3 M NaOH was added dropwise into
above mixture until the pH was reached to 10. The suspension was continuously stirred
for 2 h at room temperature, and then the solid products were collected by centrifu-
gation, washed with water and directly calcined at 1100 °C for 2 h with a rate of
10 °C/min. Next, the superhydrophobic modification of hybrid pigments was conducted
in the ammonia saturated ethanol solution (Zhang et al. 2018). 0.54 g of organosilanes
and 1.5 g of hybrid pigments with different weight ratios were firstly added into 45 mL
above ethanol solution and stirred for 1 h at room temperature, and then 4.00 g of water
was injected quickly into the solution and stirred for 24 h at room temperature. The solid
products were finally washed with ethanol for three times and dried in an oven at 60 °C. The obtained samples were labeled as clay mineral-HP-organosilanes according to the
involved clay minerals and organosilanes. Biomimetic Superhydrophobic Cobalt Blue/Clay Mineral Hybrid Pigments 429 3
Results and Discussion Although the hybrid pigments were treated at various conditions, the color properties,
CA and SA almost kept stable, indicating the excellent environmental and chemical
durability. Furthermore, superhydrophobic Kal-HP modified with various organosi-
lanes presented different CA and SA. With the increase in the carbon chain length, the
values of SA increased while the SA values decreased. Meanwhile, the incorporation of B. Mu et al. 430 fluorine atom in organosilanes also favored enhancing the superhydrophobic properties
of hybrid pigments. Fig. 1. Digital photos of (a) Hal-HP-HTES, (b) Mt-HP-HTES, (c) APT-HP-HTES and (d) Kal-
HP-HTES Fig. 1. Digital photos of (a) Hal-HP-HTES, (b) Mt-HP-HTES, (c) APT-HP-HTES and (d) Kal-
HP-HTES 4
Conclusions Superhydrophobic cobalt blue/clay mineral hybrid pigments with self-cleaning prop-
erty and different colors were successfully prepared by modifying using organosilanes
based on the rough surface. The color and superhydrophobicity were closely related to
the compositions of clay minerals and the types of organosilanes. Acknowledgements. The authors are grateful for financial support of the Major Projects of the
National Natural Science Foundation of Gansu, China (18JR4RA001), and the Youth Innovation
Promotion Association of CAS (2017458). arthlott W, Neinhuis C (1997) Purity of the sacred lotus, or escape from contamination in
biological surfaces. Planta 202:1–8 © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 432–435, 2019.
https://doi.org/10.1007/978-3-030-22974-0_105 1
Introduction Over the past decades, numerous studies have shown that Si is an essential element and it
affects human health. Silicon is present in the body as a trace element, but so far its
biochemical function has not been confirmed by experimental data. Bioavailable silicon
usually enters the body from solutions. Most easily it diffuses through the membranes
and penetrates into the circulatory system in the form of orthosilicate acid, which is
present in water, beer and some beverages. Among food products, the most significant
sources of silicon are the products of plant origin: wheat, rice, oats, and barley. Despite
the fact that vegetable food has a high content of silicon, its bioavailability is very limited
due to the poor solubility of the forms of silicon present in the plants (Farooq 2015). In this paper we present amorphous silica found in the composition of pathological
formations - calcified heart valves. To explain the possible route of ingestion of
amorphous silica, we considered one of the most common food crops for silicon
consumption - rice. Keywords: Silicon dioxide Scanning and transmission electron microscopy
Mineralized heart valves Bioavailability Keywords: Silicon dioxide Scanning and transmission electron microscopy
Mineralized heart valves Bioavailability References Armijo JS (1969) The kinetics and mechanism of solid-state spinel formation—a review and
critique. Oxid Met 1:171–198 Barthlott W, Neinhuis C (1997) Purity of the sacred lotus, or escape from contaminatio
biological surfaces. Planta 202:1–8 Mu B, Wang Q, Wang AQ (2015) Effect of different clay minerals and calcination temperature
on the morphology and color of clay/CoAl2O4 hybrid pigments. RSC Adv 5:102674–102681 p
gy
y
y
p g
Zhang AJ, Mu B, Luo ZH, Wang AQ (2017) Bright blue halloysite/CoAl2O4 hybrid pigments:
preparation, characterization and application in water-based painting. Dyes Pigm 139:473–481 Zhang AJ, Mu B, Luo ZH, Wang AQ (2017) Bright blue halloysite/CoAl2O4 hybrid pigments:
preparation, characterization and application in water-based painting. Dyes Pigm 139:473–481 Zhang AJ, Mu B, Hui AP, Wang AQ (2018) A facile approach to fabricate bright blue heat-
resisting paint with self-cleaning ability based on CoAl2O4/kaoline hybrid pigment. Appl
Clay Sci 160:153–161 Zhang AJ, Mu B, Hui AP, Wang AQ (2018) A facile approach to fabricate bright blue heat-
resisting paint with self-cleaning ability based on CoAl2O4/kaoline hybrid pigment. Appl
Clay Sci 160:153–161 431 Biomimetic Superhydrophobic Cobalt Blue/Clay Mineral Hybrid Pigments Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Silicon Dioxide in Mineralized Heart Valves A. Titov1,2(&), V. Zaikovskii1, and P. M. Larionov1,3 1 National Research University, Novosibirsk, Russia
titov@igm.nsc.ru
2 Sobolev V.S. Institute of Geology and Mineralogy of SB RAS,
Novosibirsk, Russia
3 Boreskov Institute of Catalysis of SB RAS, Novosibirsk, Russia 1 National Research University, Novosibirsk, Russia
titov@igm.nsc.ru
2 Sobolev V.S. Institute of Geology and Mineralogy of SB RAS,
Novosibirsk, Russia
3 Boreskov Institute of Catalysis of SB RAS, Novosibirsk, Russia Abstract. This study showed that silicon dioxide of plant origin penetrated into
the human body unchanged and was transferred through the blood to the heart,
where absorbed by the pathological hydroxyapatite of mineralized heart valves. 2
Methods and Approaches We studied intraoperative material, which included mineralized aortic and mitral valves
of the heart, obtained from patients with acquired heart defects of rheumatic and septic
genesis (Titov et al. 2016). Siliceous formations found in the calcifications of the heart
valves were compared with the siliceous formations of plant origin: rice straw and husk. The inventory of methods for structural and elemental analyses included high-
resolution transmission electron microscopy (HR TEM), scanning electron microscopy
(SEM), electron diffraction, and energy-dispersive X-ray spectroscopy (EDX). Elec-
tron microscopy was performed using a JEM2010 transmission electron microscope 433 Silicon Dioxide in Mineralized Heart Valves (acceleration voltage 200 kV, resolution 1.4 Å) equipped with an EDAX EDS detector
(spectral resolution 130 eV) and a TESCAN MIRA3 scanning electron microscope
with an Oxford EDS detector (resolution 128 eV) and built-in INKA ENERGY
software. (acceleration voltage 200 kV, resolution 1.4 Å) equipped with an EDAX EDS detector
(spectral resolution 130 eV) and a TESCAN MIRA3 scanning electron microscope
with an Oxford EDS detector (resolution 128 eV) and built-in INKA ENERGY
software. 3
Results and Discussion Our investigation relied on the data obtained in the studies of the calcified formations
on heart valves and their bioprostheses (Titov et al. 2016). The presence of silicon at
the trace level was observed in the EDS analyzes of calcified heart valves. The studies
of the dispersed material of heart valve calcifications by means of transmission electron
microscopy revealed amorphous particles of rounded shape about 100 nm in diameter,
along with hydroxyapatite nanocrystals (Fig. 1). The EDS spectra of these amorphous
particles revealed Si, O, and C (carbon deposition), which corresponded to silicon
dioxide (Fig. 1). Fig. 1. Electron microscopic image (TEM) of amorphous silica particles among the dispersed
calcified mineralized substance of a heart valve. Insert: EDX spectrum from one of the particles
of silicon dioxide Fig. 1. Electron microscopic image (TEM) of amorphous silica particles among the dispersed
calcified mineralized substance of a heart valve. Insert: EDX spectrum from one of the particles
of silicon dioxide As can be seen on the SEM images and EDS spectra of rice straw and husk
(Fig. 2.), their surfaces were covered with a thin layer of silicon dioxide. After
annealing the rice straw and husk at a temperature of 750 °C, white silica powder
remained. The examination of the powder preparation by means of transmission
electron microscopy showed that the rice substrate was represented by the rounded
particles of amorphous silica with a diameter of about 50 nm (Fig. 3). The similarity of
the amorphous particles of the rice substrate with the amorphous particles from heart
valve calcifications in chemical composition and structure seemed quite obvious to us. 434 A. Titov et al. Fig. 2. SEM morphology of rice straw and husk. On the insert is the EDX spectrum from area 2
in the image. Fig. 2. SEM morphology of rice straw and husk. On the insert is the EDX spectrum from area 2
in the image. Plants accumulate Si in the form of biogenic (phytolytic) silicon dioxide from soil
solutions. It becomes included in plant tissues as a structural component imparting
strength and rigidity to the stems. The main route of silicon intake into the organism starts
from the gastrointestinal tract. Phytolytic silica is considered to be an insoluble form of Si. However, most of Si is absorbed from solid products, therefore it is assumed that
the phytolytic silicon dioxide is destroyed and absorbed (Jugdaohsingh 2007). 3
Results and Discussion Silicon
dioxide detected by us in calcifications with nanocrystalline hydroxyapatite, remained
unchanged. Apparently silica particles were transferred to the heart valves from the
patients’ blood and precipitated on hydroxyapatite. Hydroxyapatite is known to have a
high sorption capacity (Titov et al. 2013). Perhaps a significant amount of silica can be
deposited on the bone tissue as hydroxyapatite is one of its components. Our analysis
of the chemical composition of calcified mineralized heart valves using an EDS
spectrometer detected silicon at a trace level. The Si content in calcifications is quite
significant for this trace element considering the sensitivity level of this method as 0.n
weight percent. Fig. 3. A and B.Electron microscopic images (TEM) of cytolytic silica obtained from rice straw
and husk after annealing at a temperature of 750 °C. B – the image of a fragment of A. Fig. 3. A and B.Electron microscopic images (TEM) of cytolytic silica obtained from rice straw
and husk after annealing at a temperature of 750 °C. B – the image of a fragment of A. 435 Silicon Dioxide in Mineralized Heart Valves 4
Conclusions To conclude, we suggest that dispersed phytolytic silica may penetrate through the
gastric tract into human blood in the unchanged form. Acknowledgements. This research was carried out within the State Assignment to IGM SB
RAS (project 0330-2016-0013). RAS (project 0330-2016-0013). References Farooq MA, Dietz K-J (2015) Silicon as versatile player in plant and human biology: overlooked
and poorly understood. Front Plant Sci 6:994–1023 Farooq MA, Dietz K-J (2015) Silicon as versatile player in plant and human biology: overlooked
and poorly understood. Front Plant Sci 6:994–1023 Farooq MA, Dietz K-J (2015) Silicon as versatile player in plant and human biology: overlooked
and poorly understood. Front Plant Sci 6:994–1023 p
y
Titov AT, Zaikovskii VI, Larionov PM (2016) Bone-like hydroxyapatite formation in human
blood. Int J Environ Sci Educ 11(10):3971–3984 Titov AT, Zaikovskii VI, Larionov PM (2016) Bone-like hydroxyapatite formation in human
blood. Int J Environ Sci Educ 11(10):3971–3984 (
)
Jugdaohsingh R (2007) Silicon and bone health. J Nutr Health Aging 11(2):99–110 Jugdaohsingh R (2007) Silicon and bone health. J Nutr Health Aging 11(2):99–110 itov AT, Larionov PM, Zaikovskii I (2013) Calcium phosphate mineralization of bacteria. I Proceedings of the 11th International Congress For Applied Mineralogy (ICAM), pp 9–17 Proceedings of the 11th International Congress For Applied Mineralogy (ICAM), pp 9–17 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Preparation of Macroporous Adsorbent Based
on Montmorillonite Stabilized Pickering
Medium Internal Phase Emulsions F. Wang1,2, Y. Zhu1, W. Wang1, and A. Wang1(&) 1 Key Laboratory of Clay Mineral Applied Research of Gansu Province,
Center of Eco-Material and Green Chemistry,
Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences,
Lanzhou, People’s Republic of China
aqwang@licp.cas.cn
2 College of Petroleum and Chemical Engineering, Qinzhou University,
Qinzhou, People’s Republic of China 1 Key Laboratory of Clay Mineral Applied Research of Gansu Province,
Center of Eco-Material and Green Chemistry,
Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences,
Lanzhou, People’s Republic of China
aqwang@licp.cas.cn
2 College of Petroleum and Chemical Engineering, Qinzhou University,
Qinzhou, People’s Republic of China 1 Key Laboratory of Clay Mineral Applied Research of Gansu Province,
Center of Eco-Material and Green Chemistry,
Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences,
Lanzhou, People’s Republic of China
aqwang@licp.cas.cn
2 College of Petroleum and Chemical Engineering, Qinzhou University,
Qinzhou, People’s Republic of China Abstract. A macroporous material was prepared using oil-in-water Pickering
medium internal phase emulsions (Pickering MIPEs) as template. The obtained
macroporous materials with interconnected pore structure exhibited good
adsorption capacities to Ce (III) and Gd (III) in water. The adsorption process
could be achieved in 30 min, and the maximum adsorption capacities reached
230.64 mg/g for Ce (III) and 240.49 mg/g for Gd (III). Furthermore, the
macroporous
monolith
exhibited
excellent
reuseability
after
consecutive
adsorption-desorption cycles. Keywords: Pickering emulsion Medium internal phase emulsions
Rare metal Adsorption Porous material 2
Methods and Approaches Typically, macroporous carboxymethyl cellulose-g-poly(acrylamide)/montmorillonite
(CMC-g-PAM/MMT) monolith was prepared based on Pickering MIPEs, which sta-
bilized with 5% of Mt and 4% of Tween-20 (Wang et al. 2017). The obtained
monolithic polymers were washed with acetone for 12 h and then immersed into 0.5 M
NaOH aqueous alcohol solution (Vwater/Valcohol = 3/7) for 24 h to transfer the amide
group to carboxyl. The effect of the adsorption time and the initial concentration on the adsorption
capacities were conducted according to the following procedure: 20 mg porous
adsorbents were added into 25 mL Ce (III) and Gd (III) solution and shocked in a
thermostatic shaker at 120 rpm and 30 °C for a given time. After the adsorption, the
adsorbents were separated and the concentrations of the Ce (III) and Gd (III) were
determined via UV-vis spectrophotometer using the chlorophosphonazo and azo arsine
as the complexing agents, respectively. The adsorption capacities qe (mg/g) of the
porous monolithic adsorbents were calculated according to the following equation: qe ¼ C0 Ce
ð
ÞV
m
ð1Þ ð1Þ m where C0 and Ce were the initial and equilibrium concentrations of Ce (III) and Gd
(III) (mg/L), V (L) was the volume of the Ce(III) and Gd(III) solution. The reusability studies were performed as follows: The adsorbents were desorbed
by immersing 30 mL hydrochloric acid solution (0.5 M) for 2 h after adsorption, and
then regenerated with 0.5 M NaOH solution. Finally, the adsorbents were filtered and
washed to reach neutral using distilled water before next adsorption process. The
adsorption-desorption cycle was repeated five times. Preparation of Macroporous Adsorbent Preparation of Macroporous Adsorbent 1
Introduction Compared with the conventional emulsion, Pickering emulsions exhibit peculiar long-
term stability against droplet coalescence, in which the solid particles adsorbed at the
oil-water interface act as a mechanical barrier to protect the dispersion phase liquid
drops from the coalescence with the continuous phase (Tajik et al. 2017). Due to the
unique properties, Pickering emulsions have been used as template to prepare porous
polymer monoliths (Briggs et al. 2015). However, the dispersion phase of oil/water
Pickering high internal phase emulsions mostly need a large amount of organic solvent,
and thus it is indispensible to develop Pickering MIPEs by replacing poisonous organic
solvent with low-cost and eco-friendly plant oil and reducing the internal phase volume. y p
g
p
Herein, monolithic macroporous materials were fabricated for adsorption of
Ce (III) and Gd (III) based on Pickering MIPEs template, which was composed of the
stabilizer of montmorillonite (Mt) and Tween-20 and the continuous phase of flaxseed
oil. The effects of adsorption parameters including initial concentration and contact
time on the adsorption properties were investigated, and the reusable performance of
the adsorbent was also evaluated. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 436–439, 2019. https://doi.org/10.1007/978-3-030-22974-0_106 437 3
Results and Discussion The representative images of macroporous monolith prepared by Pickering-MIPEs with
5% Mt and 4% Tween-20 as the stabilizer were shown in Fig. 1. The emulsions didn’t
flow in the inverted plastic centrifuge tube (Fig. 1a), indicating that the oil droplets
were closely packed and the formed emulsion was a typical gel emulsion. The
macroporous polymer monoliths of CMC-g-PAM/MMT were synthesized by free
radical polymerization using APS as the initiator. The prepared wet monoliths were cut
into pieces and Soxhlet extracted using acetone to remove the oil phase and surfactant,
and then immerged into NaOH alcohol solution to complete hydrolysis of amide
groups. Finally, the white monoliths of CMC-g-PAM/MMT were obtained after the
dehydrated with acetone and dried in oven at 40 °C (Fig. 1b). The surface morphology
of CMC-g-PAM/MMT was shown in Fig. 1c, and it presented a hierarchical pore
structure with high connectivity. According to the statistical result of Image-Pro Plus
6.0 software, the average pore size of the macropore and the pore throat were 1.43 lm
and 0.39 lm, respectively. Furthermore, the as-prepared porous materials exhibited
narrow macropores and pore throats size distribution (Fig. 1d). 438 F. Wang et al. Fig. 1. Digital photographs of (a) the as-prepared Pickering MIPEs, (b) CMC-g-PAM/MMT
monolith, (c) SEM image of porous CMC-g-PAM/MMT monolith and (d) pore size distribution
of porous CMC-g-PAM/MMT Fig. 1. Digital photographs of (a) the as-prepared Pickering MIPEs, (b) CMC-g-PAM/MMT
monolith, (c) SEM image of porous CMC-g-PAM/MMT monolith and (d) pore size distribution
of porous CMC-g-PAM/MMT CMC-g-PAM/MMT monoliths were employed to remove of Ce (III) and Gd
(III) from water. As shown in Fig. 2a and b, the adsorption capacities increased with the
increase in the initial metal ions concentrations until the adsorption saturation was
reached. The maximum adsorption capacities of the macroporous monoliths were
230.64 mg/g for Ce (III) and 240.39 mg/g for Gd (III). The higher adsorption capacity
might be due to the sufficient functional groups and the highly interconnected pore
structure. The effect of contact time of on the adsorption behavior was depicted in
Fig. 2c and d. It was obvious that the porous monolithic adsorbent showed fast
adsorption rate for Ce (III) and Gd (III), and the adsorption equilibrium could be reached
within 30 min and 25 min Ce (III) and Gd (III), respectively. 4
Conclusions Macroporous polymer monoliths of CMC-g-PAM/MMT were successfully synthesized
by free radical polymerization based on based on Pickering MIPEs stabilized with 5%
of Mt and 4% of Tween-20. The as-prepared macroporous polymer monoliths pos-
sessed a hierarchical pore structure and highly interconnection, which favored
enhancing the adsorption properties to Ce (III) and Gd (III), such as high adsorption
capacity, quick adsorption rate, and good reusability. Acknowledgements. The authors are grateful for financial support of the Major Projects of the
National Natural Science Foundation of Gansu, China (18JR4RA001) and the National Natural
Science Foundation of China (21706267). Science Foundation of China (21706267). 3
Results and Discussion The macro-pores allowed
fast and efficient mass transport, as well as provided sufficient contact between active
groups and adsorbents, while the pore throats contributed to a high specific surface area. Fig. 2. Effect of the initial concentration of (a) Ce (III) and (b) Gd (III) on the adsorption
capacity of porous monolith. Adsorption kinetic curves of the porous monolith for (c) Ce
(III) and (d) Gd (III). Fig. 2. Effect of the initial concentration of (a) Ce (III) and (b) Gd (III) on the adsorption
capacity of porous monolith. Adsorption kinetic curves of the porous monolith for (c) Ce
(III) and (d) Gd (III). 439 Preparation of Macroporous Adsorbent References Tajik S, Nasernejad B (2017) Surface modification of silica-graphene nanohybrid as a novel
stabilizer for oil-water emulsion. Korean J Chem Eng 34:2488–2497 Briggs NM, Weston JS, Li B, Venkataramani D, Aichele CP, Harwell JH, Crossley SP (2015)
Multiwalled carbon nanotubes at the interface of Pickering emulsions. Langmuir 31:13077–
13084 Wang F, Zhu YF, Wang WB, Zong L, Lu TT, Wang AQ (2017) Fabrication of CMC-g-PAM/Pal
superporous polymer monoliths via eco-friendly Pickering-MIPEs for superior adsorption of
methyl violet and methylene blue. Front Chem 5:33 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 443–444, 2019.
https://doi.org/10.1007/978-3-030-22974-0_107 1
Introduction In addition to the problems of each civilization, humanity is faced with the urgent need
to solve planetary problems. At the end of XX century, the first terrible signs of the
deterioration of the quality of the biosphere had already appeared as a result of the
development of man-made civilization and the installation to conquer nature started. Smog over large cities, deforestation and the onset of deserts, depletion of the soil
and basins of many rivers, a decrease in the number of fish and wild animals – all this
worried people at the beginning of the twentieth century. Depletion of the Land Resources and Its Effect
on the Environment M. Abou Zahr Diaz, M. A. Alawiyeh, and M. Ghaboura(&) Department of Mineral Developing and Oil & Gas Engineering,
Engineering Academy, RUDN University, Moscow, Russia
mostafa_ghab@live.com Department of Mineral Developing and Oil & Gas Engineering,
Engineering Academy, RUDN University, Moscow, Russia
mostafa_ghab@live.com Abstract. Resources depletion refers to the situation where the consumption of
natural resources is faster than it can be replenished. In order to achieve economic
growth, developing countries are abusing their lands on the grounds of economic
interests. Population Explosion is acting as a catalyst for resources depletion. It
seems evident that developing countries pursuing rapid economic growth disre-
gard environmental concerns. The natural resources contribute at large to the
economic development of a nation. Consumption pattern if not addressed will lead
to irreversible climate change and declined economic growth, as a result of
increased social, economic, and environmental costs and decreased productivity. Resource utilization has always been part of human history; however, the accel-
eration of economic growth activities together with the pursuit of an urgent eco-
nomic development is the core cause of resources overexploitation. Consumption
pattern will lead to irreversible climate change and declined economic growth. Keywords: Resources Economics Population Utilization
Depletion of the soil Keywords: Resources Economics Population Utilization
Depletion of the soil Keywords: Resources Economics Population Utilization
Depletion of the soil 2
Methods and Approaches A no less formidable problem is the ecological catastrophe approaching the planet. At
present, mankind produces organic waste in an amount of two thousand times more
than the waste volumes of the rest of the biosphere. Obviously, the violation of this
equilibrium caused a whole complex of complex problems. Man, unlike all living
things, is not strictly bound by the environmentally friendly conditions of his being, in 444 M. Abou Zahr Diaz et al. a certain sense he was always going against nature, not adapting to it, but changing it in
accordance with its needs (Satterthwaite 2009). 4
Conclusions The current generation cannot help thinking about future children and grandchildren,
who are to continue to carry the baton of history. Unfortunately, our civilization largely
lives at the expense of the future, exhausting irreplaceable resources (oil, gas), polluting
water, air and soil with its imperfect technologies, preserving many archaic social
structures, sowing seeds of national and religious hatred that will sprout another century. Based on the above, it can be concluded that only the efforts of the entire world
community can prevent an environmental catastrophe that threatens all life on Earth. 3
Results and Discussion The demographic problem has become global long ago. In 1987, the five billionth
inhabitant of the planet was born, and the growth rate is now such that every second the
number of people on Earth is increased by three people (Anderson 2012). According to
the figurative expression of scientists, the Earth is now “biting man” and it is quite
natural to expect a demographic collapse in the near future, that is, a sufficiently sharp
decline in population. It can be caused by global hunger, depletion of mineral resources and soil, poor
drinking water, thermal overheating of the surface, etc. 1
Introduction Kabwe town is the worst polluted place in Africa due to mining and smelting of Pb and
Zn ores. Orthophosphate has been receiving a lot of attention as stabilization agent for
heavy metals, In order to reduce dispersion and mobility of Pb metal from the slag, we
suggests treatment by adding phosphoric fertilizer (Triple super phosphate: TSP) which
is effective and locally available. Geochemical Behavior of Heavy Metals During
Treatment by Phosphoric Fertilizer
at a Dumping Site in Kabwe, Zambia H. Kamegamori1(&), K. Lawrence1, T. Sato2, and T. Otake2 1 Graduate School of Engineering, Hokkaido University, Sapporo, Japan
skj-9mm@eis.hokudai.ac.jp
2 Faculty of Engineering, Hokkaido University, Sapporo, Japan Abstract. Kabwe area in Zambia has been affected by heavy metal contami-
nations which derived from past mining activities. Particularly, Pb is one of the
most concerned elements for human health in Kabwe. In this context, treatment
by phosphoric fertilizer was conducted to reduce Pb solubility in soil and slag,
limiting their bioavailability. Because leach plant residue in Kabwe contains
metal sulfate minerals with high solubility, concentration of heavy metals in
groundwater is high. We clarified the geochemical behavior of heavy metals
(Pb, Cd, Zn and Cu) after the addition of phosphoric fertilizer (Triple Super
Phosphate: TSP) in column experiment. Immobilization of Pb and Cd lowers
concentration of the metals in ground below WHO environmental standard. Keywords: Insolubilization Soil amendments Heavy metal contamination
Phosphate mineral Mine waste Keywords: Insolubilization Soil amendments Heavy metal contamination
Phosphate mineral Mine waste © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 445–446, 2019.
https://doi.org/10.1007/978-3-030-22974-0_108 References Anderson R (2012) Resource depletion: opportunity or looming catastrophe? https://www.bbc. com/news/business-16391040
Satterthwaite D (2009) The implications of population growth and urbanization for climate
change Environ Urbanization 21(2):545–567 Anderson R (2012) Resource depletion: opportunity or looming catastrophe? https://www.bbc. com/news/business-16391040 Satterthwaite D (2009) The implications of population growth and urbanization for clim
change. Environ Urbanization 21(2):545–567 Satterthwaite D (2009) The implications of population growth and urbanization for climate
change. Environ Urbanization 21(2):545–567 Satterthwaite D (2009) The implications of population growth and urbanization for climate
change. Environ Urbanization 21(2):545–567 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches We conducted a series of column experiments in 50 ml of syringe tubes, simulating
treatment for stacked slags at a dumping site in Kabwe. The syringes were filled with
slags obtained from Kabwe site with 10 g of TSP on the top of slag sample. 6 mL of
rain water obtained from the site was added every day, which is consistent with average
daily precipitation rate. Infiltrated water was collected at the bottom of syringe and
analyzed by ICP-AES and ICP-MS. After the column experiments, the slag samples in
the column also investigated to understand geochemical processes occurred during the
experiments by SEM/EDS. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 445–446, 2019. https://doi.org/10.1007/978-3-030-22974-0_108 H. Kamegamori et al. 446 3
Results and Discussion We confirm the reduction in Pb and Cd concentrations in the eluents. Remarkably, the
reduction for Pb concentration is 96%. In the infiltrated slags simultaneously, we
observe the alteration from PbSO4 to (Pb, Ca)5(PO4)3Cl (Fig. 1), which effects to
reduce the mobility of Pb. In contrast, elution of Zn and Cu from the slags are promoted
by the presence of TSP. This is due to lowering pH by TSP, desorbed Zn and Cu from
amorphous and crystalline iron hydroxides. It suggests to supply orthophosphate at
neutral pH range is effective for immobilization of heavy metals in slags. Fig. 1. The alteration from anglesite to pyromorphite Fig. 1. The alteration from anglesite to pyromorphite 4
Conclusions We confirmed the behavior of some heavy metals applied TSP in column scale. From
the results, TSP could immobilize Pb and Cd, however, it promoted elution of Zn and
Cu due to soil acidification. This suggests applying TSP with dolomite to the slag could
be a better remediation method. Acknowledgements. This study is supported by International Collaborative Research Program
(SATREPS): Visualization of Impact of Chronic/Latent Chemical Hazard and Geo-Ecological
Remediation in Zambia. I’m deeply grateful to Dr. Kasama who taught me how to use SEM/EDS
in Center for Electron Nanoscopy, Denmark Technical University. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 447–450, 2019.
https://doi.org/10.1007/978-3-030-22974-0_109 S. Krivovichev1,2(&), S. Yudintsev3, A. Pakhomova4,
and S. Stefanovsky5 S. Krivovichev1,2(&), S. Yudintsev3, A. Pakhomova4,
and S. Stefanovsky5 1 Kola Science Center, Russian Academy of Sciences, Apatity, Russia
krivovichev@admksc.apatity.ru
2 Department of Crystallography, St. Petersburg State University,
St. Petersburg, Russia
3 Institute of Geology of Ore Deposits, Petrography, Mineralogy,
and Geochemistry, Russian Academy of Sciences, Moscow, Russia
4 Deutsches Elektronen-Synchrotron (DESY), Petra III, Hamburg, Germany
5 Frumkin Institute of Physical Chemistry and Electrochemistry,
Russian Academy of Sciences, Moscow, Russia 1 Kola Science Center, Russian Academy of Sciences, Apatity, Russia
krivovichev@admksc.apatity.ru
2 Department of Crystallography, St. Petersburg State University,
St. Petersburg, Russia
3 Institute of Geology of Ore Deposits, Petrography, Mineralogy,
and Geochemistry, Russian Academy of Sciences, Moscow, Russia
4 Deutsches Elektronen-Synchrotron (DESY), Petra III, Hamburg, Germany
5 Frumkin Institute of Physical Chemistry and Electrochemistry,
Russian Academy of Sciences, Moscow, Russia 1 Kola Science Center, Russian Academy of Sciences, Apatity, Russia
krivovichev@admksc.apatity.ru 2 Department of Crystallography, St. Petersburg State University,
St. Petersburg, Russia Abstract. Murataite-pyrochlore titanate ceramics are attractive waste forms
capable to immobilize radioactive waste streams of complex compositions, thus
eliminating the need for further chemical separation. We have investigated
structures of three types of murataite: 3C, -5C, and -8C phases and demonstrate
their polysomatic nature and structural complexity. Structurally simple pyro-
chlore crystallizes first, followed by crystallization of murataite-5C containing
pyrochlore cells surrounded by fragments of Keggin clusters. This phase is
overgrown by murataite-8C containing both murataite and pyrochlore cells. The
crystallization finishes with the formation of murataite-3C, which is the most
stable and less actinide-rich. The microstructure formed via this mechanism
reminds a Russian doll, which creates additional barrier for the actinide leaching
from the pyrochlore core. The high chemical and structural complexity of the
pyrochlore-murataite series is unparalleled in the world of crystalline materials
proposed for the HLRW immobilization, which makes it unique and promising
for further exploration. Keywords: Murataite Pyrochlore Crystal structure Crystallization
Microstructure Radioactive waste Actinides Keywords: Murataite Pyrochlore Crystal structure Crystallization
Microstructure Radioactive waste Actinides 1
Introduction One of the most important tasks for the advanced nuclear cycle is the elaboration of waste
forms capable to immobilize waste streams of complex compositions, thus eliminating
the need for further chemical separation. In this regard, the murataite-pyrochlore titanate 448 S. Krivovichev et al. ceramics attract considerable attention due to their ability to immobilize radioactive
wastes with different and complex chemical compositions, including actinides such as
Pu-238. Over last few years, there has been a renewed interest in their synthesis and
investigations (Maki et al. 2017; Lizin et al. 2018, etc.). g
(
;
,
)
Murataite-(Y) is a complex titanate mineral first discovered in alkali pegmatites in
St. Peters Dome area in Colorado, United States and later found in pegmatites in the
Baikal region in Russia. Its crystal structure (cubic, space group F-43 m, a = 14.886 Å)
was determined by Ercit and Hawthorne (1995) as based upon a framework of corner-
linked a-Keggin clusters hosting a complex metal-oxide substructure. The simplified
formula of natural murataite-(Y) can be written as [8]R6
[6]M112
[5]M24
[4]TX43, where R = Y,
HREE, Na, Ca, Mn, M1 = Ti, Nb, Na, M2 = Zn, Fe, Ti, Na, T = Zn, Si and X = O, F,
OH. The interest in murataite-(Y) was renewed in 1982, when its synthetic analogue
was identified in Synroc-type titanate ceramics with imitators of high-level radioactive
waste at the Savannah River nuclear power plant (Morgan and Ryerson 1982). Laverov
et al. (1998) reported the formation of murataite-type titanate phase in the uranium-
bearing Synroc matrix from the Mayak factory, a radiochemical facility for the
reprocessing of nuclear fuel located in Southern Ural, Russian Federation. It was found
that five volume percent of synthetic murataite accumulate about 40% of the total
amount of uranium present in the sample, which led to follow-up detailed studies of
chemistry and properties of this material. Transmission electron studies allowed
identification of synthetic varieties of murataite with 3 3 3, 5 5 5,
7 7 7 and 8 8 8 fluorite-like cubic supercells, referred in the following as
murataite-3C, -5C, -7C and -8C phases (Laverov et al. 2011). 2
Methods and Approaches We have studied crystal structures of murataite-3C, -5C and -8C using single-crystal
X-ray diffraction analysis on the samples obtained by melting the mixture of oxides in
an electric furnace at 1500 °C with subsequent cooling to the room temperature as
described by Laverov et al. (1998). The details of the experimental procedures used to
obtain structure models have been described in detail in (Krivovichev et al. 2010;
Pakhomova et al. 2013, 2016). 3
Results and Discussion Urusov et al. (2005) proposed that synthetic murataites can be considered as members
of murataite-pyrochlore polysomatic series consisting of different combinations of 2D
modules. The structural determination of murataite-5C reported by Krivovichev et al. (2010) confirmed the assumption about the modular nature of the polysomatic series
and demonstrated that the murataite- and pyrochlore-type modules are not layers but
zero-dimensional blocks (nanoscale clusters), combination of which in a 3-dimensional
space generates at least two different derivative structures, which combine structural
features of both murataite and pyrochlore. In particular, the crystal structure of
murataite-5C can be described as an ordered arrangement of pyrochlore unit cells Murataite-Pyrochlore Ceramics as Complex Matrices 449 immersed into the recombined murataite matrix, i.e. a substructure consisting of
murataite structure elements. immersed into the recombined murataite matrix, i.e. a substructure consisting of
murataite structure elements. The crystal structure of murataite-3C was reported by Pakhomova et al. (2013),
who demonstrated its general identity to natural murataite, with some important
chemical and structural modifications. The modular nature of the murataite-pyrochlore
polysomatic series was discussed by Laverov et al. (2011). The crystal structure of murataite-8C was reported by Pakhomova et al. (2016) as
based upon a three-dimensional octahedral framework that can be described as an
alternation of murataite and pyrochlore modules immersed into transitional substruc-
ture that combine elements of the crystal structures of murataite-3C and pyrochlore. The obtained structural model confirmed the polysomatic nature of the pyrochlore-
murataite series and illuminated the chemical and structural peculiarities of crystal-
lization of the murataite-type titanate ceramic matrices. Table 1. Information-based structural complexity parameters for the synthetic members of the
pyrochlore-murataite polysomatic series
Material
v [atoms] IG [bits/at.] IG,total [bits/cell]
Pyrochlore
22
1.686
37.088
Murataite-5C
336
4.892
1643.840
Murataite-8C 1387
6.558
9096.031
Murataite-3C
71
3.226
229.044 Table 1. Information-based structural complexity parameters for the synthetic members of th
yrochlore-murataite polysomatic series Table 1. Information-based structural complexity parameters for the synthetic members of the
pyrochlore-murataite polysomatic series 4
Conclusions The information-based complexity parameters for the members of the pyrochlore-
murataite are listed in Table 1. Both kinds of information-based complexity parameters
(per atom and per cell) behave in a similar fashion: they are relatively small for the
initial pyrochlore phase, then increase for murataite-5C, reach their maxima for
murataite-8C and decrease for the final murataite-3C phase. This trend is also followed
in the crystallization of murataite-pyrochlore ceramics: structurally simple and actinide-
rich pyrochlore crystallizes first, creating conditions for the saturation of melt with
Keggin clusters, which triggers crystallization of murataite-5C containing pyrochlore
unit cells surrounded by fragments of Keggin clusters. This pyrochlore-rich phase is
overgrown by murataite-8C containing both murataite and pyrochlore unit cells. The
crystallization finishes with the formation of pure Keggin phase murataite-3C, which is
the most stable and less actinide-rich. The microstructure formed during such a crys-
tallization reminds a Russian doll (‘matryoshka’), which creates additional barrier for
the actinide leaching from the pyrochlore (or crichtonite) core. The high chemical and
structural complexity of the members of the pyrochlore-murataite series is unparalleled
in the world of crystalline materials proposed for the high-level radioactive waste
immobilization, which makes it unique and promising for further technological and
scientific exploration. 450 S. Krivovichev et al. Acknowledgements. This work was supported by the President of the Russian Federation grant
for leading scientific schools (grant NSh-3079.2018.5 to SVK). Acknowledgements. This work was supported by the President of the Russian Federation grant
for leading scientific schools (grant NSh-3079.2018.5 to SVK). Acknowledgements. This work was supported by the President of the Russian Federation grant
for leading scientific schools (grant NSh-3079.2018.5 to SVK). © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 451–452, 2019.
https://doi.org/10.1007/978-3-030-22974-0_110 1
Introduction After the accident at the Fukushima Daiichi Nuclear Power Station that occurred due to
the The2011 off the Pacific coast of Tohoku Earthquake, contaminated water with
radioactive nuclides such as cesium (Cs) have been continuously generated. Zeolite have
been used for removing Cs from the contaminated water, and the spent zeolite are cur-
rently planned to be solidified for the safety storage and disposal. Recently, geopolymer is
considered as a new binder for safety disposal of spent zeolite. Geopolymer has an
advantage that primary phases such as potassium almino silicate hydrate (K-A-S-H) may
have property for sorbing radioactive nuclides. However, there are few data about the
adsorption behavior of Cs by K-A-S-H and the Cs leaching during their alteration. Cs Leaching Behavior During Alteration
Process of Calcium Silicate Hydrate
and Potassium Alumino Silicate Hydrate K. Kuroda1(&), K. Toda1, Y. Kobayashi1, T. Sato2, and T. Otake2 1 Graduate School of Engineering, Hokkaido University, Hokkaido, Japan
k7927k@eis.hokudai.ac.jp
2 Faculty of Engineering, Hokkaido University, Hokkaido, Japan Abstract. Zeolite, used to remove Cs from a contaminated water, would be
solidified for the safety disposal. Recently, geopolymer is considered as a new
binder for disposal. Geopolymer has an advantage that primary phases such as
potassium almino silicate hydrate (K-A-S-H) may sorb radioactive nuclides. In
this study, Cs adsorption, co-precipitation and desorption experiment were
conducted, and C-S-H, which is primary phases of cement, were also employed
for experiments for comparison. From these experiments, it is obtained that K-A-
S-H has higher adsorption capacity of Cs than C-S-H. Cs adsorption ratio and co-
precipitation ratio by C-S-H were almost same. Cs is likely sorbed by C-S-H
thoroughly via ion exchanging. The desorption experiment demonstrated that
most Cs was desorbed from C-S-H while 90% of Cs remained in K-A-S-H. Therefore, K-A-S-H has a higher retention capacity than that of C-S-H. Conse-
quently, geopolymer is considered to be a better material in terms of Cs storage. Keywords: Geopolymer K-A-S-H C-S-H Radioactive waste Keywords: Geopolymer K-A-S-H C-S-H Radioactive waste References Ercit TS, Hawthorne FC (1995) Murataite, a UB12 derivative structure with condensed Keggin
molecules. Can Mineral 33:1223–1229 Krivovichev SV, Yudintsev SV, Stefanovsky SV, Organova NI, Karimova OV, Urusov VS
(2010) Murataite-pyrochlore series: a family of complex oxides with nanoscale pyrochlore
clusters. Angew Chem Int Ed 49:9982–9984 Laverov NP, Sobolev IA, Stefanovskii SV, Yudintsev SV, Omel’yanenko BI, Nikonov BS
(1998) Synthetic murataite: a new mineral for actinide immobilization. Dokl Earth Sci
363:1104–1106 Laverov NP, Urusov VS, Krivovichev SV, Pakhomova AS, Stefanovsky SV, Yudintsev SV (2011)
Modular nature of the polysomatic pyrochlore-murataite series. Geol Ore Dep 53:273–294 Lizin AA, Tomilin SV, Poglyad SS, Pryzhevskaya EA, Yudintsev SV, Stefanovsky SV (2018)
Murataite: a matrix for immobilizing waste generated in radiochemical reprocessing of spent
nuclear fuel. J Radioanal Nucl Chem 318:2363–2372 Maki RSS, Morgan PED, Suzuki Y (2017) Synthesis and characterization of a simpler Mn-free,
Fe-rich M3-type murataite. J Alloys Compd 698:99–102 Morgan PED, Ryerson FJ (1982) A “cubic” crystal compound. J Mater Sci Lett 1:351–352 Morgan PED, Ryerson FJ (1982) A “cubic” crystal compoun Pakhomova AS, Krivovichev SV, Yudintsev SV, Stefanovsky SV (2013) Synthetic murataite-
3C, a complex form for long-term immobilization of nuclear waste: Crystal structure and its
comparison with natural analogues. Z Kristallogr 228:151–156 Pakhomova AS, Krivovichev SV, Yudintsev SV, Stefanovsky SV (2016) Polysomatism and
structural complexity: Structure model for Murataite-8C, a complex crystalline matrix for the
immobilization of high-level radioactive waste. Eur J Mineral 28:205–214 Urusov VS, Organova NI, Karimova OV, Yudintsev SV, Stefanovskii SV (2005) Synthetic
“murataites” as modular members of a pyrochlore-murataite polysomatic series. Dokl Earth
Sci 401:319–325 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches In this study, C-S-H, which is primary phases of cement, were also employed for exper-
iments for comparison. In adsorption experiments, powder C-S-H and K-A-S-H were put
into Cs-solution whose concentration is 1.0 mM at 298 K for a week. And in K. Kuroda et al. 452 co-precipitation experiment, the materials to synthesize C-S-H were put. K-A-S-H could
not be conducted co-precipitation experiment because water react with materials during
synthesize. The solid sample after adsorption experiment were investigated in batch test
and flow-through test as desorption experiment of Cs. The period of batch test is 4months
and that of flow-through test is a month. And deionized water was used in both of them. 3
Results and Discussion The adsorption ratio by K-A-S-H is 92%, while the adsorption ratio by C-S-H is 29%. The reason of this is considered that the size of sorption site is based on ionic radius of K
or Ca, and that of Cs is similar to K than Ca. The adsorption ratio and co-precipitation
ratio by C-S-H is almost same. Cs is likely sorbed by C-S-H thoroughly via ion
exchanging, so it may be easy to sorb even after generation. In batch test as desorption
experiment, the reaction between solid and water phase became equilibrium in 1month,
and Cs concentration were almost stable after that. The desorption ratio from C-S-H was
around 20% and from K-A-S-H was around 2%. But in flow through test, C-S-H desorb
almost all of Cs in a day. It is considered that Cs sorption by C-S-H is ion exchange, so it
is easy to leach by ion exchange too. On the other hand, The desorption ratio from K-A-
S-H was almost 1% per day until 1month had past. Cs/Si ratio in each day was constant,
and Si concentration is considered to depend on the dissolution amount of K-A-S-H. It is
considered that Cs concentration also depended on that. From these results, it can be said
that K-A-S-H has higher property to prevent desorption of Cs than C-S-H. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 453–456, 2019.
https://doi.org/10.1007/978-3-030-22974-0_111 FSBI IMGRE, Moscow, Russia
imgre@imgre.ru Abstract. The main types of environmental impact during exploration,
development and mining of mineral deposits are considered. The indicators of
the environmental situation caused by the mining and mineral processing in the
mining regions, as well as the environmental consequences of accumulated
mining and industrial waste are presented. The results of environmental moni-
toring of the Russian industrial cities are demonstrated. Keywords: Environmental safety Mining Man-made waste
Heavy metals Pollution of the ecosystem 4
Conclusions Consequently, K-A-S-H has higher retention capacity than that of C-S-H. These results
show that geopolymer whose matrix is composed of K-A-S-H is considered to be better
in terms of Cs storage. Acknowledgements. This work was supported by MEXT 8桁の認可番号, Long-term perfor-
mance of cement disposal systems for synthetic zeolites and titanates arising from reprocessing of
contaminated water. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. E. Levchenko(&), I. Spiridonov, and D. Klyucharev E. Levchenko(&), I. Spiridonov, and D. Klyucharev FSBI IMGRE, Moscow, Russia
imgre@imgre.ru FSBI IMGRE, Moscow, Russia
imgre@imgre.ru 2
Methods and Approaches Monitoring of the natural environment should be carried out at all stages of the mining
area life, from exploration to mining and further reclamation of disturbed lands and
until the site becomes completely stabilized. The basis of this paper are ecological and geochemical studies, including the
identification of areas of environmental pollution by toxic substances, assessment of
their extent and composition of their pollution; assessment of potential geochemical
endemicity; zoning of the territory according to the pollution level and the degree of
environmental danger. identification of pollution sources; identifying areas of potential
man-made objects; ecological and geochemical monitoring and forecast of the devel-
opment of negative processes; development of recommendations for the rehabilitation
of areas of poor ecological condition; identification of populations with an increased
risk of morbidity. The result of these studies is the compilation of ecological and
geochemical maps portraying the ecological status of the territory. The study of the environmental health is carried out in the following main areas:
mapping of the man-made pollution in soil and snow cover; establishing the charac-
teristics of the response of plants to soil pollution; geochemical studies of ground and
surface water, and stream sediments; analysis of the chemical composition of atmo-
spheric air, precipitation and aerosols, industrial waste materials as sources of envi-
ronmental pollution and objects for the extraction of secondary raw materials;
relationships of environmental pollution and health indicators of the population living
in the pollution hot spots. 1
Introduction Intensive economic development due to the steady progress in science and technology
entails an inevitable increase in the consumption of minerals. In this regard, the
increase in mineral production during the last century, a sharp increase in the mining
activities contributed to the accumulation of mining waste and man-made pollution of
ecosystems. Besides, despite the obvious benefits of mining for the benefit of man, on
the other hand, it is also a powerful source of environmental hazards for biota and
humans (Aleksandrova and Nikolaeva 2015). Many chemical elements contained in waste products, in addition to industrial
value, cause toxic effects on the ecosystem. The mining of mineral deposits leads to a change in the basic physicochemical
properties of the lithosphere, including its main functions, i.e., geodynamic, geo-
physical, resource, and geochemical. The study of changes in the ecosystem’s
parameters during the life period of a mining enterprise is one of the key goals of an
ecological-geochemical assessment. The high level of the environmental impact is typical of the waste produced by ore
processing and metallurgical operations, since their storage requires special engineering
structures, and the waste contains chemical components harmful to nature and human
health. Their mass is inferior to that of stripped overburden and host rocks, but they
affect the environment more perniciously (Spiridonov and Levchenko 2018). The environmental situation has deteriorated significantly due to the fact that at the
end of the last century after the collapse of the USSR, many large mining complexes
did not cope with economic difficulties and ceased their activities. The tailings of the
enterprises, by majority toxic, have remained uncontrolled. Their conservation and E. Levchenko et al. 454 reclamation have not been carried out timely; hence pollution keeps on growing. In the
soils buried under the dumps tangible geochemical transformations occur. The soils
buried 20 and more years ago display a strong oxidation over the whole depth of their
profile (e.g., pH stays as low as 3.5–4.0), and soil colloids become destroyed. The soil
absorbing complex is disturbed, the mobility of organic matter increases, the soil
horizons gain ore components, which additionally differentiate due to unequal mobility. These facts testify the mobility of chemical elements in the dumps, and the latter often
remain connected to the watercourse systems and can affect the territory of the mining
and processing works in the area of air emissions and waste storages. 3
Results and Discussion The share of mining industries accounts for 70–80% of the volume of all man-made
formations, which have their own characteristics, due to the composition of the feed-
stock, the technology of extraction, enrichment or processing, and a number of other
factors. As demonstrated by ecological and geochemical studies, the most serious negative
effects are related to: the functioning of large industrial hubs (Nizhny Novgorod,
Irkutsk-Cheremkhovo, Khabarovsk, Vladivostok, etc.), as well as exploration and 455 Environmental Pollution Problems development of mineral deposits in active mining areas (Kirovsk, Mama-Bodaibo,
Khapcheranga, Dalnegorsk-Kavalerovsk, Norilsk, and other areas of similar profile). On the basis of the analysis of the updated database of available technogenic
objects, including rare metal deposits, the allocation of 576 technogenic formations on
the territory of the Russian Federation is analyzed. Relevant location maps were compiled, and ranking of technogenic deposits and
formations was carried out using the following parameters: areal extent, storage type(s),
type(s) of technogenic formations, hazardousness classes, and level of environmental
impact. The man-made deposits and formations were ranked by their effect on the
elements of the environment. The analysis of the hottest spots suggests that a series of the factors provokes the
deterioration of the ecological situation in the territories. Of particular concern is the ore processing plant waste, since it requires special
engineering structures, and the waste itself contains chemical elements and compounds
harmful to nature and human health. Their amounts are inferior relatively to the masses
of stripped barren overburden and hosting rocks, but they affect the ecological situation
more perniciously. For example, the environmental situation caused by the extraction
of mineral raw materials and the disposal of waste on more than 25% of the territory of
the Urals economic region is estimated as a crisis. Slightly less than the area of such
lands in the south of the Russian Far East, Khanty-Mansi Autonomous Area, Tyumen
Region, Krasnoyarsk Territory and other areas of intensive mining and processing of
mineral resources. According to the environment impact degree, the highly hazardous objects list is as
follows: apatite concentrates of the Khibiny apatite-nepheline deposits (TR, Sr, F),
enrichment tails of the eudialyte lujavrites of the Lovozero GOK (TR, Th), tailings of
enrichment of baddeleyite-apatite-magnetite ores of the Kovdor Mining apatite, bad-
deleyite ZrO2). Medium-level objects are waste storages accumulated from the apatite
concentrate processing in the Khibiny group deposits (phosphogypsum) containing rare
earth metals and gypsum (Bykhovskiy et al. 3
Results and Discussion 2016; Karnachev et al. 2011). The toxicity of mining products depends on their physical condition and chemis-
tries. Understanding the mechanisms of the action of chemical elements and com-
pounds on the environment and public health makes it possible to optimize medical
consequences and to carry out acceptable mining and processing of mineral raw
materials. At the same time, it is necessary to take into account the whole range of
sources and objects of impact in order to create a system of medical and environmental
safety of the work areas. The problems of the urbanized environment as a human habitat become similar to
those experienced by geologists, representatives of related professions and the popu-
lation of geological exploration, mining, oil and gas, and metallurgical enterprises. Three indicators are accepted in Russia as measures of the soil chemical pollution
in Russia; these are the maximum acceptable content (MAC), the background content
(Zb) and crustal abundance/clarke (Zc). We analyzed the weighted average bulk content
distribution of heavy metals (the hazardousness classes 1 and 2): Pb, Cd, Hg, Zn, Ni,
and Cu. By the above mentioned three evaluation criteria, the cities falling into the 1st
(highly dangerous) category are Irkutsk, Penza, Saratov, Chelyabinsk, Yekaterinburg, E. Levchenko et al. 456 the 2nd (dangerous) are Perm and St. Petersburg, and the 3rd (moderately dangerous)
include Blagoveshchensk and Vologda. the 2nd (dangerous) are Perm and St. Petersburg, and the 3rd (moderately dangerous)
include Blagoveshchensk and Vologda. 4
Conclusions The extent of the loss of land, water, forest, recreational and other resources from
subsoil use in general and from unused waste in particular places these processes on a
par with negative factors that pose a threat to the country’s security. The environmental consequences of accumulated mining and industrial waste are
larger than it is declared in various publications concerning the problem under con-
sideration and are of a global scale. 1
Introduction The growth of industrial and mining production, the progress of civilization increase
environmental problems due to the increasing consumption of mineral and other
resources from the bowels of the Earth, due to the rapid rise in the number of solid
man-made wastes of different productions. These wastes can be used for the production
of building materials and to improve the environmental safety of mining regions. Abandoned lands are exempted from waste dumps and territories have environmental
and economic benefit (Oreshkin 2017). © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 457–460, 2019.
https://doi.org/10.1007/978-3-030-22974-0_112 Environmental Solutions for the Disposal
of Fine White Marble Waste I. Shadrunova, T. Chekushina(&), and A. Proshlyakov Academic N.V. Melnikov Institute of Problems of Comprehensive
Exploitation of Mineral Resources, Russian Academy of Sciences,
Moscow, Russia
tvche.2016@gmail.com Abstract. The article deals with environmental problems of formation of fine
white marble wastes on the territory of Koelga deposit and total mining com-
plex. An inventory analysis of marble waste was carried out, environmental
assessment of fine marble waste and their impact on the ecology of the complex
territory was carried out and theoretically justified. Planned and scientifically
justified ways of large-scale utilization in the production of ceramic bricks. Keywords: Formation of fine marble waste Environmental assessment
Amount of waste Waste disposal Keywords: Formation of fine marble waste Environmental assessment
Amount of waste Waste disposal References Aleksandrova TN, Nikolaeva NV (2015) Ecological-geochemical estimate of the Russian mining
and metallurgy waste. Polytech University Publishers, St. Petersburg (in Russian) Bykhovskiy LZ, Potanin SD, Kotelnikov EI, Anufrieva SI et al (2016) Rare earths and Sc-
bearing man-made formations and deposits in Russia. In: Rare earth and Sc minerals in
Russia: Mineral commodities, VIMS Economic Geology Series, No. 31, pp 112–120 (in
Russian) Karnachev IP, Zhirov VK et al (2011) Ecological and sanitary estimate of the Khibiny mining
area, Murmansk oblast. Vestnik MGU, vol 14, no 3, pp 552–560 (in Russian, with English
abstract) Karnachev IP, Zhirov VK et al (2011) Ecological and sanitary estimate of the Khibiny mining
area, Murmansk oblast. Vestnik MGU, vol 14, no 3, pp 552–560 (in Russian, with English
abstract) Spiridonov IG, Levchenko EN (2018) Mining waste and ecological safety. Prospect and
protection of mineral resources, no 10, pp 15–24 (in Russian, with English abstract) Spiridonov IG, Levchenko EN (2018) Mining waste and ecological safety. Prospect and
protection of mineral resources, no 10, pp 15–24 (in Russian, with English abstract) Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. I. Shadrunova et al. Almost all kinds of new productions require new construction, materials and
mineral resources. Therefore, to improve the environmental safety of the regions it is
necessary to carry out comprehensive development of deposits, and also to utilize man-
made wastes in the production of building materials. To solve the above problems a
comprehensive environmental assessment of man-made waste requires. The assessment
should include amount of accumulated volumes for large-scale utilization of man-made
waste in the production of building materials, products (Khokhryakov et al. 2013). The purpose of the article is the ecological assessment of formation of man-made
waste products of mining production in the form of white marble with a decrease in the
available subsoil mineral resources for the production of building materials and
products. To achieve the goal, it is necessary to justify the use of these wastes as raw
materials for the production of building materials and products. This will simultane-
ously improve the environmental safety of the territories due to large-scale utilization
of man-made waste marble and will free up the areas occupied by dumps. 2
Methods and Approaches In the world and in the Russian Federation there are the technologies of extraction and
processing of non-metallic mineral resources. During these technological processes
man-made wastes are formed. Their queries and the surrounding areas are withdrawn
from economic circulation, violate natural landscapes - their man-made options are
created. It destroys the soil, changes modes of rivers, lakes, reservoirs, underground
and surface groundwater and causes great damage to the environment (Meshheryakov
et al. 2009). Koelga deposit of white marble began the work from 1924. During this time in the
dumps huge amounts of fine wastes of extraction and processing marble were accu-
mulated (Tseytlin 2012). 458 3
Results and Discussion It was calculated that in 2018 the total mass in the dumps is more than 25 million tons
of fine marble wastes, and the area of dumps - more than 20 hectares. An important task
was also the calculation of the environmental damage from the abandoned territories
under the dumps of fine marble waste. To calculate it was analyzed the environmental effect from their utilization by
reducing the area under the dumps, and the pollution of the environment of mining
complex territory (Fadeichev et al. 2012). When calculating it was determined that for the Chelyabinsk region the damage to
the environment from storage of fine marble waste in dumps is about 500 thousand
rubles a year (in the prices of 2018). Taking into account the amount of wastes already
placed in dumps of JSC “Koelgamramor”, the environmental damage will amount to
over 30 million rubles. According to calculations the utilization of fine marble waste in brick production
will significantly reduce the environmental load on the environment. It will take place
by reducing waste mass in dumps, that will allow to reduce the abandoned areas under
dumps and to return the land to use. 3 In the articles it was determined the amount of recyclable fine marble waste at 1 m3
of molding mixture for the production of ceramic bricks of multiple colors: terracotta or
dark brown; light red or pinkish; fawn or straw. The analysis of the results of technical tests showed that, on the basis of fine marble
wastes, it is possible to obtain ceramic bricks of danger class 4, which corresponds to
state standard GOST of the Russian Federation. It was found that burning of over-
moulded ceramic raw makes at temperature 850… 900 °C. It is proved that at that
temperature, the particles of marble are not affected by the process of decarbonization. Therefore, there is no greenhouse gas emissions - carbon dioxide, i.e. ecology of this
mining territory is not the subject to harmful effects. Moreover, as above stated, the
level of danger of marble wastes was higher (class 3) by one step, than the level of 459 Environmental Solutions for the Disposal of Fine White Marble Waste danger of the ceramic bricks produced (4 class) on the basis of these wastes. Also
amounts of energy for manufacture of these ceramic bricks reduced significantly as
compared to common ceramic brick (Moumouni et al. 2016). 4
Conclusions Thus, the total mass accumulated fine marble wastes and environmental damage to
mining area of Koelgo deposit were determined. The technology of improving the
ecology of the region due to large-scale utilization of the above marble wastes in the
production of ceramic bricks was elaborated. The possible number of bricks of different
colors at full disposal of accumulated marble wastes was determined. Using environ-
mental life cycle assessment of finished products based on fine marble wastes the
possibility of obtaining an environmentally safe effective bricks was theoretically
justified and the technology of their production was elaborated. The dependence of
color products from fine marble waste was defined. So, at an amount of 20% of fine
marble waste in the mixture by mass of clay rocks the ceramic brick has a dark brown
color, and at 40% - has straw color. The influence of the elemental composition of the
mixture on the color of the brick was determined (Merem et al. 2017). It was proved that the most environmentally safe, resource-saving way of man-
made waste disposal is their utilization in the production of building materials and
products. This method releases territories abandoned for storing waste and provides
environmental and economic effects from the elimination of dumps (Bilgin et al. 2012). Thus, the environmental problems of the Russian Federation connected with rise of
man-made waste of white marble with a decrease in available reserves of mineral
resources for the production of building materials and products were specified. The
scientific foundations of the integrated environmental assessment methodology of man-
made wastes and their large-scale utilization in the production of building materials and
products were elaborated (Hebhoub et al. 2011). The possibility to use these wastes as
raw components for their production, while solving environmental problems of the
territories due to large-scale utilization of man-made waste was justified. This extends
the raw material base and contributes to the integrated development of bowels, their
mineral and man-made resources. Utilization of man-made waste allows to get a huge
environmental and economic effects on the territory of the Russian Federation. 3
Results and Discussion danger of the ceramic bricks produced (4 class) on the basis of these wastes. Also
amounts of energy for manufacture of these ceramic bricks reduced significantly as
compared to common ceramic brick (Moumouni et al. 2016). Hebhoub H, Aoun H, Belachia M, Houari H Ghorbel E (2011) Use of waste marble aggregates in
concrete. Constr Build Mater 25(3):1167–1171 Fadeichev AF, Khokhryakov AV, Grevcev NV, Cejtlin EM (2012) Dynamics of negative impact
on the environment at different stages of mining development. News High Educ Inst
Mountain Mag 1:39–46 Bilgin N, Yeprem HA, Arslan S, Bilgin A, Günay E, Mars MO (2012) Use of waste marble
powder in brick industry. Constr Build Mater 29:449–457
Fadeichev AF, Khokhryakov AV, Grevcev NV, Cejtlin EM (2012) Dynamics of negative impact
on the environment at different stages of mining development. News High Educ Inst
Mountain Mag 1:39–46
Hebhoub H, Aoun H, Belachia M, Houari H Ghorbel E (2011) Use of waste marble aggregates in
concrete Constr Build Mater 25(3):1167–1171 Bilgin N, Yeprem HA, Arslan S, Bilgin A, Günay E, Mars MO (2012) Use of waste marble
powder in brick industry. Constr Build Mater 29:449–457 © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 461–462, 2019.
https://doi.org/10.1007/978-3-030-22974-0_113 References Bilgin N, Yeprem HA, Arslan S, Bilgin A, Günay E, Mars MO (2012) Use of waste marble
powder in brick industry. Constr Build Mater 29:449–457 Fadeichev AF, Khokhryakov AV, Grevcev NV, Cejtlin EM (2012) Dynamics of negative impact
on the environment at different stages of mining development. News High Educ Inst
Mountain Mag 1:39–46 Hebhoub H, Aoun H, Belachia M, Houari H Ghorbel E (2011) Use of waste marble aggregates in
concrete. Constr Build Mater 25(3):1167–1171 I. Shadrunova et al. 460 Khokhryakov AV, Fadeichev AF, Cejtlin EM (2013) Application of an integral criterion for
determining the environmental hazard of mining enterprises. News Ural State Mining Univ
1:25–31 Merem EC et al (2017) Assessing the ecological effects of mining in West Africa: the case of
Nigeria. Int J Mining Eng Mineral Process 6(1):1–19 Meshheryakov YuG, Kolev NA, Fedorov CV, Suchkov VP (2009) Stroymaterialy Production of
granulated phosphogypsum for the cement industry and building products, vol 5, pp 104–106 Moumouni A, Goki NG, Chaanda MS (2016) Natural Resources Geological exploration of
marble deposits in Toto Area, Nasarawa State, Nigeria, vol 7, pp 83–92 Oreshkin DV (2017) StroymaterialyInvironmental problems of integrated development of
mineral resources in the large-scale utilization of man-made mineral resources and waste in
the production of building materials, vol 8, pp 55–63 Tseytlin EM (2012) Features of environmental hazard assessment of mining enterprises Theses of
the report of VII Krakow conference of young scientists. AGH University of Science and
Technology, Krakow, pp 809–819 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction The use of new reserves, in economically viable conditions depends on the available
technologies. The development of deep and ultra-deep offshores requires considerable
research and development efforts. Progress has also been made in managing the
multiple risks associated with this activity. A disaster like «Deepwater Horizon» led to
a detailed analysis and sharing of findings by industry experts. Despite the security rules on the platforms, in fact, some major incidents are
revealed, the causes of which are multifactorial in nature and which have dire conse-
quences for both humans and the environment. Legislation forces organizations to take responsibility for dealing with disasters,
which has been developed over time and in different ways in different countries. Our analysis of how environmental risks are taken into account by various subjects
and offers development prospects to ensure better safety of offshore activities. Security Test of New Technology in View
of Increased Performance of Oil Platforms
Without Increasing Environmental Risks E. M. Tanoh Boguy(&) and T. Chekushina
Department of Mineral Developing and Oil & Gas Engineering,
Engineering Academy, RUDN University, Moscow, Russia
boguymartialeddy@gmail.com E. M. Tanoh Boguy(&) and T. Chekushina Department of Mineral Developing and Oil & Gas Engineering,
Engineering Academy, RUDN University, Moscow, Russia
boguymartialeddy@gmail.com Abstract. In this article, it will be important to note the context of the gradual
depletion of existing fields, which are pushing to expand research and
exploitation of new fossil fuel resources in order to meet the growing demand for
fuel and, despite international regulations to combat global warming. Conse-
quently, an increase in offshore platforms in global hydrocarbon production to
compensate for the depletion of the earth’s reserves is becoming a major problem
for the oil industry. Given the financial unforeseenness that is represented, and
the energy autonomy which is provided, marine exploitation has become a
problem for states with a large sea area and, therefore, an environmental. Keywords: Security test New technology Increased performance
Environmental risks Keywords: Security test New technology Increased performance
Environmental risks Keywords: Security test New technology Increased performance
Environmental risks 2
Methods and Approaches Some of the accidents at oil rigs, such as the Deepwater Horizon, in the spring of 2010
caused a shock wave in their magnitude and severity that convinced that such accidents
could occur. In fact, some states have taken steps to raise the level of security. In fact, E. M. Tanoh Boguy and T. Chekushina 462 some states have taken upon themselves the task of “solving the problem of providing
security on the shelf.” Global hydrocarbon production is becoming more and more
offshore, accounting for more than 35% of oil and 19% of gas. Since the deposits are
located at great depths, states and companies must develop the potential for their use,
seeking to control the risks inherent in this activity carried out under extreme condi-
tions. In order not to have restrictions in the conditions in which exploration, drilling
and mining operations are carried out, by more and more complex and risky methods. In addition, oil companies are well aware of their interest in investing in the devel-
opment of new technologies. This allows them to gain an industrial competitive
advantage in strategic areas of deepwater exploitation. Significant progress has been
made in managing the multiple risks inherent in offshore operations. Oil companies put
prevention at the level of operating conditions. 3
Results and Discussion Regardless of the achievements observed, it is obvious that the safety rules applied on the
platforms guarantee greater efficiency than environmental protection, and that more and more
risks are encountered. Increasing risks to humans and the environment is inextricably linked
with the complexity of drilling operations. Working platforms continue to find solutions that
completely avoid any potential risks in the protected areas. Advanced technology and security
measures suggest that there is a clear improvement, but the limit between politically correct
and pollution is quickly exceeded when it comes to such profits. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 463–465, 2019.
https://doi.org/10.1007/978-3-030-22974-0_114 1
Introduction Calcite is the main mineral component of travertine/tufa, and it plays a decisive role in
the sedimentary evolution of travertine, whether inorganic or bio-organic (Pentecost
1995). The size of the calcite in travertine is a reflection of the deposition rate and can
therefore be used to characterize its sedimentary environment, which is the result of
physical, chemical and biological synergy during the deposition process. Herein, we
divide the calcite in the Huanglong cold-water travertine into different mineral gen-
erations according to the sedimentary environment and evolution time series, i.e., from
the generation of the parents to the descendants. The classification of these mineral
generations helps to understand the physical, chemical and biological processes within
their formation, and then to reconstruct the sedimentary paleo-environment and paleo-
climate. On the other hand, the mineral generation of calcite will help to understand the
architecture of travertine landscape (Wang et al. 2018), so that they can be better
protected and leave more natural heritage of travertine for human beings. Calcite Mineral Generation in Cold-Water
Travertine Huanglong, China F. Wang, F. Dong(&), X. Zhao, Q. Dai, Q. Li, Y. Luo, and S. Deng School of Environment and Resource,
Southwest University of Science and Technology, Mianyang, China
fqdong@swust.edu.cn Abstract. Mineral generations could help us to understand the physical,
chemical and biological processes within their formation, and then to reconstruct
the sedimentary paleo-environment and paleo-climate. The calcite in the
Huanglong cold-water travertine can be divided into three mineral generations,
which reveal two different sedimentary environment systems respectively. In the
calcium cycle, calcite mineral generation exposes a step in recycling marine
matter to the land, and it also allows the land to proliferate, which mainly
manifeste in the addition of plant debris, algae and microbial residues, so that
the topography has been accumulating. Keywords: Cold-water travertine Mineral generation Paleo-environment
Huanglong 4
Conclusions Finally, it is important to include risk management in determining policies, procedures
and plans, as well as specific risk mitigation measures that will be taken to manage
security risks. The environment is associated with all sorts of accidents, while drilling and
operating the platform. The accident, which is a major problem for these exploitation will
be a hydrocarbon spill, which is highly unlikely and will be limited to pumping oil and
fuel stored on support vessels in the event of a tank failure or reloading pipe. Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 3
Results and Discussion From the diagenetic time series of calcite, the types of rocks are Mesozoic limestone
and dolomite, and the travertine deposited since the Late Cenozoic. The calcite in
travertine is further divided into two mineral generations, namely calcite and secondary
in primary travertine travertine. The calcite in travertine is further divided into two
mineral generations, namely calcite in primary travertine and calcite in secondary
travertine. Therefore, calcite is divided into three generations from the generation of the
parent to the descendants, i.e., calcite in the Mesozoic carbonate rock, calcite in the
Late Cenozoic travertine, and calcite re-precipitated after travertine leaching. The calcite in the Mesozoic limestone is micritic, microcrystal and sparry calcite,
ranging in size from centimeters to micrometers. CaO and MgO in the rocks composed
of these calcite are close to the theoretical value, and the other components are very
low, which belong to the soluble carbonates. These calcite became the parent gener-
ation in the whole calcite evolution sequence, and they provided the material source for
the calcite of later generations after being leached. The calcites of the descendants form
the different morphologies of the cold-water travertine. During the formation process,
physical, chemical, biological and other factors participate in the diagenesis. Among
these travertines, no matter what color, except for calcite, other minerals hardly
develop. The calcites of the descendants of travertine are very numerous and complex. Here, we mainly listed two of them, which are calcite in the laminal travertine and
calcite in the porous travertine, because these are the main components of most tra-
vertines. The calcite of the laminal travertine is long columnar and slablike. The brown
and white calcite is continuously growing without interruption. These characteristics
are very different from those observed on the eye assay, which indicates that the calcite
growth in the dry and cold seasons is continuous (Wang et al. 2014). On the other hand,
it reflects that the hydrodynamic conditions are very stable, and the water layer is very
thin with little or no biological involvement. The calcite in the porous travertine tends
to be granular, and the particle size is much smaller than that of the laminal travertine,
and its particle size is generally less than 100 lm. 2
Methods and Approaches A comprehensive field geological survey of rocks consisting of calcite was performed,
mainly from sedimentary rocks, and systematic sample collection based on the geo-
logical background of these rocks was carried out. The mineralogy studies of calcite F. Wang et al. 464 were carried out by polarized light microscopy, XRD and SEM to determine their
generational relationship. were carried out by polarized light microscopy, XRD and SEM to determine their
generational relationship. Calcite Mineral Generation in Cold-Water Travertine Huanglong, China Calcite Mineral Generation in Cold-Water Travertine Huanglong, China 4
Conclusions The calcite in the Huanglong cold-water travertine can be divided into three mineral
generations, which reveal two different sedimentary environment systems respectively. They are the generations of the marine carbonate rock diagenesis system, and the
descendant generation is the continental freshwater karst sedimentary system. Unlike
conventional weathering, which converts terrestrial carbonate rocks to the ocean phase,
this is done in the opposite direction. The study of different calcite mineral generation
can reconstruct the paleo-environment and paleo-climate of its sedimentation. Acknowledgements. This research was supported by National Natural Science Foundation of
China (Grants nos. 41572035, 41603041 and 41877288), the Open Funds of Key laboratory of
mountain hazards and surface processes (grant No. 19zd310501) and Longshan Talents program
of Southwest University of Science and Technology (18lzx663). 3
Results and Discussion These characteristics reflect the rapid
crystallization of calcite, Due to the strong hydrodynamics and the participation of
biological effects, calcite cannot be continuously grown, but suddenly nucleates and
grows to a certain extent then no longer grows. The last generation of calcite is the secondary calcite in travertine. The ancestral
body of this type of calcite is the deposited travertines, which are dissolved in the water
by weathering and leaching, then the calcite re-precipitates through the deposition of a
parent-like travertine. These calcites will adhere to the cracks, edges and even the
surface of the primary calcite. 465 Pentecost A (1995) The quaternary travertine deposits of Europe and Asia Minor. Quaternary Sci
Rev 14(10):1005–1028
Wang HJ, Yan H, Liu ZH (2014) Contrasts in variations of the carbon and oxygen isotopic
composition of travertines formed in pools and a ramp stream at Huanglong Ravine, China:
implications for paleoclimatic interpretations. Geochimica et Cosmochimica Acta 125:34–48
Wang FD, Dong FQ, Zhao XQ (2018) The large dendritic fissures of travertine dam exposed by
Jiuzhaigou earthquake, Sichuan, southwestern China. Int J Earth Sci 107(8):2785–2786 (
)
q
y
p
p
Q
y
Rev 14(10):1005–1028
Wang HJ, Yan H, Liu ZH (2014) Contrasts in variations of the carbon and oxygen isotopic
composition of travertines formed in pools and a ramp stream at Huanglong Ravine, China:
implications for paleoclimatic interpretations. Geochimica et Cosmochimica Acta 125:34–48
Wang FD, Dong FQ, Zhao XQ (2018) The large dendritic fissures of travertine dam exposed by
Jiuzhaigou earthquake, Sichuan, southwestern China. Int J Earth Sci 107(8):2785–2786 Rev 14(10):1005 1028
Wang HJ, Yan H, Liu ZH (2014) Contrasts in variations of the carbon and oxygen isotopic
composition of travertines formed in pools and a ramp stream at Huanglong Ravine, China:
implications for paleoclimatic interpretations. Geochimica et Cosmochimica Acta 125:34–48
Wang FD, Dong FQ, Zhao XQ (2018) The large dendritic fissures of travertine dam exposed by Pentecost A (1995) The quaternary travertine deposits of Europe and Asia Minor. Quaternary Sci
Rev 14(10):1005–1028 Wang FD, Dong FQ, Zhao XQ (2018) The large dendritic fissures of travertine dam exposed by
Jiuzhaigou earthquake, Sichuan, southwestern China. Int J Earth Sci 107(8):2785–2786 © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 466–470, 2019.
https://doi.org/10.1007/978-3-030-22974-0_115 References Pentecost A (1995) The quaternary travertine deposits of Europe and Asia Minor. Quaternary Sci
Rev 14(10):1005–1028 Wang FD, Dong FQ, Zhao XQ (2018) The large dendritic fissures of travertine dam exposed by
Jiuzhaigou earthquake, Sichuan, southwestern China. Int J Earth Sci 107(8):2785–2786 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Optimization of the Natural-Technical System
“Iron Ore Quarry” Management Based
on the Algorithm of the Rock Mass
Stability Ensuring L. Yarg(&), I. Fomenko, and D. Gorobtsov Department of Engineering Geology,
Russian State Geological Prospecting University (MGRI), Moscow, Russia
ifolga@gmail.com Abstract. The method of natural-technical system (NTS) “Iron ore deposits”
optimal control in terms of the pit walls stability is based on two-level systems
with cross-links. The algorithm for optimizing the pit walls angles designed
values includes the following steps: separation of rock massif into engineering-
geological complexes (EGC), typing of the pit walls within the EGC, substan-
tiation of the calculation geomechanical models and stability analysis of the pit
walls based on mathematical modeling. Based on the results of the calculations
the maximum angle of the pit wall is determined at which it remains stable. As
minimized performance criteria the deviations of the stability factors current
state from the maximum allowable values are considered. The proposed
approach is one of the ways to ensure the stability of the deep-pit quarries walls
during their long-term development. Keywords: Natural-technical system (NTS) Open pit Stability assessment
Optimization of pit walls angles Control of NTS Keywords: Natural-technical system (NTS) Open pit Stability assessment
Optimization of pit walls angles Control of NTS 2
Methods and Approaches The processes development initiated by technological work is progressive in space and
time. Long-term exploration of deposits leads to the changes of boundaries, mode and
set of processes (Bondarik and Yarg 2015). The system of engineering geological support in the quarry areas includes a range
of work and research aimed to obtain the information about engineering geological
conditions during the entire life of the quarry, assessment and forecast of the slope
stability at various stages of their construction to achieve the technical, economic and
environmental safety of mining work. Effective management of the natural-technical system “Iron Ore Deposit KMA”
should be carried out taking into account both local and global stability factors of the
pit walls. Separation of the Rock Massif to EGC. Features of engineering-geological conditions
including lithologic-petrographic composition, physical and mechanical properties,
structural disturbance, parameters of the natural stress field require an individual
approach to the process of predicting the behavior of an array of rocks. This becomes
possible only on the basis of correct engineering and geological research data. Elementary NTS “Stoilensky Quarry” is divided into two engineering-geological
complexes (Yarg et al. 2018): • The upper one is composed of loose and semi-rock soils with a thickness up to
90 m. The sedimentary cover is typified taking into account the geological struc-
ture, hydrogeological conditions (the water inflow along the open-pit contour water
permeability) and the physical and mechanical properties of the soils. • The lower EGC is represented by rocks with a thickness of up to 600 m. The main
stability determining factors are: anisotropy of the massif properties due to its
fracturing and spatial orientation of the cracks. Engineering geological processes developing during the operation of the elemen-
tary NTS “Open pit” of the iron ore deposits of Stoilensky and Lebedinsky GOK are:
scree formation; collapse; landslides; surface erosion; suffusion: mechanical, chemical;
filtration deformations. Tiping of the Pit Walls Within the EGC. A “bowl” of a quarry with a simple structural
plan of the rock mass (Fomenko et al. 2016, Hoek and Bray 1981, Wyllie and Mah
2010) (i.e. assuming that the direction of weak zones and fracturing within the pit
remains constant) can be divided into zones of conditional stability and potential
instability of the walls (Fomenko et al. 2016). In accordance with these factors, three types of pit walls quarrying were identified:
relatively difficult, difficult and very difficult. 1
Introduction The development of iron ore deposits and permanent deepening of the open pit leads to
changes in the stress state, decompaction of rocks, an increase in massif fracture,
weathering rates and a decrease in the strength properties of rocks that form the open
pit, activation of geological processes: debris, rock falls, landslips and landslides,
suffusion, surface erosion. The considered natural-technical system “Ore deposits of KMA” is a complex
system of the local level. The functioning of the local NTS “Iron Ore Quarry” is
characterized by: a certain set of processes developing permanently without the sta-
bilization stage under the influence of long-term man-made interactions which form the
basis of the NTS operation. Reduction of negative consequences is possible only with a
clear understanding of the processes developing in the field of interaction of natural-
technical systems (NTS) “mining and processing plant (GOK”) (Yarg et al. 2018). Research objective: optimization of the NTS “iron ore quarry” management based
on the algorithm of the rock mass stability ensuring. 467 Optimization of the NTS “Iron Ore Quarry” Management 3
Results and Discussion Optimization of the NTS “Mineral Deposit” functioning is based on a modern
methodology for stability calculation (Pendin and Fomenko 2015, Bar et al. 2018). 468 L. Yarg et al. For potentially unstable pit wall the probable collapse can occur according to the
following schemes: For potentially unstable pit wall the probable collapse can occur according to the
following schemes: 1. The azimuth of crack systems fall coincides with the azimuth of the pit wall fall. In
this case a flat problem can be solved. 1. The azimuth of crack systems fall coincides with the azimuth of the pit wall fall. In
this case a flat problem can be solved. 2. The azimuth of crack systems fall does not coincide with the azimuth of the pit wall
fall, but at the same time according to the kinematic analysis results the formation of
wedge-type collapses is likely. In this case the pit wall stability problem is solved in
a three-dimensional formulation, for example using the method of volume blocks. Based on the results of the calculations the maximum pit wall angle is determined at
which it remains stable. In accordance with the “large-scale interconnected” theory (Tsurkov and Lit-
vinchev 1994), the management of local NTS “Iron Ore Quarry” in terms of the pit
walls stability can be based on two-level systems with cross-links. As minimized
performance criteria the deviations of the stability factors current state from the
maximum allowable values are considered. As optimized parameters the following
were taken: the level of the upper Jurassic aquifer, the strength properties of Alb-
Cenomanian sands and Devonian clays, fracturing and blockiness of the Precambrian
massif. The graphs (Figs. 1, 2 and 3) of the relation between safety factor and the dynamics
of aquifer, blockiness and strength properties of rocks allow setting the limit values of
the system coordinates at which the system does not leave the zone of admissible states. 1.19
1.29
0
5
10
15
Fig. 1. The effect of groundwater level rise (horizontal axis) on the global safety factor of the pit
walls (vertical axis). 1.19
1.29
0
5
10
15 Fig. 1. The effect of groundwater level rise (horizontal axis) on the global safety factor of the pit
walls (vertical axis). 1.1
1.2
1.3
0
5
10
15
20
25
Fig. 2. The relation between Ku (vertical axis) and the blockiness of the rock massif (distance
between cracks), (horizontal axis) 1.1
1.2
1.3
0
5
10
15
20
25 Fig. 2. 4
Conclusions The obtained results are in good agreement with the generally accepted ideas about the
causes of stability infraction in the massif zones near the pit walls. At the same time the
obtained data advantage is the possibility of establishing the limiting values of changes
in the safety factors on graphs. These safety factors determine both local and global
stability of the pit walls at which the system will not leave the zone of admissible states. Control solutions that ensure the safe work performance should include: adjustment
of the drainage system and water supply process, taking into account the position of the
GWL; interception and organized disposal of surface and seepage water into the cir-
culating system; maintaining the water level in the settling ponds, excluding flooding of
the territory; adjustment of the blasting operations technology during the quarrying of
deep horizons taking into account the stress state of the massif and the occurrence of
the rock technogenic fracturing during drilling and blasting operations. g
g
g
g
g
p
The obtained predictive estimates of the pit walls stability can be used in the design
and development of fields with similar engineering geological conditions. L. Yarg et al. The relation between Ku (vertical axis) and the blockiness of the rock massif (distance
between cracks), (horizontal axis) Optimization of the NTS “Iron Ore Quarry” Management 469 1.2
1.25
1.3
1
2
3
Fig. 3. The effect of the ore-crystalline rocks strength (horizontal axis) in the massif on the
safety factor (vertical axis): 1 – densely fractured rocks; adhesion C = 690 kPa, angle of internal
friction u = 32°; 2 ─moderately fractured, adhesion C = 1130 kPa, angle of internal friction
u = 36°; 3 ─weakly fractured, adhesion C = 3140 kPa, angle of internal friction u = 39° 1.2
1.25
1.3
1
2
3 Fig. 3. The effect of the ore-crystalline rocks strength (horizontal axis) in the massif on the
safety factor (vertical axis): 1 – densely fractured rocks; adhesion C = 690 kPa, angle of internal
friction u = 32°; 2 ─moderately fractured, adhesion C = 1130 kPa, angle of internal friction
u = 36°; 3 ─weakly fractured, adhesion C = 3140 kPa, angle of internal friction u = 39° References Bar N, Weekes G, Welideniya S (2018). Benefits and limitations of applying directional shear
strengths in 2D and 3D limit equilibrium models to predict slope stability in highly
anisotropic rock masses. https://www.researchgate.net/profile/Neil_Bar/research Bar N, Weekes G, Welideniya S (2018). Benefits and limitations of applying directional shear
strengths in 2D and 3D limit equilibrium models to predict slope stability in highly
anisotropic rock masses. https://www.researchgate.net/profile/Neil_Bar/research Bondarik GK, Yarg LA (2015) Engineering geology. Questions of theory and practice. Philosophical and methodological foundations of geology. KDU (in Russian) Bondarik GK, Yarg LA (2015) Engineering geology. Questions of theory and practice. Philosophical and methodological foundations of geology. KDU (in Russian) Fomenko IK, Pendin VV, Gorobtsov DN (2016) Estimation of the stability of quarries of quarries
in rocky soils. Min Sci Technol (3), 10–21 (in Russian) Fomenko IK, Pendin VV, Gorobtsov DN (2016) Estimation of the stability of quarries of quarries
in rocky soils. Min Sci Technol (3), 10–21 (in Russian) Hoek E, Bray JW (1981) Rock slope engineering, 3rd edn. Institution of Mining and Metallurgy,
London Hoek E, Bray JW (1981) Rock slope engineering, 3rd edn. Institution of Mining and Metallurgy,
London Pendin VV, Fomenko IK (2015) Methodology of landslide hazard assessment and forecast. Publishing House of the Russian Federation Lenand, Moscow (in Russian) Pendin VV, Fomenko IK (2015) Methodology of landslide hazard assessment and forecast. Publishing House of the Russian Federation Lenand, Moscow (in Russian) L. Yarg et al. 470 Tsurkov VI, Litvinchev IS (1994) Decomposition in dynamic problems with cross-links. Science:
Physics and Mathematics, Moscow (in Russian) Yarg LA, Fomenko IK, Zhitinskaya OM (2018) Evaluation of slope optimization factors for
long-term operating open pit mines (in terms of the Stoilensky iron ore deposit of the Kursk
Magnetic Anomaly). Gornyi Zhurnal (11), 76–81 Yarg LA, Fomenko IK, Zhitinskaya OM (2018) Evaluation of slope optimization factors for
long-term operating open pit mines (in terms of the Stoilensky iron ore deposit of the Kursk
Magnetic Anomaly). Gornyi Zhurnal (11), 76–81 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. Tsurkov VI, Litvinchev IS (1994) Decomposition in dynamic problems with cross-links. Science:
Physics and Mathematics, Moscow (in Russian) Tsurkov VI, Litvinchev IS (1994) Decomposition in dynamic problems with cross-links. Science:
Physics and Mathematics, Moscow (in Russian)
Wyllie DC, Mah CW (2010) Rock slope engineering: civil and mining, 4rd edn. Spon
Press/Taylor&Francis Group, London
Yarg LA, Fomenko IK, Zhitinskaya OM (2018) Evaluation of slope optimization factors for
long-term operating open pit mines (in terms of the Stoilensky iron ore deposit of the Kursk
Magnetic Anomaly). Gornyi Zhurnal (11), 76–81 Yarg LA, Fomenko IK, Zhitinskaya OM (2018) Evaluation of slope optimization factors for
long-term operating open pit mines (in terms of the Stoilensky iron ore deposit of the Kursk
Magnetic Anomaly). Gornyi Zhurnal (11), 76–81 Wyllie DC, Mah CW (2010) Rock slope engineering: civil and mining, 4rd edn. Spon
Press/Taylor&Francis Group, London References The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction Associated petroleum gas (after this APG) is a mixture of various gaseous hydrocar-
bons dissolved in oil and released in the process of extraction and preparation of oil. The oil gases also include gases released in the operations of thermal processing of oil
(cracking, reforming, hydrotreating, etc.), consisting of saturated and unsaturated
(methane, ethylene) hydrocarbons. From geology, APG is often formed during the Ordovician and Silurian periods. Sometimes it is built late to the Cretaceous. (Vorobiev and Zhang 2018). Utilization of Associated Oil Gas:
Geo-ecological Problems and Modernization
of the State L. Z. Zhang1,2 and H. Y. Sun1,3(&) L. Z. Zhang1,2 and H. Y. Sun1,3(&) 1 Department of Mineral Developing and Oil & Gas Engineering,
RUDN University, Moscow, Russia
657273629@qq.com
2 Liaoning Shihua University, Fushun, China
3 Qinhuangdao Experimental Middle School, Qinhuangdao, China Abstract. In the world vast of oil is extracted, especially in China. Respec-
tively produce associated petroleum gas is in a large volume. There are geo-
ecological problems in the utilization of associated petroleum gas. In connection
with the increasing requirements for the preservation of the state of the bio-
sphere in China, the process of modernization was begun. Chinese modern-
ization of associated petroleum gas utilization is presented. Keywords: Associated petroleum gas Modernization Technology
Geo-ecological problems © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 471–472, 2019.
https://doi.org/10.1007/978-3-030-22974-0_116 2
Methods and Approaches The PRC’s world ranking in oil production is quite high and, accordingly, the volumes
of simultaneously produced associated gas are very significant. In 2011–2013 in China,
APG was provided in the amount of 27.3, 28.9, 30.2 billion m3. In addition, the share
of production took more than 5.1% in the world. Previously, in China, APG was traditionally considered not as a valuable resource,
but as a by-product of oil production, the simplest method of which utilization is flaring
in many fields, especially in Northeast China. 472 L. Z. Zhang and H. Y. Sun 4
Conclusions Associated petroleum gas will become a valuable raw material for further processing. China’s economy needs to use APG to reduce greenhouse gas emissions. Modernization of processing of APG in China and its prospects lie in the area of
increasing the efficiency of processing of associated gas, reducing energy consumption
in the course of processing, flexible operation, convenient installation and operation. 3
Results and Discussion Associated gas recovery technology using membrane separation is based on the fol-
lowing steps: – removing micro solid particles, crude oil and heavy hydrocarbon emulsion con-
tained in associated gas; – removing micro solid particles, crude oil and heavy hydrocarbon emulsion con-
tained in associated gas; – after preliminary impurity removal, heating up to 590 °C in a heat exchanger;
introd cing into a liq id rotar
compressor; – after preliminary impurity removal, heating up to 590 °C in a heat exchanger;
– introducing into a liquid rotary compressor; – introducing the heated gas into a desulfurization tank, and desulfurizing; – introducing the desulfurized gas into a membrane separator, and separating; – introducing the gas from the membrane separator to a molecular sieve tank, and
performing deep desulfurization and decarburization; – cooling to obtain the product (Mo 2013). This technological process is quite simple and convenient in industrial operation,
and besides, it is characterized by low operating costs, high recovery rate and can be,
after a little adaptation, widely circulated. References Mo JL (2013) Recovery process for petroleum associated gas. China Patent CN102994180A, 27
March 2013
Vorobiev AE, Zhang LZ (2018) Apply innovative technologies for processing of associated gas
in China. Eurasian Sci. J. 10(2) Mo JL (2013) Recovery process for petroleum associated gas. China Patent CN102994180A, 27
March 2013
Vorobiev AE, Zhang LZ (2018) Apply innovative technologies for processing of associated gas
in China. Eurasian Sci. J. 10(2) Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Identifying the Decorative Stone Samples
from the Mining Museum’s Collection:
First Results N. Borovkova1(&) and M. Machevariani2 N. Borovkova1(&) and M. Machevariani2 1 Mining Museum, St. Petersburg Mining University, St. Petersburg, Russia
borovkova_nv@pers.spmi.ru
2 Assistant of the Department of Mineralogy, Crystallography and Petrography,
St. Petersburg Mining University, St. Petersburg, Russia Abstract. The report presents the primary results of a study of a unique col-
lection of polished decorative stone samples belonging to Empress Catherine the
Great. Primary macroscopic analysis of 83 plates, divided into 13 groups
according to similar features, was performed. The bulk chemical composition of
rocks was estimated on the basis of XRF- analysis data, performed using a Delta
Olympus XRF portable analyzer. Preliminary studies allowed to outline the
characteristic fields of the studied samples of decorative rocks on the ternary
plots of their bulk composition. In the future, it is planned to perform Raman
spectral imaging to generate detailed maps of the mineralogical composition of
the decorative stone samples. Keywords: Polished decorative stones
Collections of Empress Catherine the great Handheld XRF analyzer
Mining museum Keywords: Polished decorative stones
Collections of Empress Catherine the great Handheld XRF analyzer
Mining museum Keywords: Polished decorative stones
Collections of Empress Catherine the great Handheld XRF analyzer
Mining museum 1
Introduction Natural stone serves as a unique raw material for the objects of decorative and applied
arts, as well as architectural monuments. Museums and monuments of St. Petersburg
store rare objects made of natural stone. The problems of their preservation are
increasingly forcing restorers and art researchers to turn to geologists to identify var-
ious types of gemstone materials needed for restoration. This raises a number of
problems, primarily related to the lack of reliable information about the origin of
various types of such materials, as well as their accurate identification in art objects. Their study is complicated by the need to use exclusively non-destructive analytical
techniques, which greatly complicates the task. Unlike European museums, in Russia,
there is no complex reference collection of natural decorative (ornamental) stone,
supported by current results of laboratory research, reliable information on the location,
and a complete catalog of art object made of such materials. Obviously, the need for
such data is highly in demand not only among restorers and art historians but also
among geologists, whose research interests include the preservation of the diversity of
gemstone raw materials and objects of cultural heritage. © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 475–478, 2019. https://doi.org/10.1007/978-3-030-22974-0_117 476 N. Borovkova and M. Machevariani The object of this study was one of the oldest collections of polished flat samples of
natural decorative (ornamental) stone of the XVIII century, previously owned by
Empress Catherine the Great (Borovkova 2017). The research contributes to the
development of a methodology for evaluating historical gemstone materials of con-
siderable cultural, museum and scientific value. In 1816, a collection was transferred
from the Imperial Hermitage to the Museum of the Mining Cadet Corps (now the
Mining Museum), which included collections of marbles and «rock sampled as pol-
ished plates». In the inventory of this collection, which is stored in the archives of the
State Hermitage Museum, there is the following note: «Jasper and solid rocks found in
the Ural Mountains, starting from Tura River by noon over the rivers Uyu and Ural»
[State Hermitage archives. F. 1. O. 6l. D. 1 a-c]. The total number of such samples is
not indicated, but in the catalog (1798) they are recorded from №1525 to №1725; and
in the next catalog (1811), there are a number of additional samples. 1
Introduction Thus, with the
same description, samples from №2052 to 2330 are recorded in a later catalog. 2
Methods and Approaches Currently, more than 100 items of such decorative rocks are found in the Mining
Museum. The primary macroscopic evaluation of the samples suggested wider geog-
raphy of their origin. The need for reliable authentication of historical samples
necessitated their thorough review. As the first phase of the study, 83 plates were selected and broadly classified into 13
groups. The groups were formed on the basis of visual estimation of similar charac-
teristics: the rocks structure, texture, and color. Eight of the thirteen selected groups
were pre-diagnosed as known geo-referenced decorative rock types that received code
names corresponding to their regional and historical affiliation, namely: Korgon por-
phyry; Tigiretsky breccia; Tigiretsky quartz; Kalkan, Kushkuldinskaya, Nikolaevskaya,
Urazovskaya, and Surguchnaya jasper. The remaining groups require further diag-
nostics and are pre-defined as andesite, hornfels, marble, and green marble. The planned research method involves the use of known geo-referenced samples. Comparative analysis of the studied samples and reference rocks will be divided into
three steps: visual comparison, comparison of chemical and mineral compositions. As mentioned above, the study of museum objects requires a special approach and
the use of non-destructive techniques. In this regard, the samples chemical composition
analysis was performed using a Delta Olympus XRF handheld analyzer. The mea-
surements were carried out in the Mining mode with preliminary calibration. There was
threefold spectra collection from each point with a 30 s acquisition time. In general, 50
bulk chemical composition analysis of rocks belonging to seven previously selected
groups (Korgon porphyry, Tigiretsky quartz, Urazovskaya jasper, green marble, mar-
ble, andesite, and hornfels) were made. 477 Identifying the Decorative Stone Samples from the Mining Museum’s Collection Identifying the Decorative Stone Samples from the Mining Museum’s Collection Identifying the Decorative Stone Samples from the Mining Museum’s Collection 4
Conclusions Thus, preliminary studies allowed us to outline the characteristic fields of the studied
decorative rocks samples on their bulk composition plots. To obtain accurate results, it
is necessary to expand the number of samples under study and add reference samples to
the diagram. The lack of variation in the petrogenic elements content level in decorative
rocks of different types and geographic reference also requires the analysis of not only
chemical but also mineralogical composition of rocks. Driven by the need in non-
destructive techniques, it is planned to use Raman spectroscopy to make mineralogical
composition maps of the samples. 3
Results and Discussion The absence of reference samples let us consider the obtained results as preliminary. Nevertheless, on the ternary plots of the rock samples bulk composition, it is possible
to outline the characteristic fields corresponding to certain samples groups. The Ca-Fe-Si diagram illustrates obvious trends: visible outlining of marble,
Urazovskaya jasper, and andesite characteristic fields. The bimodal distribution of
points corresponding to green marble possibly occurred due to the presence of large
calcite phenocrysts, the content of which varies not only during the transition from
sample to sample, but also unevenly distributed over the analyzed area of the sample. On the Ca-Fe-K triangular plot, the characteristic field of Urazovskaya jasper shows
that it is ferruginous which is also expressed in its characteristic purple-red color
(Fig. 1). The field corresponding to the decorative andesites and the bimodality of the
green marble analysis distribution are presented on all charts and can later serve as a
diagnostic feature. Fig. 1. Ternary plots of the bulk chemical composition of polished decorative stones from the
collection of Empress Catherine the Great, plotted according to the data acquired by handheld
XRF analyzer Delta Olympus. Rock types and characteristic fields: 1 - Korogon porphyry; 2 -
Tigiretsky quartz; 3, I - Urazovskaya jasper; 4, II - green marble; 5, III - marble; 6 - andesite; 7 -
hornfels Fig. 1. Ternary plots of the bulk chemical composition of polished decorative stones from the
collection of Empress Catherine the Great, plotted according to the data acquired by handheld
XRF analyzer Delta Olympus. Rock types and characteristic fields: 1 - Korogon porphyry; 2 -
Tigiretsky quartz; 3, I - Urazovskaya jasper; 4, II - green marble; 5, III - marble; 6 - andesite; 7 -
hornfels 478 N. Borovkova and M. Machevariani Borovkova NV (2017) Personal mineralogical collection of Empress Catherine the great in the
mining museum collection. Bulletin of St. Petersburg State University of Technology and
Design, №1, Series 2. Arts Philology 8–15 © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 479–482, 2019.
https://doi.org/10.1007/978-3-030-22974-0_118 Reference Borovkova NV (2017) Personal mineralogical collection of Empress Catherine the great in the
mining museum collection. Bulletin of St. Petersburg State University of Technology and
Design, №1, Series 2. Arts Philology 8–15 Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 1
Introduction Preservation of the monuments of cultural heritage is one of the priorities of the modern
society. This problem becomes especially acute where the monuments are exhibited in
the open air and subjected to destructive effects of the environment. In large cities, such
as St. Petersburg, the deterioration of natural stone is notably fast, which is primarily
due to the influence of the anthropogenic factor (The Effect 2019). Now we present the
results of a multi-year, comprehensive study of the state of historical stone monuments
of St. Petersburg, which are exposed to the destructive impact of the urban environ-
ment. The obtained data are of exceptional scientific interest for studying the processes
of stone deterioration under the impact of the environment. Monitoring of the State of St. Petersburg Stone
Monuments and the Strategy
of Their Preservation O. Frank-Kamenetskaya1(&), D. Vlasov1, V. Rytikova2,
V. Parfenov3, V. Manurtdinova2, and M. Zelenskaya1 1 St. Petersburg State University, St. Petersburg, Russia
ofrank-kam@mail.ru
2 State Museum of Urban Sculpture, St. Petersburg, Russia 1 St. Petersburg State University, St. Petersburg, Russia
ofrank-kam@mail.ru
2 State Museum of Urban Sculpture, St. Petersburg, Russia
3 St. Petersburg Electrotechnical University «LETI», St. Petersburg, Russia 1 St. Petersburg State University, St. Petersburg, Russia
ofrank-kam@mail.ru
2 State Museum of Urban Sculpture, St. Petersburg, Russia
3 St. Petersburg Electrotechnical University «LETI», St. Petersburg, Russia 3 St. Petersburg Electrotechnical University «LETI», St. Petersburg, Russia Abstract. The results of the multi-year monitoring of the state of Saint
Petersburg stone monuments are summarized. The unique collection of deco-
rative stones in museum Necropolis and the deposits that were most likely used
to create them are studied. The processes of stone monuments’ degradation in
response to physical, chemical and biogenic influences are discussed. Special
attention is paid to describing the monitoring methodology and the structure of
the monitoring information database. Drawing on received results, the strategy
for the conservation and restoration of monuments are discussed. The obtained
data are of exceptional scientific interest in studying the processes of stone
deterioration under the impact of the environment. Keywords: Cultural heritage Monitoring Stone deterioration
Anthropogenic weathering Restoration and conservation works 2
Methods and Approaches Monitoring studies have been carried out in the Historical Necropoleis of the Museum
of Urban Sculpture since 1998, where on a small square there is a unique collection of
decorative and facing stone. The stone is intensively destroyed due to destructive O. Frank-Kamenetskaya et al. 480 influence of the volatile and humid Petersburg climate and unfavorable ecological
situation. In this work, in addition to the Saint Petersburg scientists, museum staff and
restorers, post-graduate students and students of the St. Petersburg State University, the
Russian and Herzen State Pedagogical University took part. Over the past years, more
than 1300 monuments of the Museum Necropolis have been examined (some of them
several times), Based on the results obtained, a methodology for monitoring studies of
stone materials of monuments was developed, which included the following steps: 1. Visual inspection of the object. Photographic documentation. Sampling. 2. Qualimetric
evaluation of the integral state of the monument material (performed for 348 monu-
ments). 3. Mapping of the types of material deterioration. 4. Examination of the
samples of material and products of its deterioration by instrumental procedures
(petrographic description of thin sections under a polarizing microscope, SE micro-
scopy with EDX, X-ray phase analysis, biological methods). 5. Examination of the
species composition of the microbial community on the surface of the monument. 6. Developing a 3D model of the monument and a quantitative estimate of the types of
destruction of its material by the results of laser scanning. 7. Study of the local cor-
rosivity of the air environment near the monuments. 8. Archival research. 9. Creating
and maintaining a database on the state of the sculptural monuments in St. Petersburg. 3
Results and Discussion Stone Material of Monuments. The diverse stone material in the museum Necropoleis
is represented by marbles, limestones, granites and other hard rocks (gneisses, gab-
broids, amphibolites, quartzites). The museum Necropoleis are not inferior to the
historical center of St. Petersburg in the variety of stone. Basically, the stone came from
Italy and the areas close to St. Petersburg (from the territory of the present Leningrad
region, Karelia and Finland). Qualimetric Evaluation of the Integral State of the Monument Material. The technique
was developed jointly with V.M. Marugin (VITU, SPb). It was shown that the degree of
stone destruction in the museum Necropoleis varies from 2 to 51%. In most cases, the
extent of carbonate rock deterioration does not exceed 25% and that of granite and other
hard silicate rocks - 10%. This is due to the considerable contribution of chemical
weathering (formation of gypsum-enriched patina) in the deterioration of memorials of
marble and limestone. Cracks occur on the surface of carbonate rocks that are hetero-
geneous in composition and structure (Ruskeala, Italian breccia and brecciated marbles,
Pudost and Putilovo limestones) at least 10% more often than on other denser and more
homogeneous marbles and limestones. But on denser solid silicate rocks (granites, etc.),
cracks occur no less frequently than on carbonate rocks. At the same time, they are much
more common (found on 80% of monuments) on such dense homogeneous rocks as
Serdobol granite and Shokshinsky quartzite, which indicates a possibility of their
anthropogenic or constructional origin. The incidence of the primary gypsum crust on
the surface of limestones is more frequent than on the surface of marbles. Among
limestones, the gypsum-rich patina is most often found on the surface of the porous
Pudost travertine (on the surfaces of 50% of examined monuments). Among marbles, it Monitoring of the State of St. Petersburg Stone Monuments 481 is most often seen on the homogeneous Carrara marble (on the surfaces of 26% of the
surveyed monuments). Its detachment together with marble and the formation of a
secondary gypsum crust are observed only on the monuments with a complex surface
relief made of dense homogeneous marble: (white Carrara and light gray Bardiglio). In
fouling, biofilms with dominant fungi are widespread on the surface of all rocks The
input of microorganisms (fungi, algae, lichens) in rock deterioration varies from 2 to
10%. 4
Conclusion Integrated monitoring the state of St. Petersburg monuments provided an objective
picture of the state of the their materials, makes it possible to take timely interventions for
the restoration and conservation of works of art, to plan the necessary measures to protect
the stone from deterioration and in result make it possible to preserve and adequately
exhibit the works of monumental sculpture and memorial art of St. Petersburg, which are
an impressive, imaginative part of the world history and culture. Acknowledgements. This study was supported by RSF project no 19-17-00141 and performed
using the equipment of the SPBU resource centers “X-Ray Diffraction Methods for Studying
Matter,” “Nanotechnologies,” and “Geomodel”. 3
Results and Discussion The degree of manifestation of various types of stone destruction significantly
varies depending on the exhibiting conditions of the monument, the characteristics of
the stone material, as well as the timing and effectiveness of work on the care. Mapping of Deterioration Forms. Ultrasonic sounding was used to detect hetero-
geneities of the rock material invisible from the surface. Method for monitoring the
biofouling of cultural heritage sites using computer technology allowing to register the
areas of the most threatened biodeterioration sites was developed (Fig. 1). Beside 3D
laser scanning method was used for to create 3D computer models and to carry out the
quantitative measurements of various kinds of damage of the monument materials:
cracks, chips, scratches, gypsum crusts (Fig. 2) and others. Database on the State of the Sculptural Monuments of St. Petersburg. One of the most
important stages in monitoring the state of the monuments was creating and populating
a specialized database used to store, analyze and structure the accumulated factual
information Currently, the database includes characteristics of the state of 650 stone
monuments in the Necropoleis of the Museum of Urban Sculpture and in other parts of
St. Petersburg. Approaches and Methods of Monument Protection from Damage. To assess the
effectiveness and safety of different approaches when removing biofilms, mud buildups
and gypsum crusts from the surface of stone monuments, a comparative analysis was
made of the potential of various chemical biocidal treatments and of laser cleaning
options. The results of the experiments showed that the laser cleaning technology for
removal of biofilms from the surface of the stone is comparable, and in some cases
even superior to chemical treatment with hydrogen peroxide and kaolin. In the case of
intensive development of biofouling, containing mosses and lichens, the efficiency of
laser cleaning is significantly higher than the efficiency of chemical biocidal treatment. The use of laser cleaning to remove gypsum-rich patina is also effective. a
b
Fig. 1. Biofilms with dominant algae (color yellow) and dark-colored micromycetes (color
brown): a-photograph, b-map b Fig. 1. Biofilms with dominant algae (color yellow) and dark-colored micromycetes (color
brown): a-photograph, b-map 482 O. Frank-Kamenetskaya et al. Fig. 2. Electronic 3D-model of fragment of the mourner sculpture, on which the area of the
gypsum crust is highlighted Fig. 2. Electronic 3D-model of fragment of the mourner sculpture, on which the area of the
gypsum crust is highlighted Ceramics Sugar Jars Pieces
from Aveiro Production S. Moutinho1(&), C. Costa1,2, Â. Cerqueira2, C. Sequeira2,
D. Terroso2, J. Nobre1, P. Morgado2, A. Velosa1,2, and F. Rocha1,2 1 RISCO, Civil Engineering Department, University of Aveiro,
3810-193 Aveiro, Portugal
sara.moutinho@ua.pt
2 GeoBioTec, Geosciences Department, University of Aveiro,
3810-193 Aveiro, Portugal Abstract. Ceramics sugar jars pieces are, from morphological point of view,
conical containers of fired clay with a hole in the vertex, which were used to
sugar cane pulp maturation into sugar cake. These ceramic materials were
produced in Aveiro given the existence of local raw material. Also, this
occurrence of geological deposits exploited for red clays allowed the local
development of strong pottery production center, transforming the city of Aveiro
into one of the major Portuguese cultural heritage sites very rich in traditional
ceramic tiles (azulejos) and other ceramic products. After local manufacture, the
ceramic sugar jars pieces were exported as sugar production devices for Madeira
island, Cape Verde archipelago and later, for Brazil. Also, these materials were
found in buildings construction. So, this work focuses on the characterization of
ceramic sugar jars produced in Aveiro and its construction use comparing with
properties of other ceramics, justifying their preference for export to several
countries of the world. Keywords: Ceramic products Sugar jars pieces Properties
Construction elements Keywords: Ceramic products Sugar jars pieces Properties
Construction elements Reference Frank-Kamenetskaya OV, Vlasov DY, Rytikova VV (Ed) (2019) The Effect of the Environment
on Saint Petersburg’s Cultural Heritage. Springer, Switzerland Frank-Kamenetskaya OV, Vlasov DY, Rytikova VV (Ed) (2019) The Effect of the Environment
on Saint Petersburg’s Cultural Heritage. Springer, Switzerland Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. 2
Methods and Approaches Mineralogical analysis was carried out by X-Ray diffraction, using a Panalytical
X’Pert-Pro MPD, Ka Cu (k = 1,5405 Å) radiation on random-oriented powders;
chemical composition was assessed by X-Ray Fluorescence using a Panalytical Axios
PW4400/40 X-Ray Fluorescence spectrometer for major and trace elements and Lost on
Ignition (LOI) was also determined. Compressive strength was assessed by a Shimadzu:
AG-IC equipment. TGA analysis was also performed. 1
Introduction The use of ceramic sugar jars pieces in ancient masonry walls in the Aveiro district
reflects the use of these materials in construction beyond the production and trans-
portation of sugar (Nobre 2017). Ceramic sugar jars materials were produced in Aveiro
given the existence of raw material in abundance and of very good quality. In the
fiftieth century the production center of Barreiro (close to Lisbon) would have ceased
production and until the independence of Brazil, in the early nineteenth century, Aveiro
would have been the only producer of these ceramic materials in Portugal (Morgado
2014). This production will have provided intense trade with the major sugar producing
centers. Due to the local absence of natural stone for construction, the rejected/surplus
ceramics were used as building material on the walls. Recently, following old house
demolitions in the city of Aveiro, whole walls have been discovered with these ceramic © The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 483–484, 2019. https://doi.org/10.1007/978-3-030-22974-0_119 S. Moutinho et al. 484 materials, many of which were practically intact, which allowed the development of
this comparative study. 3
Results and Conclusions The chemical and mineralogical properties of ceramics were similar, pointing to local
production using only local raw materials. Quartz is present in all samples. The
phyllosilicates are not present in any sample of the sugar ceramics but are present in all
the remaining ceramic samples. The presence/absence of phyllosilicates is an indicator
of the heating process temperature, higher on the case of the sugar ceramic jars. The
compressive strength analysis of the ceramics sugar jars pieces shows higher values
(mean 9.5 MPa) than other ceramics (mean 8.0 MPa). References Morgado P, Rocha F (2014) Produção da cerâmica do açúcar em Aveiro pode explicar origem
dos Ovos moles. Univ Aveiro J Nobre J, Faria P, Velosa AL (2017) Paredes pão-de-açúcar em edifícios de Aveiro Evolu
materiais e características. Master thesis, New University of Lisbon Nobre J, Faria P, Velosa AL (2017) Paredes pão-de-açúcar em edifícios de Aveiro Evolução,
materiais e características. Master thesis, New University of Lisbon Nobre J, Faria P, Velosa AL (2017) Paredes pão-de-açúcar em edifícios de Aveiro Evolução,
materiais e características. Master thesis, New University of Lisbon Open Access This chapter is licensed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing,
adaptation, distribution and reproduction in any medium or format, as long as you give appro-
priate credit to the original author(s) and the source, provide a link to the Creative Commons
license and indicate if changes were made. The images or other third party material in this chapter are included in the chapter’s Creative
Commons license, unless indicated otherwise in a credit line to the material. If material is not
included in the chapter’s Creative Commons license and your intended use is not permitted by
statutory regulation or exceeds the permitted use, you will need to obtain permission directly
from the copyright holder. Author Index A
Abarzúa, G., 3
Abou Zahr Diaz, M., 443
Afanasiev, V., 179
Agapov, I., 15, 19, 24, 29
Akhmetzyanova, M., 323
Akpınar, İ., 7
Alashanov, A., 385
Alaskhanov, A., 360, 365, 369
Alawiyeh, M. A., 443
Aleksandrova, T., 59
Aliev, S., 335
Anchugova, E., 281
Andreicheva, L., 11
Andrejkovičová, S., 381
Anisimov, I., 15, 19, 24, 29
Arduin, D., 313
Arsent’ev, K., 423
Arsentyev, V., 66
Artamonov, A., 323
Askhabov, A., 33
Astakhova, Yu., 119
Aupova, N., 107
B
Babaev, V., 389
Balykov, A., 307, 372
Bedina, V., 277
Belyanin, D., 195
Berkh, K., 37
Bilskaya, I., 140
Bolatov, A., 267
Bondarenko, D., 286
Borovkova, N., 475
Brodskaya, R., 140
Bulaev, A., 189
Burtsev, I., 41, 192
C
Cerqueira, Â., 311, 313, 381, 483
Chanturiya, V., 45
Chekushina, T., 457, 461
Chernysheva, N., 315
Chikisheva, T., 49
Chulkova, I., 319
Çiftçi, E., 7, 53, 84, 183
Costa, C., 311, 313, 381, 483
D
Dai, Q., 413, 463
Danyushevsky, L., 107
Demyanova, V., 348
Deng, S., 463
Denisova, Y., 235
Deryabin, P., 319
Diarra, K., 53
Dobrinskaya, O., 272
Dolotova, A., 15, 24, 29
Dong, F., 290, 413, 463
Doroganov, V., 296
Drobe, M., 37
E
Elbendari, A., 59
Eliseev, A., 179
Elistratkin, M., 315
© The Editor(s) (if applicable) and The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 485–488, 2019. https://doi.org/10.1007/978-3-030-22974-0 A
Abarzúa, G., 3
Abou Zahr Diaz, M., 443
Afanasiev, V., 179
Agapov, I., 15, 19, 24, 29
Akhmetzyanova, M., 323
Akpınar, İ., 7
Alashanov, A., 385
Alaskhanov, A., 360, 365, 369
Alawiyeh, M. A., 443
Aleksandrova, T., 59
Aliev, S., 335
Anchugova, E., 281
Andreicheva, L., 11
Andrejkovičová, S., 381
Anisimov, I., 15, 19, 24, 29
Arduin, D., 313
Arsent’ev, K., 423
Arsentyev, V., 66
Artamonov, A., 323
Askhabov, A., 33
Astakhova, Yu., 119
Aupova, N., 107
B
Babaev, V., 389
Balykov, A., 307, 372
Bedina, V., 277
Belyanin, D., 195
Berkh, K., 37
Bilskaya, I., 140
Bolatov, A., 267
Bondarenko, D., 286
© The Editor(s) (if applicable) an
S. © The Editor(s) (if applicable) and The Author(s) 2019
S. Glagolev (Ed.): ICAM 2019, SPEES, pp. 485–488, 2019.
https://doi.org/10.1007/978-3-030-22974-0 G Garkavi, M., 323 Gavshina, O., 296 Gerasimov, A., 66, 70 Klimenko, T., 94 Golubeva, I., 192 Gomes, A., 231 Klimenko, V., 94 Klimova, L., 300 Gomze, L. A., 133 Klyucharev, D., 453 Goncharov, A., 327 Kobayashi, Y., 451 Kolesov, E., 49 Kolodezhnaya, E., 323
Koneev, R., 99 Kononov, O., 152 Kononova, N., 267 Konovalov, V., 327 Gzogyan, T., 75, 80 Korneev, S., 198 H
Hadisov, V., 369
Hajjaji, W., 381
Hardaev, P., 315
Harja, M., 255
Huang, J., 290
Huang, Y., 290
Hubaev, M., 335
Hui, A., 419
Huo, T., 290
I
Idir, R., 339
Ignatyev, G., 281
Ilyina, V., 259
Iospa, A., 119
Iskandarov, N., 99
Ismailova, Z., 385
Izatulina, A., 415
J
Javid, F., 84 H Korovkin, D., 307, 372 Kotova, E., 70 Kotova, I., 127 Kotova, O., 103, 133, 255, 281 Koulibaly, M., 143 Kovalev, S., 356 Kovalevski, V., 201 Kovalyov, S., 352 Kozhukhova, N., 331 Krivosheeva, A., 99 Krivovichev, S., 447 Kuznetsov, A., 267 L
Labuzova, M., 376
Large, R., 107
Larionov, P. M., 432
Lashina, I., 344
Lawrence, K., 445 Author Index Glagolev (Ed.): ICAM 2019, S
https://doi.org/10.1007/978-3-030 Borovkova, N., 475
Brodskaya, R., 140
Bulaev, A., 189
Burtsev, I., 41, 192 Borovkova, N., 475
Brodskaya, R., 140
Bulaev, A., 189
Burtsev, I., 41, 192 C
Cerqueira, Â., 311, 313, 381, 483
Chanturiya, V., 45
Chekushina, T., 457, 461
Chernysheva, N., 315
Chikisheva, T., 49
Chulkova, I., 319
Çiftçi, E., 7, 53, 84, 183
Costa, C., 311, 313, 381, 483 Alawiyeh, M. A., 443 Aleksandrova, T., 59 Andrejkovičová, S., 381 D E
Elbendari, A., 59
Eliseev, A., 179
Elistratkin, M., 315 Author Index 486 Elkina, Yu., 189
Evtushenko, E., 277, 292, 296 F
Fedorov, A., 327
Fischer, H.-B., 344
Fomenko, I., 466
Frank-Kamenetskaya, O., 415, 479 G
Galdina, V., 319
Garkavi, M., 323
Gavshina, O., 296
Gerasimov, A., 66, 70
Ghaboura, M., 443
Glagolev, E., 315, 344, 395
Golubeva, I., 192
Gomes, A., 231
Gomze, L. A., 133
Goncharov, A., 327
Gorbatova, E., 167
Gorobtsov, D., 466
Grakova, O., 235
Gromilov, S., 179
Gubareva, E., 376
Gurova, E., 319
Gusev, V., 195
Gutiérrez, L., 3
Gzogyan, S., 75, 80
Gzogyan, T., 75, 80 L J
Javid, F., 84 J
Javid, F., 84 487 Author Index Lazareva, E., 208
Lazareva, V., 66
Le Saout, G., 339
Lesovik, V., 315, 344, 395
Levchenko, E., 453
Li, Q., 413, 463
Likhnikevich, E., 119
Lipko, I., 423
Lipko, S., 423
Loganina, V., 348
Logovskaya, G., 198
Loktionov, V., 292
Luo, Y., 413, 463
Lyahnitskaya, V., 140 Lazareva, E., 208
Lazareva, V., 66
Le Saout, G., 339
Lesovik, V., 315, 344, 395
Levchenko, E., 453
Li, Q., 413, 463
Likhnikevich, E., 119
Lipko, I., 423
Lipko, S., 423
Loganina, V., 348
Logovskaya, G., 198
Loktionov, V., 292
Luo, Y., 413, 463
Lyahnitskaya, V., 140 Nobre, J., 483
Novikova, S., 172 O
Ogurtsova, Y., 376
Ogurtsova, Yu., 403
Olmaskhanov, N., 219
Onosov, D., 248
Otake, T., 445, 451
Ozhogina, E., 103, 119, 167
P
Pakhomova, A., 447
Parfenov, V., 479
Pateyuk, S., 90
Pavlenko, A., 123
Pavlenko, V., 94
Pavlov, V., 136
Perovskiy, I., 41
Piryaev, A., 208
Podolian, E., 127
Pokhilenko, N., 179
Ponaryadov, A., 192, 255
Ponomarchuk, V., 208
Potemkin, V., 59
Prokopyev, E., 49
Prokopyev, S., 49
Proshlyakov, A., 457
Pursheva, A., 323
Pystin, A., 129
Pystina, Y., 129
R
Rammlmair, D., 37, 244
Rashchupkina, M., 319
Razmyslov, I., 133
Rocha, F., 311, 313, 381, 483
Rogozhin, A., 119, 167
Roux, J.-C., 339
Ryabova, A., 300
Rykunova, M., 403
Rytikova, V., 479
S
Sagitova, A., 15, 19, 24, 29
Saidumov, M., 369
Salamanova, M., 360, 385
Sangu, E., 53
Sato, T., 445, 451
Saydumov, M., 360, 365
Sazonov, A., 136
Selim, H., 183
Sendir, H., 183 O P
Pakhomova, A., 447
Parfenov, V., 479
Pateyuk, S., 90
Pavlenko, A., 123
Pavlenko, V., 94
Pavlov, V., 136
Perovskiy, I., 41
Piryaev, A., 208
Podolian, E., 127
Pokhilenko, N., 179
Ponaryadov, A., 192, 255
Ponomarchuk, V., 208
Potemkin, V., 59
Prokopyev, E., 49
Prokopyev, S., 49
Proshlyakov, A., 457
Pursheva, A., 323
Pystin, A., 129
Pystina, Y., 129 M M
Ma, B., 290
Machevariani, M., 475
Malyukov, V., 221
Manurtdinova, V., 479
Markarova, M., 281
Masanin, O., 389
Maslennikov, V., 107
Maslennikova, S., 107
Matveeva, T., 45
Matyukhin, P., 239
Mazhitov, E., 348
Meima, J., 37
Melamud, V., 189
Melnikova, E., 189
Milman, B., 15
Min’ko, N., 272
Minibaev, A., 205
Mirsamiev, N., 219
Mishin, D., 352, 356
Morales, J., 3
Moreva, I., 277, 292
Morgado, P., 483
Moutinho, S., 311, 313, 3
Mu, B., 427
Mugisho, J., 219
Murtazaev, S.-A., 335, 360
Murtazaeva, T., 369
N M
Ma, B., 290
Machevariani, M., 475
Malyukov, V., 221
Manurtdinova, V., 479
Markarova, M., 281
Masanin, O., 389
Maslennikov, V., 107
Maslennikova, S., 107
Matveeva, T., 45
Matyukhin, P., 239
Mazhitov, E., 348
Meima, J., 37
Melamud, V., 189
Melnikova, E., 189
Milman, B., 15
Min’ko, N., 272
Minibaev, A., 205
Mirsamiev, N., 219
Mishin, D., 352, 356
Morales, J., 3
Moreva, I., 277, 292
Morgado, P., 483
Moutinho, S., 311, 313, 381, 483
Mu, B., 427
Mugisho, J., 219
Murtazaev, S.-A., 335, 360, 365, 385
Murtazaeva, T., 369
N
N kh
M
365 R
Rammlmair, D., 37, 244
Rashchupkina, M., 319
Razmyslov, I., 133
Rocha, F., 311, 313, 381, 483
Rogozhin, A., 119, 167
Roux, J.-C., 339
Ryabova, A., 300
Rykunova, M., 403
Rytikova, V., 479 Minibaev, A., 205 Mirsamiev, N., 219 Mishin, D., 352, 356 Moutinho, S., 311, 313, 381, 483 S
Sagitova, A., 15, 19, 24, 29
Saidumov, M., 369
Salamanova, M., 360, 385
Sangu, E., 53
Sato, T., 445, 451
Saydumov, M., 360, 365
Sazonov, A., 136
Selim, H., 183
Sendir, H., 183 S
Sagitova, A., 15, 19, 24, 29
Saidumov, M., 369
Salamanova, M., 360, 385
Sangu, E., 53
Sato, T., 445, 451
Saydumov, M., 360, 365
Sazonov, A., 136
Selim, H., 183
Sendir, H., 183 N Z., 471
Zhao, X., 463
Zhao, Y., 290
Zhernovsky, I., 331, 389, 403, 407
Zhmodik, S., 195, 208
Zhu, Y., 436
Zhukova, V., 119
Zvyagina, E., 136
488
Auth Uralbekov, B., 267
Usikov, S., 389
Ustinov, I., 70, 152 V
Vaisberg, L., 146, 152
Vdovina, I., 157
Velosa, A., 313, 483
Vikhot, A., 235
Vlasov, D., 479
Volodchenko, A., 344, 399
Volodin, V., 307, 372
Vorobev, A., 227
Vorobyev, K., 221, 231
Vorobyov, N., 214
Voytekhovsky, Yu., 162 Spiridonov, I., 453 Stefanovsky, S., 447 Sun, H. Y., 471
Sun, S., 255, 290
Svetlichnyi, V., 267
Sycheva, N., 119
Sysa, E., 296
Sysa, O., 277, 292 Y
Yakushina, O., 119, 167
Yarg, L., 466
Yashkina, S., 296
Yastrebinskiy, R., 123
Yatimov, U., 107
Yatsenko, E., 300
Yu, H., 290
Yudin, D., 172
Yudintsev, S., 447
Yusupov, T., 172 T
Tanoh Boguy, E. M., 461
Tauson, V., 423
Tcharo, H., 143
Tchibozo, F. K. N., 143
Terroso, D., 311, 313, 483
Timonina, N., 223
Titov, A., 432
Titov, S., 393
Toda, K., 451
Tolstoy, A., 395
Travin, A., 172
Troshina, O., 19
Tseluyko, A., 107
Tukuser, V., 49
Tursunkulov, O., 99
U
Ugapieva, S., 179
Uljasheva, N., 235 T Tcharo, H., 143 Terroso, D., 311, 313, 483 N Nakhaev, M., 365
Nelyubova, V., 389, 403
Nesterenko, G., 195
Nikiforova, Z., 111, 115
Nikitina, L., 90
Nikolaeva, N., 59
Nikonow, W., 244
Nizina, T., 307, 372 Nakhaev, M., 365
Nelyubova, V., 389, 403
Nesterenko, G., 195
Nikiforova, Z., 111, 115
Nikitina, L., 90
Nikolaeva, N., 59
Nikonow, W., 244
Nizina, T., 307, 372 488 Author Index Senyut, V., 179
Sequeira, C., 311, 313, 483
Shadrunova, I., 457
Shchemelinina, T., 255, 281
Shchesnyak, E., 227
Shchiptsov, V., 201
Shelukhina, I., 127
Shevchenko, S., 140
Shevchenko, V., 267
Shmakova, A., 192, 214
Shushkov, D., 281
Sigida, A., 99
Silaev, V., 133
Silyanov, S., 136
Smetannikov, A., 248
Sobolev, K., 331
Sokolov, S., 212
Spiridonov, I., 453
Stefanovsky, S., 447
Strokova, V., 286, 331, 376, 389, 399, 403,
407
Sun, H. Y., 471
Sun, S., 255, 290
Svetlichnyi, V., 267
Sycheva, N., 119
Sysa, E., 296
Sysa, O., 277, 292
T
Tanoh Boguy, E. M., 461
Tauson, V., 423
Tcharo, H., 143
Tchibozo, F. K. N., 143
Terroso, D., 311, 313, 483
Timonina, N., 223
Titov, A., 432
Titov, S., 393
Toda, K., 451
Tolstoy, A., 395
Travin, A., 172
Troshina, O., 19
Tseluyko, A., 107
Tukuser, V., 49
Tursunkulov, O., 99
U
Ugapieva, S., 179
Uljasheva, N., 235
Uralbekov, B., 267
Usikov, S., 389
Ustinov, I., 70, 152
V
Vaisberg, L., 146, 152
Vdovina, I., 157
Velosa, A., 313, 483
Vikhot, A., 235
Vlasov, D., 479
Volodchenko, A., 344, 399
Volodin, V., 307, 372
Vorobev, A., 227
Vorobyev, K., 221, 231
Vorobyov, N., 214
Voytekhovsky, Yu., 162
W
Wang, A., 419, 427, 436
Wang, F., 413, 436, 463
Wang, K., 290
Wang, W., 436
Y
Yakushina, O., 119, 167
Yarg, L., 466
Yashkina, S., 296
Yastrebinskiy, R., 123
Yatimov, U., 107
Yatsenko, E., 300
Yu, H., 290
Yudin, D., 172
Yudintsev, S., 447
Yusupov, T., 172
Z
Zagorodniuk, L., 395
Zaikovskii, V., 432
Zelenskaya, M., 415, 479
Zhang, A., 427
Zhang, L. Z Z
Zagorodniuk, L., 395
Zaikovskii, V., 432
Zelenskaya, M., 415, 479
Zhang, A., 427
Zhang, L. Z., 471
Zhao, X., 463
Zhao, Y., 290
Zhernovsky, I., 331, 389, 403, 407
Zhmodik, S., 195, 208
Zhu, Y., 436
Zhukova, V., 119
Zvyagina, E., 136 Zagorodniuk, L., 395
Zaikovskii, V., 432
Zelenskaya, M., 415, 479
Zhang, A., 427
Zhang, L. Z., 471
Zhao, X., 463
Zhao, Y., 290
Zhernovsky, I., 331, 389, 403, 407
Zhmodik, S., 195, 208
Zhu, Y., 436
Zhukova, V., 119
Zvyagina, E., 136
|
https://openalex.org/W4361902241
|
https://figshare.com/articles/journal_contribution/Supplementary_Figure_2B_from_Genetic_Variations_in_Multiple_Drug_Action_Pathways_and_Survival_in_Advanced_Stage_Non_Small_Cell_Lung_Cancer_Treated_with_Chemotherapy/22442424/1/files/39893316.pdf
|
unk
| null |
Supplementary Figure 1B from Genetic Variations in Multiple Drug Action Pathways and Survival in Advanced Stage Non–Small Cell Lung Cancer Treated with Chemotherapy
| null | 2,023
|
cc-by
| 12
|
p-value=0
Time (years)
2
3
4 p-valu
Time (years)
2
3
4
|
https://openalex.org/W3125631368
|
https://www.itm-conferences.org/articles/itmconf/pdf/2021/01/itmconf_icmsa2021_03004.pdf
|
Latin
| null |
Total edge irregularity strength of quadruplet and quintuplet book graphs
|
ITM web of conferences
| 2,021
|
cc-by
| 7,862
|
1 Introduction Graph labelling is a function of the set of integers to the set of elements on the graph (vertices,
edges or both) with certain conditions [1]. Irregular edge k-labelling was introduced by
Chartrand et al. [2] as a function of the set of edges to the set {1,2, … , 𝑘} such that any two
different vertices in graph 𝐺 have different weights. Let 𝑣 be a vertex ; the weight of 𝑣 is the
sum of labels of edges that are incident to vertex 𝑣 . If graph 𝐺 can be labelled with an
irregular edge 𝑘-labelling, then the minimum 𝑘 is called irregularity strength of 𝐺(denoted
by 𝑠(𝐺)). Bača et al. defined an edge irregular total 𝑘-labelling of graph 𝐺 as a function 𝑓 from the
union of the set of vertices and the set of edges to the set {1,2, … , 𝑘} such that any two
different edges of 𝐺 have different weights [3]. Let 𝑥𝑦 be an edge. The weight of the edge
𝑥𝑦 (denoted by 𝜔𝑓(𝑥𝑦)) is 𝜔𝑓(𝑥𝑦) = 𝑓(𝑥) + 𝑓(𝑦) + 𝑓(𝑥𝑦). If the graph 𝐺 can be labelled
with a total irregular 𝑘-labelling, then the minimum 𝑘 is called the total edge irregularity
strength 𝐺 (denoted by 𝑡𝑒𝑠(𝐺)). In [3], Bača et al. have also given the lower bound of 𝑡𝑒𝑠(𝐺). I
d J
d
l i
[4] h
d t
i
d 𝑡
f t
R
h
th 𝑡
f
li with a total irregular 𝑘-labelling, then the minimum 𝑘 is called the total edge irregularity
strength 𝐺 (denoted by 𝑡𝑒𝑠(𝐺)). In [3], Bača et al. have also given the lower bound of 𝑡𝑒𝑠(𝐺). Ivanco and Jendrol in [4] have determined 𝑡𝑒𝑠 of trees. Research on the 𝑡𝑒𝑠 of cyclic
graphs for various graph classes is still being done. Several studies on the exact value of 𝑡𝑒𝑠
in some cyclic graphs, including some book graphs, have been conducted by [5-19]. Ivanco and Jendrol in [4] have determined 𝑡𝑒𝑠 of trees. Research on the 𝑡𝑒𝑠 of cyclic
graphs for various graph classes is still being done. Several studies on the exact value of 𝑡𝑒𝑠
in some cyclic graphs, including some book graphs, have been conducted by [5-19]. *Corresponding author: ratnasari.lucia@gmail.com ITM Web of Conferences 36, 03004 (2021)
ICMSA 2020 ITM Web of Conferences 36, 03004 (2021)
ICMSA 2020 https://doi.org/10.1051/itmconf/20213603004 Total edge irregularity strength of quadruplet
and quintuplet book graphs
Lucia Ratnasari1*, Sri Wahyuni1, Yeni Susanti1, and Diah Junia Eksi Palupi1
1Department of Mathematics, Universitas Gadjah Mada, Yogyakarta, Indonesia Lucia Ratnasari1*, Sri Wahyuni1, Yeni Susanti1, and Diah Junia Eksi Palupi1
1Department of Mathematics, Universitas Gadjah Mada, Yogyakarta, Indonesia Abstract. Let G= (V, E) be a finite, simple and undirected graph with a
vertex set V and an edge set E. An edge irregular total k-labelling is a
function f : V E → {1,2,…,k} such that for any two different edges xy and
x'y' in E, their weights are distinct. The weight of edge xy is the sum of label
of edge xy, labels of vertex x and of vertex y. The minimum k for which the
graph G admits an edge irregular total k-labelling is called the total edge
irregularity strength of G, denoted by tes(G). We have determined the total
edge irregularity strength of book graphs, double book graphs and triple
book graphs. In this paper, we show the exact value of the total edge
irregularity strength of quadruplet book graphs and quintuplet book graphs. © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons
Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). 1 Introduction In previous research [20], we have shown the 𝑡𝑒𝑠 of triple book graphs, and we have
constructed the formula of an edge irregular total k-labelling of the first book, the second
book and the third book. The results, the first book, the second book and the third book have ITM Web of Conferences 36, 03004 (2021)
ICMSA 2020 https://doi.org/10.1051/itmconf/20213603004 different edge labelling formulas. In this research, we investigate the formula of an edge
irregular total k-labelling and determine the 𝑡𝑒𝑠 of quadruplet and quintuplet book graphs. different edge labelling formulas. In this research, we investigate the formula of an edge
irregular total k-labelling and determine the 𝑡𝑒𝑠 of quadruplet and quintuplet book graphs. 2 Main result The vertex set of 4B (C ) is 𝑉(4𝐵𝑛(𝐶𝑚)) = {𝑢1, 𝑧1, 𝑧2, 𝑧3, 𝑣4} ∪{𝑥𝑖,𝑗
1 , 𝑥𝑖,𝑗
2 , 𝑥𝑖,𝑗
3 , 𝑥𝑖,𝑗
4 ∶𝑖= 1, … 𝑛; 𝑗= 1, … , 𝑚−2}
and the edge set of 4𝐵𝑛(𝐶𝑚) is 𝑉(4𝐵𝑛(𝐶𝑚)) = {𝑢1, 𝑧1, 𝑧2, 𝑧3, 𝑣4} ∪{𝑥𝑖,𝑗
1 , 𝑥𝑖,𝑗
2 , 𝑥𝑖,𝑗
3 , 𝑥𝑖,𝑗
4 ∶𝑖= 1, … 𝑛; 𝑗= 1, … , 𝑚−2}
and the edge set of 4𝐵𝑛(𝐶𝑚) is
𝐸(4𝐵𝑛(𝐶𝑚)) = {𝑢1𝑧1, 𝑧1𝑧2, 𝑧2𝑧3, 𝑧3𝑣4} ∪
{⋃
⋃
𝑢𝑥1
𝑚−2
𝑛
𝑥1 𝑥1
𝑥1
𝑧1
𝑧3𝑥4 𝑥4 𝑥4
𝑥4
𝑣4 } 𝐸(4𝐵𝑛(𝐶𝑚)) = {𝑢1𝑧1, 𝑧1𝑧2, 𝑧2𝑧3, 𝑧3𝑣4} ∪ 𝐸(4𝐵𝑛(𝐶𝑚)) = {𝑢1𝑧1, 𝑧1𝑧2, 𝑧2𝑧3, 𝑧3𝑣4} ∪ {⋃
⋃
𝑢𝑥𝑖,1
1
𝑚−2
𝑗=1
𝑛
𝑖=1
, 𝑥𝑖,𝑗
1 𝑥𝑖,𝑗+1
1
, 𝑥𝑖,𝑚−2
1
𝑧1, … , 𝑧3𝑥𝑖,1
4 𝑥𝑗,1
4 𝑥𝑖,𝑗+1
4
, 𝑥𝑖,𝑚−2
4
𝑣4 }. Similarly, we define quintuplet book graphs as follows: Similarly, we define quintuplet book graphs as follows: Definition 2.3. Let 𝐵𝑛
𝑞(𝐶𝑚), 1 ≤𝑞≤5 be the 𝑞𝑡ℎ copy of book graph Bn (Cm) as defined in
Definition 2.1. Let the vertices of 𝐵𝑛
𝑞(𝐶𝑚) be Definition 2.3. Let 𝐵𝑛
𝑞(𝐶𝑚), 1 ≤𝑞≤5 be the 𝑞𝑡ℎ copy of book graph Bn (Cm) as defined in
Definition 2.1. Let the vertices of 𝐵𝑛
𝑞(𝐶𝑚) be 𝑉(𝐵𝑛
𝑞(𝐶𝑚)) = {𝑢𝑞, 𝑣𝑞} ∪ {𝑥𝑖,𝑗
𝑞∶𝑖= 1, … , 𝑛; 𝑗= 1, … , 𝑚−2 }. A quintuplet book graph 5𝐵𝑛(𝐶𝑚) is a graph obtained from five copies of book graph
𝐵𝑛
𝑞(𝐶𝑚) by identifying vertex 𝑣𝑞 from book graph 𝐵𝑛
𝑞(𝐶𝑚) as vertex 𝑢𝑞+1 from book graph
𝐵𝑛
𝑞+1(𝐶𝑚) and renaming this vertex as 𝑧𝑞 , 1 ≤𝑞≤4. The vertex set of 5Bn(Cm) is The vertex set of 5Bn(Cm) is
𝑉(5𝐵𝑛(𝐶𝑚)) = {𝑢1, 𝑧1, 𝑧2, 𝑧3, 𝑧4, 𝑣5}
∪ {𝑥𝑖,𝑗
1 , 𝑥𝑖,𝑗
2 , 𝑥𝑖,𝑗
3 , 𝑥𝑖,𝑗,
4 𝑥𝑖,𝑗
5 ∶𝑖= 1, … , 𝑛; 𝑗= 1, … , 𝑚−2 }
and the edge set of 5𝐵𝑛(𝐶𝑚) is
𝐸(5𝐵𝑛(𝐶𝑚)) = {𝑢1𝑧1, 𝑧1𝑧2, 𝑧2𝑧3, 𝑧3𝑧4, 𝑧4𝑣5} ∪
{⋃
⋃
𝑢𝑥𝑖,1
1
𝑚−2
𝑗=1
𝑛
𝑖=1
, 𝑥𝑖,𝑗
1 𝑥𝑖,𝑗+1
1
, 𝑥𝑖,𝑚−2
1
𝑧1, … , 𝑧4𝑥𝑖,1
5 𝑥𝑗,1
5 𝑥𝑖,𝑗+1
5
, 𝑥𝑖,𝑚−2
5
𝑣5 }. We show the 𝑡𝑒𝑠 of quadruplet and of quintuplet book graphs in section 2.1. and section 2.2,
respectively. We show the 𝑡𝑒𝑠 of quadruplet and of quintuplet book graphs in section 2.1. 2 Main result We define the book graph, quadruplet book graph and quintuplet book graph as below: Definition 2.1. Let 𝐶𝑚
𝑖, 𝑖= 1, 2, … , 𝑛 be cycle graphs and the vertices of 𝐶𝑚
𝑖 be
𝑉(𝐶𝑚
𝑖) = {𝑢, 𝑣} ∪{𝑥𝑖,𝑗∶𝑗= 1, … , 𝑚−2} and the edges of 𝐶𝑚
𝑖 be
𝐸(𝐶𝑚
𝑖) = {𝑢𝑣, 𝑢𝑥𝑖1 , 𝑥𝑖,𝑗𝑥𝑖,𝑗+1, 𝑥𝑚−2𝑣∶𝑗= 1, … , 𝑚−3}. A book graph Bn (Cm) is a graph obtained from cycle graphs 𝐶𝑚
𝑖 by merging edge uv from
each cycle. The vertex set of Bn (Cm) is y
f
(
)
𝑉(𝐵𝑛(𝐶𝑚)) = {𝑢, 𝑣} ∪ {𝑥𝑖,𝑗∶𝑖= 1, … , 𝑛, 𝑗= 1, … , 𝑚−2 }, and the edge set is
𝐸(𝐵𝑛(𝐶𝑚)) = {𝑢𝑣} ∪ {𝑢𝑥𝑖1 , 𝑥𝑖,𝑗𝑥𝑖,𝑗+1, 𝑥𝑚−2𝑣∶𝑖= 1, … , 𝑛 , 𝑗= 1, … , 𝑚−2}. f
𝑉(𝐵𝑛(𝐶𝑚)) = {𝑢, 𝑣} ∪ {𝑥𝑖,𝑗∶𝑖= 1, … , 𝑛, 𝑗= 1, … , 𝑚−2 }, and the edge set is
𝐸(𝐵𝑛(𝐶𝑚)) = {𝑢𝑣} ∪ {𝑢𝑥𝑖1 , 𝑥𝑖,𝑗𝑥𝑖,𝑗+1, 𝑥𝑚−2𝑣∶𝑖= 1, … , 𝑛 , 𝑗= 1, … , 𝑚−2}. 𝑉(𝐵𝑛(𝐶𝑚)) = {𝑢, 𝑣} ∪ {𝑥𝑖,𝑗∶𝑖= 1, … , 𝑛, 𝑗= 1, … , 𝑚−2 }, and the edge set is
𝐸(𝐵𝑛(𝐶𝑚)) = {𝑢𝑣} ∪ {𝑢𝑥𝑖1 , 𝑥𝑖,𝑗𝑥𝑖,𝑗+1, 𝑥𝑚−2𝑣∶𝑖= 1, … , 𝑛 , 𝑗= 1, … , 𝑚−2}. Furthermore, we define quadruplet book graphs. Furthermore, we define quadruplet book graphs. Definition 2.2. Let 𝐵𝑛
𝑞(𝐶𝑚), 1 ≤𝑞≤4 be the 𝑞𝑡ℎ copy of book graph Bn (Cm) as defined in
Definition 2.1. Let the vertices of 𝐵𝑛
𝑞(𝐶𝑚) be Definition 2.2. Let 𝐵𝑛
𝑞(𝐶𝑚), 1 ≤𝑞≤4 be the 𝑞𝑡ℎ copy of book graph Bn (Cm) as defined in
Definition 2.1. Let the vertices of 𝐵𝑛
𝑞(𝐶𝑚) be 𝑉(𝐵𝑛
𝑞(𝐶𝑚)) = {𝑢𝑞, 𝑣𝑞} ∪ {𝑥𝑖,𝑗
𝑞∶𝑖= 1, … , 𝑛; 𝑗= 1, … , 𝑚−2 }. 𝑉(𝐵𝑛
𝑞(𝐶𝑚)) = {𝑢𝑞, 𝑣𝑞} ∪ {𝑥𝑖,𝑗
𝑞∶𝑖= 1, … , 𝑛; 𝑗= 1, … , 𝑚−2 }. A quadruplet book graph 4𝐵𝑛(𝐶𝑚) is a graph obtained from four copies of book graph
𝐵𝑛
𝑞(𝐶𝑚) by identifying vertex 𝑣𝑞 from book graph 𝐵𝑛
𝑞(𝐶𝑚) as vertex 𝑢𝑞+1 from book graph
𝐵𝑛
𝑞+1(𝐶𝑚) and renaming this vertex as 𝑧𝑞 , 1 ≤𝑞≤3. The vertex set of 5Bn(Cm) is 2 Main result and section 2.2,
respectively. 2 2 https://doi.org/10.1051/itmconf/20213603004 ITM Web of Conferences 36, 03004 (2021)
ICMSA 2020 We determine the tes quadruplet book graphs as described in Theorem 2.4. We determine the tes quadruplet book graphs as described in Theorem 2.4. Theorem 2.4. Let 4𝐵𝑛(𝐶𝑚) be a quadruplet book graph. Then the tes of 4𝐵𝑛(𝐶𝑚) is
⌈
4(𝑚−1)𝑛+6
3
⌉. Proof. Proof. A quadruplet book graph 4𝐵𝑛(𝐶𝑚) has 𝑚 sides and 4𝑛 sheets; therefore, from Definition 2.2. is obtained |𝐸( 4𝐵𝑛(𝐶𝑚))| = 4(𝑚−1)𝑛+ 4. By lower bound in [2] (i.e., 𝑡𝑒𝑠(𝐺) ≥
𝑚𝑎𝑥{⌈
|𝐸|+2
3 ⌉, ⌈
∆(𝐺)+1
2
⌉}), we obtain that 𝑡𝑒𝑠(4𝐵𝑛(𝐶𝑚)) ≥ ⌈
4(𝑚−1)𝑛+6
3
⌉ because any book
graph 4(𝐵𝑛(𝐶𝑚) has a maximum degree ∆(4(𝐵𝑛(𝐶𝑚)) = 2(𝑛+ 1). By constructing an edge
irregular total k-labelling for 4𝐵𝑛(𝐶𝑚) with 𝑘4 = ⌈
4(𝑚−1)𝑛+6
3
⌉ we can prove the upper bound. We know from Definition 2.2. that 𝑧1 = 𝑣1 = 𝑢2, 𝑧2 = 𝑣2 = 𝑢3, 𝑧3 = 𝑣3 = 𝑢4 and we
define the vertex labelling 𝑓 in the following way:
1 irregular total k-labelling for 4𝐵𝑛(𝐶𝑚) with 𝑘4 = ⌈
(
)
3
⌉ we can prove the upper bound. We know from Definition 2.2. that 𝑧1 = 𝑣1 = 𝑢2, 𝑧2 = 𝑣2 = 𝑢3, 𝑧3 = 𝑣3 = 𝑢4 and we
define the vertex labelling 𝑓 in the following way:
(
1) 3
We know from Definition 2.2. that 𝑧1 = 𝑣1 = 𝑢2, 𝑧2 = 𝑣2 = 𝑢3, 𝑧3 = 𝑣3 = 𝑢4 and we
define the vertex labelling 𝑓 in the following way: 𝑓(𝑢1) = 𝑘0 ,
𝑓(𝑣𝑞) = 𝑘𝑞 , 1 ≤𝑞≤4
𝑓(𝑥𝑖,𝑗
𝑞) = 𝑘𝑞−1 + 𝑖+ 𝑗−2, 1 ≤𝑖≤𝑛, 1 ≤𝑗≤𝑦
𝑓(𝑥𝑖,𝑗
𝑞) = 𝑘𝑞−1 +
3𝑗−𝑚+𝒂
6
𝑛+
𝑚−𝑗−𝑏
2
+ 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑦+ 𝑝, 𝑝= 1, 3, … , 2𝑦−5
𝑓(𝑥𝑖,𝑗
𝑞) = 𝑘𝑞−1 +
3𝑗−𝑚−𝑐
6
𝑛+
𝑚−𝑗−𝑑
2
+ 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑦+ 𝑝, 𝑝= 2, 4, … , 2𝑦−𝑒
𝑓(𝑥𝑖,𝑗
𝑞) = 𝑘𝑞−1 +
3𝑗−𝑚+3
6
𝑛+ 𝑖, 𝑖𝑓 𝑚= 0(𝑚𝑜𝑑 3), 1 ≤ 𝑖 ≤𝑟𝑞, 𝑗= 𝑚−3
𝑓(𝑥𝑖,𝑗
𝑞) = 𝑘𝑞, 𝑖𝑓 𝑚= 0(𝑚𝑜𝑑 3), 𝑟𝑞+ 1 ≤ 𝑖≤𝑛, 𝑗= 𝑚−3
𝑓(𝑥𝑖,𝑚−2
𝑞
) = 𝑘𝑞, 1 ≤ 𝑖 ≤𝑛, 1 ≤𝑞≤4
1 ≤𝑖≤𝑛, 1 ≤𝑗≤𝑦 with 𝑘0 = 1, 𝑘𝑞= ⌈
𝑞((𝑚−1)𝑛+1)+2
3
⌉, 𝑟𝑞= 𝑘𝑞−
𝑚−3
3 𝑛+ 𝑘𝑞−1 and 𝑞= 1 for the first book,
𝑞= 2 for the second book, 𝑞= 3 for the third book, 𝑞= 4 for the fourth book and
i. 𝑎= 3, 𝑏= 5, 𝑐= 0, 𝑑= 2, 𝑒= 4, 𝑦=
𝑚
3 for 𝑚= 0(𝑚𝑜𝑑 3),
ii. 𝑎= 1, 𝑏= 5, 𝑐= 2, 𝑑= 2, 𝑒= 6, 𝑦=
𝑚+2
3 for 𝑚= 1(𝑚𝑜𝑑 3),
iii. Proof. 𝑎= 2, 𝑏= 6, 𝑐= 1, 𝑑= 3, 𝑒= 6, 𝑦=
𝑚+1
3 for 𝑚= 2(𝑚𝑜𝑑 3). 𝑞
, 𝑞
, 𝑞
i. 𝑎= 3, 𝑏= 5, 𝑐= 0, 𝑑= 2, 𝑒= 4, 𝑦=
𝑚
3 for 𝑚= 0(𝑚𝑜𝑑 3),
ii. 𝑎= 1, 𝑏= 5, 𝑐= 2, 𝑑= 2, 𝑒= 6, 𝑦=
𝑚+2
3 for 𝑚= 1(𝑚𝑜𝑑 3),
iii. 𝑎= 2, 𝑏= 6, 𝑐= 1, 𝑑= 3, 𝑒= 6, 𝑦=
𝑚+1
3 for 𝑚= 2(𝑚𝑜𝑑 3). We have defined the edge labelling 𝑓 for the first book, the second book and the third
book in [20]. We prove the edge labelling 𝑓 of the fourth book for three different cases. 1. For 𝒎≡𝟎 𝒎𝒐𝒅 𝟑. 1. For 𝒎≡𝟎 𝒎𝒐𝒅 𝟑. 1. For 𝒎≡𝟎 𝒎𝒐𝒅 𝟑. 1. For 𝒎≡𝟎 𝒎𝒐𝒅 𝟑. 3
with 1 ≤𝑖≤𝑛. 2. For 𝒎≡𝟏 𝒎𝒐𝒅 𝟑. 1. For 𝒎≡𝟎 𝒎𝒐𝒅 𝟑. The edge labelling 𝑓 for the fourth book is defined as follows: 𝑓(𝑢4, 𝑣4) =
{
𝑘3 + 𝑛−(
2𝑛+2
3 ) , 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘3 + 𝑛−(
2𝑛
3 ) , 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘3 + 𝑛−(
2𝑛+1
3 ) , 𝑛= 1 (𝑚𝑜𝑑 3) 𝑓(𝑢4𝑥𝑖,1
4 ) = 𝑘3, 1 ≤𝑖≤𝑛
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 𝑘3 + 𝑗𝑛− 2𝑗+ 2 −𝑖, 1 ≤𝑖≤𝑛, 1 ≤𝑗≤
𝑚−3
3
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 𝑘3 +
𝑚−3
3 𝑛−𝑚+ 𝑗+ 5 −𝑖, 1 ≤𝑖≤𝑛, 𝑗=
𝑚
3
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 𝑘3 +
𝑚−3
3 𝑛−𝑚+ 𝑗+ 4 −𝑖, 1 ≤𝑖≤𝑛,
𝑚+3
3
≤𝑗≤𝑚−5 3 https://doi.org/10.1051/itmconf/20213603004 ITM Web of Conferences 36, 03004 (2021) ITM Web of Conferences 36, 03004 (2021)
ICMSA 2020 ICMSA 2020 𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 𝑘3 +
𝑚−3
3 𝑛−𝑖+ 1, 1 ≤𝑖≤ 𝑟4, 𝑗= 𝑚−4
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 2𝑘3 −𝑘4 + 2
𝑚−3
3 𝑛, 𝑟4 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−4
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) =
{
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛+2)
3
, 1 ≤𝑖≤𝑟4, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛)
3 , 1 ≤𝑖≤𝑟4, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛+1)
3
, 1 ≤𝑖≤𝑟4, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) =
{
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛+2)
3
+ 𝑝, 𝑟4 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛)
3 + 𝑝, 𝑟4 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛+1)
3
+ 𝑝, 𝑟4 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
with 1 ≤𝑝≤𝑛−𝑟4 and 𝑟4 = 𝑘4 −
(𝑚−3)
3
𝑛+ 𝑘3
𝑓(𝑥𝑖,𝑗
4 𝑣4) =
{
𝑘3 +
𝑚−6
3 𝑛−2 (
𝑛+1
3 ) + 𝑖−2, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−6
3 𝑛−2
(𝑛+3)
3
+ 𝑖−2, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−6
3 𝑛−2
(𝑛+2)
3
+ 𝑖−2, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 1 (𝑚𝑜𝑑 3) 𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 𝑘3 +
𝑚−3
3 𝑛−𝑖+ 1,
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 2𝑘3 −𝑘4 + 2
𝑚−3
3 𝑛,
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) =
{
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛+2)
3
,
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛)
3 ,
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛+1)
3
,
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) =
{
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛+2)
3
+ 𝑝,
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛)
3 + 𝑝,
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛+1)
3
+ 𝑝, 𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 𝑘3 +
𝑚−3
3 𝑛−𝑖+ 1, 1 ≤𝑖≤ 𝑟4, 𝑗= 𝑚−4
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 2𝑘3 −𝑘4 + 2
𝑚−3
3 𝑛, 𝑟4 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−4
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) =
{
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛+2)
3
, 1 ≤𝑖≤𝑟4, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛)
3 , 1 ≤𝑖≤𝑟4, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛+1)
3
, 1 ≤𝑖≤𝑟4, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) =
{
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛+2)
3
+ 𝑝, 𝑟4 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛)
3 + 𝑝, 𝑟4 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−3
3 𝑛−
(2𝑛+1)
3
+ 𝑝, 𝑟4 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
with 1 ≤𝑝≤𝑛−𝑟4 and 𝑟4 = 𝑘4 −
(𝑚−3)
3
𝑛+ 𝑘3
𝑓(𝑥𝑖,𝑗
4 𝑣4) =
{
𝑘3 +
𝑚−6
3 𝑛−2 (
𝑛+1
3 ) + 𝑖−2, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−6
3 𝑛−2
(𝑛+3)
3
+ 𝑖−2, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−6
3 𝑛−2
(𝑛+2)
3
+ 𝑖−2, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 1 (𝑚𝑜𝑑 3) with 1 ≤𝑝≤𝑛
𝑟4 and 𝑟4
𝑘4
3
𝑛+ 𝑘3
𝑓(𝑥𝑖,𝑗
4 𝑣4) =
{
𝑘3 +
𝑚−6
3 𝑛−2 (
𝑛+1
3 ) + 𝑖−2, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−6
3 𝑛−2
(𝑛+3)
3
+ 𝑖−2, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘3 +
𝑚−6
3 𝑛−2
(𝑛+2)
3
+ 𝑖−2, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 1 (𝑚𝑜𝑑 3) Under the labelling 𝑓, we have the weight of the edges of the fourth book graph as below:
2 ) =
{
2𝑘3 + 𝑘4 + (
𝑛−2
3 ) , 𝑛= 2 (𝑚𝑜𝑑 3)
2𝑘3 + 𝑘4 + (
𝑛
3) , 𝑛= 0 (𝑚𝑜𝑑 3)
2𝑘3 + 𝑘4 + (
𝑛−1
3 ) , 𝑛= 1 (𝑚𝑜𝑑 3) 𝜔𝑓 (𝑢4𝑥𝑖,1
4 ) = 3𝑘3 + 𝑖−1,
𝜔𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 3𝑘3 + 𝑗𝑛+ 𝑖−1, 1 ≤𝑗≤
𝑚−3
3
𝜔𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 3𝑘3 + 𝑗𝑛+ 𝑖,
𝑚
3 ≤𝑗≤𝑚−4
𝜔𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) =
{
2𝑘3 + 𝑘4 + 𝑖+
(2𝑚−8)𝑛−1
3
, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
2𝑘3 + 𝑘4 + 𝑖+
(2𝑚−8)𝑛
3
, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
2𝑘3 + 𝑘4 + 𝑖+
(2𝑚−8)𝑛−2
3
, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
𝜔𝑓(𝑥𝑖,𝑗
4 𝑣4) =
{
𝑘3 + 2𝑘4 + 𝑖+
(2𝑚−4)𝑛−2
3
, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘3 + 2𝑘4 + 𝑖+
(2𝑚−4)𝑛
3
, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘3 + 2𝑘4 + 𝑖+
(2𝑚−4)𝑛−1
3
, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3) 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3) {
with 1 ≤𝑖≤𝑛. 2. For 𝒎≡𝟏 𝒎𝒐𝒅 𝟑. The edge labelling 𝑓 for the fourth book is defined as follows:
(
4
4) 𝑓(𝑢4, 𝑣4) = 𝑘3 + 𝑛,
𝑓(𝑢4𝑥𝑖,1
4 ) = 𝑘3, 1 ≤𝑖≤𝑛
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 𝑘3 + 𝑗𝑛− 2𝑗−𝑖+ 2, 1 ≤𝑖≤𝑛, 1 ≤𝑗≤
𝑚−1
3
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 𝑘3 +
𝑚−1
3 𝑛+ 𝑗−𝑚−𝑖+ 4, 1 ≤𝑖≤𝑛,
𝑚+2
3
≤𝑗≤𝑚−4
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 𝑘3 +
𝑚−4
3 𝑛+ 1, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3 https://doi.org/10.1051/itmconf/20213603004 ITM Web of Conferences 36, 03004 (2021) ITM Web of Conferences 36, 03004 (2021)
ICMSA 2020 𝑓(𝑥𝑖,𝑗
4 𝑣4) = 𝑘3 +
𝑚−4
3 𝑛+ 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2. 𝑓(𝑥𝑖,𝑗
4 𝑣4) = 𝑘3 +
𝑚−4
3 𝑛+ 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2. Under the labelling 𝑓, we have the weight of the edges of the fourth book graph as below:
4 Under the labelling 𝑓, we have the weight of the edges of the fourth book graph as below
4
4 𝜔𝑓(𝑢4𝑥𝑖,1
4 ) = 3𝑘3 + 𝑖−1,
𝜔𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 3𝑘3 + 𝑗𝑛+ 𝑖−1, 1 ≤𝑗≤
𝑚−1
3
𝜔𝑓(𝑢4, 𝑣4) = 2𝑘3 + 𝑘4 + 𝑛,
𝜔𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 3𝑘3 + 𝑗𝑛+ 𝑖,
𝑚+2
3
≤𝑗≤𝑚−4
𝜔𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 2𝑘3 + 𝑘4 + 2 (
𝑚−4
3 ) 𝑛+ 𝑖, 𝑗= 𝑚−3
𝜔𝑓(𝑥𝑖,𝑗
4 𝑣4) = 𝑘3 + 2𝑘4 +
𝑚−4
3 𝑛+ 𝑖, 𝑗= 𝑚−2,
with 1 ≤𝑖≤𝑛. 1 ≤𝑗≤
𝑚−1
3 3. For 𝒎≡𝟐 𝒎𝒐𝒅 𝟑. The edge labelling 𝑓 for the fourth book is defined as below: 𝑓(𝑢4, 𝑣4) =
{
𝑘3 + 𝑛−(
𝑛+1
3 ) , 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘3 + 𝑛−(
𝑛
3) , 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘3 + 𝑛−(
𝑛+2
3 ) , 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 𝑘3 + 𝑗𝑛−2𝑗−𝑖+ 2, 1 ≤𝑖≤𝑛, 1 ≤𝑗≤
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 𝑘3 +
𝑚−2
3 𝑛+ 𝑗−𝑚−𝑖+ 5, 1 ≤𝑖≤𝑛,
𝑚+1
3
≤
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) =
{
𝑘3 +
𝑚−2
3 𝑛−(
𝑛+1
3 ) , 1 ≤𝑖≤𝑛, 𝑗= 𝑚−
𝑘3 +
𝑚−2
3 𝑛−(
𝑛
3) , 1 ≤𝑖≤𝑛, 𝑗= 𝑚−
𝑘3 +
𝑚−2
3 𝑛−(
𝑛+2
3 ) , 1 ≤𝑖≤𝑛, 𝑗= 𝑚−
𝑓(𝑥𝑖,𝑗
4 𝑣4) =
{
𝑘3 +
𝑚−2
3 𝑛−(
𝑛+1
3 ) + 𝑖, 𝑗= 𝑚−2, 𝑛= 2 (
𝑘3 +
𝑚−2
3 𝑛−(
𝑛
3) + 𝑖, 𝑗= 𝑚−2, 𝑛= 0 (
𝑘3 +
𝑚−2
3 𝑛−(
𝑛+2
3 ) + 𝑖, 𝑗= 𝑚−2, 𝑛= 1 (𝑚 𝑓(𝑢4, 𝑣4) =
{
𝑘3 + 𝑛−(
𝑛+1
3 ) ,
𝑘3 + 𝑛−(
𝑛
3) ,
𝑘3 + 𝑛−(
𝑛+2
3 ) ,
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 𝑘3 + 𝑗𝑛−2𝑗−𝑖+ 2,
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 𝑘3 +
𝑚−2
3 𝑛+ 𝑗−𝑚−𝑖+ 5,
𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) =
{
𝑘3 +
𝑚−2
3 𝑛−(
𝑛+1
3 ) ,
𝑘3 +
𝑚−2
3 𝑛−(
𝑛
3) ,
𝑘3 +
𝑚−2
3 𝑛−(
𝑛+2
3 ) ,
𝑓(𝑥𝑖,𝑗
4 𝑣4) =
{
𝑘3 +
𝑚−2
3 𝑛−(
𝑛+1
3 ) + 𝑖,
𝑘3 +
𝑚−2
3 𝑛−(
𝑛
3) + 𝑖,
𝑘3 +
𝑚−2
3 𝑛−(
𝑛+2
3 ) + 𝑖, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑛= 0 (𝑚𝑜𝑑 3)
𝑛= 1 (𝑚𝑜𝑑 3) 𝑛= 1 (𝑚𝑜𝑑 3) 1 ≤𝑖≤𝑛, 1 ≤𝑗≤
𝑚−2
3
1 ≤𝑖≤𝑛,
𝑚+1
3
≤𝑗≤𝑚−4
1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑗= 𝑚−2, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑗= 𝑚−2, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑗= 𝑚−2, 𝑛= 1 (𝑚𝑜𝑑 3). 3. For 𝒎≡𝟐 𝒎𝒐𝒅 𝟑. Under the labelling 𝑓, we have the weight of the edges of the fourth book graph as below
(
4
4 ) 𝜔𝑓 (𝑢4𝑥𝑖,1
4 ) = 3𝑘3 + 𝑖−1,
𝜔𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 3𝑘3 + 𝑗𝑛+ 𝑖−1,
𝜔𝑓(𝑢4, 𝑣4) =
{
2𝑘3 + 𝑘4 + 𝑛−(
𝑛+1
3 ) ,
2𝑘3 + 𝑘4 + 𝑛−(
𝑛
3) ,
2𝑘3 + 𝑘4 + 𝑛−(
𝑛+2
3 ) ,
𝜔𝑓(𝑥𝑖𝑗
4 𝑥𝑖𝑗+1
4
) = 3𝑘3 + 𝑗𝑛+ 𝑖 𝜔𝑓 (𝑢4𝑥𝑖,1
4 ) = 3𝑘3 + 𝑖−1,
𝜔𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 3𝑘3 + 𝑗𝑛+ 𝑖−1, 1 ≤𝑗≤
𝑚−1
3
𝜔𝑓(𝑢4, 𝑣4) =
{
2𝑘3 + 𝑘4 + 𝑛−(
𝑛+1
3 ) , 𝑛= 2 (𝑚𝑜𝑑 3)
2𝑘3 + 𝑘4 + 𝑛−(
𝑛
3) , 𝑛= 0 (𝑚𝑜𝑑 3)
2𝑘3 + 𝑘4 + 𝑛−(
𝑛+2
3 ) , 𝑛= 1 (𝑚𝑜𝑑 3)
𝜔𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) = 3𝑘3 + 𝑗𝑛+ 𝑖,
𝑚+2
3
≤𝑗≤𝑚−4
𝜔𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) =
{
2𝑘3 + 𝑘4 + 𝑖+
(2𝑚−8)𝑛−1
3
, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
2𝑘3 + 𝑘4 + 𝑖+
(2𝑚−8)𝑛
3
, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
2𝑘3 + 𝑘4 + 𝑖+
(2𝑚−8)𝑛−2
3
, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3) 𝑗
𝑗
𝜔𝑓(𝑥𝑖,𝑗
4 𝑥𝑖,𝑗+1
4
) =
{
2𝑘3 + 𝑘4 + 𝑖+
(2𝑚−8)𝑛−1
3
,
2𝑘3 + 𝑘4 + 𝑖+
(2𝑚−8)𝑛
3
,
2𝑘3 + 𝑘4 + 𝑖+
(2𝑚−8)𝑛−2
3
, 5 5 https://doi.org/10.1051/itmconf/20213603004 ITM Web of Conferences 36, 03004 (2021) ITM Web of Conferences 36, 03004 (2021)
ICMSA 2020 𝜔𝑓(𝑥𝑖,𝑗
4 𝑣4) =
{
𝑘3 + 2𝑘4 + 𝑖+
(2𝑚−4)𝑛−2
3
, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘3 + 2𝑘4 + 𝑖+
(2𝑚−4)𝑛
3
, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘3 + 2𝑘4 + 𝑖+
(2𝑚−4)𝑛−1
3
, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3), {
with 1 ≤𝑖≤𝑛. For Cases 1, 2 and 3, under the labelling 𝑓, the weights of edges of 4𝐵𝑛(𝐶𝑚) constitute the
set {3, … , 4(𝑚−1)𝑛+ 6}. This shows that the weights of all the edges of 4𝐵𝑛(𝐶𝑚) are
different and thus tes (4𝐵𝑛(𝐶𝑚)) = ⌈
4(𝑚−1)𝑛+6
3
⌉. ◼ 2.2 Total edge irregularity strength of quintuplet book graphs In this section we discuss the tes of quintuplet book graphs. Theorem 2.5. Let 5𝐵𝑛(𝐶𝑚) be a quintuplet book graph. Then the tes of (5𝐵𝑛(𝐶𝑚)) is
⌈
5(𝑚−1)𝑛+7
3
⌉. Proof. A quintuplet book graph 5𝐵𝑛(𝐶𝑚) has 𝑚 sides and 5𝑛 sheets; therefore from Definition 2.3. is obtained |𝐸( 5𝐵𝑛(𝐶𝑚))| = 5(𝑚−1)𝑛+ 7. Bača et al. in [2] give the lower bound for
tes(G); that is, tes(G) ≥ 𝑚𝑎𝑥{⌈
|𝐸|+2
3 ⌉, ⌈
∆(𝐺)+1
2
⌉}. Any book graph 5(𝐵𝑛(𝐶𝑚) has maximum
degree ∆(5(𝐵𝑛(𝐶𝑚)) = 2(𝑛+ 1), so we have tes(5𝐵𝑛(𝐶𝑚)) ≥ ⌈
5(𝑚−1)𝑛+7
3
⌉. To prove the
upper bound, we construct an edge irregular total k-labelling for 5𝐵𝑛(𝐶𝑚) with 𝑘5 =
⌈
5(𝑚−1)𝑛+7
3
⌉ for three different cases. 3
We know from Definition 2.3. that 𝑧1 = 𝑣1 = 𝑢2, 𝑧2 = 𝑣2 = 𝑢3, 𝑧3 = 𝑣3 = 𝑢4, 𝑧4 = 𝑣4
= 𝑢5, and we define the vertex labelling 𝑓 similarly as in Theorem 2.4. 3
We know from Definition 2.3. that 𝑧1 = 𝑣1 = 𝑢2, 𝑧2 = 𝑣2 = 𝑢3, 𝑧3 = 𝑣3 = 𝑢4, 𝑧4 = 𝑣4
= 𝑢5, and we define the vertex labelling 𝑓 similarly as in Theorem 2.4. with 𝑞= 1 for the first book, 𝑞= 2 for the second book, 𝑞= 3 for the third book, 𝑞= 4 for
((
1)
1) 2 with 𝑞= 1 for the first book, 𝑞= 2 for the second book, 𝑞= 3 for the third book, 𝑞= 4 for
the fourth book, 𝑞= 5 for the fifth book; we define 𝑘0 = 1, 𝑘𝑞= ⌈
𝑞((𝑚−1)𝑛+1)+2
3
⌉. For the three cases, we define the edge labelling 𝑓 for the fifth book as follows:
1. For 𝒎≡𝟎 𝒎𝒐𝒅 𝟑. 2. For 𝒎≡𝟏 𝒎𝒐𝒅 𝟑. We construct the edge labelling 𝑓 for the fifth book as follows:
5
5 𝑢5, 𝑣5) = 𝑘4 + 𝑛,
𝑢5𝑥𝑖,1
5 ) = 𝑘4 + 1, 𝑓(𝑢5, 𝑣5) = 𝑘4 + 𝑛,
𝑓(𝑢5𝑥𝑖,1
5 ) = 𝑘4 + 1, 1 ≤𝑖≤𝑛
𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) = 𝑘4 + 𝑗𝑛− 2𝑗+ 3 −𝑖, 1 ≤𝑖≤𝑛, 1 ≤𝑗≤
𝑚−1
3
𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) = 𝑘4 +
𝑚−1
3 𝑛+ 𝑗+ 5 −𝑚−𝑖, 1 ≤𝑖≤𝑛,
𝑚+2
3
≤𝑗≤𝑚−4
𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) = 𝑘3 +
𝑚−4
3 𝑛+ 1, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3
𝑓(𝑥𝑖,𝑗
5 𝑣5) = 𝑘3 +
𝑚−4
3 𝑛+ 𝑖−1, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2. 𝑓(𝑢, 𝑣) = 𝑘4 + 𝑛,
𝑓(𝑢5𝑥𝑖,1
5 ) = 𝑘4 + 1, 1 ≤𝑖≤𝑛
𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) = 𝑘4 + 𝑗𝑛− 2𝑗+ 3 −𝑖, 1 ≤𝑖≤𝑛, 1 ≤𝑗≤
𝑚−1
3
𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) = 𝑘4 +
𝑚−1
3 𝑛+ 𝑗+ 5 −𝑚−𝑖, 1 ≤𝑖≤𝑛,
𝑚+2
3
≤𝑗≤𝑚−4
𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) = 𝑘3 +
𝑚−4
3 𝑛+ 1, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3
𝑓(𝑥𝑖,𝑗
5 𝑣5) = 𝑘3 +
𝑚−4
3 𝑛+ 𝑖−1, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2. Under the labelling 𝑓, we have the weights of the edges of the fifth book graph as follow
(
5
5 ) 𝜔𝑓(𝑢5𝑥𝑖,1
5 ) = 3𝑘4 + 𝑖,
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) = 3𝑘4 + 𝑗𝑛+ 𝑖, 1 ≤𝑗≤
𝑚−1
3
𝜔𝑓(𝑢5, 𝑣5) = 2𝑘4 + 𝑘5 + 𝑛,
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) = 3𝑘4 + 𝑗𝑛+ 𝑖+ 1,
𝑚+2
3
≤𝑗≤𝑚−4
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) = 2𝑘4 + 𝑘5 + 2 (
𝑚−4
3 ) 𝑛+ 𝑖, 𝑗= 𝑚−3
𝜔𝑓(𝑥𝑖,𝑗
5 𝑣5) = 𝑘4 + 2𝑘5 +
𝑚−4
3 𝑛+ 𝑖−1, 𝑗= 𝑚−2,
with 1 ≤𝑖≤𝑛. 𝜔𝑓(𝑢5𝑥𝑖,1
5 ) = 3𝑘4 + 𝑖,
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) = 3𝑘4 + 𝑗𝑛+ 𝑖, 1 ≤𝑗≤
𝑚−1
3
𝜔𝑓(𝑢5, 𝑣5) = 2𝑘4 + 𝑘5 + 𝑛,
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) = 3𝑘4 + 𝑗𝑛+ 𝑖+ 1,
𝑚+2
3
≤𝑗≤𝑚−4
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) = 2𝑘4 + 𝑘5 + 2 (
𝑚−4
3 ) 𝑛+ 𝑖, 𝑗= 𝑚−3
𝜔𝑓(𝑥𝑖,𝑗
5 𝑣5) = 𝑘4 + 2𝑘5 +
𝑚−4
3 𝑛+ 𝑖−1, 𝑗= 𝑚−2,
with 1 ≤𝑖≤𝑛. 1 ≤𝑗≤
𝑚−1
3 Proof. {
3
with 1 ≤𝑖≤𝑛. 3
with 1 ≤𝑖≤𝑛. 2. For 𝒎≡𝟏 𝒎𝒐𝒅 𝟑. We construct the edge labelling 𝑓for the fifth book as follows: 2. For 𝒎≡𝟏 𝒎𝒐𝒅 𝟑. Proof. The edge labelling 𝑓 for the fifth book is defined in the following way: 𝑓(𝑢5, 𝑣5) =
{
𝑘4 + 𝑛−(
2𝑛+2
3 ) , 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 + 𝑛−(
2𝑛
3 ) , 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 + 𝑛−(
2𝑛+1
3 ) , 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑤5𝑥𝑖,1
5 ) = {
𝑘4 −1 1 ≤𝑖≤𝑛, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 + 1, 1 ≤𝑖≤𝑛, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4, 1 ≤𝑖≤𝑛, 𝑛= 1 (𝑚𝑜𝑑 3) 𝑓(𝑢5, 𝑣5) =
{
𝑘4 + 𝑛−(
2𝑛+2
3 ) , 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 + 𝑛−(
2𝑛
3 ) , 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 + 𝑛−(
2𝑛+1
3 ) , 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑤5𝑥𝑖,1
5 ) = {
𝑘4 −1 1 ≤𝑖≤𝑛, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 + 1, 1 ≤𝑖≤𝑛, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4, 1 ≤𝑖≤𝑛, 𝑛= 1 (𝑚𝑜𝑑 3) 𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
𝑘4 + 𝑗𝑛−2𝑗−𝑖+ 1, 1 ≤𝑖≤𝑛,
𝑚+3
3
≤𝑗≤𝑚−5, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 + 𝑗𝑛−2𝑗−𝑖+ 3, 1 ≤𝑖≤𝑛,
𝑚+3
3
≤𝑗≤𝑚−5, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 + 𝑗𝑛−2𝑗−𝑖+ 2, 1 ≤𝑖≤𝑛,
𝑚+3
3
≤𝑗≤𝑚−5, 𝑛= 1 (𝑚𝑜𝑑 3) 6 https://doi.org/10.1051/itmconf/20213603004 ITM Web of Conferences 36, 03004 (2021) ITM Web of Conferences 36, 03004 (2021)
ICMSA 2020 𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
𝑘4 +
𝑚−3
3 𝑛−𝑚+ 𝑗−𝑖+ 2, 1 ≤𝑖≤𝑟2, 𝑗= 𝑚−4, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−3
3 𝑛−𝑚+ 𝑗−𝑖+ 4, 1 ≤𝑖≤𝑟2, 𝑗= 𝑚−4, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−3
3 𝑛−𝑚+ 𝑗−𝑖+ 3, 1 ≤𝑖≤𝑟2, 𝑗= 𝑚−4, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
)
{
2𝑘4 −𝑘5 + 2
𝑚−3
3 𝑛−𝑚+ 𝑗+ 2, 𝑟5 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−4, 𝑛= 2 (𝑚𝑜𝑑 3)
2𝑘4 −𝑘5 + 2
𝑚−3
3 𝑛−𝑚+ 𝑗+ 4, 𝑟5 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−4, 𝑛= 0 (𝑚𝑜𝑑 3)
2𝑘4 −𝑘5 + 2
𝑚−3
3 𝑛−𝑚+ 𝑗+ 3, 𝑟5 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−4, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
𝑘4 +
𝑚−3
3 𝑛−
2𝑛+2
3
, 1 ≤𝑖≤𝑟5, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−3
3 𝑛−
2𝑛
3 , 1 ≤𝑖≤𝑟5, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−3
3 𝑛−
2𝑛+1
3
, 1 ≤𝑖≤𝑟5, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
𝑘4 +
𝑚−3
3 𝑛−
(2𝑛+2)
3
+ 𝑝, 𝑟5 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−3
3 𝑛−
(2𝑛)
3 + 𝑝, 𝑟5 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−3
3 𝑛−
(2𝑛+1)
3
+ 𝑝, 𝑟5 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
with 1 ≤𝑝≤𝑛−𝑟5 and 𝑟5 = 𝑘5 −
(𝑚−3)
3
𝑛+ 𝑘4
𝑓(𝑥𝑖,𝑗
5 𝑣5) =
{
𝑘4 +
𝑚−6
3 𝑛+ 2
𝑛+1
3 + 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−6
3 𝑛+ 2
𝑛+3
3 + 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−6
3 𝑛+ 2
𝑛+2
3 + 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 1 (𝑚𝑜𝑑 3) 𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
𝑘4 +
𝑚−3
3 𝑛−𝑚+ 𝑗−𝑖+ 2, 1 ≤𝑖≤𝑟2, 𝑗= 𝑚−4, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−3
3 𝑛−𝑚+ 𝑗−𝑖+ 4, 1 ≤𝑖≤𝑟2, 𝑗= 𝑚−4, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−3
3 𝑛−𝑚+ 𝑗−𝑖+ 3, 1 ≤𝑖≤𝑟2, 𝑗= 𝑚−4, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
)
{
2𝑘4 −𝑘5 + 2
𝑚−3
3 𝑛−𝑚+ 𝑗+ 2, 𝑟5 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−4, 𝑛= 2 (𝑚𝑜𝑑 3)
2𝑘4 −𝑘5 + 2
𝑚−3
3 𝑛−𝑚+ 𝑗+ 4, 𝑟5 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−4, 𝑛= 0 (𝑚𝑜𝑑 3)
2𝑘4 −𝑘5 + 2
𝑚−3
3 𝑛−𝑚+ 𝑗+ 3, 𝑟5 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−4, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
𝑘4 +
𝑚−3
3 𝑛−
2𝑛+2
3
, 1 ≤𝑖≤𝑟5, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−3
3 𝑛−
2𝑛
3 , 1 ≤𝑖≤𝑟5, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−3
3 𝑛−
2𝑛+1
3
, 1 ≤𝑖≤𝑟5, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
𝑘4 +
𝑚−3
3 𝑛−
(2𝑛+2)
3
+ 𝑝, 𝑟5 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−3
3 𝑛−
(2𝑛)
3 + 𝑝, 𝑟5 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−3
3 𝑛−
(2𝑛+1)
3
+ 𝑝, 𝑟5 + 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
with 1 ≤𝑝≤𝑛−𝑟5 and 𝑟5 = 𝑘5 −
(𝑚−3)
3
𝑛+ 𝑘4
𝑓(𝑥𝑖,𝑗
5 𝑣5) =
{
𝑘4 +
𝑚−6
3 𝑛+ 2
𝑛+1
3 + 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−6
3 𝑛+ 2
𝑛+3
3 + 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−6
3 𝑛+ 2
𝑛+2
3 + 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 1 (𝑚𝑜𝑑 3) with 1 ≤𝑝≤𝑛−𝑟5 and 𝑟5 = 𝑘5 −
3
𝑛+ 𝑘4
𝑓(𝑥𝑖,𝑗
5 𝑣5) =
{
𝑘4 +
𝑚−6
3 𝑛+ 2
𝑛+1
3 + 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−6
3 𝑛+ 2
𝑛+3
3 + 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−6
3 𝑛+ 2
𝑛+2
3 + 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 1 (𝑚𝑜𝑑 3) 𝑓(𝑥𝑖,𝑗
5 𝑣5) =
{
𝑘4 +
𝑚−6
3 𝑛+ 2
𝑛+1
3 + 𝑖,
𝑘4 +
𝑚−6
3 𝑛+ 2
𝑛+3
3 + 𝑖,
𝑘4 +
𝑚−6
3 𝑛+ 2
𝑛+2
3 + 𝑖, Under the labelling 𝑓, we have the weights of the edges of the fifth book graph as follows: g 𝑓,
g
g
g p
𝜔𝑓 (𝑢5𝑥𝑖,1
5 ) = {
3𝑘4 + 𝑖−2, 𝑛= 2 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑖, 𝑛= 0 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑖−1, 𝑛= 1 (𝑚𝑜𝑑 3)
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
3𝑘4 + 𝑗𝑛+ 𝑖−2, 1 ≤𝑗≤
𝑚−2
3 , 𝑛= 2 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑗𝑛+ 𝑖, 1 ≤𝑗≤
𝑚−2
3 , 𝑛= 0 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑗𝑛+ 𝑖−1, 1 ≤𝑗≤
𝑚−2
3 , 𝑛= 1 (𝑚𝑜𝑑 3)
𝜔𝑓(𝑢5, 𝑣5) =
{
2𝑘4 + 𝑘5 −(
𝑛−2
3 ) , 𝑛= 2 (𝑚𝑜𝑑 3)
2𝑘4 + 𝑘5 −(
𝑛
3) , 𝑛= 0 (𝑚𝑜𝑑 3)
2𝑘4 + 𝑘5 −(
𝑛−1
3 ) , 𝑛= 1 (𝑚𝑜𝑑 3)
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
3𝑘4 + 𝑗𝑛+ 𝑖−1,
𝑚
3 ≤𝑗≤𝑚−4, 𝑛= 2 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑗𝑛+ 𝑖+ 1,
𝑚
3 ≤𝑗≤𝑚−4, 𝑛= 0 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑗𝑛+ 𝑖,
𝑚
3 ≤𝑗≤𝑚−4, 𝑛= 1 (𝑚𝑜𝑑 3)
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛−2
3
, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛
3
, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛−1
3
, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3) g 𝑓
g
g
g p
𝜔𝑓 (𝑢5𝑥𝑖,1
5 ) = {
3𝑘4 + 𝑖−2, 𝑛= 2 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑖, 𝑛= 0 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑖−1, 𝑛= 1 (𝑚𝑜𝑑 3)
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
3𝑘4 + 𝑗𝑛+ 𝑖−2, 1 ≤𝑗≤
𝑚−2
3 , 𝑛= 2 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑗𝑛+ 𝑖, 1 ≤𝑗≤
𝑚−2
3 , 𝑛= 0 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑗𝑛+ 𝑖−1, 1 ≤𝑗≤
𝑚−2
3 , 𝑛= 1 (𝑚𝑜𝑑 3)
𝜔𝑓(𝑢5, 𝑣5) =
{
2𝑘4 + 𝑘5 −(
𝑛−2
3 ) , 𝑛= 2 (𝑚𝑜𝑑 3)
2𝑘4 + 𝑘5 −(
𝑛
3) , 𝑛= 0 (𝑚𝑜𝑑 3)
2𝑘4 + 𝑘5 −(
𝑛−1
3 ) , 𝑛= 1 (𝑚𝑜𝑑 3)
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
3𝑘4 + 𝑗𝑛+ 𝑖−1,
𝑚
3 ≤𝑗≤𝑚−4, 𝑛= 2 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑗𝑛+ 𝑖+ 1,
𝑚
3 ≤𝑗≤𝑚−4, 𝑛= 0 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑗𝑛+ 𝑖,
𝑚
3 ≤𝑗≤𝑚−4, 𝑛= 1 (𝑚𝑜𝑑 3)
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛−2
3
, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛
3
, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛−1
3
, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3) 𝑛= 2 (𝑚𝑜𝑑 3)
𝑛= 0 (𝑚𝑜𝑑 3)
𝑛= 1 (𝑚𝑜𝑑 3) 1 ≤𝑗≤
𝑚−2
3 , 𝑛= 2 (𝑚𝑜𝑑 3)
1 ≤𝑗≤
𝑚−2
3 , 𝑛= 0 (𝑚𝑜𝑑 3)
1 ≤𝑗≤
𝑚−2
3 , 𝑛= 1 (𝑚𝑜𝑑 3)
𝑛= 2 (𝑚𝑜𝑑 3)
𝑛= 0 (𝑚𝑜𝑑 3)
𝑛= 1 (𝑚𝑜𝑑 3) 3
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
3𝑘4 + 𝑗𝑛+ 𝑖−1,
𝑚
3 ≤𝑗≤𝑚−4, 𝑛= 2 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑗𝑛+ 𝑖+ 1,
𝑚
3 ≤𝑗≤𝑚−4, 𝑛= 0 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑗𝑛+ 𝑖,
𝑚
3 ≤𝑗≤𝑚−4, 𝑛= 1 (𝑚𝑜𝑑 3)
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛−2
3
, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛
3
, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛−1
3
, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3) 3
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
3𝑘4 + 𝑗𝑛+ 𝑖−1,
𝑚
3 ≤𝑗≤𝑚−4, 𝑛= 2 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑗𝑛+ 𝑖+ 1,
𝑚
3 ≤𝑗≤𝑚−4, 𝑛= 0 (𝑚𝑜𝑑 3)
3𝑘4 + 𝑗𝑛+ 𝑖,
𝑚
3 ≤𝑗≤𝑚−4, 𝑛= 1 (𝑚𝑜𝑑 3)
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛−2
3
, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛
3
, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛−1
3
, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3) 7 https://doi.org/10.1051/itmconf/20213603004 ITM Web of Conferences 36, 03004 (2021) ITM Web of Conferences 36, 03004 (2021)
ICMSA 2020 ICMSA 2020 𝜔𝑓(𝑥𝑖,𝑗
5 𝑣5) =
{
𝑘4 + 2𝑘5 + 𝑖+
(𝑚−4)𝑛−2
3
, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 + 2𝑘5 + 𝑖+
(𝑚−4)𝑛
3
, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 + 2𝑘5 + 𝑖+
(𝑚−4)𝑛−4
3
, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3), {
with 1 ≤𝑖≤𝑛. 3. For 𝒎≡𝟐 𝒎𝒐𝒅 𝟑. 𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
𝑘4 +
𝑚−2
3 𝑛−(
𝑛+1
3 ) ,
𝑘4 +
𝑚−2
3 𝑛−(
𝑛
3) ,
𝑘4 +
𝑚−2
3 𝑛−(
𝑛+2
3 ) ,
𝑓(𝑥𝑖,𝑗
5 𝑣5) =
{
𝑘4 +
𝑚−2
3 𝑛−2 (
𝑛+1
3 ) + 𝑖,
𝑘4 +
𝑚−2
3 𝑛−2 (
𝑛
3) + 𝑖,
𝑘4 +
𝑚−2
3 𝑛−2 (
𝑛+2
3 ) + 𝑖, 𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
𝑘4 +
𝑚−2
3 𝑛−(
𝑛+1
3 ) , 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−2
3 𝑛−(
𝑛
3) , 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−2
3 𝑛−(
𝑛+2
3 ) , 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑥𝑖,𝑗
5 𝑣5) =
{
𝑘4 +
𝑚−2
3 𝑛−2 (
𝑛+1
3 ) + 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−2
3 𝑛−2 (
𝑛
3) + 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−2
3 𝑛−2 (
𝑛+2
3 ) + 𝑖, 1 ≤𝑖≤𝑛 𝑗= 𝑚−2, 𝑛= 1 (𝑚𝑜𝑑 3). 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 2 (𝑚𝑜𝑑 3)
1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 0 (𝑚𝑜𝑑 3)
1 ≤𝑖≤𝑛 𝑗= 𝑚−2, 𝑛= 1 (𝑚𝑜𝑑 3). Under the labelling 𝑓, the weights of edges of 5𝐵𝑛(𝐶𝑚) for cases 1, 2 and 3 constitute the
set {3, … , 5(𝑚−1)𝑛+ 7}. This shows that the weights of all the edges of (5𝐵𝑛(𝐶𝑚) are
(
) 3. For 𝒎≡𝟐 𝒎𝒐𝒅 𝟑. We define the edge labelling 𝑓 for the fifth book as below: 𝑓(𝑢5, 𝑣5) =
{
𝑘4 + 𝑛−(
𝑛+1
3 ) , 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 + 𝑛−(
𝑛
3) , 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 + 𝑛−(
𝑛+2
3 ) , 𝑛= 1 (𝑚𝑜𝑑 3) 1 ≤𝑖≤𝑛, 𝑛= 2 (𝑚𝑜𝑑 3)
1 ≤𝑖≤𝑛, 𝑛= 0 (𝑚𝑜𝑑 3)
1 ≤𝑖≤𝑛, 𝑛= 1 (𝑚𝑜𝑑 3)
2 𝑓(𝑢5𝑥𝑖,1
5 ) = {
𝑘4,
𝑘4 + 1,
𝑘4 −1, 𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
𝑘4 + 𝑗𝑛−2𝑗−𝑖+ 2, 1 ≤𝑖≤𝑛, 1 ≤𝑗≤
𝑚−2
3 , 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 + 𝑗𝑛−2𝑗−𝑖+ 3, 1 ≤𝑖≤𝑛, 1 ≤𝑗≤
𝑚−2
3 , 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 + 𝑗𝑛−2𝑗−𝑖+ 1, 1 ≤𝑖≤𝑛, 1 ≤𝑗≤
𝑚−2
3 , 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
𝑘4 +
𝑚−2
3 𝑛+ 𝑗+ 5 −𝑚−𝑖,
𝑚+1
3
≤𝑗≤𝑚−4, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−2
3 𝑛+ 𝑗+ 6 −𝑚−𝑖,
𝑚+1
3
≤𝑗≤𝑚−4, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−2
3 𝑛+ 𝑗+ 4 −𝑚−𝑖,
𝑚+1
3
≤𝑗≤𝑚−4, 𝑛= 1 (𝑚𝑜𝑑 3) 8 https://doi.org/10.1051/itmconf/20213603004 ITM Web of Conferences 36, 03004 (2021) ITM Web of Conferences 36, 03004 (2021)
ICMSA 2020 𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
𝑘4 +
𝑚−2
3 𝑛−(
𝑛+1
3 ) , 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−2
3 𝑛−(
𝑛
3) , 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−2
3 𝑛−(
𝑛+2
3 ) , 1 ≤𝑖≤𝑛, 𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑓(𝑥𝑖,𝑗
5 𝑣5) =
{
𝑘4 +
𝑚−2
3 𝑛−2 (
𝑛+1
3 ) + 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−2
3 𝑛−2 (
𝑛
3) + 𝑖, 1 ≤𝑖≤𝑛, 𝑗= 𝑚−2, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑘4 +
𝑚−2
3 𝑛−2 (
𝑛+2
3 ) + 𝑖, 1 ≤𝑖≤𝑛 𝑗= 𝑚−2, 𝑛= 1 (𝑚𝑜𝑑 3). 3 Conclusion We have constructed an edge irregular total k-labelling for the fourth book and the fifth book. We found that the total edge irregularity strength of quadruplet book graphs is tes 4𝐵𝑛(𝐶𝑚) =
⌈
4(𝑚−1)𝑛+6
3
⌉ and of quintuplet book graphs is tes 5𝐵𝑛(𝐶𝑚) = ⌈
5(𝑚−1)𝑛+7
3
⌉. Based on the labelling, we see that there is a similarity labelling for the first book with
the fourth book and for the second book with the fifth book as well. By the results, it is
reasonable to find further formulations of the labelling for general cases. This work was supported by Doctoral Research Grant, Kementerian Riset dan Teknology, Indonesian
Government No 3117/UN1 DITLIT/DIT LIT/PT/2020 This work was supported by Doctoral Research Grant, Kementerian Riset dan Teknology, Indones This work was supported by Doctoral Research Grant, Kementerian Riset dan Teknology, Indonesian
Government, No.3117/UN1.DITLIT/DIT-LIT/PT/2020. 3. For 𝒎≡𝟐 𝒎𝒐𝒅 𝟑. Under the labelling 𝑓, we have the weights of the edges of the fifth book graph as below: Under the labelling 𝑓, we have the weights of the edges of the fifth book graph as below: 𝜔𝑓 (𝑢5𝑥𝑖,1
5 ) = {
3𝑘4 + 𝑖−1,
3𝑘4 + 𝑖,
3𝑘4 + 𝑖−2,
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
3𝑘4 + 𝑗𝑛+ 𝑖−1,
3𝑘4 + 𝑗𝑛+ 𝑖,
3𝑘4 + 𝑗𝑛+ 𝑖−2,
𝜔𝑓(𝑢5, 𝑣5) =
{
2𝑘4 + 𝑘5 + 𝑛−(
𝑛+1
3 ) ,
2𝑘4 + 𝑘5 + 𝑛−(
𝑛
3) ,
2𝑘4 + 𝑘5 + 𝑛−(
𝑛+2
3 ) ,
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
3𝑘4 + 𝑗𝑛+ 𝑖,
3𝑘4 + 𝑗𝑛+ 𝑖+ 1,
3𝑘4 + 𝑗𝑛+ 𝑖−1,
𝜔𝑓(𝑥𝑖,𝑗
5 𝑥𝑖,𝑗+1
5
) =
{
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛−1
3
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛
3
,
2𝑘4 + 𝑘5 + 𝑖+
(2𝑚−8)𝑛−2
3
𝜔𝑓(𝑥𝑖,𝑗
5 𝑣5) =
{
𝑘4 + 2𝑘5 + 𝑖+
(𝑚−4)𝑛−2
3
,
𝑘4 + 2𝑘5 + 𝑖+
(𝑚−4)𝑛
3
,
𝑘4 + 2𝑘5 + 𝑖+
(𝑚−4)𝑛−4
3
,
with 1 ≤𝑖≤𝑛. 𝑛= 2 (𝑚𝑜𝑑 3)
𝑛= 0 (𝑚𝑜𝑑 3)
𝑛= 1 (𝑚𝑜𝑑 3) 1 ≤𝑗≤
𝑚−2
3 , 𝑛= 2 (𝑚𝑜𝑑 3)
1 ≤𝑗≤
𝑚−2
3 , 𝑛= 0 (𝑚𝑜𝑑 3)
1 ≤𝑗≤
𝑚−2
3 , 𝑛= 1 (𝑚𝑜𝑑 3)
𝑛= 2 (𝑚𝑜𝑑 3)
𝑛= 0 (𝑚𝑜𝑑 3)
𝑛= 1 (𝑚𝑜𝑑 3) 𝑛
1 (𝑚𝑜𝑑 3)
𝑚+1
3
≤𝑗≤𝑚−4, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑚+1
3
≤𝑗≤𝑚−4, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑚+1
3
≤𝑗≤𝑚−4, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3)
𝑗= 𝑚−3, 𝑛= 2 (𝑚𝑜𝑑 3)
𝑗= 𝑚−3, 𝑛= 0 (𝑚𝑜𝑑 3)
𝑗= 𝑚−3, 𝑛= 1 (𝑚𝑜𝑑 3), {
with 1 ≤𝑖≤𝑛. Under the labelling 𝑓, the weights of edges of 5𝐵𝑛(𝐶𝑚) for cases 1, 2 and 3 constitute the
set {3, … , 5(𝑚−1)𝑛+ 7}. This shows that the weights of all the edges of (5𝐵𝑛(𝐶𝑚) are { ,
, (
)
}
different and tes (5𝐵𝑛(𝐶𝑚)) = ⌈
5(𝑚−1)𝑛+7
3
⌉. ◼ {
(
)
}
different and tes (5𝐵𝑛(𝐶𝑚)) = ⌈
5(𝑚−1)𝑛+7
3
⌉. ◼ 9 https://doi.org/10.1051/itmconf/20213603004 ITM Web of Conferences 36, 03004 (2021)
ICMSA 2020 References 1. J.A. Gallian, Electron. J. Comb. (2019) 2. G. Chartrand, M. Jacobson, J. Lehel, O. Oellermann, S. Ruiz, F.Saba, Congr. Numer. 64, 197 – 210 (1988) 3. M. Bača, S. Jendrol, M. Miller, J. Ryan, Discrete Math. 307, 1378–1388 (2007) 4. J. Ivanco, S. Jendrol, Discuss. Math. Graph Theory 26, 449 – 456 (2006) 5. S. Jendrol, J. Miškuf, R. Sotak, Electron. Notes Discret. Math. 28, 281 – 285 (2007) 6. Nurdin, A.N.M. Salman, E.T. Baskoro, J. Comb. Math. Comb. Comput. 65, 163 – 175
(2008) 7. T. Chunling, L. Xiaohui, Y. Yuansheng, W. Liping, Indian J. Pure Appl, Math. 40(3),
155 – 181 (2009) 8. M. Bača, M.K. Siddiqui, Appl Math. Comput. 235, 168 – 173 (2014) 9. A. Ahmad, M. Arshad, G. Izarikova, AKCE Int. J. Graphs Comb. 12, 161 – 168
(2015) 10. M. Bača, S. Jendrol, K. Kathiresan, K. Muthugurupackiam, A.S. Fenovcikova,
Electron. Notes Discret. Math. 48, 19 – 26 (2015) 11. D. Indriati, Widodo, I.E. Wijayanti, K.A. Sugeng, M. Bača, Math. Comput. Sci. 9(2),
161 – 167 (2015) 12. P. Jeyanthi, A. Sudha, Electron. Notes Discret. Math. 48, 175 – 182 (2015) 13. M.K. Siddiqui, D. Afzal, M.R. Faisal, J. Comb. Optim. 34, 534 – 544 (2017) 14. R.W. Putra, Y. Susanti, The total edge irregularity strength of uniform theta graphs, J. Phys. Conf. Ser. 1097, 0012069 (2018) 15. R.W. Putra, Y. Susanti, AKCE Int. J. Graphs Comb. 15, 7 – 13 (2018) 16. L. Ratnasari, S. Wahyuni, Y. Susanti, D.J.E. Palupi, B. Surodjo, Total edge
irregularity strength of arithmetic book graphs, J. Phys. Conf. Ser. 1306, 012032
(2019) 17. L. Ratnasari, S. Wahyuni, Y. Susanti, D.J.E. Palupi, Total edge irregularity strength of
book graphs and double book graphs, AIP Conf. Proc. 2192, 040013 (2019) 18. L. Ratnasari, Y. Susanti, Asian-European J. Math. 13, 2050072 (2020) 19. Y. Susanti, Y.I. Puspitasari, H. Khotimah, Iran. J. Math. Sci. Inform. 15, 1– 13 (2020) 20. L. Ratnasari, S. Wahyuni, Y. Susanti, D.J.E. Palupi, Total edge irregularity strength of
triple book graphs, J. Phys. Conf. Ser. 1772(1), (2020) (Accepted) 10
|
https://openalex.org/W2171749586
|
https://proteomesci.biomedcentral.com/counter/pdf/10.1186/1477-5956-6-27
|
English
| null |
Improved proteome coverage by using iTRAQ labelling and peptide OFFGEL fractionation
|
Proteome science
| 2,008
|
cc-by
| 8,247
|
BioMed Central BioMed Central BioMed Central Open Acc
Research
Improved proteome coverage by using iTRAQ labelling and peptide
OFFGEL fractionation
Emilie Ernoult, Erick Gamelin and Catherine Guette* Email: Emilie Ernoult - emi.ernoult@wanadoo.fr; Erick Gamelin - e.gamelin@unimedia.fr; Catherine Guette* - catherine.guette@univ-angers.fr
* Corresponding author Received: 1 August 2008
Accepted: 13 October 2008 © 2008 Ernoult et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0),
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background: The development of mass spectrometric techniques and fractionation methods now
allows the investigation of very complex protein mixtures ranging from subcellular structures to
tissues. Nevertheless, this work is particularly difficult due to the wide dynamic range of protein
concentration in eukaryotic tissues. In this paper, we present a shotgun method whereby the
peptides are fractionated using OFFGEL electrophoresis after iTRAQ labelling. Results: We demonstrated that iTRAQ peptide labelling enhances MALDI ionisation and that the
OFFGEL fractionation of the labelled peptides introduces a supplementary criterion (pI) useful for
validation and identification of proteins. We showed that iTRAQ samples allowed lower-
concentrated proteins identification in comparison with free-labelled samples. Conclusion: The combined use of iTRAQ labelling and OFFGEL fractionation allows a
considerable increase in proteome coverage of very complex samples prepared from total cell
extracts and supports the low-concentrated protein identification. Proteome Science Open Access Page 1 of 13
(page number not for citation purposes) Background probability of peak overlapping. When iTRAQ-tagged
peptides are subjected to MS/MS analysis, the mass bal-
ancing carbonyl moiety is released as a neutral fragment,
liberating the isotope-encoded reporter ions which pro-
vides relative quantitative information on proteins. g
The iTRAQ-reagent is well known for relative and absolute
quantitation of proteins [1-3]. The interest of this multi-
plexing reagent is that 4 or 8 analysis samples [4] can be
quantified simultaneously. An inherent drawback of the reported iTRAQ technology
is due to the enzymatic digestion of proteins prior to
labelling, which artificially increases sample complexity. Since it has been shown that a reliable determination of
protein dynamics requires quantitative evaluation of an
adequate set of proteolytic peptides derived from each
protein, the iTRAQ approach needs a powerful, multi-
dimensional fractionation method of peptides before MS
identification. In this technique, the introduction of stable isotopes
using iTRAQ reagents occurs on the level of proteolytic
peptides. The iTRAQ technology uses an NHS ester deriv-
ative to modify primary amino groups by linking a mass
balance group (carbonyl group) and a reporter group
(based on N-methylpiperazine) to proteolytic peptides
via the formation of an amide bond. Due to the isobaric
mass design of the iTRAQ reagents, differentially-labelled
peptides appear as a single peak in MS scans, reducing the Page 1 of 13
(page number not for citation purposes) Proteome Science 2008, 6:27 http://www.proteomesci.com/content/6/1/27 Reported peptide separation methods include strong cat-
ion exchange (SCX) chromatography and reverse-phase
chromatography [5]. Recently, isoelectric focusing (IEF), a
high-resolution electrophoresis technique for separation
and concentration of amphoteric biomolecules at their
isoelectric point (pI), has been used in shotgun proteomic
experiments [6]. IEF runs in a buffer-free solution contain-
ing carrier ampholytes or in Immobilized pH gradient
(IPG) gels. Recently, the use of IPG-IEF for the separation
of complex peptide mixtures has been applied to the anal-
ysis of plasma and amniotic fluid [7,8] as well as to bacte-
rial material [9]. However, a major limitation of this
method is the tedious post-IEF sample processing. The
IPG gel strip is divided into small sections for extraction
and cleaning up of the peptides. A new concept called
OFFGEL electrophoresis was recently introduced with the
primary aim of purifying proteins and peptides [10]. This
technique recovers the sample from the liquid phase and
was demonstrated to be of great interest in shotgun pro-
teomics [11]. Background IEF is not only a high resolution and high
capacity separation method for peptides, it also provides
additional physicochemical information like their isoelec-
tric point [12,13]. The pI value provided is used as an
independent validating and filtering tool during database
search for MS/MS peptide sequence identification [14]. wide pI-range (pH 3–10) and compared the proteome
coverage between free-samples and iTRAQ-samples. Results and discussion
The influence of iTRAQ-reagent tagging g
gg
g
The 2D-LC separation of 200 μg of free-labelled digested
proteins by SCX allowed identification of 159 proteins
among which 116 proteins were characterised by at least
2 peptides (73% performance) (Table 1, NL-116). From
four fractions of 50 μg of proteins which were reduced,
blocked with MMTS, digested with trypsin, labelled using
a different iTRAQ reagent, pooled and separated by SCX
chromatography (iTRAQ-310), 472 proteins can be iden-
tified. 310 out of these have been identified using at least
two peptides (Table 1). All the 116 proteins that were
identified without labelling (NL-116) were also identified
in the iTRAQ experiment. Nevertheless, iTRAQ labelling
allowed 2.7 more proteins identified (by at least 2 pep-
tides) compared to free-labelled experiments. All experiments: 1111 proteins, with 2 pept: 947 proteins Increase in peptide mass following iTRAQ labelling We know that tertiary amines have a stronger
proton affinity than the one primary amines (about 50 kJ/
mole). By introducing 2 tertiary amine functions on pep-
tides having lysines, their proton affinity and therefore
their ionisation increase. For example, Figure 2 shows MS/
MS spectra of the peptide GALQNIIPASTGAAK before and
after iTRAQ labelling. The signal/noise ratio is multiplied
by 6 for the most intense y8 ion. This result suggests that
the proton affinity of iTRAQ modified lysines is almost
equal to the one of arginines. We found that iTRAQ gave
equivalent identification success rate for arginine- and
lysine-terminated peptides, hence increasing the peptide
coverage of proteins and increasing the chance of identify-
ing low-concentration peptides. iTRAQ increases ionisation of peptides containing lysines
Analysis of the non-labelled peptide sequences showed
that 20% of them had a lysine termination, and 80% an
arginine termination (Figure 1). A recent study demon-
strated that during MALDI ionisation, there is a linear cor-
relation between peptide ionisation levels and the proton
affinity of the amino acids, with the exception of lysine
[20]. Arginine is the residue having the strongest proton
affinity, and peptides containing this amino acid are those
ionising the best. This conclusion is in line with our
results; peptides containing arginine had better ionisation
levels, therefore providing a better signal to noise ratio for
simple MS, and thus providing higher quality MS/MS
spectra responsible for more valid sequence identification
than peptides having a Cterm lysine. The analysis of peptides identified following iTRAQ label-
ling showed a balance of the number of arginine- or
lysine-terminated peptides with 50% arginine-terminated
and 50% lysine-terminated (Figure 1). Our results con-
firm those of Ross [1] who showed that in comparable
whole-yeast experiments using electrospray, (much less
sensitive to proton affinity than MALDI ionisation), the
ratio of lysine to arginine-terminated peptides identified
increased from 0.79 for native peptide, to 0.98 for iTRAQ The influence of the quantity of iTRAQ-labelled peptides
From only 400 μg of reduced, blocked, digested cellular
lysate labelled using iTRAQ, we were able after SCX sepa-
ration to identify, 492 proteins, of which 429 with at least
2 peptides, giving an identification rate of 87% (iTRAQ-
429). By doubling the amount of biological material to iTRAQ labelling favours the peptide ionization
Figure 1
iTRAQ labelling favours the peptide ionization. Increase in peptide mass following iTRAQ labelling p p
f
g
Q
g
The presence of iTRAQ label on the NH2 terminal adds a
mass of 145 to the peptide m/z. If the peptide had a lysine
residue at the Cterm position, two iTRAQ labels are added
(290 Da). Looking at the list of peptides identified in the
iTRAQ-310 label experiment, 61 unique peptides without
lysine in their sequence had a m/z between 800 and 945,
and 55 other peptides with a lysine at the Cterm extremity
had a mass between 800 and 1090. These 116 peptides
were not visible without iTRAQ labelling; nevertheless,
they were found in the sequence of 60 proteins already
identified by at least 2 peptides. Only 3 proteins were
identified by a second peptide. The mass increase due to
iTRAQ-labelling cannot be the unique explanation of the
higher number of proteins identified nor the higher
number of peptides contributing to the identification for
each protein. Recently, the compatibility of iTRAQ isotope labelling
and OFFGEL-IEF for relative quantification and validation
of sequence matches from database searching was shown
from a BSA tryptic digest sample and complex eukaryotic
samples [15,16] but surprisingly, no attempts was done to
undertake comprehensive analysis of influence of iTRAQ
labelling on the proteome coverage ratio. In our work, we combined free-labelled peptides or
iTRAQ labelled peptides and OFFGEL fractionation for
the proteomic study of a very complex sample like the
human neuroblastoma SH-SY5Y cell line [17-19] in a Page 2 of 13
(page number not for citation purposes)
Table 1: The list of experiments and the number of identified proteins
Experiment
Quantity (μg)
Fractionation
Labelling
Total proteins
Proteins with 2 peptides (at least)
Yield (%)
NL-116
200
SCX
no
159
116
73
NL-184
200
OFFGEL-IEF
no
285
184
64
NL-235
200
SCX + OFFGEL-IEF
no
302
235
78
iTRAQ-310
200
SCX
iTRAQ
472
310
66
iTRAQ-429
400
SCX
iTRAQ
492
429
87
iTRAQ-739
400
OFFGEL-IEF
iTRAQ
879
739
84
All experiments: 1111 proteins, with 2 pept: 947 proteins Table 1: The list of experiments and the number of identified proteins http://www.proteomesci.com/content/6/1/27 http://www.proteomesci.com/content/6/1/27 Proteome Science 2008, 6:27 derived peptides. The reporter group of the iTRAQ reagent
is a piperazine group having 2 tertiary amines groups. By
attaching these onto the lysine's lateral chain, a primary
amine function is thereby replaced by two tertiary amine
functions. Increase in peptide mass following iTRAQ labelling Number of identified peptides with a lysine or an arginine in the C-term
position; white bars: free-labelled peptides; black bars: iTRAQ peptides. 900
800
700
number of peptides
600
500
iTRAQ peptides
free-labelled peptides
400
300
200
100
0
K
R
C-term amino acid iTRAQ labelling favours the peptide ionization
Figure 1
iTRAQ labelling favours the peptide ionization. Number of identified peptides with a lysine or an arginine in the C-term
position; white bars: free-labelled peptides; black bars: iTRAQ peptides. iTRAQ labelling favours the peptide ionization
Figure 1
iTRAQ labelling favours the peptide ionization. Number of identified peptides with a lysine or an arginine in the C-term
position; white bars: free-labelled peptides; black bars: iTRAQ peptides. Page 3 of 13
(page number not for citation purposes) http://www.proteomesci.com/content/6/1/27 Proteome Science 2008, 6:27 MS/MS spectra of GALQNIIPASTGAAK (which allowed the identification of the Glyceraldehyde 3 phosphate deshydrogenase
rom 200 μg of digested proteins fractionated by SCX chromatography
igure 2
MS/MS spectra of GALQNIIPASTGAAK (which allowed the identification of the Glyceraldehyde 3 phosphate
eshydrogenase) from 200 μg of digested proteins fractionated by SCX chromatography. (A) free-labelled expe
ment; (B) iTRAQ labelling experiment. start with, identification rate for proteins with at least 2
peptides raised by 39% (Table 1). largely due to the Bjellqvist algorithm [12] which is used
in this study. This algorithm is known to overestimate pI
values in the acidic range [22]. One feature of this experi-
ment is that after screening with our previously-described
criteria, no peptide was identified in fraction 24 (Figure
4B); this is in line with Fraterman's work [23]. In order to
verify the identification of the peptides set and decrease
false positive percentage after ProteinPilot calculations,
we used the pI peptide property as an orthogonal property
of these peptides of interest [24]. By separating peptides
using OFFGEL, we showed that we can predict their pI
with an accuracy of 0.4 pI unit. Under this condition, all
peptides with unused ProteinPilot score > 1.3 and show-
ing an experimental pI deviating by more than 0.4 pI unit
from calculated value were excluded. All free-labelled pep-
tides presented in this work were filtered according to
their ProteinPilot unused score and pI as described above. The influence of the fractioning method on free-labelled
peptides p p
The OFFGEL technology of 200 μg of free-labelled
digested proteins allowed identification of 285 proteins,
including 184 with at least 2 peptides (NL-184). By look-
ing at the number of fractions in which each distinct pep-
tide is found we can judge the fractionation quality of the
technique. Figure 3 shows that 84% of the identified pep-
tides are found in only one fraction and more than 95%
are found in one or two fractions. This result is in agree-
ment with the results found from earlier studies [21]. The
average experimental pH value of each fraction is pre-
sented as a bar in Figure 4A. The theoretical pH values pro-
vided by the manufacturer were overlaid as a broken line. Average experimental pH values deviated from theoretical
values by an average error of 0.29 pI unit. Only fractions
close to neutrality (Fractions F12 to F18), and which do
not contain a large number of peptides, showed an error
of 0.38 pI unit in agreement with already published
results [22]. Such an error was also found for the 3 more
acidic fractions F1 to F3. For these 3 fractions, the error is Increase in peptide mass following iTRAQ labelling
M
/z)
40
50
60
70
80
90
100
ass (m
10
20
30
9.0
374.8
740.6
1106.4
1472.2
209.6
0
Prec = 1411.79
y14
1354.76(S92)
y8
702.37(S47)
129.06(S38)
b4
370.20(S38)
b5
484.25(S21)
86.111(S18)
y10
928.54(S18)
y11
1042.58(S10)
y9
815.614(S11)
A
9.0
366.4
723.8
1081.2
1438.6
1796.0
Mass (m/z)
266.0
0
10
20
30
40
50
60
70
80
90
100
Prec = 1699.91
114.11(S308)
y8
846.48(S288)
b7
854.52(S120)
b5
628.35(S85)b6
741.43(S62)y9
959.56(S47)
70.06(S37)
b3
386.25(S28)
86.09(S19)
y6
y11
1186.69(S15)
B A
M
/z)
40
50
60
70
80
90
100
ass (m
10
20
30
9.0
374.8
740.6
1106.4
1472.2
209.6
0
Prec = 1411.79
y14
1354.76(S92)
y8
702.37(S47)
129.06(S38)
b4
370.20(S38)
b5
484.25(S21)
86.111(S18)
y10
928.54(S18)
y11
1042.58(S10)
y9
815.614(S11)
A Prec = 1411.79 y11
1042.58(S10) 9.0
366.4
723.8
1081.2
1438.6
1796.0
Mass (m/z)
266.0
0
10
20
30
40
50
60
70
80
90
100
Prec = 1699.91
114.11(S308)
y8
846.48(S288)
b7
854.52(S120)
b5
628.35(S85)b6
741.43(S62)y9
959.56(S47)
70.06(S37)
b3
386.25(S28)
86.09(S19)
y6
y11
1186.69(S15)
B B MS/MS spectra of GALQNIIPASTGAAK (which allowed the identification of the Glyceraldehyde 3 phosphate deshydrogenase)
from 200 μg of digested proteins fractionated by SCX chromatography
Figure 2
MS/MS spectra of GALQNIIPASTGAAK (which allowed the identification of the Glyceraldehyde 3 phosphate
deshydrogenase) from 200 μg of digested proteins fractionated by SCX chromatography. (A) free-labelled exper-
iment; (B) iTRAQ labelling experiment. 9.0
366.4
723.8
1081.2
1438.6
1796.0
Mass (m/z)
266.0
0
10
20
30
40
50
60
70
80
90
100
Prec = 1699.91
114.11(S308)
y8
846.48(S288)
b7
854.52(S120)
b5
628.35(S85)b6
741.43(S62)y9
959.56(S47)
70.06(S37)
b3
386.25(S28)
86.09(S19)
y6
y11
1186.69(S15)
B MS/MS spectra of GALQNIIPASTGAAK (which allowed the identification of the Glyceraldehyde 3 phosphate deshydrogenase)
from 200 μg of digested proteins fractionated by SCX chromatography
Figure 2
MS/MS spectra of GALQNIIPASTGAAK (which allowed the identification of the Glyceraldehyde 3 phosphate
deshydrogenase) from 200 μg of digested proteins fractionated by SCX chromatography. (A) free-labelled exper-
iment; (B) iTRAQ labelling experiment. Page 4 of 13
(page number not for citation purposes) Proteome Science 2008, 6:27 http://www.proteomesci.com/content/6/1/27 start with, identification rate for proteins with at least 2
peptides raised by 39% (Table 1). The quality of OFFGEL separation of iTRAQ-labelled
peptides By using OFFGEL separation by 24 fractions of 400 μg of
iTRAQ-labelled peptides, we identified 879 proteins, of
which 739 with at least 2 peptides (iTRAQ-739), giving an
increase of 72% (Table 1) compared to SCX separation. Fractionwise distribution of identified SH-SY5Y peptides
Figure 3
Fractionwise distribution of identified SH-SY5Y peptides. white bars: free-labelled peptides; black bars: iTRAQ pep-
tides. 0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
1
2
3
4
Fraction numbers containing each peptide
Total identified peptides (%)
free-labelled peptides
iTRAQ peptides free-labelled peptides
iTRAQ peptides free-labelled peptides
iTRAQ peptides 2 Fraction numbers containing each peptide Fractionwise distribution of identified SH-SY5Y peptides
Figure 3
Fractionwise distribution of identified SH-SY5Y peptides. white bars: free-labelled peptides; black bars: iTRAQ pep-
tides. Fractionwise distribution of identified SH-SY5Y peptides
Figure 3
Fractionwise distribution of identified SH-SY5Y peptides. white bars: free-labelled peptides; black bars: iTRAQ pep-
tides. identified SH SY5Y peptides
n of identified SH-SY5Y peptides. white bars: free-labelled peptides; black bars: iTRAQ pep- Page 5 of 13
(page number not for citation purposes) oteome Science 2008, 6:27
http://www.proteomesci.com/content/6
nalysis of SH-SY5Y peptides pI after OFFGEL fractionation and MALDI- MS/MS identification
gure 4
nalysis of SH-SY5Y peptides pI after OFFGEL fractionation and MALDI- MS/MS identification. (A) The ave
perimental pH of all peptides in a single fraction after filtering for false positive is presented as bars; white bars: non-lab
ptides; dark bars: iTRAQ peptides. Error bars indicate the SD of each fraction's experimental pI. The broken line is base
e theoretical pI values for an IPG strip of 24 cm ranging from pH 3–10; Agilent Technologies provided the theoretical p
s. (B) The total bar height shows the number of peptides in each fraction after filtering for false positive; white bars: no
belled peptides; black bars: iTRAQ peptides. The quality of OFFGEL separation of iTRAQ-labelled
peptides 3
4
5
6
7
8
9
10
11
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
Fraction number
Isoelectric point
non-labelled peptides
iTRAQ peptides
A
0
100
200
300
400
500
600
700
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
Fraction number
Number of identified peptides
free-labelled peptides
iTRAQ peptides
B http://www.proteomesci.com/content/6/1/27 Proteome Science 2008, 6:27 3
4
5
6
7
8
9
10
11
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
Fraction number
Isoelectric point
non-labelled peptides
iTRAQ peptides
A Analysis of SH-SY5Y peptides pI after OFFGEL fractionation and MALDI- MS/MS identification
Figure 4
Analysis of SH-SY5Y peptides pI after OFFGEL fractionation and MALDI- MS/MS identification. (A) The average
experimental pH of all peptides in a single fraction after filtering for false positive is presented as bars; white bars: non-labelled
peptides; dark bars: iTRAQ peptides. Error bars indicate the SD of each fraction's experimental pI. The broken line is based on
the theoretical pI values for an IPG strip of 24 cm ranging from pH 3–10; Agilent Technologies provided the theoretical pI val-
ues. (B) The total bar height shows the number of peptides in each fraction after filtering for false positive; white bars: non-
labelled peptides; black bars: iTRAQ peptides. 0
100
200
300
400
500
600
700
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
Fraction number
Number of identified peptides
free-labelled peptides
iTRAQ peptides
B 0
100
200
300
400
500
600
700
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
Fraction number
Number of identified peptides
free-labelled peptides
iTRAQ peptides
B Analysis of
Figure 4 Analysis of
Figure 4 Analysis of SH-SY5Y peptides pI after OFFGEL fractionation and MALDI- MS/MS identification
Figure 4
Analysis of SH-SY5Y peptides pI after OFFGEL fractionation and MALDI- MS/MS identification. (A) The average
experimental pH of all peptides in a single fraction after filtering for false positive is presented as bars; white bars: non-labelled
peptides; dark bars: iTRAQ peptides. Error bars indicate the SD of each fraction's experimental pI. The broken line is based on
the theoretical pI values for an IPG strip of 24 cm ranging from pH 3–10; Agilent Technologies provided the theoretical pI val-
ues. (B) The total bar height shows the number of peptides in each fraction after filtering for false positive; white bars: non-
labelled peptides; black bars: iTRAQ peptides. Page 6 of 13
(page number not for citation purposes) http://www.proteomesci.com/content/6/1/27 Proteome Science 2008, 6:27 OFFGEL experiment also increased the number of pep-
tides identified per protein from 4.8 (iTRAQ-429) to 7.1
(iTRAQ-739), thereby increasing dramatically the confi-
dence level of the protein identification. Figure 5 shows
the quantity of proteins identified with different numbers
of matching peptides with the two types of fractionation. We can see that OFFGEL gave a higher number of proteins
identified in all fractions. imental pI value deviated from the theoretical pI value by
an average error of +/- 0.43. In the pH range of 3 to 8, the
average pI value of the labelled peptides fits very well with
the average value of the non-labelled peptides. Major
deviations were observed at the basic pH range (8.3–10.0)
for fractions 18–24. By splitting OFFGEL fractionation
into 2 zones, we noticed that the mean error in the pH 3–
8 range was +/- 0.34 and was rising to +/- 0.64 in the more
alkaline range (pH 8–10); this can probably be explained
by the fact that the software used for calculating the pI
value does not take into account the presence of the
iTRAQ label. As for the free-labelled peptides, we intro-
duced the pI iTRAQ-labelled peptides property as a filter
to verify the identification of the set of peptides. For the
fractions F1 to F17 (pH 3.3–8.0 range), all peptides with
an unused ProteinPilot score above 1.3 and with an exper-
imental pI difference larger than 0.4 pI unit were
excluded. For the fractions F18-F24 (pH 8.3–10.0), we
excluded all peptides with experimental pI difference
greater than 0.7 pI unit. Analysis of
Figure 4 All iTRAQ-labelled peptides pre-
sented in this work were filtered using this protocol. As shown in Figure 3, 78% of the identified peptides are
found in only one fraction and about 90% are found in
one or two fractions, confirming the distribution of free-
labelled peptides in our work and the results from earlier
studies [16,21]. The distribution of peptides per fraction is shown in Fig-
ure 4. The pI value for each identified peptide was calcu-
lated by using Bjellqvist's algorithm without taking into
account the iTRAQ groups in N-term position and/or on
the lateral lysine chain. Using these data, average pI values
with standard deviations were calculated for all peptides
identified in each fraction (Figure 4A). The average exper- The distribution of identified proteins based on the number of peptides used for protein identification
Figure 5
The distribution of identified proteins based on the number of peptides used for protein identification Compar
0
50
100
150
200
250
2
3
4
5
6
7
8
9
>10
Peptide Matching Number
Number of identified proteins
iTRAQ-429 (SCX)
iTRAQ-739 (OFFGEL) The distribution of identified proteins based on the number of peptides used for protein identification
Figure 5
The distribution of identified proteins based on the number of peptides used for protein identification. Compar-
ison between SCX and OFFGEL fractionation of iTRAQ samples. 0
50
100
150
200
250
2
3
4
5
6
7
8
9
>10
Peptide Matching Number
Number of identified proteins
iTRAQ-429 (SCX)
iTRAQ-739 (OFFGEL) iTRAQ-429 (SCX)
iTRAQ-739 (OFFGEL) iTRAQ-429 (SCX)
iTRAQ-739 (OFFGEL) Page 7 of 13
(page number not for citation purposes)
The distribution of identified proteins based on the number of peptides used for protein identification
Figure 5
The distribution of identified proteins based on the number of peptides used for protein identification. Compar-
ison between SCX and OFFGEL fractionation of iTRAQ samples. The distribution of identified proteins based on the number of peptides used for protein identification
Figure 5
The distribution of identified proteins based on the number of peptides used for protein identification. Compar-
ison between SCX and OFFGEL fractionation of iTRAQ samples. Page 7 of 13
(page number not for citation purposes) Proteome Science 2008, 6:27 http://www.proteomesci.com/content/6/1/27 Proteomic coverage fractionation and better membrane-solubilising strate-
gies. The molecular weight (MW) distribution of the iden-
tified proteins is shown in Figure 7. The overall trend of
the MW distribution of the identified proteins versus the
human proteome is very similar, even if proteins in the
<20 kDa range is slightly higher suggesting there is no real
bias in our protocol. Combining results of all experiments, we end up with 947
proteins (Additional file1) identified with at least 2 pep-
tides and 6119 unique peptides. 380 detected proteins
have at least 5 or more unique peptides identified. The proteins observed in our experiments were mapped in
relation with their MW and pI. The distribution of pI val-
ues over a range of 3–12 is shown in Figure 6. Similar to
previous computational analyses [25,26] our results show
that the distribution of the predicted pI values of our iden-
tified proteins is typical of eukaryotic cells, showing a kind
of tri-modal distribution. The pI value distribution of our
identified proteins shows the same profile as the distribu-
tion of human proteins identified from epithelial mam-
mary cells (quite different from human proteome) [25]. Our results show a fairly good coverage for both cytosolic
and membrane proteins. Indeed, the coverage is slightly
biased towards the identification of cytosolic proteins
(cluster at pI 6), presumably due to a better solubility of
cytosolic proteins compared to membrane proteins (clus-
ter at pI 9). Improved coverage of membrane proteins
from plasma membrane and subcellular organelles can be
achieved by coupling OFFGEL with further subcellular By comparing MW profiles of the proteins identified in
the iTRAQ-947 and iTRAQ-739 experiments, we can see
that the profiles are comparable (Figure 7), strengthening
the hypothesis that the iTRAQ-739 experiment correctly
reflects proteome coverage of the cell line SH-SY5Y. Relative abundance of proteins One way to get an idea of the quantity of a protein in a
complex mixture is to calculate its PAI [27], which repre-
sents the number of identified peptides divided by the
number of theoretical tryptic peptides. The PAI mean
value for the iTRAQ-947 experiment is 0.32; 50% of the
proteins have a PAI value < 0.14 and 21% have a PAI value
< 0.05, a characteristically value for proteins present at
low concentration. Some of these low abundance proteins
such as Transcription factor BTF3 homologue 4, Tran- Protein distribution in function of pI
Figure 6
Protein distribution in function of pI. The isoelectric point distribution of SH-SY5Y proteins identified with 2 or more
peptides. 0
5
10
15
20
25
30
35
3.7-3.8
4-4.1
4.3-4.4
4.6-4.7
4.9-5
5.2-5.3
5.5-5.6
5.8-5.9
6.1-6.2
6.4-6.5
6.7-6.8
7-7.1
7.3-7.4
7.6-7.7
7.9-8
8.2-8.3
8.5-8.6
8.8-8.9
9.1-9.2
9.4-9.5
9.7-9.8
10-10.1
10.3-10.4
10.6-10.7
10.9-11
11.2-11.3
11.5-11.6
11.8-11.9
12.1-12.2
Isoelectric point
Number of proteins Protein distribution in function of pI
Figure 6
Protein distribution in function of pI. The isoelectric point distribution of SH-SY5Y proteins identified with 2 or more
peptides. Protein distribution in function of pI
Figure 6
Protein distribution in function of pI. The isoelectric point distribution of SH-SY5Y proteins identified with 2 or more
peptides. n function of pI
n in function of pI. The isoelectric point distribution of SH-SY5Y proteins identified with 2 or more Page 8 of 13
(page number not for citation purposes) Proteome Science 2008, 6:27
http://www.proteomesci.com/content/6/1/27
Protein distribution in function of MW
Figure 7
Protein distribution in function of MW. The molecular weight distribution of SH-SY5Y proteins identified with two or
more peptides; white bars: from one experiement with 400 μg of digested proteins and labelled with iTRAQ reagent ; black
bars: from all the experiments
0
50
100
150
200
250
<20
20-40
40-60
60-80
80-100
100-120
120-140
140-160
160-180
180-200
200-220
220-240
240-260
260-280
280-300
>300
Molecular weight
Number of proteins
iTRAQ-739
iTRAQ-947 http://www.proteomesci.com/content/6/1/27 Proteome Science 2008, 6:27 Molecular weight Protein distribution in function of MW
Figure 7
Protein distribution in function of MW. Relative abundance of proteins The molecular weight distribution of SH-SY5Y proteins identified with two or
more peptides; white bars: from one experiement with 400 μg of digested proteins and labelled with iTRAQ reagent ; black
bars: from all the experiments clearly see the superiority of iTRAQ labelling in compari-
son to non-labelled molecules, and the superiority of the
OFFGEL-IEF fractionation method as compared to the
SCX-C18/2D-LC method. scription initiation factor IIF sub-unit beta, and Nuclear
factor 1 B-type are identified in this range of low PAI lev-
els. The comparison between an experiment without
labelling and an experiment with iTRAQ labelling, using
similar method of separation, shows a higher number of
proteins identified in each category for iTRAQ-labelled
proteins (Figure 8A). This result is particularly convincing
when looking at very low PAI values, and hence at pro-
teins present at low concentration. In the area of PAI levels
lower than 0.05, only 2 proteins were identified in the
non-labelling experiment, compared to 44 identified fol-
lowing iTRAQ labelling. We carried out comparisons for
similar PAI values for SCX-C18/2D-LC and OFFGEL-IEF
fractionation. Starting with only 400 μg of proteins with
iTRAQ labelling, we demonstrated that OFFGEL-IEF sepa-
ration gives the largest number of identified proteins, par-
ticularly in areas with low PAI values (Figure 8B). By
comparing the PAI values of proteins identified with at
least 2 peptides in the different experiments, we can scription initiation factor IIF sub-unit beta, and Nuclear
factor 1 B-type are identified in this range of low PAI lev-
els. The comparison between an experiment without
labelling and an experiment with iTRAQ labelling, using
similar method of separation, shows a higher number of
proteins identified in each category for iTRAQ-labelled
proteins (Figure 8A). This result is particularly convincing
when looking at very low PAI values, and hence at pro-
teins present at low concentration. In the area of PAI levels
lower than 0.05, only 2 proteins were identified in the
non-labelling experiment, compared to 44 identified fol-
lowing iTRAQ labelling. We carried out comparisons for
similar PAI values for SCX-C18/2D-LC and OFFGEL-IEF
fractionation. Starting with only 400 μg of proteins with
iTRAQ labelling, we demonstrated that OFFGEL-IEF sepa-
ration gives the largest number of identified proteins, par-
ticularly in areas with low PAI values (Figure 8B). By
comparing the PAI values of proteins identified with at
least 2 peptides in the different experiments, we can Page 9 of 13
(page number not for citation purposes) Conclusion
i
i iTRAQ is at evidence a very powerful tool, recognised for
its ability to relatively quantify proteins. In this work, we
showed that the iTRAQ reagent improves MALDI ionisa-
tion, especially for peptides containing lysine. A direct
consequence of this property is the better chance of iden-
tifying low abundance proteins in complex biological
materials. We were also able to demonstrate that an OFF-
GEL fractionation step, has a positive influence on the
number of proteins identified compared to SCX method. From standard clinical protein quantities (400 μg), we
proposed a methodology allowing the identification of
more than 800 proteins. Having established a protein Page 9 of 13
(page number not for citation purposes) Page 9 of 13
(page number not for citation purposes) Proteome Science 2008, 6:27
http://www.proteomesci.com/content/6
Protein abundance index (PAI) for the identified proteins
Figure 8
Protein abundance index (PAI) for the identified proteins. (A) influence of the iTRAQ labelling: free-labelled vs iT
peptides (from 200 μg of digested proteins fractionated by SCX chromatography); (B) influence of the fractionation mod
SCX vs OFFGEL (from 400 μg of digested proteins and labelled with iTRAQ reagent). 0
10
20
30
40
50
60
<0.05
0.05-0.1
0.1-0.2
0.2-0.3
0.3-0.4
0.4-0.5
0.5-0.6
0.6-0.7
0.7-0.8
0.8-0.9
0.9-1
>1
PAI
Free-labelled peptides
iTRAQ peptides
A
Number of identified proteins
0
20
40
60
80
100
120
140
160
180
<0.05
0.05-0.1
0.1-0.2
0.2-0.3
0.3-0.4
0.4-0.5
0.5-0.6
0.6-0.7
0.7-0.8
0.8-0.9
0.9-1.0
>1
PAI
Number of identified proteins
SCX
OFFGEL
B http://www.proteomesci.com/content/6/1/27 Proteome Science 2008, 6:27 0
10
20
30
40
50
60
<0.05
0.05-0.1
0.1-0.2
0.2-0.3
0.3-0.4
0.4-0.5
0.5-0.6
0.6-0.7
0.7-0.8
0.8-0.9
0.9-1
>1
PAI
Free-labelled peptides
iTRAQ peptides
A
Number of identified proteins A 0
20
40
60
80
100
120
140
160
180
<0.05
0.05-0.1
0.1-0.2
0.2-0.3
0.3-0.4
0.4-0.5
0.5-0.6
0.6-0.7
0.7-0.8
0.8-0.9
0.9-1.0
>1
PAI
Number of identified proteins
SCX
OFFGEL
B Protein abundance index (PAI) for the identified proteins
Figure 8
Protein abundance index (PAI) for the identified proteins. (A) influence of the iTRAQ labelling: free-labelled vs iTRAQ
peptides (from 200 μg of digested proteins fractionated by SCX chromatography); (B) influence of the fractionation mode:
SCX vs OFFGEL (from 400 μg of digested proteins and labelled with iTRAQ reagent). Protein digestion and peptide labelling with iTRAQ
reagents g
Protein samples were cleaned up by precipitation with 6
volumes of cold acetone at -20°C overnight followed by
resuspension of pellets in 0.5M triethylammonium bicar-
bonate (TEAB) pH 8.5 (Sigma-Aldrich) and final centrifu-
gation step at 14,000 × g at 4°C for 15 min. We quantified
proteins from supernatant with the 2-D Quant Kit (GE
Healthcare, München, Germany) before diluting the pro-
tein samples up to 5 mg/ml with TEAB buffer. We took 50
or 100 μg of proteins for further reduction, alkylation,
digestion and iTRAQ labelling using iTRAQ Reagents Mul-
tiplex Kit (Applied Biosystems) according to manufac-
turer's protocol. Briefly, protein samples were reduced
with 5 mM tris-(2-carboxyethyl)phosphine (TCEP) at
60°C for 1 h. and the cysteine-groups were blocked using
a 10 mM methyl methanethiosulfonate (MMTS) solution
at room temperature for 10 min. . The proteins were then
digested by 10 μg of trypsin at 37°C for 16 h. Each peptide
solution was labelled at room temperature for 1 h with
one iTRAQ reagent vial (mass tag 114, 115, 116 or 117)
previously reconstituted with 70 μl of ethanol. Samples of
the same protein content, and labelled respectively with
114, 115, 116 and 117 iTRAQ reagents, were combined
and labelling reaction stopped by evaporation in a Speed
Vac to obtain a brown pellet. Peptides were desalted for 3 min. using only buffer A on
the precolumn, followed by a separation for 60 min. using
the following gradient: 0 to 20% B in 10 min., 20% to
55% B in 45 min. and 55% to 100% B in 5 min. Chroma-
tograms were recorded at the wavelength of 214 nm. Pep-
tide fractions were collected using a Probot microfraction
collector (Dionex). For the SCX fractionation, we used a salt gradient steps:
20-μl injections of 5 mM, 10 mM, 25 nM, 50 mM, 75
mM,100 mM, 125 mM, 150 mM, 200 mM, 300 mM, 500
mM, 1000 mM NaCl For the SCX fractionation, we used a salt gradient steps:
20-μl injections of 5 mM, 10 mM, 25 nM, 50 mM, 75
mM,100 mM, 125 mM, 150 mM, 200 mM, 300 mM, 500
mM, 1000 mM NaCl We used CHCA (LaserBioLabs, Sophia-Antipolis, France)
as MALDI matrix. The matrix (concentration of 2 mg/mL
in 70% ACN in water with 0.1% TFA) was continuously
added to the column effluent via a micro "T" mixing piece
at 1.2 μL/min flow rate. Capillary LC separation The samples were separated on an Ultimate 3,000 nano-
LC system (Dionex, Sunnyvale, USA) using a C18 column
(PepMap100, 3 μm, 100A, 75 μm id × 15 cm, Dionex) at
a flow rate of 300 nL/min. . Buffer A was 2% ACN in water
with 0.05% TFA and buffer B was 80% ACN in water with
0.04% TFA. Protein digestion and peptide labelling with iTRAQ
reagents After 14 min run, a start signal
was sent to the Probot to initiate fractionation. Fractions
were collected for 10s and spotted on a MALDI sample
plate (1,664 spots per plate, Applied Biosystems, Foster
City, CA). Materials and methods
Cell culture and protein extraction The human neuroblastoma cell line SH-SY5Y was a gift
from Dr F. Vallette (INSERM U601, Nantes, France). Cells
were grown in RPMI-1640 medium (Lonza) supple-
mented with 10% (v/v) fetal bovine serum (Lonza) with-
out antibiotics in a, 5% humid CO2 atmosphere at 37°C. Cells were scraped and washed 3 times with PBS (300 × g,
5 min.). Cell pellets were then lysed by a solution contain-
ing 7M urea, 2M thiourea, 4% (w/v) CHAPS at 4°C for 1
h using a rotary shaker. Lysis was achieved by sonication
on ice (3 × 5s pulses), and the lysates were clarified by cen-
trifugation at 14,000 × g at 4°C for 15 min. Conclusion
i
i 0
20
40
60
80
100
120
140
160
180
<0.05
0.05-0.1
0.1-0.2
0.2-0.3
0.3-0.4
0.4-0.5
0.5-0.6
0.6-0.7
0.7-0.8
0.8-0.9
0.9-1.0
>1
PAI
Number of identified proteins
SCX
OFFGEL
B B Protein abundance index (PAI) for the identified proteins
Figure 8
Protein abundance index (PAI) for the identified proteins. (A) influence of the iTRAQ labelling: free-labelled vs iTRAQ
peptides (from 200 μg of digested proteins fractionated by SCX chromatography); (B) influence of the fractionation mode:
SCX vs OFFGEL (from 400 μg of digested proteins and labelled with iTRAQ reagent). http://www.proteomesci.com/content/6/1/27 http://www.proteomesci.com/content/6/1/27 Proteome Science 2008, 6:27 database of SH-SY5Y cells, we can now use it as a reference
for further research in this domain. the manufacturer for 15 min. Then, 150 μL of sample was
loaded in each well. Electrofocusing of the peptides is per-
formed at 20°C and 50 μA until the 50 kVh level was
reached. After focusing, the 24 peptide fractions were
withdrawn and the wells rinsed with 200 μL of a solution
of water/methanol/formic acid (49/50/1) after 15 min,
the rinsing solutions were pooled with their correspond-
ing peptide fraction. All fractions were evaporated by cen-
trifugation under vacuum and maintained at -20°C. Just
prior nano-LC, the fractions were resuspended in 20 μL of
H2O with 0.1% (v/v) TFA. Page 11 of 13
(page number not for citation purposes) References 1. Ross PL, Huang YN, Marchese JN, Williamson B, Parker K, Hattan S,
Khainovski N, Pillai S, Dey S, Daniels S, et al.: Multiplex protein
quantitation in Saccharomyces cerevisiae using amine-reac-
tive isobaric tagging reagent. Mol Cell Proteomics 2004,
3:1154-1169. While this software automatically accepts all peptides hav-
ing an identification confidence level >1%, only proteins
having at least one peptide above 95% confidence were
initially recorded. The low confidence peptides cannot
give a positive protein identification by themselves, but
may support the presence of a protein identified using
other peptides with higher confidence. Performing the
search against a concatenated database containing both
forward and reversed sequences allowed estimation of the
false discovery rate below1%. 2. Chong PK, Gan CS, Pham TK, Wright PC: Isobaric tags for rela-
tive and absolute quantification (iTRAQ) reproductibility:
implication of multiple injections. J Proteome Res 2006,
5:1232-1240. 3. Gan CS, Chong PK, Pham TK, Wright PC: Technical, experimen-
tal, and biological variations in isobaric tags for relative and
absolute quantitation (iTRAQ). J Proteome Res 2007, 6:821-827. 4. Pierce A, Unwin RD, Evans CA, Griffiths S, Carney L, Zhang L, Jawor-
ska E, Lee CF, Blinco D, Okoniewski MJ, et al.: Eight-channel
iTRAQ enables comparison of the activity of 6 leu-
kaemogenic tyrosine kinases. Mol Cell Proteomics 2007, 7:853-83. 5. Wolters DA, Washburn MP, Yates JR III: An automated multidi-
mensional protein identification technology for shotgun pro-
teomics. Anal Chem 2001, 73:5683-5690. 3. Gan CS, Chong PK, Pham TK, Wright PC: Technical, experimen-
tal, and biological variations in isobaric tags for relative and
absolute quantitation (iTRAQ). J Proteome Res 2007, 6:821-827. q
(
)
J
4. Pierce A, Unwin RD, Evans CA, Griffiths S, Carney L, Zhang L, Jawor-
ska E, Lee CF, Blinco D, Okoniewski MJ, et al.: Eight-channel
iTRAQ enables comparison of the activity of 6 leu-
kaemogenic tyrosine kinases. Mol Cell Proteomics 2007, 7:853-83. 5. Wolters DA, Washburn MP, Yates JR III: An automated multidi-
mensional protein identification technology for shotgun pro-
teomics. Anal Chem 2001, 73:5683-5690. 6. 6. Essader AS, Cargile BJ, Bundy JL Jr: A comparison of immobilized
pH gradient isoelectric focusing and strong-cation-exchange
chromatography as a first dimension in shotgun proteomics. Proteomics 2005, 5:24-34. The experimental pI for each peptide was calculated using
a pI/Mw tool of the ExPASy Proteomic Server [12,29]. 7. Competing interests p
g
The authors declare that they have no competing interests. ,
10. Michel PE, Reymond F, Arnaud I, Josserand J, Girault HH, Rossier JS:
Protein fractionation in a multicompartment device using
Off-Gel isoelectric focusing. Electrophoresis 2003, 24:3-11. Additional material signals per spot position having a S/N > 12 were selected
as precursors for MS/MS acquisition. Peptide and protein
identification were performed by the ProteinPilot™ Soft-
ware V 2.0 (Applied Biosystems) using the Paragon algo-
rithm [28]. Each MS/MS spectrum was searched for Homo
sapiens specie against the Uniprot/swissprot database
(release 51 of October 2006). The searches were run using
with the fixed modification of methylmethanethiosulfate
labelled cysteine parameter enabled. Other parameters
such as tryptic cleavage specificity, precursor ion mass
accuracy and fragment ion mass accuracy are MALDI 4800
built-in functions of ProteinPilot software. Additional file 1
List of identified proteins with at least 2 peptides. The proteins provided
result from the combination of all the experiements. Click here for file
[http://www.biomedcentral.com/content/supplementary/1477-
5956-6-27-S1.pdf] Additional file 1
List of identified proteins with at least 2 peptides. The proteins provided
result from the combination of all the experiements. Click here for file
[http://www.biomedcentral.com/content/supplementary/1477-
5956-6-27-S1.pdf] Additional file 1
List of identified proteins with at least 2 peptides. The proteins provided
result from the combination of all the experiements. Click here for file Additional file 1
List of identified proteins with at least 2 peptides. The proteins provided
result from the combination of all the experiements. Click here for file [http://www.biomedcentral.com/content/supplementary/1477-
5956-6-27-S1.pdf] [http://www.biomedcentral.com/content/supplementary/1477-
5956-6-27-S1.pdf] References Heller M, Michel PE, Morier P, Crettaz D, Wenz C, Tissot JD, Rey-
mond F, Rossier JS: Two-stage Off-Gel isoelectric focusing: pro-
tein followed by peptide fractionation and application to
proteome analysis of human plasma. Electrophoresis 2005,
26:1174-1188. Abbreviations NHS: N-Hydrosuccinimide; SCX: strong cation exchange;
IEF: isoelectric focusing; IPG: Immobilized pH gradient;
MMTS: methyl methanethiosulfonate; TFA: trifluoroacetic
acid; PAI: protein abundance indice; CHCA: α-cyano-4-
hydroxy-cinnamic acid; ACN: acetonitrile. 8. Michel PE, Crettaz D, Morier P, Heller M, Gallot D, Tissot JD, Rey-
mond F, Rossier JS: Proteome analysis of human plasma and
amniotic fluid by Off-Gel isoelectric focusing followed by
nano-LC-MS/MS. Electrophoresis 2006, 27:1169-1181. p
9. Scherl A, Francois P, Charbonnier Y, Deshusses JM, Koessler T, Huy-
ghe A, Bento M, Stahl-Zeng J, Fischer A, et al.: Exploring glycopep-
tide-resistance in Staphylococcus aureus: a combined
proteomics and transcriptomics approach for the identifica-
tion of resistance-related markers. BMC Genomics 2006, 7:296. Acknowledgements
Th
k
d b This work was supported by grants from the "Ligue Nationale Contre le
Cancer" (program équipe labellisée). The ProteinPilot software calculates a confidence percent-
age (the unused score) which reflects the probability that
the hit is a false positive, meaning that at 95% confidence
level, there is a false positive identification chance of
about 5%. Peptide OFFGEL fractionation MS and MS/MS analyses of off-line spotted peptide sam-
ples were performed using the 4800 MALDI-TOF/TOF
Analyser (Applied Biosystems). After screening of all LC-
MALDI sample positions in MS-positive reflector mode
using 1500 laser shots, the fragmentation of automati-
cally-selected precursors was performed at collision
energy of 1 kV using air as collision gas (pressure of ~2 ×
10-6 Torr). MS spectra were acquired between m/z 800 and
4000. For internal calibration, we used the parent ion of
Glu1-fibrinopeptide at m/z 1570.677 diluted in the matrix
(3 femtomoles per spot). Up to 12 of the most intense ion p
f
For pI-based peptide separation, we used the 3100 OFF-
GEL Fractionator (Agilent Technologies, Böblingen, Ger-
many) with a 24-well set-up. Prior to electrofocusing,
samples were desalted onto a Sep-Pak C18 cartridge
(Waters). For 24-well set-up, peptide samples were diluted
to a final volume of respectively 3.6 mL using OFFGEL
peptide sample solution. To start, the IPG gel strips of 24
cm-long (GE Healthcare, München, Germany) with a 3–
10 linear pH range were rehydrated with the Peptide IPG
Strip Rehydradation Solution according to the protocol of Page 11 of 13
(page number not for citation purposes) Page 11 of 13
(page number not for citation purposes) Proteome Science 2008, 6:27 http://www.proteomesci.com/content/6/1/27 Page 12 of 13
(page number not for citation purposes) Authors' contributions EE performed the laboratory experiments, carried out the
LC separation of peptides samples and assisted in the writ-
ing of the final manuscript. EG supervised the project. CG
carried out the MALDI-TOF/TOF analysis, participated in
the design of the study and wrote the manuscript. CG
supervised and coordinated the project. All authors read
and approved the final manuscript. 11. Burgess JA, Lescuyer P, Hainard A, Burkhard PR, Turck N, Michel P,
Rossier JS, Reymond F, Hochstrasser DF, Sanchez JC: Identification
of brain cell death associated proteins in human post-mor-
tem cerebrospinal fluid. J Proteome Res 2006, 5:1674-1681. 12. Bjellgvist B, Hughes GJ, Pasquali C, Paquet N, Ravier F, Sanchez JC,
Frutiger S, Hochstrasser D: The focusing positions of polypep-
tides in immobilized pH gradients can be predicted from
their amino acid sequences. Electrophoresis 1993, 14:1023-1031. q
p
13. Krügsveld J, Gauci S, Dormever W, Heck AJ: In-gel isoelectric
focusing of peptides as a tool for improved protein identifica-
tion. J Proteome Res 2006, 5:1721-1730. J
14. Cargile BJ, Bundy JL, Freeman TW, Stephenson JL Jr: Immobilized
pH gradients as a first dimension in shotgun proteomics and
analysis of accuracy of pI predictability of peptides. J Proteome
Res 2004, 3:112-119. 14. Cargile BJ, Bundy JL, Freeman TW, Stephenson JL Jr: Immobilized
pH gradients as a first dimension in shotgun proteomics and
analysis of accuracy of pI predictability of peptides. J Proteome
Res 2004, 3:112-119. Page 12 of 13
(page number not for citation purposes) Proteome Science 2008, 6:27 http://www.proteomesci.com/content/6/1/27 http://www.proteomesci.com/content/6/1/27 15. Lengqvist J, Uhlen K, Lehtio J: iTRAQ compatibility of peptide
immobilized pH gradient isoelectric focusing. Proteomics 2007,
7:1746-1752. 16. Chenau J, Michelland S, Sidibe J, Seve M: Peptides OFFGEL elec-
trophoresis: a suitable pre-analytical step for complex
eukaryotic samples fractionation compatible with quantita-
tive iTRAQ labeling. Proteome Sci 2008:6-9. 17. Constantinescu R, Constantinescu AT, Reichmann H, Janetzky B:
Neuronal differentiation and long-term culture of the
human neuroblastoma line SH-SY5Y. J Neural Transm Suppl
2007, 72:17-28. 18. Villa D, Miloso M, Nicolini G, Rigolio R, Villa A, Cavaletti G, Tredici
G: Low-dose cisplatin protects human neuroblastoma SH-
SY5Y cells from paclitaxel-induced apoptosis. Mol Cancer Ther
2005, 4:1439-1447. 19. Gilany K, van Elzen R, Mous K, Coen E, van Dongen W, Vandamme
S, Gevaert K, Timmerman E, Vandekerckhove J, Dewilde S, et al.: The
proteome of the human neuroblastoma cell line SH-SY5Y:
an enlarged proteome. Biochim Biophys Acta 2008, 1784:983-5. Authors' contributions g
p
p y
20. Nishikaze T, Takayama M: Cooperative effect of factors govern-
ing molecular ion yields in desorption/ionization mass spec-
trometry. Rapid Commun Mass Spectrom 2006, 20(3):376-382. y
p
p
( )
21. Hörth P, Miller CA, Preckel T, Wenz C: Efficient fractionation
and improved protein identification by peptide OFFGEL
electrophoresis. Mol Cell Proteomics 2006, 5:1968-1974. p
22. Cargile BJ, Talley DL, Stephenson JL: Immobilized pH gradients
as a first dimension in shotgun proteomics and analysis of the
accuracy of pI predictability of peptides. Electrophoresis 2004,
25:936-945. 23. Fraterman S, Zeiger U, Khurana TS, Rubinstein NA, Wilm M: Com-
bination of peptide OFFGEL fractionation and label-free
quantitation facilitated proteomics profiling of extraocular
muscle. Proteomics 2007, 7:3404-3416. 24. Cargile BJ, Bundy JL, Stephenson JL Jr: Potential for false positive
identification from large databases through tandem mass
spectrometry. J Proteome Res 2004, 3:1082-1085. p
y
J
25. Liu T, Qian WJ, Chen WN, Jacobs JM, Moore RJ: Improved pro-
teome coverage by using high efficiency cysteinyl peptide
enrichment: the human mammary epithelial cell proteome. Proteomics 2005, 5:1263-1273. 26. Schwartz R, Ting CS, King J: Whole proteome pI values corre-
late with subcellular localizations of proteins for organisms
within the three domains of life. Genome Res 2001,
11(5):703-709. ( )
27. Rappsilber J, Ryder U, Lamond AI, Mann M: Large-scale proteomic
analysis of the human spliceosome. Genome Res 2002,
12:1231-1245. 28. Shilov IV, Seymour SL, Patel AA, Loboda A, Tang WH, Keating SP,
Hunter CL, Nuwaysir LM, Schaeffer DA, et al.: The Paragon Algo-
rithm, a next generation search engine that uses sequence
temperature values and feature probabilities to identify pep-
tides from tandem mass spectra. Mol Cell Proteomics 2007,
6:1638-1655. 29. Gasteiger E, Gattiker A, Hoogland C, Ivanyi I, Appel RD, Bairoch A,
et al.: ExPASy: The proteomics server for in-depth protein
knowledge and analysis. Nucleic Acids Res 2003, 31:3784-3788. Authors' contributions Publish with BioMed Central and every
scientist can read your work free of charge
"BioMed Central will be the most significant development for
disseminating the results of biomedical research in our lifetime."
Sir Paul Nurse, Cancer Research UK
Your research papers will be:
available free of charge to the entire biomedical community
peer reviewed and published immediately upon acceptance
cited in PubMed and archived on PubMed Central
yours — you keep the copyright
Submit your manuscript here:
http://www.biomedcentral.com/info/publishing_adv.asp
BioMedcentral Publish with BioMed Central and every
scientist can read your work free of charge
"BioMed Central will be the most significant development for
disseminating the results of biomedical research in our lifetime."
Sir Paul Nurse, Cancer Research UK
Your research papers will be:
available free of charge to the entire biomedical community
peer reviewed and published immediately upon acceptance
cited in PubMed and archived on PubMed Central
yours — you keep the copyright
Submit your manuscript here:
http://www.biomedcentral.com/info/publishing_adv.asp
BioMedcentral Publish with BioMed Central and every
scientist can read your work free of charge Page 13 of 13
(page number not for citation purposes)
|
https://openalex.org/W2341598274
|
https://hal-uphf.archives-ouvertes.fr/hal-03445985/document
|
English
| null |
Energetic and Exergetic Analysis of a Heat Exchanger Integrated in a Solid Biomass-Fuelled Micro-CHP System with an Ericsson Engine
|
Entropy
| 2,016
|
cc-by
| 14,135
|
To cite this version: Marie Creyx, Eric Delacourt, Céline Morin, Sylvain Lalot, Bernard Desmet. Energetic and Exergetic
Analysis of a Heat Exchanger Integrated in a Solid Biomass-Fuelled Micro-CHP System with an
Ericsson Engine. Entropy, 2016, 18 (4), pp.154. 10.3390/e18040154. hal-03445985 Distributed under a Creative Commons Attribution 4.0 International License Marie Creyx 1, Eric Delacourt 1, Céline Morin 1,*, Sylvain Lalot 1 and Bernard Desmet 2 1
LAMIH (UMR CNRS 8201), University of Valenciennes (UVHC), Le Mont Houy,
F-59313 Valenciennes Cedex 9, France; marie.creyx@gmail.com (M.C.);
eric.delacourt@univ-valenciennes.fr (E.D.); sylvain.lalot@univ-valenciennes.fr (S.L.)
2
ENSIAME (Engineering School), University of Valenciennes (UVHC), Le Mont Houy,
F-59313 Valenciennes Cedex 9, France; bernard.desmet@univ-valenciennes.fr
*
Correspondence: celine.morin@univ-valenciennes.fr; Tel.: +33-3-27-511964; Fax: +33-3-27-511961
Academic Editor: Kevin H. Knuth 1
LAMIH (UMR CNRS 8201), University of Valenciennes (UVHC), Le Mont Houy,
F-59313 Valenciennes Cedex 9, France; marie.creyx@gmail.com (M.C.);
eric.delacourt@univ-valenciennes.fr (E.D.); sylvain.lalot@univ-valenciennes.fr (S.L.)
2
ENSIAME (Engineering School), University of Valenciennes (UVHC), Le Mont Houy,
F-59313 Valenciennes Cedex 9, France; bernard.desmet@univ-valenciennes.fr
*
Correspondence: celine.morin@univ-valenciennes.fr; Tel.: +33-3-27-511964; Fax: +33-3-27-511961 1
LAMIH (UMR CNRS 8201), University of Valenciennes (UVHC), Le Mont Houy,
F-59313 Valenciennes Cedex 9, France; marie.creyx@gmail.com (M.C.);
eric.delacourt@univ-valenciennes.fr (E.D.); sylvain.lalot@univ-valenciennes.fr (S.L.)
2
ENSIAME (Engineering School), University of Valenciennes (UVHC), Le Mont Houy,
F-59313 Valenciennes Cedex 9, France; bernard.desmet@univ-valenciennes.fr
*
Correspondence: celine.morin@univ-valenciennes.fr; Tel.: +33-3-27-511964; Fax: +33-3-27-511961
Academic Editor: Kevin H. Knuth 1
LAMIH (UMR CNRS 8201), University of Valenciennes (UVHC), Le Mont Houy,
F-59313 Valenciennes Cedex 9, France; marie.creyx@gmail.com (M.C.);
eric.delacourt@univ-valenciennes.fr (E.D.); sylvain.lalot@univ-valenciennes.fr (S.L.)
2
ENSIAME (Engineering School), University of Valenciennes (UVHC), Le Mont Houy, Academic Editor: Kevin H. Knuth Received: 22 October 2015; Accepted: 12 April 2016; Published: 20 April 2016 Received: 22 October 2015; Accepted: 12 April 2016; Published: 20 April 2016 Abstract: A specific heat exchanger has been developed to transfer heat from flue gas to the working
fluid (hot air) of the Ericsson engine of a solid biomass-fuelled micro combined heat and power
(CHP). In this paper, the theoretical and experimental energetic analyses of this heat exchanger
are compared. The experimental performances are described considering energetic and exergetic
parameters, in particular the effectiveness on both hot and cold sides. A new exergetic parameter
called the exergetic effectiveness is introduced, which allows a comparison between the real and the
ideal heat exchanger considering the Second Law of Thermodynamics. A global analysis of exergetic
fluxes in the whole micro-CHP system is presented, showing the repartition of the exergy destruction
among the components. Keywords:
micro-CHP system; biomass; heat exchanger; energetic and exergetic analysis;
exergetic effectiveness Entropy 2016, 18, 154; doi:10.3390/e18040154 HAL Id: hal-03445985
https://uphf.hal.science/hal-03445985v1
Submitted on 25 Apr 2022 L’archive ouverte pluridisciplinaire HAL, est
destinée au dépôt et à la diffusion de documents
scientifiques de niveau recherche, publiés ou non,
émanant des établissements d’enseignement et de
recherche français ou étrangers, des laboratoires
publics ou privés. HAL is a multi-disciplinary open access
archive for the deposit and dissemination of sci-
entific research documents, whether they are pub-
lished or not. The documents may come from
teaching and research institutions in France or
abroad, or from public or private research centers. Distributed under a Creative Commons Attribution 4.0 International License entropy entropy Energetic and Exergetic Analysis of a Heat Exchanger
Integrated in a Solid Biomass-Fuelled Micro-CHP
System with an Ericsson Engine Marie Creyx 1, Eric Delacourt 1, Céline Morin 1,*, Sylvain Lalot 1 and Bernard Desmet 2 1. Introduction In the current global context, with growing energy consumption, climate change linked
to greenhouse gas emissions and limited fossil energy resources, solid biomass-fuelled micro
combined heat and electrical power (micro-CHP) systems offer several benefits. In particular, the
micro-CHP concept allows primary energy savings (high global efficiency is reached) compared with
a separate production of heat and electrical power. The development of such systems also represents
a decentralization of the energy production process that limits electrical power transmission and
distribution losses. Solid biomass-fuelled micro-CHP systems are composed of a solid biomass-fuelled boiler and
a cogeneration device supplied by means of a heat exchanger to produce mechanical or electrical energy
(cf. Figure 1). The literature mentions several solid biomass-fuelled micro-CHP systems associated
to various cogeneration devices: a Stirling hot air engine [1–4], an Organic Rankine Cycle (ORC)
turbine [5] and a thermoelectric generator [6]. Although no study has dealt with the performances
of a solid biomass-fuelled micro-CHP system with an Ericsson hot air engine yet, this cogeneration
device is well adapted in such facilities [7]. The Ericsson engine thermal efficiency can reach 52% [8]. Contrary to other hot air engines, the Ericsson engine does not necessarily require a cold source as
it can work with an open cycle as chosen in the present study. In this case, low pressure levels are
needed, leading to a less compact engine. Entropy 2016, 18, 154; doi:10.3390/e18040154 www.mdpi.com/journal/entropy 2 of 17 Entropy 2016, 18, 154 Figure 1. Configuration of solid biomass-fuelled micro-CHP systems. Figure 1. Configuration of solid biomass-fuelled micro-CHP systems. Figure 1. Configuration of solid biomass-fuelled micro-CHP systems. Figure 1. Configuration of solid biomass-fuelled micro-CHP systems. A biomass-fuelled micro-CHP system with an open cycle Ericsson engine composed of a
compression cylinder and an expansion cylinder requires the integration of a specific heat exchanger
in the biomass-fuelled boiler to provide hot air to the engine. This heat exchanger encounters strong
constraints. The flue gas from biomass combustion presents high temperatures (500 to 1000 °C
depending on the boiler working conditions) and contains soot particles (1 to 5 ppm). The volume of
the combustion chamber in the boiler is reduced, limiting the volume of the heat exchanger. The air
supplying the engine is pressurized (pressure range under 10 bar corresponding to the optimal
working conditions of the Ericsson engine). 1. Introduction The pressure drops have to be limited on the flue gas
side to limit the impact on biomass combustion and on the air side to ensure the high performance of
the Ericsson engine
A biomass-fuelled micro-CHP system with an open cycle Ericsson engine composed of a compression
cylinder and an expansion cylinder requires the integration of a specific heat exchanger in the
biomass-fuelled boiler to provide hot air to the engine. This heat exchanger encounters strong constraints. The flue gas from biomass combustion presents high temperatures (500 to 1000 ˝C depending on the
boiler working conditions) and contains soot particles (1 to 5 ppm). The volume of the combustion
chamber in the boiler is reduced, limiting the volume of the heat exchanger. The air supplying the
engine is pressurized (pressure range under 10 bar corresponding to the optimal working conditions
of the Ericsson engine). The pressure drops have to be limited on the flue gas side to limit the impact
on biomass combustion and on the air side to ensure the high performance of the Ericsson engine. the Ericsson engine. The gas-gas heat exchangers adapted to flue gas from biomass combustion mentioned in the
literature are recuperators supplying micro gas turbines with external combustion [9–11] or tubular
heat exchangers associated with Stirling engines [4,12,13]. Neither configuration is suited for
Ericsson engine supply. The recuperators are designed for high power systems compared with the
power required for micro-CHP systems. The tubular heat exchangers developed for Stirling engine
supply are very compact and would limit the heat transfer and the performances in the case of a
micro-CHP unit with an Ericsson engine. Among high temperature heat exchanger technologies,
the multi-tubular heat exchanger with separated tubes appears the most appropriate considering all
The gas-gas heat exchangers adapted to flue gas from biomass combustion mentioned in the
literature are recuperators supplying micro gas turbines with external combustion [9–11] or tubular
heat exchangers associated with Stirling engines [4,12,13]. Neither configuration is suited for Ericsson
engine supply. The recuperators are designed for high power systems compared with the power
required for micro-CHP systems. The tubular heat exchangers developed for Stirling engine supply
are very compact and would limit the heat transfer and the performances in the case of a micro-CHP
unit with an Ericsson engine. Among high temperature heat exchanger technologies, the multi-tubular
heat exchanger with separated tubes appears the most appropriate considering all the constraints. 1. Introduction the multi-tubular heat exchanger with separated tubes appears the most appropriate considering all
the constraints. To determine the working conditions and the performances of the solid biomass-fuelled
micro-CHP systems and of their components, an energetic analysis is mostly applied [1,2,5,6]. Such
studies may be completed with exergetic analyses, following the example presented by [14] with a
natural gas micro-CHP system, in order to highlight the zones of exergy destruction and allow an
improved optimization of the working conditions to enhance the performances. This approach can
be applied in particular to the study of the heat exchanger supplying the Ericsson engine associated
to the solid biomass-fuelled micro-CHP system considered. An energetic study of the heat transfer in
this heat exchanger performed with a numerical modelling including the phenomena of conduction
To determine the working conditions and the performances of the solid biomass-fuelled
micro-CHP systems and of their components, an energetic analysis is mostly applied [1,2,5,6]. Such studies may be completed with exergetic analyses, following the example presented by [14]
with a natural gas micro-CHP system, in order to highlight the zones of exergy destruction and allow
an improved optimization of the working conditions to enhance the performances. This approach can
be applied in particular to the study of the heat exchanger supplying the Ericsson engine associated to
the solid biomass-fuelled micro-CHP system considered. An energetic study of the heat transfer in
this heat exchanger, performed with a numerical modelling including the phenomena of conduction,
convection and flue gas radiation, is described in [15]. this heat exchanger, performed with a numerical modelling including the phenomena of conduction,
convection and flue gas radiation, is described in [15]. The objectives of the present work are firstly to validate the energetic study [15] with
experimental results and secondly to present an exergetic analysis of the heat exchanger inserted in
The objectives of the present work are firstly to validate the energetic study [15] with experimental
results, and secondly to present an exergetic analysis of the heat exchanger inserted in the solid
biomass-fuelled micro-CHP system. experimental results, and secondly to present an exergetic analysis of the heat exchanger inserted in
the solid biomass-fuelled micro-CHP system. This paper is structured as follows: first, the configuration of the heat exchanger implemented
in the micro-CHP system is described. The instrumentation and the experimental methodology are
then specified. 2. Configuration of the Heat Exchanger Associated to the Biomass-Fuelled Micro-CHP Unit
2. Configuration of the Heat Exchanger Associated to the Biomass-Fuelled Micro-CHP Unit The heat exchanger supplying the Ericsson engine of the biomass-fuelled micro-CHP unit is
composed of metallic vertical tubes linked by spacers to form a cylinder (cf. Figure 2). It is a 1-2 heat
exchanger: the flue gas flows inside the cylinder (one pass) while the air flows in the tubes (two passes). The heat exchanger is inserted in a domestic biomass-fuelled boiler, in the combustion chamber and
above the furnace. The inlet air of the heat exchanger is previously compressed in the compression
cylinder of the Ericsson engine. The exhaust air is linked to the expansion cylinder of the Ericsson
engine [7]. It has been specifically designed to fit the dimensions of the boiler. It consists of 16 tubes of
21.5 inner diameter, 2 mm thick, 20 cm long, located on a 32 cm diameter circle. They are linked by fins
that are welded to the tubes. The whole exchanger is made of 310S stainless steel. The heat exchanger supplying the Ericsson engine of the biomass-fuelled micro-CHP unit is
composed of metallic vertical tubes linked by spacers to form a cylinder (cf. Figure 2). It is a 1-2 heat
exchanger: the flue gas flows inside the cylinder (one pass) while the air flows in the tubes (two
passes). The heat exchanger is inserted in a domestic biomass-fuelled boiler, in the combustion
chamber and above the furnace. The inlet air of the heat exchanger is previously compressed in the
compression cylinder of the Ericsson engine. The exhaust air is linked to the expansion cylinder of
the Ericsson engine [7]. It has been specifically designed to fit the dimensions of the boiler. It consists
of 16 tubes of 21.5 inner diameter, 2 mm thick, 20 cm long, located on a 32 cm diameter circle. They
are linked by fins that are welded to the tubes. The whole exchanger is made of 310S stainless steel. Figure 2. Configuration of the 1-2 heat exchanger. Figure 2. Configuration of the 1-2 heat exchanger. Figure 2. Configuration of the 1-2 heat exchanger. Figure 2. Configuration of the 1-2 heat exchanger. 1. Introduction Energetic and exergetic models of the heat exchanger inserted in the biomass-fuelled
micro-CHP facility are presented. Several results (comparison between the energetic study of the heat
exchanger and the experimental results, energetic and exergetic performances of the heat exchanger
and study of the exergetic fluxes and the exergy destruction in the micro CHP system) are finally
This paper is structured as follows: first, the configuration of the heat exchanger implemented
in the micro-CHP system is described. The instrumentation and the experimental methodology are
then specified. Energetic and exergetic models of the heat exchanger inserted in the biomass-fuelled
micro-CHP facility are presented. Several results (comparison between the energetic study of the heat
exchanger and the experimental results, energetic and exergetic performances of the heat exchanger
and study of the exergetic fluxes and the exergy destruction in the micro-CHP system) are finally
analysed and discussed. Entropy 2016, 18, 154
E t
2016 18 154 3 of 17
3
f 17 3.1. Heat Exchanger Instrumentation: Metrology and Calibration
3.1. Heat Exchanger Instrumentation: Metrology and Calibration Positions of thermocouples for the heat exchanger tests. Figure 3. Positions of thermocouples for the heat exchanger tests. Figure 3. Positions of thermocouples for the heat exchanger tests. Figure 3. Positions of thermocouples for the heat exchanger tests. The flue gas temperatures are measured with 0.2 mm diameter S type thermocouples
(measurement error of 0.25%) that can be positioned at several vertical and radial positions (cf. Figure 4). To estimate the measurement error linked to radiative flux inside the combustion chamber, the
extrapolation method is used: the temperatures are measured simultaneously with two
thermocouples of different diameters (0.2 mm and 0.13 mm). With a temperature of 600 °C without
correction, the measurement error is estimated to 44 °C and can reach 187 °C for a flue gas measured
temperature of 1000 °C (mainly because of the hot junction radiative losses). The flue gas temperatures are measured with 0.2 mm diameter S-type thermocouples
(measurement error of 0.25%) that can be positioned at several vertical and radial positions (cf. Figure 4). To estimate the measurement error linked to radiative flux inside the combustion chamber, the
extrapolation method is used: the temperatures are measured simultaneously with two thermocouples
of different diameters (0.2 mm and 0.13 mm). With a temperature of 600 ˝C without correction, the
measurement error is estimated to 44 ˝C and can reach 187 ˝C for a flue gas measured temperature of
1000 ˝C (mainly because of the hot junction radiative losses). The flue gas temperatures are measured with 0.2 mm diameter S-type thermocouples
(measurement error of 0.25%) that can be positioned at several vertical and radial positions (cf. Figure 4). To estimate the measurement error linked to radiative flux inside the combustion chamber, the
extrapolation method is used: the temperatures are measured simultaneously with two
thermocouples of different diameters (0.2 mm and 0.13 mm). With a temperature of 600 °C without
correction, the measurement error is estimated to 44 °C and can reach 187 °C for a flue gas measured
temperature of 1000 °C (mainly because of the hot junction radiative losses). Figure 4. Locations of S-type thermocouples in the combustion chamber (measurement of flue gas
temperature). The flue gas mass flow rate is estimated from the biomass fuel mass flow rate burned and from
the combustion equation. The boiler is supplied with wood pellets using a screw conveyor
Figure 4. 3.1. Heat Exchanger Instrumentation: Metrology and Calibration
3.1. Heat Exchanger Instrumentation: Metrology and Calibration Locations of S-type thermocouples in the combustion chamber (measurement of flue gas
temperature). Figure 4. Locations of S-type thermocouples in the combustion chamber (measurement of flue
gas temperature). The flue gas mass flow rate is estimated from the biomass fuel mass flow rate burned and from
he combustion equation
The boiler is supplied with wood pellets using a screw conveyor
Figure 4. Locations of S-type thermocouples in the combustion chamber (measurement of flue gas
temperature). Figure 4. Locations of S-type thermocouples in the combustion chamber (measurement of flue
gas temperature). instrumented with a magnetic sensor measuring the angular position. A calibration of the screw links
its angular rotation to the mass of pellets supplied in the boiler (cf. Figure 5). From an elementary
analysis of wood pellets, the combustion equation is established, considering an air excess λair of 0.8 [15]:
29.93
45.82
20.71
0.12
2
air
2
2
2
2
air
2
air
2
C
H
O
N
+ 3.42 H O + 31.03 1+λ
O + 0.79/0.21 N
29.93 CO + 26.33 H O + 31.03 λ
O + 31.03 1+λ
0.79/0.21 N
(1)
The flue gas mass flow rate is estimated from the biomass fuel mass flow rate burned and from
the combustion equation. The boiler is supplied with wood pellets using a screw conveyor
instrumented with a magnetic sensor measuring the angular position. A calibration of the screw links
its angular rotation to the mass of pellets supplied in the boiler (cf. Figure 5). From an elementary
analysis of wood pellets, the combustion equation is established, considering an air excess λair of 0.8 [15]:
C
H
O
N
+ 3 42 H O + 31 03 1+λ
O + 0 79/0 21 N
The flue gas mass flow rate is estimated from the biomass fuel mass flow rate burned and from the
combustion equation. The boiler is supplied with wood pellets using a screw conveyor instrumented
with a magnetic sensor measuring the angular position. A calibration of the screw links its angular
rotation to the mass of pellets supplied in the boiler (cf. Figure 5). From an elementary analysis of
wood pellets, the combustion equation is established, considering an air excess λair of 0.8 [15]: ue gas mass flow rate is deduced from Equation (1) and from the pellet mass flow rate. 3.1. Heat Exchanger Instrumentation: Metrology and Calibration
3.1. Heat Exchanger Instrumentation: Metrology and Calibration 3.1. Heat Exchanger Instrumentation: Metrology and Calibration
3.1. Heat Exchanger Instrumentation: Metrology and Calibration The heat exchanger is placed in the cylindrical combustion chamber of the biomass-fuelled
boiler. The compression cylinder of the Ericsson engine is simulated using a screw compressor
equipped with a manual valve that allows the adjustment of the air mass flow rate entering the heat
exchanger. A thermal air mass flow meter (measuring range between 0.2 and 10 Nm3/h, measurement
error of ± (0.5% reading data +0.1% full scale)) is placed at the air inlet of the heat exchanger. An
electrical air heater sets the air inlet temperature (up to 760 °C). A valve placed at the heat exchanger
outlet allows the control of the pressure level (up to 10 bar, precision of 0.04% of the full scale) in the
heat exchanger. As shown in Figure 3, the temperatures of air at the inlet, at the outlet of the heat
exchanger, and on the walls at four locations are measured with 0.3 mm diameter K-type
thermocouples (measurement error of 0.75%). From the measurements, the wall temperature can be
d
f
The heat exchanger is placed in the cylindrical combustion chamber of the biomass-fuelled boiler. The compression cylinder of the Ericsson engine is simulated using a screw compressor equipped
with a manual valve that allows the adjustment of the air mass flow rate entering the heat exchanger. A thermal air mass flow meter (measuring range between 0.2 and 10 Nm3/h, measurement error of
˘ (0.5% reading data +0.1% full scale)) is placed at the air inlet of the heat exchanger. An electrical air
heater sets the air inlet temperature (up to 760 ˝C). A valve placed at the heat exchanger outlet allows
the control of the pressure level (up to 10 bar, precision of 0.04% of the full scale) in the heat exchanger. As shown in Figure 3, the temperatures of air at the inlet, at the outlet of the heat exchanger, and on
the walls at four locations are measured with 0.3 mm diameter K-type thermocouples (measurement
error of 0.75%). From the measurements, the wall temperature can be assumed uniform. Entropy 2016, 18, 154
E
1
18 154 4 of 17 Entropy 2016, 18, 154
4 of 17
Figure 3. Positions of thermocouples for the heat exchanger tests. Figure 3. Positions of thermocouples for the heat exchanger tests. ntropy 0 6, 8, 5
o
Figure 3. 3.2. Experimental Methodology
3.2. Experimental Methodology 3.2. Experimental Methodology
3.2. Experimental Methodology Tests are conducted on the heat exchanger, considering an atmospheric air pressure and various
air inlet temperatures and volume flow rates (cf. Table 1). The boiler works in a steady-state. Each
test is carried out during 10 min after a long enough stabilization period. Tests are conducted on the heat exchanger, considering an atmospheric air pressure and various
air inlet temperatures and volume flow rates (cf. Table 1). The boiler works in a steady-state. Each test
is carried out during 10 min after a long enough stabilization period. Table 1. Experimental conditions for the heat exchanger tests. Table 1. Experimental conditions for the heat exchanger tests. Table 1. Experimental conditions for the heat exchanger tests. Parameter
Adjusted Value
Maximum Variation
during a Test of 10 min
Air volume flow rate
2.5, 5, 7.5, 10 Nm3/h
4.3%
Inlet air temperature
33, 150 °C
–
Inlet flue gas temperature
818–980 °C
9.4%
Test duration
10 min
–
Sampling frequency
1 Hz
–
4. Modelling of the Heat Exchanger Inserted in the Biomass-Fuelled Micro-CHP System
Table 1. Experimental conditions for the heat exchanger tests. Parameter
Adjusted Value
Maximum Variation
during a Test of 10 min
Air volume flow rate
2.5, 5, 7.5, 10 Nm3/h
4.3%
Inlet air temperature
33, 150 ˝C
–
Inlet flue gas temperature
818–980 ˝C
9.4%
Test duration
10 min
–
Sampling frequency
1 Hz
–
4. Modelling of the Heat Exchanger Inserted in the Biomass-Fuelled Micro-CHP System 4. Modelling of the Heat Exchanger Inserted in the Biomass-Fuelled Micro-CHP System
4. Modelling of the Heat Exchanger Inserted in the Biomass-Fuelled Micro-CHP System 3.1. Heat Exchanger Instrumentation: Metrology and Calibration
3.1. Heat Exchanger Instrumentation: Metrology and Calibration
29.93
45.82
20.71
0.12
2
air
2
2
2
2
air
2
air
2
C
H
O
N
+ 3.42 H O + 31.03 1+λ
O + 0.79/0.21 N
29.93 CO + 26.33 H O + 31.03 λ
O + 31.03 1+λ
0.79/0.21 N
(1)
fl
t i d d
d f
E
ti
(1)
d f
th
ll t
fl
t
C29.93H45.82O20.71N0.12 ` 3.42H2O ` 31.03 p1 ` λairq pO2 ` 0.79{0.21N2q
Ñ 29.93CO2 ` 26.33H2O ` 31.03λairO2 ` 31.03 p1 ` λairq p0.79{0.21q N2
(1) (1)
(1) The flue gas mass flow rate is deduced from Equation (1) and from the pellet mass flow rate. The flue gas mass flow rate is deduced from Equation (1) and from the pellet mass flow rate. 5 of 17
5 of 17 Entropy 2016, 18, 154
Entropy 2016, 18, 154 Figure 5. Calibration of the feeding screw of the biomass boiler. y = 4.529167E-04x + 5.369579E-02
R² = 9.971947E-01
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
0
2000
4000
6000
8000
supplied pellet mass (kg)
rotation angle of the feeding screw (deg)
m = 4.529·10-4·θ + 53.696·10-3
R2 = 0.997
Figure 5. Calibration of the feeding screw of the biomass boiler. Figure 5. Calibration of the feeding screw of the biomass boiler. Figure 5. Calibration of the feeding screw of the biomass boiler. 4.1. Energetic Model of the Heat Exchanger
4.1. Energetic Model of the Heat Exchanger The radiative heat transfer coefficient is evaluated by analogy with a convective heat transfer
coefficient (hypothesis of a low temperature variation of both air and flue gas flowing through the
heat exchanger): The radiative heat transfer coefficient is evaluated by analogy with a convective heat transfer
coefficient (hypothesis of a low temperature variation of both air and flue gas flowing through the
heat exchanger):
,
/
R
R
gas
wall
gas
gas
wall
h
T
T
A
T
T
(3)
with hR radiative heat transfer coefficient and Agas heat exchange surface area in contact with flue
gas. The radiative heat transfer coefficient is evaluated by analogy with a convective heat transfer
coefficient (hypothesis of a low temperature variation of both air and flue gas flowing through the
heat exchanger): s the wall radiative heat flux from flue gas depending on flue gas and wall
hR “ ϕR
`
Tgas, Twall
˘
{
`
Agas ¨
`
Tgas ´ Twall
˘˘
(3) (3)
g
temperatures,
gas
T
and
wall
T
are the flue gas and wall average temperatures. The wall radiative heat
flux that depends on flue gas and wall temperatures is evaluated from tabulated values determined
by the resolution of the Radiative Transfer Equation (RTE) with a discrete ordinate method [17],
considering the model of radiative properties of burned gases (water vapour and carbon dioxide) of
[18] and the model of radiative properties of soot particles of [17]. The wall radiative heat flux versus vertical position for several flue gas temperatures and soot
volume fractions in the range 600–1000 °C and 1–10 ppm respectively is presented in Figure 7 The
where ϕR
`
Tgas, Twall
˘
is the wall radiative heat flux from flue gas depending on flue gas and wall
temperatures, Tgas and Twall are the flue gas and wall average temperatures. The wall radiative heat
flux that depends on flue gas and wall temperatures is evaluated from tabulated values determined
by the resolution of the Radiative Transfer Equation (RTE) with a discrete ordinate method [17],
considering the model of radiative properties of burned gases (water vapour and carbon dioxide)
of [18] and the model of radiative properties of soot particles of [17]. volume fractions, in the range 600–1000 C and 1–10 ppm respectively, is presented in Figure 7. 4.1. Energetic Model of the Heat Exchanger
4.1. Energetic Model of the Heat Exchanger g
f
g
The heat transfer across the heat exchanger inserted in the solid biomass-fuelled boiler of the
micro-CHP system depends on several thermal phenomena: conduction, convection and thermal
radiation (cf. Figure 6). In particular, the flue gas from solid biomass combustion at high temperature
and containing soot particles generates an important thermal radiation and induces fouling on the
heat exchanger wall that modifies the conduction through the wall (presence of a soot deposit) [15]. The heat transfer across the heat exchanger inserted in the solid biomass-fuelled boiler of the
micro-CHP system depends on several thermal phenomena: conduction, convection and thermal
radiation (cf. Figure 6). In particular, the flue gas from solid biomass combustion at high temperature
and containing soot particles generates an important thermal radiation and induces fouling on the
heat exchanger wall that modifies the conduction through the wall (presence of a soot deposit) [15]. 6 of 17 Entropy 2016, 18, 154 Figure 6. Scheme of the heat transfer phenomena across the heat exchanger. Figure 6. Scheme of the heat transfer phenomena across the heat exchanger. Figure 6. Scheme of the heat transfer phenomena across the heat exchanger. Figure 6. Scheme of the heat transfer phenomena across the heat exchanger. (
) [
]
pp
exchanger, considering a global heat exchange coefficient evaluated from thermal resistances
(convective resistance Rcv, conductive resistance Rcd, radiative resistance RR) [15]. The radiative
resistance RR is similar to a convective resistance:
1/
R
R
gas
R
h A
(2)
The Number of Transfer Units method (NTU method) [16] is applied to model the heat exchanger,
considering a global heat exchange coefficient evaluated from thermal resistances (convective resistance
Rcv, conductive resistance Rcd, radiative resistance RR) [15]. The radiative resistance RR is similar to
a convective resistance:
g
( )
ent and Agas heat exchange surface area in contact with flue gas. RR “ 1{
`
hRAgas
˘
(2) (2) The radiative heat transfer coefficient is evaluated by analogy with a convective heat transfer
coefficient (hypothesis of a low temperature variation of both air and flue gas flowing through the
heat exchanger):
,
/
R
R
gas
wall
gas
gas
wall
h
T
T
A
T
T
(3)
with hR radiative heat transfer coefficient and Agas heat exchange surface area in contact with flue
gas. 4.1. Energetic Model of the Heat Exchanger
4.1. Energetic Model of the Heat Exchanger The equations are detailed
in the following: Rh “
.mh ch
.mc cc
(4) (4) with
.mh and
.mc hot and cold fluid mass flow rates respectively, ch and cc hot and cold fluid heat
capacities at constant pressure respectively: with
.mh and
.mc hot and cold fluid mass flow rates respectively, ch and cc hot and cold fluid heat
capacities at constant pressure respectively: Eh “
`
Th,in ´ Th,ex
˘
{
`
Th,in ´ Tc,in
˘
(5) (5) with Th,in, Th,ex the hot fluid inlet and exhaust temperatures, Tc,in the cold fluid inlet temperature Ec “
`
Tc,ex ´ Tc,in
˘
{
`
Th,in ´ Tc,in
˘
(6) Ec “
`
Tc,ex ´ Tc,in
˘
{
`
Th,in ´ Tc,in
˘ (6) with Tc,ex the cold fluid exhaust temperature: with Tc,ex the cold fluid exhaust temperature: NTU1,1-2 “
1
b
1 ` R2
1
ln
¨
˚
˚
˝1 `
b
1 ` R2
1
1
E1 ´ R1
2 ´
1`
b
1`R2
1
2
˛
‹‹‚
(7) (7) with subscript 1 corresponding to the one pass fluid. Here the flue gas is the fluid flowing with
one pass and is also the hot fluid, so that the NTU1,1-2 equals the NTUh: NTUh “ UAA{
` .mhch
˘
(8) (8) with UA the global heat transfer coefficient and A the reference heat exchange surface area: NTUc “ Rh NTUh
(9) (9) NTUc “ Rh NTUh
(9) 4.2. Exergetic Model of the Heat Exchanger Inserted in the Biomass-Fuelled Micro-CHP System The components of the micro-CHP system (cf. Figure 1) investigated in the exergetic analysis are
the following: combustion chamber (comb), heat exchanger (Ha) supplying the cogeneration system,
heat exchanger (Hw) supplying the domestic heating system, Ericsson engine used as a cogeneration
system (coge) (cf. Figure 8). The domestic heating system is not modelled in the exergetic analysis. For each component, the
exergy fluxes
. Ex linked to intake (in) and exhaust (ex) fluids, to wall heat losses and to mechanical
power transferred are evaluated. The exergy fluxes
. Ex exchanged in the micro-CHP system with the
combustion chamber (comb), the gas-air heat exchanger (Ha), the gas-water heat exchanger (Hw), the
cogeneration Ericsson engine (coge) are detailed in Figure 8, considering several phenomena: mass
transfer, heat transfer p
. Ex
. Qq and mechanical power produced p
. Ex
. Wq. 4.1. Energetic Model of the Heat Exchanger
4.1. Energetic Model of the Heat Exchanger The
wall radiative heat flux increases with temperature and soot volume fraction, with a higher influence
of the temperature. The wall radiative heat flux versus vertical position for several flue gas temperatures and soot
volume fractions, in the range 600–1000 ˝C and 1–10 ppm respectively, is presented in Figure 7. The wall radiative heat flux increases with temperature and soot volume fraction, with a higher
influence of the temperature. Entropy 2016, 18, 154
7 of 17 Figure 7. Evolution of the radiative heat flux versus the position for different flue gas temperatures
and soot volume fractions. In the heat exchanger, the hot fluid (subscript h) flowing with one pass (subscript 1) is the flue
Figure 7. Evolution of the radiative heat flux versus the position for different flue gas temperatures and
soot volume fractions. Figure 7. Evolution of the radiative heat flux versus the position for different flue gas temperatures
and soot volume fractions. I
h h
h
h h
fl
d
b
h fl
h
b
h fl
Figure 7. Evolution of the radiative heat flux versus the position for different flue gas temperatures and
soot volume fractions. Entropy 2016, 18, 154 Entropy 2016, 18, 154 7 of 17 In the heat exchanger, the hot fluid (subscript h) flowing with one pass (subscript 1) is the flue
gas. The cold fluid (subscript c) flowing with two passes (subscript 2) is air. Applying the NTU
method requires the evaluation of the heat capacity ratio Rh on the side of hot fluid, of the effectiveness
Eh and Ec on the sides of hot and cold fluids respectively, of the Number of Transfer Units of a 1-2
heat exchanger with inlets on opposite sides NTU1,1-2 and of the Number of Transfer Units of a heat
exchanger on the sides of hot and cold fluids NTUh and NTUc respectively. The domestic heatin
4.2.1. Exergetic Balances Exergetic Balances
An exergetic balance applied to the combustion part allows the evaluation of the exergy
destruction in the boiler furnace
destr
comb
Ex
:
w,
,
,
,
w,
comb
air in
biomass
gas in
a
comb
comb
Ha
comb
comb
a
gen comb
comb
A
T
Ex
Ex
Ex
Q
q
T S
T
E
(10)
with
. Exair,in
comb ,
. Exbiomass
comb
and
. Exgas,in
Ha
the exergy fluxes of intake air, biomass fuel and exhaust flue gas
of the furnace respectively,
. Qcomb and
.qcomb the thermal power and the heat flux density transmitted
through the boiler wall respectively, Ta the ambient temperature, Tw,comb the furnace wall temperature,
Aw,comb the furnace wall surface,
. Sgen,comb the entropy flux generated in the furnace. In the same way,
the exergy destruction in the heat exchangers (Ha) and (Hw), respectively
. Exdestr
Ha
and
. Exdestr
Hw , and in
the Ericsson engine
. Exdestr
coge can be evaluated from exergy balances: comb
Ex
and
,
gas in
Ha
Ex
the exergy fluxes of intake air, biomass fuel and exhaust flue gas
l
Q
d
h
h
l
d h h
fl
d
i
i
d
. Exdestr
Ha
“
´ . Exgas,in
Ha
´
. Exgas,ex
Ha
¯
´
´ . Exair,ex
Ha
´
. Exair,in
Ha
¯
(11) e gas
(11) of the furnace respectively,
comb
Q
and
comb
q
the thermal power and the heat flux density transmitted
through the boiler wall respectively, Ta the ambient temperature, Tw,comb the furnace wall temperature,
Aw,comb the furnace wall surface,
,
gen comb
S
the entropy flux generated in the furnace. with
. Exgas,in
Ha
and
. Exgas,ex
Ha
the exergy fluxes of the intake and exhaust flue gas of the heat exchanger Ha
respectively,
. Exair,in
Ha
and
. Exair,ex
Ha
the exergy fluxes of the intake and exhaust air of the heat exchanger
Ha, respectively: exergy destruction in the heat exchangers (Ha) and (Hw), respectively
destr
Ha
Ex
Ericsson engine
destr
Ex
can be evaluated from exergy balances:
. Exdestr
Hw “
´ . Exgas,ex
Ha
´
. Exgas,ex
Hw
¯
´
´ . Exwater,ex
Hw
´
. The domestic heatin
4.2.1. Exergetic Balances exergy fluxes Ex linked to intake (in) and exhaust (ex) fluids, to wall heat losses and to mechanical
power transferred are evaluated. The exergy fluxes Ex
exchanged in the micro-CHP system with
the combustion chamber (comb) the gas-air heat exchanger (Ha) the gas-water heat exchanger (Hw)
An exergetic balance applied to the combustion part allows the evaluation of the exergy
destruction in the boiler furnace
. Exdestr
comb: (
),
g
g
(
),
g
g
(
),
neration Ericsson engine (coge) are detailed in Figure 8, considering several phenomena:
nsfer, heat transfer (
)
Q
Ex
and mechanical power produced (
)
W
Ex
. The fluids (air, biomass
, water) are indicated in the following figure for the intake (in) and the exhaust (ex) of each
. Exair,in
comb `
. Exbiomass
comb
´
. Exgas,in
Ha
“ ´
¨
˝
. Qcomb ´
x
Aw,comb
Ta
Tw,comb
δ
.qcomb
˛
‚´ Ta
. Sgen,comb
loooooooooooooooooooooooooooooooomoooooooooooooooooooooooooooooooon
. Exdestr
comb
(10) (
),
g
g
(
),
g
g
(
),
neration Ericsson engine (coge) are detailed in Figure 8, considering several phenomena:
nsfer, heat transfer (
)
Q
Ex
and mechanical power produced (
)
W
Ex
. The fluids (air, biomass
water) are indicated in the following figure for the intake (in) and the exhaust (ex) of each
. Exair,in
comb `
. Exbiomass
comb
´
. Exgas,in
Ha
“ ´
¨
˝
. Qcomb ´
x
Aw,comb
Ta
Tw,comb
δ
.qcomb
˛
‚´ Ta
. Sgen,comb
loooooooooooooooooooooooooooooooomoooooooooooooooooooooooooooooooon
. Exdestr
comb
(10) (
)
g
g
(
)
g
g
(
)
neration Ericsson engine (coge) are detailed in Figure 8, considering several phenomena:
nsfer, heat transfer (
)
Q
Ex
and mechanical power produced (
)
W
Ex
. The fluids (air, biomass
water) are indicated in the following figure for the intake (in) and the exhaust (ex) of each
. Exair,in
comb `
. Exbiomass
comb
´
. Exgas,in
Ha
“ ´
¨
˝
. Qcomb ´
x
Aw,comb
Ta
Tw,comb
δ
.qcomb
˛
‚´ Ta
. Sgen,comb
loooooooooooooooooooooooooooooooomoooooooooooooooooooooooooooooooon
. Exdestr
comb
(10) mass
(10) 4.2.1. 4.1. Energetic Model of the Heat Exchanger
4.1. Energetic Model of the Heat Exchanger The fluids (air, biomass fuel, gas,
water) are indicated in the following figure for the intake (in) and the exhaust (ex) of each element. 8 of 17
eration Entropy 2016, 18, 154
heat exchanger (H
system (coge) (cf F 8 of 17
eration Figure 8. Exergy exchanges in the solid biomass-fuelled micro-CHP system. Figure 8. Exergy exchanges in the solid biomass-fuelled micro-CHP system. Figure 8. Exergy exchanges in the solid biomass-fuelled micro-CHP system. Figure 8. Exergy exchanges in the solid biomass-fuelled micro-CHP system. 4.2.2. Exergy Flux of Solid Biomass Fuel Contrary to the exergy fluxes of air and flue gas (that can be calculated with the specific enthalpy
and specific entropy variations of the gases from reference conditions [19]), the exergy flux of a fuel
requires the evaluation of its specific chemical exergy (thermal exergy released by chemical reactions
until reaching an equilibrium state compared with the chemical composition of the environment),
which is linked to the chemical process of combustion. For solid biomass fuels, the specific chemical exergy cannot be evaluated from thermodynamic
equations, as for common gas and liquid fuels, due to a lack of the necessary thermodynamic data
(specific enthalpy, entropy and chemical potential of solid biomass fuels depending on thermodynamic
conditions). Several methods are mentioned in the literature to evaluate the specific chemical exergy
of solid biomass fuels [20–24]. They are based on experimental correlations taking into account
the lower heating value of the biomass fuel [20–24], the elementary composition [20,24], the ash
quantity [20,23,24], the humidity of the fuel [23], the presence of sulphur [23]. For the wood pellets used
here, an experimental elementary analysis of the composition including the humidity (cf. Equation (1))
and an experimental evaluation of the lower heating value (17,900 kJ/kg) are established. In this study,
the specific chemical exergy of pellets considered is evaluated with the correlation of Szargut [23] and
reaches 20,495 kJ/kg. 4.2.3. Exergetic Performances of the Heat Exchanger Ha: Exergetic Efficiency and Exergetic Effectiveness 4.2.3. Exergetic Performances of the Heat Exchanger Ha: Exergetic Efficiency and Exergetic Effectiveness
The exergetic efficiency of the heat exchanger is evaluated as follows: 3. Exergetic Performances of the Heat Exchanger Ha: Exergetic Efficiency and Exergetic Effectiveness The exergetic efficiency of the heat exchanger is evaluated as follows: The exergetic efficiency of the heat exchanger is evaluated as follows: ηex,Ha “
. Exair,ex
Ha
´
. Exair,in
Ha
. Exgas,in
Ha
´
. Exgas,ex
Ha
(14) (14) It corresponds to the ratio between the exergetic flux received by the cold fluid and the exergetic
flux lost by the hot fluid. It corresponds to the ratio between the exergetic flux received by the cold fluid and the exergetic
flux lost by the hot fluid. In the NTU method, the real heat exchanger is compared to an ideal heat exchanger corresponding
to the case where the hot fluid exhaust temperature equals the cold fluid inlet temperature (effectiveness
evaluated on the side of hot fluid, cf. The domestic heatin
4.2.1. Exergetic Balances Exwater,in
Hw
¯
(12) destr
Ha
Ex
(12) Hw
g
coge
gy
gas,
gas,ex
,ex
,
destr
in
air
air in
Ha
Ha
Ha
Ha
Ha
Ex
Ex
Ex
Ex
Ex
(11)
with
. Exgas,ex
Ha
the exergy flux of the flue gas entering the heat exchanger Hw (coming from the exhaust
of the heat exchanger Ha),
. Exgas,ex
Hw
the exergy flux of the flue gas at the exhaust of the heat exchanger
Hw,
. Exwater,in
Hw
and
. Exwater,ex
Hw
the exergy fluxes of the intake and exhaust water of the heat exchanger
Hw, respectively: Entropy 2016, 18, 154 9 of 17 . Exair,in
coge ´
. Exair,in
Ha
`
. Exair,ex
Ha
´
. Exair,ex
coge ´
. Ex
. W
coge “ ´
¨
˝
. Qcoge ´
x
Aw,coge
Ta
Tw,coge
δ
.qcoge
˛
‚´ Ta
. Sgen,coge
loooooooooooooooooooooooooooooomoooooooooooooooooooooooooooooon
. Exdestr
coge
(13) . Exair,in
coge ´
. Exair,in
Ha
`
. Exair,ex
Ha
´
. Exair,ex
coge ´
. Ex
. W
coge “ ´
¨
˝
. Qcoge ´
x
Aw,coge
Ta
Tw,coge
δ
.qcoge
˛
‚´ Ta
. Sgen,coge
loooooooooooooooooooooooooooooomoooooooooooooooooooooooooooooon
. Exdestr
coge
(13) (13) m
. Exdestr
coge with
. Exair,in
coge the exergy flux of the intake air of the Ericsson engine from the atmosphere,
. Exair,in
Ha
the
exergy flux of compressed air leaving the Ericsson engine and entering the air heat exchanger Ha,
. Exair,ex
Ha
the exergy flux entering the Ericsson engine and coming from the exhaust of the heat exchanger
Ha,
. Exair,ex
coge the exergy flux at the exhaust of the Ericsson engine,
. Ex
. W
comb the exergy flux linked to
effective mechanical power produced,
. Qcoge and
.qcoge the thermal power and the heat flux density
transmitted through the wall of the Ericsson engine respectively, Tw,coge the wall temperature of the
Ericsson engine, Aw,coge the wall surface of the Ericsson engine,
. Sgen,coge the entropy flux generated in
the Ericsson engine. The exergy destruction
. Exdestr evaluated in Equations (10)–(13) is then expressed with two terms. The first term is linked to a heat transfer through the wall of the component considered, the second
term is linked to generated entropy. 4.2.2. Exergy Flux of Solid Biomass Fuel This exergetic effectiveness corresponds to the real gain of the exergetic flux from the cold fluid,
i
h
i ll
i
f h
h
i
l
h
b
li d b
h
ld flid
d with hc and sc respectively specific enthalpy and entropy of cold fluid, Tc,ex exhaust cold fluid temperature. with hc and sc respectively specific enthalpy and entropy of cold fluid, Tc,ex exhaust cold fluid temperature. This exergetic effectiveness corresponds to the real gain of the exergetic flux from the cold fluid,
i.e., the potentially gain of the mechanical power that can be supplied by the cold fluid, compared to
the gain that could be obtained by taking the cold fluid to the hot fluid inlet temperature. This exergetic effectiveness corresponds to the real gain of the exergetic flux from the cold fluid,
i.e., the potentially gain of the mechanical power that can be supplied by the cold fluid, compared to
the gain that could be obtained by taking the cold fluid to the hot fluid inlet temperature. 4.2.2. Exergy Flux of Solid Biomass Fuel Equation (5)) or to the case where the cold fluid outlet
temperature reaches the hot fluid inlet temperature (effectiveness evaluated on the side of cold fluid, Entropy 2016, 18, 154 10 of 17 cf. Equation (6)), considering the First Law of Thermodynamics. A similar comparison between real
and ideal heat exchangers can be applied considering the Second Law of Thermodynamics. The ideal
heat exchanger is defined, for the hot side, as the heat exchanger that would lead to a hot side outlet
temperature equal to the cold side inlet temperature. Similarly, the ideal heat exchanger is defined,
for the cold side, as the heat exchanger that would lead to a cold side outlet temperature equal to
the hot side inlet temperature. Wu et al. [25] introduced exergy transfer effectiveness to describe the
performance of heat exchangers and compared it with heat transfer effectiveness. g
In our study, the exergetic effectiveness on the side of hot fluid Eexh is defined as: Eexh “
.mh
“`
hh
`
Th,in
˘
´ Ta ¨ sh
`
Th,in
˘˘
´
`
hh
`
Th,ex
˘
´ Ta ¨ sh
`
Th,ex
˘˘‰
.mh
“`
hh
`
Th,in
˘
´ Ta ¨ sh
`
Th,in
˘˘
´
`
hh
`
Tc,in
˘
´ Ta ¨ sh
`
Tc,in
˘˘‰
(15) (15) with hh and sh respectively specific enthalpy and entropy of hot fluid, Th,in, Th,ex respectively inlet and
exhaust hot fluid temperatures, Tc,in inlet cold fluid temperature. This exergetic effectiveness corresponds to the ratio between the real exergetic flux transferred
from the hot fluid, i.e., the potentially recoverable mechanical power, and the mechanical power that
would have been recoverable by taking the hot fluid to the cold fluid inlet temperature. y
g
p
The exergetic effectiveness on the side of cold fluid Eexc is written: The exergetic effectiveness on the side of cold fluid Eexc is written: Eexc “
.mc
“
phc pTc,exq ´ Ta ¨ sc pTc,exqq ´
`
hc
`
Tc,in
˘
´ Ta ¨ sc
`
Tc,in
˘˘‰
.mc
“`
hc
`
Th,in
˘
´ Ta ¨ sc
`
Th,in
˘˘
´
`
hc
`
Tc,in
˘
´ Ta ¨ sc
`
Tc,in
˘˘‰
(16) (16) with hc and sc respectively specific enthalpy and entropy of cold fluid, Tc,ex exhaust cold fluid temperature. 5. Results and Discussion A comparison of the theoretical and experimental characterization of the heat exchanger supplying
the cogeneration system is conducted. The performances achieved are detailed. An analysis of the
exergetic fluxes in the micro-CHP system is presented. In the following, the ambient temperature
considered, in particular for exergetic analyses, is 293 K. 5.1. Energetic Study of the Heat Exchanger: Comparison of Theoretical and Experimental Results An experimental characterization of the heat exchanger supplying the Ericsson engine is
conducted following the procedures described in Section 3. The inlet and exhaust temperatures
of flue gas and air and the temperature of the heat exchanger wall are measured for different air
volume flow rates crossing the heat exchanger, two inlet air temperatures of 33 ˝C and 150 ˝C, and
various gas inlet temperatures. From the temperatures measured, the experimental thermal power
exchanged Pth can be estimated with the following equation: Pth “
.maircair
`
Tair,ex ´ Tair,in
˘
(17) (17) with cair the heat capacity at constant pressure and at average temperature of air. The results are given
in Table 2. Entropy 2016, 18, 154 11 of 17 Table 2. Experimental values for the heat exchanger tests. Table 2. Experimental values for the heat exchanger tests. Air Flow Rate
2.5 Nm3/h
5 Nm3/h
7.5 Nm3/h
10 Nm3/h
Inlet air temperature
33 ˝C
150 ˝C
33 ˝C
150 ˝C
33 ˝C
150 ˝C
33 ˝C
150 ˝C
Inlet flue gas temperature
980 ˝C
818 ˝C
955 ˝C
870 ˝C
954 ˝C
904 ˝C
924 ˝C
962 ˝C
Wall temperature
737 ˝C
638 ˝C
729 ˝C
666 ˝C
719 ˝C
684 ˝C
693 ˝C
717 ˝C
Outlet air temperature
232 ˝C
194 ˝C
294 ˝C
257 ˝C
325 ˝C
294 ˝C
319 ˝C
336 ˝C
Outlet flue gas temperature
936 ˝C
770 ˝C
930 ˝C
818 ˝C
914 ˝C
846 ˝C
894 ˝C
897 ˝C
Thermal power
182 W
68 W
506 W
189 W
848 W
410 W
1096 W
719 W The wall temperature Twall follows the same evolution as the inlet flue gas temperature Tgas,in
obtained during the tests for different air volume flow rates. The exhaust air temperature Tair,ex
increases with the air volume flow rate. This shows that the increase of the air velocity has a direct
impact on the global heat transfer, despite the flue gas inlet temperature variations. 5. Results and Discussion During the tests,
the exhaust air temperature is lower when the air is preheated via 6 kW electrical heating than without
preheating. This contradictory result is caused by a higher influence of the flue gas inlet temperature
on the global heat exchange compared with the inlet air temperature. The thermal power increases with the air volume flow rate and with the gas inlet temperature. In the same way as the air exhaust temperature, the thermal power exchanged is lower for the tests
when the air inlet is preheated, but the inlet flue gas temperature is lower, showing the higher impact
on the heat transfer of the inlet flue gas temperature compared with the inlet air temperature. The experimental convective heat transfer coefficient between the heat exchanger wall and the air
flow hair,cv can be defined as: hair,cv “ Pth{
`
Aair
`
Twall ´ Tair
˘˘
(18) (18) with Aair the heat exchanger wall surface area on the side of air (reference surface assumed), Twall the
temperature of heat exchanger wall, Tair the average air temperature in the heat exchanger. In the same
way, the experimental convective and radiative heat transfer coefficient between the heat exchanger
wall and the flue gas hgas,cv-R is written: with Aair the heat exchanger wall surface area on the side of air (reference surface assumed), Twall the
temperature of heat exchanger wall, Tair the average air temperature in the heat exchanger. In the same
way, the experimental convective and radiative heat transfer coefficient between the heat exchanger
wall and the flue gas hgas,cv-R is written: hgas,cv´R “ Pth{
`
Aair
`
Tgas ´ Twall
˘˘
(19) (19) with Tgas the average flue gas temperature in the heat exchanger. The experimental heat transfer
coefficients of Equations (19) and (20) depending on air volume flow rate for several inlet air
temperatures are presented in Figure 9. The convective heat transfer coefficient on the side of air is
lower than the convective and radiative heat transfer coefficient on the side of flue gas, so that the
convection phenomenon between the heat exchanger wall and the air is the first limitation to the global
heat transfer in the heat exchanger. This result corroborates the conclusions of the energetic study of
the heat exchanger presented in [15]. 12 of 17 Entropy 2016, 18, 154 Figure 9. 5. Results and Discussion The thermal power is written:
,
,
,
,
log
,
,
,
,
ln
gas in
air ex
gas ex
air in
gas in
air ex
gas ex
air in
T
T
T
T
P
FUA T
FUA
T
T
T
T
(20)
P “ FUA∆Tlog “ FUA
`
Tgas,in ´ Tair,ex
˘
´
`
Tgas,ex ´ Tair,in
˘
ln
´ Tgas,in´Tair,ex
Tgas,ex´Tair,in
¯
(20) (20)
(20) It is possible to introduce the average temperatures:
It is possible to introduce the average temperatures: It is possible to introduce the average temperatures:
It is possible to introduce the average temperatures: It is possible to introduce the average temperatures:
It is possible to introduce the average temperatures:
,
,
,
,
,
,
ln
gas
air
gas
air
gas in
air ex
gas ex
air in
gas
air
gas in
air ex
T
T
T
T
T
T
T
T
P
FUA
T
T
T
T
T
T
T
T
(21)
P “ FUA
`
Tgas ` ∆Tgas,in ´
`
Tair ` ∆Tair,ex
˘˘
´
`
Tgas ´ ∆Tgas,ex ´
`
Tair ´ ∆Tair,in
˘˘
ln
ˆ
Tgas`∆Tgas,in´pTair`∆Tair,exq
Tgas´∆Tgas,ex´pTair´∆Tair,inq
˙
(21) (21)
,
,
gas
air
gas ex
air in
T
T
T
T
where the temperature drops or gains are defined as follows:
,
,
gas
gas in
gas in
T
T
T
,
where the temperature drops or gains are defined as follows: Tgas,in “ Tgas ` ∆Tgas,in, Tgas,out “
Tgas ´ ∆Tgas,ex, Tair,in “ Tair ´ ∆Tair,in, Tair,ex “ Tair ` ∆Tair,ex. This equation can be simplified:
,
,
gas
air
gas ex
air in
where the temperature drops or gains are defined as follows:
,
,
gas
gas in
gas in
T
T
T
,
where the temperature drops or gains are defined as follows: Tgas,in “ Tgas ` ∆Tgas,in, Tgas,out “
Tgas ´ ∆Tgas,ex, Tair,in “ Tair ´ ∆Tair,in, Tair,ex “ Tair ` ∆Tair,ex. 5. Results and Discussion Evolution of the convective heat transfer coefficient of air hair and of the convective and
radiative heat transfer coefficient of flue gas hgas versus air volume flow rate for air inlet temperatures
of 33 °C and 150 °C. 0
5
10
15
20
25
30
35
2
3
4
5
6
7
8
9
10
heat exchange coefficient (W/m2.K)
air volume flow rate (Nm3/h)
hgas(Tair,in=33°C)
hgas(Tair,in=150°C)
hair(Tair,in=33°C)
hair(Tair,in=150°C)
hgas (Tair,in=33°C)
hair (Tair,in=33°C)
hgas (Tair,in=150°C)
hair (Tair,in=150°C)
Figure 9. Evolution of the convective heat transfer coefficient of air hair and of the convective and
radiative heat transfer coefficient of flue gas hgas versus air volume flow rate for air inlet temperatures
of 33 ˝C and 150 ˝C. 0
5
10
15
20
25
30
35
2
3
4
5
6
7
8
9
10
heat exchange coefficient (W/m2.K)
air volume flow rate (Nm3/h)
hgas(Tair,in=33°C)
hgas(Tair,in=150°C)
hair(Tair,in=33°C)
hair(Tair,in=150°C)
hgas (Tair,in=33°C)
hair (Tair,in=33°C)
hgas (Tair,in=150°C)
hair (Tair,in=150°C) air volume flow rate (Nm3/h) Figure 9. Evolution of the convective heat transfer coefficient of air hair and of the convective and
radiative heat transfer coefficient of flue gas hgas versus air volume flow rate for air inlet temperatures
of 33 °C and 150 °C. Figure 9. Evolution of the convective heat transfer coefficient of air hair and of the convective and
radiative heat transfer coefficient of flue gas hgas versus air volume flow rate for air inlet temperatures
of 33 ˝C and 150 ˝C. It is important to know if the design of the heat exchanger leads to the possible application of
the NTU-Effectiveness method. For example, it is necessary to check that the design of the headers
does not lead to flow maldistribution that are known to decrease the effectiveness of any heat
exchanger [26]. Hence, it is necessary to estimate the global heat transfer coefficient by another
method: the log mean temperature difference. The thermal power is written:
T
T
T
T
It is important to know if the design of the heat exchanger leads to the possible application of the
NTU-Effectiveness method. For example, it is necessary to check that the design of the headers does
not lead to flow maldistribution that are known to decrease the effectiveness of any heat exchanger [26]. Hence, it is necessary to estimate the global heat transfer coefficient by another method: the log mean
temperature difference. 5. Results and Discussion This equation can be simplified: ,
,
gas
out
gas ex
T
T
,
,
,
air
air in
air in
T
T
T
,
,
,
air
air ex
air ex
T
T
T
. The correction factor depends on the Pc and Rc factors [27] (cf. Table 3).
p
c
c
[
] ( f
T bl 3 E
i
l
l
f h
P
d R
f ˘
The experimental global heat exchange coefficient can also be evaluated using the Number of
Transfer Units of Section 4.1:
/
NTU
A
U
The experimental global heat exchange coefficient can also be evaluated using the Number of
Transfer Units of Section 4.1:
The experimental global heat exchange coefficient can also be evaluated using the Number of
Transfer Units of Section 4.1:
U The experimental global heat exchange coefficient can also be evaluated using the Number of
Transfer Units of Section 4.1:
UNTU “
`
NTUc
.mair cair
˘
{Aair
(26)
The experimental global heat exchange coefficient can also be evaluated using the Number of
Transfer Units of Section 4.1:
/
c
air
air
air
NTU
NTU m
c
A
U
(26) (26)
(26) UNTU “
`
NTUc
.mair cair
˘
{Aair
(26)
/
c
air
air
air
NTU
NTU m
c
A
U
(26) The experimental global heat exchange coefficients evaluated with Equations (20) and (26),
depending on the air volume flow rate for several air inlet temperatures, are represented in Figure 10. The results are superposed, that shows the appropriateness of the NTU method to study the heat
transfer through the heat exchanger, even when a radiative heat transfer from flue gas occurs and
that justifies also the approximation used for the estimation of Uapproximate. The influence of the air
volume flow rate and of the inlet air temperature on the global heat exchange coefficient is similar as
on the thermal power exchanged (cf. Table 2). The global heat exchange coefficient is situated between
0.47 and 6.8 W/m2¨ K for the studied range of air volume flow rates. This low value is linked to the
low air volume flow rate in the heat exchanger for a large flow section, which leads to a laminar flow
of air and to a limited convective heat exchange between air and heat exchanger wall (cf. Figure 9). The experimental global heat exchange coefficients evaluated with Equations (20) and (26),
depending on the air volume flow rate for several air inlet temperatures, are represented in Figure 10. The results are superposed, that shows the appropriateness of the NTU method to study the heat
transfer through the heat exchanger, even when a radiative heat transfer from flue gas occurs and
that justifies also the approximation used for the estimation of Uapproximate. The correction factor depends on the Pc and Rc factors [27] (cf. Table 3).
p
c
c
[
] ( f
T bl 3 E
i
l
l
f h
P
d R
f The correction factor depends on the Pc and Rc factors [27] (cf. Table 3). p
c
c
[
] ( f
bl
E
l
l
f h
P
d R
f The correction factor depends on the Pc and Rc factors [27] (cf. Table 3). p
c
c
[
] ( f
T bl 3 E
i
l
l
f h
P
d R
f The correction factor depends on the Pc and Rc factors [27] (cf. Table 3). p
c
c
[
] ( f
T bl 3 E
i
l
l
f h
P
d R
f Table 3. Experimental values of the Pc and Rc factors. p
c
c
2 5 Nm3/h
5 Nm3/h
7 5 Nm3/h Table 3. Experimental values of the Pc and Rc factors. Air Flow Rate
2.5 Nm3/h
5 Nm3/h
7.5 Nm3/h
10 Nm3/h
Pc
0.21
0.066
0.28
0.14
0.32
0.19
0.32
0.23
Rc
0.22
1.1
0.096
0.49
0.14
0.40
0.10
0.35
Air Flow Rate
2.5 Nm3/h
5 Nm3/h
7.5 Nm3/h
10 Nm3/h
cP
0.21
0.066
0.28
0.14
0.32
0.19
0.32
0.23
c
R
0.22
1.1
0.096
0.49
0.14
0.40
0.10
0.35
can be concluded that in all experimental conditions, the correction factor is unity. Hen It can be concluded that in all experimental conditions, the correction factor is unity. Hence, the thermal power transmitted across the heat exchanger can be approximated by
Pth “ UapproximateAair
`
Tgas ´ Tair
˘
. Inserting Equation (18) in this equation leads to an estimation
of the global heat exchange coefficient:
thermal
power
transmitted
across
the
heat
exchanger
can
be
approximated
by
gas
air
th
approximate
air
P
U
A
T
T
. Inserting Equation (18) in this equation leads to an estimation of the
global heat exchange coefficient:
i
i
i i
m c
T
T
Uapproximate “
.maircair
`
Tair,ex ´ Tair,in
˘
Aair
`
Tgas ´ Tair
˘
(25)
,
,
approximate
gas
air
air
U
A
T
T
(25)
h
h
ff
l
b
l
d
h
b
f (25)
(
) The experimental global heat exchange coefficient can also be evaluated using the Number of
Transfer Units of Section 4.1:
`
. 5. Results and Discussion This equation can be simplified:
,
,
,
,
,
,
,
,
gas in
gas ex
air ex
air in
gas in
gas ex
air ex
air in
T
T
T
T
T
T
T
T
P
FUA
FUA
T
T
T
T
T
T
T
T
(22)
P “ FUA
∆Tgas,in`∆Tgas,ex´p∆Tair,ex`∆Tair,inq
ln
˜
1`
∆Tgas,in`∆Tgas,ex´p∆Tair,ex`∆Tair,inq
Tgas´Tair`∆Tair,in´∆Tgas,ex
¸ “ FUA
∆Tgas,in`∆Tgas,ex´p∆Tair,ex`∆Tair,inq
ln
˜
1`pTgas,in´Tgas,exq´pTair,ex´Tair,inq
Tgas´Tair`∆Tair,in´∆Tgas,ex
¸
(22) (22)
,
,
,
,
,
,
,
,
,
,
ln 1
ln 1
gas in
gas ex
air ex
air in
gas in
gas ex
air ex
air in
gas
air
gas
air
air in
gas ex
T
T
T
T
T
T
T
,
,
air in
gas ex
T
(22)
If h
diff
( T
T
)
d ( T
T
)
h
ll
h
h
If the temperature differences (Tair,ex ´ Tair,in) and (Tgas,in ´ Tgas,ex) are much smaller than the
difference of the mean temperatures (Tgas ´ Tair), it can be assumed that:
,
,
,
,
,
,
,
,
,
,
ln 1
ln 1
gas in
gas ex
air ex
air in
gas in
gas ex
air ex
air in
gas
air
gas
air
air in
gas ex
T
T
T
T
T
T
T
,
,
air in
gas ex
T
(22)
If h
d ff
d
h
ll
h
h
If the temperature differences (Tair,ex ´ Tair,in) and (Tgas,in ´ Tgas,ex) are much smaller than the
difference of the mean temperatures (Tgas ´ Tair), it can be assumed that: If the temperature differences (
,
,
air ex
air in
T
T
) and (
,
,
gas in
gas ex
T
T
) are much smaller than the
difference of the mean temperatures (
gas
air
T
T
), it can be assumed that:
T
T
T
T
T
T
T
T
ln
˜
1 ` ∆Tgas,in ` ∆Tgas,ex ´
`
∆Tair,ex ` ∆Tair,in
˘
Tgas ´ Tair ` ∆Tair,in ´ ∆Tgas,ex
¸
« ∆Tgas,in ` ∆Tgas,ex ´
`
∆Tair,ex ` ∆Tair,in
˘
Tgas ´ Tair ` ∆Tair,in ´ ∆Tgas,ex
(23) (23) ,
ln 1
gas in
g
gas
ai
T
T
and Equation (22) reduces to: ) reduces to:
P “ FUA
`
Tgas ´ Tair ` ∆Tair,in ´ ∆Tgas,ex
˘
« FUA
`
Tgas ´ Tair
˘
(24) ) reduces to:
P “ FUA
`
Tgas ´ Tair ` ∆Tair,in ´ ∆Tgas,ex
˘
« FUA
`
Tgas ´ Tair
˘ (24) 13 of 17
13 of 17 Entropy 2016, 18, 154
Entropy 2016, 18, 154 The correction factor depends on the Pc and Rc factors [27] (cf. Table 3).
p
c
c
[
] ( f
T bl 3 E
i
l
l
f h
P
d R
f The influence of the air
volume flow rate and of the inlet air temperature on the global heat exchange coefficient is similar as
on the thermal power exchanged (cf. Table 2). The global heat exchange coefficient is situated between
0.47 and 6.8 W/m2·K for the studied range of air volume flow rates. This low value is linked to the
low air volume flow rate in the heat exchanger for a large flow section, which leads to a laminar flow
of air and to a limited convective heat exchange between air and heat exchanger wall (cf. Figure 9). Figure 10. Comparison of the experimental global heat exchange coefficient evaluated from First Law
of Thermodynamics (Uapproximate) and from the NTU method (UNTU) for air inlet temperatures of 33 °C
and 150 °C. 0
1
2
3
4
5
6
7
8
2
3
4
5
6
7
8
9
10
global heat exchange coefficient (W/m2.K)
air volume flow rate (Nm3/h)
Série1
Série3
Tair,in = 33 °C
Tair,in = 150 °C
Uapproximate
UNTU
Figure 10. Comparison of the experimental global heat exchange coefficient evaluated from First Law
of Thermodynamics (Uapproximate) and from the NTU method (UNTU) for air inlet temperatures of 33 ˝C
and 150 ˝C. 4
5
6
7
8
9
1
air volume flow rate (Nm3/h) Figure 10. Comparison of the experimental global heat exchange coefficient evaluated from First Law
of Thermodynamics (Uapproximate) and from the NTU method (UNTU) for air inlet temperatures of 33 °C
and 150 °C. Figure 10. Comparison of the experimental global heat exchange coefficient evaluated from First Law
of Thermodynamics (Uapproximate) and from the NTU method (UNTU) for air inlet temperatures of 33 ˝C
and 150 ˝C. Figure 10. Comparison of the experimental global heat exchange coefficient evaluated from First Law
of Thermodynamics (Uapproximate) and from the NTU method (UNTU) for air inlet temperatures of 33 °C
and 150 °C. Figure 10. Comparison of the experimental global heat exchange coefficient evaluated from First Law
of Thermodynamics (Uapproximate) and from the NTU method (UNTU) for air inlet temperatures of 33 ˝C
and 150 ˝C. Entropy 2016, 18, 154 14 of 17 5.2. Performances of the Heat Exchanger Integrated to The Solid Biomass-Fuelled Micro-CHP System 5.2. Performances of the Heat Exchanger Integrated to The Solid Biomass-Fuelled Micro-CHP System 5.2. The correction factor depends on the Pc and Rc factors [27] (cf. Table 3).
p
c
c
[
] ( f
T bl 3 E
i
l
l
f h
P
d R
f Performances of the Heat Exchanger Integrated to The Solid Biomass-Fuelled Micro-CHP System The experimental performances of the heat exchanger are detailed in Table 4. The heat exchanger
hot fluid effectiveness and cold fluid effectiveness are low and may be enhanced by using pressurized
air as considered in the theoretical energetic study [15], or by increasing the heat exchange surface area. Table 4. Experimental performances of the heat exchanger for air volume flow rate of 10 Nm3/h and
air inlet temperature of 150 ˝C. Table 4. Experimental performances of the heat exchanger for air volume flow rate of 10 Nm3/h and
air inlet temperature of 150 ˝C. Parameter
Value
Tair,in
150 ˝C
Tair,ex
336 ˝C
Tgas,in
962 ˝C
Tgas,ex
897 ˝C
Rc
0.354
Rh
2.826
NTUair (on the side of air)
0.271
NTUgas (on the side of flue gas)
0.096
Effectiveness Eair
22.7%
Effectiveness Egas
8.1%
Exchange surface area on the side of air Aair
0.22 m2
Thermal power transferred
719 W
Exergetic effectiveness Eexc (on the side of air)
13.8%
Exergetic effectiveness Eexh (on the side of flue gas)
10.5%
Exergetic efficiency
56.5% 5.3. Exergetic Flux and Exergy Destruction in the Micro-CHP System 5.3. Exergetic Flux and Exergy Destruction in the Micro-CHP System 6. Conclusions
6. Conclusions To study a heat exchanger intended for the heat supply of an Ericsson engine associated to a
solid biomass-fuelled micro-CHP system, an experimental test bench is implemented, with the
measurement of air mass flow rate and temperatures of inlet and exhaust air, flue gas and wall. The
objective of the paper is firstly to validate an energetic model of the heat transfer crossing the heat
exchanger inserted in the micro-CHP unit with experimental results, and secondly to present an
exergy analysis of the heat exchanger. The energetic model is based on a NTU method and the exergy
fluxes associated to the heat exchanger are modelled, including the estimation of the specific chemical
exergy of solid biomass fuel. The exergy and energy analysis are complementary. To optimize the
micro-CHP system, the exergy analysis is necessary to calculate the destructed exergetic fluxes for
each component, as the heat exchanger located inside the boiler. To complete the estimation of the
heat exchanger exergetic efficiciency which is the ratio between produced exergy and resource
exergy, a new exergetic parameter is introduced: the exergetic effectiveness, allowing a comparison
between the real and the ideal heat exchanger considering the Second Law of Thermodynamics. The
results of the experimental tests are compared with the energetic study and corroborate the
theoretical results previously obtained [15]. The convection phenomenon between air and heat
exchanger wall is the first limit to the global heat transfer. The comparison of experimental global
heat transfer coefficients evaluated with an energy balance and with the NTU model shows that the
NTU method is adapted to model heat exchangers including radiation from flue gas. Concerning the
energy and exergy analysis of the heat exchanger, compared with the air side, the effectiveness and
exergetic effectiveness on the flue gas side are lower because of the low energy quantity transferred
to air regarding the high energy quantity of flue gas. They may be enhanced by using pressurized air
as considered in the theoretical energetic study [15], or by increasing the heat exchange surface area. The evaluation of the performances of the heat exchanger is more objective when using exergetic
performances than the usual effectiveness from NTU method (performance with energetic aspect)
because the entropy generated when the temperatures of the fluids change is considered in the
calculations. 6. Conclusions
6. Conclusions The evaluation of the exergetic fluxes and the exergy destruction in the micro-CHP
system shows that the combustion of solid biomass presents the highest exergy destruction. The heat
exchanger supplying the Ericsson engine has a low impact on the total exergy destruction of the
system because of the low heat exchanged A fraction of the exergy destruction linked to heat
To study a heat exchanger intended for the heat supply of an Ericsson engine associated to a solid
biomass-fuelled micro-CHP system, an experimental test bench is implemented, with the measurement
of air mass flow rate and temperatures of inlet and exhaust air, flue gas and wall. The objective of the
paper is firstly to validate an energetic model of the heat transfer crossing the heat exchanger inserted
in the micro-CHP unit with experimental results, and secondly to present an exergy analysis of the
heat exchanger. The energetic model is based on a NTU method and the exergy fluxes associated to the
heat exchanger are modelled, including the estimation of the specific chemical exergy of solid biomass
fuel. The exergy and energy analysis are complementary. To optimize the micro-CHP system, the
exergy analysis is necessary to calculate the destructed exergetic fluxes for each component, as the
heat exchanger located inside the boiler. To complete the estimation of the heat exchanger exergetic
efficiciency which is the ratio between produced exergy and resource exergy, a new exergetic parameter
is introduced: the exergetic effectiveness, allowing a comparison between the real and the ideal heat
exchanger considering the Second Law of Thermodynamics. The results of the experimental tests are
compared with the energetic study and corroborate the theoretical results previously obtained [15]. The convection phenomenon between air and heat exchanger wall is the first limit to the global heat
transfer. The comparison of experimental global heat transfer coefficients evaluated with an energy
balance and with the NTU model shows that the NTU method is adapted to model heat exchangers
including radiation from flue gas. Concerning the energy and exergy analysis of the heat exchanger,
compared with the air side, the effectiveness and exergetic effectiveness on the flue gas side are lower
because of the low energy quantity transferred to air regarding the high energy quantity of flue gas. They may be enhanced by using pressurized air as considered in the theoretical energetic study [15],
or by increasing the heat exchange surface area. 5.3. Exergetic Flux and Exergy Destruction in the Micro-CHP System The exergetic efficiency of the global solid biomass-fuelled micro-CHP system is written: The exergetic efficiency of the global solid biomass-fuelled micro-CHP system is written: ηex,µCHP “
. Ex
. W
coge `
. Exair,ex
coge `
. Exgas,ex
Hw
`
. Exwater,ex
Hw
. Exair,in
comb `
. Exbiomass
comb
`
. Exair,in
coge `
. Exwater,in
Hw
(27) (27) This definition corresponds to a maximum theoretical exergetic efficiency and reaches the value
of 25.2%, with the following main data of the system: ‚
For the gas-water heat exchanger (Hw): mass flow rate of water = 0.5 kg/s, mass flow rate of
gas = 0.022 kg/s, water inlet temperature = 60 ˝C, water outlet temperature = 80 ˝C, gas outlet
temperature = 180 ˝C. ‚
For the gas-water heat exchanger (Hw): mass flow rate of water = 0.5 kg/s, mass flow rate of
gas = 0.022 kg/s, water inlet temperature = 60 ˝C, water outlet temperature = 80 ˝C, gas outlet
temperature = 180 ˝C. ‚
For the combustion chamber (comb): mass flow rate of pellet = 0.002 kg/s, mass flow rate of
air = 0.020 kg/s, air inlet temperature = 20 ˝C, outlet gas temperature = 800 ˝C. ‚
For the Ericsson engine (coge): mass flow rate of air = 0.0019 kg/s, air inlet temperature = 20 ˝C,
air outlet temperature = 198 ˝C. The Sankey diagram of the exergetic fluxes associated to the micro-CHP system is presented in
Figure 11. The arrows associated to exergy destruction represent both recoverable and unrecoverable
exergetic fluxes. The highest exergy destruction is linked to solid biomass combustion. The exergy
destruction in the heat exchanger supplying the Ericsson engine is low, when compared to the exergy
destruction of the water heat exchanger, mainly due to the difference in thermal power exchanged in
both heat exchangers. A fraction of the exergy destruction, linked to heat transfers at the micro-CHP
components walls, may be avoided by insulating the components that might enhance the exergetic
performances of the system. However, from the present study, it is not possible to estimate the fraction
of unrecoverable exergy (associated to entropy generation) in the exergy destruction. 15 of 17
15 of 17 Entropy 2016, 18, 154
Entropy 2016 18 154 ntropy 2016, 18, 154
15 of
ntropy 2016, 18, 154
15 o
Figure 11. Sankey diagram of the exergetic fluxes in the solid biomass-fuelled micro-CHP system. Figure 11. 5.3. Exergetic Flux and Exergy Destruction in the Micro-CHP System Sankey diagram of the exergetic fluxes in the solid biomass-fuelled micro-CHP system. Figure 11. Sankey diagram of the exergetic fluxes in the solid biomass-fuelled micro-CHP system. Figure 11. Sankey diagram of the exergetic fluxes in the solid biomass-fuelled micro-CHP system. 6. Conclusions
6. Conclusions The evaluation of the performances of the heat
exchanger is more objective when using exergetic performances than the usual effectiveness from NTU
method (performance with energetic aspect) because the entropy generated when the temperatures
of the fluids change is considered in the calculations. The evaluation of the exergetic fluxes and the
exergy destruction in the micro-CHP system shows that the combustion of solid biomass presents
the highest exergy destruction. The heat exchanger supplying the Ericsson engine has a low impact
on the total exergy destruction of the system, because of the low heat exchanged. A fraction of the 16 of 17 Entropy 2016, 18, 154 exergy destruction, linked to heat transfers at the micro-CHP components walls, may be avoided by
insulating the components that might enhance the exergetic performances of the system. exergy destruction, linked to heat transfers at the micro-CHP components walls, may be avoided by
insulating the components that might enhance the exergetic performances of the system. Acknowledgments: This work has been performed in the framework of the regional project Sylwatt, carried out in
partnership with the laboratories LAMIH (Valenciennes, France), PC2A (Lille, France), CCM (Dunkerque, France). A financial support has been provided by the Région Nord-Pas-de-Calais, by the French National Association
of Research and Technology ANRT (doctoral scholarship) and by the company Enerbiom. The authors are very
grateful to all the persons involved in the setting up of the heat exchanger test bench and in the experimental
tests. They thank in particular Sébastien Coupeaux and Annick Many (UVHC), Marc Lippert and Jesse Schiffler
(LAMIH), Federico Spoldi (LAMIH). Author Contributions: This study synthetizes the research work investigated by Marie Creyx, Eric Delacourt,
Céline Morin, Sylvain Lalot and Bernard Desmet. The energetic and exergetic models were developed by
Marie Creyx. Experimental characterization was performed by Marie Creyx and Eric Delacourt. Sylvain Lalot
developed the NTU method. All authors analyzed the results, wrote and revised the paper. All authors have read
and approved the final manuscript. Conflicts of Interest: The authors declare no conflict of interest. Conflicts of Interest: The authors declare no conflict of interest. References 1. Crema, L.; Alberti, F.; Bertaso, A.; Bozzoli, A. Development of a pellet boiler with Stirling engine for m-CHP
domestic application. Energy Sustain. Soc. 2011, 1, 1–11. [CrossRef] 1. Crema, L.; Alberti, F.; Bertaso, A.; Bozzoli, A. Development of a pellet boiler with Stirling engine for m-CHP
domestic application. Energy Sustain. Soc. 2011, 1, 1–11. [CrossRef] 2. Thiers, S.; Aoun, B.; Peuportier, B. Experimental characterization, modeling and simulation of a wood pellet
micro-combined heat and power unit used as a heat source for a residential building. Energy Build. 2010, 42,
896–903. [CrossRef] 2. Thiers, S.; Aoun, B.; Peuportier, B. Experimental characterization, modeling and simulation of a wood pellet
micro-combined heat and power unit used as a heat source for a residential building. Energy Build. 2010, 42,
896–903. [CrossRef] 3. Biedermann, F.; Carlsen, H.; Schöch, M.; Obernberger, I. Operating experiences with a small-scale CHP pilot
plant based on a 35 kWel hermetic four cylinder Stirling engine for biomass fuels. In Proceedings of the 11th
International Stirling Engine Conference, Rome, Italy, 19–21 November 2003; pp. 19–21. 3. Biedermann, F.; Carlsen, H.; Schöch, M.; Obernberger, I. Operating experiences with a small-scale CHP pilot
plant based on a 35 kWel hermetic four cylinder Stirling engine for biomass fuels. In Proceedings of the 11th
International Stirling Engine Conference, Rome, Italy, 19–21 November 2003; pp. 19–21. 4. Podesser, E. Electricity production in rural villages with a biomass Stirling engine. Renew. Energy 1999, 16,
1049–1052. [CrossRef] 4. Podesser, E. Electricity production in rural villages with a biomass Stirling engine. Renew. Energy 1999, 16,
1049–1052. [CrossRef] 5. Qiu, K.; Hayden, A.C.S. Integrated thermoelectric and organic Rankine cycles for micro-CHP systems. Appl. Energy 2012, 97, 667–672. [CrossRef] 5. Qiu, K.; Hayden, A.C.S. Integrated thermoelectric and organic Rankine cycles for micro-CHP systems. Appl. Energy 2012, 97, 667–672. [CrossRef] 6. Alanne, K.; Laukkanen, T.; Saari, K.; Jokisalo, J. Analysis of a wooden pellet-fueled domestic thermoelectric
cogeneration system. Appl. Therm. Eng. 2014, 63, 1–10. [CrossRef] 6. Alanne, K.; Laukkanen, T.; Saari, K.; Jokisalo, J. Analysis of a wooden pellet-fueled domestic thermoelectric
cogeneration system. Appl. Therm. Eng. 2014, 63, 1–10. [CrossRef] 7. Creyx, M.; Delacourt, E.; Morin, C.; Desmet, B.; Peultier, P. Energetic optimization of the performances of
a hot air engine for micro-CHP systems working with a Joule or an Ericsson cycle. Energy 2013, 49, 229–239. [CrossRef] 7. Creyx, M.; Delacourt, E.; Morin, C.; Desmet, B.; Peultier, P. References Energetic optimization of the performances of
a hot air engine for micro-CHP systems working with a Joule or an Ericsson cycle. Energy 2013, 49, 229–239. [CrossRef] 8. Bell, M.A.; Partridge, T. Thermodynamic design of a reciprocating Joule cycle engine. Proc. Inst. Me
Part A 2003, 217, 239–246. [CrossRef] -Attab, K.A.; Zainal, Z.A. Performance of high-temperature heat exchangers in biomass fuel powered
ternally fired gas turbine systems. Renew. Energy 2010, 35, 913–920. [CrossRef] 10. Gaderer, M.; Gallmetzer, G.; Spliethoff, H. Biomass fired hot air gas turbine with fluidized bed combustion. Appl. Therm. Eng. 2010, 30, 1594–1600. [CrossRef] 11. McDonald, C.F. Recuperator considerations for future higher efficiency microturbines. Appl. Therm. Eng. 2003, 23, 1463–1487. [CrossRef] 12. El-Ehwany, A.A.; Hennes, G.M.; Eid, E.I.; El-Kenany, E. Experimental investigation of the performance
of an elbow-bend type heat exchanger with a water tube bank used as a heater or cooler in alpha-type
Stirling machines. Renew. Energy 2011, 36, 488–497. [CrossRef] 13. Carlsen, H.; Marinitsch, G.; Schöch, M.; Obernberger, I. Development of a hot heat exchanger and a cleaning
system for a 35 kW hermetic four cylinder Stirling engine for solid biomass fuels. In Proceedings of the 12th
International Stirling Engine Conference and Technology Exhibition, Durham, UK, 7–9 September 2005. 13. Carlsen, H.; Marinitsch, G.; Schöch, M.; Obernberger, I. Development of a hot heat exchanger and a cleaning
system for a 35 kW hermetic four cylinder Stirling engine for solid biomass fuels. In Proceedings of the 12th
International Stirling Engine Conference and Technology Exhibition, Durham, UK, 7–9 September 2005. 14. Bonnet, S.; Alaphilippe, M.; Stouffs, P. Energy, exergy and cost analysis of a micro-cogeneration system based
on an Ericsson engine. Int. J. Therm. Sci. 2005, 44, 1161–1168. [CrossRef] 14. Bonnet, S.; Alaphilippe, M.; Stouffs, P. Energy, exergy and cost analysis of a micro-cogeneration system based
on an Ericsson engine. Int. J. Therm. Sci. 2005, 44, 1161–1168. [CrossRef] 15. Creyx, M.; Delacourt, E.; Morin, C.; Desmet, B.; Peultier, P. Modelling of the heat transfers in a flue gas-air
heat exchanger implanted in a solid biomass fuelled micro-combined heat and electrical power system. In Proceedings of the Eurotherm Seminar No 96, Brussels, Belgium, 17–18 September 2013. 15. Creyx, M.; Delacourt, E.; Morin, C.; Desmet, B.; Peultier, P. Modelling of the heat transfers in a flue gas-air
heat exchanger implanted in a solid biomass fuelled micro-combined heat and electrical power system. © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC-BY) license (http://creativecommons.org/licenses/by/4.0/). References In Proceedings of the Eurotherm Seminar No 96, Brussels, Belgium, 17–18 September 2013. Entropy 2016, 18, 154 17 of 17 16. Lalot, S. The NTU-Effectiveness Method; Nova Science Publishers: New York, NY, USA, 2011. 17. Tessé, L.; Lamet, J.-M. Radiative transfer modeling developed at Onera for numerical simulations of
reactive flows. Aerosp. Lab 2011, 2, 1–19. 8. Rivière, P.; Soufiani, A. Updated band model parameters for H2O, CO2, CH4 and CO radiatio
high temperature. Int. J. Heat Mass Transf. 2012, 55, 3349–3358. [CrossRef] 19. Chase, M.W., Jr., Ed.; NIST-JANAF Themochemical Tables, Fourth Edition. J. Phys. Chem. Ref. Data Monogr. 1998, 9, 1–1951. 20. Song, G.; Xiao, J.; Zhao, H.; Shen, L. A unified correlation for estimating specific chemical exergy of solid
and liquid fuels. Energy 2012, 40, 164–173. [CrossRef] 21. Al-Sulaiman, F.A.; Hamdullahpur, F.; Dincer, I. Greenhouse gas emission and exergy assessments of
an integrated organic Rankine cycle with a biomass combustor for combined cooling, heating and
power production. Appl. Therm. Eng. 2011, 31, 439–446. [CrossRef] 22. Szargut, J. Exergy Method: Technical and Ecological Applications; WIT Press: Southampton, UK; Boston, MA,
USA, 2005. 23. Szargut, J.; Morris, D.R.; Steward, F.R. Exergy Analysis of Thermal, Chemical, and Metallurgical Processes;
Hemisphere: New York, NY, USA, 1988. 24. Shieh, J.H.; Fan, L.T. Estimation of Energy (Enthalpy) and Exergy (Availability) Contents in Stru
Complicated Materials. Energy Sources 1982, 6, 1–46. [CrossRef] 25. Wu, S.Y.; Yuan, X.F.; Li, Y.R.; Xiao, L. Exergy transfer effectiveness on heat exchanger for finite pressure drop. Energy 2007, 32, 2110–2120. [CrossRef] 26. Lalot, S.; Florent, P.; Lang, S.K.; Bergles, A.E. Flow maldistribution in heat exchangers. Appl. Therm. Eng. 1999, 19, 847–863. [CrossRef] 27. Bergman, T.L.; Lavine, A.S.; Incropera, F.P.; DeWitt, D.P. Fundamentals of Heat and Mass Transfer; Wiley:
New York, NY, USA, 2011. © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
|
https://openalex.org/W2008349996
|
https://www.scielo.br/j/isz/a/mTtMPDSLV3bZ8YpcqQRPqvQ/?lang=en&format=pdf
|
English
| null |
On the Neotropical genus Cybaeodamus (Araneae, Zodariidae, Storeninae)
|
Iheringia. Série zoologia/Iheringia. Série Zoologia
| 2,009
|
cc-by
| 7,044
|
PALAVRAS-CHAVE. Aranhas, Zodariidae, Região Neotropical, taxonomia. In the revision of the genera of Zodariidae, JOCQUÉ
(1991) divided the family in six subfamilies that are:
Cyriocteinae
Jocqué,
Lachesaninae
Jocqué,
Storenomorphinae Simon, Cydrelinae Simon, Zodariinae
Simon, and Storeninae Simon. The genus Cybaeodamus
Mello-Leitão, 1938 belongs to this last subfamily. This
subfamily is diagnosed by JOCQUÉ (1991) by its
representatives having “metatarsal ventral hair tufts with
hollow chisel-shaped hairs, hinged hairs are few, and
restricted to the dorsal side of the leg segments, except
in Cybaeodamus where they are numerous”. In this paper all know species of the genus are
revised based on the examination of the types and four
new species are described, two from setentrional region
of Brazil and two from meridional South America,
comprising the south of Brazil, Uruguay and north of
Argentina and new illustrations of the species are
presented. On the Neotropical genus Cybaeodamus (Araneae,
Zodariidae, Storeninae) Arno A. Lise1, Ricardo Ott2 & Everton N. L. Rodrigues3 1. Laboratório de Aracnologia, Museu de Ciências e Tecnologia, Pontifícia Universidade Católica do Rio Grande do Sul, Av. Ipiranga,
6681, Prédio 40, sala 167, 90619-900 Porto Alegre, RS, Brazil. (lisearno@pucrs.br) g
(
p
)
2. Museu de Ciências Naturais, Fundação Zoobotânica do Rio Grande do Sul, Rua Dr. Salvador França, 1427, 90690-000 Porto Alegre, RS,
Brazil. (rott@fzb.rs.gov.br) 3. Programa de Pós-Graduação em Biologia Animal, Instituto de Biociências, Universidade Federal do Rio Grande do Sul, Av. Bento
Gonçalves, 9500, Bloco IV, Prédio 43435, 91501-970 Porto Alegre, RS, Brazil. (enlrodrigues@yahoo.com.br) ABSTRACT. Four new species of the spider genus Cybaeodamus Mello-Leitão, 1938 are described and illustrated: C. meridionalis sp. nov. from Brazil and Argentina, C. taim sp. nov. from Brazil, Argentina and Uruguay, C. brescoviti sp. nov. and C. tocantins sp. nov. from
setentrional region of Brazil. For the species Cybaeodamus enigmaticus (Mello-Leitão, 1939), C. lycosoides (Nicolet, 1849) and C. ornatus Mello-Leitão, 1938, new illustrations based on the examination of the types are presented. The species Cybaeodamus nigrovittatus
Mello-Leitão, 1941 which the holotype is an immature specimen, C. pallidus (Mello-Leitão, 1943) which the type was not located, C. rastellifer (Mello-Leitão, 1940) and C. scottae Mello-Leitão, 1941, both described upon juvenile specimens, all from Argentina, are
considered as species inquirendae. Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 KEYWORDS. Spiders, Zodariidae, Neotropical region, taxonomy. KEYWORDS. Spiders, Zodariidae, Neotropical region, taxonomy. RESUMO. Sobre o gênero Neotropical Cybaeodamus (Araneae, Zodariidae, Storeninae). Neste trabalho são descritas e ilustradas
quatro espécies novas do gênero Cybaeodamus Mello-Leitão, 1938: C. meridionalis sp. nov. do Brasil e Argentina, C. taim sp. nov. do
Brasil, Argentina e Uruguai, C. brescoviti sp. nov. e C. tocantins sp. nov. do Brasil setentrional. Para as espécies Cybaeodamus
enigmaticus (Mello-Leitão, 1939), C. lycosoides (Nicolet, 1849) e C. ornatus Mello-Leitão, 1938 são apresentadas novas ilustrações a
partir do exame do material-tipo. As espécies Cybaeodamus nigrovittatus Mello-Leitão, 1941 cujo holótipo é um indivíduo jovem, C. pallidus (Mello-Leitão, 1943) da qual o tipo não foi localizado, C. rastellifer (Mello-Leitão, 1940) e C. scottae Mello-Leitão, 1941,
ambas com tipos juvenis, todas da Argentina, são consideradas species inquirendae. PALAVRAS-CHAVE. Aranhas, Zodariidae, Região Neotropical, taxonomia. Cybaeodamus meridionalis sp. nov.
(Figs. 1-20, 68) Types. Holotype
, BRAZIL, Rio Grande do Sul:
Xangrilá, 09.XI.1986, A. A. Lise col. (MCN 16013). Paratypes: ,
Portão, 21.XI.1993, G. Pontes col. (MCTP 4619); 3 , Xangrilá,
23.XI.1985, A. A. Lise col. (MCN 16074);
, 2 , Xangrilá,
17.I.1987, A. A. Lise col. (MCN 16645);
, Xangrilá, 21.I.1989,
A. A. Lise col. (MCN 18141); , Viamão (Parque Estadual de Itapuã),
17.XII.2003, A. C. K. Ferreira col. (MACN 10755); , Viamão,
10.V.1990, L. A. Santos col. (MCTP 1274);
, Porto Alegre,
22.I.1989, A. A. Lise col. (MCN 18142); , Porto Alegre (Jardim
Botânico), 20.XI.1999, A. B. Bonaldo col. (MCN 31618). Distrito
Federal:
, Brasília (Área da Marinha), 29-31.X.1999, G. G. Montingelli col. (IBSP 26543). Mato Grosso do Sul: , Corumbá
(Passo da Lontra, Capões), VII.1999, A. D. Brescovit col. (IBSP
23934). Etymology. The name in apposition reffers to the
South American distribution. Diagnosis. Males of C. meridionalis sp. nov. differ
from other species of the genus by the presence of a
translucent lamella on the base of the tegulum and by the
larger distance between the apex of the tegular distal
apophysis and the median tegular apophysis (Figs. 2-5,
8). Females differ from other species of the genus by the
presence of a patch of serrate setae on the ventral face of
abdomen (Figs. 13, 14) and by the shorter length of the
copulatory ducts (Figs. 7, 10). g
Variation. Males (15). Total length: 8.00-10.00;
carapace: 4.10-5.20; femur I: 2.80-3.20. Females (10). Total
length: 9.40-12.00; carapace: 4.50-5.80; femur I: 2.70-3.00. g
p
Distribution. Brazil (Distrito Federal, Mato Grosso,
Mato Grosso do Sul, Minas Gerais, São Paulo, Santa
Catarina and Rio Grande do Sul) and Argentina (Jujuy
and Salta). Description. Male (holotype). Total length 8.20. Carapace 4.10 long, 2.80 wide, 1.50 height. Sternum 1.80
long, 1.50 wide. Carapace oval, convex, narrower on the
anterior region (Fig. 1). Cephalic region higher. General
color dark chestnut almost black. Fovea longitudinal,
short, narrow and little deep. Radial furrows of thorax
demarcate only by the presence of double row of yellow
macrosetae. Chelicerae dark chestnut, densely covered
with short spatulate setae. Promargin of the fang furrow
with two short teeth, claw short and thick. Labium
triangular with rounded apex, chestnut. Endites triangular,
convergent (Fig. 11). Sternum reddish chestnut, convex,
densely clothed with setae. Anterior and posterior row
of eyes procurved. Anterior eye row 0.68 wide, posterior
1.00. Leg formula 4.1.3.2. MATERIAL AND METHODS The material studied are deposited in the following
institutions: FCE-AR, Facultad de Ciencias, Entomología,
Marindia, Canelones (Miguel Simó); IBSP, Instituto
Butantan, São Paulo (Antonio Domingos Brescovit); LACB,
Laboratório de Aracnologia do Instituto de Ciências
Biológicas da Universidade Federal de Minas Gerais, Belo
Horizonte (Mário De Maria); MACN, Museo Argentino de
Ciencias Naturales “Bernardino Rivadavia”, Buenos Aires
(Cristina Luisa Scioscia); MCN, Museu de Ciências Naturais
da Fundação Zoobotânica do Rio Grande do Sul, Porto
Alegre (Erica Helena Buckup); MCTP, Museu de Ciências e
Tecnologia da Pontifícia Universidade Católica do Rio Grande
do Sul, Porto Alegre (Arno Antonio Lise); MLP, Museu de
La Plata, La Plata (Luis Alberto Pereira); MNHN, Museum
National d’Histoire Naturelle, Paris (Christine Rollard) and
MZSP, Museu de Zoologia da Universidade de São Paulo,
São Paulo (Ricardo Pinto-da-Rocha). Cybaeodamus was proposed by MELLO-LEITÃO
(1938) to include C. ornatus, described from Argentina. According to JOCQUÉ (1991), the males of Cybaeodamus
“are recognized by the palps with two tegular apophysis
(distal and lateral) of which the distal one is large and
complex; females have a large epigynum with a strongly
sclerotized area on both sides; the chelicerae are densely
haired in both sexes”. Up to now Cybaeodamus comprises seven
exclusively South American species (PLATNICK, 2007): C. lycosoides (Nicolet, 1849) from Chile; C. ornatus Mello-
Leitão, 1938 from Argentina, Uruguay and Peru; C. enigmaticus (Mello-Leitão, 1939), C. nigrovittatus Mello-
Leitão, 1941, C. pallidus (Mello-Leitão, 1943), C. rastellifer (Mello-Leitão, 1940) and C. scottae Mello-
Leitão, 1941 from Argentina. Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 LISE et al. 260 brown-olivaceous, remaining articles chestnut. On the
dorsal face of patellae and tibiae there are setae disposed
on longitudinal rows, remaining articles densely
setaceous. Abdomen: 4.05 long, 2.50 wide, 2.50 height. Dorsum dark chestnut, almost black on the laterals,
permitting to see the whitish yellow bottom. Venter same
color of dorsum with scavated circular spots disposed
on two longitudinal lines. Epigastric plates yellow. Anterior margin of the epigastric furrow with a chestnut
more sclerotinized edge. Palpus with the lateral apophysis
hook shaped. Distal tegular apophysis hook shaped with
a longitudinal groove where the embolus fit. Tibial
apophysis well developed (Figs. 2, 3). Descriptions and nomenclature of the male palpus
follow JOCQUÉ (1991). The study of internal reproductive
structures of female epigynum was performed immersing
it in lactic acid or in clove oil. All measurements are in
millimeters. Cybaeodamus Mello-Leitão, 1938 Cybaeodamus MELLO-LEITÃO, 1938:93; 1941:107; ROTH, 1965:290;
LEHTINEN, 1967:226. p p y
p
(
g
, )
Female (paratype, MCN 16074). Total length 9.00. Carapace 4.25 long, 2.75 wide, 1.30 height. Sternum 1.75
long, 1.55 wide. Carapace (Fig. 12) dark chestnut, region
around fovea lighter. Chelicerae dark chestnut, densely
covered with setae on the anterior face with two small
teeth on the anterior margin of the fang furrow. Labium
with rounded apex, setaceous. Sternum chestnut,
cordiform, densely setaceous. Anterior and posterior row
of eyes procurved. Leg (Figs. 17-20) formula 4.1.2.3. Measurements: I/II/III/IV: femur 2.30/2.00/1.75/2.40;
patella 1.15/1.10/1.05/1.20; tibia 1.55/1.25/1.15/1.70;
metatarsus 1.30/1.10/1.35/2.50; tarsus 1.00/1.00/1.00/1.25. Total 7.30/6.45/6.30/9.05. All articles reddish chestnut, the
femura darker. Abdomen 5.25 long, 3.50 wide, 2.95 height,
densely covered with microsetae. Color of dorsum black
with two anterior spots, one each side forming a “V” and
some yellow spots of variable size. Laterals and venter
dark chestnut almost black with many small spots
longitudinally disposed. Spinnerets (Figs. 15, 16). Epigynum (Figs. 6, 9) with a median deep depression and
above it there are two narrow and deep excavations, one
each side where are the fecundation openings. The
spermatecae are spherical and between them there are
the fecundation ducts (Figs. 7, 10). Hyltoniella MELLO-LEITÃO, 1940:29 (type species Hyltoniella
birabeni); JOCQUÉ, 1991:49. Hyltoniella synonimized by JOCQUÉ,
1991:49. Valcheta MELLO-LEITÃO, 1940:20 (type species Valcheta rastellifera);
ROTH, 1965:291 (transferred to Zodariidae); LEHTINEN, 1967:274;
JOCQUÉ, 1991:49. Valcheta synonimized by JOCQUÉ, 1991:49. Valcheta MELLO-LEITÃO, 1940:20 (type species Valcheta rastellifera);
ROTH, 1965:291 (transferred to Zodariidae); LEHTINEN, 1967:274;
JOCQUÉ, 1991:49. Valcheta synonimized by JOCQUÉ, 1991:49. MATERIAL AND METHODS The scanning electron micrographs (SEM)
were made using a electronic microscope of the Centro
de Microscopia e Microanálises of Pontifícia
Universidade Católica do Rio Grande do Sul (PUCRS) . Other material examined. ARGENTINA. Jujuy: Banda de
Los Perales,
, , 13.XII.1950, M. Birabén col. (MLP 17503).
Salta: Anta (between del Rei and Lumbada), , 17.XI.1966, M. E.
Galiano col. (MACN 10722). BRAZIL. Mato Grosso: Chapada
dos Guimarães, 9 , 20-29.XI.2000, C. Strüssman col. (MCTP
11540). Minas Gerais: Belo Horizonte (B. Pampulha),
, , A.
Campagnani col. (LACB 1321). São Paulo: São Carlos,
,
20.VII.1982, J. Henrique col. (MCN 24930). Santa Catarina:
Florianópolis (Campeche),
, I.2000, A. D. Brescovit col. (IBSP
27553). Rio Grande do Sul: Xangrilá, 3 , 5 , 18.XI.1990-
19.II.1999, A. A. Lise col. (MCTP 1082, 2905, 2973, 3940, 9560,
10484); Canoas (Capão do Corvo), , 23.IV.1966, A. A. Lise col.
(MCN 190); Viamão (Parque Estadual de Itapuã), 23 , 8 ,
18.XI.2003-10.II.2004, A. C. K. Ferreira col. (MCTP 16418-
16440); , 10.V.1990, L. A. Santos col. (MCTP 1274); Passo do
Vigário, , 03.III.1993, R. Balestrin col. (MCTP 3031); Porto
Alegre (Morro Santana), , 05.IV.1968, A. A. Lise col. (MCN
1937); , 15.II.1989, A. D. Brescovit (MCN 18325); Porto Alegre
(Jardim Botânico),
, 20.XI.1990, A. D. Brescovit col. (MCN
20106); Porto Alegre (Jardim Botânico),
, 26.XI.1992, A. B.
Bonaldo col. (MCN 22550). Cybaeodamus meridionalis sp. nov.
(Figs. 1-20, 68) Measurements I/II/III/IV: femur
2.40/2.10/2.05/2.60; patella 1.10/1.10/1.10/1.10; tibia 1.75/
1.40/1.25/1.85; metatarsus 1.80/1.55/1.80/2.75; tarsus 1.45/
1.25/1.25/1.45. Total 8.50/7.40/7.45/9.75. Coxae and femura Other material examined. ARGENTINA. Jujuy: Banda de
Los Perales,
, , 13.XII.1950, M. Birabén col. (MLP 17503). Salta: Anta (between del Rei and Lumbada), , 17.XI.1966, M. E. Galiano col. (MACN 10722). BRAZIL. Mato Grosso: Chapada
dos Guimarães, 9 , 20-29.XI.2000, C. Strüssman col. (MCTP
11540). Minas Gerais: Belo Horizonte (B. Pampulha),
, , A. Campagnani col. (LACB 1321). São Paulo: São Carlos,
,
20.VII.1982, J. Henrique col. (MCN 24930). Santa Catarina:
Florianópolis (Campeche),
, I.2000, A. D. Brescovit col. (IBSP
27553). Rio Grande do Sul: Xangrilá, 3 , 5 , 18.XI.1990-
19.II.1999, A. A. Lise col. (MCTP 1082, 2905, 2973, 3940, 9560,
10484); Canoas (Capão do Corvo), , 23.IV.1966, A. A. Lise col. (MCN 190); Viamão (Parque Estadual de Itapuã), 23 , 8 ,
18.XI.2003-10.II.2004, A. C. K. Ferreira col. (MCTP 16418-
16440); , 10.V.1990, L. A. Santos col. (MCTP 1274); Passo do
Vigário, , 03.III.1993, R. Balestrin col. (MCTP 3031); Porto
Alegre (Morro Santana), , 05.IV.1968, A. A. Lise col. (MCN
1937); , 15.II.1989, A. D. Brescovit (MCN 18325); Porto Alegre
(Jardim Botânico),
, 20.XI.1990, A. D. Brescovit col. (MCN
20106); Porto Alegre (Jardim Botânico),
, 26.XI.1992, A. B. Bonaldo col. (MCN 22550). Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 On the Neotropical genus Cybaeodamus (Araneae, Zodariidae... 261 Figures 1-7. Cybaeodamus meridionalis sp. nov.: 1, male body, dorsal view; 2-5, male palpus: 2, ventral view; 3, retrolateral view; 4,
prolateral view; 5, dorsal view; 6, 7, female epigynum: 6, ventral view; 7, dorsal view, cleared (dta, distal tegular apophysis; e, embolus;
fd, fecundation duct; mta, median tegular apophysis; s, spermatecae; t, tegulum). Figures 1-7. Cybaeodamus meridionalis sp. nov.: 1, male body, dorsal view; 2-5, male palpus: 2, ventral view; 3, retrolateral view; 4,
prolateral view; 5, dorsal view; 6, 7, female epigynum: 6, ventral view; 7, dorsal view, cleared (dta, distal tegular apophysis; e, embolus;
fd, fecundation duct; mta, median tegular apophysis; s, spermatecae; t, tegulum). Figures 1-7. Cybaeodamus meridionalis sp. nov.: 1, male body, dorsal view; 2-5, male palpus: 2, ventral view; 3, retrolateral view; 4,
prolateral view; 5, dorsal view; 6, 7, female epigynum: 6, ventral view; 7, dorsal view, cleared (dta, distal tegular apophysis; e, embolus;
fd, fecundation duct; mta, median tegular apophysis; s, spermatecae; t, tegulum). Cybaeodamus taim sp. nov.
(Figs. 21-36, 68) Diagnosis. Males of C. taim sp. nov. differ from
the males of the other species of the genus by the
presence of an excavation on the median posterior
portion of the sternum (Fig. 25) and by the presence of
setiferous basal tubercles on the coxae (III-IV), by the
tibial apophysis of the palpus with a small hook shape
projection (Fig. 24); females differs from the ones of
the other species by the epigynum having long
copulatory ducts close to each other on the portion
near the spermatecae (Figs. 27, 28). Types. Holotype
, BRAZIL, Rio Grande do Sul: Rio
Grande (Estação Ecológica do Taim), 02.XII.1986, A. D. Brescovit
col. (MCN 16548). Paratypes: , Viamão (Itapuã), 19.I.1977, M. L. Tavares col. (MCN 5047);
, , Viamão (Parque Estadual de
Itapuã), 29.VIII.2003, A. C. K. Ferreira col. (MACN 10756); 5 ,
same locality and date of the holotype, A. D. Brescovit col. (MCN
16548);
, same locality of the holotype, 02.XII.1986, E. H. Buckup col. (MCN 16301). Description. Male (holotype). Total length 9.70. Carapace 5.50 long, 3.90 wide, 2.13 height. Sternum 2.46 Etymology. The specific name is a noun in
apposition to the genus name, taken from the type locality. Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 LISE et al. 262 glabrous. Labium fulvous, setaceous with the basal ectal
margins darker. Sternum dark fulvous with a central
anterior yellow area densely setaceous, with one deep
excavation each side, on the level of coxa III (Fig. 25). Ocular area densely covered with thick black setae. Anterior and posterior eye row procurved. Legs formula
4.1.3.2. Measurements I/II/III/IV: femur 3.75/3.60/3.25/4.10;
patella 1.70/1.60/1.60/1.70; tibia 2.63/2.05/1.75/2.50; long, 1.95 wide. Carapace suboval, narrower on the
anterior portion, cephalic region higher (Figs. 21, 26). General color light fulvous, glabrous with only one median
longitudinal row of setae that extends from the ocular
area to fovea, this longitudinal and narrow. Chelicerae
fulvous, densely setaceous (Fig. 26). Endites triangular
(Fig. 25), yellow with apical black scopula covered with
long and thick setae except on the central area which is res 8-15. Cybaeodamus meridionalis sp. nov.: 8, male palpus, ventral; 9, 10, female epigynum: 9, ventral view; 10, dorsal view
es and labium; 12, carapace and chelicerae, frontal view; 13, patch setae of the ventral face of abdomen; 14, detail of seta
le spinnerets. Figures 8-15. Cybaeodamus meridionalis sp. Cybaeodamus taim sp. nov.
(Figs. 21-36, 68) nov.: 8, male palpus, ventral; 9, 10, female epigynum: 9, ventral view; 10, dorsal view; 11
endites and labium; 12, carapace and chelicerae, frontal view; 13, patch setae of the ventral face of abdomen; 14, detail of setae; 15
female spinnerets. Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 On the Neotropical genus Cybaeodamus (Araneae, Zodariidae... 263 metatarsus 2.85/2.00/2.80/3.95; tarsus 1.75/1.60/1.50/1.85. Total 12.68/10.85/10.90/14.12. Legs light fulvous, femur
lighter on the ventral and lateral faces. Coxae III and IV
with a rhombus setiferous basal tubercle, the more
developed in coxa III (Fig. 25). Legs III and IV spiny. Abdomen (Figs. 21, 35), 4.50 long, 3.30 wide, 2.95 height. Dorsum guanine white spotted with dark reddish chestnut
(Fig. 21). Venter same color of dorsum but the guanine
spots less conspicuous. Laterals with rows of dark
chestnut spots. Ventral face with a patch of many lying
granulated setae directed backward (Fig. 35). Lateral and
dorsal faces coated with no modified setae. Spinnerets
light fulvous. Tibial apophysis of palpus with a small
hook and lateral tegular apophysis lamelar (Figs. 22-24,
29-32). Chelicerae dark fulvous very setaceous. Endites light
fulvous. Labium dark fulvous. Sternum scutiform very
convex, all covered with setae, fulvous, without the
excavations that occurs in the male. Ocular area very
setaceous. Anterior and posterior eyes row procurved. Legs formula 4.1.2.3. Measurements I/II/III/IV: femur 2.63/
2.40/2.23/2.96; patella 1.31/1.25/1.15/1.35; tibia 1.81/1.48/
1.21/1.81; metatarsus 1.64/1.58/1.86/2.80; tarsus 1.16/1.15/
1.11/1.31. Total 8.55/7.86/7.56/10.23. Legs fulvous III and
IV spiny. Abdomen 4.80 long, 3.30 wide, 2.63 height. Dorsum as on the male. Venter same color of dorsum but
the guanine spots less conspicuous. Laterals with rows
of dark chestnut spots. All faces of abdomen coated with
setae (Fig. 36). Spinnerets light fulvous. Epigynum (Figs. 27, 33) with well developed spermatecae (Figs. 28, 34). metatarsus 2.85/2.00/2.80/3.95; tarsus 1.75/1.60/1.50/1.85. Total 12.68/10.85/10.90/14.12. Legs light fulvous, femur
lighter on the ventral and lateral faces. Coxae III and IV
with a rhombus setiferous basal tubercle, the more
developed in coxa III (Fig. 25). Legs III and IV spiny. Abdomen (Figs. 21, 35), 4.50 long, 3.30 wide, 2.95 height. Dorsum guanine white spotted with dark reddish chestnut
(Fig. 21). Venter same color of dorsum but the guanine
spots less conspicuous. Laterals with rows of dark
chestnut spots. Ventral face with a patch of many lying
granulated setae directed backward (Fig. 35). Lateral and
dorsal faces coated with no modified setae. Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 Cybaeodamus taim sp. nov.
(Figs. 21-36, 68) Spinnerets
light fulvous. Tibial apophysis of palpus with a small
hook and lateral tegular apophysis lamelar (Figs. 22-24,
29-32). p
p
g
Variation. Males (5). Total length: 9.50-10.80;
carapace: 5.00-6.00; femur I: 3.50-4.15. p
p
g
Variation. Males (5). Total length: 9.50-10.80;
carapace: 5.00-6.00; femur I: 3.50-4.15. Female (paratype, MCN 5047). Total lenght 8.60. Carapace 8.60 long, 2.80 wide, 1.25 height. Sternum 2.14
long, 1.58 wide. Carapace suboval narrower anteriorly. General color light fulvous, glabrous with a single
longitudinal median row of setae that extend from the
ocular area to the fovea, this longitudinal and narrow. p
Distribution. South of Brazil (Rio Grande do Sul),
Uruguay (Canelones) and Argentina (Misiones,
Mendoza, Río Negro, Neuquén, Chubut and Santa
Cruz). Figures 16-20. Cybaeodamus meridionalis sp. nov., female: 16, anterior lateral spinneret, detail; 17-20, leg I: 17, tarsus; 18, tarsus,
detail; 19, tarsal organ; 20, trichobothrium on metatarsus. Figures 16-20. Cybaeodamus meridionalis sp. nov., female: 16, anterior lateral spinneret, detail; 17-20, leg I: 17, tarsus; 18, tarsus,
detail; 19, tarsal organ; 20, trichobothrium on metatarsus. Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 LISE et al. 264 264 Figures 21-28. Cybaeodamus taim sp. nov.: 21, male body; 22-24, male palpus: 22, ventral view; 23, prolateral view; 24, retrolateral
view; 25, cephalotorax, ventral view; 26, carapace and chelicerae; 27, 28, female epigynum: 27, ventral view; 28, dorsal view, cleared. Figures 21-28. Cybaeodamus taim sp. nov.: 21, male body; 22-24, male palpus: 22, ventral view; 23, prolateral view; 24, retrolateral
view; 25, cephalotorax, ventral view; 26, carapace and chelicerae; 27, 28, female epigynum: 27, ventral view; 28, dorsal view, cleared. Cybaeodamus brescoviti sp. nov.
(Figs. 37-41, 68) Other material examined. BRAZIL. Rio Grande do Sul:
Viamão (Parque Estadual de Itapuã), 82 , 18 , 29.VIII.2003-
08.X.2003, A. C. K. Ferreira col. (MCTP 16381-17062);
ARGENTINA. Misiones: Zehura (Estr. Partridge),
, 07.VII,
without collector (MACN 10754). Mendoza: Luján de Cuyo
(Puente del Inca), 3 , 23.III.1976, 16.III.1977, A. Roig col. (MACN
10752, 10753). Río Negro: 25 de Mayo (Ñe Luan), 3 , I.1975, E. Maury col. (MACN 10749). Neuquén: Bariloche, , II.1954, M. E. Galiano col. (MACN 5421). Chubut: Gastre, , XI.1976, D. Nevez col. (MACN 10750); Chucas (Parque Nacional los Alerces),
, XI.1982, M. Ramirez col. (MACN 10751). Santa Cruz: Lago
Argentino (El Calafate), , II.1963, E. Maury col. (MACN 10748);
URUGUAY. Canelones:
Marindia,
5 ,
,
02.II.1996-
02.XII.1998, Costa col. (FCE-AR 2434-2436). Types. Holotype
, BRAZIL, Maranhão: Barreirinhas
(Parque Nacional dos Lençóis Maranhenses), 13-17.X.2001, A. D. Brescovit col. (IBSP 33995). Paratypes: (IBSP 33994), 4 (IBSP
33990, 33991, 33992, 33996); 2 (MCTP 19989), same data as
holotype. Types. Holotype
, BRAZIL, Maranhão: Barreirinhas
(Parque Nacional dos Lençóis Maranhenses), 13-17.X.2001, A. D. Brescovit col. (IBSP 33995). Paratypes: (IBSP 33994), 4 (IBSP
33990, 33991, 33992, 33996); 2 (MCTP 19989), same data as
holotype. Etymology. The specific name is a noun in
apposition to the genus name and is a patronymic in
honor to the Brazilian arachnologist Dr. Antonio
Domingos Brescovit. Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 On the Neotropical genus Cybaeodamus (Araneae, Zodariidae... 265 Diagnosis. Males of C. brescoviti sp. nov. are
milar to the those of C. enigmaticus by having similar
trolateral tibial apophysis but differ by this being wider
nding into two blunt eminences and by having a short
gular lateral apophysis (Figs. 37-39). Females differs
om the ones of the other species of the genus by the
covering the fecundation openings and by the l
spheroid spermatecae (Figs. 40, 41). Description. Male (holotype). Total length 6
Carapace 3.23 long, 2.07 wide, 2.66 height. Sternum
long, 1.30 wide. Carapace yellowish fulvous, spotted
black, glabrous with a single median longitudinal ro
gures 29-36. Cybaeodamus taim sp. nov.: 29, male palpus, ventral; 30, palpus, detail; 31, tegular distal apophisis, detail; 32, ap
gular distal apophysis, detail; 33, 34, epigynum: 33, ventral view; 34, dorsal view; 35, patch on the ventral face of male abdomen
tch on the ventral face of female abdomen. Figures 29-36. Cybaeodamus taim sp. Distribution. Known only for Argentina. Distribution. Known only for Argentina. Discussion. JOCQUÉ (1991: Fig. 93) presented a
drawing of the dorsal view of the epigynum (Fig. 93) but
we believe it does not belong to C. enigmaticus. Cybaeodamus lycosoides (Nicolet, 1849)
(Figs. 48, 49, 68) Drassus lycosoides NICOLET, 1849:453 (described female). Storena lycosoides SIMON, 1889:218. Cybaeodamus lycosoides JOCQUÉ, 1991:50 (transferred from
Storena); PLATNICK, 2007. Drassus lycosoides NICOLET, 1849:453 (described female). Storena lycosoides SIMON, 1889:218. Cybaeodamus lycosoides JOCQUÉ, 1991:50 (transferred from
Storena); PLATNICK, 2007. Type. Holotype (examined), CHILE. No locality, date or
collector designation (MNHN 9954). The vial contains a label
with the following data: “Museu Paris AR9954; Storena lycosoides
Nic. (Drassus), Chili, E. Simon dét.”. g
p p y
g
Female (paratype, IBSP 33994). Total length 6.08. Carapace elliptic, high, narrowed in front, 3.23 long, 1.95
wide, 1.15 height. Sternum 1.50 long, 1.10 wide. Carapace
with a median elliptic yellow spot that extend from the
PME to the posterior margin of fovea, remaining of
carapace shadowed with black. Thoracic furrow darker. Tegument slightly setaceous with a median longitudinal
row of more conspicuous setae. Chelicerae light chestnut,
densely revested with strong black setae. Endites,
triangular, convergent, yellow revested with long strong
black setae. Labium chestnut, bristly. Sternum light
chestnut very convex, densely setaceous. Clypeus and
ocular area very setaceous. Leg formula 4,1,3,2. Measurements I/II/III/IV: femur 1.86/1.64/1.60/1.90; patella
0.92/0.86/0.84/0.90; tibia 1.34/1.10/0.80/1.30; metatarsi 1.10/
1.00/1.26/1.74; tarsi 0.70/0.64/0.76/0.98. Total 5.92/5.24/
5.26/6.82. Legs fulvous with macrosetae. Abdomen 3.75
long, 3.15 wide, 3.25 height. Color of dorsum cream
yellowish suffused with black on the median longitudinal
portion. Venter with an elongate spot on each side, on
the posterior third, same color of dorsum. Lateral face
with a longitudinal chestnut stripe and three more
diagonal, of same color. Spinnerets yellowish, bristly. Colulus formed by two tuffs of setae. Epigynum with a
cordiform median plate, spermatecae spheroid (Figs. 40, 41). Diagnosis. Spermatecae spherical, appart from each
other by more than their diameter (Figs. 48, 49). Diagnosis. Spermatecae spherical, appart from each
other by more than their diameter (Figs. 48, 49). y
(
g
Description. See NICOLET (1849). y
g
Description. See NICOLET (1849). Distribution. Know only from the type locality. Discussion. JOCQUÉ (1991), affirms that the female
holotype is a subadult specimen. The examined holotype
is an adult what can be confirmed by the here presented
figures of ventral and dorsal views of the epigynum (Figs. 48, 49). Cybaeodamus ornatus Mello-Leitão, 1938
(Figs. 50-59, 68) Cybaeodamus ornatus MELLO-LEITÃO, 1938:93, figs. 4, 5 (described
male and female in Agelenidae); MELLO-LEITÃO, 1941:119, pl. 3,
fig. 11 (female); ROTH, 1965:290 (transferred to Zodariidae);
JOCQUÉ, 1991:50, figs. 82-87. Types. Lectotype
(examined), ARGENTINA. Buenos
Aires: La Plata (Río Santiago) (MLP 14033). Paralectotype
(examined), La Plata (Río Santiago) (MLP 14034). Paratype
(not examined), La Plata (Río Santiago), Max Birabén col. (Museu
Nacional Rio de Janeiro 58045). Variation. Males (6). Total length: 5.30-6.75;
carapace: 2.69-3.23; femur I: 1.84-2.13. Diagnosis. Male palpus with short tegular distal
apophysis; lateral tegular apophysis distally pointed,
embolus with wide base (Figs. 50, 51, 54). Female can be
recognized by having the spermatecae appart from each
other more than their diameter and by the long sinuose
copulatory ducts with basal portion appart from each
other around the spermatecae diameter (Figs. 52, 53, 55-
57) and a patch of setae on the ventral face of abdomen
(Fig. 59). Males have a patch of granulose setae on the
ventral face of abdomen (Fig. 58). Distribution. Northeast of Brazil (Maranhão). Other examined material. PERU. Puno: Entre Puno y
Pancarcolla,
, , 16.X.1983, E. Maury col. (MACN 10729).
ARGENTINA. Salta: Rosario de Lerna (El Alizal), 2 , 20.I.1981,
A. Roig col. (MACN 10725). Santiago del Estero: Moreno Cybaeodamus brescoviti sp. nov.
(Figs. 37-41, 68) Buenos Aires:
Villarino (Montes de Oca),
, 24.II.1982, E. Maury col. (MACN
10746). Catamarca: Andalgalá, (Andalgalá),
, 19.X.1972,
Endergs col. (MACN 10747). Other examined material. ARGENTINA. Buenos Aires:
Villarino (Montes de Oca),
, 24.II.1982, E. Maury col. (MACN
10746). Catamarca: Andalgalá, (Andalgalá),
, 19.X.1972,
Endergs col. (MACN 10747). Cybaeodamus brescoviti sp. nov.
(Figs. 37-41, 68) nov.: 29, male palpus, ventral; 30, palpus, detail; 31, tegular distal apophisis, detail; 32, apex of
tegular distal apophysis, detail; 33, 34, epigynum: 33, ventral view; 34, dorsal view; 35, patch on the ventral face of male abdomen; 36,
patch on the ventral face of female abdomen. Diagnosis. Males of C. brescoviti sp. nov. are
similar to the those of C. enigmaticus by having similar
retrolateral tibial apophysis but differ by this being wider
ending into two blunt eminences and by having a short
tegular lateral apophysis (Figs. 37-39). Females differs
from the ones of the other species of the genus by the
presence of a median cordiform plate on the epigynum covering the fecundation openings and by the large
spheroid spermatecae (Figs. 40, 41). Description. Male (holotype). Total length 6.75. Carapace 3.23 long, 2.07 wide, 2.66 height. Sternum 1.56
long, 1.30 wide. Carapace yellowish fulvous, spotted with
black, glabrous with a single median longitudinal row of
bristles and some on the ocular area. Fovea longitudinal, Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 LISE et al. 266 narrow. Chelicerae light reddish chestnut, densely covered
with black bristles. Endites triangular, yellow, whitish on
top, mesal apical scopula black. On the median basal third
there is a glabrous area. Labium fulvous, bristly. Sternum
with dark margins, chestnut, central portion yellowish
fulvous, scutiform, laterally carved on the level of coxae,
convex, densely coated with black setae. Ocular area with
four strong black setae. Anterior and posterior eyes row
procurved, posterior more procurved. Leg formula 4.1.3.2. Measurements I/II/III/IV: femur 1.97/1.85/1.97/2.17; patella
0.84/0.80/0.80/0.93; tibia 1.60/1.30/1.03/1.35; metatarsus
1.46/1.32/1.38/1.82; tarsus 0.84/0.84/0.96/1.13. Total 6.71/
6.11/6.14/7.14. Coxae of legs I and II fulvous, femura
greenish black, the remaining of articles yellow. Coxae of
legs III and IV dark fulvous, femura with the basal third
whitish. Tarsi spiny. Abdomen 3.10 long, 2.04 wide, 2.14
height. Color of dorsum white guanine with a median
longitudinal dark chestnut spot. Venter whitish with some
spots near the epigastric furrow. Median longitudinal area
covered with curved black spatulated setae. Spinnerets
with base and top white. Palpus with a distal curved
tegular apophysis (Fig. 37). Diagnosis. Male palpus with elongated tegular
distal apophysis, lateral tegular apophysis wide and
elongated distally (Figs. 42-45) and a patch of setae on
ventral view of abdomen (Figs. 46, 47). Description. See MELLO-LEITÃO (1939). Other examined material. ARGENTINA. Cybaeodamus enigmaticus (Mello Leitão, 1939)
(Figs. 42-47, 68) Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 268 LISE et al. erected bristles. Ocular area bristly with a median
longitudinal row of macrosetae. Leg formula 4,1,2,3. Measurements I/II/III/IV: femur 1.66/1.52/1.60/1.80; patella
0.66/0.66/0.60/0.74; tibia 1.12/0.96/0.80/1.14; metatarsus
1.02/0.96/1.02/1.40; tarsus 0.78/0.76/0.78/0.96. Total 5.24/
4.86/4.80/6.04. Legs femura dark chestnut, patellae
fulvous, tibiae yellow with basal third light chestnut,
metatarsi and tarsi yellow. Legs I and II with a dorsal
basal macroseta on tibia. Legs III and IV with many spines. Coxae III with a ventral apical unconspicuous tubercle
(Fig. 61), femur with a ventral basal tubercle (Fig. 62),
coxae IV with an assemblage of tick setae on the inner
margin. Abdomen 2.23 long, 1.48 wide, 1.60 height. Color
of dorsum white, reticulated of guanine with a dark
chestnut spot from which start three pairs of dorsum
lateral stripes of same color. Venter with the epigastric
area light fulvous, the remaining yellowish with some
guanine spots. Median central area with dark chestnut
setae. Lateral with a longitudinal light chestnut stripe. Spinnerets light chestnut. Palpus with tibial apophysis
pointed on the distal portion, on retrolateral view (Fig. 65) and concave on prolateral side (Fig. 63), tegulum (Fig. 64) globose on retrolateral view (Fig. 65). Distribution. Argentina (Salta, Santiago del
Estero, Buenos Aires, La Pampa, Rio Negro, La Rioja,
San Juan and Chubut), Peru (Puno) and Uruguay
(JOCQUÉ, 1991:52). Cybaeodamus enigmaticus (Mello Leitão, 1939)
(Figs. 42-47, 68) Rubrius enigmaticus MELLO-LEITÃO, 1939:57, figs. 23-25 (described
the male in Agelenidae); ROTH, 1967:343 (transferred to
Zodariidae incertae sedis). Rubrius enigmaticus MELLO-LEITÃO, 1939:57, figs. 23-25 (described
the male in Agelenidae); ROTH, 1967:343 (transferred to
Zodariidae incertae sedis). Hyltoniella birabeni MELLO-LEITÃO, 1940:29, figs. 31,32, pl. I, fig. 3 (described the male); JOCQUÉ, 1991:50 (Syn.). g
Description. See MELLO-LEITÃO (1938). Description. See MELLO-LEITÃO (1938). Cybaeodamus enigmaticus; JOCQUÉ, 1991:53, figs. 88-93 (only
male); PLATNICK, 2007. Other examined material. PERU. Puno: Entre Puno y
Pancarcolla,
, , 16.X.1983, E. Maury col. (MACN 10729). ARGENTINA. Salta: Rosario de Lerna (El Alizal), 2 , 20.I.1981,
A. Roig col. (MACN 10725). Santiago del Estero: Moreno Type. Holotype
(examined), ARGENTINA, Camarones,
Patagonia, A. Masarey col. (Naturhistorisches Museum Basel). Female unknown (see discussion). Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 On the Neotropical genus Cybaeodamus (Araneae, Zodariidae... 267 (Mercedes, 10 km Este Amaná), , 07.VIII.1995, D. Vezzani col. (MACN 10730); Moreno (Mercedes), , I.1996, D. Vezzani col. (MACN 10727). Buenos Aires: Bahia Blanca, , II.1942, F. Morrôs
col. (MACN 10723); Bahia Blanca (Quinta La China, 25 Km Sierra
de la Ventana), 9 , 20.III.1983, E. Maury col. (MACN 10728);
Patagones (Bahia San Blas), 2 , , no date, C. Daguerre col. (MACN
10731); Tandil, , V.1967, E. Maury col. (MACN 10732); Balcarce
(Sierra la Barrosa), 2 , 19-21.VII.1974, E. Maury col. (MACN
10738); Patagones (Carmen de Patagones, Rio Azi), , II.1975, E. Maury col. (MACN 10724); Patagones (Carmen de Patagones), , IV.1971, Aquilera col. (MACN 10735). La Pampa: Liuhel Calel, 2 ,
10.XI.1969, 22.I.1985, E. Maury col. (MACN 10726, 10734). Rio
Negro: General Roca (General Roca-Balsa), , I.1962, B. Bachmann
col. (MACN 5468). La Rioja: Independencia (Patquia), , X.1968,
M. E. Galiano col. (MACN 10740); Arauco (Ascha Almogasta), 5 ,
2 , 1941, 1943, X.1947, J. C. Freye col. (MACN 2118, 2178,
10723, 10743, 10745). San Juan: Valle Fertil, 2 , , XI.1970,
Viana col. (MACN 10733). Chubut: Biedma (Puerto Madryn), ,
09.III.1969, Gazztonyi col. (MACN 10741); Florendino Ameghino
(Bahia Aredondu), 2 , I.1976, E. Maury col. (MACN 10739). Figures 37-44. Cybaeodamus brescoviti sp. nov.: 37-39, male palpus: 37, ventral view; 38, prolateral view; 39, retrolateral view; 40, 41,
epigynum: 40, ventral view; 41, dorsal view; Cybaeodamus enigmaticus (Mello Leitão, 1939): 42-44, male palpus: 42, ventral view; 43,
prolateral view; 44, retrolateral view. Iheringia, Sér. Cybaeodamus tocantins sp. nov.
(Figs. 60-67, 68) Types. Holotype
, BRAZIL, Tocantins: Palmas, X.2001,
D. Pavan col. (MZSP 21151). Paratypes: 19 , , Palmas, II.2001-
X.2001, D. Pavan col. (MZSP 21152-21156); Mato Grosso:
,
Chapada dos Guimarães, 20-28.IX.2000, C. Strüssmann col. (MCTP
11567). Etymology. The specific name is a noun in
apposition to the genus name and refers to the type
locality. Diagnosis. Males of C. tocantins sp. nov. differ
from the ones of the other species of the genus by the
presence of a boss on leg IV (Fig. 62), tibial apophysis
elongate and pointed distally on retrolateral view (Fig. 65) and concave on prolateral side (Fig. 63). Females differ
from the ones of the other species of the genus by the
shape of the epigynum, with a median longitudinal atrium
(Fig. 66). Spermatecae reniform (Fig. 67). Female (paratype, MZSP 21152). Total length 5.30. Carapace 2.46 long, 1.48 wide, 0.88 height. Sternum 1.14
long, 0.90 wide. Carapace light chestnut with a large
central longitudinal yellowish spot. Fovea longitudinal,
dark chestnut. Chelicerae dark chestnut, densely coated
with setae. Endites yellow, bristly. Labium chestnut
bristly. Sternum light chestnut, oval, bristly. Ocular area
blackish, bristly with a median longitudinal row of Description. Male (holotype). Total lenght 4.80. Carapace as in the other species, 2.56 long, 1.71 wide,
0.60 height. Sternum 1.30 long, 1.00 wide. Carapace (Fig. 60) yellow, lateral and inferior margins dark chestnut. Fovea longitudinal, narrow, chestnut very dark. Chelicerae chestnut, bristly. Endites whitish yellow,
bristly. Labium chestnut, bristly. Sternum yellow, bordered
with light chestnut, apex dark chestnut, with long strong Figures 45-47. Cybaeodamus enigmaticus (Mello Leitão, 1939): 45, palpus, ventral view; 46, patch of the ventral face of the male
abdomen; 47, idem, detail. Figures 45-47. Cybaeodamus enigmaticus (Mello Leitão, 1939): 45, palpus, ventral view; 46, patch of the ventral face of the male
abdomen; 47, idem, detail. Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 269 On the Neotropical genus Cybaeodamus (Araneae, Zodariidae... remaining articles fulvous. Legs III and IV very spinous. Abdomen 2.63 long, 1.81 wide, 1.48 height. Dorsum
yellowish, shadowed with black. Laterals and venter yellow,
epigynum dark chestnut. Spinnerets light fulvous. Epigynum with median longitudinal atrium (Fig. 66). Variation. Males (10). Total length: 4.00-5.00;
carapace: 2.00-2.69. macrosetae. Leg formula 4,1,3,2. Measurements I/II/III/
IV: femur 1.34/1.22/1.24/1.50; patella 0.64/0.64/0.60/0.68;
tibia 0.96/0.76/0.60/1.00; metatarsus 0.84/0.80/0.94/1.50;
tarsus 0.64/0.60/0.68/0.80. Total 4.42/4.02/4.06/5.48. Figures 48-53. Cybaeodamus lycosoides (Nicolet, 1849): 48, 49, epigynum: 48, ventral view; 49, dorsal view; Cybaeodamus ornatus
Mello-Leitão, 1938: 50, 51, male palpus: 50, ventral view; 51, retrolateral view; 52, 53, epigynum: 52, ventral view; 53, dorsal view. Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 Variation. Males (10). Total length: 4.00-5.00;
carapace: 2.00-2.69. Type. Holotype (examined, immature), ARGENTINA,
Catamarca, Belén, II-III.1939, M. Birabén col. (MLP 14658). Type. Holotype (examined, immature), ARGENTINA,
Catamarca, Belén, II-III.1939, M. Birabén col. (MLP 14658). Type. Holotype (examined, immature), ARGENTINA,
La Rioja, Sañogasta, II-III.1939, M. Birabén col. (MLP 14660). Cybaeodamus tocantins sp. nov.
(Figs. 60-67, 68) Legs
I, II and III with femura yellow on the ectal and ventral
faces shadowed with gray. Remaining articles fulvous. Legs IV with femura ectal end ventral faces whitish yellow, remaining articles fulvous. Legs III and IV very spinous. Abdomen 2.63 long, 1.81 wide, 1.48 height. Dorsum
yellowish, shadowed with black. Laterals and venter yellow,
epigynum dark chestnut. Spinnerets light fulvous. Epigynum with median longitudinal atrium (Fig. 66). remaining articles fulvous. Legs III and IV very spinous. Abdomen 2.63 long, 1.81 wide, 1.48 height. Dorsum
yellowish, shadowed with black. Laterals and venter yellow,
epigynum dark chestnut. Spinnerets light fulvous. Epigynum with median longitudinal atrium (Fig. 66). Variation. Males (10). Total length: 4.00-5.00;
carapace: 2.00-2.69. 270 LISE et al. Valcheta rastellifera MELLO-LEITÃO, 1940:20, fig. 18-20 (described
in Agelenidae).
C b
d
ll f
J
1991 50 Other material examined. BRAZIL. Tocantins: Palmas,
2 , X.2001, D. Pavan col. (MCTP 19987). Mato Grosso: Chapada
dos Guimarães,
, 20-28.IX.2001, C. Strüssmann col. (MCTP
19988). Other material examined. BRAZIL. Tocantins: Palmas,
2 , X.2001, D. Pavan col. (MCTP 19987). Mato Grosso: Chapada
dos Guimarães,
, 20-28.IX.2001, C. Strüssmann col. (MCTP
19988). Type. Holotype (examined, immature), ARGENTINA,
Río Negro, 12-III.1938, M. Birabén col. (MLP 14305). Cybaeodamus pallidus (Mello-Leitão, 1943),
species inquirenda Discussion. MELLO-LEITÃO (1941) described a male
but the holotype examined is a juvenile female. The palpus
in MELLO-LEITÃO (1941: fig. 17) seems not to be of
Cybaeodamus and the picture 13 on plate IV (Mello-Leitão
op. cit.) seems to be of a female. In the Museo de La
Plata there is no additional specimens of C. scottae. Cybaeodamus rastellifer (Mello-Leitão, 1940),
species inquirenda Distribution. North and Central west region of
Brazil (Tocantins and Mato Grosso). Distribution. North and Central west region of
Brazil (Tocantins and Mato Grosso). Valcheta pallida MELLO-LEITÃO, 1943:106. Valcheta pallida MELLO-LEITÃO, 1943:106. Type. Holotype
(probably lost, not examined),
ARGENTINA, Mendoza, Los Molles. Figures 60-67. Cybaeodamus tocantins sp. nov.: 60, male, dorsal view; 61, male carapace, ventral view; 62, femur of leg III; 63-65, male
palpus; 63, ventral view; 64, prolateral view; 65, retrolateral view; 66, 67, epigynum: 66, ventral view; 67, dorsal view. Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 REFERENCES JOCQUÉ, R. 1991. A generic revision of the spider family Zodariidae
(Araneae). Bulletin of the American Museum of Natural
History 201:1-160. LEHTINEN, P. T. 1967. Classification of the cribellate spiders and
some allied families, with notes on the evolution of the
suborder Araneomorpha. Annales Zoologici Fennici 4:199-468. p
g
MELLO-LEITÃO, C. F. 1938. Algunas arañas nuevas de la Argentina. Revista del Museu de La Plata (N.S., Zool.) 1:89-118. 1939 Araignées américaines du Musee d’histoire naturelle Revista del Museu de La Plata (N.S., Zool.) 1:89 118. ___. 1939. Araignées américaines du Musee d’histoire naturelle
de Bâle. Revue suisse Zoology 46:43-93. ___. 1939. Araignées américaines du Musee d histoire naturelle
de Bâle. Revue suisse Zoology 46:43-93. 1940
A
d
l
i
i
d
B
Ai
d
l ___. 1940. Arañas de la provincia de Buenos Aires y de las
gobernaciones de La Pampa, Neuquén, Río Negro y Chubut. Revista del Museu de La Plata (N.S., Zool.) 2:3-62. (
,
)
___. 1941. Las arañas de Córdoba, La Rioja, Catamarca, Tucumán,
Salta y Jujuy colectadas por los Profesores Birabén. Revista
del Museu de La Plata (N.S., Zool.) 2:99-198. (
,
)
___. 1943. Arañas nuevas de Mendoza, La Rioja y Córdoba
colectadas por el Professor Max Birabén. Revista del Museu
de La Plata (N.S., Zool.) 3:101-121. NICOLET, A. C. 1849. Aracnidos. In: GAY, C. ed. Historia física y
política de Chile. Zoología 3:319-543. PLATNICK, N. I. 2007. The world spider catalog, version 7.5. American Museum of Natural History. Avaliable at: <http://
research.amnh.org/entomology/spiders/catalog/index.html>. Accessed on: 16.05.2007. ROTH, V. D. 1965. Genera erroneously placed in the spider families
Agelenidae and Pisauridae (Araneida: Arachnida). Annals of
the Entomological Society of America 58:289 292 ROTH, V. D. 1965. Genera erroneously placed in the spider families
Agelenidae and Pisauridae (Araneida: Arachnida). Annals of
the Entomological Society of America 58:289-292. ___. 1967. A review of the South American spiders of the family
Agelenidae (Arachnida, Araneae). Bulletin of the American
Museum of Natural History 134:297-346 Figure 68. Distribution of Cybaeodamus species in South America. the Entomological Society of America 58:289-292. ___. 1967. A review of the South American spiders of the family the Entomological Society of America 58:289-292. ___. 1967. A review of the South American spiders of the family
Agelenidae (Arachnida, Araneae). Bulletin of the American
Museum of Natural History 134:297-346. Acknowledgments. Type. Holotype
(probably lost, not examined),
ARGENTINA, Mendoza, Los Molles. Type. Holotype
(probably lost, not examined),
ARGENTINA, Mendoza, Los Molles. Figures 54-59. Cybaeodamus ornatus Mello-Leitão, 1938: 54, palpus, ventral view; 55-57, epigynum: 55, 56, ventral view; 57, dors
view; 58 patch of the ventral face of male abdomen; 59 patch of the ventral face of female abdomen Figures 54-59. Cybaeodamus ornatus Mello-Leitão, 1938: 54, palpus, ventral view; 55-57, epigynum: 55, 56, ventral view; 57, dor
view; 58, patch of the ventral face of male abdomen; 59, patch of the ventral face of female abdomen. Figures 54-59. Cybaeodamus ornatus Mello-Leitão, 1938: 54, palpus, ventral view; 55-57, epigynum: 55, 56, ventral view; 57, dorsal
view; 58, patch of the ventral face of male abdomen; 59, patch of the ventral face of female abdomen. Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 On the Neotropical genus Cybaeodamus (Araneae, Zodariidae... 271 Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 LISE et al. 272 Figure 68. Distribution of Cybaeodamus species in South America. Tecnológico” for the Grant (CNPq 140586/2007-5) gave to E. N. L. Rodrigues; to the curators of the collections mentioned for the
loan of material and to Estevam L. C. da Silva for the help on
preparing the final art of the illustrations. Recebido em novembro de 2007. Aceito em dezembro de 2008. ISSN 0073-4721
Artigo disponível em: www.scielo.br/isz Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009 REFERENCES We are grateful to Mónica Tassara of
the Museo de La Plata for the attention; the Direction of Museu de
Ciências Naturais da Fundação Zoobotânica do Rio Grande do Sul
by permitting the third author to develop part of this work in its
dependences; for anonymous reviewers for their valuable comments
on the manuscript; to the staff of Centro de Miscroscopia e
Microanálises da PUCRS, for the help in obtaining the SEM images;
to “Conselho Nacional de Desenvolvimento Científico e Museum of Natural History 134:297-346. SIMON, E. 1889. Etudes arachnologiques. 21e Mémoire. XXX. Descriptions de quelques arachnides du Chili et remarques
synonymiques sur quelques unes des espèces décrites par Nicolet. Annales de la Société Entomologique de France, ser. 6
(8):217-222. Recebido em novembro de 2007. Aceito em dezembro de 2008. ISSN 0073-4721
Artigo disponível em: www.scielo.br/isz Iheringia, Sér. Zool., Porto Alegre, 99(3):259-272, 30 de setembro de 2009
|
https://openalex.org/W3169704303
|
https://www.researchsquare.com/article/rs-611775/latest.pdf
|
English
| null |
Tissue-resident M2 macrophages directly contact primary sensory neurons in the sensory ganglia after nerve injury
|
Journal of neuroinflammation
| 2,021
|
cc-by
| 9,523
|
Tissue-resident M2 Macrophages Directly Contact Primary
Sensory Neurons in the Sensory Ganglia after Nerve Injury Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Tetsuya Goto Tissue-resident M2 Macrophages Directly Contact Primary
Sensory Neurons in the Sensory Ganglia after Nerve Injury
Haruki Iwai
(
haruki@dent.kagoshima-u.ac.jp
)
Kagoshima University Graduate School of Medicine and Dental Sciences: Kagoshima Daigaku Daigakuin Ishigaku
Sogo Kenkyuka
https://orcid.org/0000-0002-2698-3522
Koji Ataka
Kobe Pharmaceutical University: Kobe Yakka Daigaku
Hajime Suzuki
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Ashis Dhar
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Eriko Kuramoto
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Atsushi Yamanaka
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Tetsuya Goto
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Research
Keywords: Neuroinflammation, Nerve injury, Bone-marrow-derived macrophages, Tissue-resident macrophages, M1, M2,
Cell contact, Sensory ganglion, Primary sensory neuron, Satellite glial cell
Posted Date: June 16th, 2021
DOI: https://doi.org/10.21203/rs.3.rs-611775/v1 Tissue-resident M2 Macrophages Directly Contact Primary
Sensory Neurons in the Sensory Ganglia after Nerve Injury
Haruki Iwai
(
haruki@dent.kagoshima-u.ac.jp
)
Kagoshima University Graduate School of Medicine and Dental Sciences: Kagoshima Daigaku Daigakuin Ishigaku
Sogo Kenkyuka
https://orcid.org/0000-0002-2698-3522
Koji Ataka
Kobe Pharmaceutical University: Kobe Yakka Daigaku
Hajime Suzuki
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Ashis Dhar
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Eriko Kuramoto
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Atsushi Yamanaka
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Tetsuya Goto
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Research
Keywords: Neuroinflammation, Nerve injury, Bone-marrow-derived macrophages, Tissue-resident macrophages, M1, M2,
Cell contact, Sensory ganglion, Primary sensory neuron, Satellite glial cell
Posted Date: June 16th, 2021
DOI: https://doi.org/10.21203/rs.3.rs-611775/v1 Tissue-resident M2 Macrophages Directly Contact Primary
Sensory Neurons in the Sensory Ganglia after Nerve Injury
Haruki Iwai
(
haruki@dent.kagoshima-u.ac.jp
)
Kagoshima University Graduate School of Medicine and Dental Sciences: Kagoshima Daigaku Daigakuin Ishigaku
Sogo Kenkyuka
https://orcid.org/0000-0002-2698-3522
Koji Ataka
Kobe Pharmaceutical University: Kobe Yakka Daigaku
Hajime Suzuki
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Ashis Dhar
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Eriko Kuramoto
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Atsushi Yamanaka
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Tetsuya Goto
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Research
Keywords: Neuroinflammation, Nerve injury, Bone-marrow-derived macrophages, Tissue-resident macrophages, M1, M2,
Cell contact, Sensory ganglion, Primary sensory neuron, Satellite glial cell
Posted Date: June 16th, 2021
DOI: https://doi.org/10.21203/rs.3.rs-611775/v1
License: This work is licensed under a Creative Commons Attribution 4 0 International License
Read Full License Tissue-resident M2 Macrophages Directly Contact Primary
Sensory Neurons in the Sensory Ganglia after Nerve Injury
Haruki Iwai
(
haruki@dent.kagoshima-u.ac.jp
)
Kagoshima University Graduate School of Medicine and Dental Sciences: Kagoshima Daigaku Daigakuin Ishigaku
Sogo Kenkyuka
https://orcid.org/0000-0002-2698-3522
Koji Ataka
Kobe Pharmaceutical University: Kobe Yakka Daigaku
Hajime Suzuki
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Ashis Dhar
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Eriko Kuramoto
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Atsushi Yamanaka
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu
Tetsuya Goto
Kagoshima University Graduate School of Medical and Dental Sciences Department of Dentistry: Kagoshima Daigaku
Shigakubu Research Keywords: Neuroinflammation, Nerve injury, Bone-marrow-derived macrophages, Tissue-resident macrophages, M1, M2,
Cell contact, Sensory ganglion, Primary sensory neuron, Satellite glial cell Posted Date: June 16th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-611775/v1 DOI: https://doi.org/10.21203/rs.3.rs-611775/v1 License:
This work is licensed under a Creative Commons Attribution 4.0 International
License. Read Full License Page 1/21 Page 1/21 Page 1/21 Abstract Background: Macrophages in the peripheral nervous system are key players in the repair of nerve tissue and the
development of neuropathic pain due to peripheral nerve injury. However, there is a lack of information on the origin and
morphological features of macrophages in sensory ganglia after peripheral nerve injury, unlike those in the brain and
spinal cord. We analyzed the origin and morphological features of sensory ganglionic macrophages after nerve ligation
or transection using wild-type mice and mice with bone-marrow cell transplants. Methods: After protecting the head of C57BL/6J mice with lead caps, they were irradiated and transplanted with bone-
marrow-derived cells from GFP transgenic mice. The infraorbital nerve of a branch of the trigeminal nerve of wild-type
mice was ligated or the infraorbital nerve of GFP-positive bone-marrow-cell-transplanted mice was transected. After
immunostaining the trigeminal ganglia, the structures of the ganglionic macrophages, neurons, and satellite glial cells
were analyzed using two-dimensional or three-dimensional images. Results: The number of damaged neurons in the trigeminal ganglia increased from day 1 after infraorbital nerve
ligation. Ganglionic macrophages proliferated from days 3 to 5. Furthermore, the numbers of macrophages increased
from days 3 to 15. Bone-marrow-derived macrophages increased on day 7 after the infraorbital nerve was transected in
the trigeminal ganglia of GFP-positive bone-marrow-cell-transplanted mice but most of the ganglionic macrophages
were composed of tissue-resident cells. On day 7 after infraorbital nerve ligation, ganglionic macrophages increased in
volume, extended their processes between the neurons and satellite glial cells, and contacted these neurons. Most of
the ganglionic macrophages showed an M2 phenotype when contact was observed, and little neuronal cell death
occurred. Conclusions: Most of the macrophages that appear after a nerve injury are tissue-resident, and these make direct
contact with damaged neurons that act in a tissue-protective manner in the M2 phenotype. These results imply that
tissue-resident macrophages signal to neurons directly through physical contact. Background Macrophages are distributed in the peripheral nervous system and are key players in the repair of nerve tissue and the
development of neuropathic pain caused by peripheral nerve injury. They are activated after nerve injury, which leads to
nerve fiber repair and hyperalgesia [1,2]. By contrast, suppression of macrophages, such as pharmacological
manipulation, suppresses hyperalgesia and delays nerve fiber repair after nerve injury [3]. Macrophages occupy nerve
fibers and sensory ganglia in the peripheral nervous system [1]. Those located at the site of the nerve fiber injury as well
as those in sensory ganglia away from the site are activated on nerve fiber injury [1]. For instance, ganglionic
macrophages positive for ionized calcium-binding adaptor molecule 1 (Iba1) proliferate [4] and increase in number
after nerve injury [5,6]. Macrophages are broadly divided into tissue-resident macrophages and bone-marrow-derived (BMD) macrophages [7]. Studies on green-fluorescent-protein-positive bone-marrow-cell-transplanted (GFP-BMT) mice have shown that after
nerve injury the spinal dorsal horn in the central nervous system is infiltrated not only by microglia, a tissue-resident
macrophage, but also by BMD macrophages [8,9]. Moreover, a study of physiological turnover reported that
approximately 80% of BMD macrophages in sensory ganglia are replaced within 3 months [10]. However, radiation
results in infiltration of BMD macrophages into the spinal cord dorsal horn [11], which may also occur in sensory
ganglia. A method to protect the mouse brain from radiation damage has been devised, namely, placing a lead cap on
the mouse’s head [12,13]. We thought it would be useful to observe the trigeminal ganglion located in the lower part of Page 2/21 Page 2/21 the brain using GFP-BMT mice with the head protected by a lead cap to distinguish tissue-resident macrophages in the
ganglion from BMD macrophages after nerve injury. The morphology of macrophages differs depending on tissue and can change depending on activation state. Microglia
in the central nervous system have a ramified process in the normal state and are in a rod or amoeboid shape in the
pathological state [14]. Macrophages in the sensory ganglia of hemiplegic migraine model mice are amoeboid in
shape, like microglia [15]. However, there is a lack of information on the individual morphology of ganglionic
macrophages after nerve injury. Furthermore, ganglionic macrophages become ring-like and surround damaged
neurons in nerve-injury models [16,17]. In addition, ganglionic macrophages contact neurons after nerve injury [5]. Animals This study was approved by the Animal Experiment Committee of Kagoshima University and was conducted according
to the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals. All efforts were made to
minimize the number of animals included. We used 77 male C57BL/6J mice weighing 20–30 g (Japan SLC, Shizuoka,
Japan) and C57BL/6-Tg (UBC-GFP) 30Scha/J (green-fluorescent-protein-transgenic; GFP-Tg) mice (Jackson Laboratory,
Bar Harbor, ME, USA). Surgery was performed under anesthesia induced by an intraperitoneal injection of 0.3 mg/kg
medetomidine (Meiji Seika Pharma, Tokyo, Japan), 4.0 mg/kg midazolam (Novartis, Basel, Switzerland), and 5.0 mg/kg
butorphanol (Meiji Seika Pharma). Background As
neurons in healthy sensory ganglia are covered with glutamine-synthetase-positive satellite glial cells [18], the
morphological relationships among macrophages, neurons, and satellite glial cells need to be clarified in three
dimensions if macrophages contact neurons after nerve injury. Macrophages can be classified into either the M1 pro-inflammatory phenotype, which secretes cytokines, or the M2
anti-inflammatory phenotype, which protects tissues [19]. After nerve injury, ganglionic macrophages increase levels of
the M1 marker cluster of differentiation 86 (CD86) and the M2 marker cluster of differentiation 206 (CD206) [20], or
CD206 alone [6, 21,22]. However, it is unclear whether macrophages should be classified as the M1 or M2 phenotype
and phagocytose neurons when they come in contact with neurons. In this study, we clarified the origin and three-dimensional morphological changes in macrophages after chronologically
organizing their activation patterns in sensory ganglia after nerve injury. We examined when macrophages reach their
peak of division, whether tissue-resident or BMD macrophages dominate, what individual forms the macrophages take
and how they contact damaged neurons, whether macrophages are classified as the M1 or M2 phenotype after contact,
and whether neural cell death occurs. We found that M2 macrophages, which are found mainly in sensory ganglia,
proliferate, increase in volume, and enter areas between primary sensory neurons and satellite glial cells, which make
direct contact with neurons after nerve injury. Animal surgery Animal surgery was performed according to a previous study [23]. In 55 mice (8- to 12-week-old wild-type mice, n = 48;
7-month-old GFP-BMT mice, n = 7), the left side of the infraorbital nerve was the experimental side and the right side
was the sham-operated side. Nineteen mice were used as naïve controls. As a previous study using GFP-BMT mice
reported that 80% of the physiological turnover of macrophages in sensory ganglia occurred within 3 months [10], we
used 7-month-old mice, which was 5 months after the transplantation. The buccal mucosa of the upper jaw on the left
experimental side was small incised and the infraorbital nerve came out from the foramen was exposed. The nerve was
tightly ligated with 7–0 silk suture in wild-type mice. As a previous study using GFP-BMT mice prepared a nerve crush
lesion model [10], the infraorbital nerve was transected to completely damage the entire nerve rather than part of it in
the GFP-BMT mice. After nerve injury, the incision was closed with tissue adhesive. The infraorbital nerve received the
same procedure on the right sham-operated side as on the left side without ligature or transection. After a survival
period of 1 day to 4 weeks, the mice were deeply anesthetized, transcardially perfused with phosphate-buffered saline
(PBS; pH 7.3), and fixed with 4% formaldehyde in phosphate buffer (pH 7.3). Forty-six mice received an intraperitoneal
injection of 100 mg/kg bromodeoxyuridine (BrdU; B5002, Merck, Darmstadt, Germany) in PBS to detect cell proliferation
at 24 h before perfusion. of GFP-positive cells in monocytes was examined in each mouse by flow-assisted cell sorting (Accuri C6; BD
Biosciences, Franklin Lakes, NJ, USA). GFP-BMT mice with a chimeric ratio > 70% were used. of GFP-positive cells in monocytes was examined in each mouse by flow-assisted cell sorting (Accuri C6; BD
Biosciences, Franklin Lakes, NJ, USA). GFP-BMT mice with a chimeric ratio > 70% were used. Immunohistochemistry The trigeminal ganglia were removed, postfixed in the same fixative solution, kept overnight at 4°C, and immersed in
30% sucrose in PBS for 2–3 days at 4°C. Serial horizontal sections of two thicknesses of the trigeminal ganglia were
cut on a cryostat (CryoStar NX70; Thermo Fisher Scientific, Waltham, MA, USA); 30-μm sections were used for normal
histological observation, and 50-μm sections were used for three-dimensional reconstructions. Immunostaining was
divided into two groups, one for bright-field observations and the other for fluorescence observations, and each was
reacted at room temperature. The sections for BrdU immunostaining were submerged in 2 N HCl for 60 min before the
immunoreaction. The sections were incubated with 1% bovine serum albumin and 0.2% Triton X-100 in PBS for 1 h before the
fluorescence observations; primary antibodies were incubated overnight in the same incubation buffer, and secondary
antibodies were incubated in the same incubation buffer for 3 h. Subsequently, the sections received nuclear staining of
1:100 4,’6-diamidino-2-phenylindole (DAPI; 340–07971, Dojindo, Kumamoto, Japan) or Nissl staining with 1:1000
NeuroTrace 500/525 (N-21480, Thermo Fisher Scientific) in PBS. The antibodies used are shown in Table 1. The sections for bright-field observations were incubated with 3% normal rabbit serum and 0.2% Triton X-100 in PBS for
1 h; primary antibodies were incubated in the same incubation buffer overnight, and secondary antibodies were
incubated in the same incubation solution for 3 h. Then the sections were incubated in 1:100 avidin-biotin-peroxidase
complex (PK-4000, Vector Laboratories, Burlington, CA, USA) in PBS for 1 h. Subsequently, the sections were reacted
with 0.02% diaminobenzidine tetrahydrochloride (D5637, Merck, Darmstadt, Germany) and 0.005% H2O2 in 0.05 M Tris-
HCl buffer for 20 min. Finally, the sections were mounted on glass slides (Platinum Pro; Matsunami Glass, Osaka, Japan), air-dried, and
coverslipped. Photomicrographs of the ganglion sections were taken using a digital slide scanner (BZ-X700; Keyence,
Osaka, Japan) or a confocal laser scanning microscope (LSM 700; Carl Zeiss Microscopy, Jena, Germany). Data analysis Bone marrow transplantation Bone-marrow transplantation was performed according to previous studies [12,13]. Bone-marrow cells were isolated
from the femurs and tibias of GFP-Tg mice (n = 3). Recipient 7- to 9-week-old C57BL/6J mice (n = 7) with the head
covered with a lead cap received whole-body irradiation of 10 Gy, and bone-marrow cells (4–6 × 106) were injected into
their tail veins. GFP-BMT mice were maintained in cages, covered with filter caps, and given sterile water including
0.001 N HCl (pH 2.0) and sterile chow for 2 weeks to prevent infection. Eight to ten weeks after transplantation, the ratio Page 3/21 Page 3/21 Proliferation of ganglionic macrophages after nerve injury To evaluate neuronal damage and cellular activity of macrophages in sensory ganglia after nerve injury over time, the
maxillary nerve region of the trigeminal ganglion was immunohistochemically stained after ligating the infraorbital
nerve. Activating transcription factor 3 (ATF3) is a marker of damaged neurons [24] and is also involved in axonal
regeneration, as axonal outgrowth is reduced in ATF3-deficient mice [25]. Therefore, ATF3 was used to identify the
location to observe in the maxillary nerve region of the trigeminal ganglia and to confirm the expression of ATF3 in the
time series. After the infraorbital nerve was ligated, ATF3-positive cells were detected over a wide range of the maxillary
nerve region on the ipsilateral side (Fig. 1a). By contrast, the mandibular nerve region in the ipsilateral trigeminal
ganglia had almost no ATF3-positive cells (data not shown). Therefore, the entire maxillary nerve region was used as
our observation point. The numbers of ATF3-positive cells increased significantly on the ipsilateral side of the maxillary
nerve region compared to the contralateral side from day 1 to day 15 after infraorbital nerve ligation (Fig. 1a, b). The macrophage/microglia marker Iba1 [26,27] was used to examine changes in the numbers of macrophages in a
time series. After the infraorbital nerve was ligated, the numbers of Iba1-positive cells increased significantly on the
ipsilateral side of the maxillary nerve region compared to the contralateral side from day 3 to day 15 (Fig. 1c, d). The cell-proliferation marker BrdU was administered as a single dose to nerve-ligated mice 24 h before perfusion. After
infraorbital nerve ligation, numbers of BrdU- and Iba1-positive cells increased significantly on the ipsilateral side of the
maxillary nerve region compared to the contralateral side from day 3 to day 5 (Fig. 1e–g). These results imply that
primary sensory neurons were damaged on day 1, and then ganglionic macrophages underwent cell proliferation from
days 3 to 5 and cell numbers increased from days 3 to 15 after nerve injury. Data analysis Page 4/21 Page 4/21 Measurements of ganglionic sections were analyzed using Fiji/ImageJ software (NIH, Bethesda, MD, USA). Three-
dimensional reconstructed images were drawn using Neurolucida software (MBF Bioscience, Williston, VT, USA) from
photomicrographs taken under a confocal laser scanning microscope. The statistical analysis employed Graphpad
Prism 7 software (GraphPad Software, La Jolla, CA, USA). Data are expressed as mean ± standard deviation, and
differences were detected using one-way or two-way factorial analysis of variance (ANOVA) with the Tukey–Kramer
post hoc test or the unpaired Student’s t-test with Welch’s correction. A P-value < 0.05 was considered significantly
different. Morphological changes in ganglionic macrophages after nerve injury To investigate changes in the form of macrophages in sensory ganglia after nerve injury, we attempted to three-
dimensionally reconstruct macrophages in the maxillary nerve region of the trigeminal ganglia after ligating the
infraorbital nerve. Before reconstruction, we examined the increase in the cell area of ganglionic macrophages over time after ligating the
infraorbital nerve. That of Iba1-positive cells increased significantly on the ipsilateral side of the maxillary nerve region
compared to the contralateral side from day 5 to day 15 after infraorbital nerve ligation (Fig. 3a, b). Subsequently, the volume of macrophages around damaged neurons was measured on day 7, when the cell area of
macrophages was at its maximum after ligation. After ligation, the cell volume of Iba1-positive cells surrounding ATF3-
positive cells on the ipsilateral side increased significantly compared to Iba1-positive cells surrounding ATF3-negative
cells on the contralateral side (Fig. 3c–e). In addition, Iba1-positive cells appeared to extend multiple processes that
surrounded ATF3-positive cells (Fig. 3d). Before three-dimensional reconstruction of macrophages, neurons, and satellite glial cells that contacted after
infraorbital nerve ligation, we defined “a contact-like structure” as a structure in which macrophages appeared to be in
contact with neurons, and studied this structure over time. We used ATF3, Iba1, and product gene protein 9.5 (PGP9.5; a
neuron marker) as indicators and prepared a simplified classification as follows: ATF3-positive neurons form a contact-
like structure with Iba1-positive cells (ATF3+CLS+ neurons), ATF3-negative neurons form a contact-like structure with
Iba1-positive cells (ATF3-CLS+ neurons), ATF3-positive neurons do not form a contact-like structure with Iba1-positive
cells (ATF3+CLS- neuron), and ATF3-negative neurons do not form a contact-like structure with Iba1-positive cells (ATF-
CLS- neuron). Most of the neurons showed contact-like structures with Iba1-positive cells from day 5 to day 7 after
infraorbital nerve ligation (Fig. 4a, b). In naïve mice, neurons in sensory ganglia are covered with satellite glial cells [18]. To ascertain whether ganglionic
macrophages contact the neurons and to determine the area of contact after nerve injury, we created three-dimensional
reconstructed images from the immunohistochemistry results, and measured the contact areas between neurons and
macrophages or satellite glial cells. We defined Iba1-positive cells, which are located between neurons and GS-positive
cells, as macrophages contacting neurons (Fig. 4c). Reconstruction was performed on day 7, when the contact-like
structures between macrophages and neurons were observed (Fig. 4a, b) and the area of macrophages was at its
maximum (Fig. 3a, b). Migration of ganglionic macrophages after nerve injury To elucidate whether proliferated macrophages in sensory ganglia after nerve injury are tissue-resident or BMD, we
transected the infraorbital nerve of head-protected GFP-BMT mice and looked for GFP-positive BMD cells in the
maxillary nerve region of the trigeminal ganglia. After radiation exposure, macrophages in the dorsal horn of the spinal cord migrate from blood vessels due to
disruption of the blood–brain barrier [11]. Therefore, in this study, the head, including the trigeminal ganglion, was
protected with a lead cap to prevent direct radiation exposure. The body of head-protected GFP-BMT mice turned white
and the head turned black (Fig. 2a). Macrophages in sensory ganglia are thought to originate from bone marrow [10]. To confirm this, we created a nerve-
injury model in 7-month-old mice, 5 months after transplanting BMD cells. As the macrophages barely proliferated and Page 5/21 cell numbers peaked and remained constant on day 7 after nerve injury (Fig. 1c–g), we chose day 7 as our observation
point. In addition, we employed infraorbital nerve transection to damage the entire infraorbital nerve rather than part of
it. After transecting the infraorbital nerve, the numbers of GFP-positive cells increased significantly on the ipsilateral
side of the maxillary nerve region compared to the contralateral side by day 7 (Fig. 2b, d). Similarly, the numbers of
GFP- and Iba1-positive cells increased significantly on the ipsilateral side by day 7 after nerve transection (Fig. 2b, c, e). Furthermore, GFP-negative and Iba1-positive cells made up the majority of cells (Fig. 2b, f). These results imply that
although the number of BMD macrophages increased after nerve injury, most of the ganglionic macrophages were
tissue-resident. Discussion Macrophages in sensory ganglia are essential for controlling inflammation, nerve tissue repair and neuropathic pain
after peripheral nerve injury. We showed that ganglionic macrophages, which proliferate rapidly after nerve injury, are
tissue-resident, enter between satellite glial cells and neurons, are in direct contact with neurons, and exhibit the M2
phenotype with almost no neuronal cell death, indicating that this contact may be related to tissue repair. Until now,
whether ganglionic macrophages that increase after peripheral nerve injury are tissue-resident or BMD has been studied
only indirectly, such as with macrophage-depleting agents, and no direct evidence has been reported. In addition, while
morphological changes in microglia after peripheral nerve injury are an indicator of activation, ganglionic macrophages
lack morphological information, including a three-dimensional structure. Our study was designed to compensate for
this. Morphological changes in ganglionic macrophages after nerve injury Neurons were covered with glutamine-synthetase-positive cells, and Iba1-positive cells did not
contact neurons on the contralateral side on day 7 after infraorbital nerve ligation (Fig. 4d,e), whereas Iba1-positive
cells directly contacted the neurons on the ipsilateral side (Fig. 4c–e). The percentage of surface area where Iba1-
positive cells were in contact with neurons was about 13.71% (Fig. 4f, g). These results indicate that ganglionic
macrophages increase in volume, and directly contact damaged neurons via multiple processes after nerve injury. Page 6/21 Page 6/21 Classification of ganglionic macrophages after nerve injury Macrophages are broadly classified into the M1 or M2 phenotypes [19]. To determine whether the macrophages that
contacted neurons were of the M1 or M2 phenotype in sensory ganglia on day 7 after nerve injury, we investigated
CD86, an M1 phenotype marker, and CD206, an M2 phenotype marker, in the maxillary nerve region of the trigeminal
ganglia after ligating the infraorbital nerve. The numbers of CD206- and Iba1-positive cells increased significantly on
the ipsilateral side compared to the contralateral side (Fig. 5a–c). By contrast, the numbers of CD86- and Iba1-positive
cells did not increase significantly (Fig. 5d, f), but a zone rich in CD86- and Iba1-positive cells was observed in some
areas (Fig. 5e). Then we examined whether contact between ganglionic macrophages and neurons was associated with neuronal cell
death after nerve injury. Contact between macrophages and neurons was seen on day 7 after nerve injury (Fig. 4c–g),
and previous study by Western blotting has shown that caspase-3, a marker of cell death, is elevated from week 1 to
week 4 after nerve injury [28]. Therefore, to determine the number of dying neurons after nerve injury, we performed a
histological study using cleaved caspase-3. Caspase-3-positive cells were detected on the ipsilateral side at 1 week
after infraorbital nerve ligation (Fig. 5g) and then decreased in number at from 2 to 4 weeks after infraorbital nerve
ligation (Fig. 5h). Small sensory ganglia neurons are more likely to die after nerve injury [29,30]. Therefore, we measured neuron size to
determine whether caspase-3-positive cells were more common in small or large neuron types. The soma size of
caspase-3- and PGP9.5-positive neurons was < 25 µm at 1 week after infraorbital nerve ligation (Fig. 5i, j). These results
show that ganglionic macrophages exhibited the M2 phenotype on day 7 after nerve injury when macrophages were in
contact with neurons, and that cell death occurred in a small number of damaged small neurons but not in most
damaged neurons. MD macrophages increase in sensory ganglia after nerve injury, but most macrophag We found that BMD macrophages increased in sensory ganglia after nerve injury, but the majority of ganglionic
macrophages were tissue-resident. In a previous study of physiological turnover using bone-marrow-transplanted mice
in sensory ganglia, 80% of cells were replaced by BMD macrophages within 3 months [10]. However, under the influence
of radiation BMD macrophages infiltrate the central nervous system [11], so we protected the head of mice with a lead
cap during irradiation and observed the trigeminal ganglia located in the lower part of the brain. In this way, we
determined that most of the ganglionic macrophages were tissue-resident in mice at 5 months after cell
transplantation, implying that the turnover of BMD cells in sensory ganglia may be due to radiation damage. This result
supports reports that the use of the macrophage-depleting agent clodronate does not change the numbers of
macrophages in sensory ganglia but reduces the numbers of monocytes in the blood [3,4]. Considering that tissue-
resident macrophages in the central nervous system are microglia [7], macrophages in the sensory ganglia of the
peripheral nervous system may be tissue-resident. Nevertheless, an increase in BMD macrophages has also been
observed after nerve injury. Cobos et al. [33] reported a decrease in mRNA for the monocyte/macrophage markers
CD68, CD11b, and CD 163 in sensory ganglia after nerve injury by clodronate treatment. The decrease in these markers
implies that some macrophages in sensory ganglia are BMD macrophages. Macrophages proliferate in sensory ganglia after nerve injury We identified a time series of phenomena associated with inflammation in sensory ganglia, such as an increase in the
numbers of damaged neurons from day 1 after peripheral nerve injury followed by division of macrophages from days
3 to 5 and an increase in the numbers of macrophages on days 3 to 15. Previous histological studies on ATF3 in
sensory ganglia after nerve injury have reported a rapid increase in ATF-positive cells from day 1 [24,31]. This trend was
also observed in an investigation of ATF3-positive cell numbers in the facial nucleus after nerve injury [25]. Krishnan et
al. [4] examined the proliferation of ganglionic macrophages after nerve injury with BrdU and Ki67 and reported a
significant increase on day 3. In their preliminary study using Ki67, although there was no significant difference, the
increase began on day 2, peaked on day 3, and showed a decreasing trend after day 7. Taken together, our results imply Page 7/21 Page 7/21 that ganglion macrophages proliferate from days 3 to 5 after nerve injury, and probably do not persist until day 7. Lu
and Richardson [32] reported that, after nerve injury, ganglionic macrophages increase within 4 days and persist for
more than 1 month. Kwon et al. [5] reported an increase in the numbers of ganglionic macrophages from 3 to 28 days
after nerve injury. Our results support these findings. In brief, ganglionic macrophages proliferate and increase rapidly
after neurons are damaged due to nerve injury, and this state seems to be maintained. Conclusions We found that, in sensory ganglia, macrophages proliferate for several days after neurons are damaged, most
macrophages in ganglia are tissue-resident, macrophages become larger and directly contact damaged neurons, most
macrophages are of the M2 phenotype, and neuronal cell death rarely occurs. The significance of the contact of
macrophages with neurons is unknown and awaits further study. Macrophages presented the M2 phenotype in sensory ganglia after nerve injury Ganglionic macrophages showed M2 phenotype activity in sensory ganglia and a small number of small neurons died
after the nerve injury. Upregulation of the M2 markers CD206 and arginase-1 after nerve injury has been reported
previously [6,20–22]. The expression of CD206 in our study was similar to these previous results. However, although
M1 marker CD86-positive macrophages were not conspicuous throughout sensory ganglia, CD86-positive
macrophages were partially accumulated. Niemi et al. [20] reported an increase in CD86 mRNA along with CD206, and
their data represent the small number of CD86-positive macrophages observed in this study. The significance of the
rare accumulation of CD86-positive macrophages is unclear and awaits further investigation. If we assume that most ganglionic macrophages after nerve injury are neuroprotective, neuronal cell death may be
unlikely to occur. In previous studies, a significant decrease in the numbers of cells was observed at 7 days after nerve
injury [36], whereas neuronal cell death was not observed [37,38] or was observed only in a few neurons by TUNEL
assay [39,40]. In this study, we used caspase-3 and detected a small number of caspase-3-positive neurons; these cells
decreased from weeks 1 to 4. This same decrease was also observed in western blotting of sensory ganglia [28]. The
numbers of caspase-3-positive neurons were considerably lower than the numbers of ATF3-positive neurons after nerve
injury. It is conceivable that most damaged neurons do not die after distal peripheral nerve injury, such as that in this
study. On the other hand, some caspase-3-positive cells were observed until at least 3 weeks, implying that cells died
gradually. In addition, caspase-3 tended to be detected in small neurons. Previous studies have found that small cells
are more likely to die after nerve injury [29,30] and unmyelinated fibers are more likely to disappear than are myelinated
fibers [29,41]. Furthermore, Vega-Avelaira et al. [16] reported that many ganglionic macrophages around C-neurons (<
25 µm) do not have ring-like structures whereas A-neurons (> 25 µm) do. These findings imply that after nerve injury
vigorous contact with M2 phenotype macrophages helps to protect neurons from neuronal cell death rather than
phagocytosing neurons. Morphological changes in macrophages from sensory ganglia after nerve injury We found that the area of macrophages increased from day 5 to day 15 after nerve injury. In addition, we also extracted
and three-dimensionally reconstructed macrophages after nerve injury on day 7 and found that the cytoplasm of
macrophages around damaged nerve cells increased. Franceschini et al. [15] reported an increase in the volume of
macrophages in the trigeminal ganglia of hemiplegic migraine model mice, and our results are similar, although the
model was different. Moreover, Franceschini et al. [15] showed that ganglionic macrophages are amoeboid-like
microglia in the central nervous system, but in our study, they extended processes out to surround the neurons. We
analyzed the ganglionic macrophages adjacent to ATF3-positive neurons, and these morphological differences may
have been due to differences in the models used and the sites measured. Most of the neurons showed contact-like structures with macrophages from days 5 to 7 after nerve injury. These
structures were observed in damaged and undamaged neurons. Furthermore, we extracted and three-dimensionally
reconstructed the cells on day 7 after nerve injury and found that the average contact rate of macrophages to the
neuronal surface was 13.71%. Kwon et al. [5] reported that approximately 75% of ganglionic macrophages are in
contact with neurons after nerve injury. In addition, we showed that the contact-like structures of macrophages
extended to undamaged neurons, implying that ganglionic macrophages affect neurons that are not directly damaged. Neurons in sensory ganglia that have not been injured are covered with satellite glial cells [18], and in our study neurons
were surrounded by satellite glial cells in the three-dimensional reconstruction of the sham side. By contrast, neurons
on the injured side were not completely covered by satellite glial cells; macrophages partially entered between neurons
and satellite glial cells, and neurons and macrophages came in contact with each other. In the central nervous system,
contact between microglia and the cell bodies of neurons is protective, inhibiting the over-activity that occurs during
neuronal injury [34,35]. It is unclear whether these same functions are present in ganglionic macrophage-neuron
contacts, and further investigation is needed. Page 8/21 List Of Abbreviations ATF3: Activating transcription factor 3; BrdU: Bromodeoxyuridine; BMD: Bone marrow-derived; CD86: Cluster of
differentiation 86; CD206: Cluster of differentiation 206; CLS: contact-like structure; Contra: contralateral; DAPI: 4',6-
diamidino-2-phenylindole; GFP: Green fluorescent protein; GFP-BMT: green fluorescent protein-positive bone marrow
cells-transplanted; GFP-Tg: green fluorescent protein-transgenic; Iba1: Ionized calcium-binding adaptor molecule 1; Ipsi:
ipsilateral; PBS: Phosphate buffered saline; PGP9.5: Product gene protein 9.5 Availability of data and materials The datasets used and/or analysed during the current study are available from the corresponding author on reasonable
request. Consent for publication Not applicable. Authors' contributions HI, EK, AY, and TG designed the research; HI, KA, HS, and AD performed the research; HI analyzed the data; HI wrote the
paper. The authors read and approved the final manuscript. Funding This work was supported by Grant-in-Aid for Scientific Research (C) (20K09881 to H.I., 16K11454 and 19K10058 to
E.K.) and the Supporting program for Young Researcher in Kagoshima University. Ethics approval and consent to participate The present study was approved by the Animal Experiment Committee of Kagoshima University and performed in
accordance with the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals. Page 9/21 Page 9/21 Authors' information 1 Department of Oral Anatomy and Cell Biology, Graduate School of Medical and Dental Sciences, Kagoshima
University, 8-35-1 Sakuragaoka, Kagoshima, Kagoshima 890-8544, Japan. 2 Department of Psychosomatic Internal
Medicine, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima,
Kagoshima 890-8544, Japan. 3 Laboratory of Medical Biochemistry, Kobe Pharmaceutical University, 4-19-1
Motoyamakita-machi, Higashinada-ku, Kobe, 658-8558, Japan. 4 Department of Oral and Maxillofacial Surgery,
Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima, Kagoshima
890-8544, Japan. Acknowledgements We wish to thank Joint Research Laboratory, Kagoshima University Graduate School of Medical and Dental Sciences,
for the use of their facilities, and the Facility of Laboratory Animal Science Research Support Center Institute for
Research Promotion Kagoshima University. Competing interests The authors declare that they have no competing interests. References 1. Zigmond RE, Echevarria FD. Macrophage Biology in the Peripheral Nervous System after Injury. Prog Neurobiol. 2018;173:102–21. 1. Zigmond RE, Echevarria FD. Macrophage Biology in the Peripheral Nervous System after Injury. Prog Neurobiol. 2018;173:102–21. 1. Zigmond RE, Echevarria FD. Macrophage Biology in the Peripheral Nervous System after Injury. Prog Neurobiol. 2018;173:102–21. 2. Ji R-RR, Chamessian A, Zhang Y-QQ. Pain regulation by non-neuronal cells and inflammation. Science (New York,
NY). 2016;354:572–7. 2. Ji R-RR, Chamessian A, Zhang Y-QQ. Pain regulation by non-neuronal cells and inflammation. Science (New York,
NY). 2016;354:572–7. 2. Ji R-RR, Chamessian A, Zhang Y-QQ. Pain regulation by non-neuronal cells and inflammation. Science (New York,
NY). 2016;354:572–7. 3. Peng J, Gu N, Zhou L, Eyo UB, Murugan M, Gan W-B, et al. Microglia and monocytes synergistically promote the
transition from acute to chronic pain after nerve injury. Nature Communications. 2016;7:ncomms12029. 3. Peng J, Gu N, Zhou L, Eyo UB, Murugan M, Gan W-B, et al. Microglia and monocytes synergistically promote the
transition from acute to chronic pain after nerve injury. Nature Communications. 2016;7:ncomms12029. Page 10/21 4. Krishnan A, Bhavanam S, Zochodne D. An Intimate Role for Adult Dorsal Root Ganglia Resident Cycling Cells in the
Generation of Local Macrophages and Satellite Glial Cells. Journal of neuropathology and experimental neurology. 2018;77:929–41. 4. Krishnan A, Bhavanam S, Zochodne D. An Intimate Role for Adult Dorsal Root Ganglia Resident Cycling Cells in the
Generation of Local Macrophages and Satellite Glial Cells. Journal of neuropathology and experimental neurology. 2018;77:929–41. 5. Kwon M, Kim J, Shin H, Jeong S, Kang Y, Choi J, et al. Contribution of Macrophages to Enhanced Regenerative
Capacity of Dorsal Root Ganglia Sensory Neurons by Conditioning Injury. The Journal of Neuroscience. 2013;33:15095–108. 6. Kwon M, Shin H, Cui Y, Kim H, Thi A, Choi J, et al. CCL2 Mediates Neuron–Macrophage Interactions to Drive
Proregenerative Macrophage Activation Following Preconditioning Injury. The Journal of Neuroscience. 2015;35:15934–47. 7. Zhao Y, Zou W, Du J, Zhao Y. The origins and homeostasis of monocytes and tissue‐resident macrophages in
physiological situation. J Cell Physiol. 2018;233:6425–39. 8. Echeverry S, Shi XQ, Rivest S, Zhang J. Peripheral Nerve Injury Alters Blood–Spinal Cord Barrier Functional and
Molecular Integrity through a Selective Inflammatory Pathway. J Neurosci. 2011;31:10819–28. 9. Zhang J, Shi XQ, Echeverry S, Mogil JS, Koninck YD, Rivest S. Expression of CCR2 in Both Resident and Bone
Marrow-Derived Microglia Plays a Critical Role in Neuropathic Pain. J Neurosci. 2007;27:12396–406. References 10. Müller M, Leonhard C, Krauthausen M, Wacker K, Kiefer R. On the longevity of resident endoneurial macrophages in
the peripheral nervous system: a study of physiological macrophage turnover in bone marrow chimeric mice. Journal of
the peripheral nervous system : JPNS. 2010;15:357–65. 11. Tashima R, Mikuriya S, Tomiyama D, Shiratori-Hayashi M, Yamashita T, Kohro Y, et al. Bone marrow-derived cells in
the population of spinal microglia after peripheral nerve injury. Scientific Reports. 2016;6:23701. 12. Sawada A, Niiyama Y, Ataka K, Nagaishi K, Yamakage M, Fujimiya M. Suppression of bone marrow-derived
microglia in the amygdala improves anxiety-like behavior induced by chronic partial sciatic nerve ligation in mice. Pain. 2014;155:1762–72. 13. Ataka K, Asakawa A, Nagaishi K, Kaimoto K, Sawada A, Hayakawa Y, et al. Bone Marrow-Derived Microglia Infiltrate
into the Paraventricular Nucleus of Chronic Psychological Stress-Loaded Mice. Plos One. 2013;8:e81744. 14. Sierra A, Castro F de, Río‐Hortega J del, Iglesias‐Rozas JR, Garrosa M, Kettenmann H. The “Big‐Bang” for modern
glial biology: Translation and comments on Pío del Río‐Hortega 1919 series of papers on microglia. Glia. 2016;64:1801–40. 15. Franceschini A, Vilotti S, Ferrari MD, Maagdenberg A van den, Nistri A, Fabbretti E. TNFα levels and macrophages
expression reflect an inflammatory potential of trigeminal ganglia in a mouse model of familial hemiplegic migraine. PloS one. 2013;8:e52394. 16. Vega-Avelaira D, Géranton SM, Fitzgerald M. Differential regulation of immune responses and macrophage/neuron
interactions in the dorsal root ganglion in young and adult rats following nerve injury. Molecular pain. 2009;5:70. 16. Vega-Avelaira D, Géranton SM, Fitzgerald M. Differential regulation of immune responses and macrophage/neuron
interactions in the dorsal root ganglion in young and adult rats following nerve injury. Molecular pain. 2009;5:70. 17. Bravo‐Caparrós I, Ruiz‐Cantero MC, Perazzoli G, Cronin SJF, Vela JM, Hamed MF, et al. Sigma‐1 receptors control
neuropathic pain and macrophage infiltration into the dorsal root ganglion after peripheral nerve injury. Faseb J. 2020;34:5951–66. 17. Bravo‐Caparrós I, Ruiz‐Cantero MC, Perazzoli G, Cronin SJF, Vela JM, Hamed MF, et al. Sigma‐1 receptors control
neuropathic pain and macrophage infiltration into the dorsal root ganglion after peripheral nerve injury. Faseb J. 2020;34:5951–66. Page 11/21 Page 11/21 Page 11/21 18. Hanani M, Spray DC. Emerging importance of satellite glia in nervous system function and dysfunction. Nat Rev
Neurosci. 2020;21:485–98. 19. Liu P, Peng J, Han G-H, Ding X, Wei S, Gao G, et al. Role of macrophages in peripheral nerve injury and repair. Neural
Regen Res. 2019;14:1335. References 20. Niemi JP, DeFrancesco-Lisowitz A, Cregg JM, Howarth M, Zigmond RE. Overexpression of the monocyte chemokine
CCL2 in dorsal root ganglion neurons causes a conditioning-like increase in neurite outgrowth and does so via a STAT3
dependent mechanism. Exp Neurol. 2016;275:25–37. 21. Komori T, Morikawa Y, Inada T, Hisaoka T, Senba E. Site-specific subtypes of macrophages recruited after peripheral
nerve injury. Neuroreport. 2011;22:911–7. 22. Lindborg JA, Niemi JP, Howarth MA, Liu KW, Moore CZ, Mahajan D, et al. Molecular and cellular identification of the
immune response in peripheral ganglia following nerve injury. J Neuroinflamm. 2018;15:192. 23. Kim YS, Chu Y, Han L, Li M, Li Z, Lavinka PC, et al. Central terminal sensitization of TRPV1 by descending
serotonergic facilitation modulates chronic pain. Neuron. 2014;81:873–87. 24. Tsujino H, Kondo E, Fukuoka T, Dai Y, Tokunaga A, Miki K, et al. Activating Transcription Factor 3 (ATF3) Induction
by Axotomy in Sensory and Motoneurons: A Novel Neuronal Marker of Nerve Injury. Mol Cell Neurosci. 2000;15:170–82. 25. Gey M, Wanner R, Schilling C, Pedro MT, Sinske D, Knöll B. Atf3 mutant mice show reduced axon regeneration and
impaired regeneration-associated gene induction after peripheral nerve injury. Open biology. 2016;6. 26. Imai Y, Ibata I, Ito D, Ohsawa K, Kohsaka S. A Novel Gene iba1in the Major Histocompatibility Complex Class III
Region Encoding an EF Hand Protein Expressed in a Monocytic Lineage. Biochem Bioph Res Co. 1996;224:855–62. 27. Ito D, Imai Y, Ohsawa K, Nakajima K, Fukuuchi Y, Kohsaka S. Microglia-specific localisation of a novel calcium
binding protein, Iba1. Mol Brain Res. 1998;57:1–9. 28. Wiberg R, Novikova LN, Kingham PJ. Evaluation of apoptotic pathways in dorsal root ganglion neurons following
peripheral nerve injury. Neuroreport. 2018;29:779–85. 29. Lekan HA, Chung K, Yoon YW, Chung JM, Coggeshall RE. Loss of dorsal root ganglion cells concomitant with dorsal
root axon sprouting following segmental nerve lesions. Neuroscience. 1997;81:527–34. 30. Vestergaard S, Tandrup T, Jakobsen J. Effect of permanent axotomy on number and volume of dorsal root ganglion
cell bodies. J Comp Neurol. 1997;388:307–12. 31. Zhang M-D, Barde S, Yang T, Lei B, Eriksson LI, Mathew JP, et al. Orthopedic surgery modulates neuropeptides and
BDNF expression at the spinal and hippocampal levels. Proc National Acad Sci. 2016;113:E6686–95. 32. Lu X, Richardson PM. Responses of macrophages in rat dorsal root ganglia following peripheral nerve injury. J
Neurocytol. 1993;22:334–41. 33. Cobos EJ, Nickerson CA, Gao F, Chandran V, Bravo-Caparrós I, González-Cano R, et al. References Mechanistic Differences in
Neuropathic Pain Modalities Revealed by Correlating Behavior with Global Expression Profiling. Cell Reports. 2018;22:1301–12. Page 12/21 Page 12/21 Page 12/21 34. Li Y, Du X, Liu C, Wen Z, Du J. Reciprocal Regulation between Resting Microglial Dynamics and Neuronal Activity In
Vivo. Dev Cell. 2012;23:1189–202. 35. Cserép C, Pósfai B, Lénárt N, Fekete R, László ZI, Lele Z, et al. Microglia monitor and protect neuronal function
through specialized somatic purinergic junctions. Science. 2020;367:528–37. 35. Cserép C, Pósfai B, Lénárt N, Fekete R, László ZI, Lele Z, et al. Microglia monitor and protect neuronal function
through specialized somatic purinergic junctions. Science. 2020;367:528–37. 36. Shi T-JS, Tandrup T, Bergman E, Xu Z-QD, Ulfhake B, Hökfelt T. Effect of peripheral nerve injury on dorsal root
ganglion neurons in the C57 BL/6J mouse: marked changes both in cell numbers and neuropeptide expression. Neuroscience. 2001;105:249–63. 36. Shi T-JS, Tandrup T, Bergman E, Xu Z-QD, Ulfhake B, Hökfelt T. Effect of peripheral nerve injury on dorsal root
ganglion neurons in the C57 BL/6J mouse: marked changes both in cell numbers and neuropeptide expression. Neuroscience. 2001;105:249–63. 37. Whiteside GT, Munglani R. Cell death in the superficial dorsal horn in a model of neuropathic pain. J Neurosci Res. 2001;64:168–73. 37. Whiteside GT, Munglani R. Cell death in the superficial dorsal horn in a model of neuropathic pain. J Neurosci Res. 2001;64:168–73. 38. Schaeffer V, Meyer L, Patte‐mensah C, Eckert A, Mensah‐nyagan AG. Sciatic nerve injury induces apoptosis of
dorsal root ganglion satellite glial cells and selectively modifies neurosteroidogenesis in sensory neurons. Glia. 2010;58:169–80. 39. Groves MJ, Christopherson T, Giometto B, Scaravilli F. Axotomy-induced apoptosis in adult rat primary sensory
neurons. J Neurocytol. 1997;26:615–24. 40. Lyu C, Lyu G-W, Martinez A, Shi T-JS. Effect of nerve injury on the number of dorsal root ganglion neurons and
autotomy behavior in adult Bax-deficient mice. J Pain Res. 2017;10:2079–87. 40. Lyu C, Lyu G-W, Martinez A, Shi T-JS. Effect of nerve injury on the number of dorsal root ganglion neurons and
autotomy behavior in adult Bax-deficient mice. J Pain Res. 2017;10:2079–87. 41. Coggeshall RE, Lekan HA, Doubell TP, Allchorne A, Woolf CJ. Central changes in primary afferent fibers following
peripheral nerve lesions. Neuroscience. 1997;77:1115–22. 41. Coggeshall RE, Lekan HA, Doubell TP, Allchorne A, Woolf CJ. Central changes in primary afferent fibers following
peripheral nerve lesions. Neuroscience. 1997;77:1115–22. Tables Table 1 Antibodies used in the present study Page 13/21 Primary antibody
Catalog No. Host
Clonality
Concentration
Manufacturer
RRID
Anti-activating
transcription factor
3 (ATF3)
HPA001562
Rabbit
Polyclonal
1:1000
Atlas Antibodies,
Stockholm,
Sweden
AB_1078233
Anti-
bromodeoxyuridine
(BrdU)
ab6326
Rat
Monoclonal
1:1000
Abcam,
Cambridge, UK
AB_305426
Anti-ionized
calcium binding
adaptor molecule 1
(Iba1)
ab5076
Goat
Polyclonal
1:5000
Abcam,
Cambridge, UK
AB_2224402
Anti-product gene
protein 9.5
(PGP9.5)
GP14104
Guinea
pig
Polyclonal
antiserum
1:5000
Neuromics,
Edina, MN
AB_2210625
Anti-caspase 3
NB600-
1235
Rabbit
Polyclonal
1:500
Novus
Biologicals,
Littleton, CO
AB_2069897
Anti-glutamine
synthetase
G2781
Rabbit
Polyclonal
1:10000
Merck,
Darmstadt,
Germany
AB_259853
Anti-mouse cluster
of differentiation
206 (CD206)
AF2535
Goat
Polyclonal
1:1000
R&D Systems,
Minneapolis, MN
AB_2063012
Anti-mouse cluster
of differentiation
86 (CD86)
553689
Rat
Monoclonal
1:1000
R&D Systems,
Minneapolis, MN
AB_394991
Secondary
antibody
Catalog No. Host
Clonality
Concentration
Manufacturer
RRID
Anti-rat IgG-
conjugated
to Alexa Fluor 488
ab150153
Donkey
Polyclonal
1:800
Abcam,
Cambridge, UK
AB_2737355
Anti-rabbit IgG-
conjugated
to Alexa Fluor 488
ab150065
Donkey
Polyclonal
1:800
Abcam,
Cambridge, UK
AB_2860569
Anti-rabbit IgG-
conjugated
to Alexa Fluor 555
ab150066
Donkey
Polyclonal
1:800
Abcam,
Cambridge, UK
Anti-rabbit IgG-
conjugated
to Alexa Fluor 647
ab150067
Donkey
Polyclonal
1:800
Abcam,
Cambridge, UK Anti-goat IgG-
conjugated
to Alexa Fluor 647
A-21447
Donkey
Polyclonal
1:800
Thermo Fisher
Scientific,
Waltham, MA
AB_2535864
Anti-Guinea pig
IgG-conjugated
to Alexa Fluor 488
706-545-
148
Donkey
Polyclonal
1:800
Jackson
ImmunoResearch
Labs, West
Grove, PA
AB_2340472
Anti-Guinea pig
IgG-conjugated
to Alexa Fluor 647
706-605-
148
Donkey
Polyclonal
1:800
Jackson
ImmunoResearch
Labs, West
Grove, PA
AB_2340476
Anti-rabbit IgG-
conjugated
to biotin
BA-1000
Goat
Polyclonal
1:200
Vector
Laboratories,
Burlingame, CA
AB_2313606
RRID research resource identifier
The English in this document has been checked by at least two professional editors, both native speakers of English. For a certificate, please see:
http://www.textcheck.com/certificate/5IV0qL Anti-goat IgG-
conjugated
to Alexa Fluor 647
A-21447
Donkey
Polyclonal
1:800
Thermo Fisher
Scientific,
Waltham, MA
AB_2535864
Anti-Guinea pig
IgG-conjugated
to Alexa Fluor 488
706-545-
148
Donkey
Polyclonal
1:800
Jackson
ImmunoResearch
Labs, West
Grove, PA
AB_2340472
Anti-Guinea pig
IgG-conjugated
to Alexa Fluor 647
706-605-
148
Donkey
Polyclonal
1:800
Jackson
ImmunoResearch
Labs, West
Grove, PA
AB_2340476
Anti-rabbit IgG-
conjugated
to biotin
BA-1000
Goat
Polyclonal
1:200
Vector
Laboratories,
Burlingame, CA
AB_2313606 Figures Page 15/21 Figure 1
Ganglionic macrophages proliferate after nerve injury. ATF3-positive cells (green) in the contra
ipsilateral (ipsi) sides of the maxillary nerve region of the trigeminal ganglia on day 1 after inf
and the number of ATF3-positive cells (n = 4–6/time points) (b). Iba1-positive cells (red) on da
(c) and the number of Iba1-positive cells (n = 4–6/time points) (d). BrdU (green)- and Iba1 (red
multiple staining showing co-localization (arrowhead) of BrdU signals (green) with nucleus (b
(red) (f) on day 5 after a nerve ligation, and the number of BrdU- and Iba1-positive cells (n = 4–
list of abbreviations. Scale bars are indicated. Data are represented as mean (S.D.), and differ
two-way ANOVA with Tukey-Kramer test (b, d, g). Figure 1 Ganglionic macrophages proliferate after nerve injury. ATF3-positive cells (green) in the contralateral (contra) and
ipsilateral (ipsi) sides of the maxillary nerve region of the trigeminal ganglia on day 1 after infraorbital nerve ligation (a)
and the number of ATF3-positive cells (n = 4–6/time points) (b). Iba1-positive cells (red) on day 7 after nerve ligation
(c) and the number of Iba1-positive cells (n = 4–6/time points) (d). BrdU (green)- and Iba1 (red)-positive cells (e),
multiple staining showing co-localization (arrowhead) of BrdU signals (green) with nucleus (blue) of Iba1-posive cells
(red) (f) on day 5 after a nerve ligation, and the number of BrdU- and Iba1-positive cells (n = 4–6/time points) (g). See
list of abbreviations. Scale bars are indicated. Data are represented as mean (S.D.), and differences were detected using
two-way ANOVA with Tukey-Kramer test (b, d, g). Page 16/21 Figure 2
BMD macrophages increase in sensory ganglia after nerve injury. GFP-BMT mice after wearing a lead cap and was
irradiated, and wild-type mice (a). GFP (green)- and Iba1 (red)-positive cells (b) and multiple staining showing co-
localization (arrowhead) of GFP signals (green) with Iba1-posive cells (red) (c) in the contralateral (contra) and
ipsilateral (ipsi) sides of the maxillary nerve region of the trigeminal ganglia on day 7 after nerve transection. The
number of GFP-positive cells (n = 7/group) (d), GFP- and Iba1-positive cells (n = 7/group) (e), and GFP-negative and
Iba1-positive cells or GFP- and Iba1-positive cells (n = 7/group) (f) on day 7 after nerve transection. See list of Figure 2 Figure 2 BMD macrophages increase in sensory ganglia after nerve injury. GFP-BMT mice after wearing a lead cap and was
irradiated, and wild-type mice (a). GFP (green)- and Iba1 (red)-positive cells (b) and multiple staining showing co-
localization (arrowhead) of GFP signals (green) with Iba1-posive cells (red) (c) in the contralateral (contra) and
ipsilateral (ipsi) sides of the maxillary nerve region of the trigeminal ganglia on day 7 after nerve transection. The
number of GFP-positive cells (n = 7/group) (d), GFP- and Iba1-positive cells (n = 7/group) (e), and GFP-negative and
Iba1-positive cells or GFP- and Iba1-positive cells (n = 7/group) (f) on day 7 after nerve transection. See list of
abbreviations. Scale bars are indicated. Data are represented as mean (S.D.), and differences were detected using
Student’s t-test (d) and Student’s t-test with Welch’s correction (e, f). Page 17/21 Page 17/21 Page 17/21 Figure 3
The volume of ganglionic macrophages enlarges after nerve injury. Iba1-positive cells (red) in the contralatera
and ipsilateral (ipsi) sides of the maxillary nerve region of the trigeminal ganglia on day 7 after infraorbital ne
ligation (a) and the cell areas of Iba1-positive cells (n = 4–6/time points) (b). Z-stack images showing Iba1-po
cells (red) around ATF3-positive cells (green) (c), three-dimensional images showing Iba1-positive cells (d), an
volume of Iba1-positive cells (n = 10 cells/group from 3 mice) (e) on day 7 after nerve ligation. See list of abb
Scale bars are indicated. Data are represented as mean (S.D.), and differences were detected using two-way A
with Tukey-Kramer test (b) and Student’s t-test with Welch’s correction (e). Figure 3
The volume of ganglionic macrophages enlarges after nerve injury. Iba1-positive cells (red) in the contralat
and ipsilateral (ipsi) sides of the maxillary nerve region of the trigeminal ganglia on day 7 after infraorbital
ligation (a) and the cell areas of Iba1-positive cells (n = 4–6/time points) (b). Z-stack images showing Iba1
cells (red) around ATF3-positive cells (green) (c), three-dimensional images showing Iba1-positive cells (d), Figure 3 The volume of ganglionic macrophages enlarges after nerve injury. Iba1-positive cells (red) in the contralateral (contra)
and ipsilateral (ipsi) sides of the maxillary nerve region of the trigeminal ganglia on day 7 after infraorbital nerve
ligation (a) and the cell areas of Iba1-positive cells (n = 4–6/time points) (b). Z-stack images showing Iba1-positive
cells (red) around ATF3-positive cells (green) (c), three-dimensional images showing Iba1-positive cells (d), and the cell
volume of Iba1-positive cells (n = 10 cells/group from 3 mice) (e) on day 7 after nerve ligation. See list of abbreviations. Scale bars are indicated. Data are represented as mean (S.D.), and differences were detected using two-way ANOVA
with Tukey-Kramer test (b) and Student’s t-test with Welch’s correction (e). Page 18/21 Page 18/21 Figure 4 Figure 4
The contact area of ganglionic macrophages to primary sensory neurons are expanded after nerve injury. ATF3
Iba1 (red)-, and PGP9.5 (blue)-positive cells in ipsilateral (ipsi) sides of the maxillary nerve region of the trigemin
ganglia on day 1 and 7 after infraorbital nerve ligation (a) and a percentage of contact-like structure (CLS) betw
ATF3-positive or negative neurons and Iba1-positive cells (n = 4 or 5/group) (b). Multiple staining (c) and Z-stac
images (d) showing Iba1-positive cells (green), Nissl-positive neurons (blue), and glutamine synthetase (GS)-po
cells (red) in the contralateral (contra) and ipsilateral sides on day 7 after nerve ligation. The contact sites betw
Iba1-positive cells and neurons are indicated by arrowhead. Three-dimensional images showing contact area of
positive cells and Nissl-positive neurons (e). Blue indicates Nissl-positive neurons, and yellow indicates areas w
Iba1-positive cells and neurons are in contact. Surface area of Iba1-positive or GS-positive cells in contact with Figure 4 The contact area of ganglionic macrophages to primary sensory neurons are expanded after nerve injury. ATF3 (green)-,
Iba1 (red)-, and PGP9.5 (blue)-positive cells in ipsilateral (ipsi) sides of the maxillary nerve region of the trigeminal
ganglia on day 1 and 7 after infraorbital nerve ligation (a) and a percentage of contact-like structure (CLS) between
ATF3-positive or negative neurons and Iba1-positive cells (n = 4 or 5/group) (b). Multiple staining (c) and Z-stack
images (d) showing Iba1-positive cells (green), Nissl-positive neurons (blue), and glutamine synthetase (GS)-positive
cells (red) in the contralateral (contra) and ipsilateral sides on day 7 after nerve ligation. The contact sites between
Iba1-positive cells and neurons are indicated by arrowhead. Figure 3 Three-dimensional images showing contact area of Iba1-
positive cells and Nissl-positive neurons (e). Blue indicates Nissl-positive neurons, and yellow indicates areas where
Iba1-positive cells and neurons are in contact. Surface area of Iba1-positive or GS-positive cells in contact with The contact area of ganglionic macrophages to primary sensory neurons are expanded after nerve injury. ATF3 (green)-,
Iba1 (red)-, and PGP9.5 (blue)-positive cells in ipsilateral (ipsi) sides of the maxillary nerve region of the trigeminal
ganglia on day 1 and 7 after infraorbital nerve ligation (a) and a percentage of contact-like structure (CLS) between
ATF3-positive or negative neurons and Iba1-positive cells (n = 4 or 5/group) (b). Multiple staining (c) and Z-stack
images (d) showing Iba1-positive cells (green), Nissl-positive neurons (blue), and glutamine synthetase (GS)-positive
cells (red) in the contralateral (contra) and ipsilateral sides on day 7 after nerve ligation. The contact sites between
Iba1-positive cells and neurons are indicated by arrowhead. Three-dimensional images showing contact area of Iba1-
positive cells and Nissl-positive neurons (e). Blue indicates Nissl-positive neurons, and yellow indicates areas where
Iba1-positive cells and neurons are in contact. Surface area of Iba1-positive or GS-positive cells in contact with Page 19/21 Page 19/21 individual Nissl-positive neurons (f) and a percentage of these areas averaged together (averaging 10 cells/ipsi from 3
mice) (g) on day 7 after nerve ligation. See list of abbreviations. Scale bars are indicated. Figure 5 Figure 5
Ganglionic macrophages in sensory ganglia after nerve injury are categorized to “tissue repair”. CD206-positive cells
(green) (a), multiple staining showing co-localization (arrowhead) of CD206 signals (green) with Iba1-posive cells (red)
(b), and the number of CD206- and Iba1-positive cells (n = 4/group) (c) in contralateral (contra) and ipsilateral (ipsi)
sides of the maxillary nerve region of the trigeminal ganglia on day 7 after infraorbital nerve ligation. CD86-positive
cells (green) (d), multiple staining showing co-localization (arrowhead) of CD86 signals (green) with Iba1-posive cells
(red) (e), and the number of CD86- and Iba1-positive cells (n = 4/group) (f) on day 7 after infraorbital nerve ligation. Caspase-3-positive cells (brown, arrowhead) on week 1 and 4 after nerve ligation (g) and the number of caspase-3-
positive cells (n = 4/time point) (h). Caspase-3 (green)- and PGP9.5 (red)-positive cells (i) and the percentage of Figure 5 Ganglionic macrophages in sensory ganglia after nerve injury are categorized to “tissue repair”. caspase-3-positive neurons (n = 129 cells/ipsi from 4 mice) (j) on day 7 after nerve ligation. See list of abbreviations.
Scale bars are indicated. Data are represented as mean (S.D.), and differences were detected using Student’s t-test (c),
Student’s t-test with Welch’s correction (f), and one-way ANOVA with Tukey-Kramer test (h). Figure 3 CD206-positive cells
(green) (a), multiple staining showing co-localization (arrowhead) of CD206 signals (green) with Iba1-posive cells (red)
(b), and the number of CD206- and Iba1-positive cells (n = 4/group) (c) in contralateral (contra) and ipsilateral (ipsi)
sides of the maxillary nerve region of the trigeminal ganglia on day 7 after infraorbital nerve ligation. CD86-positive
cells (green) (d), multiple staining showing co-localization (arrowhead) of CD86 signals (green) with Iba1-posive cells
(red) (e), and the number of CD86- and Iba1-positive cells (n = 4/group) (f) on day 7 after infraorbital nerve ligation. Caspase-3-positive cells (brown, arrowhead) on week 1 and 4 after nerve ligation (g) and the number of caspase-3-
positive cells (n = 4/time point) (h). Caspase-3 (green)- and PGP9.5 (red)-positive cells (i) and the percentage of Page 20/21 Page 20/21 caspase-3-positive neurons (n = 129 cells/ipsi from 4 mice) (j) on day 7 after nerve ligation. See list of abbreviations. Scale bars are indicated. Data are represented as mean (S.D.), and differences were detected using Student’s t-test (c),
Student’s t-test with Welch’s correction (f), and one-way ANOVA with Tukey-Kramer test (h). caspase-3-positive neurons (n = 129 cells/ipsi from 4 mice) (j) on day 7 after nerve ligation. See list of abbreviations. Scale bars are indicated. Data are represented as mean (S.D.), and differences were detected using Student’s t-test (c),
Student’s t-test with Welch’s correction (f), and one-way ANOVA with Tukey-Kramer test (h). caspase-3-positive neurons (n = 129 cells/ipsi from 4 mice) (j) on day 7 after nerve ligation. See list of abbreviations. Scale bars are indicated. Data are represented as mean (S.D.), and differences were detected using Student’s t-test (c),
Student’s t-test with Welch’s correction (f), and one-way ANOVA with Tukey-Kramer test (h). Page 21/21
|
https://openalex.org/W2048171954
|
https://www.scielo.br/j/rep/a/rcyNzvhkRKfsk6Hrn8sYLJQ/?lang=pt&format=pdf
|
Portuguese
| null |
O centenário de Ignácio Rangel
|
Brazilian Journal of Political Economy
| 2,014
|
cc-by
| 11,560
|
* Doutor em Economia pela Universidade Federal de Pernambuco (UFPE). Professor adjunto aposentado
da Universidade Federal de Santa Maria (UFSM) e professor do curso de Ciências Econômicas do Centro
Universitário Franciscano, em Santa Maria (RS). E-mail: pereirajm@uol.com.br. Artigo baseado em
capítulo do livro Centenário de Ignácio Rangel, edição conjunta da Universidade Federal do Maranhão
e Conselho Regional de Economia do Maranhão, 2014. Revista de Economia Política, vol. 34, nº 4 (137), pp. 544-564, outubro-dezembro/2014 Revista de Economia Política, vol. 34, nº 4 (137), pp. 544-564, outubro-dezembro/2014 JOSÉ MARIA DIAS PEREIRA* JOSÉ MARIA DIAS PEREIRA* In February 2014, celebrating the centenary of Ignácio Rangel — one of the
most original thinkers of the Brazilian economy. This article aims to summarize
the main points of his work, considering that his theoretical contribution is poorly
known of the new generations. It was one of the first to associate the internal cycles
of our economy to the cycles of the world economy through the thesis of duality. Inflation was part of the “syndrome of the recession.” Out of recession, there should
be transfer of resources from activities with idle capacity for latecomers sectors, in
case public services. Keywords: Ignácio Rangel; economic cycles; inflation and recession. JEL Classification: E31; E32; E61. Keywords: Ignácio Rangel; economic cycles; inflation and recession. JEL Classification: E31; E32; E61. 1 “Economista Ignácio Rangel morre no Rio”, Folha de São Paulo, 5/3/1994. Keywords: Ignácio Rangel; economic cycles; inflation and recession.
JEL Classification: E31; E32; E61. INTRODUÇÃO O economista Ignácio de Mourão Rangel nasceu no Maranhão, em 20 de fe-
vereiro de 1914, e faleceu, no Rio de Janeiro, em quatro de março de 1994. Por-
tanto, em 2014, comemora-se o centenário do nascimento do grande mestre. Fale-
cido aos 80 anos, Rangel foi, segundo o próprio jornal que noticiou o seu
falecimento, “um dos mais importantes analistas econômicos do país” 1. Mas Ran-
gel foi muito mais do que isso. Formado em Direito, aprendeu Economia na práti-
ca, participando da assessoria econômica do 2º governo Vargas, onde colaborou
para a criação da Petrobras e Eletrobras. Em 1956, foi trabalhar no BNDES, par-
ticipando da elaboração do Plano de Metas do governo de Juscelino Kubitschek. 544 Brazilian Journal of Political Economy 34 (4), 2014 Foi também um entusiasta da industrialização do país, fazendo parte, desde o início,
do Instituto Superior de Estudos Brasileiros (ISEB) — núcleo acadêmico que abri-
gava os teóricos do nacional-desenvolvimentismo. Rangel colaborou com univer-
sidades, como a Unicamp e UFMG, e participou de órgãos de classe, tendo sido
presidente de Conselho Regional de Economia do Rio de Janeiro. Foi também um entusiasta da industrialização do país, fazendo parte, desde o início,
do Instituto Superior de Estudos Brasileiros (ISEB) — núcleo acadêmico que abri-
gava os teóricos do nacional-desenvolvimentismo. Rangel colaborou com univer-
sidades, como a Unicamp e UFMG, e participou de órgãos de classe, tendo sido
presidente de Conselho Regional de Economia do Rio de Janeiro. Rangel se destacou ainda como um teórico extremamente original da Econo-
mia, expondo suas ideias através de vários livros e artigos em periódicos e jornais. Um desses periódicos foi justamente a Revista de Economia Política, fundada em
1981 por um grupo de economistas que rejeitava a visão dominante da economia,
como ciência positiva e neutra, e defendia um retorno às suas origens clássicas,
como “Economia Política”. Rangel destacou-se, desde o primeiro número, como
um dos mais frequentes colaboradores e mereceu a honra de ver o seu nome como
patrono da revista, ao lado de Caio Prado Jr. e o de Celso Furtado. Tomei conhecimento da teoria da inflação de I. Rangel, por ocasião da elabo-
ração da minha dissertação de mestrado (Pereira, 1984), defendida na Universida-
de Federal do Rio Grande do Sul (UFRGS), ao pesquisar a influência da alimenta-
ção na inflação brasileira nos anos 1970. INTRODUÇÃO Na época (início dos anos 1980), o
regime militar já “fazia água” e o debate sobre as causas da inflação assumia um
contorno claramente ideológico entre estruturalistas (esquerda) e monetaristas (di-
reita). O livro A inflação brasileira, de I. Rangel, que havia sido recentemente (1978)
reeditado, abria uma nova perspectiva de interpretação da inflação, com ênfase na
estrutura concentrada de comercialização de produtos agrícolas. Em 1982, quando
Rangel deu uma palestra na UFRGS, por ocasião do lançamento do seu livro Ciclo,
tecnologia e crescimento, conheci-o pessoalmente. Cerca de uma década depois, quando fazia meu doutoramento, na Universi-
dade Federal de Pernambuco (UFPE), no Recife, Rangel fez uma espécie de “mini-
conferência” para os estudantes da Pós-graduação. Foi daí que surgiu a ideia de
escrever uma “comunicação” sobre os 30 anos da 1ª edição do seu livro A inflação
brasileira, publicada pela Revista de Economia Política (v. 3, n. 13, jul.-set./1993). Meu derradeiro encontro com I. Rangel se deu cerca de um ano antes de seu fale-
cimento, numa palestra na Universidade Federal de Santa Maria (UFSM). Com a
saúde debilitada, Rangel falava com dificuldade, mas com o entusiasmo de sempre. Deixou um legado de valor inestimável, mas infelizmente pouco reconhecido. Além
do propósito de homenagear o grande mestre pelo centenário do seu nascimento,
o autor deste artigo pretendeu também divulgar suas principais ideias entre as
novas gerações de economistas. Revista de Economia Política 34 (4), 2014 • pp. 544-564 ESQUERDA OU DIREITA? Dentre os traços da personalidade marcante de Ignácio Rangel, o que mais
fascinava era a sua capacidade crítica. Seria injusto não classificá-lo como um Revista de Economia Política 34 (4), 2014 • pp. 544-564 545 homem de esquerda2, mas, em determinado momento, foi rejeitado pela esquerda
como sendo de direita. Isso é especialmente verdadeiro quando, nos anos 1950 e
1960, discordou da tese da esquerda de que a reforma agrária era uma “précondi-
ção” para o desenvolvimento. Rangel argumentava, na época, que não havia con-
dições objetivas para a realização de uma reforma agrária no Brasil, mas que isso,
em absoluto, inviabilizaria a retomada do crescimento econômico. Mesmo após o
encerramento do ciclo de crescimento dos anos 1970, quando passou a defender a
reforma agrária, continuou a ser alvo da incompreensão da esquerda3. Sua visão sobre desenvolvimento ia além das ideologias de esquerda e direita. Na trilha de Keynes e Kalecki, colocava o investimento como condição fundamen-
tal para o desenvolvimento. Considerando a condição do Brasil como país perifé-
rico, a princípio, propôs que o Estado bancasse o financiamento ao investimento. Com a crise das finanças públicas dos anos 1960, sugeriu ao ministro da Fazenda
do primeiro governo militar a criação da correção monetária como estratégia al-
ternativa para manter os investimentos. Posteriormente — sendo este mais um
exemplo do pragmatismo de Rangel —, em face das distorções causadas pela cor-
reção monetária, no final dos anos 1970, sugeriu a criação de uma nova lei de
concessão de serviços públicos. Através dela, o setor privado passaria a encarregar-
-se, de forma crescente, pelos investimentos públicos de infraestrutura. Essa sugestão
não deixa de ser surpreendente, considerando que Rangel teve um papel importan-
te na criação de grandes empresas estatais brasileiras, como a Petrobras e a Eletro-
bras. Nem em sonhos Rangel poderia imaginar que, após o seu falecimento (1994),
a privatização de empresas estatais iria tão longe. No que se refere à inflação, foi a mesma coisa. Advogado por formação, o
único curso formal de economia que Rangel frequentou foi, nos anos 1950, na
Comissão Econômica para a América Latina e Caribe (CEPAL), templo do estru-
turalismo latino-americano, no Chile. Nem por isso, Rangel poupou o estrutura-
lismo na sua análise das causas da inflação brasileira. 3 Numa crônica publicada no Jornal do Brasil, em 18/9/1979, reclamava da assessoria econômica de
Miguel Arraes que continuava criticando-o por ser contra a reforma agrária quando, há muito, havia
mudado de opinião. Escreveu ele: “Em matéria de assessoramento econômico, nossa esquerda é, em geral,
uma catástrofe [...]. Ora, Miguel Arraes não constitui exceção. Sua assessoria não aprendeu nada, nem
esqueceu nada, nestes três lustros [...] Só faltaria que, agora, em 1979, quando eu estou sustentando que
isso (tese contrária à reforma agrária como précondição para o desenvolvimento) não é mais verdade,
eles levantem a bandeira do meu velho livro (refere-se ao livro A questão agrária brasileira), para afirmar
aquelas verdades tornadas caducas. Não seria essa a primeira vez” (Rangel, 1982b, p. 156). 2 Segundo Maria da Conceição Tavares, Rangel “foi o grande patrono dos chamados economistas de
esquerda” (Folha de São Paulo, 5/3/1994). 4 Em crônica já citada, Rangel desabafou: “Nada me obriga a escolher líder algum, porque, afinal, não
sou apenas um cidadão, mas também um economista. Um economista de esquerda (grifo do autor) [...]
que continuou, infatigavelmente, a trabalhar seus esquemas, contra o vento e a maré, mas que tem a
consciência de estar vencendo. Que viu chegar o momento de ver suas ideias — aquelas pelas quais se
bateu, solitariamente, por tantos anos, expondo-se à chacota dos doutores — serem postas em circulação,
sem referência ao seu nome”. Ibid em, nota 2, p. 155. Numa possível identificação aos “doutores” a
que Rangel se referia, Bresser-Pereira (1993) lembra-se de um seminário na USP organizado por Delfim
Netto, em 1964, do qual participou, em que as teses de Rangel sobre inflação foram criticadas.
Paradoxalmente, ao assumir o Ministério da Fazenda, em 1967, Delfim colocou em prática uma política
econômica consistente com a tese de Rangel de que a inflação brasileira, na época, era fundamentalmente
de custos. ESQUERDA OU DIREITA? No clássico A inflação bra-
sileira, publicado no limiar do regime militar (1963), Rangel ironizava os “nossos
ortodoxos estruturalistas” por confundirem a estrutura concentrada de interme-
diação — causa principal da alta do preço dos alimentos — com inelasticidade da
oferta agrícola. Igualmente, não poupava os “nossos ortodoxos monetaristas” por
não se darem conta que a moeda é passiva, ou seja, é o aumento de preços que Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 546 leva as autoridades monetárias a aumentarem a quantidade de moeda e não o
contrário. Não é difícil imaginar que, não se filiando a nenhuma corrente dominante de
economistas de esquerda ou de direita, Rangel fosse, com raras exceções, solene-
mente ignorado por ambos os lados. Em certo momento, deixou transparecer al-
guma mágoa em relação a isso4. O que poucos perceberam é que Rangel tinha
maior autonomia de voo. Ambicionava construir uma teoria dos ciclos da economia
brasileira que não fosse simplesmente reflexo das vicissitudes das economias do
centro capitalista desenvolvido. Para chegar até ela, baseou-se nas teorias dos ciclos
longos (50 anos) do russo N. Kondratieff e dos ciclos médios (10 anos) do francês
C. Juglar. Do ponto de vista da teoria econômica, foi buscar subsídios em Marx,
em Keynes e em Schumpeter5, dentre outros. Surgia, assim, a tese da dualidade da
economia brasileira. CICLOS E DUALIDADE A tese da dualidade, segundo Bielschowsky (1996), é o principal elemento
organizador do pensamento de Ignácio Rangel. Ela resulta de uma adaptação do
materialismo histórico marxista ao caso específico da economia brasileira, porém,
ao invés de uma sucessão de modos de produção, subdividiu o conceito marxista
de “relações de produção” em relações internas e relações externas6. Por seu turno,
cada uma dessas relações teria também um lado interno e outro externo. Quando
são preenchidas as condições para a passagem a um estágio superior — quando as 4 Em crônica já citada, Rangel desabafou: “Nada me obriga a escolher líder algum, porque, afinal, não
sou apenas um cidadão, mas também um economista. Um economista de esquerda (grifo do autor) [...]
que continuou, infatigavelmente, a trabalhar seus esquemas, contra o vento e a maré, mas que tem a
consciência de estar vencendo. Que viu chegar o momento de ver suas ideias — aquelas pelas quais se
bateu, solitariamente, por tantos anos, expondo-se à chacota dos doutores — serem postas em circulação,
sem referência ao seu nome”. Ibid em, nota 2, p. 155. Numa possível identificação aos “doutores” a
que Rangel se referia, Bresser-Pereira (1993) lembra-se de um seminário na USP organizado por Delfim
Netto, em 1964, do qual participou, em que as teses de Rangel sobre inflação foram criticadas. Paradoxalmente, ao assumir o Ministério da Fazenda, em 1967, Delfim colocou em prática uma política
econômica consistente com a tese de Rangel de que a inflação brasileira, na época, era fundamentalmente
de custos. 5 O modelo do “ciclo real de negócios” (Sachs-Larrain, 2000) — baseado na tese de Joseph Schumpeter
de que o capitalismo é caracterizado por ondas de “destruição criadora” , provocadas por choques
aleatórios de tecnologia — teve grande influência no pensamento de Rangel, como se pode concluir da
famosa frase de Schumpeter de que “os ciclos não são, como as amígdalas, coisas que podem ser
separáveis do corpo [...]”, citada por Rangel (1985a). 6 Segundo Bresser-Pereira (1993, p. 102): “O método utilizado por Rangel para analisar a economia
brasileira sempre foi essencialmente histórico. Sua origem marxista é evidente. Mas Rangel usa Marx
com absoluta liberdade [...] Rangel não se deixa levar por uma postura ideológica romântica em relação
à burguesia, não aplica o método mecanicamente através de simples transposição das fases históricas
ocorridas na Europa que Marx estudou, não se esquece jamais das especificidades brasileiras”. CICLOS E DUALIDADE 547 Revista de Economia Política 34 (4), 2014 • pp. 544-564 forças produtivas da sociedade crescem, entrando em conflito com as relações de
produção existentes — surge uma “nova dualidade”, porém apenas um dos seus
lados muda, permanecendo o outro com a mesma estrutura. Esta é a “1ª lei da
dualidade brasileira” 7. Posteriormente, Rangel incorporou o conceito de ciclo longo à tese da duali-
dade. Nesse sentido, resgatou a teoria dos ciclos longos do economista russo Ni-
kolai Kondratieff8, desenvolvida por volta dos anos 1920. Os ciclos de Kondratie-
ff tem duração aproximada de meio século, dividido em um quarto de século na
fase “a”, ou ascendente, e outro quarto na fase “b” ou descendente. O Brasil se
insere nos ciclos longos da economia mundial da seguinte forma: na fase ascenden-
te, é favorecido pelo aumento de suas exportações; na fase descendente (crise),
devido à escassez de divisas, se dedica a substituir importações. As fases “a” do
ciclo de Kondratieff foram 1790-1815, 1847-1873 e 1896-1920, enquanto as fases
“b” compreenderam os períodos 1815-1847, 1873-1896 e 1920-1948. Rangel inclui
um 4º ciclo de Kondratieff, sendo 1948-1973 a fase “a” e após 1973 (1º choque do
petróleo) teria começado a fase “b” 9. O que diferencia o caso brasileiro das demais formações históricas clássicas,
segundo Ignácio Rangel, é que existe uma interação entre o desenvolvimento das
forças produtivas e das relações de produção internas e externas e, por causa da
sua dependência em relação às economias centrais, estas últimas são determinantes
em última instância do desenvolvimento das primeiras. Para explicar o desenvol-
vimento econômico, Rangel divide a economia brasileira em três setores: o primei-
ro, e mais atrasado, formado pela agricultura feudal ou pré-capitalista (ou de sub-
sistência); o segundo, pela economia de mercado capitalista (todas as demais
atividades) e, o terceiro, pelo comércio exterior. A transformação da economia
seria sempre resultado da ação que o segundo setor (o capitalista) exerce sobre o
primeiro, a partir de estímulos oriundos do terceiro setor. A contrapartida política da tese da dualidade estaria na base da formação do
Estado brasileiro. A mudança de um modo de produção para outro está associado a
um “pacto de poder” que resulta da aliança de apenas duas classes dirigentes, uma
representando o segmento interno e outra o segmento externo. 8 Segundo Rangel (1982, p. 17), “Kondratieff tornou-se um profeta maldito dos dois lados da Cortina
de Ferro”. Na ex-União Soviética, foi destituído de seus cargos, preso e deportado para a Sibéria. No
Ocidente, se não fosse Schumpeter, que usou o seu nome para denominar os ciclos longos, teria sido um
ilustre desconhecido. Rangel atribui esse descaso ao fato de que a sua teoria era desfavorável tanto ao
capitalismo, por prever que, de tempos em tempos, a prosperidade acabaria (entraria na fase “b”);
quanto ao socialismo, por admitir que, após cada crise, o capitalismo não se enfraqueceria — como
previra Lênin — mas se fortaleceria (entraria novamente na fase “a”). 9 Ver, a esse respeito, o texto “O Brasil na fase “b” do 4º Kondratieff” incluído na coletânea Ciclo,
tecnologia e crescimento (Rangel, 1982). 7 Cf. Rangel (1981, p. 12). CICLOS E DUALIDADE A transição, de uma
dualidade para outra, se faz por cooptação, isto é, “pela exclusão, pelo próprio gru-
po dirigente, dos elementos mais arcaicos, e sua substituição por outros, representa- 9 Ver, a esse respeito, o texto “O Brasil na fase “b” do 4º Kondratieff” incluído na coletânea Ciclo,
tecnologia e crescimento (Rangel, 1982). Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 548 tivos das novas forças sociais em ascensão” 10. A história das dualidades brasileiras,
segundo a cronologia de Ignácio Rangel, é mostrada nos diagramas a seguir11. A primeira dualidade brasileira, assim como as que vieram depois, formou-se
na fase “b” do ciclo longo do 1º Kondratieff. O polo externo definiu-se com a
substituição do capital mercantil português pelo nascente capitalismo mercantil
brasileiro. Surgia nesse momento a classe dos comerciantes (principalmente expor-
tadores-importadores), que seria uma das classes dirigentes do Estado no papel de
sócio menor. Esses comerciantes, predominantemente estrangeiros, ainda estavam
politicamente despreparados para o exercício do poder, mas eram uma espécie de
“correia de transmissão” através da qual o capital industrial do centro dinâmico
impulsionava todo o sistema de acordo com os seus interesses. A primeira dualidade brasileira, assim como as que vieram depois, formou-se
na fase “b” do ciclo longo do 1º Kondratieff. O polo externo definiu-se com a
substituição do capital mercantil português pelo nascente capitalismo mercantil
brasileiro. Surgia nesse momento a classe dos comerciantes (principalmente expor-
tadores-importadores), que seria uma das classes dirigentes do Estado no papel de
sócio menor. Esses comerciantes, predominantemente estrangeiros, ainda estavam
politicamente despreparados para o exercício do poder, mas eram uma espécie de
“correia de transmissão” através da qual o capital industrial do centro dinâmico
impulsionava todo o sistema de acordo com os seus interesses. No polo interno, a sociedade estruturava-se em torno da fazenda de escravos. Emergia dela o sócio maior da primeira dualidade, ou seja, a classe dos barões
senhores de escravos. Dois acontecimentos — a independência (1822) e a abdi-
cação de Dom Pedro I (1831) — são marcos históricos das mudanças efetivadas. No lado externo, havia toda uma pressão, sobretudo da Inglaterra, para que o
escravismo fosse substituído pelo latifúndio feudal. No lado interno, o comércio
de escravos abastecia regularmente não só a lavoura como também uma relativa
diversificação da fazenda de escravos. 11 A análise que segue, assim como a elaboração dos diagramas, está baseada no artigo de Rangel “A
história da dualidade brasileira”, publicado na Revista de Economia Política (vol 1, n. 4, out.-dez./1981). 10 Rangel (1962) apud Bielschowsky (1996, p. 215). CICLOS E DUALIDADE A mudança do lado interno do polo, se-
gundo Rangel (1981), só poderia acontecer se os escravos fossem incapazes de
garantir a autossuficiência, a qual estava ligada tanto ao número de cativos quan-
to à disponibilidade de terras. Ambas as condições passaram a existir com a
proibição do tráfico (1850) e com a lei que garantia que todas as terras devolutas
passassem a pertencer aos fazendeiros. Abria-se, portanto, a possibilidade de mu-
dança de dualidade. Revista de Economia Política 34 (4), 2014 • pp. 544-564 549 A segunda dualidade brasileira tem lugar na fase recessiva do 2º Kondratieff,
datada do início dos anos 1870. Nela, o escravo passa a exercer um papel análogo
ao do “servo da gleba” do feudalismo europeu. Na medida em que o servo está
interessado no resultado do seu trabalho, passa a ter uma produtividade muito
maior do que a do escravo. A fazenda tende a tornar-se internamente feudal e ex-
ternamente uma empresa comercial. Paralelamente, o senhor de escravos se con-
vertia em “senhor feudal” no lado interno do polo interno e “vassalo” do comer-
ciante no polo externo. Havia agora, portanto, um elemento comum nos dois polos
da dualidade: o capitalismo mercantil. A classe dos comerciantes nascida nos primórdios do século XIX
em uni A segunda dualidade brasileira tem lugar na fase recessiva do 2º Kondratieff,
datada do início dos anos 1870. Nela, o escravo passa a exercer um papel análogo
ao do “servo da gleba” do feudalismo europeu. Na medida em que o servo está
interessado no resultado do seu trabalho, passa a ter uma produtividade muito
maior do que a do escravo. A fazenda tende a tornar-se internamente feudal e ex-
ternamente uma empresa comercial. Paralelamente, o senhor de escravos se con-
vertia em “senhor feudal” no lado interno do polo interno e “vassalo” do comer-
ciante no polo externo. Havia agora, portanto, um elemento comum nos dois polos
da dualidade: o capitalismo mercantil. A classe dos comerciantes, nascida nos primórdios do século XIX — em uni-
dade com a burguesia industrial que fazia parte do lado externo do polo externo
— encontrava-se agora suficientemente madura para influenciar politicamente os
negócios do Estado. Havendo a segunda dualidade nascida na fase recessiva do
ciclo longo, esta deveria promover alguma forma de substituição de importações. Caberia ao capital mercantil promovê-la, basicamente incentivando a industriali-
zação interna através de processos artesanais e manufatureiros. 12 “A produção brasileira, que havia aumentado de 3,7 milhões de sacas (de 60 kg) em 1880-81 para
5,5 em 1890-91, alcançaria em 1901-02 16,3 milhões” (Furtado, 1985, p. 177). 13 “A produção de café, em razão dos estímulos oficiais recebidos cresceu fortemente [...] Entre 1925 e
1929 tal crescimento foi quase cem por cento [...] Enquanto aumenta dessa forma a produção, mantêm-
se praticamente estabilizadas as exportações. Em 1927-29 as exportações apenas conseguiam absorver
duas terças partes da quantidade produzida”. Ibidem, p. 181. CICLOS E DUALIDADE Costuma-se datar
a última década desse século como a do nascimento da indústria brasileira. A pro-
clamação da República (1889) representou a homologação das mudanças ocorridas
na segunda dualidade. Ao iniciar-se a fase “a” do 3º ciclo longo, na segunda metade do século XIX e
que se prolonga até a I Guerra Mundial (1914-1918), a produção de café no Brasil
cresceria em progressão geométrica12. Após o final da guerra, ocorre a passagem
para a fase “b” do 3º Kondratieff, que seria marcada por dois acontecimentos de
grande repercussão mundial: A Grande Depressão (1929) e a II Guerra Mundial
(1939-1945). A terceira dualidade brasileira, portanto, seria inaugurada justamen-
te numa ocasião em que a produção de café se encontrava no auge e que a deman- Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 550 da externa tendia a se retrair13. A principal mudança foi passagem do capital in-
dustrial para o lado interno do polo externo da dualidade em substituição ao
capital mercantil. Sua exclusão do lado externo do polo externo, por sua vez, pos-
sibilitaria que outra formação (superior, porque o capitalismo hegemônico está
sempre mais avançado) ocupasse o seu espaço, neste caso a do capital financeiro14. b
d
d
l d d
b
d
l da externa tendia a se retrair13. A principal mudança foi passagem do capital in-
dustrial para o lado interno do polo externo da dualidade em substituição ao
capital mercantil. Sua exclusão do lado externo do polo externo, por sua vez, pos-
sibilitaria que outra formação (superior, porque o capitalismo hegemônico está
sempre mais avançado) ocupasse o seu espaço, neste caso a do capital financeiro14. da externa tendia a se retrair13. A principal mudança foi passagem do capital in-
dustrial para o lado interno do polo externo da dualidade em substituição ao
capital mercantil. Sua exclusão do lado externo do polo externo, por sua vez, pos-
sibilitaria que outra formação (superior, porque o capitalismo hegemônico está
sempre mais avançado) ocupasse o seu espaço, neste caso a do capital financeiro14. Embora como sócio menor da terceira dualidade, a burguesia industrial sur-
gida como dissidência da classe dos comerciantes teria papel fundamental para
alavancar o processo de substituição de importações. 14 O capital financeiro a que Rangel se refere na terceira dualidade é no sentido atribuído por Hilferding,
ou seja, surgido da união do capital industrial com o capital bancário, com a dominância do primeiro
sobre o segundo. Não é como ocorre hoje em dia, onde os papéis se encontram invertidos. 15 Sobre o processo de substituição de importações, ver: Conceição Tavares (1974), Furtado (1985) e
Cardoso de Mello (1982), entre outros. 13 “A produção de café, em razão dos estímulos oficiais recebidos cresceu fortemente [...] Entre 1925 e
1929 tal crescimento foi quase cem por cento [...] Enquanto aumenta dessa forma a produção, mantêm-
se praticamente estabilizadas as exportações. Em 1927-29 as exportações apenas conseguiam absorver
duas terças partes da quantidade produzida”. Ibidem, p. 181.
14 O capital financeiro a que Rangel se refere na terceira dualidade é no sentido atribuído por Hilferding,
ou seja, surgido da união do capital industrial com o capital bancário, com a dominância do primeiro
sobre o segundo. Não é como ocorre hoje em dia, onde os papéis se encontram invertidos.
15 Sobre o processo de substituição de importações, ver: Conceição Tavares (1974), Furtado (1985) e
Cardoso de Mello (1982), entre outros. CICLOS E DUALIDADE Como estudado por vários
autores15, o esgotamento das reservas cambiais e a escassez de divisas criaram,
automaticamente, uma reserva de mercado para a produção nacional substitutiva
de importações. Segundo Conceição Tavares (1974), quando se inicia a Grande
Depressão, o Brasil dispunha de um mercado interno relativamente amplo e uma
estrutura industrial que, embora incipiente, possuía uma relativa diversificação. Isso possibilitou a gradativa substituição de importações de bens não duráveis ou
simples para bens duráveis ou complexos. Mais adiante, no pós-guerra (ao iniciar-
-se a fase “a” do 4º Kondratieff) esse esforço de substituição de importações, com
o apoio decisivo de recursos do Estado, alcançaria os bens de capital. O processo de substituição de importações não só não é interrompido na ter-
ceira dualidade como, a partir dela, a economia brasileira passaria a produzir o seu 551 Revista de Economia Política 34 (4), 2014 • pp. 544-564 próprio ciclo endógeno16. Não se tratava do ciclo longo, causado pela gestação e
propagação de novas tecnologias, que é, por sua própria natureza, prerrogativa dos
países industrializados integrados ao centro dinâmico. Tais ciclos seriam inerentes
à construção do capitalismo industrial e seriam semelhantes aos chamados ciclos
médios ou ciclos de Juglar, com duração aproximada de uma década. Após uma
fase ascendente, a economia entra em crise, a qual induz algumas mudanças insti-
tucionais, em especial no mercado financeiro, estimulando novos investimentos em
segmentos econômicos não modernizados. Os efeitos dessa “onda de investimentos”
tendem a se propagar por todo o sistema econômico, o qual entra novamente numa
fase ascendente. Como a economia brasileira mergulha numa nova fase descendente? Isso se dá
através de um processo que Rangel chamou de “dialética da capacidade ociosa” 17. Após certo tempo, as atividades que receberam investimentos geram capacidade ocio-
sa enquanto outras atividades persistem como pontos de estrangulamento. O “choque
dos contrários” entre esses dois polos do sistema econômico — um carregado de
ociosidade e outro de antiociosidade — engendrará tensões sociopolíticas que, por
sua vez, promoverão novas mudanças institucionais que irão desencadear novos in-
vestimentos, os quais deslocarão a economia novamente para uma fase ascendente. A
teoria do ciclo endógeno de Rangel procurou demonstrar que, ao contrário do que
muitos supunham, o modelo de substituição de importações não tinha se esgotado. 17 “[...] uma vez cumprido o programa de substituição de importações [...] descobríamos que a
insuficiência da capacidade para importar ressurgia sob a forma de demanda insatisfeita de um grupo
novo de produtos [grifo do autor]. Este é o núcleo do problema que proponho que se estude sob a
rubrica de dialética da capacidade ociosa, manifestada pela circunstância fundamental de que, no
próprio ato de implantar-se, engendrava o seu contrário, tendendo, portanto, a perpetuar o esforço de
desenvolvimento” (Rangel, 1980, p. 132-3). 16 “Outra singularidade da terceira dualidade está no fato de que, embora havendo começado nas
condições da fase recessiva do ciclo longo, a industrialização substitutiva de importações [...] não se
interrompeu com a passagem ascendente do 4º Kondratieff. O dinamismo do processo de industrialização,
engendrando demandas de importações sempre novas, fez com que o impulso se mantivesse” (Rangel,
1981, p. 30). Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 CICLOS E DUALIDADE semissalariato
lado interno:
lado externo:
lado externo:
rural
semicapitalismo
lado interno:
capital industrial
capital
financeiro
polo interno
polo externo
4ª dualidade
16 “Outra singularidade da terceira dualidade está no fato de que, embora havendo começado nas
condições da fase recessiva do ciclo longo, a industrialização substitutiva de importações [...] não se
interrompeu com a passagem ascendente do 4º Kondratieff. O dinamismo do processo de industrialização,
engendrando demandas de importações sempre novas, fez com que o impulso se mantivesse” (Rangel,
1981, p. 30). 17 “[...] uma vez cumprido o programa de substituição de importações [...] descobríamos que a
insuficiência da capacidade para importar ressurgia sob a forma de demanda insatisfeita de um grupo semissalariato
lado interno:
lado externo:
lado externo:
rural
semicapitalismo
lado interno:
capital industrial
capital
financeiro
polo interno
polo externo
4ª dualidade 16 “Outra singularidade da terceira dualidade está no fato de que, embora havendo começado nas
condições da fase recessiva do ciclo longo, a industrialização substitutiva de importações [...] não se
interrompeu com a passagem ascendente do 4º Kondratieff. O dinamismo do processo de industrialização,
engendrando demandas de importações sempre novas, fez com que o impulso se mantivesse” (Rangel,
1981, p. 30). 17 “[...] uma vez cumprido o programa de substituição de importações [...] descobríamos que a
insuficiência da capacidade para importar ressurgia sob a forma de demanda insatisfeita de um grupo
novo de produtos [grifo do autor]. Este é o núcleo do problema que proponho que se estude sob a
rubrica de dialética da capacidade ociosa, manifestada pela circunstância fundamental de que, no
próprio ato de implantar-se, engendrava o seu contrário, tendendo, portanto, a perpetuar o esforço de
desenvolvimento” (Rangel, 1980, p. 132-3). Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 552 A quarta dualidade brasileira, que seria também a última, ainda não havia se
completado à época em que Rangel estudava o fenômeno. O ano de 1973 (1º cho-
que do petróleo), segundo ele, marcaria a entrada dos países do centro capitalista
hegemônico na fase “b” do 4º ciclo de Kondratieff. No Brasil, a fase descendente
do ciclo longo coincidiria com a fase “b” do seu ciclo médio endógeno, pondo
termo no chamado “Milagre Econômico” (1968-1973). 18 A expressão “marcha forçada” como característica do período 1974-1979 se deve a Castro (1985).
19 “[...] o mesmo processo recessivo, que se faz acompanhar de exacerbação do processo inflacionário,
também desencadeia um processo de ‘valorização’ da terra, no sentido de elevação dos preços desse
fator, em óbvio descompasso com a renda territorial (grifo do autor). Por outras palavras, os títulos
fundiários comportam-se em contracorrente com os mobiliários. A mesma recessão que deprime os
valores mobiliários impele para cima os títulos fundiários” (Rangel, 1982a, p. 67). Revista de Economia Política 34 (4), 2014 • pp. 544-564 20 “Há tempos que o problema da terra, no Brasil, tomou a forma de um problema financeiro [grifo do
autor]. Assim, a configuração gradativa do novel capitalismo financeiro trará em seu bojo uma reforma
agrária, nada menos. Noutros termos, a reforma agrária que não precedeu a industrialização, como
julgávamos nós, os revolucionários dos anos [19]30, terá de vir agora, como coroamento da mesma
industrialização” (Rangel, 1985, p. 21). CICLOS E DUALIDADE A partir daí o esforço do
II PND (governo Geisel) de manter o crescimento “em marcha forçada”18 de forma
a completar a substituição de importações no Departamento I da economia (pro-
dução de bens de capital) e insumos importados (principalmente petróleo) desem-
bocaria numa grave crise do balanço de pagamentos, a partir do início dos anos
1980 do século passado. Diante da incapacidade de continuar financiando o crescimento da economia
com capital externo, esse financiamento deveria ser feito com poupança interna. Nessa fase, diante do avanço de sua industrialização, o Brasil estaria deixando de
produzir muitas coisas importadas simplesmente pela falta de um setor de interme-
diação financeira capaz de financiar a acumulação de capital. O “Milagre Econô-
mico” havia levado a algumas atividades econômicas a crescerem além dos limites
impostos pelo mercado, gerando, portanto, sobreacumulação de capital. Era pre-
ciso fazer com que essas atividades detentoras de poupança, mas impossibilitadas
de investir em suas próprias instalações, fossem estimuladas a transferir recursos
para outras atividades carregadas de antiociosidade, que Rangel identificou nos
serviços públicos. Sua sugestão, na época, foi a criação de uma nova lei de conces-
são de serviços públicos, que permitisse ao setor privado encarregar-se, de forma
crescente, pelos investimentos públicos de infraestrutura. A burguesia industrial, finalmente, se tornaria o sócio maior na quarta duali-
dade, representando o polo externo, e tendo como sócio menor a nova burguesia
rural, representando o polo interno. Para viabilizar a mudança no polo interno da
dualidade, seria preciso eliminar os últimos resquícios do latifúndio feudal — con-
dição necessária para criar empregos no meio rural. Nesse sentido, Rangel advoga
a diminuição do preço da terra. Na sua visão, o preço da terra no Brasil não de-
corre do seu uso enquanto tal, mas da sua utilização como especulação financeira. A estagnação econômica seguida de inflação (na fase descendente do ciclo endóge-
no) desencadeia um processo de valorização dos ativos reais, como a terra, que
nada tem a ver com o seu valor territorial. Nesse caso, os títulos fundiários con-
correm com os títulos mobiliários19. Tendo discordado da esquerda, nos anos 1950, do papel prioritário da reforma 553 Revista de Economia Política 34 (4), 2014 • pp. 544-564 agrária para desenvolver o Brasil — dado que o processo de substituição de impor-
tações criava uma reserva de mercado para a indústria — a partir da quarta dua-
lidade, a reforma agrária tornava-se imperativa20. 21 Já no final do século XVI, os escritos mercantilistas faziam relação da inflação com as reservas de
ouro de um país e seu balanço de pagamentos. Entre os chamados filósofos políticos de meados do
século XVIII, críticos dos mercantilistas e precursores dos economistas clássicos, a correlação entre
moeda e preços era ainda mais visível (em especial, em David Hume). 22 Comissão Econômica para a América Latina e o Caribe, órgão ligado às Nações Unidas, sediado em
Santiago do Chile. Fundada em 1948, tendo à frente o economista argentino Raúl Prebisch, seu objetivo
principal era desenvolver um marco teórico alternativo (Desenvolvimentismo) para a América Latina,
até então dominado por teorias construídas com base no cenário das nações que detinham a hegemonia
econômica. INFLAÇÃO: NEM MONETARISTA, NEM ESTRUTURALISTA Embora a teoria quantitativa da moeda tenha origens remotas21, os seus prin-
cípios básicos foram elaborados, no final do século XIX e início do século XX, por
Wicksel (na Suécia), que relacionou a inflação com a taxa de juros, Marshall (na
Inglaterra), que explicou a inflação pelo princípio da oferta e procura, e Fisher (nos
Estados Unidos), cuja contribuição passou a ser um “divisor de águas” para todos
os demais autores monetaristas que se seguiram, sendo Milton Friedman o mais
importante deles. A equação de trocas, de Fisher, é dada por MV = PT, onde M representa a
quantidade de moeda, V é a velocidade de circulação da moeda, P é o nível geral
de preços e T é o total de transações realizadas na economia em certo período
(produto). Pela hipótese clássica de pleno emprego, o produto não se altera, de
modo que T pode ser considerado estável no curto prazo. Levando em conta que
as condições institucionais que determinam o número de vezes que a moeda troca
de mãos mudam muito lentamente ao longo do tempo, então V também pode ser
considerado como uma constante no curto prazo. Nessas condições, é a quantida-
de de moeda M — que representa a demanda — é responsável pela alteração no
nível geral de preços P. Já a análise estruturalista da inflação surgiu dentro do referencial teórico de-
senvolvido pela CEPAL22, cujas teses principais foram resumidas por Sunkel (1962)
através de um esquema geral para a análise da inflação. Esse esquema, idealizado
com base na economia chilena, se propunha a identificar as pressões inflacionárias
e os seus mecanismos de propagação. As pressões inflacionárias básicas refletiam 21 Já no final do século XVI, os escritos mercantilistas faziam relação da inflação com as reservas de
ouro de um país e seu balanço de pagamentos. Entre os chamados filósofos políticos de meados do
século XVIII, críticos dos mercantilistas e precursores dos economistas clássicos, a correlação entre
moeda e preços era ainda mais visível (em especial, em David Hume). 22 Comissão Econômica para a América Latina e o Caribe, órgão ligado às Nações Unidas, sediado em
Santiago do Chile. Fundada em 1948, tendo à frente o economista argentino Raúl Prebisch, seu objetivo
principal era desenvolver um marco teórico alternativo (Desenvolvimentismo) para a América Latina,
até então dominado por teorias construídas com base no cenário das nações que detinham a hegemonia
econômica. 23 “[...] Cumpre notar que essa ortodoxia verdadeira não é privilégio nem de esquerda, nem da direita,
sendo, ao contrário, um fundo comum da ciência econômica, pois as equações de Fisher já estavam
perfeitamente formuladas nos parágrafos de Marx, e não há, no mundo moderno, nenhuma ortodoxia
mais verdadeiramente ortodoxa do que a dos economistas soviéticos” (Rangel, 1962, p. 267). Dizer isso
hoje é fácil, mas não naquela época em que o conflito ideológico entre esquerda e direita era muito
acirrado. INFLAÇÃO: NEM MONETARISTA, NEM ESTRUTURALISTA Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 554 a incapacidade dos setores produtivos de atender ao crescimento da demanda, em
especial a agricultura devido à inelasticidade da oferta de alimentos. A palavra “ilusão”, usada por Rangel (1978) para caracterizar tanto a visão
monetarista quanto a estruturalista, dá bem uma ideia do quanto não acreditava
no poder de explicação dessas escolas para explicar a inflação brasileira nos anos
1960. Um ano antes de publicar o seu clássico A inflação brasileira, escreveu um
artigo com esse mesmo título em que ironizava a ortodoxia, seja de direita, seja de
esquerda23. Nossos ortodoxos monetaristas (de direita), dizia ele, não percebem que
a moeda é passiva, ou seja, é o aumento de preços que leva as autoridades mone-
tárias a aumentarem a quantidade de moeda e não o contrário. Nossos ortodoxos
estruturalistas (de esquerda), por outro lado, “tomaram a nuvem por Juno” ao não
perceberem que é a estrutura concentrada de intermediação que, ao não repassar
os aumentos de preços obtidos ao nível do consumidor final para o produtor, de-
sestimula o aumento da produção por parte deste último e não uma suposta ine-
lasticidade da oferta agrícola (Rangel, 1978, p. 29). Ao convergirem para a tese das “inelasticidades da oferta” — os monetaristas
com base na hipótese do pleno emprego clássica; os estruturalistas identificando
“ponto de estrangulamento” na agricultura — ambas as correntes concluíram, equi-
vocadamente, tratar-se de inflação causada por excesso de demanda quando, ao
contrário, existia insuficiência de demanda24. O que os representantes dessas cor-
rentes não perceberam é a existência de capacidade ociosa e, mais ainda, “que a
inflação cresce com a capacidade ociosa, quando devera regredir” (Rangel, 1978, p. 15). O que ocorre é que a inflação se expande a “demanda nominal”, mas não alte-
ra a “demanda a preços constantes, a qual consequentemente, não é nem maior, nem
menor, do que antes” (Rangel, 1962, p. 269). Se a demanda real se alterasse, condu-
ziria a uma expansão da renda e não a uma elevação dos preços, conclui Rangel. Por que, para Ignácio Rangel, o monetarismo não passa de uma “ilusão”? Para chegar lá, retornemos à equação de trocas de Fisher. Invertendo o significado
da equação, suponha que a variação no volume de moeda seja determinada pela
elevação dos preços. 24 “Porque a verdade é que o Brasil é um país de baixíssima propensão a consumir — e nem se pode
conceber que seja de outro modo, dado o atual esquema de (má) distribuição de sua renda” (Rangel,
1978, p. 35). 26 A primeira edição de A inflação brasileira é de 1963, quando a obra do economista polonês era pouco
conhecida no Brasil. Por esse motivo, no prefácio da 3ª edição (1978), Luiz Carlos Bresser-Pereira não
acredita que Ignácio Rangel estivesse familiarizado com a teoria kaleckiana. O próprio Rangel, no
posfácio que escreveu para a edição de 1978, mostrou estar em dúvida a esse respeito. “[...] tanto podia
dizer que sim, como que não” (Rangel, 1978, p. 129). INFLAÇÃO: NEM MONETARISTA, NEM ESTRUTURALISTA Nessa hipótese, a igualdade da equação de Fisher MV = PT se
converte na desigualdade MV < P1T. Essa hipótese não foi considerada pelos mo-
netaristas porque, como se trata de uma “equação de trocas”, para reestabelecer a
igualdade entre o lado direito e esquerdo da equação, uma parte da produção não
entraria no mercado, tal que MV= P1 (T – t), onde t é a parte do produto físico re- Revista de Economia Política 34 (4), 2014 • pp. 544-564 555 tirada do mercado. Em concorrência pura, como supõem os monetaristas, isso não
poderia acontecer. Mas numa economia de características monopolistas como a
brasileira, a hipótese de retenção voluntária da oferta ou de capacidade ociosa
planejada passa a ser perfeitamente plausível. Essa retenção de parte da produção pelas empresas, caso persista por muito
tempo, resultaria em queda da renda real pelo aumento do nível dos estoques. Se
isso não ocorre é porque a elevação dos preços (de P para P1) para provoca altera-
ção no primeiro membro do lado esquerdo da equação de trocas (de M para M1),
que poderia ser reescrita como M1V= P1 T. Em outras palavras, a retirada de parte
do produto do mercado (t), alterando o total de bens e serviços oferecidos ao mer-
cado para T – t, sustenta o acréscimo no preço P1. A emissão de moeda passa a ser,
portanto, consequência ao invés de causa da inflação. O aumento do meio circu-
lante pelo governo, de M para M1 destina-se a suprir as necessidades de caixa das
empresas devido ao alto custo de manutenção de estoques, reestabelecendo o equi-
líbrio da equação de trocas a um nível maior de inflação. O papel do governo
nesse processo, portanto, é passivo25. O problema fundamental, para Ignácio Rangel, é descobrir por que algumas
empresas privadas que dominam o mercado, em certos períodos, decidem elevar os
seus preços — e mais — por que preferem até mesmo não levar uma parte do pro-
duto ao mercado ao invés de baixarem os preços? Ora, à medida que nos afastamos
do regime de concorrência pura e adotamos a tese mais realista da concentração
de mercado, a manutenção da margem de lucro (regra do mark-up), em períodos
de escassez da demanda, passa a ser uma explicação convincente. 25 “Tudo se passa, portanto, como se a sociedade civil, movida por maquiavélica malícia, preparasse
uma armadilha para o Estado. Este será premiado se desempenhar o seu papel na tragicomédia da
inflação; e será punido se recusar a fazê-lo” (Rangel, 1978, p. 26). 27 Não é outro o pensamento de Rangel, como pode ser comprovado pela leitura do prefácio do livro
Recursos ociosos e política econômica (1980, p. xi): “[...] o progresso não depende, necessariamente, da
expansão das forças produtivas, dado que, normalmente (grifo do autor), a sociedade subutiliza as
forças de que dispõe. Assim, quando o marxismo me ensinou que toda crise é crise de superprodução,
disse-me uma antiga novidade”. INFLAÇÃO: NEM MONETARISTA, NEM ESTRUTURALISTA Nesse ponto, a teoria de formação de preços de Ignácio Rangel em muito se
assemelha a de Michal Kalecki, embora não seja certo que ele conhecesse a obra
do economista polonês26 à época (início dos anos 1960). Mesmo não conhecendo
Kalecki, a afinidade provavelmente se deu porque ambos tiveram o mesmo ponto
de partida (Marx). Para Kalecki, a característica normal da economia é operar com
capacidade ociosa devido ao grau de monopolização das economias capitalistas27. O preço do produto (p) é determinado por uma margem (mark-up) sobre os custos
diretos (u), calculado pela equação p = m u + n p*, sendo m e n coeficientes posi- 27 Não é outro o pensamento de Rangel, como pode ser comprovado pela leitura do prefácio do livro
Recursos ociosos e política econômica (1980, p. xi): “[...] o progresso não depende, necessariamente, da
expansão das forças produtivas, dado que, normalmente (grifo do autor), a sociedade subutiliza as
forças de que dispõe. Assim, quando o marxismo me ensinou que toda crise é crise de superprodução,
disse-me uma antiga novidade”. Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 556 tivos que representam a política de fixação de preços da firma e p* o preço dos
demais concorrentes naquele mercado. O poder de mercado da empresa reflete o
seu grau de monopólio, dado pela razão p
u . Quanto menor o poder de mercado da
empresa, mais próximo de p* deverá situar-se seu preço e vice-versa28. Conquanto a escola estruturalista tenha entendido, corretamente, a inflação
como um fenômeno endógeno, também, para Ignácio Rangel, produziu outra “ilu-
são”. Ao associar o aumento dos preços com os “gargalos” da estrutura da econo-
mia, notadamente a inelasticidade da oferta agrícola, os estruturalistas inverteram
a ordem natural das coisas: a inelasticidade está na demanda e não na oferta. É o
intermediário oligopsonista-oligopolista que impede que a elevação dos preços
pagos pelo consumidor seja transferida para o produtor. Como a elevação de preços
deprime o salário real do trabalhador — dado o peso da alimentação no orçamen-
to das classes de renda mais baixas — o que existe é uma crônica inelasticidade da
demanda e não da oferta. A parte mais surpreendente do raciocínio de Rangel ainda não foi desvendada. A redução da demanda não ocorre nos chamados bens-salário e sim em outros bens,
cuja procura é mais elástica do que os alimentos, tais como vestuário, calçados, etc. 28 Ver Kalecki (1983, p. 8). INFLAÇÃO: NEM MONETARISTA, NEM ESTRUTURALISTA Em outras palavras, o aumento da parcela da renda do trabalhador comprometida
com a alimentação provoca um efeito substituição na estrutura de consumo popu-
lar, aumentando o peso dos alimentos em detrimento de outras mercadorias e
serviços menos essenciais. O aumento dos estoques, portanto, não se verifica no
segmento de empresas que aumentaram os preços dos seus produtos e sim nas
empresas cuja elasticidade preço da demanda é maior. Os preços não caem, como
supõem os monetaristas, porque a elevação inicial de preços e a retenção de esto-
ques têm lugar em setores diferentes do mercado e são estas últimas empresas que
recorrem ao sistema bancário à procura de crédito para manter os estoques que
forçam o governo a emitir, sancionando a inflação. Desfeitas as “ilusões” monetarista e estruturalista, Ignácio Rangel passa a ex-
por o seu próprio pensamento a respeito das causas do processo inflacionário
brasileiro da primeira metade dos anos 1960. Suas referências teóricas são Keynes
e Marx. Do primeiro, foi buscar o conceito de “propensão marginal a consumir”
para explicar a proporção da renda que é gasta com bens de consumo. Do segundo,
se apropriou do conceito de “taxa de exploração” — proporção do excedente ou
mais-valia realizada sobre os salários pagos pelo trabalho — para entender o pro-
cesso de concentração de renda no país. Juntando os dois conceitos, Rangel con-
cluiu que pelo fato da taxa de exploração ser alta é que a propensão marginal a
consumir é baixa. Um passo mais adiante e Rangel consegue explicar a recorrência de crises
econômicas ou, como costumava chamar, as fases descendentes dos ciclos econô-
micos. Na medida em que é apropriada uma fatia maior da mais-valia (ou que
aumenta a taxa de exploração), as condições para aumento do investimento tendem Revista de Economia Política 34 (4), 2014 • pp. 544-564 557 a se deprimir por causa da insuficiência de demanda efetiva. Esta queda, por sua
vez, resulta do aumento de produtividade não repassada para os salários. O dilema
da economia brasileira reside, para Rangel, entre escolher entre crescimento cada
vez mais acelerado — de forma a ocupar a grande carga de capacidade ociosa
acumulada em seu parque industrial — e a crise econômica. Finalmente, Rangel conclui que a inflação é uma espécie de mecanismo de
sustentação do nível de crescimento da economia. INFLAÇÃO: NEM MONETARISTA, NEM ESTRUTURALISTA Para que a economia cresça é
preciso manter um nível mínimo de demanda, o que só é possível devido à existên-
cia da inflação. A elevação de preços tem como contrapartida a desvalorização da
moeda e, com isso, provoca uma “corrida aos bens materiais”, promovendo uma
elevação forçada dos investimentos pelas empresas e das compras de bens duráveis
pelas famílias de renda média e alta. A inflação, portanto, está relacionada com
aquilo que mais estratégico existe para uma economia, ou seja, a sua própria taxa
de formação de capital29. 29 “[...] Vista sob esse ângulo, a inflação se relaciona não com movimentos superficiais da economia,
mas com o que há de mais estratégico nela, isto é, com a taxa de capitalização ou de formação de capital”
(Rangel, 1978, p. 32). 30 No seu período de auge (1970-1973), o PIB brasileiro cresceu, em média, acima de 10% ao ano e a
produção industrial cresceu, também em média, 13% ao ano (Rangel, 1985a). “MILAGRE” E “ANTIMILAGRE” O “Milagre Econômico” (1968-1973) ficou marcado como um período de alto
crescimento da economia brasileira, sobretudo entre 1970 e 197330. Para Rangel
(1985a), entretanto, esse período nada teve de “milagroso”. Foi simplesmente uma
decorrência de uma conjuntura extremamente favorável, marcada pela coincidência
da fase “a” do seu ciclo endógeno com a etapa final da mesma fase do 4º Ciclo
Longo (ou de Kontradieff). Não sendo apenas uma economia reflexa, a partir do
início deste Ciclo Longo, haveria fatores internos que seriam coadjuvantes do pro-
cesso de recuperação da economia brasileira. Dentre estes, Rangel destaca a implan-
tação da correção monetária (curiosamente, a “indexação” seria eleita como causa
da hiperinflação dos anos 1980). Porém, assim como a Instrução 70, de Eugênio
Gudin, teve muito a ver com a fase de crescimento do governo JK; a correção mo-
netária, de Octávio Bulhões, foi responsável pelo “Milagre” de Delfim Netto. A
partir de 1973, quando a economia capitalista entraria na fase “b” do Ciclo Longo,
o Brasil ainda permaneceria na fase ascendente do seu ciclo breve até o final da
década, situação que mudaria drasticamente a partir do início dos anos 1980. Consciente de que o Brasil se encontrava na fase descendente do seu ciclo
médio endógeno que, pela primeira vez, coincidia com a fase “b” do ciclo longo
exógeno, o interesse de Rangel volta-se para a conjuntura econômica, particular- Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 558 mente na busca de saída para a estagflação que assolou o país na década de 198031. Acertadamente, Rangel previu que a recuperação de 1984 não representava uma
passagem para a fase ascendente do ciclo endógeno e sim um efeito induzido de
uma temporária recuperação mundial relacionada com o déficit do Tesouro norte-
-americano. O financiamento do déficit, a juros elevados, se constituía num verda-
deiro “aspirador” de recursos do resto do mundo, praticamente inviabilizando aos
países dependentes altamente endividados — como o Brasil — de financiarem os
seus déficits de Balanço de Pagamentos. Nesse sentido, a economia brasileira, que
contava com considerável reserva de capacidade ociosa herdada do ciclo anterior,
viu-se forçada a buscar divisas através de superávits na sua balança comercial para
contrabalançar a falta de financiamento externo. Portanto, 1984 foi apenas um
suspiro de recuperação que não duraria muito. A década de 1980 marcou também o início do processo de aumento dos preços
que culminou com hiperinflação. 31 Ver, especialmente, o artigo “Recessão, inflação e dívida interna”, publicado no segundo semestre de
1985, na Revista de Economia Política (vol. 5, n. 3, jul.-set./1985).
32 “Trata-se, em suma, de precisar a síndrome da recessão. Os sintomas isolados que compõem essa
síndrome já se encontram razoavelmente cobertos, mas cada especialista tende a privilegiar o aspecto
que, per faz et nefaz, obteve a sua atenção, perdendo de vista a floresta, por causa das árvores” (Rangel,
1982a, p. 65). “MILAGRE” E “ANTIMILAGRE” Rangel discordava da tendência dominante entre
os economistas de que a inflação era causada por “excesso de demanda” e via a
inflação com um componente da “síndrome da recessão”32. Somente se poderia
falar em “aquecimento” da demanda na fase ascendente do ciclo, mas não na fase
descendente, quando a demanda é mínima. Ora, na recessão, a tendência é haver
subutilização da capacidade instalada, de modo que a redução da oferta é, até
certo ponto, planejada. Contudo, o que leva o empresário a operar com capacida-
de ociosa e gerar aumento dos preços? Ocorre que, numa economia oligopolizada
como a brasileira, quando a capacidade instalada ultrapassa a demanda, as empre-
sas, defensivamente, reduzem a oferta e aumentam os preços para manter a sua
margem de lucro. Essa é a explicação. Quando Rangel diz que a inflação faz parte da “síndrome da recessão” ele quer
dizer que não há como baixar a inflação sem atacar a causa da recessão. Nesse
ponto, entra em cena a “dialética da capacidade ociosa”. Na recessão (ou fase “b”
do ciclo endógeno), existe um “polo de ociosidade”, composto de atividades que
estiveram se modernizando prioritariamente na fase ascendente do ciclo, e um
“polo de antiociosidade”, do qual fazem parte atividades atrasadas tecnologicamen-
te e cujo desenvolvimento passa a ser exigido pela sistema para sair da recessão. O
nó a ser desatado consiste em criar condições para investimento de recursos das
empresas daquelas atividades carregadas de ociosidade para os setores retardatários. Que setores seriam esses? Os grandes serviços de utilidade pública, como transpor-
tes pesados de carga, transportes de massa de passageiros (metrôs), energia, serviços
urbanos, etc. Admitindo que, na época, o setor de intermediação financeira ainda Revista de Economia Política 34 (4), 2014 • pp. 544-564 559 não estava preparado para transferir poupança de um setor para outro, a solução
do problema teria de passar pela intervenção do Estado. p
p
p
O problema, segundo Rangel (1985), admite duas soluções: a) a poupança
privada seria transferida (via dívida pública) para o Estado que se encarregaria de
investir nas atividades retardatárias; b) ao Estado apenas caberia criar condições
propícias ao investimento do setor privado nos serviços públicos, mediante mudan-
ças no arcabouço legal. Desde logo, Rangel descarta a primeira alternativa, por se
colocar no longo prazo, enquanto o problema exige solução de curto prazo. 34 “A presente crise deverá desembocar numa solução desse segundo tipo. Isso implicará, numa primeira
etapa, a progressiva participação do capital privado na implantação de projetos rompedores de pontos
de estrangulamento; [...] numa segunda etapa, não é possível excluir a possibilidade, ou de privatização
pura e simples da atividade ou, dependendo das circunstâncias, de conversão dos serviços públicos de
administração direta, ou das empresas públicas, em serviços públicos concedidos a empresas privadas”
(Rangel, 1982c, p. 81). 33 Um exemplo de que a economia brasileira ainda não estava preparada para o fornecimento de tais
serviços é o da empresa estrangeira Light que, para implantar o serviço de bondes nas grandes cidades,
teve de importar desde o equipamento pesado para a geração e transporte de energia até os próprios
bondes (Rangel, 1985). 35 Ibidem, nota 30, p. 22. “MILAGRE” E “ANTIMILAGRE” Para
reforçar seu argumento, lembra que, no limiar de nossa industrialização, os atuais
serviços públicos nasceram como concessões a empresas privadas, quase sempre
estrangeiras33. Posteriormente, o Estado promoveu a nacionalização desses serviços. Agora, não deverá ser diferente. Haverá uma redistribuição de responsabilidades
entre o setor público e o setor privado, mas a “variável estratégica é a privatização”,
conclui Rangel34. A retomada do crescimento exigiria, para Rangel, uma reformulação institu-
cional do setor de intermediação financeira, de modo a possibilitar que os lucros
dos setores portadores de capacidade ociosa sejam transformados em excedente
social a ser reinvestido em projetos capazes de eliminar os “gargalos” da economia. A mudança institucional a que se refere Rangel passaria por uma generalização do
uso do “poder de aval” do Estado, mudança na qual o BNDES deveria desempenhar
“função-chave”35. Mas não basta apenas uma mudança no arcabouço legal; seria
preciso fazer com que as taxas reais de juros se transformassem de positivas em
negativas — como acontecia à época em que os serviços públicos foram estrutura-
dos no Brasil. Aqui a análise de Rangel sofre forte influência de Keynes, na medida
em que considera que o investimento só é viável se a eficácia marginal do capital
for maior que a taxa de juros de mercado. Não resta dúvida que o problema dos juros é o mais difícil de ser resolvido,
dado que o Brasil, tanto no final do século passado como no presente século, está
entre os países que apresentam as mais elevadas taxas de juros reais do planeta. Mas se há algo errado, adverte o mestre, é a existência de taxas reais de juros for-
temente positivas e não o fato de, eventualmente, serem negativas. Segundo ele, a
eficácia marginal do capital das empresas com capacidade ociosa é negativa e, 34 “A presente crise deverá desembocar numa solução desse segundo tipo. Isso implicará, numa primeira
etapa, a progressiva participação do capital privado na implantação de projetos rompedores de pontos
de estrangulamento; [...] numa segunda etapa, não é possível excluir a possibilidade, ou de privatização
pura e simples da atividade ou, dependendo das circunstâncias, de conversão dos serviços públicos de
administração direta, ou das empresas públicas, em serviços públicos concedidos a empresas privadas”
(Rangel, 1982c, p. 81). Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 560 pela lógica, é essa eficácia que deve orientar a taxa de juros. 36 “Seria fútil, ou mal-intencionado, identificar esse estado de coisas com o advento do socialismo. O
Brasil, por certo, caminha também para o socialismo, mas não pela via da mera estatização da economia,
tanto mais quanto esta, como ficou demonstrado, é uma rua de mão dupla, na qual o capitalismo
privado tem ressurgido sempre das próprias cinzas”. Ibidem, nota 31, p. 81. “MILAGRE” E “ANTIMILAGRE” Entre as causas que
contribuíram para que a taxa real de juros se tornasse positiva está o fato de que
o setor público não tem outra garantia a oferecer a não ser o aval do Tesouro. Ora,
o endividamento crescente do Estado e o risco de insolvência deste tornam esse aval
de valor discutível, o que explica a existência de taxas reais de juros positiva. Por uma simples questão de bom-senso se chega à conclusão de que nenhum
serviço público será capaz de remunerar a empresa privada com tarifas compatíveis
com as elevadas taxas de juros do mercado. Para que as taxas reais de juros sejam
menores que a eficácia marginal do capital investido nesses setores, torna-se neces-
sário que o Estado levante recursos, através de sua influência no setor financeiro
ou dos bancos públicos, e transfira-os para essas empresas com forte subsídio. Pode parecer que nada terá mudado com a passagem dos serviços públicos da
responsabilidade do Estado para a empresa privada. Porém, em troca do seu aval,
o Estado deixará de comprometer recursos fiscais futuros e ainda terá como garan-
tia a hipoteca dos bens pertencentes às empresas concessionárias. Em caso de má
prestação do serviço público, poderá o Estado executar a hipoteca do concessioná-
rio, inclusive tomando-lhe a concessão, se for o caso, e negociando-a com outro
concessionário. Em resumo, será por esta porta que nascerá o “capital financeiro brasileiro”,
isto é, o casamento entre o capital bancário remodelado e o capital industrial in-
vestido nos serviços públicos. Na aparência, nosso capital financeiro estará nascen-
do como capitalismo de Estado. Mas isso não deve ser confundido com socialismo. Rangel mantém a esperança viva de que o Brasil ainda possa caminhar para o so-
cialismo, mas não pela via da estatização da economia36. Trata-se simplesmente de
uma “rua de mão dupla”: de um lado, o setor privado, não encontrando mercado
à altura da capacidade produtiva criada na precedente fase ascendente do ciclo,
recorre ao Estado para resolver seu problema; de outro, o Estado, prestes a esgotar
a sua capacidade de endividamento, vê uma saída para a falta de recursos na trans-
ferência dos serviços públicos para a órbita privada. Em definitivo, conclui Rangel
(1982c, p. 80), “resta apenas o fato de que certas atividades, em cada momento,
estão a cargo do setor privado, enquanto outras integram o setor público. Não
necessariamente as mesmas, com o correr do tempo”. Revista de Economia Política 34 (4), 2014 • pp. 544-564 PROFETA OU VISIONÁRIO? Ao nos associarmos às comemorações do centenário do nascimento de um dos
mais originais pensadores brasileiros, que juízo podemos fazer dele? Profeta ou
visionário? Ignácio Rangel era um pouco de cada coisa. Tanto poderia ser visto Revista de Economia Política 34 (4), 2014 • pp. 544-564 561 como um indivíduo que prediz o futuro, quanto como um sonhador. A simples
comparação com o presente, que emerge da leitura das seções precedentes deste
texto, revela um Ignácio Rangel profeta. Previu, com cerca de duas décadas de
antecedência, a privatização dos serviços públicos ou a concessão de serviços pú-
blicos à empresa privada, iniciada durante o governo de Fernando Henrique Car-
doso e que chegou, em menor ou maior grau, até o governo Dilma Rousseff. Tinha
também sonhos, que alguns podem considerar como utopias, como sua inabalável
convicção de que o Brasil caminhava para o socialismo, embora talvez não no
sentido comumente associado à palavra. Sua visão de socialismo não era sinônimo
de “estatismo”, pois acreditava que os ciclos exógenos (das economia centrais) e
endógenos (determinados pela “dialética da capacidade ociosa”), mais a escassez
de recursos públicos, impunham limites a atuação do Estado no sistema de econo-
mia de mercado37. Daí haver necessidade da parceria do capital público-privado
— sempre com o controle do primeiro —, alternando as suas áreas de atuação ao
longo do ciclo econômico, seja para romper com as fases “b” dos ciclos endógenos,
seja para galgar mais rapidamente as etapas do desenvolvimento, cujo conceito
está baseado no tripé avanço tecnológico/crescimento econômico/justiça social. Mais importante do que traçar o perfil de I. Rangel, certamente multifacetado,
é resgatar a sua contribuição no terreno da economia, de sorte que possa servir de
inspiração para as novas gerações de economistas38, assim como foi para a geração
deste articulista. Formado em Direito, Rangel foi, ao longo de sua vida, um auto-
didata. Apreendeu economia por conta própria, começando por Marx, até tornar-
-se um desenvolvimentista, supostamente influenciado pela leitura de Keynes, no
único curso formal que frequentou (sobre planejamento), na CEPAL, no Chile. Ao
contrário daqueles que se dizem “economistas de esquerda”, mas que perderam a
capacidade crítica, Rangel nunca deixou de pensar dialeticamente. Do mesmo mo-
do, sem menosprezar a importância do mercado, jamais se deixou influenciar pelas
teses dos “fundamentalistas do mercado”, tão em voga nos dias de hoje39. Achar um rótulo para I. Rangel não é tarefa fácil. 37 Ignácio Rangel já pensava globalmente numa época em que a globalização ainda não fazia parte da
agenda dos economistas.
38 Na verdade, Rangel aspirava a que os jovens seguissem adiante com o seu trabalho. No prefácio do
livro Ciclo, tecnologia e crescimento (1982, p. 11), escreveu: “[...] de algum modo é mister comprometer
os quadros jovens nesta empreitada, e quem sabe se eles não encontrarão motivo de realização pessoal
aplicando-se a desmantelar os esquemas do ‘velho Rangel’”.
39 “A suposição de que o capitalismo brasileiro esteja a caminho de recompor a competição perfeita [...]
não tem coisa alguma que a corrobore” (Rangel, 1985a, p. 81). 39 “A suposição de que o capitalismo brasileiro esteja a caminho de recompor a competição perfeita [...]
não tem coisa alguma que a corrobore” (Rangel, 1985a, p. 81). PROFETA OU VISIONÁRIO? Não foi monetarista, porque
acreditava que a moeda era passiva — o governo, ao emitir moeda, apenas sancio-
nava a inflação. Tampouco era estruturalista, porque achava que o aumento dos
preços era causado pela estrutura oligopolizada de comercialização de produtos
agrícolas e não pela incapacidade da agricultura aumentar a oferta. Sequer foi key-
nesiano, porque demonstrou que, no longo prazo, a inflação tinha uma relação in-
versa (ao invés de direta) com o crescimento, coisa não admitida pelos compêndios 37 Ignácio Rangel já pensava globalmente numa época em que a globalização ainda não fazia parte da
agenda dos economistas. 39 “A suposição de que o capitalismo brasileiro esteja a caminho de recompor a competição perfeita [...]
não tem coisa alguma que a corrobore” (Rangel, 1985a, p. 81). Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 562 de macroeconomia de sua época40. Usava categorias de análise comuns ao materia-
lismo histórico, mas via o socialismo com ressalvas. Foi um heterodoxo muito antes
de essa palavra entrar na moda. Ele próprio se definia como uma economista de
esquerda, mas essa expressão não tem o significado hoje que tinha naquela época. Chega-se à conclusão que nenhum rótulo “cola” em I. Rangel. Por quê? Porque
simplesmente talvez ele sido o mais original dos nossos pensadores econômicos41. REFERÊNCIAS BIBLIOGRÁFICAS Folha de São Paulo SCHOWSKY, Ricardo (1996) Pensamento Econômico Brasileiro. Rio de Janeiro: Contraponto. BRESSER-PEREIRA, Luiz Carlos e José Márcio Rego (1993), “Um mestre da economia bra
nácio Rangel”, Revista de Economia Política, v. 13, n. 2 (50), abril-junho/1993. CARDOSO DE MELLO, J.M. (1982) O capitalismo tardio. São Paulo: Brasiliense. CASTRO, Antonio Barros de. (1985) A economia brasileira em marcha forçada. Rio de Janeiro: Paz e
Terra. CONCEIÇÃO TAVARES, Maria da (1974) Da substituição de importações ao capitalismo financeiro. Rio de Janeiro: Zahar. CONCEIÇÃO TAVARES, Maria da (1974) Da substituição de importações ao capitalismo fin
Rio de Janeiro: Zahar. olha de São Paulo (1994) “Economista Ignácio Rangel morre no Rio”, São Paulo, 5 março 1994 FURTADO, Celso (1985) Formação Econômica do Brasil, 20ª edição, São Paulo: Cia. Editora. Nacio-
nal. KALECKI, Michal (1983) Teoria da Dinâmica Econômica. Coleção “Os Economistas”. São Paulo:
Abril Cultural. PEREIRA, José Maria Dias (1984) “A participação da alimentação na inflação brasileira nos anos 70”. Porto Alegre: FEE, 1984 (Série “Teses”, n.7). PEREIRA, José Maria Dias (1993) “Os trinta anos de A Inflação Brasileira de Ignácio Rangel”, Revis-
ta de Economia Política, v. 13, n. 3 (51). julho-set./1993. RANGEL, Ignácio Mourão (1962) “A inflação brasileira”. In: Inflação e Desenvolvimento. Rio de Ja-
neiro: Vozes (extraído de “Econômica Brasileira”, abril/junho). RANGEL, Ignácio Mourão (1978) A inflação brasileira. 1963. 3ª ed. São Paulo: Brasiliens RANGEL, Ignácio Mourão (1980) Recursos ociosos e política econômica. São Paulo: Hucitec RANGEL, Ignácio Mourão (1981) “A questão financeira”, Revista de Economia Política, v. 1, n. 1. janeiro-março. RANGEL, Ignácio Mourão (1981) “A história da dualidade brasileira”, Revista de Economia Política,
v. 1, n. 4. outubro-dezembro. RANGEL, Ignácio Mourão (1982) “O Brasil na fase “b” do 4º Kondratieff”, In Ciclo, tecnologia e
crescimento. Rio de Janeiro: Civilização Brasileira. RANGEL, Ignácio Mourão (1982a.) “A síndrome da recessão brasileira” In: Ciclo, tecnologia e cresci-
mento. Rio de Janeiro: Civilização Brasileira. 40 O fenômeno da “estagflação” só passou a fazer parte dos manuais de macroeconomia a partir da
contribuição de Milton Friedman e Edmund Phelps (final dos anos 1960), que incorporaram as
expectativas de inflação na análise tradicional da curva de Phillips, demonstrando que o trade-off entre
inflação e desemprego só ocorre no curto prazo. No longo prazo, quando os agentes “adaptam as suas
expectativas”, a política macroeconômica é ineficaz. Ver Sachs-Larrain (2000). 40 O fenômeno da “estagflação” só passou a fazer parte dos manuais de macroeconomia a partir da
contribuição de Milton Friedman e Edmund Phelps (final dos anos 1960), que incorporaram as
expectativas de inflação na análise tradicional da curva de Phillips, demonstrando que o trade-off entre
inflação e desemprego só ocorre no curto prazo. No longo prazo, quando os agentes “adaptam as suas
expectativas”, a política macroeconômica é ineficaz. Ver Sachs-Larrain (2000). Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 REFERÊNCIAS BIBLIOGRÁFICAS Folha de São Paulo 41 A primeira frase de um artigo de Bresser-Pereira reforça a minha conclusão: “Ignácio Rangel é
provavelmente o mais original analista do desenvolvimento econômico brasileiro. Apenas Celso Furtado
tem uma contribuição comparável na análise da dinâmica de nossa economia” (Bresser-Pereira, 1993,
p. 98). Revista de Economia Política 34 (4), 2014 • pp. 544-564 563 RANGEL, Ignácio Mourão (1982b) “O direitismo da esquerda”. In: Ciclo, tecnologia e crescimento. Rio de Janeiro: Civilização Brasileira. RANGEL, Ignácio Mourão (1982c) “A questão financeira”. In: Ciclo, tecnologia e crescimento. Rio de
Janeiro: Civilização Brasileira. Janeiro: Civilização Brasileira. RANGEL, Ignácio Mourão (1984) “Dualidade e ciclo longo”, Revista de Economia Política, v. 4, n. 1
janeiro-março. RANGEL, Ignácio Mourão (1985) “Recessão, inflação e dívida externa”, Revista de Economia Política,
v. 5, n. 3. julho-setembro. RANGEL, Ignácio Mourão (1985a) Milagre e Anti-Milagre. Rio de Janeiro: Jorge Zahar, RANGEL, Ignácio Mourão (1990) “O quarto ciclo de Kondratiev”, Revista de Economia Política, v. 10, n. 4. outubro-dezembro. SACHS, Jeffrey e Felipe B. LARRAIN, (2000) Macroeconomia em uma economia global. São Paulo:
Pearson. SUNKEL, Osvaldo (1957) “Um esquema geral para análise da inflação”, In: Inflação e Desenvolvimen-
to. Rio de Janeiro: Vozes (extraído de “Econômica Brasileira”, julho/dez. 1957). Brazilian Journal of Political Economy 34 (4), 2014 • pp. 544-564 564
|
https://openalex.org/W4286216955
|
http://ahea.pitt.edu/ojs/index.php/ahea/article/download/468/840
|
English
| null |
Subject-Position and Béla Bartók’s Music for Strings, Percussion, and Celesta (1936) in Stanley Kubrick’s The Shining (1980)
|
Hungarian cultural studies
| 2,022
|
cc-by
| 4,568
|
Lucas, Sarah. “Subject-Position and Béla Bartók’s Music for Strings, Percussion, and Celesta (1936) in Stanley
Kubrick’s The Shining (1980).” Hungarian Cultural Studies. e-Journal of the American Hungarian Educators
Association, Volume 15 (2022): http://ahea.pitt.edu DOI: 10.5195/ahea.2022.468 Lucas, Sarah. “Subject-Position and Béla Bartók’s Music for Strings, Percussion, and Celesta (1936) in Stanley
Kubrick’s The Shining (1980).” Hungarian Cultural Studies. e-Journal of the American Hungarian Educators
Association, Volume 15 (2022): http://ahea.pitt.edu DOI: 10.5195/ahea.2022.468 Subject-Position and Béla Bartók’s Music for Strings, Percussion,
and Celesta (1936) in Stanley Kubrick’s The Shining (1980) Subject-Position and Béla Bartók’s Music for Strings, Percussion,
and Celesta (1936) in Stanley Kubrick’s The Shining (1980) Sarah Lucas Abstract: Twentieth-century art music composed by Bartók, Ligeti and Penderecki
constitutes a large portion of the soundtrack for Stanley Kubrick’s 1980 film adaptation of
Stephen King’s novel, The Shining. This music was not written for the film, and the use of
these pieces might leave listeners doubtful as to the legitimacy of a connection between
them and the scenes in the movie they were used to enhance. However, in the case of the
Bartók work excerpted in the film – Music for Strings, Percussion and Celesta (1936) – an
analysis of the subject-position of the music allows for another interpretation. Eric Clarke
identifies subject-position in music as “the way in which characteristics of the musical
material shape the general character of a listener’s response or engagement,” a definition
based on earlier explorations of subject-position in film studies. My analysis of the subject-
position of Bartók’s piece and the scenes in which excerpts of the work appear in The
Shining reveals similarities in their potential effect on an audience member. Keywords: Béla Bartók; Music for Strings, Percussion, and Celesta; The Shining; subject-
position; film music Biography: Sarah M. Lucas is a Lecturer of Music History, Music Theory and Ear Training at Texas
A&M University-Kingsville. She completed her doctoral studies in musicology at the University of Iowa. Her dissertation, Fritz Reiner and the Legacy of Béla Bartók’s Orchestral Music in the United States, is
based on archival research carried out in the U.S. and in Hungary, where she conducted research at the
Budapest Bartók Archives with the support of a Fulbright Fellowship in affiliation with the Hungarian
Institute for Musicology and the Hungarian Academy of Sciences. Her master’s work in music history at
the University of Missouri culminated in her thesis, Béla Bartók and the Pro-Musica Society: A Chronicle
of Piano Recitals in Eleven American Cities During his 1927-1928 Tour. Lucas is currently carrying out
further research on Bartók and Reiner with the support of an American Philosophical Society Franklin
Grant. She regularly presents her research at regional, national, and international conferences and has
published articles in journals such as Magyar Zene [‘Hungarian Music’] and Hungarian Cultural Studies. Most recently she has contributed to content guides for the Nineteenth-Century materials of the A-R
Music Anthology. sarah.lucas@tamuk.edu New articles in this journal are licensed under a Creative Commons Attribution 4.0 International License. This journal is published by Pitt Open Library Publishing. Subject-Position and Béla Bartók’s Music for Strings, Percussion,
and Celesta (1936) in Stanley Kubrick’s The Shining (1980) ISSN 1936-8879 (online) New articles in this journal are licensed under a Creative Commons Attribution 4.0 International License. This journal is published by Pitt Open Library Publishing. ISSN 1936-8879 (online) New articles in this journal are licensed under a Creative Commons Attribution 4.0 International License. Lucas, Sarah. “Subject-Position and Béla Bartók’s Music for Strings, Percussion, and Celesta (1936) in Stanley
Kubrick’s The Shining (1980).” Hungarian Cultural Studies. e-Journal of the American Hungarian Educators
Association, Volume 15 (2022): http://ahea.pitt.edu DOI: 10.5195/ahea.2022.468 Lucas, Sarah. “Subject-Position and Béla Bartók’s Music for Strings, Percussion, and Celesta (1936) in Stanley
Kubrick’s The Shining (1980).” Hungarian Cultural Studies. e-Journal of the American Hungarian Educators
Association, Volume 15 (2022): http://ahea.pitt.edu DOI: 10.5195/ahea.2022.468 Twentieth-century art music composed by Béla Bartók, György Ligeti and Krzysztof
Penderecki constitutes a significant portion of the soundtrack for Stanley Kubrick’s 1980 film
adaptation of Stephen King’s novel The Shining, while the remainder of the film score was
written for the movie by Wendy Carlos and Rachel Elkind. The use of preexisting music in a
film soundtrack is a topic already explored by numerous film and music scholars, but as a Bartók
specialist I was particularly intrigued by the inclusion of Bartók’s music in a horror film, as a
connection between his music and the plot of The Shining was not clear to me. Written for his
publisher, Universal Edition, Bartók’s 1937 program notes for Music for Strings, Percussion and
Celesta indicate that the piece was not conceived as a programmatic work and did not contain
intentional elements of horror (Suchoff, 2004: 32). However, unique elements of the work’s third
movement, such as its instrumentation, form, and “night music” texture, (used by Bartók as early
as 1926 to depict natural sounds of the night, here, featuring glissandos, string tremolos, Twentieth-century art music composed by Béla Bartók, György Ligeti and Krzysztof
Penderecki constitutes a significant portion of the soundtrack for Stanley Kubrick’s 1980 film
adaptation of Stephen King’s novel The Shining, while the remainder of the film score was
written for the movie by Wendy Carlos and Rachel Elkind. The use of preexisting music in a
film soundtrack is a topic already explored by numerous film and music scholars, but as a Bartók
specialist I was particularly intrigued by the inclusion of Bartók’s music in a horror film, as a
connection between his music and the plot of The Shining was not clear to me. Subject-Position and Béla Bartók’s Music for Strings, Percussion,
and Celesta (1936) in Stanley Kubrick’s The Shining (1980) Written for his
publisher, Universal Edition, Bartók’s 1937 program notes for Music for Strings, Percussion and
Celesta indicate that the piece was not conceived as a programmatic work and did not contain
intentional elements of horror (Suchoff, 2004: 32). However, unique elements of the work’s third
movement, such as its instrumentation, form, and “night music” texture, (used by Bartók as early
as 1926 to depict natural sounds of the night, here, featuring glissandos, string tremolos,
pentatonic scales, and featuring the celesta, piano and harp), likely led to Kubrick and music
editor Gordon Stainforth’s decision to include it to accompany three scenes in The Shining. The
result is effective enhancement of the drama in the scenes, even though the music was not
originally intended to accompany a film or otherwise tell a story. An ecological approach
provides one method of exploring this issue, and so I determined a subject-position for each
movement of Bartók’s work in accordance with the definition of subject-position established by
film scholar Sheila Johnston and elaborated upon by music scholar Eric F. Clarke. In this paper I
attempt to show that the subject-position of the third movement of Bartók’s Music for Strings as
related to the work’s other movements, is similar to the viewer’s subject-position during the
scenes in The Shining accompanied by excerpts from the movement. pentatonic scales, and featuring the celesta, piano and harp), likely led to Kubrick and music
editor Gordon Stainforth’s decision to include it to accompany three scenes in The Shining. The
result is effective enhancement of the drama in the scenes, even though the music was not
originally intended to accompany a film or otherwise tell a story. An ecological approach
provides one method of exploring this issue, and so I determined a subject-position for each
movement of Bartók’s work in accordance with the definition of subject-position established by
film scholar Sheila Johnston and elaborated upon by music scholar Eric F. Clarke. In this paper I
attempt to show that the subject-position of the third movement of Bartók’s Music for Strings as
related to the work’s other movements, is similar to the viewer’s subject-position during the
scenes in The Shining accompanied by excerpts from the movement. An idea of what is meant by “subject-position” in the analysis of film and music was
essential for undertaking this study. Subject-Position and Béla Bartók’s Music for Strings, Percussion,
and Celesta (1936) in Stanley Kubrick’s The Shining (1980) e-Journal of the American Hungarian Educators
Association, Volume 15 (2022): http://ahea.pitt.edu DOI: 10.5195/ahea.2022.468 attention to the texts of the popular songs and the texture, melody, and harmony in the classical
instrumental pieces. He does not, however, address the formal characteristics of the music, which
play a vital role in analyzing the subject-position of Bartók’s Music for Strings. attention to the texts of the popular songs and the texture, melody, and harmony in the classical
instrumental pieces. He does not, however, address the formal characteristics of the music, which
play a vital role in analyzing the subject-position of Bartók’s Music for Strings. p y
y
g
j
p
f
g
Music for Strings was commissioned on June 23, 1936 by Paul Sacher, founder and
director of the Basel Chamber Orchestra, to commemorate the ensemble’s tenth anniversary. Bartók accepted the commission within four days and completed the composition in three
months. The work’s unique instrumentation was not entirely Bartók’s decision, as Sacher
requested that he write for “a piano or cembalo (as a continuo, so to speak) or some kind of
percussion instrument” in place of winds (Suchoff, 2004: 31). The elimination of winds from the
work was based on technical and monetary difficulties, not an aesthetic idea of Sacher’s
(Suchoff, 2004: 32). Bartók managed, however, to create a unique work within the terms of the
commission, incorporating double string orchestra, celesta, harp, piano, xylophone, timpani, and
one other percussion part covering a variety of instruments. The work is even described by
Bartók’s biographer Halsey Stevens as “three-dimensional” due to the stage organization
dictated by the composer. It is my analysis that formal elements of the music most effectively dictate a subject-
position for each movement. The work is neotonal and highly organized, with a fugue theme
uniting all four movements. The first movement of Music for Strings, Percussion, and Celesta is
a slow fugue in A. As counterpoint is meant to show off music as a material to be manipulated
by a masterful composer, a fugue encourages the listener to adopt the subject-position of a
connoisseur admiring a musical masterpiece. The second movement of the work is a fast sonata
form in C. The classical and somewhat predictable nature of this form puts the listener at ease
and again places him or her outside the music as a distant observer. Subject-Position and Béla Bartók’s Music for Strings, Percussion,
and Celesta (1936) in Stanley Kubrick’s The Shining (1980) In movement three, however,
the listener is immediately pulled into the mysterious soundscape first created by the xylophone
and timpani. No longer an observer, the audience member must devote more attention to the
music in order to make sense of its unfamiliar elements. The separation of the tonal centers of
movements two and three by a tritone further sets the third movement apart from the second. The
movement, a slow arch form (ABCB’A’), is formally a palindrome containing nearly
unrecognizable folk elements and Bartók’s “night music,” a style he first utilized in the fourth
movement of his piano suite entitled Out of Doors (1926) to create a somewhat eerie nocturnal
atmosphere through the use of cluster chords, repeated melodic fragments, and references folk
music. The fugue theme from the first movement is presented transposed and backwards
(retrograde), reminding the listener of the once-familiar texture and melody now fractured in this
alien environment. The listener, however, soon emerges from the arch form of movement three
and back into the subject-position of the observer in movement four, a fast rondo form in A (the
same key as movement I) containing several stylized folk dances. (retrograde), reminding the listener of the once-familiar texture and melody now fractured in this
alien environment. The listener, however, soon emerges from the arch form of movement three
and back into the subject-position of the observer in movement four, a fast rondo form in A (the
same key as movement I) containing several stylized folk dances. y
g
y
Stanley Kubrick’s 1980 film The Shining is a somewhat loose interpretation of Stephen
King’s novel of the same name. A general understanding of the plot is helpful for understanding
the argument presented in this paper. The Shining is the story of Jack Torrence, an out-of-work,
recovering alcoholic and aspiring writer, who accepts a position as the winter caretaker at the
Overlook Hotel in the Rocky Mountains of Colorado despite the fact that the isolation and cabin
fever resulting from being snowed in without guests had driven a previous caretaker to murder
his family and commit suicide. Jack, his wife Wendy and their young son Danny move to the
hotel for the winter months. Before the guests and hotel staff vacate the premises, the family
meets Dick Halloran, a cook who perceives that Danny has psychic powers. Subject-Position and Béla Bartók’s Music for Strings, Percussion,
and Celesta (1936) in Stanley Kubrick’s The Shining (1980) Clarke identifies subject-position in music as “the way in
which characteristics of the musical material shape the general character of a listener’s response
or engagement (involved, repelled, indifferent)” (Clarke, 2005: 91-92). He relates subject-
position to the ecological approach to analyzing music in that a listener’s perception of a musical
work is based on the relationship between “the capacities, sensitivities, and interests of a
perceiver” and the information available to that perceiver in the environment (Clarke, 2005: 91). In his discussion of subject-position, Clarke focuses on the way aesthetic viewpoints are defined
by this relationship. The idea of subject-position is not new but is based on a type of film study. Clarke bases his definition on film writer Sheila Johnston’s explanation of subject-position
(Johnston, 1985: 245). Johnston problematizes the idea of a single possible subject-position,
admitting that there may be as many possible subject-positions as there are viewers. On the one hand, there is the empirical spectator whose interpretation of film will be
determined by all manner of extraneous factors like personal biography, class origins,
previous viewing experience, the variable of conditions of reception etc. On the other
hand, the abstract notion of a ‘subject-position,’ which could be defined as the way in
which a film solicits, demands even, a certain closely circumscribed reading from a
viewer by mean of its own formal operations (Clarke, 2005: 245). Johnston’s definition not only allows for multiple interpretations of a scene based on individual
viewers’ personal experiences, but also insists that the filmmaker has narrowed the range of
possible subject-positions for the viewer to adopt with his or her aesthetic decisions. A music
scholar, Clarke uses these ideas to analyze both popular and classical music, calling special Johnston’s definition not only allows for multiple interpretations of a scene based on individual
viewers’ personal experiences, but also insists that the filmmaker has narrowed the range of
possible subject-positions for the viewer to adopt with his or her aesthetic decisions. A music
scholar, Clarke uses these ideas to analyze both popular and classical music, calling special 147 Lucas, Sarah. “Subject-Position and Béla Bartók’s Music for Strings, Percussion, and Celesta (1936) in Stanley
Kubrick’s The Shining (1980).” Hungarian Cultural Studies. e-Journal of the American Hungarian Educators
Association, Volume 15 (2022): http://ahea.pitt.edu DOI: 10.5195/ahea.2022.468 Lucas, Sarah. “Subject-Position and Béla Bartók’s Music for Strings, Percussion, and Celesta (1936) in Stanley
Kubrick’s The Shining (1980).” Hungarian Cultural Studies. Subject-Position and Béla Bartók’s Music for Strings, Percussion,
and Celesta (1936) in Stanley Kubrick’s The Shining (1980) Halloran calls these
visions “shining” and tells him that he may have visions of terrible things that have happened at 148 Lucas, Sarah. “Subject-Position and Béla Bartók’s Music for Strings, Percussion, and Celesta (1936) in Stanley
Kubrick’s The Shining (1980).” Hungarian Cultural Studies. e-Journal of the American Hungarian Educators
Association, Volume 15 (2022): http://ahea.pitt.edu DOI: 10.5195/ahea.2022.468 Lucas, Sarah. “Subject-Position and Béla Bartók’s Music for Strings, Percussion, and Celesta (1936) in Stanley
Kubrick’s The Shining (1980).” Hungarian Cultural Studies. e-Journal of the American Hungarian Educators
Association, Volume 15 (2022): http://ahea.pitt.edu DOI: 10.5195/ahea.2022.468 the hotel. He also warns Danny not to enter Room 237. After the hotel is empty, Jack quickly
becomes irritable and suffers from writer’s block and insomnia. He experiences hallucinations
and murderous impulses toward Wendy and Danny. He eventually gives into these impulses, but
Wendy and Danny manage to escape, leaving Jack to freeze to death in the hotel’s hedge-maze. The three scenes examined in this study are not only connected by Bartók’s music. While no violence occurs in the scenes featuring the Bartók excerpts, they may be considered
foreshadowing for more disturbing events later in the film in the same locations. The scenes also
impart a sense of association between the audience member and the characters Wendy and
Danny. This is emphasized by the use of the Steadicam, a new technology at the time of the
film’s creation, in the scenes where the audience seems to float in front of or behind the
characters (Code, 2010: 138). The audience is not placed in the subject-position of an evil
presence following Wendy or Danny. The feeling, rather, is that whatever is threatening the
characters in the movie is also threatening the viewer. Each excerpt is from the third movement
of Music for Strings, Percussion, and Celesta and ends with the cymbal crash at m. 45. This
abrupt ending indicates a change of scene and subject-position within the movie. The first
excerpt, which accompanies the maze scene, is the shortest (mm. 20-45), the second, in the scene
in which Danny attempts to enter Room 237 and finds it locked, incorporates only three more
measures (mm. 17-45), and the third, featuring Jack and Danny in the family apartment, utilizes
the entire movement up to m. 45. p
In the first scene accompanied by Bartók’s music, Wendy and Danny are exploring the
hotel’s hedge maze. Subject-Position and Béla Bartók’s Music for Strings, Percussion,
and Celesta (1936) in Stanley Kubrick’s The Shining (1980) They race from the hotel to the maze, and the music begins when they reach
it. Prior to this point the subject-position is that of a far-off observer. When the music starts and
the characters enter the maze, the audience member is immediately pulled inward. Because of the
changing camera position, the observer feels closer to Wendy and Danny, as if floating a few feet
in front of or behind them. Jack, who stays inside the hotel, stares into the model of the maze
while his wife and son are walking through the real version. The subject-position is different at
this point, as the viewer is not invited to adopt a position that associates him or her with Jack in
the way that the Steadicam technique invites the viewer to identify with his wife and son in the
maze. This scene may foreshadow the end of the film, where Danny escapes Jack, who chases
him with an axe in a murderous rage and later freezes to death. In the second scene in which Bartók’s music is utilized, Danny rides through the hotel
hallways on his big wheel. He stops when he notices Room 237 – a place Halloran told him to
avoid – and slowly approaches on foot. When he tries to open the door, he finds it is locked and
“shines” for a moment. He has seen the girls who were killed by their father in the hotel. The
audience is following closely behind Danny as he rides down the halls, yet sees the image of his
shining as if they were inside his mind. The audience is pulled into Danny’s mind just as he is
pulled toward Room 237 by his curiosity. This subject-position is again similar to that of the
third movement of Bartók’s Music for Strings, Percussion, and Celesta. Significant events occur
in Room 237 later in the film: Danny gains access to Room 237 and returns to his parents with a
torn sweater and bruises on his neck. When Jack goes to see what or who may be in the room, he
has a graphic hallucination. The final scene featuring a Bartók excerpt incorporates the longest portion from Music
for Strings, Percussion and Celesta in the film, from the beginning of the third movement
through measure 45. Subject-Position and Béla Bartók’s Music for Strings, Percussion,
and Celesta (1936) in Stanley Kubrick’s The Shining (1980) There is a great deal of coordination between the music and the acting at the
beginning of the scene, which shows Danny attempting to avoid waking his father while entering
the family apartment of the hotel to retrieve a toy. He finds his father awake, sitting on the bed, 149 Lucas, Sarah. “Subject-Position and Béla Bartók’s Music for Strings, Percussion, and Celesta (1936) in Stanley
Kubrick’s The Shining (1980).” Hungarian Cultural Studies. e-Journal of the American Hungarian Educators
Association, Volume 15 (2022): http://ahea.pitt.edu DOI: 10.5195/ahea.2022.468 Lucas, Sarah. “Subject-Position and Béla Bartók’s Music for Strings, Percussion, and Celesta (1936) in Stanley
Kubrick’s The Shining (1980).” Hungarian Cultural Studies. e-Journal of the American Hungarian Educators
Association, Volume 15 (2022): http://ahea.pitt.edu DOI: 10.5195/ahea.2022.468 Lucas, Sarah. “Subject-Position and Béla Bartók’s Music for Strings, Percussion, and Celesta (1936) in Stanley
Kubrick’s The Shining (1980).” Hungarian Cultural Studies. e-Journal of the American Hungarian Educators
Association, Volume 15 (2022): http://ahea.pitt.edu DOI: 10.5195/ahea.2022.468 staring into space. Jack asks Danny to come to him and assures the boy that he would never do
anything to hurt him or his mother. The viewer feels associated with Danny in this scene. As the
audience member watches the door open from inside the apartment, there is a distinct feeling of
danger for both Danny and the viewer. It is (again) the feeling that whatever is threatening
Danny may also be a threat to the observer. Like the others including a Bartók excerpt, this scene
occurs in a location where violence takes place later in the movie. Near the end of the film, Jack
breaks into the apartment and tries to kill his wife and son with an axe. In conclusion, my analysis of the subject-position of Bartók’s Music for Strings, In conclusion, my analysis of the subject-position of Bartók’s Music for Strings,
Percussion and Celesta and the scenes in which excerpts of the work appear in Kubrick’s The
Shining reveals similarities in their potential effect on an audience member. Subject-Position and Béla Bartók’s Music for Strings, Percussion,
and Celesta (1936) in Stanley Kubrick’s The Shining (1980) The first, second,
and fourth movements of the piece place the listener outside the music as an observer, while the
third movement – the only one used in The Shining – pulls the listener inward, due in part to its
unique form and elements of “night music.” This inward motion may also be felt by viewers in
the scenes of the film accompanied by Bartók’s piece as they are drawn into the hedge-maze
with Wendy and Danny, pulled toward the door to Room 237 and into Danny’s mind for his
moment of “shining,” and finally into the family apartment. These scenes are not only connected
by the soundtrack, but also by the fact that violent events later occur in those locations. While the
third movement of Bartók’s Music for Strings, Percussion and Celesta was not intended by the
composer to accompany a horror film or otherwise tell a story, the similarity between the
movement’s subject-position within the context of the larger musical work and the subject-
position of the scenes in The Shining which feature it help explain why the music enhances the
storyline and aids in foreshadowing some of the most frightening events in the film. Lucas, Sarah. “Subject-Position and Béla Bartók’s Music for Strings, Percussion, and Celesta (1936) in Stanley
Kubrick’s The Shining (1980).” Hungarian Cultural Studies. e-Journal of the American Hungarian Educators
Association, Volume 15 (2022): http://ahea.pitt.edu DOI: 10.5195/ahea.2022.468 Works Cited Bartók, Béla. 1939. Music for String Instruments, Percussion, and Celesta (in Four Movements). Boosey & Hawkes. Bernard, Jonathan W. 2003. “Zones of Impingement: Two Movements from Bartók’s Music for
Strings, Percussion and Celesta.” Music Theory Spectrum, vol. 25, no. 1, pp. 3-34. Clarke, Eric F. 2005.Ways of Listening: An Ecological Approach to the Perception of Musical
Meaning. Oxford UP. Code, David J. 2010. “Rehearing The Shining: Musical Undercurrents in the Overlook Hotel.”
Music in the Horror Film: Listening to Fear, edited by Neil Lerner, Routledge, 133-51. Code, David J. 2010. “Rehearing The Shining: Musical Undercurrents in the Overlook Hotel.”
Music in the Horror Film: Listening to Fear, edited by Neil Lerner, Routledge, 133-51. Donnelly, K.J. 2013.“‘Pre’ and ‘Post’ Sound.” Occult Aesthetics: Synchronization in Sound
Film, Oxford UP, 154-180. Music in the Horror Film: Listening to Fear, edited by Neil Lerner, Routledge, 133-51. Donnelly, K.J. 2013.“‘Pre’ and ‘Post’ Sound.” Occult Aesthetics: Synchronization in Sound
Film, Oxford UP, 154-180. Hayward, Philip, ed. 2009. Terror Tracks: Music, Sound and Horror Cinema. Equinox. J
ki
G
1997 S
l
K b i k
d h A
f Ad
i
M F l
d Hayward, Philip, ed. 2009. Terror Tracks: Music, Sound and Horror Cinema. Equi kins, Greg. 1997. Stanley Kubrick and the Art of Adaptation. McFarland. Johnston, Sheila. 1985. “Film Narrative and the Structuralist Controversy.” The Cinema Book,
edited by P. Cook, British Film Institute, 221-50. Lutz, John. 2010. “From Domestic Nightmares to the Nightmare of History: Uncanny Eruptions
of Violence in King’s and Kubrick’s Versions of The Shining.” The Philosophy of
Horror, edited by Thomas Fahy, The University Press of Kentucky, 161-78. Magistrale, Anthony, ed. 1991. The Shining Reader. Starmont House. McQuiston, Kate. 2013. “Mysterious Music with Invisible Edges and the Emergence of Musical
Form in The Shining.” We’ll Meet Again: Musical Design in the Films of Stanley
Kubrick, Oxford UP, 65-86. 150 Palmer, R. Barton. 2007. “The Shining and Anti-Nostalgia: Postmodern Notions of History.” The
Philosophy of Stanley Kubrick, edited Jerold J. Abrams, The University of Kentucky
Press, 201-218. Palmer, R. Barton. 2007. “The Shining and Anti-Nostalgia: Postmodern Notions of History.” The
Philosophy of Stanley Kubrick, edited Jerold J. Abrams, The University of Kentucky
Press, 201-218. The Shining. 1980. Directed by Stanley Kubrick. Warner Home Video, 1999. DVD The Shining. 1980. Directed by Stanley Kubrick. Warner Home Video, 1999. DVD. Somfai, László. 2003. Kubrick s The Shining (1980). Hungarian Cultural Studies. e Journal of the Amer
Association, Volume 15 (2022): http://ahea.pitt.edu DOI: 10.5195/ahea.2022.468 Works Cited “Invention, Form, Narrative in Béla Bartók’s Music.” Studia
Musicologica Acadmiae Scientiarum Hungaricae, vol. 44, nos. 3-4, 291-303. rd g
y
y
,
Somfai, László. 2003. “Invention, Form, Narrative in Béla Bartók’s Music.” Studia
Musicologica Acadmiae Scientiarum Hungaricae, vol. 44, nos. 3-4, 291-303. Somfai, László. 2003. “Invention, Form, Narrative in Béla Bartók’s Music.” Studia
Musicologica Acadmiae Scientiarum Hungaricae, vol. 44, nos. 3-4, 291-303
d Stevens, Halsey. 1993. The Life and Music of Béla Bartók. 3rd ed. Edited by Malcolm Gillies. Clarendon. Suchoff, Benjamin. 2004. Béla Bartók: A Celebration. Scarecrow Press. Vikárius, László. 2006. “Bartók's Late Adventures with ‘Kontrapunkt’.” Studia Musicologica
Academiae Scientiarum Hungaricae, vol. 47, nos. 3-4, Bartók's Orbit. The Context
and Sphere of Influence of His Work. Proceedings of the International Conference
Held by the Bartók Archives, Budapest (22-24 March 2006). Part I, 395-416. Webster, Patrick. 2011. “The Shining: The Misuses of Enchantment.” Love and Death in
Kubrick: A Critical Study of the Films from Lolita through Eyes Wide Shut, McFarland,
88-116. Wright, Jarrell D. 2011. “Shades of Horror: Fidelity and Genre in Stanley Kubrick’s The
Shining.” True to the Spirit: Film Adaptation and the Question of Fidelity, edited by
Colin Maccabe, Kathleen Murray, and Rick Warner, Oxford UP, 173-93. 151
|
https://openalex.org/W2003752678
|
https://europepmc.org/articles/pmc2565666?pdf=render
|
English
| null |
The KBG syndrome: Case report
|
Cases journal
| 2,008
|
cc-by
| 1,777
|
Cases Journal Open Access BioMed Central BioMed Central Case Report
The KBG syndrome: Case report
Ilaria Morghen* and Enrico Ferri Received: 21 May 2008
Accepted: 26 September 2008 Published: 26 September 2008 g
;
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0),
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Introduction: The KBG syndrome is a rare autosomal dominant condition, first described by
Hermann et al. in 1975. Fundamental findings are: mild development delay, short stature,
craniofacial dysmorphism and skeletal anomalies. Case presentation: A 32 years old woman, Caucasian race, weight 57 Kg, affected by KBG
syndrome was sent to our clinics for preoperative anaesthesia evaluation. She was schedules for
left ossicular reconstruction under general anaesthesia for bilateral hearing loss. A psycho-motor
retardation was associated to morphological anomalies such as short neck, hyperlordosis without
neck extension imparirment, craniofacial anomalies and dento-skeletal abnormalities. An
echocardiography showed the presence of interatrial defect with left-to-right shunt. The patient
was sent to a cardiac surgery centre. Conclusion: Perioperative evaluation of patients affected by KBG syndrome must take into
consideration the management of difficult airways, due to the associated craniofacial dysmorphism. The possible presence of cardiac anomalies in the KBG syndrome is currently being evalueted. In
this report the finding of cardiomegaly and congestion of the pulmonary circulation was attributed
to presence of an interatrial defect with left-to-right shunt. The risk of cardiopulmonary failure led
us to ask for a cardiac surgery consult. Perioperative management of these patients should be
extremely accurate, even in the case of minor surgery, and should include also chest X-rays and
echocardiography evaluation. hearing loss for bilateral recurrent otitis media. Skeletal
anomalies can include: cervical ribs, hip dysplasia, thora-
columbar scoliosis, anomalies of hands, femoral and ver-
tebral bones [2-4]. Introduction The KBG syndrome is a rare autosomal dominant condic-
tion, first described by Hermann et al. in 1975. KBG are
the initials of the first patient described whit this syn-
drome [1]. We report a case of KBG syndrome being evaluated for
ossicular reconstruction under general anesthesia, that
was redirected to a cardiac surgery centre. Fundamental findings are: mild developmental delay,
short stature, craniofacial dysmorphism with brachyceph-
aly, round face, hypertelorism, palpebral fissures in the
mongoloid position, macrodontia of the maxillary per-
manent central incisors, hypodontia, short alveolar ridge, Page 1 of 3
(page number not for citation purposes) Page 1 of 3
(page number not for citation purposes) http://www.casesjournal.com/content/1/1/186 http://www.casesjournal.com/content/1/1/186 Cases Journal 2008, 1:186 Authors' contributions Perioperative evaluation of patients affected by KBG must
take into consideration the management of difficult air-
ways, due to the associated craniofacial dysmorphism. In
this case, problems arose from the extreme flattnes of face,
Angle class III malocclusion, short neck with hyperlordo-
sis and macrodontia of the upper central incisors. High
Mallampati score (Type 3) were assessed. MI has performed the preoperative evaluation, conceived
the paper, searched the literature for other similar reports
and review on the subject, drafted the manuscript. FE has
been involved in the analysis of data and helped to draft
the manuscript. All authors read and approved the final
manuscript. p
g
The authors declare that they have no competing interests. p
g
The authors declare that they have no competing interests. Abbreviations
IAD i
i l d IAD: interatrial defect; PAP: pulmonary artery pressure. No left ventricle anomalies were found. Therefore the
patient was sent to a cardiac surgery centre. Conclusion Patients affected by KBG syndrome can present many
challenges to the anaesthesiologist that performs the pre-
operative evaluation. This addresses the issue of consider-
ing the possible association of evolving cardiac anomalies
when dealing with the KBG syndrome. Perioperative man-
agement of these patients should be extremely accurate,
even in the case of minor surgery, and should include also
chest X-rays and echocardiography evaluation. Pre-operative evaluation with an echocardiogram showed
the presence of biatrial enlargment, interatrial defect
(IAD) with left-to-right shunt, right ventricle enlargment,
estimated pulmonary artery pressure (PAP) 45–50
mmHg. Electrocardiogram showed a right bundle branch block. Case presentation Atrial septal defect occurs as an isolated anomaly in 5% to
10% of all congenital heart defects. It is more common in
females than in males and can occur in any portion of the
atrial septum. Right axis deviation and mild right ven-
tricular hypertrophy or right bundle branch block are typ-
ical findings on electrocardiogram. Cardiomegaly with
enlargment of the right atrium and right ventricle may be
present. A 2-dimensional echocardiography is diagnostic
[8]. A 32 years old woman, Caucasian race, weight 57 Kg,
affected by KBG syndrome was sent to our clinics for a pre-
operative anesthesia evaluation. She was scheduled for
left ossicular reconstruction under general anesthesia for
bilateral hearing loss. The patient was born after a normal
uncomplicated pregnancy. Medical history: operation of congenital luxation of the
hip to 19 years in spinal anesthesia. The finding of cardiomegaly and congestion of the pul-
monary circulation was attributed to presence of an IAD
with a functional significance of the left-to-right shunt. The risk of cardiopulmonary failure led us to ask for a car-
diac surgery consult. Upon agreement with the head and
neck surgeon, we decided to postpone the ossiculoplasti
intervention, this being not urgent for the patient, who
carried an acoustic prothesis for bilateral mixed hearing
loss since the age of 8. Although she had always been fol-
lowed from a specialized centre for paediatric diseases,
such a severe cardiac anomaly, undoubtably congenital
and worsened during time, has been detected very late. Clinical examination showed a psycho-motor retardation
associated to morphological anomalies, such as short stat-
ure, short neck, hypelordosis without neck extension
impairment, mild hyperterolism, slight antimongoloid,
extreme flattening of the face, arhinencephalic profile
with right-convex deflection of the nasal septum, macro-
dontia of the upper central incisor, 1.1 and 2.1, hypodon-
tia, anterior inverted bite (Mallampati class 3). Thoracic examination revealed a 2/6 systolic cardiac mur-
mur at the apex. Chest X-ray showed cardiomegaly, signes
of congestion in the pulmonary circulation and increased
hilar shapes. Since many years the patient had been
reporting dispnoea with every airways inflammation
process occurring in the past years. Competing interests p
g
The authors declare that they have no competing interests. Consent The possible presence of cardiac anomalies in the KBG
syndrome is currently being evaluated. At present a single
case of interventricular septum defect, two cases of bicus-
pid aortic valve and partial atrioventricular canal defect,
one stenosis of the left pulmonary artery and a ventricular
septal defect, has been reported [5-7]. Written consent was obtained from the patient for pubbli-
cation of this case report and any accompanying images. A copy of the written consent is avaible for review by the
Editor-in-Chief of this journal. Page 2 of 3
(page number not for citation purposes) Page 2 of 3
(page number not for citation purposes) Cases Journal 2008, 1:186 http://www.casesjournal.com/content/1/1/186 http://www.casesjournal.com/content/1/1/186 References References
1. Dowling PA, Fleming P, Gorlin RJ, King M, Nevin NC, McEntargart :
The KBG syndrome, characteristic dental findings: a case
report. International Journal of Pediatric Dentistry 2001, 11:131-134. 2. Zollino M, Battaglia A, D'Avanzo MG, Della Bruna MM, Marini G,
Scarano G, Cappa M: Six additional cases of the KBG syn-
drome: clinical reports and outline of the diagnostic criteria. Am J Med Genet 1994, 52:302-307. 3. Soekarman D, Volcke P, Fryns JP: The KBG syndrome: follow-up
data on three affected brothers. Clin Genet 1994, 46:283-286. 4. Fryns JP, Haspeslag M: Mental retardation, short stature, minor
skeletal anomalies, craniofacial dysmorphism and macro-
dontia in two sisters and their mother: another variant
example of the KBG syndrome? Clin Genet 1984, 26:69-72. 5. Devriendt K, Holvoet M, Fryns JP: Further delineation of the
KBG syndrome [abstract]. Genet Couns 1998, 9(3):191-194. 6. Brancati F, Sarkozy A, Dallapiccola B: KBG syndrome. Orphanet J
Rare Dis 2006, 1:50. 7. Brancati F, D'Avanzo MG, Digilio MC, Sarzosky A, Biondi M, De Brasi
D, Mingarelli R, Dallapiccola B: KBG Syndrome in a Cohort of
Italian Patients. Am J Med Genet 2004, 131:144-149. 8. Dharam M, Jason ZQ, Vipin M: Congenital heart disease with
left-to-right shunts. International Anestesiology Clinics 2004,
42(4):45-58. 1. Dowling PA, Fleming P, Gorlin RJ, King M, Nevin NC, McEntargart :
The KBG syndrome, characteristic dental findings: a case
report. International Journal of Pediatric Dentistry 2001, 11:131-134. 1. Dowling PA, Fleming P, Gorlin RJ, King M, Nevin NC, McEntargart :
The KBG syndrome, characteristic dental findings: a case
report. International Journal of Pediatric Dentistry 2001, 11:131-134. p
J
y
2. Zollino M, Battaglia A, D'Avanzo MG, Della Bruna MM, Marini G,
Scarano G, Cappa M: Six additional cases of the KBG syn-
drome: clinical reports and outline of the diagnostic criteria. Am J Med Genet 1994, 52:302-307. J
,
3. Soekarman D, Volcke P, Fryns JP: The KBG syndrome: follow-up
data on three affected brothers. Clin Genet 1994, 46:283-286. J
,
3. Soekarman D, Volcke P, Fryns JP: The KBG syndrome: follow-up
data on three affected brothers. Clin Genet 1994, 46:283-286. data on three affected brothers. Clin Genet 1994, 46:283-286. 4. Fryns JP, Haspeslag M: Mental retardation, short stature, minor
skeletal anomalies, craniofacial dysmorphism and macro-
dontia in two sisters and their mother: another variant
example of the KBG syndrome? Clin Genet 1984, 26:69-72. p
y
5. References Devriendt K, Holvoet M, Fryns JP: Further delineation of the
KBG syndrome [abstract]. Genet Couns 1998, 9(3):191-194. 6. Brancati F, Sarkozy A, Dallapiccola B: KBG syndrome. Orphanet J
Rare Dis 2006, 1:50. p
y
5. Devriendt K, Holvoet M, Fryns JP: Further delineation of the
KBG syndrome [abstract]. Genet Couns 1998, 9(3):191-194. ( )
6. Brancati F, Sarkozy A, Dallapiccola B: KBG syndrome. Orphanet J
Rare Dis 2006, 1:50. 7. Brancati F, D'Avanzo MG, Digilio MC, Sarzosky A, Biondi M, De Brasi
D, Mingarelli R, Dallapiccola B: KBG Syndrome in a Cohort of
Italian Patients. Am J Med Genet 2004, 131:144-149. J
8. Dharam M, Jason ZQ, Vipin M: Congenital heart disease with
left-to-right shunts. International Anestesiology Clinics 2004,
42(4):45-58. Publish with BioMed Central and every
scientist can read your work free of charge
"BioMed Central will be the most significant development for
disseminating the results of biomedical research in our lifetime."
Sir Paul Nurse, Cancer Research UK
Your research papers will be:
available free of charge to the entire biomedical community
peer reviewed and published immediately upon acceptance
cited in PubMed and archived on PubMed Central
yours — you keep the copyright
Submit your manuscript here:
http://www.biomedcentral.com/info/publishing_adv.asp
BioMedcentral
Page 3 of 3
(page number not for citation purposes) Publish with BioMed Central and every
scientist can read your work free of charge
|
https://openalex.org/W4247981280
|
https://figshare.com/articles/preprint/SNE-RoadSeg_Incorporating_Surface_Normal_Information_into_Semantic_Segmentation_for_Accurate_Freespace_Detection/12864287/1/files/24451445.pdf
|
English
| null |
SNE-RoadSeg: Incorporating Surface Normal Information into Semantic Segmentation for Accurate Freespace Detection
| null | 2,020
|
cc-by
| 7,170
|
This paper was downloaded from TechRxiv (https://www.techrxiv.org). LICENSE SUBMISSION DATE / POSTED DATE
26-08-2020 / 01-09-2020 SUBMISSION DATE / POSTED DATE
26-08-2020 / 01-09-2020 SNE-RoadSeg: Incorporating Surface Normal Information into
Semantic Segmentation for Accurate Freespace Detection This paper was downloaded from TechRxiv (https://www.techrxiv.org). CITATION Fan, Rui; Wang, Hengli; Cai, Peide; Liu, Ming (2020): SNE-RoadSeg: Incorporating Surface Normal
Information into Semantic Segmentation for Accurate Freespace Detection. TechRxiv. Preprint. https://doi.org/10.36227/techrxiv.12864287.v1 10.36227/techrxiv.12864287.v1 10.36227/techrxiv.12864287.v1 SNE-RoadSeg: Incorporating Surface Normal
Information into Semantic Segmentation for
Accurate Freespace Detection Rui Fan1⋆[0000−0003−2593−6596], Hengli Wang2⋆[0000−0002−7515−9759],
Peide Cai2[0000−0002−9759−2991], and Ming Liu2[0000−0002−4500−238X] Rui Fan1⋆[0000−0003−2593−6596], Hengli Wang2⋆[0000−0002−7515−9759],
Peide Cai2[0000−0002−9759−2991], and Ming Liu2[0000−0002−4500−238X] Rui Fan1⋆[0000−0003−2593−6596], Hengli Wang2⋆[0000−0002−7515−9759],
Peide Cai2[0000−0002−9759−2991], and Ming Liu2[0000−0002−4500−238X] 1 UC San Diego
rui.fan@ieee.org
2 HKUST Robotics Institute
} 1 UC San Diego
rui.fan@ieee.org
2 HKUST Robotics Institute
{hwangdf, peide.cai, eelium}@ust.hk 1 UC San Diego
rui.fan@ieee.org
2 HKUST Robotics Institute
{hwangdf, peide.cai, eelium}@ust.hk {hwangdf, peide.cai, eelium}@ust.hk Abstract. Freespace detection is an essential component of visual per-
ception for self-driving cars. The recent efforts made in data-fusion con-
volutional neural networks (CNNs) have significantly improved semantic
driving scene segmentation. Freespace can be hypothesized as a ground
plane, on which the points have similar surface normals. Hence, in this
paper, we first introduce a novel module, named surface normal esti-
mator (SNE), which can infer surface normal information from dense
depth/disparity images with high accuracy and efficiency. Furthermore,
we propose a data-fusion CNN architecture, referred to as RoadSeg,
which can extract and fuse features from both RGB images and the
inferred surface normal information for accurate freespace detection. For
research purposes, we publish a large-scale synthetic freespace detec-
tion dataset, named Ready-to-Drive (R2D) road dataset, collected under
different illumination and weather conditions. The experimental results
demonstrate that our proposed SNE module can benefit all the state-
of-the-art CNNs for freespace detection, and our SNE-RoadSeg achieves
the best overall performance among different datasets. Keywords: freespace detection · self-driving cars · data-fusion CNN
· semantic driving scene segmentation · surface normal Source Code, Dataset and Demo Video:
sites.google.com/view/sne-roadseg ⋆These authors contributed equally to this work and are therefore joint first authors. 1
Introduction Autonomous cars are a regular feature in science fiction films and series, but
thanks to the rise of artificial intelligence, the fantasy of picking up one such
vehicle at your garage forecourt has turned into reality. Driving scene under-
standing is a crucial task for autonomous cars, and it has taken a big leap ⋆These authors contributed equally to this work and are therefore joint first authors. R. Fan, H. Wang, P. Cai, and M. Liu 2 with recent advances in artificial intelligence [1]. Collision-free space (or simply
freespace) detection is a fundamental component of driving scene understanding
[27]. Freespace detection approaches generally classify each pixel in an RGB or
depth/disparity image as drivable or undrivable. Such pixel-level classification
results are then utilized by other modules in the autonomous system, such as
trajectory prediction [4] and path planning [31], to ensure that the autonomous
car can navigate safely in complex environments. The existing freespace detection approaches can be categorized as either tra-
ditional or machine/deep learning-based. The traditional approaches generally
formulate freespace with an explicit geometry model and find its best coefficients
using optimization approaches [13]. [36] is a typical traditional freespace detec-
tion algorithm, where road segmentation is performed by fitting a B-spline model
to the road disparity projections on a 2D disparity histogram (generally known
as a v-disparity image) [12]. With recent advances in machine/deep learning,
freespace detection is typically regarded as a semantic driving scene segmenta-
tion problem, where the convolutional neural networks (CNNs) are used to learn
its best solution [34]. For instance, Lu et al. [25] employed an encoder-decoder ar-
chitecture to segment RGB images in the bird’s eye view for end-to-end freespace
detection. Recently, many researchers have resorted to data-fusion CNN archi-
tectures to further improve the accuracy of semantic image segmentation. For
example, Hazirbas et al. [19] incorporated depth information into conventional
semantic segmentation via a data-fusion CNN architecture, which greatly en-
hanced the performance of driving scene segmentation.i In this paper, we first introduce a novel module named surface normal es-
timator (SNE), which can infer surface normal information from dense dispar-
ity/depth images with both high precision and efficiency. Additionally, we de-
sign a data-fusion CNN architecture named RoadSeg, which is capable of in-
corporating both RGB and surface normal information into semantic segmen-
tation for accurate freespace detection. 3 cvlibs.net/datasets/kitti/eval_road.php 2
Related Work In 2015, Long et al. [24] introduced Fully Convolutional Network (FCN), a CNN
for end-to-end semantic image segmentation. Since then, research on this topic
has exploded. Based on FCN, Ronneberger et al. [26] proposed U-Net in the
same year, which consists of a contracting path and an expansive path [26]. It adds skip connections between the contracting path and the expansive path
to help better recover the full spatial resolution. Different from FCN, SegNet
[3] utilizes an encoder-decoder architecture, which has become the mainstream
structure for following approaches. An encoder-decoder architecture is typically
composed of an encoder, a decoder and a final pixel-wise classification layer. Furthermore, DeepLabv3+ [9], developed from DeepLabv1 [6], DeepLabv2 [7]
and DeepLabv3 [8], was proposed in 2018. It employs depth-wise separable con-
volution in both atrous spatial pyramid pooling (ASPP) and the decoder, which
makes its encoder-decoder architecture much faster and stronger [9]. Although
the ASPP can generate feature maps by concatenating multiple atrous-convolved
features, the resolution of the generated feature maps is not sufficiently dense
for some applications such as autonomous driving [7]. To address this problem,
DenseASPP [37] was designed to connect atrous convolutional layers (ACLs)
densely. It is capable of generating multi-scale features that cover a larger and
denser scale range, without significantly increasing the model size [37].f Different from the above-mentioned CNNs, DUpsampling [32] was proposed
to recover the pixel-wise prediction by employing a data-dependent decoder. It allows the decoder to downsample the fused features before merging them,
which not only reduces computational costs, but also decouples the resolutions
of both the fused features and the final prediction [32]. GSCNN [30] utilizes a
novel two-branch architecture consisting of a regular (classical) branch and a
shape branch. The regular branch can be any backbone architecture, while the
shape branch processes the shape information in parallel with the regular branch. Experimental results have demonstrated that this architecture can significantly
boost the performance on thinner and smaller objects [30]. FuseNet [19] was designed to use RGB-D data for semantic image segmenta-
tion. The key ingredient of FuseNet is a fusion block, which employs element-wise
summation to combine the feature maps obtained from two encoders. Although
FuseNet [19] demonstrates impressive performance, the ability of CNNs to handle
geometric information is limited, due to the fixed grid kernel structure [35]. 1
Introduction Since the existing freespace detection
datasets with diverse illumination and weather conditions do not have either
disparity/depth information or freespace ground truth, we created a large-scale
synthetic freespace detection dataset, named Ready-to-Drive (R2D) road dataset
(containing 11430 pairs of RGB and depth images), under different illumination
and weather conditions. Our R2D road dataset is also publicly available for re-
search purposes. To validate the feasibility and effectiveness of our introduced
SNE module, we use three road datasets (KITTI [15], SYNTHIA [21] and our
R2D) to train ten state-of-the-art CNNs (six single-modal CNNs and four data-
fusion CNNs), with and without our proposed SNE module embedded. The
experiments demonstrate that our proposed SNE module can benefit all these
CNNs for freespace detection. Also, our method SNE-RoadSeg outperforms all
other CNNs for freespace detection, where its overall performance is the second
best on the KITTI road benchmark3 [15]. The remainder of this paper is structured as follows: Section 2 provides an
overview of the state-of-the-art CNNs for semantic image segmentation. Section 3 cvlibs.net/datasets/kitti/eval_road.php SNE-RoadSeg 3 3 3 introduces our proposed SNE-RoadSeg. Section 4 shows the experimental re-
sults and discusses both the effectiveness of our proposed SNE module and the
performance of our SNE-RoadSeg. Finally, Section 5 concludes the paper. 3 introduces our proposed SNE-RoadSeg. Section 4 shows the experimental re-
sults and discusses both the effectiveness of our proposed SNE module and the
performance of our SNE-RoadSeg. Finally, Section 5 concludes the paper. 2
Related Work To
address this problem, depth-aware CNN [35] presents two intuitive and flexible
operations: depth-aware convolution and depth-aware average pooling. These
operations can efficiently incorporate geometric information into the CNN by
leveraging the depth similarity between pixels [35]. R. Fan, H. Wang, P. Cai, and M. Liu 4 4 1
2
3
4
4, 0
3, 0
2, 0
3, 1
2, 1
2, 2
1, 0
1, 1
1, 2
1, 3
1
2
3
4
1, 1
1, 2
1, 3
2, 1
2, 2
1, 0
2, 0
3, 0
3, 1
4, 0
+
+
+
+
+
𝑐𝑐0
𝑠𝑠/2
𝑐𝑐0
𝑐𝑐1
𝑠𝑠/4
𝑐𝑐1
𝑐𝑐2
𝑠𝑠/8
𝑐𝑐2
𝑐𝑐3
𝑠𝑠/16
𝑐𝑐3
𝑐𝑐4
𝑠𝑠/32
𝑐𝑐4
Depth Image
RGB Image
Semantic Prediction
SNE
0, 0
𝑠𝑠
Data Flow
Convolution
Batch Normalization
ReLU
+ Element-Wise Summation
c
Max Pooling
Sigmoid
Fusion Layer
𝒏𝒏
Concatenation
The 𝑛𝑛-th
Residual Layer
Upsampling
Layer 𝑈𝑈𝑖𝑖,𝑗𝑗
𝒊𝒊, 𝒋𝒋
Feature
Extractor 𝐹𝐹𝑖𝑖,𝑗𝑗
𝒊𝒊, 𝒋𝒋
c
𝒊𝒊, 𝒋𝒋
𝒊𝒊, 𝒋𝒋
Upsampling Layer 𝑈𝑈𝑖𝑖,𝑗𝑗
Bi-linear
Upsampling
𝑠𝑠/2𝑖𝑖
𝑠𝑠/2𝑖𝑖
𝑐𝑐𝑖𝑖
Feature Extractor 𝐹𝐹𝑖𝑖,𝑗𝑗
⋮
𝑠𝑠/2𝑖𝑖
𝑐𝑐𝑖𝑖−1
𝑠𝑠/2𝑖𝑖
𝑐𝑐𝑖𝑖−1
𝑠𝑠/2𝑖𝑖
(𝑗𝑗+ 2)𝑐𝑐𝑖𝑖−1
ቊ2 , 𝑖𝑖= 0
𝑐𝑐𝑖𝑖−1, 𝑖𝑖> 0
𝑠𝑠/2𝑖𝑖+1
𝑐𝑐𝑖𝑖
Fig. 1: The architecture of our SNE-RoadSeg. It consists of our SNE module, an
RGB encoder, a surface normal encoder and a decoder with densely-connected
skip connections. s represents the input resolution of the RGB and depth images. cn represents the number of feature map channels at different levels. RGB Image Fig. 1: The architecture of our SNE-RoadSeg. It consists of our SNE module, an
RGB encoder, a surface normal encoder and a decoder with densely-connected
skip connections. s represents the input resolution of the RGB and depth images. cn represents the number of feature map channels at different levels. MFNet [18] was proposed for semantic driving scene segmentation with the
use of RGB-thermal vision data. In order to meet the real-time requirement of
autonomous driving applications, MFNet focuses on minimizing the trade-off be-
tween accuracy and efficiency. Similarly, RTFNet [29] was developed to improve
the semantic image segmentation performance using RGB-thermal vision data. Its main contribution is a novel decoder, which leverages short-cuts to produce
sharp boundaries while keeping more detailed information [29]. 3.1
SNE The proposed SNE is developed from our recent work three-filters-to-normal
(3F2N) [14]. Its architecture is shown in Fig. 2. For a perspective camera model,
a 3D point P = [X,Y, Z]⊤in the Euclidean coordinate system can be linked with
a 2D image pixel p = [x, y]⊤using: Z
[p
1
]
= KP =
fx 0 xo
0 fy yo
0 0 1
P,
(1) (1) where K is the camera intrinsic matrix; po = [xo, yo]⊤is the image center; fx
and fy are the camera focal lengths in pixels. The simplest way to estimate the
surface normal n = [nx, ny, nz]⊤of P is to fit a local plane: where K is the camera intrinsic matrix; po = [xo, yo]⊤is the image center; fx
and fy are the camera focal lengths in pixels. The simplest way to estimate the
surface normal n = [nx, ny, nz]⊤of P is to fit a local plane: nxX + nyY + nzZ + d = 0
(2) (2) SNE-RoadSeg 5 Fig. 2: The architecture of our proposed SNE module. Fig. 2: The architecture of our proposed SNE module. to N+
P = [P, NP]⊤, where NP = [Q1, . . ., Qk]⊤is a set of k neighboring points of
P. Combining (1) and (2) results in [14]: to N+
P = [P, NP]⊤, where NP = [Q1, . . ., Qk]⊤is a set of k neighboring points of
P. Combining (1) and (2) results in [14]: 1
Z = −1
d
(
nx
x −xo
fx
+ ny
y −yo
fy
+ nz
)
. (3) (3) Differentiating (3) with respect to x and y leads to: Differentiating (3) with respect to x and y leads to: Differentiating (3) with respect to x and y leads to: gx = ∂1/Z
∂x
= −nx
dfx
,
gy = ∂1/Z
∂y
= −ny
dfy
,
(4) (4) which, as illustrated in Fig. 2, can be respectively approximated by convolving
the inverse depth image 1/Z (or a disparity image, as disparity is in inverse
proportion to depth) with a horizontal and a vertical image gradient filter [14]. Rearranging (4) results in the expressions of nx and ny as follows: which, as illustrated in Fig. 3.1
SNE ˆn can therefore be estimated by mini-
mizing E = −∑k
i=1 ˆn · ¯ni [13]. ∂E
∂θ = 0 obtains: θ = arctan
(∑k
i=1 ¯nxi cos φ + ∑k
i=1 ¯nyi sin φ
∑k
i=1 ¯nzi
)
. (10) (10) Substituting θ and φ into (8) results in the optimal surface normal ˆn, as shown
in Fig. 2. The performance of our proposed SNE will be discussed in Section 4. Substituting θ and φ into (8) results in the optimal surface normal ˆn, as shown
in Fig. 2. The performance of our proposed SNE will be discussed in Section 4. 3.1
SNE 2, can be respectively approximated by convolving
the inverse depth image 1/Z (or a disparity image, as disparity is in inverse
proportion to depth) with a horizontal and a vertical image gradient filter [14]. Rearranging (4) results in the expressions of nx and ny as follows: nx = −dfxgx,
ny = −dfygy. (5) (5) Given an arbitrary Qi ∈NP, we can compute its corresponding nzi by plugging
(5) into (2): (5) nzi = d fx∆Xigx + fy∆Yigy
∆Zi
,
(6) (6) where Qi −P = [∆Xi, ∆Yi, ∆Zi]⊤. Since (5) and (6) have a common factor of −d,
the surface normal ni obtained from Qi and P has the following expression [34]: where Qi −P = [∆Xi, ∆Yi, ∆Zi]⊤. Since (5) and (6) have a common factor of −d,
the surface normal ni obtained from Qi and P has the following expression [34]: ni =
[
fxgx,
fygy,
−fx∆Xigx + fy∆Yigy
∆Zi
]⊤
. (7) (7) A k-connected neighborhood system NP of P can produce k normalized surface
normals ¯n1, …, ¯nk, where ¯ni =
ni
∥ni ∥2 = [¯nxi, ¯nyi, ¯nzi]⊤. Since any normalized surface
normals are projected on a sphere with center (0, 0, 0) and radius 1, we believe
that the optimal surface normal ˆn for P is also projected somewhere on the same
sphere, where the projections of ¯n1, …, ¯nk distribute most intensively [13]. ˆn can
be written in spherical coordinates as follows: ˆn =
[
sin θ cos φ, sin θ sin φ, cos θ
]⊤
,
(8) (8) R. Fan, H. Wang, P. Cai, and M. Liu R. Fan, H. Wang, P. Cai, and M. Liu R. Fan, H. Wang, P. Cai, and M. Liu 6 where θ ∈[0, π] denotes inclination and φ ∈[0, 2π) denotes azimuth. φ can be
computed using: where θ ∈[0, π] denotes inclination and φ ∈[0, 2π) denotes azimuth. φ can be
computed using: φ = arctan
( fygy
fxgx
)
. (9) (9) Similar to [13], we hypothesize that the angle between an arbitrary pair of nor-
malized surface normals is less than π/2. ˆn can therefore be estimated by mini-
mizing E = −∑k
i=1 ˆn · ¯ni [13]. ∂E
∂θ = 0 obtains: Similar to [13], we hypothesize that the angle between an arbitrary pair of nor-
malized surface normals is less than π/2. 4.1
Datasets and Experimental Setup In our experiments, we first evaluate the performance of our proposed SNE on
the DIODE dataset [33], a public surface normal estimation dataset containing
RGBD vision data of both indoor and outdoor scenarios. We utilize the average
angular error (AAE), eAAE = 1
m
∑m
k=1 cos−1 (
⟨nk,ˆnk ⟩
∥nk ∥2 ∥ˆnk ∥2
)
, to quantify our SNE’s
accuracy, where m is the number of 3D points used for evaluation; nk and ˆnk
is the ground truth and estimated (optimal) surface normal, respectively. The
experimental results are presented in Section 4.2. Then, we carry out the experiments on the following three datasets to eval-
uate the performance of our proposed SNE-RoadSeg for freespace detection: – The KITTI road dataset [15]: this dataset provides real-world RGB-D vision
data. We split it into three subsets: a) training (173 images), b) validation
(58 images), and c) testing (58 images). – The KITTI road dataset [15]: this dataset provides real-world RGB-D vision
data. We split it into three subsets: a) training (173 images), b) validation
(58 images), and c) testing (58 images). – The SYNTHIA road dataset [21]: this dataset provides synthetic RGB-D
vision data. We select 2224 images from it and group them into: a) training
(1334 images), b) validation (445 images), and c) testing (445 images). – Our R2D road dataset: along with our proposed SNE-RoadSeg, we also pub-
lish a large-scale synthetic freespace detection dataset, named R2D road
dataset. This dataset is created using the CARLA4 simulator [11]. Firstly,
we mount a simulated stereo rig (baseline: 1.5 m) on the top of a vehicle to
capture synchronized stereo images (resolution: 640×480 pixels) at 10 fps. The vehicle navigates in six different scenarios under different illumination
and weather conditions (sunny, rainy, day and sunset). There are a total of
11430 pairs of stereo images with corresponding depth images and semantic
segmentation ground truth. We split them into three subsets: a) training
(6117 images), b) validation (2624 images), and c) testing (2689 images). Our dataset is publicly available at sites.google.com/view/sne-roadseg
for research purposes. We use these three datasets to train ten state-of-the-art CNNs, including six
single-modal CNNs and four data-fusion CNNs. 3.2
RoadSeg U-Net [26] has demonstrated the effectiveness of using skip connections in re-
covering the full spatial resolution. However, its skip connections force aggrega-
tions only at the same-scale feature maps of the encoder and decoder, which,
we believe, is an unnecessary constraint. Inspired by DenseNet [23], we propose
RoadSeg, which exploits densely-connected skip connections to realize flexible
feature fusion in the decoder. As shown in Fig. 1, our proposed RoadSeg also adopts the popular encoder-
decoder architecture. An RGB encoder and a surface normal encoder is employed
to extract the feature maps from RGB images and from the inferred surface nor-
mal information, respectively. The extracted RGB and surface normal feature
maps are hierarchically fused through element-wise summations. The fused fea-
ture maps are then fused again in the decoder through densely-connected skip
connections to restore the resolution of the feature maps. At the end of RoadSeg,
a sigmoid layer is used to generate the probability map for the semantic driving
scene segmentation. We use ResNet [20] as the backbone of our RGB and surface normal encoders,
the structures of which are identical to each other. Specifically, the initial block
consists of a convolutional layer, a batch normalization layer and a ReLU acti-
vation layer. Then, a max pooling layer and four residual layers are sequentially
employed to gradually reduce the resolution as well as increase the number
of feature map channels. ResNet has five architectures: ResNet-18, ResNet-34,
ResNet-50, ResNet-101 and ResNet-152. Our RoadSeg follows the same naming
rule of ResNet. cn, the number of feature map channels (see Fig. 1) varies with
respect to the adopted ResNet architecture. Specifically, c0–c4 are 64, 64, 128,
256 and 512, respectively, for ResNet-18 and ResNet-34, and are 64, 256, 512,
1024 and 2048, respectively, for ResNet-50, ResNet-101 and ResNet-152.f The decoder consists of two different types of modules: a) feature extractors
Fi,j and b) upsampling layers Ui, j, which are connected densely to realize flexible
feature fusion. The feature extractor is employed to extract features from the SNE-RoadSeg 7 fused feature maps, and it ensures that the feature map resolution is unchanged. The upsampling layer is employed to increase the resolution and decrease the
feature map channels. Three convolutional layers in the feature extractor and
the upsampling layer have the same kernel size of 3 × 3, the same stride of 1 and
the same padding of 1. 4 carla.org 4.1
Datasets and Experimental Setup We conduct the experiments of
single-modal CNNs with three setups: a) training with RGB images, b) train-
ing with depth images, and c) training with surface normal images (generated
from depth images using our SNE), which are denoted as RGB, Depth and 4 carla.org R. Fan, H. Wang, P. Cai, and M. Liu 8 (a)
(b)
(c)
(d)
Average angular error (°)
LINE-MOD
SRI
SNE (Ours)
Fig. 3: Qualitative and quantitative results on the DIODE dataset: (a) RGB
images; (b)–(d): the angular error maps obtained using our proposed SNE, SRI
[2] and LINE-MOD [22], respectively. (a)
Average angular error (°) (b)
(c)
(d) (b) (d) (a) (c) Fig. 3: Qualitative and quantitative results on the DIODE dataset: (a) RGB
images; (b)–(d): the angular error maps obtained using our proposed SNE, SRI
[2] and LINE-MOD [22], respectively. SNE-Depth, respectively. Similarly, the experiments of data-fusion CNNs are
conducted using two setups: training using RGB-D vision data, with and without
our SNE embedded, which are denoted as RGBD and SNE-RGBD, respec-
tively. To compare the performances between our proposed RoadSeg and other
state-of-the-art CNNs, we train our RoadSeg with the same setups as for the
data-fusion CNNs on the three datasets. Moreover, we re-train our SNE-RoadSeg
for the result submission to the KITTI road benchmark [15]. The experimen-
tal results are presented in Section 4.3. Additionally, the ablation study of our
SNE-RoadSeg is provided in Section 4.4. SNE-Depth, respectively. Similarly, the experiments of data-fusion CNNs are
conducted using two setups: training using RGB-D vision data, with and without
our SNE embedded, which are denoted as RGBD and SNE-RGBD, respec-
tively. To compare the performances between our proposed RoadSeg and other
state-of-the-art CNNs, we train our RoadSeg with the same setups as for the
data-fusion CNNs on the three datasets. Moreover, we re-train our SNE-RoadSeg
for the result submission to the KITTI road benchmark [15]. The experimen-
tal results are presented in Section 4.3. Additionally, the ablation study of our
SNE-RoadSeg is provided in Section 4.4. Five common metrics are used for the performance evaluation of freespace de-
tection: accuracy, precision, recall, F-score and the intersection over union (IoU). 4.2
Performance Evaluation of Our SNE We simply set gx =
1
Z(x−1,y) −
1
Z(x+1,y) and gy =
1
Z(x,y−1) −
1
Z(x,y+1) to evaluate the
accuracy of our proposed SNE. In addition, we also compare it with two well-
known surface normal estimation approaches: SRI [2] and LINE-MOD [22]. The
qualitative and quantitative comparisons are shown in Fig. 3. It can be observed
that our proposed SNE outperforms SRI and LINE-MOD for both indoor and
outdoor scenarios. 4.1
Datasets and Experimental Setup Their corresponding definitions are as follows: Accuracy =
ntp+ntn
ntp+ntn+nfp+nfn , Preci-
sion =
ntp
ntp+nfp , Recall =
ntp
ntp+nfn , F-score =
2n2
tp
2n2
tp+ntp(nfp+nfn) and IoU =
ntp
ntp+nfp+nfn ,
where ntp, ntn, nfp and nfn represents the true positive, true negative, false pos-
itive, and false negative pixel numbers, respectively. In addition, the stochastic
gradient descent with momentum (SGDM) optimizer is utilized to minimize the
loss function, and the initial learning rate is set to 0.001. Furthermore, we adopt
the early stopping mechanism on the validation subset to avoid over-fitting. The
performance is then quantified using the testing subset. 4.3
Performance Evaluation of Our SNE-RoadSeg In this subsection, we evaluate the performance of our proposed SNE-RoadSeg-
152 (abbreviated as SNE-RoadSeg) both qualitatively and quantitatively. Ex- SNE-RoadSeg 9 Single-modal
Data-fusion
(i)
(ii)
(iii)
(iv)
(v)
(7)
(8)
(9)
(10)
(11)
(1)
(2)
(3)
(4)
(5)
(6)
(a)
Single-modal
Data-fusion
(7)
(8)
(9)
(10)
(11)
(i)
(ii)
(iii)
(iv)
(v)
(1)
(2)
(3)
(4)
(5)
(6)
(b)
Fig. 4: Examples of the experimental results on (a) the SYNTHIA road dataset
and (b) our R2D road dataset: (i) RGB, (ii) Depth, (iii) SNE-Depth (Ours),
(iv) RGBD and (v) SNE-RGBD (Ours); (1) DeepLabv3+ [9], (2) U-Net [26],
(3) SegNet [3], (4) GSCNN [30], (5) DUpsampling [32], (6) DenseASPP [37], (7)
FuseNet [19], (8) RTFNet [29], (9) Depth-aware CNN [35], (10) MFNet [18] and
(11) R
dS
(O
) Th t
iti
f l
ti
d f l
iti
i
l Single-modal
Data-fusion
(i)
(ii)
(iii)
(iv)
(v)
(7)
(8)
(9)
(10)
(11)
(1)
(2)
(3)
(4)
(5)
(6)
(a) (11) Single-modal
Data-fusion
(7)
(8)
(9)
(10)
(11)
(i)
(ii)
(iii)
(iv)
(v)
(1)
(2)
(3)
(4)
(5)
(6)
(b) (b) Fig. 4: Examples of the experimental results on (a) the SYNTHIA road dataset
and (b) our R2D road dataset: (i) RGB, (ii) Depth, (iii) SNE-Depth (Ours),
(iv) RGBD and (v) SNE-RGBD (Ours); (1) DeepLabv3+ [9], (2) U-Net [26],
(3) SegNet [3], (4) GSCNN [30], (5) DUpsampling [32], (6) DenseASPP [37], (7)
FuseNet [19], (8) RTFNet [29], (9) Depth-aware CNN [35], (10) MFNet [18] and
(11) RoadSeg (Ours). The true positive, false negative and false positive pixels
are shown in green, red and blue, respectively. R. Fan, H. Wang, P. Cai, and M. Liu 10 KITTI
SYNTHIA
R2D
5: Performance comparison (%) among DeepLabv3+ [9], U-Net [26], Se
GSCNN [30], DUpsampling [32] and DenseASPP [37] with and withou
E embedded, where
RGB,
Depth, and
SNE-Depth (Ours). KITTI SYNTHIA Fig. 5: Performance comparison (%) among DeepLabv3+ [9], U-Net [26], SegNet
[3], GSCNN [30], DUpsampling [32] and DenseASPP [37] with and without our
SNE embedded, where
RGB,
Depth, and
SNE-Depth (Ours). 11 SNE-RoadSeg KITTI KITTI
SYNTHIA
R2D
nce comparison (%) among FuseNet [19] RTFNet [29] d SYNTHIA
i
(%)
F
N SYNTHIA
ison (%) among FuseN
8] and our RoadSeg wit R2D KITTI
Performance comparis R2D
et [19] RTFNet [29] KITTI
SYNTHIA
R2D
Fig. 4.3
Performance Evaluation of Our SNE-RoadSeg 6: Performance comparison (%) among FuseNet [19], RTFNet [29], depth-
aware CNN [35], MFNet [18] and our RoadSeg with and without our SNE em-
bedded, where
RGBD and
SNE-RGBD (Ours). Fig. 6: Performance comparison (%) among FuseNet [19], RTFNet [29], depth-
aware CNN [35], MFNet [18] and our RoadSeg with and without our SNE em-
bedded, where
RGBD and
SNE-RGBD (Ours). amples of the experimental results on the SYNTHIA road dataset [21] and our
R2D road dataset are shown in Fig. 4. We can clearly observe that the CNNs
with RGB images as inputs suffer greatly from poor illumination conditions. Moreover, the CNNs with our SNE embedded generally perform better than amples of the experimental results on the SYNTHIA road dataset [21] and our
R2D road dataset are shown in Fig. 4. We can clearly observe that the CNNs
with RGB images as inputs suffer greatly from poor illumination conditions. Moreover, the CNNs with our SNE embedded generally perform better than R. Fan, H. Wang, P. Cai, and M. Liu 12 Table 1: The KITTI road benchmark results, where the best results are in bold
type. Please note that we only compare our method with published works. Method
MaxF (%)
AP (%)
PRE (%)
REC (%)
Rank
RBNet [10]
93.21
89.18
92.81
93.60
21
TVFNet [17]
95.34
90.26
95.73
94.94
16
LC-CRF [16]
95.68
88.34
93.62
97.83
13
LidCamNet [5]
96.03
93.93
96.23
95.83
7
RBANet [28]
96.30
89.72
95.14
97.50
6
SNE-RoadSeg (Ours)
96.75
94.07
96.90
96.61
2
(a)
(b)
(c)
(d)
(e)
(f)
Fig. 7: Examples on the KITTI road benchmark, where rows (a)–(f) show the
freespace detection results obtained by RBNet [10], TVFNet [17], LC-CRF [16],
LidCamNet [5], RBANet [28] and our proposed SNE-RoadSeg, respectively. The
true positive, false negative and false positive pixels are shown in green, red and
blue, respectively. Table 1: The KITTI road benchmark results, where the best results are in bold
type. Please note that we only compare our method with published works. Method
MaxF (%)
AP (%)
PRE (%)
REC (%)
Rank
RBNet [10]
93.21
89.18
92.81
93.60
21
TVFNet [17]
95.34
90.26
95.73
94.94
16
LC-CRF [16]
95.68
88.34
93.62
97.83
13
LidCamNet [5]
96.03
93.93
96.23
95.83
7
RBANet [28]
96.30
89.72
95.14
97.50
6
SNE-RoadSeg (Ours)
96.75
94.07
96.90
96.61
2 Table 1: The KITTI road benchmark results, where the best results are in bold
type. 4.3
Performance Evaluation of Our SNE-RoadSeg Please note that we only compare our method with published works. (a)
(b)
(c)
(d)
(e)
(f)
Fig. 7: Examples on the KITTI road benchmark, where rows (a)–(f) show the
freespace detection results obtained by RBNet [10], TVFNet [17], LC-CRF [16],
LidCamNet [5], RBANet [28] and our proposed SNE-RoadSeg, respectively. The
true positive, false negative and false positive pixels are shown in green, red and
blue, respectively. (a)
(b)
(c)
(d)
(e)
(f) (a)
(b)
(c)
(d)
(e)
(f)
Fig. 7: Examples on the KITTI road benchmark, where rows (a)–(f) show the
freespace detection results obtained by RBNet [10], TVFNet [17], LC-CRF [16],
LidCamNet [5], RBANet [28] and our proposed SNE-RoadSeg, respectively. The
true positive, false negative and false positive pixels are shown in green, red and
blue, respectively. Fig. 7: Examples on the KITTI road benchmark, where rows (a)–(f) show the
freespace detection results obtained by RBNet [10], TVFNet [17], LC-CRF [16],
LidCamNet [5], RBANet [28] and our proposed SNE-RoadSeg, respectively. The
true positive, false negative and false positive pixels are shown in green, red and
blue, respectively. they do without our SNE embedded. The corresponding quantitative compar-
isons are given in Fig 5 and Fig. 6. Readers can see that the IoU increases by
approximately 2-12% for single-modal CNNs and by about 1-7% for data-fusion
CNNs, while the F-score increases by around 1-7% for single-modal CNNs and
by about 1-4% for data-fusion CNNs. We demonstrate that our proposed SNE
can make the road areas become highly distinguishable, and thus, it will benefit
all state-of-the-art CNNs for freespace detection. Furthermore, from Fig 5 and Fig. 6, we can observe that RoadSeg itself
outperforms all other CNNs. We demonstrate that the densely-connected skip
connections utilized in our proposed RoadSeg can help achieve flexible feature
fusion and smooth the gradient flow to generate accurate freespace detection
results. Also, RoadSeg with our SNE embedded performs better than all other
CNNs with our SNE embedded. An increase of approximately 1.4-14.7% is wit-
nessed on the IoU, while the F-score increases by about 0.7-8.8%. SNE-RoadSeg 13 Fig. 8: Unsatisfactory results obtained using the KITTI road dataset. The true
positive, false negative and false positive pixels are shown in green, red and blue,
respectively. Fig. 8: Unsatisfactory results obtained using the KITTI road dataset. The true
positive, false negative and false positive pixels are shown in green, red and blue,
respectively. 4.3
Performance Evaluation of Our SNE-RoadSeg In addition, we compare our proposed method with five state-of-the-art CNNs
published on the KITTI road benchmark [15]. Examples of the experimental
results are shown in Fig. 7. The quantitative comparisons are given in Table
1, which shows that our proposed SNE-RoadSeg achieves the highest MaxF
(maximum F-score), AP (average precision) and PRE (precision), while LC-
CRF [16] achieves the best REC (recall). Our freespace detection method is the
second best on the KITTI road benchmark [15]. Fig. 8 presents several unsatisfactory results of our SNE-RoadSeg on the
KITTI road dataset [15]. Since the 3D points on freespace and sidewalks possess
very similar surface normals, our proposed approach can sometimes mistakenly
recognize part of sidewalks as freespace, especially when the textures of the road
and sidewalks are similar. We believe this can be improved by leveraging sur-
face normal gradient features, as there usually exists a clear boundary between
freespace and sidewalks (due to their differences in height). 5
Conclusion The main contributions of this paper include: a) a module named SNE, capable
of inferring surface normal information from depth/disparity images with both
high precision and efficiency; b) a data-fusion CNN architecture named Road-
Seg, capable of fusing both RGB and surface normal information for accurate
freespace detection; and c) a publicly available synthetic dataset for semantic
driving scene segmentation. To demonstrate the feasibility and effectiveness of
the proposed SNE module, we embedded it into ten state-of-the-art CNNs and
evaluated their performances for freespace detection. The experimental results
illustrated that our introduced SNE can benefit all these CNNs for freespace
detection. Furthermore, our proposed data-fusion CNN architecture RoadSeg is
most compatible with our proposed SNE, and it outperforms all other CNNs
when detecting drivable road regions. 4.4
Ablation Study skip connections (SSCs), which employs the skip connections only at the same-
scale feature maps of the encoder and decoder (like U-Net). Table 2 verifies the
superiority of the densely-connected skip connections, which helps to achieve
flexible feature fusion and to smooth the gradient flow to generate accurate
freespace detection results, as analyzed in Section 4.3. skip connections (SSCs), which employs the skip connections only at the same-
scale feature maps of the encoder and decoder (like U-Net). Table 2 verifies the
superiority of the densely-connected skip connections, which helps to achieve
flexible feature fusion and to smooth the gradient flow to generate accurate
freespace detection results, as analyzed in Section 4.3. 4.4
Ablation Study In this subsection, we conduct ablation studies on our R2D road dataset to
validate the effectiveness of the architecture for our RoadSeg. The performances
of different architectures are provided in Table 2.f Firstly, we replace the backbone of RoadSeg with different ResNet architec-
tures. The quantitative results are given in Table 2. The superior performance of
our choice is as expected, because ResNet-152 has also presented the best image
classification performance among the five ResNet architectures [20]. Then, we remove one encoder from RoadSeg to evaluate its performance on
single-modal vision data. We conduct five experiments: a) training with RGB
images, denoted as RGB; b) training with depth images, denoted as Depth;
c) training with depth images, denoted as SNE-Depth; d) training with four-
channel RGB-D vision data, denoted as RGBD-C; and e) training with four-
channel RGB-D vision data, denoted as SNE-RGBD-C. From Table 2, we can
observe that our choice outperforms the single-modal architecture with respect
to different modalities of training data, proving that the data fusion via a two-
encoder architecture can benefit the freespace detection. It should be noted that
although the single-modal architectures cannot provide competitive results, our
proposed SNE still benefits them for better freespace detection performance.f To further validate the effectiveness of our choice, we replace the densely-
connected skip connections in the decoder with two different architectures: a) no
skip connections (NSCs), which totally removes the skip connections; b) sparse R. Fan, H. Wang, P. Cai, and M. Liu 14 Table 2: Performance comparison (%) among different architectures and setups
on our R2D road dataset. The best results are shown in bold font. Architecture
Setup
Accuracy Precision
Recall
F-Score
IoU
RoadSeg-18
SNE-RGBD
93.6
93.5
91.3
92.4
85.9
RoadSeg-34
95.5
96.3
93.0
94.6
89.8
RoadSeg-50
96.8
97.5
95.2
96.3
92.9
RoadSeg-101
98.0
98.2
97.1
97.6
95.4
RoadSeg-152
RGB
94.0
91.9
93.8
92.8
86.6
Depth
96.7
97.6
94.6
96.1
92.4
SNE-Depth
97.6
98.9
95.5
97.2
94.5
RGBD-C
95.1
92.8
95.6
94.2
89.0
SNE-RGBD-
C
97.0
97.5
95.3
96.4
93.0
RoadSeg-152-NSCs SNE-RGBD
97.9
98.6
96.5
97.5
95.2
RoadSeg-152-SSCs
98.2
99.0
96.8
97.9
95.9
RoadSeg-152
(Ours)
SNE-RGBD
98.6
99.1
97.6
98.3
96.7 Table 2: Performance comparison (%) among different architectures and setups
on our R2D road dataset. The best results are shown in bold font. References 1. Alvarez, J.M., Gevers, T., LeCun, Y., Lopez, A.M.: Road scene segmentation
from a single image. In: European Conference on Computer Vision. pp. 376–389. Springer (2012) 2. Badino, H., Huber, D., Park, Y., Kanade, T.: Fast and accurate computation of
surface normals from range images. In: 2011 IEEE International Conference on
Robotics and Automation. pp. 3084–3091. IEEE (2011) (
)
3. Badrinarayanan, V., Kendall, A., Cipolla, R.: Segnet: A deep convolutional
encoder-decoder architecture for image segmentation. IEEE transactions on pat-
tern analysis and machine intelligence 39(12), 2481–2495 (2017) (
)
(
)
4. Cai, P., Wang, S., Sun, Y., Liu, M.: Probabilistic end-to-end vehicle navigation
in complex dynamic environments with multimodal sensor fusion. IEEE Robotics
and Automation Letters 5, 4218–4224 (2020) 5. Caltagirone, L., Bellone, M., Svensson, L., Wahde, M.: Lidar–camera fusion
for road detection using fully convolutional neural networks. Robotics and Au-
tonomous Systems 111, 125–131 (2019) (
)
6. Chen, L.C., Papandreou, G., Kokkinos, I., Murphy, K., Yuille, A.L.: Semantic
image segmentation with deep convolutional nets and fully connected crfs. CoRR
abs/1412.7062 (2014) 7. Chen, L.C., Papandreou, G., Kokkinos, I., Murphy, K., Yuille, A.L.: Deeplab: Se-
mantic image segmentation with deep convolutional nets, atrous convolution, and
fully connected crfs. IEEE transactions on pattern analysis and machine intelli-
gence 40(4), 834–848 (2017)f 8. Chen, L.C., Papandreou, G., Schroff, F., Adam, H.: Rethinking atrous convolution
for semantic image segmentation. arXiv preprint arXiv:1706.05587 (2017)f 9. Chen, L.C., Zhu, Y., Papandreou, G., Schroff, F., Adam, H.: Encoder-decoder with
atrous separable convolution for semantic image segmentation. In: Proceedings of
the European conference on computer vision (ECCV). pp. 801–818 (2018)i (
)
(
)
10. Chen, Z., Chen, Z.: Rbnet: A deep neural network for unified road and road bound-
ary detection. In: International Conference on Neural Information Processing. pp. 677–687. Springer (2017) (
)
11. Dosovitskiy, A., Ros, G., Codevilla, F., Lopez, A., Koltun, V.: CARLA: An open
urban driving simulator. In: Levine, S., Vanhoucke, V., Goldberg, K. (eds.) Pro-
ceedings of the 1st Annual Conference on Robot Learning. Proceedings of Machine
Learning Research, vol. 78, pp. 1–16. PMLR (13–15 Nov 2017) 12. Fan, R., Jiao, J., Pan, J., Huang, H., Shen, S., Liu, M.: Real-time dense stereo
embedded in a UAV for road inspection. In: Proc. IEEE/CVF Conf. Computer
Vision and Pattern Recognition Workshops (CVPRW). pp. 535–543 (2019) 13. Acknowledgements This work was supported by the National Natural Science Foundation of China,
under grant No. U1713211, and the Research Grant Council of Hong Kong SAR
Government, China, under Project No. 11210017, awarded to Prof. Ming Liu. SNE-RoadSeg SNE-RoadSeg 15 References Fan, R., Ozgunalp, U., Hosking, B., Liu, M., Pitas, I.: Pothole detection based on
disparity transformation and road surface modeling. IEEE Transactions on Image
Processing 29, 897–908 (2019)i 14. Fan, R., Wang, H., Xue, B., Huang, H., Wang, Y., Liu, M., Pitas, I.: Three-filters-
to-normal: An accurate and ultrafast surface normal estimator. arXiv preprint
arXiv:2005.08165 (2020), under peer review 15. Fritsch, J., Kuehnl, T., Geiger, A.: A new performance measure and evaluation
benchmark for road detection algorithms. In: International Conference on Intelli-
gent Transportation Systems (ITSC) (2013) (
) (
)
16. Gu, S., Zhang, Y., Tang, J., Yang, J., Kong, H.: Road detection through crf based
lidar-camera fusion. In: 2019 International Conference on Robotics and Automa-
tion (ICRA). pp. 3832–3838. IEEE (2019) R. Fan, H. Wang, P. Cai, and M. Liu 16 17. Gu, S., Zhang, Y., Yang, J., Alvarez, J.M., Kong, H.: Two-view fusion based convo-
lutional neural network for urban road detection. In: 2019 IEEE/RSJ International
Conference on Intelligent Robots and Systems (IROS). pp. 6144–6149. IEEE (2019) 18. Ha, Q., Watanabe, K., Karasawa, T., Ushiku, Y., Harada, T.: Mfnet: Towards real-
time semantic segmentation for autonomous vehicles with multi-spectral scenes. In: 2017 IEEE/RSJ International Conference on Intelligent Robots and Systems
(IROS). pp. 5108–5115. IEEE (2017) 19. Hazirbas, C., Ma, L., Domokos, C., Cremers, D.: Fusenet: Incorporating depth into
semantic segmentation via fusion-based cnn architecture. In: Asian conference on
computer vision. pp. 213–228. Springer (2016) 20. He, K., Zhang, X., Ren, S., Sun, J.: Deep residual learning for image recognition. In:
Proceedings of the IEEE conference on computer vision and pattern recognition. pp. 770–778 (2016) 21. Hernandez-Juarez, D., Schneider, L., Espinosa, A., Vazquez, D., Lopez, A.M.,
Franke, U., Pollefeys, M., Moure, J.C.: Slanted stixels: Representing san francisco’s
steepest streets. In: British Machine Vision Conference (BMVC), 2017 (2017) 22. Hinterstoisser, S., Cagniart, C., Ilic, S., Sturm, P., Navab, N., Fua, P., Lepetit,
V.: Gradient response maps for real-time detection of textureless objects. IEEE
transactions on pattern analysis and machine intelligence 34(5), 876–888 (2011) ( )
(
)
23. Huang, G., Liu, Z., Van Der Maaten, L., Weinberger, K.Q.: Densely connected
convolutional networks. In: Proceedings of the IEEE conference on computer vision
and pattern recognition. pp. 4700–4708 (2017) 24. Long, J., Shelhamer, E., Darrell, T.: Fully convolutional networks for semantic
segmentation. In: Proceedings of the IEEE conference on computer vision and
pattern recognition. pp. 3431–3440 (2015) 25. References Lu, C., van de Molengraft, M.J.G., Dubbelman, G.: Monocular semantic occu-
pancy grid mapping with convolutional variational encoder–decoder networks. IEEE Robotics and Automation Letters 4(2), 445–452 (2019) 26. Ronneberger, O., Fischer, P., Brox, T.: U-net: Convolutional networks for biomedi-
cal image segmentation. In: International Conference on Medical image computing
and computer-assisted intervention. pp. 234–241. Springer (2015) 27. Sless, L., El Shlomo, B., Cohen, G., Oron, S.: Road scene understanding by oc-
cupancy grid learning from sparse radar clusters using semantic segmentation. In:
Proceedings of the IEEE International Conference on Computer Vision Workshops. pp. 0–0 (2019) 28. Sun, J.Y., Kim, S.W., Lee, S.W., Kim, Y.W., Ko, S.J.: Reverse and boundary
attention network for road segmentation. In: Proceedings of the IEEE International
Conference on Computer Vision Workshops. pp. 0–0 (2019) 29. Sun, Y., Zuo, W., Liu, M.: Rtfnet: Rgb-thermal fusion network for semantic seg-
mentation of urban scenes. IEEE Robotics and Automation Letters 4(3), 2576–
2583 (2019) (
)
30. Takikawa, T., Acuna, D., Jampani, V., Fidler, S.: Gated-scnn: Gated shape cnns
for semantic segmentation. In: Proceedings of the IEEE International Conference
on Computer Vision. pp. 5229–5238 (2019) 31. Thoma, J., Paudel, D.P., Chhatkuli, A., Probst, T., Gool, L.V.: Mapping, local-
ization and path planning for image-based navigation using visual features and
map. In: Proceedings of the IEEE Conference on Computer Vision and Pattern
Recognition. pp. 7383–7391 (2019) 32. Tian, Z., He, T., Shen, C., Yan, Y.: Decoders matter for semantic segmentation:
Data-dependent decoding enables flexible feature aggregation. In: Proceedings of SNE-RoadSeg SNE-RoadSeg 17 the IEEE Conference on Computer Vision and Pattern Recognition. pp. 3126–3135
(2019) the IEEE Conference on Computer Vision and Pattern Recognition. pp. 3126–3135
(2019) 33. Vasiljevic, I., Kolkin, N., Zhang, S., Luo, R., Wang, H., Dai, F.Z., Daniele, A.F.,
Mostajabi, M., Basart, S., Walter, M.R., et al.: Diode: A dense indoor and outdoor
depth dataset. arXiv preprint arXiv:1908.00463 (2019) 34. Wang, H., Fan, R., Sun, Y., Liu, M.: Applying surface normal information in driv-
able area and road anomaly detection for ground mobile robots. In: 2020 IEEE/RSJ
International Conference on Intelligent Robots and Systems (IROS) (2020), to be
published 35. Wang, W., Neumann, U.: Depth-aware cnn for rgb-d segmentation. In: Proceedings
of the European Conference on Computer Vision (ECCV). pp. 135–150 (2018) 36. Wedel, A., Badino, H., Rabe, C., Loose, H., Franke, U., Cremers, D.: B-spline mod-
eling of road surfaces with an application to free-space estimation. References IEEE transac-
tions on Intelligent transportation systems 10(4), 572–583 (2009) 37. Yang, M., Yu, K., Zhang, C., Li, Z., Yang, K.: Denseaspp for semantic segmentation
in street scenes. In: Proceedings of the IEEE Conference on Computer Vision and
Pattern Recognition. pp. 3684–3692 (2018)
|
https://openalex.org/W2302676570
|
http://hrcak.srce.hr/file/216586
|
English
| null |
Familiarisation Aboard Ships of Croatian and Montenegrin Officers
|
Transactions on maritime science
| 2,015
|
cc-by
| 3,737
|
1. INTRODUCTION New officers joining the ship must be aware of their
obligations regarding information of security and safety on
board. Furthermore, familiarisation includes technical skill related
to ships’ equipment. Because of the lack of time during handover
due to a short time of cargo operations in port, handover is
often inadequate. Familiarisation should last more than a month. Officers’ contracts are usually shorter than familiarisation process. Familiarisation time and lack of familiarisation are considered as
period of great risk of human error. According to the research conducted by the British Marine
Accident Investigation Board (MAIB), Canadian Transportation
Safety Board (TSB), and Australian Transportation Safety Board
(ATSB), which carefully studied 350 cases of maritime accidents,
82 to 85 % of all accidents were either directly initiated by
human error or were associated with human error by means
of inappropriate human responding to threat situations. The
incidence of human error in maritime shipping industry does not
arise exclusively from the operator’s failure. There is a wide range
of factors contributing to the occurrence of human error. One of
them is modern technology. Although all equipment aboard has
to be standardized according to the Safety of Life at Sea-SOLAS
Convention requirements, it is considered that inadequately
designed equipment made difficulties to operators and caused
1/3 of major sea accidents. Another aspect affecting the human
error is the interaction between the technology and human
beings i.e. lack of training and familiarisation with equipment’s
operation procedures. A study performed by R. Ziarati and M. Ziarati reveals that inadequate use of navigation equipment
causes 28 % of accidents (Baker et al., 2005). In this case,
“inadequate” refers to the use of equipment in an inappropriate
manner due to insufficient training or non-adapted equipment. In this paper the authors research familiarisation and
handover as safety aspects. Familiarisation Aboard Ships
of Croatian and Montenegrin
Officers Pero Vidana, Tatijana Dlabačb, Goran Jerkovića KEY WORDS Consequently, this leads to a situation where
the operator is forced to handle the devices he/she is not entirely
familiarised with. The operator improvises, relies on the previous
experience and transfers it from the previous devices to the new
ones that do not necessarily have the same mode of operation. These situations open up huge potential for misinterpreting the
information that is provided by automated systems (Foord et al.,
2006). According to British psychologist Lissiane Banbridge, the
core of the problem is that equipment manufacturers and their
designers often tend to displace a human being out of the system
wherever possible but keep on requiring an operator to carry
out the tasks for which they have not found adequate solution. One of the problems that may arise in automated systems is
undoubtedly the poorly designed technology that does not
allow an adequate interaction between the human being and the
equipment. If the interaction between a device and an operator
is not ergonomic, it is likely that the operator will be overloaded
and confused by the information provided by the device. For
example, in case when a number of alarms are given at the same
time, it is possible that critical warnings remain unnoticed (Vidan
et al., 2015). Figure 1. Primary causes of failures in modern exploitation systems. 2. FAMILIARISATION AND HANDOVER ABOARD Modern integrated systems of shipping management have
considerably contributed to the increased safety of sea traffic. Yet, it is necessary to introduce changes within these systems in
order to reduce the share of human error in causing accidents. For
instance, SOLAS regulations define the layout of the bridge, but
do not define its precise design. This allows substantial room for
the manufacturers who, apart from meeting SOLAS requirements,
frequently install additional options to the equipment in order
to boost their price and competitiveness on the market. In this
way, seafarers are moved to a less favourable position, as the
design of technologies forming a modern operating system has
not been developed on the basis of the seafarers’ needs and is
likely to lead an operator to making a mistake. In order to use the
available technology in a fast and efficient way, the operator has
to be thoroughly familiar with all the equipment specification
and modes of use and operation. KEY WORDS ~
~ Familiarisation ~
~ Handover ~
~ Croatian Officers ~
~ Montenegrin Officers Most of regulations and standards applying to ships do not
address complex issues such as cognitive activities associated
with implementing new technologies on board ships. The
number of standards and regulations that exclusively deal with
the relationship between on-board automation and human
beings are relatively low. Almost 80 % of the regulations in a. University of Split, Faculty of Maritime Studies, Split, Croatia TRANSACTIONS ON MARITIME SCIENCE 113 Trans. marit. sci. 2015; 02: 113-118 maritime shipping industry refer to technical rules, whereas the
remaining 20 % refer to humans (Baker et al., 2005). a large number of seafarers employed by forward-thinking
maritime companies attend various forms of training in order to
acquire certificates of competence in operating specific systems,
this does not guarantee that a seafarer will perform his/her duties
using the very device and the very model that he/she familiarised
with during training. For instance, SOLAS Chapter V, regulation 15, lays out the
principles relating to bridge design and design of navigational
systems and equipment. Operators have to cope with the
equipment produced by various manufacturers competing
in additional abilities not obligatory according to SOLAS
requirements and may be used very rarely, bur can lead to
confusion making the overall bridge system inconsistent. The
equipment installed on ships of the same type and purpose may
greatly vary from vessel to vessel, so that operators have to adjust
to a new equipment layout, differing specifications etc., which
represents a potential source of failure. The American National
Transport Safety Board states that almost one out of three failures
is caused by inadequate equipment design, where the lack of
standardisation is the key contributing factor (Albayrak et al.,
2010). The problem of insufficient use of the equipment still
exists. Some of the primary causes of failures resulting in poor
exploitation of modern systems are shown in Figure 1. One of
the essential aspects of this issue is the fact that the equipment
often differs considerably from vessel to vessel, and seafarers
rarely serve the same or similar vessels throughout their career. The end result is the superficial familiarisation with the system. Although every device has a user’s manual offering certain
information and instructions to the operator, the latter is often
unable to read the manual thoroughly upon joining the ship due
to the lack of time. Pero Vidan et al.: Familiarisation Aboard Ships of Croatian and Montenegrin Officers KEY WORDS To achieve this, the operator
has to complete adequate training and familiarisation which will
help him/her to identify all advantages and drawbacks of the
equipment. Proficiency and familiarisation are the key factors in
the prevention of failures and safety hazards. However, although 2. FAMILIARISATION AND HANDOVER ABOARD The new officer crewmembers joining the ship must be
aware of their obligations regarding the use of equipment,
security and safety on board. The main purpose of familiarisation
is to introduce new officers with important safety procedures
on board, emergencies and proper use of ship technology. The
captain shall appoint a qualified person responsible for training
new members of the crew. Although all members of the crew
joining a ship must pass a standardized training by Standards
of Training, Certification and Watch-keeping for Seafarers-STCW, doi: 10.7225/toms.v04.n02.002 114 5. RESULTS Shipboard audits and port-state control results show this
to be a weak area. In many cases, the crew is unfamiliar with the
ship-specific and pollution-prevention equipment (Navigational
Claims Brochure, 2014). Shipboard audits and port-state control results show this
to be a weak area. In many cases, the crew is unfamiliar with the
ship-specific and pollution-prevention equipment (Navigational
Claims Brochure, 2014). 200 officers were surveyed, 113 Croatian and 87
Montenegrins. From the total number of 200 officers, 65 (32.5 %)
of them are 25-30 year old, 79 (39.5 %) are 30-40 years old, 33
(16.5 %) are in the range between 40 and 50 years of age and only
23 (11.5 %) are older than 50. Officers are experienced, with less
than 4 years of navigation (7 %), 4-10 years (29 %) and more than
15 years of navigation (29 %) (Figure 2). Handover is a process during the exchange of duties of two
crew members of the same rank. It is usually obtained in ports. It
is a normal procedure on board ships that the new crew member
will be given handover of his or her duties by the person who
is being relieved. During handover the new crew member has
been introduced with the ship, her equipment, specific details of
technology of ship and also duties. During handover, information
about vessel documentation, accounts, logs etc. are usually
exchanged (Shahrokh, 2015; Squire, 2015). Figure 2. Experience of navigation (n=200). 47 (26 %) officers graduated from a maritime faculty and
133 (74 %) finished secondary maritime school (Figure 3). 4. METHODOLOGY The familiarization training will include: The familiarization training will include: Duties to eeks after joining the ship (Unknown, 20 •
Duties to eeks after joining the ship (Unknown, 2015) 4. METHODOLOGY there are specific instructions and requirements specific to each
ship: A research has been performed by the Faculty of Maritime
Studies in Split and Maritime Faculty in Kotor with Croatian and
Montenegrin officers using questionnaires. The subjects of the
questionnaire were the quality of familiarisation and handover in
their companies. The questionnaires were anonymous, but with
the telephone number of the surveyed to enable checking. 20 %
of the questionnaires were checked. •
Instructions on SOLAS and International Convention for the
Prevention of Pollution from Ships-MARPOL Prevention of Pollution from Ships-MARPOL •
Important features of muster list •
Guidelines on how to tackle all kinds of emergency
situations on ships •
Important documents and publications of ships •
Ship’s Fire Control Planfii Questionnaires were written in the Croatian and
Montenegrin languages. Croatia and Montenegro have similar
systems of education of seafaring officers. After the secondary
maritime school seafarers become cadets or engine apprentices. After a year of navigation as cadets, they sit for an exam in the
Harbour Master’s Office for officer in charge on vessels of 500 GT
or more (STCW II/1) or engineer officer in charge on vessels with
propulsion of 750 kW or more (STCW III/1). Captains of vessel
of 3,000 GT or more (STCW II/2) and chief engineers on ships
with propulsion of 3,000 kW and more (STCW III/2) have to have
higher education (maritime faculty) degree and at least one year
of experience as chief mate (deck) or 2nd engineer (engine), or 3
years of experience as deck officer (deck) or 3 years’ experience as
engineer officers (engine). After that they have to pass an exam
in the Harbour Master’s Office. •
Different types of fire-fighting appliances and procedures
to use them During familiarization the officer will be introduced with
specific details of his job depending of the type of ship and
organization of crew on board. This familiarisation also includes
reading manuals of new engines and equipment in order to be
able to properly use them. Except in the performance of watch-keeping at sea and
in port, the new crew members will be informed of all standing
orders of the chief engineer or master. Training for lifesaving appliances and fire appliances should
be completed as soon as possible and no later than two weeks
after the receipt of the ship or the provisions of a single company
ship owner (Unknown, 2012). 3. HYPOTHESIS Handover is usually short and inadequate because of the
lack of time in ports. It depends on the educational background
of the officer, with the same or similar vessel and/or equipment. Handover is usually short and inadequate because of the
lack of time in ports. It depends on the educational background
of the officer, with the same or similar vessel and/or equipment. Familiarisation is a process which starts after embarkation
and lasts for 30-45 days. Because of hi-tech ships it can exceed
45 days. Familiarisation is performed usually according to the
International Safety Management Code-ISM between the new
crew member and a crew member designated by a senior officer. It depends on education of the officer. Familiarisation is a process which starts after embarkation
and lasts for 30-45 days. Because of hi-tech ships it can exceed
45 days. Familiarisation is performed usually according to the
International Safety Management Code-ISM between the new
crew member and a crew member designated by a senior officer. It depends on education of the officer. 47 (26 %) officers graduated from a maritime faculty and
133 (74 %) finished secondary maritime school (Figure 3). TRANSACTIONS ON MARITIME SCIENCE Trans. marit. sci. 2015; 02: 113-118 115 From the total number of 200, 102 officers considered
handover as sufficient and 88 of the officers surveyed considered
familiarisation as sufficient. Figure 3. Education of officers (n=200). doi: 10.7225/toms.v04.n02.002
Pero Vidan et al.: Familiarisation Aboard Ships of Croatian and Montenegrin Officers
Figure 3. Education of officers (n=200). Figure 6. Answers of officers who graduated from maritime faculty
(n=47). Figure 7. Answers of officers who finished maritime secondary
school (n=133). Figure 4. Duration of handover (n=200). Figure 5. Duration of familiarisation (n=200). Figure 8. Answers about similarity of vessel where officers sailing. To the question about duration of handover, 64 (31 %)
officers answered “up to 2 hours”. Duration of 2-4 hours was
chosen by 28 officers (14 %), 4-6 hours by 26 officers (13 %). More
than 6 hours was the choice of 84 officers (42 %) (Figure 4). The duration of familiarisation up to 7 days was stated
by 174 officers (87 %), 20 officers (10 %) had 7-15 days of
familiarization. 4 (2 %) officers had 15-30 days’ familiarisation. 2
officer (2 %) replied about 30-45 days’ familiarization (Figure 5)fifi
y
familiarisation as sufficient. 3. HYPOTHESIS From the total number of 133 officers who finished
maritime faculty, 28 considered handover as insufficient and 33
thought the same about familiarisation (Figure 6). The officers
who finished only maritime secondary school thought that
handover was insufficient were 63 and 20 thought the same
about familiarisation (Figure 7). 58 % of officers sail on board Figure 3. Education of officers (n=200). Figure 6. Answers of officers who graduated from maritime faculty
(n=47). Figure 6. Answers of officers who graduated from maritime faculty
(n=47). To the question about duration of handover, 64 (31 %)
officers answered “up to 2 hours”. Duration of 2-4 hours was
chosen by 28 officers (14 %), 4-6 hours by 26 officers (13 %). More
than 6 hours was the choice of 84 officers (42 %) (Figure 4). Figure 4. Duration of handover (n=200). Figure 7. Answers of officers who finished maritime secondary
school (n=133). Figure 4. Duration of handover (n=200). From the total number of 133 officers who finished
maritime faculty, 28 considered handover as insufficient and 33
thought the same about familiarisation (Figure 6). The officers
who finished only maritime secondary school thought that
handover was insufficient were 63 and 20 thought the same
about familiarisation (Figure 7). 58 % of officers sail on board The duration of familiarisation up to 7 days was stated
by 174 officers (87 %), 20 officers (10 %) had 7-15 days of
familiarization. 4 (2 %) officers had 15-30 days’ familiarisation. 2 fi
officer (2 %) replied about 30-45 days’ familiarization (Figure 5) Figure 5. Duration of familiarisation (n=200). doi: 10.7225/toms.v04.n02.002
Pero Vidan et al.: Familiarisation Aboard Ships of Croatian and Montenegrin Officers
Figure 5. Duration of familiarisation (n=200). Figure 8. Answers about similarity of vessel where officers sailing. ( g
)fi Figure 8. Answers about similarity of vessel where officers sailing. Figure 8. Answers about similarity of vessel where officers sailing. Pero Vidan et al.: Familiarisation Aboard Ships of Croatian and Montenegrin Officers 116 doi: 10.7225/toms.v04.n02.002 sister ships. The remaining 42 % do not sail on board sister ships,
but on ships using the same technology. sister ships. The remaining 42 % do not sail on board sister ships,
but on ships using the same technology. Shippers, in order to reduce familiarisation time and
handover, usually embark seafarers, especially officers on sister
ships or ships of the same technology. REFERENCES Albayrak, T. and Ziarati, R., (2010), Training: Onboard and Simulation Based
Familiarisation and Skill Enhancement to Improve the Performance of Seagoing
Crew, Proc. International Conference on Human Performance at Sea HPAS 2010,
Glasgow, Scotland, UK,June 16 – 18, pp. 11 – 21., available at: http://www.egmdss. com/pictures/press/2010_06_16_tr_HPAS2010_improve_the_performance_of_
seagoing_crew.pdf Baker, C.C. and McCafferty, D.B., (2005), Accident database review of human
element concerns: what do the results mean for classification?, Proc. International
Conference on Human Factors in Ship Design, Safety and Operation, London, UK,
February 23 - 24, availabe at: http://citeseerx.ist.psu.edu/viewdoc/download?doi=1
0.1.1.437.6892&rep=rep1&type=pdf Although 42 % of surveyed officers had handover longer
than 6 hours, 59.58 % of officers considered this period as
insufficient. Taking into consideration Figure 8, it is to conclude
that handover requires more time. The interview with the officers
also discovered that, in an inversely proportional manner the
ship administration has recently increased while handover has
become shorter. Bielic,,T. and Zec, D., (2004), Influence of Ship Technology and Work Organisation on
Fatigue, Pomorski zbornik, 42 (1), pp. 263-276. Foord, A. G. and Gulland, W. G., (2006), Can technology eliminate human error?,
available
at:
http://wildeanalysis.co.uk/system/downloads/371/original/Can_
Technology_Eliminate_Human_Error.pdf, [accessed 21 April 2015.]. Foord, A. G. and Gulland, W. G., (2006), Can technology eliminate human error?,
available
at:
http://wildeanalysis.co.uk/system/downloads/371/original/Can_
Technology_Eliminate_Human_Error.pdf, [accessed 21 April 2015.]. 6. DISCUSSION This survey includes 200 seafarers, officers from Croatia
and Montenegro. There was no previous research about
familiarisation and handover, so this statistics does not include
control group and deviation data. 200 questionnaires are not a representative number, but
the research shows significant phenomena. There has not been a
similar research up to date, so that the amount of deviation from
the control number could not be calculated. The authors expect
this survey to lead to a future research about familiarisation of
crew aboard ships. In a future research it is necessary to include data about
experience and how experience affects familiarisation and
handover. For better results, officers should not navigate on sister
ships. It is also recommended to include shippers and agencies
that embark seafarers and try to research their statements and
opinions. Most of the officers surveyed finished a secondary maritime
school - 74 % (Figure 5) and most of the surveyed have experience
of sailing 10-15 years (Figure 4). Comparing Figure 6, which
shows answers of officers graduated from a maritime faculty, and
Figure 7, which shows answers provided by those having finished
a secondary maritime school, it’s obvious that handover and
familiarisation do not depend on the education level. In a future
survey it is recommended to compare how experience in sailing
and training affects familiarisation and handover. During this survey, a researcher found out that in a future
survey it would be very important to analyse how officer rank
and the size of ship and engine affect the time of handover and
familiarisation. For the representativeness of the number of surveyed, it
is necessary to extend the survey to other nations with different
systems of education and promotion and increase the number of
surveyed to at least 1,000. Results are the same for Croatian and Montenegrin officers,
so there was no need for a detailed analysis by nationality. Results are the same for Croatian and Montenegrin officers,
so there was no need for a detailed analysis by nationality. 42 % of the surveyed answered that they had handover
longer than 42 % which s consider ad enough (Figure 4). 87 %
of surveyed had familiarisation up to 7 days which consider as
insufficient. Most officers (Figure 6) are satisfied with time of
familiarisation, although it was up to 7 days (Figure 4). 6. DISCUSSION This result
is confused but it is consequence of fact that 58 % of officers
have sister ship as last ship of sailing (Figure 8), and 98 % officers
has last ship with same technology as previous. fi 42 % of the surveyed answered that they had handover
longer than 42 % which s consider ad enough (Figure 4). 87 %
of surveyed had familiarisation up to 7 days which consider as
insufficient. Most officers (Figure 6) are satisfied with time of
familiarisation, although it was up to 7 days (Figure 4). This result
is confused but it is consequence of fact that 58 % of officers
have sister ship as last ship of sailing (Figure 8), and 98 % officers
has last ship with same technology as previous. Vidan, P., Bukljaš Skočibušić, M. and Rubić, J., (2015), Influence of New Technology
on Human Error, Proc. 17th International Conference on Transport Science, Portorož,
Slovenia, May 21 – 22, pp. 554-562. Pero Vidan et al.: Familiarisation Aboard Ships of Croatian and Montenegrin Officers Unknown, (2013), Familiarization for New Crew Members on Board of the Ship,
available at: http://sea-jobs.net/encyclopediaen/190, [accessed 1 September 2015.]. Unknown, (2013), Familiarization for New Crew Members on Board of the Ship,
available at: http://sea-jobs.net/encyclopediaen/190, [accessed 1 September 2015.].
Unknown, (2012), Procedure for Ship Familiarization for New Crew Members on
Ships, available at: http://www.marineinsight.com/misc/marine-safety/procedure-
for-ship-familiarization-for-new-crew-members-on-ships/, [accessed 30 august
2015.]. Unknown, (2012), Procedure for Ship Familiarization for New Crew Members on
Ships, available at: http://www.marineinsight.com/misc/marine-safety/procedure-
for-ship-familiarization-for-new-crew-members-on-ships/, [accessed 30 august
2015.]. doi: 10.7225/toms.v04.n02.002 7. CONCLUSION Navigational Claims Brochure, (2014), available at: http://www.swedishclub.com/
upload/Publications/navigational-claims-brochure.pdf, [accessed 11 February
2015.]. Navigational Claims Brochure, (2014), available at: http://www.swedishclub.com/
upload/Publications/navigational-claims-brochure.pdf, [accessed 11 February
2015.]. Familiarisation and handover are considered as very
important for the safety and security of ship. Technological
innovations prolong them. Seafarers+ contracts become
shorter. In some cases, e.g. on offshore vessels, where contracts
have duration of one month aboard and one month at home,
familiarisation can last as long as the contract period. Shahrokh, K., (2015???), Familiarisation on Board, available at: http://www.he-alert. org/filemanager/...-/he01095.pdf, [accessed 12 June 2015.]. Squire, D., (2006), The Human Element in Shipping, available at: http://www. he-alert.org/filemanager/root/site_assets/standalone_pdfs_0355-/he00350.pdf,
[accessed 1 September 2015.]. STCW and International Maritime Organisation-IMO are
introduced with the problem of familiarisation and handover. Because of this, they prescribed familiarisation trainings for
special ships and equipment. Unknown, (2006), Guidance to Masters, 2nd Edition, Arendal, Norway: Gard,
available at: http://www.gard.no/Content/20651969/GTM_web.pdf, [accessed 1
September 2015.]. Trans. marit. sci. 2015; 02: 113-118 TRANSACTIONS ON MARITIME SCIENCE 117 118 doi: 10.7225/toms.v04.n02.002
|
https://openalex.org/W4226332163
|
https://research.gold.ac.uk/id/eprint/29928/7/141b52fa-a47a-4699-8d28-e51d9e5306d8_15588_-_kevin_malone_v3.pdf
|
English
| null |
Bringing Lived Lives to Swift’s Asylum: a psychiatric hospital perspective
|
Wellcome open research
| 2,022
|
cc-by
| 17,101
|
Bringing Lived Lives to Swift’s Asylum: a psychiatric
hospital perspective [version 3; peer review: 2 approved]
Kevin M. Malone
1, Eimear Cleary1, Cecily C. Kelleher1, Janis Jefferies2,
Abbie Lane
1, James V. Lucey3, Seamus McGuiness4 3Department of Psychiatry, St. Patrick's University Hospital, Dublin, Co. Dublin, Ireland
45GMIT Centre for Creative Arts & Media, Galway Mayo Institute of Technology, Galway, Ireland Open Peer Review
Approval Status
1
2
version 3
(revision)
29 Mar 2022
view
version 2
(revision)
20 Dec 2021
view
view
version 1
19 Apr 2021
view First published: 19 Apr 2021, 6:85
https://doi.org/10.12688/wellcomeopenres.15588.1
Second version: 20 Dec 2021, 6:85
https://doi.org/10.12688/wellcomeopenres.15588.2
Latest published: 29 Mar 2022, 6:85
https://doi.org/10.12688/wellcomeopenres.15588.3
v3 First published: 19 Apr 2021, 6:85
https://doi.org/10.12688/wellcomeopenres.15588.1
Second version: 20 Dec 2021, 6:85
https://doi.org/10.12688/wellcomeopenres.15588.2
Latest published: 29 Mar 2022, 6:85
https://doi.org/10.12688/wellcomeopenres.15588.3
v3 First published: 19 Apr 2021, 6:85
https://doi.org/10.12688/wellcomeopenres.15588.1
Second version: 20 Dec 2021, 6:85
https://doi.org/10.12688/wellcomeopenres.15588.2
Latest published: 29 Mar 2022, 6:85
https://doi.org/10.12688/wellcomeopenres.15588.3
v3 view
view Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 RESEARCH ARTICLE Keywords
Stigma, Mental Illness, Psychiatric Hospitalization, Suicidal Ideation,
Psychoeducation Keywords
Stigma, Mental Illness, Psychiatric Hospitalization, Suicidal Ideation,
Psychoeducation Corresponding author: Kevin M. Malone (k.malone@st-vincents.ie)
Author roles: Malone KM: Conceptualization, Data Curation, Formal Analysis, Funding Acquisition, Investigation, Methodology, Project
Administration, Supervision, Writing – Original Draft Preparation, Writing – Review & Editing; Cleary E: Formal Analysis, Project
Administration, Writing – Original Draft Preparation, Writing – Review & Editing; Kelleher CC: Conceptualization, Methodology, Writing –
Original Draft Preparation, Writing – Review & Editing; Jefferies J: Conceptualization, Methodology, Writing – Original Draft Preparation;
Lane A: Methodology, Writing – Original Draft Preparation, Writing – Review & Editing; Lucey JV: Funding Acquisition, Investigation,
Writing – Review & Editing; McGuiness S: Conceptualization, Data Curation, Formal Analysis, Funding Acquisition, Investigation,
Methodology, Project Administration, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing
Competing interests: No competing interests were disclosed. Grant information: This work was supported by the Wellcome Trust [208947]. This work was also supported by a European Union
Research and Innovation Science in Society grant. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Copyright: © 2022 Malone KM et al. This is an open access article distributed under the terms of the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. How to cite this article: Malone KM, Cleary E, Kelleher CC et al. Bringing Lived Lives to Swift’s Asylum: a psychiatric hospital
perspective [version 3; peer review: 2 approved] Wellcome Open Research 2022, 6:85
https://doi.org/10.12688/wellcomeopenres.15588.3
First published: 19 Apr 2021, 6:85 https://doi.org/10.12688/wellcomeopenres.15588.1 Abstract Background: Few “interventions” around suicide and stigma have
reached into psychiatric institutions. Lived Lives is a science-arts
approach to addressing suicide and stigma, informed by a
psychobiographical and visual arts autopsy. The resulting artworks
and mediated exhibition ( Lived Lives), has facilitated dialogue,
response and public action around stigma-reduction, consistent with a
community intervention. Recent evidence from Lived Lives moved us to
consider how it may situate within a psychiatric hospital. Methods: Lived Lives manifested in St. Patrick’s University Hospital
(Ireland’s oldest and largest psychiatric hospital) in November 2017. A mixed-methods approach was used to evaluate the exhibition as a
potential intervention to address stigma around suicide, with
quantitative and qualitative data collected via written questionnaire
and oral data collected via video documentation. Bereavement
support was available. A Clinician and an artist also provided
independent evaluation. Leah Quinlivan
, The University of
Manchester, Manchester Academic Health
Science Centre, Manchester, UK
University of Manchester, Manchester, UK
1. Rahul Shidhaye
, Pravara Institute of
Medical Sciences, Pune, India
2. Any reports and responses or comments on the
article can be found at the end of the article. Results: 86 participants engaged with the exhibition, with 68
completing questionnaire data. Audiences included service users,
policy makers, health professionals, senior hospital administrators
and members of the public. 62% of participants who completed
questionnaires were suicide-bereaved; 46% had experienced a mental
health difficulty, and 35% had been suicidal in the past. 91% thought
Lived Lives could be of benefit in the aftermath of a suicide death. Half
of participants thought Lived Lives could help reduce suicidal feelings,
whereas 88% thought it could benefit those with Mental Health
difficulties. The emotional response was of a visceral nature, including
fear, anger, sadness, disgust and anxiety. Conclusions: Lived Lives sits comfortably in discomfort, unafraid to call
out the home-truths about stigma and its pervasive and pernicious Page 1 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 impact, and with restoring identity at its core. Lived Lives can operate
within a psychiatric hospital, as well as in community. The challenge is
to move it forward for greater exposure and impacts in at-risk
communities. Introduction and context
Depression and suicide in Ireland Depression and suicide in Ireland p
Over the past decade (2005 – 2016), there has been an aver-
age of 590 suicide and open verdict deaths per year in Ireland
(504 suicide deaths, 86 open verdict according to the CSO)
(Central Statistics Office, 2019). International research estimates
that between 30% and 60% of suicide deaths suffer from a
major depressive disorder (MDD) (Conwell et al., 1996; Foster
et al., 1997; Harwood et al., 2001; Henriksson et al., 1993). Assuming a 60% suicide rate (Hasin et al., 2018), in excess
of 300 suicide deaths in Ireland annually suffer from MDD. (approximately 60 females, 242 males). It has been estimated that
every suicide death impacts 71 members of society (Mallon &
Galway, 2015). Therefore, each year, in excess of 20,000
living individuals in Ireland are impacted by an MDD-related
suicide death. This impact (or toll) is cumulative, such that
over a decade, in excess of 200,000 living individuals in
Ireland carry this loss. On the other hand, the vast majority of
those suffering from MDD each year do not die by suicide. For every one suicide death each year with MDD in Ireland,
there are anywhere between 800 and 1,100 people living with
MDD (based on a 12 month prevalence of MDD in 18–64
year olds of 8–12% – 65% female, 35% male). Effective
in-patient and out-patient treatment for MDD significantly
lowers the risk of suicide (Brodsky et al., 2018; Möller &
Henkel, 2005). However, between 40% and 60% of those who
die by suicide do so at the 1st attempt (Bostwick et al., 2016),
and therefore never benefit from all the post suicide attempt
intervention (in-patient or out-patient) and treatment protocols
(and statutory and voluntary investments). Stigma of mental
illness negatively impacts recovery (Corrigan & Miller, 2004;
Henderson et al., 2014). There is very little research on projects
which challenge the stigma around mental illness and suicide
(Carpiniello & Pinna, 2017; Rüsch et al., 2005). Lived Lives project. There are currently very few “interventions”
around suicide and stigma with evaluation as an integral compo-
nent, and fewer still that have reached into psychiatric institutions. According to the Wellcome Trust, Science - Arts collaborations
are “especially effective for complex health / society challenges
to generate new knowledge and understanding” (Malone et al.,
2017). Lived Lives is a mixed methods science-arts approach to
addressing suicide and stigma. Corresponding author: Kevin M. Malone (k.malone@st-vincents.ie) Author roles: Malone KM: Conceptualization, Data Curation, Formal Analysis, Funding Acquisition, Investigation, Methodology, Project
Administration, Supervision, Writing – Original Draft Preparation, Writing – Review & Editing; Cleary E: Formal Analysis, Project
Administration, Writing – Original Draft Preparation, Writing – Review & Editing; Kelleher CC: Conceptualization, Methodology, Writing –
Original Draft Preparation, Writing – Review & Editing; Jefferies J: Conceptualization, Methodology, Writing – Original Draft Preparation;
Lane A: Methodology, Writing – Original Draft Preparation, Writing – Review & Editing; Lucey JV: Funding Acquisition, Investigation,
Writing – Review & Editing; McGuiness S: Conceptualization, Data Curation, Formal Analysis, Funding Acquisition, Investigation,
Methodology, Project Administration, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing
Competing interests: No competing interests were disclosed. Page 2 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 illness, suicidal tendencies and who have been psychiatrically
hospitalized are “guilty” by association, and are euphe-
mistically referred to as experiencing “courtesy stigma”
(Phelan & Bromet, 1998). Families who have lost a loved one to
suicide from such illness also experience stigma and its attendant
silence, secrecy and shame (Corrigan & Miller, 2004). For those
patients who have been under in-patient care, such is the level of
stigma that patients can experience being socially ostracised for
years after a single hospital admission, thus bearing the stigma
of mental illness and the stigma of having been institutional-
ised for their illness (Link et al., 1987). The immediate after-
math of psychiatric hospital discharge can be especially
challenging for patients coming to terms with their illness,
and it constitutes a significant period of risk for suicide (Mann
et al., 2005), where stigma and fears of relapse are fre-
quently factors, sharply contrasting with the care and attention
(and investment) afforded those recently discharged from
medical / surgical care (Moses, 2014). Whilst much-needed
research is on-going related to understanding and treating major
psychiatric disorders, there is much less focus on processes
associated with stigma and its impacts in terms of how it is still
endemic in (and following) institutional care, and how it may
undermine recovery (Henderson et al., 2014). Amendments from Version 2
REVISED We have updated this version of the paper to better reflect
research objectives, evaluation methodology and limitations
of the study in terms of measuring longer term impacts and
engaging wider audiences. Any further responses from the reviewers can be found at
the end of the article Introduction and context
Depression and suicide in Ireland A new art-work
– 21g Making Stigma Visible in SPUH involved participants
placing their inked finger on one of the Lived Lives shirt
fragments as a tactile and tangible metaphor for stigma (Figure 2. “21g -Making Stigma Visible” installed in Group Therapy
Room, St. Patrick’s University: a) Lived Lives participant
marking a shirt fragment upon 21g; b) 21g shirt fragments
marked with ink as metaphor for stigma) Paper I for images, Cleary et al., 2021). A new art-work
– 21g Making Stigma Visible in SPUH involved participants
placing their inked finger on one of the Lived Lives shirt
fragments as a tactile and tangible metaphor for stigma (Figure 2. “21g -Making Stigma Visible” installed in Group Therapy
Room, St. Patrick’s University: a) Lived Lives participant
marking a shirt fragment upon 21g; b) 21g shirt fragments
marked with ink as metaphor for stigma) Cleary et al., 2021 https://doi.org/10.12688/wellcomeopenres. 15613.1). Findings from this project indicated that Lived Lives
creates a “safe-space” for catharsis, dialogue, disclosure and
discussion around feelings, and where there is no guilt or secrecy
about experiencing or disclosing past suicidal thoughts or
acts. With this in mind, we were moved to consider how Lived
Lives may respond and be responded to within the context of
manifesting in a psychiatric institution (St. Patrick’s University
Hospital (SPUH), where stigma is chronically apparent both
within and without (Lived Lives Paper II). A brief descrip-
tion of Lived Lives artworks follows. For a more comprehensive
description of artworks, see Lived Lives Paper I (Cleary et al.,
2021
https://doi.org/10.12688/wellcomeopenres.15613.1)
and
Lived Lives: From Tory Island to Swift’s Asylum E-publication
(McGuinness & Malone, 2019: https://bit.ly/35hUjU6) Archive Rooms consists of artworks constructed by the artist
with clothing, writing and other personal objects belonging
to the deceased, donated by the participating families, which
were originally exhibited as a series of rooms with each loved
lost one being represented by their individual belongings. It was
installed in the Medical Library for this event (Figure 3. Archive Rooms, installed in the Medical Library, St. Patrick’s
University Hospital). Lived Lives: Art Installations in St. Patrick’s University
Hospital. 21 g (Figure 1. 21g installed in Group Therapy Room,
St. Patrick’s University Hospital). McGuinness originally
created 21g in 2003 as a visual representation of young male
suicide in Ireland in that year. Introduction and context
Depression and suicide in Ireland It has been informed by a com-
bined psychobiographical autopsy and visual arts autopsy for
studying suicide and its aftermath. The Suicide in Ireland Survey
(2003–2008) documented the lives of 104 suicide victims from
around Ireland as narrated by their families (Malone, 2013),
and “Lived Lives”, a parallel visual arts autopsy was con-
ducted, in which these suicide-bereaved families donated images
and objects related to the deceased to the project, creating an
interdisciplinary research platform between Art and Science,
which speaks a universal language (Malone et al., 2017). The
resulting artworks and mediated exhibition (Lived Lives), with
artist, scientist and the Lived Lives families, co-curated by
communities has manifested publicly both nationally and
internationally over the past decade, engaging bereaved
at-risk communities, the public (including some “at-risk” publics
– eg Travellers) and suicide prevention policy-makers, and has
facilitated dialogue, response and public action around stigma
reduction and health promotion (Malone et al., 2017). Counselling support is provided as per the original ethics
protocol. Lived Lives is embedded in the Donegal Suicide
Prevention implementation plan as part of Connecting for
Life – Ireland’s National Suicide Prevention Plan (2015–2020). Independent
evaluations
have
indicated
that
the
project
contains components consistent with a community intervention. Stigma has been succinctly defined as “a mark or sign of disgrace,
usually eliciting negative attitudes to its bearer” (Thornicroft
et al., 2007). It is a pervasive feature of mental illness which
can lead to negative discrimination. The three stigma-fuelling
elements relate to knowledge / (ignorance), attitudes / (prejudice)
and behaviour / (discrimination) (Thornicroft et al., 2007). There
are many shades of stigma including labelling and stereotyp-
ing (Lai et al., 2001). Patients with mental illness are frequently
labelled “frightening”, “unpredictable”, “dangerous” and “strange”. (Lundberg et al., 2007). Patients with depression are often
labelled “emotionally weak”, “inefficient”, “unproductive” and
“lazy” (Lai et al., 2001). Patients who experience or express
suicidal tendencies are labelled “bad”, “unbalanced” and “selfish”
(Carpiniello & Pinna, 2017). Families of patients with mental Lived Lives Paper I. Lived Lives recently manifested in a
rural Irish community which included participants who
had previous suicidal thoughts (See Lived Lives Paper I Page 3 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Paper I for images, Cleary et al., 2021). deceased woven below the portrait. The portraits are installed in
a round room chronologically according to age (with a few inter-
ruptions – see Lived Lives Paper I for images). For SPUH Lived
Lives exhibit, the Lost Portrait tapestries were installed in the
Hospital Conference Room. (Figure 4. Participants engaging Informed Consent (See Paper 1 for Image) consists of 106
signatures from families giving signed informed consent to with the Lost Portrait Gallery installed in the Conference
Room, St. Patrick’s University Hospital).
Informed Consent (See Paper 1 for Image) consists of 106
signatures from families giving signed informed consent to
Page 5 of 23 with the Lost Portrait Gallery installed in the Conference
Room, St. Patrick’s University Hospital). with the Lost Portrait Gallery installed in the Conference
Room, St. Patrick’s University Hospital).
Informed Consent (See Paper 1 for Image) consists of 106
signatures from families giving signed informed consent to Introduction and context
Depression and suicide in Ireland participate in the Lived Lives Project. (This was not installed
in SPUH due to shortage of space in the venue – see Paper I,
(Cleary et al., 2021). participate in the Lived Lives Project. (This was not installed
in SPUH due to shortage of space in the venue – see Paper I,
(Cleary et al., 2021). was to evaluate the potential of the project in an institution
providing clinical mental health care and the role this may have
in breaking the stigma around suicide and suicidal feelings in
populations engaging which such care. Following the creation of these artworks, and following the
Lived Lives families private viewing of the works and subsequent
collective approval to “go public”, a series of site-specific
experiential installations of these works and public conversations
mediated by the artist and scientist were co-created and pre-
sented with the participating families’ consent in which the
documented engagement of Lived Lives families with the
artworks became part of the artworks and with bereavement
support present. This “Lived Lives” public engagement model has
been utilized in both urban and rural community manifestations
over the past decade. (see McGuinness & Malone, EPUB,
2019). Introduction and context
Depression and suicide in Ireland 21 g consists of in excess of 92
sculpted fragments of cloth (white shirt collars) installed in
an anonymous grouping at differing heights, suspended from
invisible threads, one for each young male death that year, and
each one weighing exactly 21 g, the mythical weight of the
human soul. It is a social probe which asks questions
about our knowledge (or the lack of it) about suicide. (See The Lost Portrait Gallery consists of 39 jacquard (tapes-
try) portraits of young suicide deceased from the Lived Lives
families. These jacquards are woven, worked from dona-
tions of snapshots and memorial cards given by the families
and friends to the Lived Lives Archive. They are a photographic
representation in cloth of the deceased. Each jacquard meas-
ures 36cm × 22cm and is installed at exactly the height of
the deceased individual, with the first name and age of the Figure 1. 21g installed in Group Therapy Room, St. Patrick’s University Hospital. Figure 1. 21g installed in Group Therapy Room, St. Patrick’s University Hospital. Figure 1. 21g installed in Group Therapy Room, St. Patrick’s University Hospital. Page 4 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Figure 2. “21g -Making Stigma Visible” installed in Group Therapy Room, St. Patrick’s University: a) Lived Lives participant marking a shirt
fragment upon 21g; b) 21g shirt fragments marked with ink by Lived Lives participants, as metaphor for stigma. Figure 2. “21g -Making Stigma Visible” installed in Group Therapy Room, St. Patrick’s University: a) Lived Lives participant marking a shirt
fragment upon 21g; b) 21g shirt fragments marked with ink by Lived Lives participants, as metaphor for stigma. Figure 3. Archive Rooms, installed in the Medical Library, St. Patrick’s University Hospital. Figure 3. Archive Rooms, installed in the Medical Library, St. Patrick’s University Hospital. with the Lost Portrait Gallery installed in the Conference
Room, St. Patrick’s University Hospital). Page 5 of 23 Page 5 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Figure 4. Participants engaging with The Lost Portraits Gallery installed in the Conference Room, St. Patrick’s University
Hospital. Figure 4. Participants engaging with The Lost Portraits Gallery installed in the Conference Room, St. Patrick’s University
Hospital. ngaging with The Lost Portraits Gallery installed in the Conference Room, St. Patrick’s Universit Figure 4. Participants engaging with The Lost Portraits Gallery installed in the Conference Roo
Hospital. Methods and context This round table Confer-
ence Room conversation is transcribed and de-identified in
line with ethics protocols described above, and is reported in
Results section (see Underlying data (Lived Lives, 2021)). Consent was obtained verbally to protect anonymity and reading
assistance was offered to participants if requested. Participants
who chose to complete the feedback forms were invited to
place their completed questionnaires in the locked and secure
Lived Lives “ballot box” to further protect anonymity. Partici-
pants were not videoed during the completion of the DEQ and
Project Feedback forms (as per ethics protocol), and members
of the research team were on hand to answer any questions
relating to the feedback process. Participants were given at least
15 minute’s rest following the guided exhibition before they
completed the Feedback Form and the DEQ. These com-
l
d f
h
f
d f
h
“b ll
b
” in allied “non-clinical” but adjacent sites, where it would be
tended such that patients would not be able to “wander in” to the
exhibition (as per community installation protocol set down by
Lived Lives families). The Board was afforded an oppor-
tunity to be heard, on their own terms and expressing their
own fears of the Project (the subject matter). The Lived Lives
model has always included counselling support as part of the
project protocol. At least one bereavement counsellor was in
attendance during exhibition opening hours, as per the original
ethics approval, and per our own protocol that no one would
leave any worse than when they entered. The presence of this
counselling individual was brought to the attention of
participants at the outset of each mediated tour, indicat-
ing that support was available. This type of “safety net” has
provided reassurance to hosting institutions, and has freed up
the research team to focus exclusively on the presentation as it
is being delivered (presence), rather than having to double as
counsellors also. The Lived Lives project team travelled to the hospital for
all follow-up planning meetings. It was agreed that there
would be a “show-back” of the project engagement for hospi-
tal personnel prior to public dissemination, defined as a chance
for participants to see and comment on the data collected and
recorded (including an opportunity to withdraw consent to be
included in dissemination materials). Methods and context Building relationships and trust (Affective labour)
The methodology for working with a major psychiatric
institution in this study (Bringing Lived Lives to Swift’s
Asylum: A Psychiatric Hospital Perspective) drew on the design
from the public installation of the Lived Lives artworks in
Letterkenny and in Dunree (Lived Lives, Paper I Cleary et al., 2021)
and with The Travellers in Pavee Point (Lived Lives: A Pavee
Perspective Malone et al., 2017). It was the destination of a
Wellcome Trust-funded project titled “Lived Lives: from Tory
Island to Swift’s Asylum”, in which Lived Lives manifested in a
number of communities between Tory Island and St Patrick’s
University Hospital (McGuinness & Malone, 2019 EPUB). Collaborative pre-planning with Senior SPUH administrators
and psychiatrists began in Spring, 2017. A forum was estab-
lished, and representatives from hospital leadership invited the
Lived Lives Team into the hospital. The Lived Lives model was
presented and discussed in detail and the rationale and path-
way towards materializing Lived Lives with the psychiatric
institution Board was outlined and agreed, with its attendant
focus on establishing trust, and addressing stigma reduction. As the institution was a busy acute clinical / working site, it was
agreed from the outset that the elements of the exhibition would
not be located in any of the in-patient units, but could be situated Lived Lives Paper II. Here, in Lived Lives Paper II, we
describe bringing Lived Lives to St. Patrick’s University
Hospital (SPUH), which is Ireland’s largest and oldest psychi-
atric hospital. It was founded in 1746 through a donation from
Jonathan Swift, Dean of St Patrick’s Cathedral and author of
Gulliver’s’ Travels. In its early years it was known as “Swift’s
Asylum”. It is now a modern 302-bedded mental healthcare
facility in South inner city Dublin with national and interna-
tional expertize in the multi-disciplinary treatment of moderate
to severe mental illness and with a range of in-patient and
out-patient programmes (See St Patrick’s University Hospital
website). The objective of bringing Lived Lives to this context Page 6 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 and that no one should leave the guided exhibition “any worse
than when they entered”. Methods and context Participants were invited to ask any
questions they had in relation to the exhibition, and they were
informed that there was no onus on them to proceed to participa-
tion, or any onus to disclose whether or not they were patients at
SPUH. Hard copies of “Suicide in Ireland 2003–2008” (Malone,
2013) (publication) were available for participants to view as
an example of output(s) from the project. Participants were also
informed that the event would be documented for the purpose
of education, training, research and new knowledge and under-
standing around suicide. Participants were given the opportunity
to advise a member of the research team if they did not wish
to be documented (audio / visual), and bereavement counsel-
lors in attendance were also introduced. Participants were
also informed that the final part of the Lived Lives exhibition
would involve a reflective conversation among the artworks
(in the Hospital Conference Room) which would also be
documented (audio / visual) and participants could choose
whether or not they wished to participate in this conversation. As with previous public engagements over the past decade,
participants were informed that any recording and transcript
would be securely stored on encrypted, password-protected
hard-drives on the hospital server, backed up daily, and would
only be used for educational, training and research purposes. They were also informed these would never be used for com-
mercial purposes. Following the event, a de-identified transcript
of the Conference Room conversation was generated for the
purpose of this paper, in which any personal identifiers (such
as name), or quasi-identifiers that indirectly pointed to the
individual’s identity were deleted. Following this verbal / oral
consent procedure, consented participants were led into the
Lived Lives exhibition. When the guided tour was completed
(at the Conference Room site), the audience were then invited
to take a seat around the table and to provide their insights,
feelings and reflections, and to observe whether anyone had
regrets about joining the Exhibition. As per ethics protocol,
no onus was placed on participants to join or contribute to the
conversation. Participants were informed that the documented
outputs (audio / visual and transcript) would be used for
research, education and understanding about suicide. Attention
was drawn to available counselling support. Participants were
also invited to complete the anonymous written feedback
forms as is usual with Lived Lives. Methods and context Two video screens in the room played footage of Lived
Lives families engaging with the artworks including Archive
Rooms. The audience were invited to engage physically with this
installation and took time to do so (Figure 3). 21Grams in Group Therapy Room: Making Stigma Visible; 21Grams in Group Therapy Room: Making Stigma Visible;
Lived Lives - Garden Walk
Archive Rooms in the Medical Library
Lost Portraits Gallery in the Hospital Conference Room. Lost Portrait Gallery in The Conference Room: When this
ended, the audience were moved next door to The Conference
Room where The Lost Portrait Gallery was installed around
the walls of the room (Figure 4: Participants engaging with The
Lost Portraits Gallery installed in the Conference Room,
St. Patrick’s University Hospital). A video screen played foot-
age of Lived Lives families engaging with the Lost Portrait
Gallery. It also included a powerpoint slide of written feed-
back from one of the Lived Lives mothers as follows (The most
profound part of this Visual Arts Autopsy for me was when
I went into the circular white room…..I stood in front of her
(Fiona’s portrait) and I put my hands either side of her face. There was nobody in the world but us… … I kissed her and
walked out of the circular room and stood outside looking in at
her for what seemed like an age… …Three days later, I still feel
healing and warmth. I felt the best I have felt in 5 years since
my beloved 1st born child Fiona (aged 20) handed her life back
to God. My block of ice in my chest is thawing” (Fiona’s mother,
Galway, 2009 EPUB Page 2–4,). The Conference Room engage-
ment was the final step on the mediated tour, and participants
were invited to consent to join the team at the Conference
Room table for conversation and feedback as outlined above. Archive Rooms in the Medical Library Archive Rooms in the Medical Library Archive Rooms in the Medical Library Lost Portraits Gallery in the Hospital Conference Room. Lost Portraits Gallery in the Hospital Conference Room. Methods and context The artist invited the audience to make a fingerprint mark on
any of the shirts by dipping their finger in ink and then onto the
shirt fabric as a metaphoric means of “making stigma visible”
(Figure 2 a & b). suicide deaths annually with depression (“half the story” – the
estimate is more like at least 16 per year, with an SMR of 6.6). (All Ireland Traveller Health Study Research Team, 2012). The artist invited the audience to make a fingerprint mark on
any of the shirts by dipping their finger in ink and then onto the
shirt fabric as a metaphoric means of “making stigma visible”
(Figure 2 a & b). Attendees, participants, delegates and installations
Participants at Lived Lives SPUH was open to the public with
service users, staff and the public welcome to attend, with 86
in total engaging. The final day of the exhibition formed part
of an Annual SPUH Founders Day Conference (November
2017), which included mental health professionals, service users,
policy makers and senior administrators, and was advertised
on the hospital website and was open to the public. Fifty-eight
participants attended via the SPUH Founder Day Conference and
experienced the mediated LL Exhibition which is summarized
as follows to illustrate the process: Garden Walk: Following this engagement, the audience were
then led through the hospital garden to avoid any in-patient
exposure, and towards the very old part of the hospital build-
ing. Unintentionally, the 10 minute walk took on an element
of reverence, with a funereal likeness in the failing light, with
in-audible quiet conversations emerging from the group in
relation to the event along the way. The Clinical Director at SPUH (JVL) welcomed and introduced
the audience to Lived Lives team in SPUH Lecture Theatre;
Lived Lives team then introduced the project, and invite potential
participants to move to a room adjacent to the commencement
of the guided exhibition, where all elements of participation
are outlined to the group to obtain informed consent (oral) as
outlined above. Archive Rooms in Medical Library: Once the old part of the
hospital building was reached, the audience were moved into
the Medical Library, where Archive Rooms was installed
(Figure 4). Belongings of the deceased were carefully placed by
the artist amongst medical and psychiatric textbooks and Jour-
nals. Methods and context The process of the mediated journey of Lived Lives instal-
lations at SPUH (see SPUH mediated tour, McGuinness &
Malone, 2019 EPUB, page 64) Following the introduction to the Lived Lives team at the
Founders Day Conference, SMcG and KMM proposed to the
audience that rather than listen to another lecture, delegates
were invited to rise from their seats and move through the
hospital towards the room adjacent to the Lived Lives Exhi-
bition where informed oral / verbal consent was obtained (as
described above). Following this procedure, participants were
invited to move into Lived Lives as contributors rather than
consumers. This audience then moved with the team towards
the 1st art installation – namely 21 g at SPUH – Making
Stigma Visible. 21 g at SPUH – Making Stigma Visible: The artwork had
been installed in the Group Therapy Room (Figure 1). In this
iteration, 21 g consisted of 151 shirt fragments, which was
chosen to impart a special mental health-related significance to
depression and its treatment. As mentioned above, there are an
estimated 302 suicide deaths in Ireland annually that had
suffered from depression. This number was halved by the
artist to 151, and intended to depict “half the story”. This turn
of phrase is often used satirically when a fact / statistic is being
under-estimated. Eight shirt fragments also carried the wagon-
wheel cultural logo for Travellers on the collar to reflect traveller 21 g at SPUH – Making Stigma Visible: The artwork had
been installed in the Group Therapy Room (Figure 1). In this
iteration, 21 g consisted of 151 shirt fragments, which was
chosen to impart a special mental health-related significance to
depression and its treatment. As mentioned above, there are an
estimated 302 suicide deaths in Ireland annually that had
suffered from depression. This number was halved by the
artist to 151, and intended to depict “half the story”. This turn
of phrase is often used satirically when a fact / statistic is being
under-estimated. Eight shirt fragments also carried the wagon-
wheel cultural logo for Travellers on the collar to reflect traveller Methods and context Written approval was
obtained from St Patrick’s University Hospital Ethics Committee
for this study and as per protocol, informed verbal / oral
consent (to preserve anonymity) was obtained from all
participants prior to taking part (SPUH Protocol 22/17). [“Consent will be obtained verbally by the researcher, which
will take place after each Lived Lives tour... Questionnaires will
be left to one side for participants to take if they wish to take
part (rather than handing them out as this may lead to undue
pressure on individuals to take part”)] As per this ethically
approved protocol, participants were not videoed during the
completion of the DEQ Questionnaire or Project Feedback
Form, to preserve anonymity. Written ethics approval was also
provided by the SVUH Ethics and Medical Research Com-
mittee for an amendment to the original Lived Lives protocol
(SVUH 20/02/2006) to include Lived Lives manifesting and
being documented in SPUH, and for the completion of the
anonymous exhibition feedback form and anonymous Discrete
Emotions Questionnaire (SVUH Protocol ID 07/07/2017). As
limited literacy has been a feature of some of Lived Lives
previous iterations, it was advised and considered that verbal /
oral informed consent could address informed consent with sen-
sitivity, so as not to create any embarrassment or upset. Before
entering the guided exhibition, participants were gathered
together in a room adjacent to the exhibition, where story boards
were erected describing the history of the project. In this room,
the artist and scientist told / informed all of the prospective
participants about the exhibition, and its journey and com-
ing together of art and science for a greater understanding and
knowledge related to suicide. They were also told “what to
expect”, in-that suicide is a sensitive and sad subject, and that
the objective of the exhibition was to learn and understand
more about suicide through the guided exhibition and the
voluntary participation and voluntary feedback of participants, Page 7 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 quantitative and qualitative data were up-loaded into an
encrypted password-protected drive on the SVUH server for
analyses, which was fire-wall protected and backed up daily. suicide deaths annually with depression (“half the story” – the
estimate is more like at least 16 per year, with an SMR of 6.6). (All Ireland Traveller Health Study Research Team, 2012). Q7. The Discrete Emotions Questionnaire In total, 60 participants completed the DEQ in full (Harmon-
Jones et al., 2016). No significant effect on participants’
positive emotions was found after viewing Lived Lives, Wilks
Lambda =0.98, F (2, 58) = 0.64, p = >0.05. There was however
a significant effect on negative emotions, Wilks Lambda =
0.16, F (4, 56) = 70.25, p = < 0.05, multivariate partial eta
squared = 0.834. The means (average) and standard deviations
(how dispersed the data was from the mean) of these emotions are
presented in Table 2 below. In total, 60 participants completed the DEQ in full (Harmon-
Jones et al., 2016). No significant effect on participants’
positive emotions was found after viewing Lived Lives, Wilks
Lambda =0.98, F (2, 58) = 0.64, p = >0.05. There was however
a significant effect on negative emotions, Wilks Lambda =
0.16, F (4, 56) = 70.25, p = < 0.05, multivariate partial eta
squared = 0.834. The means (average) and standard deviations
(how dispersed the data was from the mean) of these emotions are
presented in Table 2 below. Question 7 was an adapted version of The Discrete Emotions
Questionnaire (Harmon-Jones et al., 2016) which intends
to measure self-reported discrete emotions associated with
emotional states. The scale measures 32 discrete emotions
categorised into 8 broader sub-scales (4 negative and 4 positive):
anger, fear, sadness, disgust, anxiety, desire, relaxation and
happiness. Individuals were asked “while viewing Lived Lives,
to what extent did you experience these emotions?” and then
instructed to rate each emotion from 1 (not at all) to 7 (an extreme
amount). Finally, question 8 was a qualitative question asking
participants about their overall response to the Lived Lives
works and the option to add any “further comments” relating
to the previous questions if they so wished. As has always been
the case, the Lived Lives Feedback Form was presented on pink
paper, as a satirical nod to the paperwork for involuntary
detention in a psychiatric hospital in days past, which was
known as “The Pink Form”. The questionnaire also gave participants the opportunity to
provide qualitative feedback as part of Q.8 which read: “Having viewed and experienced Lived Lives what is your
response (your immediate thoughts and feelings etc.)? Do you
have any further comments on any of the previous questions?”. Questionnaire data analysis Descriptive statistics were conducted using SPSS Version 25 to
assess demographic information about participants. Frequency
analyses were then conducted to assess the frequency
of the answers “yes”, “no” and “I don’t know” in response
to the quantitative questions (questions 1–6) across the
sample. One-way
repeated
measures
ANOVAs
were
conducted using SPSS to assess which emotions on the Discrete
Emotions Questionnaire these individuals felt most strongly
after viewing Lived Lives. ANOVAS were conducted with
the Lived Lives materials as the independent variables and
the average ratings on each emotion subscale as the depend-
ent measures. Separate ANOVAS were conducted for the posi-
tive subscales (desire, relaxation, happiness) and negative
subscales (fear, anger, sadness, disgust, anxiety), as recom-
mended by the authors of the scale (Harmon-Jones et al., 2016). A thematic analysis was conducted on the written feedback to
the open-ended question asking for further comments, based
on methods described by Braun & Clark (2006). P17 (Female, 25): “Very powerful exhibition, emotional
experience. Appreciated all the details that went into this and
the thought for the families felt strongly connected with this
subject more than ever before”. P51 (Female, 64): “Very moving and emotional. It brought the
people who died by suicide to a living state and their families
feelings, so real. Thanks for this experience.” P51 (Female, 64): “Very moving and emotional. It brought the
people who died by suicide to a living state and their families
feelings, so real. Thanks for this experience.” Beneficial: Mental health professionals who engaged with
the Lived Lives project also reported finding the experience
beneficial e.g.: “Having viewed and experienced Lived Lives what is your
response (your immediate thoughts and feelings etc.)? Do you
have any further comments on any of the previous questions?”. Sad and Glad: Many participants expressed that although
they found the exhibition sad and emotional, they were glad
they had attended as it had put the issue of suicide in perspec-
tive for them and deepened their understanding of the topic
e.g.: Evaluation In addition to written participant feedback, two independent
evaluators (Dr. Consilia Walsh - psychiatric clinician and
Dr. Aine Phillips - artist) evaluated the project by sitting in and
observing Lived Lives “in action” at SPUH before providing
written feedback, reflections and assessment. This method of
evaluation was informed by an ethnographic approach similar
to that used in previous iterations of Lived Lives (see Malone
et al., 2017) and was chosen due to the immersive and participatory Page 8 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Lives Conversation in the Conference Room; Results III reports
on the external evaluation of Lived Lives in SPUH. nature of the installation. Both evaluators (Dr. Consilia Walsh
and Dr. Aine Phillips) provided written informed consent to
publishing these evaluations. Results I: Participant questionnaire feedback Results I: Participant questionnaire feedback p
q
A total of 68 people completed the questionnaire, ranging in
age from 20–73, with a mean age of 42. The sample was 71%
female (n= 48). Four participants identified as patients, 53 iden-
tified as “not a patient” and 11 participants did not answer this
question. 17 of the participants took part in the final day of
the Lived Lives mediated exhibition as part of the Founders
Day Mental Health Conference hosted by the hospital. Participant feedback
Questionnaire data from Lived Lives at St. Patrick’s
Mental Health Services, Nov 2017. An ethically approved
anonymous semi-structure questionnaire was based on feed-
back forms used at previous Lived Lives installations, and
consisted of eight questions (see Extended data (Lived Lives,
2021)). There were also spaces provided to record demographic
details, patient status and the completion date. The first three
questions were directly related to the participants’ thoughts on
Lived Lives and the next three questions were related to their
own personal experience of suicide bereavement and mental
health history (including suicidal feelings). Participants’ responses to questions 1–6 about their engagement
with Lived Lives are presented in Table 1. Results Results I reports on the written feedback (quantitative and
qualitative) provided by 68 attendees over the 3 day course
of the Lived Lives event; Results II reports on the taped Lived P4 (Female 25): “Excellent exhibition. A very emotional
experience, beneficial for everybody to see. Particularly relevant Page 9 of 23 Page 9 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Table 1. Lived Lives at SPUH - feedback form
responses. Q1. Do you think Lived Lives could benefit people
following a suicide death? Answer
Frequency (n)
%
Yes
62
91
No
-
-
Don’t know
5
7.5
Missing
1
1.5
Q2. Do you think Lived Lives could somehow
benefit people with mental health difficulties? Answer
Frequency (n)
%
Yes
60
88
No
1
1.5
Don’t know
6
9
Missing
1
1.5
Q3. Do you think Lived Lives “glamorizes” and/or
“romanticizes” suicide? Answer
Frequency (n)
%
Yes
-
-
No
68
100
Don’t know
-
-
Missing
-
-
Q4. Have you lost anyone to suicide? Answer
Frequency (n)
%
Yes
42
62
No
25
37
Missing
1
1
Q5. Have you personally experienced any mental
health difficulties? Answer
Frequency (n)
%
Yes
31
46
No
36
53
Missing
1
1
Q6. Have you ever experienced suicidal feelings? Answer
Frequency (n)
%
Yes
24
35
No
43
63
Mi
i
1
2 If yes, do you think the Lived Lives project/
exhibition could reduce suicidal feelings? Answer
Frequency (n)
%
Yes
31
46
No
5
7
Don’t know
1
1 Q1. Do you think Lived Lives could benefit people
following a suicide death? Answer
Frequency (n)
%
Yes
62
91
No
-
-
Don’t know
5
7.5
Missing
1
1.5 Table 2. Discrete emotions questionnaire responses. Table 2. Discrete emotions questionnaire responses. Negative Subscales (items)
Mean
SD
N
Anger
(anger, rage, mad, pissed off)
1.77
1.14
60
Disgust
(grossed out, nausea, sickened, revulsion)
1.56
0.87
60
Fear
(terror, scared, panic, fear)
1.96
1.25
60
Anxiety
(dread, anxiety, nervous, worry)
2.64
1.20
60
Sadness
(sad, lonely, grief, empty)
4.12
0.94
60 Q2. Do you think Lived Lives could somehow
benefit people with mental health difficulties? for healthcare professionals in psychiatry as promotes greater
understanding of the effects of suicide on victims and
their families. Results Feel I have greater empathy for victims and
families” for healthcare professionals in psychiatry as promotes greater
understanding of the effects of suicide on victims and
their families. Feel I have greater empathy for victims and
families” P53 (Female, 53): “Professionally I feel reenergised and
refocused on the work needed to do to prevent suicide and
reduce the stigma that families and loved ones experience. It
is left me with a heightened of responsibility. Finally as an
individual it has reinforced the need for compassion for those
who struggle within my personal social network and beyond. Thank you so much for your gift of this experience “Lived
Lives”. Perhaps it will come back to for another time “enriched
post New York” and post APA!” Q5. Have you personally experienced any mental
health difficulties? Also, because I’ve been there myself but so
gracious to be still here” For M2, a service user, the Lost Portrait Gallery was pow-
erful in a different way: “there is something quite powerful
about the fact that we are here – in this room, and statisti-
cally I could be on the wall and I’m not, and I’m very grate-
ful I’m not and that gives me encouragement and hope”. F3,
another service user, echoed this sense of gratitude saying, “in
my own past I have attempted suicide, and I feel a profound
sense of gratitude actually at being here that I’m not up on the
wall”. P63 (Female, 44): “Just a profound sense of humanity you have
brought to the lives represented in the exhibition – and to the
lives of people who commit suicide and their families. And
gratitude. Overwhelming gratitude at my life and loves.” P35 (Female, 32): “I am grateful for the organisers, feel this
exhibition needs to be viewed by a lot more people, it is so
interesting maybe some answers beyond this project and feel a
lot is to be said about the lives behind suicide #Mindblowing
Thank you” P35 (Female, 32): “I am grateful for the organisers, feel this
exhibition needs to be viewed by a lot more people, it is so
interesting maybe some answers beyond this project and feel a
lot is to be said about the lives behind suicide #Mindblowing
Thank you” Any regrets about participating?: When asked if they had
any regrets about attending the event, none were expressed by
the participants. F1, psychiatric trainee, expressed no regrets and
said “I think I have to say I am even struck by how brave people
are about sharing their own personal experiences even in
this room, and I think it’s kind of amazing in terms of the power
of this exhibition that people feel – open enough and they
feel there is a safe enough space to talk about their own experi-
ences. It reminds me of why I became a doctor”. F4, a senior
employee in the hospital, said that despite reservations about
“crying in front of everyone” before she came to the exhibi-
tion, she had no regrets after participating “with these young
people in front of you (tapestries) who lost their lives in this
way, you wouldn’t be human if you didn’t cry. Q5. Have you personally experienced any mental
health difficulties? I don’t mind
who sees me cry”. F5, another senior employee at the hospital
echoed that she had no regrets in participating and she discussed
the considerations the organisation had to make in advance
of collaborating with the Lived Lives project: “We actually
had to give it considerable consideration, because obviously
you can see here today, the vulnerability, the humanity
that we’ve all experienced, professional or untrained you know,
its opened up something and we had to take that very seri-
ously, but I am so so glad we did”. One participant (P26),
whilst calling the project “incredible research”, raised an
objection to advance notice not being provided regarding
video-documenting, and to the nature of how consent to
participate was obtained. Her view was acknowledged by
the team who reiterated the consent procedure and the verbal
notice on documenting the event before the commencement of
the mediated tour (and the participant subsequently remained to
the end of the mediated tour). Referring to the invitation as part Bereavement support is always provided, and is on-hand
throughout the event. The research team offered company to
those who seemed to be visiting alone. As it happened, the
majority of participants had at least one other participant with
them. In reality, this support was more of offering companion-
ship, “a quiet word” and personal chaperoning if this offer was
taken up. A debriefing session with our Art Therapist for
younger participants seemed to be popular and effective (see
McGuinness & Malone, 2019, Closings Sessions. EPUB Page 55). Results II: Lived Lives Conference Room Conversation and
participant verbal feedback amongst Lost Portrait Gallery
(N=17) (McGuinness & Malone, 2019, SPUH Conference
Room Conversation, EPUB, page 65) (see also Underlying data
(Lived Lives, 2021)). Results II: Lived Lives Conference Room Conversation and
participant verbal feedback amongst Lost Portrait Gallery
(N=17) (McGuinness & Malone, 2019, SPUH Conference
Room Conversation, EPUB, page 65) (see also Underlying data
(Lived Lives, 2021)). 17 attendees (12 females, 5 males) from the Mental Health
Conference participated in the Lived Lives Conference Room
Conversation. They included service users, mental health
professionals and senior hospital administrators. 17 attendees (12 females, 5 males) from the Mental Health
Conference participated in the Lived Lives Conference Room
Conversation. They included service users, mental health
professionals and senior hospital administrators. Q5. Have you personally experienced any mental
health difficulties? P61 (Female, 41): “A very valuable piece of work for staff work-
ing in mental health, for patients (+ ex-patients), for family
coping with grief.” Young people should see: Many participants felt that they
exhibition should be seen by many more people, in particular
young people e.g.: P13 (Male, 26): “It’s an exhibition all young students should
be opened to in secondary schools”. Page 10 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 P21: (Female, 40): “I think everyone should see this. I wish for
all teenagers to see this + talk about suicide + what it means. I think it is really valuable work + I’m glad to have been a part
of that, even though it was painful at times + difficult” described the exhibition as “profoundly human” and described
it as “really powerful to be allowed feel”. F1 also highlighted
the significance of the Lost Portrait Gallery being installed in
the Hospital Conference Room “where lots of important deci-
sions are made”, saying “I imagine it’d have been quite stark for
the people who had meetings in here this week.” F4 and F5,
another two employees at the hospital, also remarked on the
“contrast” of the project being in a psychiatric hospital. F4
described the experience as leading her to reflect on her work
and she described how speaking about the issue as part of the
exhibition had made her “a bit more hopeful”. Similarly, F5, An
Administration Director at the hospital, reflected on an earlier
conversation about professional failure in terms of suicide deaths
saying “out of a sense of failure there is a power in the vulner-
ability. It can be very empowering to experience something
like this and re-invigorate our passion to make a difference”. Thanks to the Lived Lives research families: The qualitative
space was also used by many participants to express their
gratitude – for their own lives and to the Lived Lives families
for sharing their stories and to the organisers of the exhibition
for providing the opportunity for them to engage with
the Lived Lives project e.g.: P54 (Female, 51): “Gratitude to the artist, & the families. They have been so brave and courageous. Sadness for all the
lives lost and the loved ones left. Pain for the friends lost and
their families. Siting of Lived Lives artworks in and within the
psychiatric hospital St Patricks is recognized as a leading and respected
advocate for improving awareness of mental health issues in
Ireland and Internationally. Since its founding, the organisation
has been at the forefront of seeking to reduce mental health
stigma and misunderstanding of mental health difficulties in
society. The collaborative proposal brought Lived Lives into St. Patrick’s
University Hospital. Together with backing from the Wellcome
Trust, and consistent with their reputation as advanced lead-
ers in mental healthcare, (and respectful of the core values of
Swift’s vision), and following careful consideration, the Hos-
pital Board elected to take on the project. There was enormous
symbolism, meaning and irony in the placement of the artworks
within the psychiatric hospital, which was thought-provoking
for the participants, and heightened the impact of the Lived
Lives message of confronting stigma. Some participants chose to
reflect on the Archive Room being placed within the Medical
Library as a way to raise the question about suicide and depres-
sion “What do we really know”, and how the Archive Room
contents presented a deep and different knowledge. One par-
ticipant was prompted to speak of the challenges associated
with divestment in the aftermath of a suicide loss of a loved one,
and what one does with their belongings as they are so emo-
tionally charged, observing that the Lived Lives experience
and especially Archive Rooms, helped her to “overcome this”,
and see things in a different way. Others found the placement of the Lost Portrait Gallery
surrounding the Conference Room walls as particularly poign-
ant, gently but persuasively questioning the authorities who
might have attended the Conference Room on hospital busi-
ness during the week of Lived Lives about the problem of
suicide deaths. Others found the installation of 21g in the
Group Therapy Room as particularly haunting “If I was to
come to this exhibition again, I’d go back to the shirts every
time”. Many but not all welcomed the invitation to place a mark
“stain” on the shirt fragments as a metaphor for making
stigma “visible”. Indeed, one participant shared that he wasn’t
sure if he wished to wipe the ink from his thumb, it being a
reminder of the profound feeling he had experienced by mak-
ing a mark. Q5. Have you personally experienced any mental
health difficulties? Immediate feelings: While discussing their immediate feelings
following engagement with Lived Lives, many participants
agreed they would describe it as “powerful”. M1, a member
of the general public, said “its powerful, that’s all I can say. It
makes you want to make a difference when you leave this
room”. Another participant, a psychiatric trainee in the hospital
(F1), described the conflict she has experienced between
grieving for those lost by suicide and remaining professional. She Page 11 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 and reflecting through writing. To this end, both drew on their
wealth of professional experience in their respected fields of
clinical science and art. The brief summary of their evaluations
provided here foregrounds their most salient points (see full
texts in Underlying data (Lived Lives, 2021)). Objective feed-
back from the senior clinician psychiatrist (CW) drew attention to
“the journey” aspect of the event, beginning with the anonymous
shirt collars of 21g, moving on into Archive Room, where
you get to know the deceased a little better, and then into The
Lost Portrait Gallery, where “identity” is revealed. CW also
articulated how Lived Lives communicates the sensitivity around
suicide and suicide bereavement in a very different and tangi-
ble way. As a clinician, she has been dealing with suicide and
attempted suicide all her clinical life, and found the Lived
Lives project touched and moved her in a new and compelling
way. She also reflected on how this form of communication
could resonate so well with young people, encouraging them to
speak out as they did in the videos. Artist AP chose to reflect on
aesthetics and meaning. For example, she sees 21g as a “throng of
anthropomorphic collars which gives comfort in its evocation
of unity”, suggesting that the collars are “spectral traces of
the lost individual lives”, representing succour or solace in the
subtle beauty and tactile substance of the artwork”. “Viewers”,
she says, are encouraged to touch the collars, “with ink stained
thumbs to leave their own printed trace upon the installation”. This act of touching she observes denotes “physical contact
which in turn enables a psychological touching to simultaneously
happen, allowing the participant to intellectually and emotion-
ally grasp this complex subject”. Q5. Have you personally experienced any mental
health difficulties? AP suggests that Lived Lives
“gives audiences access to shared authorship in the work and
the means to make a difference in the way the subject of suicide
is mediated and disseminated. McGuinness and Malone have
enabled the participating families to define and direct how
their loved one’s stories are depicted in the project so this
relational approach was maintained throughout”. AP observes that
the project explores and reveals layers of invisible knowledge -
private thoughts and feelings of suicide-bereaved individuals are
given public disclosure within a supported, safe and imaginative
context. This she says allows for the dissolving of the private
and public in therapeutic ways to help dismantle stigma. She
says that Lived Lives is a collective experience, often in silence
and contemplation alongside dialogue and interaction and
this provides for a necessary and effective processing of the
subject to take place. AP concludes that the Lived Lives authors
invite us to align our deeper selves with the artworks and, via
the art, we are helped to address suicide as a meaningful life
experience. She says that the work does not “deny our tragedies”,
but becomes “a legitimate place for solemn emotions and creates
a dignified home for both sorrow and hope” (see Underlying
data for complete CW and AP evaluations (Lived Lives, 2021);
for other evaluations to date see McGuinness & Malone, 2019,
EPUB Page 68–96). A show-back of “Lived Lives
A psychiatric hospital per- of Making Stigma Visible to mark his thumb in ink and leave a
mark on one of the 21 grams white shirts and “make stigma
visible”, M3, an addictions counsellor, said “I feel privileged to
be part of it you know. I don’t know if I want to wash this off
or not you know. You know its’ powerful”. Siting of Lived Lives artworks in and within the
psychiatric hospital The Garden Walk also created a quiet space in
the fading light for reflecting as participants moved from 21g
Making Stigma Visible to the remainder of the exhibition in
the older part of the hospital. A show-back of “Lived Lives, A psychiatric hospital per-
spective” was arranged for six weeks following the event in
SPUH, with notably favourable feedback from attendees, and
with no objections expressed towards pursuing its dissemination
for education, training and research. Results III: Clinician and artist independent evaluation
(Dr. Consilia Walsh, Dr. Aine Phillips)
A priori, both evaluators elected to assume an ethnographic
approach to the Lived Lives experience - observing, engaging Discussion Transposing a dynamic, mediated, interactive and collaborative
science-arts project about suicide into a psychiatric institu-
tion was a ground-breaking experience, which to our knowledge
has not previously been attempted. Our findings identified a
deep stigma that still surrounds mental illness and everything
associated with it. In this study and in their own words, those
who serve to support and treat all kinds of mental illness,
witness and experience stigma in their daily work. When the
stigma of suicidal thoughts and fatal deeds is added to the
stigma of mental illness and the stigma of having been in a
psychiatric hospital, it seems to create a pervasive muting
of voice and a response of recoil and intervention paralysis. Situated within the Lived Lives project lies a catalyst for
reconciliation and change, where an aperture for voice is cre-
ated through a transformative process attendant in the artworks
and facilitated through the mediated safe-space experience. Lived Lives sits comfortably in discomfort, unafraid to call
out the home-truths about stigma and its pervasive and
pernicious impact, managing to do so with deepest respect and
empowerment, and with restoring identity at its core. There are limitations with regard to aspects of the study. Advertising for such an event is challenging. The event was
posted on the hospital website where it was stated that it was
open to the public. Perhaps the fears of visiting an event
hosted in a psychiatric institution were a stigma-related deter-
rent. One way or another there are barriers to reaching the
public and attracting them to participate in such projects
around suicide and stigma (Knaak et al., 2017). Previous
iterations of Lived Lives in rural communities have also had
a mixed public response, where media interviews with scien-
tist and artist on local radio stations have proven to be effective
(McGuinness & Malone, 2019), EPUB page 99). Future research
on what factors may influence attendance and the thoughts
of those who choose not to attend such projects would be
beneficial and enlightening in this regard. There may be an
opportunity via evaluations of future iterations of Lived Lives
to evaluate the process from its inception phase which would
include liaising with the wider community to increase engage-
ment and reach to identify and potential avenues in this regard. Discussion We acknowledge that the use of the DEQ to measure partici-
pants’ emotional responses has its limitations in terms of its
cross-sectional nature and inferences in terms of any longer-term
impacts of participating in the project on emotions or indeed
behavior cannot be made. As it is a relatively new instrument,
the DEQ itself requires further research in different contexts
(Harmon-Jones et al., 2016). Our previous Lived Lives iteration in Dunree, Co. Donegal
demonstrated that Lived Lives could work well with people with
previous suicidal experiences. Cleary et al., 2021 (Lived Lives
Paper I). Many of the participants in the Lived Lives SPUH
project acknowledged their own suicidal feelings, previous
suicidal feelings and acts, and the experience of suicide
bereavements, with an affirmative and sharing disclosure to the
Conference Room conversation. No participant thought the Lived Lives experience glamor-
ized, glorified, romanticized or fantasized suicide, with many
stating the opposite, as it focusses on the harsh reality for the
deceased and the bereaved. As one of the Lived Lives fami-
lies bluntly put it, “dead is dead” (Suzanne – Galway, 2009. see
EPUB Page 2–4). In addition to the evaluations by an independent scientist
and artist in this paper, multi-source and multi-disciplinary
evaluation of the Lived Lives Project has been provided by
six further diverse sources of expertize over the past decade,
embracing science arts and humanities, bearing witness and
sharing their testament and testimony (McGuinness & Malone,
2019; EPUB, page 68–96). Their guiding and generous insights
provide a deep, grounding and enduring critique, with an inher-
ent value to the project being greater than the sum of their
parts. As such, all these evaluations need no introduction,
translation or interpretation, and speak for themselves as
they enrich the project. In this vein, both evaluators (Walsh
and Phillips) provided particular insights, reflection and
understanding (see Underlying data (Lived Lives, 2021)). The DEQ was included to assess emotional responses
following the engagement with Lived Lives. Consistent with our
expectation, the participants reported nothing positive about the
emotional response, but instead experienced significant nega-
tive emotional feelings consistent with a “visceral response”
frequently reported by participants. Many participants describe the
experience as “powerful”. Results III: Clinician and artist independent evaluation
(Dr. Consilia Walsh, Dr. Aine Phillips) A priori, both evaluators elected to assume an ethnographic
approach to the Lived Lives experience - observing, engaging Page 12 of 23 Page 12 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 the experiences of patients and their families as they relate
to stigma, as these represent the core start and end points of
any anti-stigma effort or program. Collaboration, Curiosity,
Optimism and Experimentation have kept the project fresh,
relevant, relatable and nimble, and evaluation is integral to
the process. Lived Lives has been described as a community
intervention (see Christabel Owens evaluation and Figure, in
Malone et al., 2017), and could be considered “an intervention
prototype” (Ungar et al., 2016). Extended data Irish Social Science Data Archive: Lived Lives, https://www. ucd.ie/issda/data/livedlives/ (Lived Lives, 2021). Study number
(SN): 0070-00 This project contains the following underlying data:
-
Lived Lives SPUH Blank Questionnaire (Copy of
questionnaire used at Lived Lives SPUH) Data availability
Underlying data
Irish Social Science Data Archive: Lived Lives, https://www. ucd.ie/issda/data/livedlives/ (Lived Lives, 2021). Study number
(SN): 0070-00 Stigma. Front Psychiatry. 2017; 8: 35.
PubMed Abstract | Publisher Full Text | Free Full Text
Central Statistics Office: Vital Statistics Statbank. 2019; accessed: 11/11/2019.
Cleary E, Malone KM, Corry C, et al.: Lived Lives at Fort Dunree: A rural
community perspective [version 1; peer review: 1 approved, 1 approved
with reservations]. 2021.
Publisher Full Text
Conwell Y, Duberstein PR, Cox C, et al.: Relationships of age and axis I
diagnoses in victims of completed suicide: a psychological autopsy study.
Am J Psychiatry. 1996; 153(8): 1001–1008.
PubMed Abstract | Publisher Full Text
Corrigan PW, Miller FE: Shame, blame, and contamination: A review of the
impact of mental illness stigma on family members. J Ment Health. 2004;
13(6): 537–548.
Publisher Full Text
Foster T, Gillespie K, McCelland R: Mental disorders and suicide in Northern Discussion The Lived Lives project contains elements of contemporary
anti-stigma / stigma-reduction strategies (Knaak et al., 2014;
Knaak et al., 2017) such as Ecological Validity, where our
target audiences are working partners from the outset and are
set in the richness of their natural environment. The Lived
Lives families are co-curators with Lived Experience from the
outset. Human Design contains Human Factors, Empathic
Design and Design Thinking, all of which are represented in
the conceptualization, construction, realization, manifestation
and delivery of Lived Lives. Human Centeredness and Empathy
informs the Lived Lives process and is essential for understanding The triple stigma of mental illness, suicidal experiences and
having been in a psychiatric hospital has never been quantified. It is unknown if there is an additive, cumulative or multiplier
effect of each. Clinically, these stigmas, both internal and
external. impact on self-esteem and impede recovery, which Page 13 of 23 Page 13 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 -
Lived_Lives_Paper2_SPUH_Qualitative_Feedback
(Word file containing quantitative data collected
at Lived Lives SPUH)
-
Lived_Lives_Walsh_and_Phillips_Feedback_and
Evaluation (Word document containing evaluations
of Lived Lives by Dr. Consilia Walsh and Dr Áine
Phillips) Lived_Lives_Paper2_SPUH_Qualitative_Feedback
(Word file containing quantitative data collected
at Lived Lives SPUH) warrants further research (Moses, 2014; Rüsch et al., 2005). Projects such as Lived Lives have the potential to meet stigma
head-on, and can operate within the context of suicide interven-
tion and prevention planning (as evidenced with Lived Lives
in Donegal (EPUB Pages 59–63). Lived Lives has existed in the
public domain since 2010. Over 2,000 members of the public
have engaged and interacted directly with the Project, with at
least 20% of those being young adults. Also included are
1st responders such as police and coastguard, LGBT, Irish
Travellers, Health policy makers, health providers and service
users. Many others have interacted with the project through
Lived Lives Lectures and Conversations, such as over 2,000
University College Dublin medical students – (the “doctors
of tomorrow”). No adverse events have been reported to date,
whereas an overwhelming majority of participants have shared
their positive testimony either orally, or written or both, and
many have returned for repeat engagement with the project. Lived Lives in this instance was experienced in a psychiatric
hospital as authentic at a deep level of humanity. Discussion The challenge
is to move it forward for greater exposure in communities
nationally and internationally, and further into psychoeducation
and anti-stigma whilst preserving its essence. warrants further research (Moses, 2014; Rüsch et al., 2005). Projects such as Lived Lives have the potential to meet stigma
head-on, and can operate within the context of suicide interven-
tion and prevention planning (as evidenced with Lived Lives
in Donegal (EPUB Pages 59–63). Lived Lives has existed in the
public domain since 2010. Over 2,000 members of the public
have engaged and interacted directly with the Project, with at
least 20% of those being young adults. Also included are
1st responders such as police and coastguard, LGBT, Irish
Travellers, Health policy makers, health providers and service
users. Many others have interacted with the project through
Lived Lives Lectures and Conversations, such as over 2,000
University College Dublin medical students – (the “doctors
of tomorrow”). No adverse events have been reported to date,
whereas an overwhelming majority of participants have shared
their positive testimony either orally, or written or both, and
many have returned for repeat engagement with the project. Lived Lives in this instance was experienced in a psychiatric
hospital as authentic at a deep level of humanity. The challenge
is to move it forward for greater exposure in communities
nationally and internationally, and further into psychoeducation
and anti-stigma whilst preserving its essence. These data are under restricted access due to the sensitivity
of the subject material. To access the data, please complete a
ISSDA Data Request Form for Research Purposes, sign it, and
send it to ISSDA by email (issda@ucd.ie). Researchers will be
asked to provide a description for the intended use of the data
and will be asked to agree to the terms of use, as outlined in the
request form. Data access will be granted for teaching and
research purposes under ISSDA terms and conditions. Underlying data Underlying data Irish Social Science Data Archive: Lived Lives, https://www. ucd.ie/issda/data/livedlives/ (Lived Lives, 2021). Study number
(SN): 0070-00 Data are available under the terms of the Creative Commons
Attribution 4.0 International license (CC-BY 4.0). This project contains the following underlying data: This project contains the following underlying data: -
Lived_Lives_Quantitative_Data_Paper2_SPUH
(Excel
file containing quantitative data collected at Lived
Lives SPUH) Acknowledgements
Presented in part at the 146th Annual International Scientific
Conference of the American Psychiatric Association, San
Francisco, USA. (May 2019), and at The International Academy
for Suicide Research (IASR) / American Foundation for
Suicide Prevention (AFSP) International Summit on Suicide
Research, Miami Florida, USA. (October 2019). Acknowledgements
Presented in part at the 146th Annual International Scientific
Conference of the American Psychiatric Association, San
Francisco, USA. (May 2019), and at The International Academy
for Suicide Research (IASR) / American Foundation for
Suicide Prevention (AFSP) International Summit on Suicide
Research, Miami Florida, USA. (October 2019). -
Lived_Lives_Conference_Room_Transcript_Paper2_
SPUH (Word file containing anonymised transcrip-
tion of post-exhibition interactive conversation with
the Lived Lives team) References References Stigma. Front Psychiatry. 2017; 8: 35. PubMed Abstract | Publisher Full Text | Free Full Text
Central Statistics Office: Vital Statistics Statbank. 2019; accessed: 11/11/2019. Cleary E, Malone KM, Corry C, et al.: Lived Lives at Fort Dunree: A rural
community perspective [version 1; peer review: 1 approved, 1 approved
with reservations]. 2021. Publisher Full Text
Conwell Y, Duberstein PR, Cox C, et al.: Relationships of age and axis I
diagnoses in victims of completed suicide: a psychological autopsy study. Am J Psychiatry. 1996; 153(8): 1001–1008. PubMed Abstract | Publisher Full Text
Corrigan PW, Miller FE: Shame, blame, and contamination: A review of the
impact of mental illness stigma on family members. J Ment Health. 2004;
13(6): 537–548. Publisher Full Text
Foster T, Gillespie K, McCelland R: Mental disorders and suicide in Northern All-Ireland Traveller Health Study Research Team: The health care experiences
of Travellers compared to the general population: the All-Ireland Traveller
Health Study. J Health Serv Res Policy. 2012; 17(3): 173–180. PubMed Abstract | Publisher Full Text
Bostwick JM, Pabbati C, Geske JR, et al.: Suicide Attempt as a Risk Factor for
Completed Suicide: Even More Lethal Than We Knew. Am J Psychiatry. 2016;
173(11): 1094–1100. PubMed Abstract | Publisher Full Text | Free Full Text
Braun V, Clarke V: Using thematic analysis in psychology. Qualitative Research
Psychology. 2006; 3(2): 77–101. Publisher Full Text
Brodsky BS, Spruch-Feiner A, Stanley B: The zero suicide model: Applying
evidence-based suicide prevention practices to clinical care. Front Psychiatry. 2018; 9: 33. PubMed Abstract | Publisher Full Text | Free Full Text
Carpiniello B, Pinna F: The Reciprocal Relationship between Suicidality and Stigma. Front Psychiatry. 2017; 8: 35. PubMed Abstract | Publisher Full Text | Free Full Text
Central Statistics Office: Vital Statistics Statbank. 2019; accessed: 11/11/2019. Cleary E, Malone KM, Corry C, et al.: Lived Lives at Fort Dunree: A rural
community perspective [version 1; peer review: 1 approved, 1 approved
with reservations]. 2021. Publisher Full Text
Conwell Y, Duberstein PR, Cox C, et al.: Relationships of age and axis I
diagnoses in victims of completed suicide: a psychological autopsy study. Am J Psychiatry. 1996; 153(8): 1001–1008. PubMed Abstract | Publisher Full Text
Corrigan PW, Miller FE: Shame, blame, and contamination: A review of the
impact of mental illness stigma on family members. J Ment Health. 2004;
13(6): 537–548. All-Ireland Traveller Health Study Research Team: The health care experiences
of Travellers compared to the general population: the All-Ireland Traveller
Health Study. J Health Serv Res Policy. 2012; 17(3): 173–180.
PubMed Abstract | Publisher Full Text
Bostwick JM, Pabbati C, Geske JR, et al.: Suicide Attempt as a Risk Factor for
Completed Suicide: Even More Lethal Than We Knew. Am J Psychiatry. 2016;
173(11): 1094–1100.
PubMed Abstract | Publisher Full Text | Free Full Text
Braun V, Clarke V: Using thematic analysis in psychology. Qualitative Research
Psychology. 2006; 3(2): 77–101.
Publisher Full Text
Brodsky BS, Spruch-Feiner A, Stanley B: The zero suicide model: Applying
evidence-based suicide prevention practices to clinical care. Front Psychiatry.
2018; 9: 33.
PubMed Abstract | Publisher Full Text | Free Full Text
Carpiniello B, Pinna F: The Reciprocal Relationship between Suicidality and Reference Source Möller HJ, Henkel V: What are the most effective diagnostic and therapeutic
strategies for the management of depression in specialist care? Copenhagen, WHO Regional Office for Europe (Health Evidence Network
report; 2005; accessed November 2019). PubMed Abstract | Publisher Full Text PubMed Abstract | Publisher Full Text Knaak S, Mantler E, Szeto A: Mental illness-related stigma in healthcare:
Barriers to access and care and evidence-based solutions. Health Manage
Forum. 2017; 30(2): 111–116. Knaak S, Mantler E, Szeto A: Mental illness-related stigma in healthcare:
Barriers to access and care and evidence-based solutions. Health Manage
Forum. 2017; 30(2): 111–116. Moses T: Determinants of mental illness stigma for adolescents discharged
from psychiatric hospitalization. Soc Sci Med. 2014; 109: 26–34. b
d
b
|
bli h
ll Knaak S, Modgill G, Patten SB: Key ingredients of anti-stigma programs
for health care providers: a data synthesis of evaluative studies. Can J
Psychiatry. 2014; 59(10 Suppl 1): S19–26. Phelan JC, Bromet EJ, Link BG: Psychiatric Illness and Family Stigma. Schiz
Bull. 1998; 24(1): 115–126. Lai YM, Hong CP, Chee CY: Stigma of mental illness. Singapore Med J. 2001;
42(3): 111–4. PubMed Abstract | Publisher Full Text Rüsch N, Angermeyer MC, Corrigan PW: Mental illness stigma: Concepts,
consequences, and initiatives to reduce stigma. Eur Psychiatry. 2005; 20(8):
529–539. Rüsch N, Angermeyer MC, Corrigan PW: Mental illness stigma: Concepts,
consequences, and initiatives to reduce stigma. Eur Psychiatry. 2005; 20(8):
529–539. Link BG, Cullen FT, Frank J, et al.: The social rejection of former mental
patients: Understanding why labels matter. Am J Social. 1987; 92(6):
1461–1500. Thornicroft G, Rose D, Kassam A, et al.: Stigma: ignorance, prejudice or
discrimination? Br J Psychiatry. 2007; 190: 192–193. References Publisher Full Text
Foster T, Gillespie K, McCelland R: Mental disorders and suicide in Northern
Page 14 of 23 All-Ireland Traveller Health Study Research Team: The health care experiences
of Travellers compared to the general population: the All-Ireland Traveller
Health Study. J Health Serv Res Policy. 2012; 17(3): 173–180. PubMed Abstract | Publisher Full Text
Bostwick JM, Pabbati C, Geske JR, et al.: Suicide Attempt as a Risk Factor for
Completed Suicide: Even More Lethal Than We Knew. Am J Psychiatry. 2016;
173(11): 1094–1100. PubMed Abstract | Publisher Full Text | Free Full Text
Braun V, Clarke V: Using thematic analysis in psychology. Qualitative Research
Psychology. 2006; 3(2): 77–101. Publisher Full Text
Brodsky BS, Spruch-Feiner A, Stanley B: The zero suicide model: Applying
evidence-based suicide prevention practices to clinical care. Front Psychiatry. 2018; 9: 33. PubMed Abstract | Publisher Full Text | Free Full Text
Carpiniello B, Pinna F: The Reciprocal Relationship between Suicidality and Bostwick JM, Pabbati C, Geske JR, et al.: Suicide Attempt as a Risk Factor for
Completed Suicide: Even More Lethal Than We Knew. Am J Psychiatry. 2016;
173(11): 1094–1100. Braun V, Clarke V: Using thematic analysis in psychology. Qualitative Research
Psychology. 2006; 3(2): 77–101. Brodsky BS, Spruch-Feiner A, Stanley B: The zero suicide model: Applying
evidence-based suicide prevention practices to clinical care. Front Psychiatry. 2018; 9: 33. Corrigan PW, Miller FE: Shame, blame, and contamination: A review of the
impact of mental illness stigma on family members. J Ment Health. 2004;
13(6): 537–548. Corrigan PW, Miller FE: Shame, blame, and contamination: A review of the
impact of mental illness stigma on family members. J Ment Health. 2004;
13(6): 537–548. Publisher Full Text Carpiniello B, Pinna F: The Reciprocal Relationship between Suicidality and Foster T, Gillespie K, McCelland R: Mental disorders and suicide in Northern Foster T, Gillespie K, McCelland R: Mental disorders and suicide in Northern Foster T, Gillespie K, McCelland R: Mental disorders and suicide in Northern Page 14 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 ellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Ireland. Br J Psychiatry. 1997; 170: 447–52. PubMed Abstract | Publisher Full Text
Harmon-Jones C, Bastian B, Harmon-Jones E: The Discrete Emotions
Questionnaire: A New Tool for Measuring State Self-Reported Emotions. PLoS One. 2016; 11(8): e0159915. References PubMed Abstract | Publisher Full Text | Free Full Text
Harwood D, Hawton K, Hope T, et al.: Psychiatric disorder and personality
factors associated with suicide in older people: a descriptive and case-
control study. Int J Geriatr Psychiatry. 2001; 16(2): 155–165. PubMed Abstract | Publisher Full Text
Hasin DS, Sarvet AL, Meyers JL, et al.: Epidemiology of adult DSM-5 major
depressive disorder and its specifiers in the United States. JAMA psychiatry. 2018; 75(4): 336–346. PubMed Abstract | Publisher Full Text | Free Full Text
Henderson C, Noblett J, Parke H, et al.: Mental health-related stigma in
health care and mental health-care settings. Lancet Psychiatry. 2014; 1(6):
467–82. PubMed Abstract | Publisher Full Text
Henriksson MM, Aro HM, Marttunen MJ, et al.: Mental disorders and
comorbidity in suicide. Am J Psychiatry. 1993; 150(6): 935–940. PubMed Abstract | Publisher Full Text
Knaak S, Mantler E, Szeto A: Mental illness-related stigma in healthcare:
Barriers to access and care and evidence-based solutions. Health Manage
Forum. 2017; 30(2): 111–116. PubMed Abstract | Publisher Full Text | Free Full Text
Knaak S, Modgill G, Patten SB: Key ingredients of anti-stigma programs
for health care providers: a data synthesis of evaluative studies. Can J
Psychiatry. 2014; 59(10 Suppl 1): S19–26. PubMed Abstract | Publisher Full Text | Free Full Text
Lai YM, Hong CP, Chee CY: Stigma of mental illness. Singapore Med J. 2001;
42(3): 111–4. PubMed Abstract
Link BG, Cullen FT, Frank J, et al.: The social rejection of former mental
patients: Understanding why labels matter. Am J Social. 1987; 92(6):
1461–1500. Publisher Full Text
Lived Lives: Lived Lives I : A Rural Perspective; Lived Lives II: A Psychiatric
Hospital Perspective, 2020. [dataset]. Version 1. Irish Social Science Data
Archive. 2021. Lundberg B, Hansson L, Wentz E, et al.: Sociodemographic and clinical factors
related to devaluation/discrimination and rejection experiences among Ireland. Br J Psychiatry. 1997; 170: 447–52. PubMed Abstract | Publisher Full Text Harmon-Jones C, Bastian B, Harmon-Jones E: The Discrete Emotions
Questionnaire: A New Tool for Measuring State Self-Reported Emotions. PLoS One. 2016; 11(8): e0159915. |
Mallon DS, Galway DK: Towards an Understanding of the Role of
Bereavement in the Pathway to Suicide. KEES-suicide-bereavement-mallon-
and-galway. 2015; (2015–15). References ;
( )
PubMed Abstract | Publisher Full Text | Free Full Text Harwood D, Hawton K, Hope T, et al.: Psychiatric disorder and personality
factors associated with suicide in older people: a descriptive and case-
control study. Int J Geriatr Psychiatry. 2001; 16(2): 155–165. PubMed Abstract | Publisher Full Text Malone KM: Suicide in Ireland Survey. 2013. Malone KM, McGuinness SG, Cleary E, et al.: Lived Lives: A Pavee Perspective. An arts-science community intervention around suicide in an indigenous
ethnic minority [version 1; peer review: 3 approved]. Wellcome Open Res. 2017; 2: 27. Hasin DS, Sarvet AL, Meyers JL, et al.: Epidemiology of adult DSM-5 major
depressive disorder and its specifiers in the United States. JAMA psychiatry. 2018; 75(4): 336–346. Mann JJ, Apter A, Bertolote J, et al.: Suicide prevention strategies: a
systematic review. JAMA. 2005; 294(16): 2064–2074. PubMed Abstract | Publisher Full Text Henderson C, Noblett J, Parke H, et al.: Mental health-related stigma in
health care and mental health-care settings. Lancet Psychiatry. 2014; 1(6):
467–82. McGuinness S, Malone KM: Lived Lives: From Tory Island to Swift’s Asylum
E-publication. 2019. Henriksson MM, Aro HM, Marttunen MJ, et al.: Mental disorders and
comorbidity in suicide. Am J Psychiatry. 1993; 150(6): 935–940. Henriksson MM, Aro HM, Marttunen MJ, et al.: Mental disorders and
comorbidity in suicide. Am J Psychiatry. 1993; 150(6): 935–940. Open Peer Review Current Peer Review Status: Reviewer Report 04 April 2022 https://doi.org/10.21956/wellcomeopenres.19725.r49557 © 2022 Shidhaye R. This is an open access peer review report distributed under the terms of the Creative
Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium,
provided the original work is properly cited. Rahul Shidhaye Pravara Institute of Medical Sciences, Pune, Maharashtra, India I had a chance to look at the revised manuscript and I am satisfied with all the respo Competing Interests: No competing interests were disclosed. Reviewer Expertise: Global Mental Health, Yoga and Health, Implementation Science I confirm that I have read this submission and believe that I have an appropriate level of
expertise to confirm that it is of an acceptable scientific standard. Version 2 Reviewer Report 09 March 2022 https://doi.org/10.21956/wellcomeopenres.19095.r47950 © 2022 Shidhaye R. This is an open access peer review report distributed under the terms of the Creative
Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium,
provided the original work is properly cited. Publisher Full Text Lived Lives: Lived Lives I : A Rural Perspective; Lived Lives II: A Psychiatric
Hospital Perspective, 2020. [dataset]. Version 1. Irish Social Science Data
Archive. 2021. Ungar T, Knaak S, Szeto ACH: Theoretical and Practical Considerations for
Combating Mental Illness Stigma in Health Care. Community Ment Health J. 2016; 52(3): 262–271. Ungar T, Knaak S, Szeto ACH: Theoretical and Practical Considerations for
Combating Mental Illness Stigma in Health Care. Community Ment Health J. 2016; 52(3): 262–271. Lundberg B, Hansson L, Wentz E, et al.: Sociodemographic and clinical factors
related to devaluation/discrimination and rejection experiences among ( )
PubMed Abstract | Publisher Full Text | Free Full Text Page 15 of 23 Page 15 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Rahul Shidhaye Pravara Institute of Medical Sciences, Pune, Maharashtra, India Malone and colleagues present mixed-methods evaluation of 'Lived Lives' project in this extremely
well-written paper. I read this paper sitting in my office in a small village in rural Maharashtra
(western part of India), yet I felt that I was part of the various exhibits in St. Patrick's University
Hospital. I agree with the feedback from the participants that this was very emotional, moving, Page 16 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 and extremely powerful. I indeed felt sad, but glad and as I was reading the paper, I started
reflecting on what I could do as a psychiatrist to address the issues related to suicide,
bereavement, and stigma. I reviewed the revised version of the paper. Dr. Quinlivan had made
some excellent comments on the methodology and other aspects of the paper. Authors have
revised the paper based on these suggestions and Dr. Quinlivan seems to be satisfied with these
changes. As a result, I do not have any major comments on this version except for a few which
follow: and extremely powerful. I indeed felt sad, but glad and as I was reading the paper, I started
reflecting on what I could do as a psychiatrist to address the issues related to suicide,
bereavement, and stigma. I reviewed the revised version of the paper. Dr. Quinlivan had made
some excellent comments on the methodology and other aspects of the paper. Authors have
revised the paper based on these suggestions and Dr. Quinlivan seems to be satisfied with these
changes. As a result, I do not have any major comments on this version except for a few which
follow: and extremely powerful. I indeed felt sad, but glad and as I was reading the paper, I started
reflecting on what I could do as a psychiatrist to address the issues related to suicide,
bereavement, and stigma. I reviewed the revised version of the paper. Dr. Quinlivan had made
some excellent comments on the methodology and other aspects of the paper. Authors have
revised the paper based on these suggestions and Dr. Quinlivan seems to be satisfied with these
changes. Rahul Shidhaye It will be good if authors can elaborate on this in the discussion section,
particularly highlighting the limitations of the current evaluation plan. 5. Rahul Shidhaye As a result, I do not have any major comments on this version except for a few which
follow: It will be good to have clearly stated objectives and rationale for this particular paper
described at the end of the introduction section. On page 6, authors have included a
paragraph about 'Lived Lives II' and in this same paragraph, some of these details can be
included. 1. Similarly, an overview of the evaluation plan can be presented in the methods section
before going into the details of each of the methods. Importantly, explain why a particular
method was chosen and how it relates to other methods in this study. 2. Making stigma visible is a stated objective of this paper. It will be good if you can present
any data (qualitative) related to this theme. I found most of the things related to this as
some kind of statements from authors, but not entirely supported by the qualitative data. Maybe I am missing something here. 3. Mean and standard deviation for each of the sub-scales of Discrete Choice Questionnaire is
presented. For lay readers (like me), it will be good to explain these scores in context. 4. Mean and standard deviation for each of the sub-scales of Discrete Choice Questionnaire is
presented. For lay readers (like me), it will be good to explain these scores in context. 4. I agree that 'Lived Lives' is a potential community intervention to address stigma, suicide,
and bereavement issues. But the results presented in this paper only suggest that the
exhibit and guided tour is acceptable to those who attended. What was the effect of this on
their internalized/externalized stigma, emotional state, and long-term behavior is not
presented here. Additionally, it is also important to explore ways to engage with wider
community. It will be good if authors can elaborate on this in the discussion section,
particularly highlighting the limitations of the current evaluation plan. 5. I agree that 'Lived Lives' is a potential community intervention to address stigma, suicide,
and bereavement issues. But the results presented in this paper only suggest that the
exhibit and guided tour is acceptable to those who attended. What was the effect of this on
their internalized/externalized stigma, emotional state, and long-term behavior is not
presented here. Additionally, it is also important to explore ways to engage with wider
community. If applicable, is the statistical analysis and its interpretation appropriate?
Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Page 17 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Yes
Competing Interests: No competing interests were disclosed. Reviewer Expertise: Global Mental Health, Yoga and Health, Implementation Science
I confirm that I have read this submission and believe that I have an appropriate level of
expertise to confirm that it is of an acceptable scientific standard, however I have
significant reservations, as outlined above. Competing Interests: No competing interests were disclosed. I confirm that I have read this submission and believe that I have an appropriate level of
expertise to confirm that it is of an acceptable scientific standard, however I have
significant reservations, as outlined above. Author Response 24 Mar 2022 Dear Dr. Shidhaye, Thank you for taking the time to review this paper and for your generous feedback. Based on your comments, we have made the following changes:
We have added a clear objective at the end of the introduction section to orientate the
reader. We have also added an overview of the evaluation plan earlier in the methods
section as suggested and elaborated on the nature of the methodology for this and why it
was chosen. We have also added in explanations for the terms "mean" and "standard
deviation" in the results section and elaborated on the limitations of the current research in
terms of its cross-sectional nature with regard to emotional state. We have further reflected
on the limitations with regards to engaging the wider community who chose not to attend
the exhibition and how we might find out more about their choice in this regard. In response to your comments on the theme of "making stigma visible" we would like to
clarify that this was an overall theme for the project in SPUH and for the changes made to
the 21g artwork. Thus, it refers to a metaphor for what we hoped would be achieved
through installing the exhibition in this space as opposed to a research objective to be
measured or a "theme" in the qualitative data sense. We hope that answers your query in
this regard. We would like to take this opportunity to thank you again for your constructive comments. Your input and indeed the input of all reviewers have helped us improve the quality and
structure of our paper as we continue to navigate the best ways to present this research
which doesn't conform to the traditional "intro, methods, results, discussion" format. Kind Regards,
Lived Lives Research Team Kind Regards, g
Lived Lives Research Team Competing Interests: No competing interests were disclosed. Reviewer Report 21 December 2021 Page 18 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Leah Quinlivan 1 NIHR Greater Manchester Patient Safety Translational Research Centre, The University of
Manchester, Manchester Academic Health Science Centre, Manchester, UK
2 Centre for Mental Health and Safety, University of Manchester, Manchester, UK 1 NIHR Greater Manchester Patient Safety Translational Research Centre, The University of
Manchester, Manchester Academic Health Science Centre, Manchester, UK
2 C
t
f
M
t l H
lth
d S f t
U i
it
f M
h
t
M
h
t
UK 1 NIHR Greater Manchester Patient Safety Translational Research Centre, The Unive
Manchester, Manchester Academic Health Science Centre, Manchester, UK 2 Centre for Mental Health and Safety, University of Manchester, Manchester, UK l queries are well addressed in the revised version. No further changes are requested This is excellent paper, well written, and emotive to read. To truly transform mental health
services at a human level and address societal stigma, we need these innovative and
groundbreaking ‘Lived Lives’ collaborations between art and science. I couldn't sum up the impact
of this work greater than the participants in the event: “there is something quite powerful about the fact that we are here – in this room, and statistically I
could be on the wall and I’m not, and I’m very grateful I’m not and that gives me encouragement
and hope”” (M2, person with lived experience) “out of a sense of failure there is a power in the vulnerability. It can be very empowering to
experience something like this and re-invigorate our passion to make a difference”. (F5,
Administration Director) “out of a sense of failure there is a power in the vulnerability. It can be very empowering to
experience something like this and re-invigorate our passion to make a difference”. (F5,
Administration Director) “Very moving and emotional. It brought the people who died by suicide to a living state and their
families feelings, so real. Thanks for this experience.” (P51) “Very moving and emotional. It brought the people who died by suicide to a living state and their
families feelings, so real. Thanks for this experience.” (P51) Best of luck to the team in their innovative endeavours. I look forward to seeing this intervention
develop its transformative reach further into society. https://doi.org/10.21956/wellcomeopenres.19095.r47711 © 2021 Quinlivan L. This is an open access peer review report distributed under the terms of the Creative
Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium,
provided the original work is properly cited. I confirm that I have read this submission and believe that I have an appropriate level of
expertise to confirm that it is of an acceptable scientific standard. I confirm that I have read this submission and believe that I have an appropriate level of
expertise to confirm that it is of an acceptable scientific standard. Version 1 Reviewer Report 23 June 2021 https://doi.org/10.21956/wellcomeopenres.17070.r44154 © 2021 Quinlivan L. This is an open access peer review report distributed under the terms of the Creative
Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium,
provided the original work is properly cited. Competing Interests: No competing interests were disclosed. Reviewer Expertise: Public engagement, sensitive research methods, suicide prevention, self-
harm, clinical research. Are sufficient details of methods and analysis provided to allow replication by others?
Yes Are the conclusions drawn adequately supported by the results? Page 19 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Competing Interests: No competing interests were disclosed. and could help reduce suicidal feelings. This is a ground-breaking and important programme of work which leads the way internationally
for co-production and public engagement in suicide prevention and mental health research. The
authors have taken highly stigmatised and often abstract concepts and placed them in the reality
of the devastating loss when someone dies by suicide. The publication is strong and interesting/
emotive to read. Some minor comments which the authors may which to address prior to
indexing. In the first section on depression and suicide in Ireland. The authors use the term ‘post-
attempter’. Can this be replaced by an alternative term such as ‘post suicide attempt’, to be
consistent with the ethos of the paper. 1. Some of the images are a blurry. Suspect this is due to uploading issues. Please do check
these prior to submitting a new version. The photographs are outstanding and an integral
part of the publication. 2. The term ‘show-back’ is used in various places throughout in the document. May be helpful
to define or re-word this term. I have no idea what it means. 3. Some restructure and tightening of the methods may be helpful to readers. 4. Some restructure and tightening of the methods may be helpful to readers. 4. In the abstract [method section]. May be helpful to be clearer about the methods. Something like ‘a mixed method approach was used to evaluate an arts-led intervention to
address stigma’. 5. The method section may benefit from a slight restructure, and additional use of headings
(e.g., ethical approval/ ethics approach, procedure(s), questionnaire, participants, analyses). 6. It would be helpful to see a more detailed analyses section. Why did the authors use
ANOVA? What is a ‘brief’ thematic analyses? 7. There are a few different numbers for participants in the text. A defined participant section
may help make this clearer. For example, in the abstract it states 86 people engaged with
the exhibition, but in the attendees there are 58 delegates, and later in the results 68
people completed the questionnaire. Appreciate there is different levels of engagement and
completion of the questionnaire, but can this be made clearer for the readers? 8. Quinlivan, L., Gorman, L., Littlewood, D. L., Monaghan, E., Barlow, S. J., Campbell, S., ... & Kapur, N.
(2021). ‘Wasn’t offered one, too poorly to ask for one’–Reasons why some patients do not receive a Leah Quinlivan 1 NIHR Greater Manchester Patient Safety Translational Research Centre, The University of
Manchester, Manchester Academic Health Science Centre, Manchester, UK
2 Centre for Mental Health and Safety, University of Manchester, Manchester, UK 1 NIHR Greater Manchester Patient Safety Translational Research Centre, The University of
Manchester, Manchester Academic Health Science Centre, Manchester, UK 2 Centre for Mental Health and Safety, University of Manchester, Manchester, UK This novel study evaluates the use of a psychobiographical and visual arts autopsy led
engagement event as an ‘intervention’ to address the stigma of suicide in the community. This
combination of art and science brings forward the identity of the person lost to suicide into the
public domain. This unique approach to public engagement involves researchers, clinicians,
artists, families, and communities to facilitate dialogue of highly stigmatised and sensitive topics,
in a safe but challenging space. Based on evidence from previous engagement events, the authors
situated the mediated exhibition within a psychiatric institution, where experiences of stigma are
persistently high. Hosting an event such as this in health services is of great significance. Despite progress in the
development of mental health services, patients frequently speak of receiving stigmatising
comments when they seek help after harming themselves (Quinlivan et al., 2021 a,b). Statistics
surrounding suicide can also create distance between professionals and the devastating human
experience when someone loses their life to suicide. Novel engagement approaches such as
these, are needed to bring forward the lived life of those lost to suicide compassionately, whilst
challenging the stigma surrounding suicidal behaviour. The authors conducted a mixed-methods evaluation of the exhibition which was attended by 86
participants. Emotional responses were intense but well managed with openly available
bereavement support. Most participants felt the engagement event could be helpful after suicide Page 20 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 References 1. Quinlivan L, Gorman L, Littlewood DL, Monaghan E, et al.: 'Wasn't offered one, too poorly to ask
for one' - Reasons why some patients do not receive a psychosocial assessment following self-
harm: Qualitative patient and carer survey.Aust N Z J Psychiatry. 2021. 48674211011262 PubMed
Abstract | Publisher Full Text 2. Quinlivan L, Gorman L, Littlewood D, Monaghan E, et al.: ‘Relieved to be seen’—patient and carer
experiences of psychosocial assessment in the emergency department following self-harm:
qualitative analysis of 102 free-text survey responses. BMJ Open. 2021; 11 (5). Publisher Full Text and could help reduce suicidal feelings. O
ll
hi i
ll
I f ll
i h h
h
f h
k “Lived Lives sits comfortably in discomfort, unafraid to call out the home-truths about stigma and
its pervasive and pernicious impact, managing to do so with deepest respect and empowerment,
and with restoring identity at its core.” References: Quinlivan, L., Gorman, L., Littlewood, D. L., Monaghan, E., Barlow, S. J., Campbell, S., ... & Kapur, N. (2021). ‘Wasn’t offered one, too poorly to ask for one’–Reasons why some patients do not receive a Page 21 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 psychosocial assessment following self-harm: Qualitative patient and carer survey. Australian &
New Zealand Journal of Psychiatry, 00048674211011262. https://doi.org/10.1177/00048674211011262 Quinlivan, L. M., Gorman, L., Littlewood, D. L., Monaghan, E., Barlow, S. J., Campbell, S. M., ... &
Kapur, N. (2021). ‘Relieved to be seen’—patient and carer experiences of psychosocial assessment
in the emergency department following self-harm: qualitative analysis of 102 free-text survey
responses. BMJ open, 11(5), e044434. http://dx.doi.org/10.1136/bmjopen-2020-044434 Are the conclusions drawn adequately supported by the results?
Yes I confirm that I have read this submission and believe that I have an appropriate level of
expertise to confirm that it is of an acceptable scientific standard, however I have
significant reservations, as outlined above. Page 22 of 23 Wellcome Open Research 2022, 6:85 Last updated: 04 APR 2022 Author Response 08 Dec 2021
kevin malone, University College Dublin, Dublin, Ireland
Dear Dr Quinlivan
Thank you for taking the time to review our paper and for your valuable insights and
comments. Based on your feedback, we have changed the term "post-attempter" to "post suicide
attempt" and acknowledge the original language used was an oversight and not in line with
the ethos of our paper nor our research. We have also added a definition of the term
"show-back", which we hope will clarify this important part of the research process for
readers. The novel nature of this arts-science collaboration and its methods can present a challenge
in terms of structuring papers in the traditional "introduction, methods, results, discussion,
format". We have however, based on your feedback, attempted to clarify the methodology
used in the abstract and also added some additional information specific to analyses used
with the questionnaire data collected. Furthermore, we have clarified participant numbers
throughout the paper and thank you for bringing this important point to our attention as
we can see it may have been confusing for readers to keep track of the different types of
"participants" we had. We hope you find these changes have improved the quality of our paper and we thank you
again for your input. Kind Regards,
Lived Lives Research Team
Competing Interests: No competing interests were disclosed. Author Response 08 Dec 2021
kevin malone, University College Dublin, Dublin, Ireland
Dear Dr Quinlivan Author Response 08 Dec 2021
kevin malone, University College Dublin, Dublin, Ireland Thank you for taking the time to review our paper and for your valuable insights an
comments. Based on your feedback, we have changed the term "post-attempter" to "post suicide
attempt" and acknowledge the original language used was an oversight and not in line with
the ethos of our paper nor our research. We have also added a definition of the term
"show-back", which we hope will clarify this important part of the research process for
readers. Are the conclusions drawn adequately supported by the results?
Yes The novel nature of this arts-science collaboration and its methods can present a challenge
in terms of structuring papers in the traditional "introduction, methods, results, discussion,
format". We have however, based on your feedback, attempted to clarify the methodology
used in the abstract and also added some additional information specific to analyses used
with the questionnaire data collected. Furthermore, we have clarified participant numbers
throughout the paper and thank you for bringing this important point to our attention as
we can see it may have been confusing for readers to keep track of the different types of
"participants" we had. We hope you find these changes have improved the quality of our paper and we thank you
again for your input. Kind Regards, Kind Regards,
Lived Lives Research Team g
Lived Lives Research Team Competing Interests: No competing interests were disclosed. Page 23 of 23
|
https://openalex.org/W3165364497
|
https://www.mdpi.com/2072-6643/13/6/1873/pdf?version=1622538672
|
English
| null |
Impact of Body Mass Index on the Survival of Patients with Sepsis with Different Modified NUTRIC Scores
|
Nutrients
| 2,021
|
cc-by
| 13,630
|
Citation: Tsai, Y.-H.; Lin, C.-Y.;
Chen, Y.-M.; Chang, Y.-P.; Hung, K.-Y.;
Chang, Y.-C.; Chen, H.-C.;
Huang, K.-T.; Chen, Y.-C.;
Wang, Y.-H.; et al. Impact of Body
Mass Index on the Survival of
Patients with Sepsis with Different
Modified NUTRIC Scores. Nutrients
2021, 13, 1873. https://doi.org/
10.3390/nu13061873 Abstract: Nutritional status affects the survival of patients with sepsis. This retrospective study
analyzed the impact of body mass index (BMI) and modified nutrition risk in critically ill (mNUTRIC)
scores on survival of these patients. Data of 1291 patients with sepsis admitted to the intensive care
unit (ICU) were extracted. The outcomes were mortality, duration of stay, ICU stay, and survival
curve for 90-day mortality. Logistic regression analysis was performed to examine the risk factors
for mortality. Cytokine and biomarker levels were analyzed in 165 patients. The 90-day survival of
underweight patients with low mNUTRIC scores was significantly better than that of normal-weight
patients with low mNUTRIC scores (70.8% vs. 58.3%, respectively; p = 0.048). Regression model
analysis revealed that underweight patients with low mNUTRIC scores had a lower risk of mortality
(odds ratio = 0.557; p = 0.082). Moreover, normal-weight patients with low mNUTRIC scores had the
lowest human leukocyte antigen DR (HLA-DR) level on days 1 (underweight vs. normal weight vs. overweight: 94.3 vs. 82.1 vs. 94.3, respectively; p = 0.007) and 3 (91.8 vs. 91.0 vs. 93.2, respectively;
p = 0.047). Thus, being underweight may not always be harmful if patients have optimal clinical
nutritional status. Additionally, HLA-DR levels were the lowest in patients with low survival. Received: 13 May 2021
Accepted: 26 May 2021
Published: 30 May 2021 Keywords: sepsis; nutrition; survival; underweight Publisher’s Note: MDPI stays neutral
with regard to jurisdictional claims in
published maps and institutional affil-
iations. Article
Impact of Body Mass Index on the Survival of Patients with
Sepsis with Different Modified NUTRIC Scores Yi-Hsuan Tsai 1, Chiung-Yu Lin 1, Yu-Mu Chen 1,2, Yu-Ping Chang 1
, Kai-Yin Hung 1,3, Ya-Chun Chang 1,
Hung-Cheng Chen 1,2
, Kuo-Tung Huang 1,2, Yung-Che Chen 1
, Yi-Hsi Wang 1,4, Chin-Chou Wang 1,4,5,
Meng-Chih Lin 1,4 and Wen-Feng Fang 1,4,5,* 1
Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine,
Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine,
Kaohsiung 83301, Taiwan; yhtsai@cgmh.org.tw (Y.-H.T.); chiungyu@cgmh.org.tw (C.-Y.L.);
blackie@cgmh.org.tw (Y.-M.C.); b9002087@cgmh.org.tw (Y.-P.C.); redrosahung@yahoo.com.tw (K.-Y.H.);
y7817@cgmh.org.tw (Y.-C.C.); chc1106@cgmh.org.tw (H.-C.C.); jelly@cgmh.org.tw (K.-T.H.);
yungchechen@yahoo.com.tw (Y.-C.C.); yihsi@cgmh.org.tw (Y.-H.W.); ccwang5202@yahoo.com.tw (C.-C.W.);
mengchih@cgmh.org.tw (M.-C.L.) g
g
g
2
Graduate Institute of Clinical Medical Sciences, Chang Gung University, Taoyuan 33302, Taiwan
3
D
t
t f N t iti
l Th
K
h i
Ch
G
M
i l H
it l K
h i
83301 T i 2
Graduate Institute of Clinical Medical Sciences, Chang Gung University, Taoyuan 33302, Taiwan
3
Department of Nutritional Therapy, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung 83301, Taiwan
4
Department of Respiratory Therapy, Kaohsiung Chang Gung Memorial Hospital,
Ch
G
U i
it
C ll
f M di i
K
h i
83301 T i g
g
y
g
g
5
Department of Respiratory Care, Chang Gung University of Science and Technology, Chiayi 61363, Taiwan
*
Correspondence: fangwf@hotmail.com; Tel.: +886-7-7317123 (ext. 8199) 5
Department of Respiratory Care, Chang Gung University of Science and Technology, Chiayi 61363, Taiwan
*
Correspondence: fangwf@hotmail com; Tel : +886 7 7317123 (ext 8199)
Citation: Tsai, Y.-H.; Lin, C.-Y.;
Chen, Y.-M.; Chang, Y.-P.; Hung, K.-Y.;
Chang, Y.-C.; Chen, H.-C.;
Huang, K.-T.; Chen, Y.-C.;
Wang, Y.-H.; et al. Impact of Body
Mass Index on the Survival of
Patients with Sepsis with Different
Modified NUTRIC Scores. Nutrients
2021, 13, 1873. https://doi.org/
10.3390/nu13061873
Academic Editor: Matthew
C. Lohman
Received: 13 May 2021
Accepted: 26 May 2021
Published: 30 May 2021
Citation: Tsai, Y.-H.; Lin, C.-Y.;
Chen, Y.-M.; Chang, Y.-P.; Hung, K.-Y.;
Chang, Y.-C.; Chen, H.-C.;
Huang, K.-T.; Chen, Y.-C.;
Wang, Y.-H.; et al. Impact of Body
Mass Index on the Survival of
Patients with Sepsis with Different
Modified NUTRIC Scores. Nutrients
2021, 13, 1873. https://doi.org/
10.3390/nu13061873
Academic Editor: Matthew
C. Lohman
Received: 13 May 2021
Accepted: 26 May 2021
Published: 30 May 2021 nutrients nutrients nutrients nutrients 1. Introduction Sepsis is characterized by life-threatening organ dysfunction caused by dysregulated
host response to infection [1]. According to an Asian transnational study, the in-hospital
mortality rate of patients with sepsis was reported to be 44.5% [2]. Previous studies
have identified risk factors for mortality, such as comorbidities [3,4], poor nutrition [5,6],
abnormal clinical parameters [7,8], and impaired immunity [3,9,10], in patients with sepsis. Identification of simple phenotypes in patients with strong survivability is difficult because
sepsis is highly heterogeneous [11]. Nevertheless, identification of phenotypes or sub-
phenotypes of patients with sepsis with different survival rates would be promising for
clinical risk stratification. Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article
distributed
under
the
terms
and
conditions of the Creative Commons
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
4.0/). Nutritional status is likely to be a phenotype of sepsis [12,13]. Previous studies have
reported that half of hospital in-patients have nutritional problems [14–16]. Malnutri- https://www.mdpi.com/journal/nutrients Nutrients 2021, 13, 1873. https://doi.org/10.3390/nu13061873 Nutrients 2021, 13, 1873 2 of 17 tion in patients with sepsis may increase the risk of mortality [17–24] and duration of
ventilator dependence [25]. In these studies, body mass index (BMI), a simple variable
calculated using the following formula: BMI = kg/m2 (kg: a person’s weight in kilograms;
m2: person’s height in meters squared), was commonly used to indicate the nutritional sta-
tus of the general population. However, the association between BMI and patient outcomes
remains controversial. Overweight and obesity may be associated with lower mortality than
normal weight [23,26–32], owing to the higher metabolic reserves in acute catabolic illnesses
among individuals with higher BMIs. In addition, adipose tissue secretes anti-inflammatory
mediators, such as leptin and tumor necrosis factor (TNF) receptor 2 [33]. In contrast, some
studies reported that overweight and obesity increased the risk of mortality [20–22,34,35],
especially in patients with cardiovascular disease [20,35], diabetes, and kidney disease [20]. Other studies have shown no significant differences in the risk of mortality between obese
and normal-weight critically ill patients [36–39]. These controversial results imply that true
nutritional status cannot be represented by simple BMI-related stratifications. Therefore, scores have been developed to evaluate the nutritional status of critically ill
patients [6,12,13]. The modified nutrition risk in critically ill (mNUTRIC) score simplifies
the NUTRIC score by excluding the interleukin 6 (IL-6) variable. 1. Introduction It includes age, the
acute physiology and chronic health evaluation II (APACHE II) score, the sequential organ
failure assessment (SOFA) score, number of comorbidities, and duration of admission to
the intensive care unit (ICU). This score has been verified for the nutritional evaluation
of critically ill patients. A study showed that patients with higher mNUTRIC scores had
significantly higher mortality rates [6]. However, the mNUTRIC score does not include the
BMI domain and is more complex than the BMI. In addition, sepsis-induced cytokine changes related immune dysfunction have an
impact on clinical outcomes [9]. Fajgenbaum and June’s review revealed that IL-6, IL-1,
IL-17, TNF, and interferon gamma (IFN-γ) were released during a cytokine storm. IL-10
and IL-1 receptor antagonist (IL-1RA) are negative regulators of cytokine storms [40]. In our
previous studies, lower human leukocyte antigen DR (HLA-DR) expression was associated
with higher mortality due to nutritional insufficiency [5]. Patients had a higher risk of
mortality when IL-6 and TNF-α levels were higher [9]. However, it remains unclear how
cytokine levels differ among patients with sepsis who have different nutritional statuses. Because nutritional status affects the outcomes of patients with sepsis, it is important to
identify the role of cytokines in nutrition and sepsis. y
y
p
In this study, we proposed a nutritional phenotype based on complete nutritional
status that consists of BMI and the mNUTRIC score. To examine how this nutritional
phenotype affects the survival of patients with sepsis, this study analyzed the impact of
BMI and the mNUTRIC score on the survival of these patients. Based on the conceptual
framework mentioned above, this study aimed to determine the following: (a) survival of
patients with sepsis classified into different BMI and mNUTRIC score groups, (b) impact
of the mNUTRIC scores on the survival of patients with sepsis who had different BMIs,
(c) impact of the BMI on the survival of patients with sepsis who had different mNUTRIC
scores, and (d) the cytokine levels linked with different nutritional phenotypes. Specifically,
we used multiple domains and steps to identify the heterogeneity of nutritional status and
clarify its impact on the survival of patients with sepsis. Biological data analyses were
performed to confirm the results. Our results may help in further understanding whether
the nutritional phenotype influences survival of patients with sepsis, which could help
improve their care and treatment. 2.2. Definitions This study used the mNUTRIC score and BMI to evaluate the nutritional status of
the participants. The NUTRIC score includes six parameters: age, APACHE II score,
SOFA score, number of comorbidities, IL-6 level, and hospital stay before admission to the
ICU [12]. The mNUTRIC score includes all these parameters except IL-6 [6] (Table 1). In
our cohort, the median mNUTRIC score was 6, mode was 6, and mean was 6.01. Thus, we
chose 6 as the cut-off value for classification into low and high mNUTRIC score groups. Participants with mNUTRIC scores of ≥6 were classified into the high mNUTRIC score
group, and those with mNUTRIC scores of <6 were included in the low mNUTRIC score
group. BMI was defined as the value obtained by dividing the weight of the participant in
kilograms by the square of the height in meters. This was calculated when the patients were
admitted to the ICU. BMI was categorized into three groups: underweight (<18.5 kg/m2),
normal-weight (18.5–24.9 kg/m2), and overweight (≥25.0 kg/m2). Table 1. modified NUTRIC score. Variable
Range
Points
Age
<50
0
≥50 and <75
1
≥75
2
APACHE II
<15
0
≥15 and <20
1
≥20 and <28
2
≥28
3
SOFA
<6
0
≥6 and <10
1
≥10
2
Number of comorbidities
0–1
0
≥2
1
Days from hospital to ICU admission
≥0 and <1
0
≥1
1
APACHE II: acute physiology and chronic health evaluation II; SOFA: sequential organ failure assessment; ICU:
Intensive Care Unit. Table 1. modified NUTRIC score. 2. Materials and Methods
2.1. Study Participants This retrospective study enrolled adult patients with sepsis who were admitted to
medical ICUs (34 beds) at Kaohsiung Chang Gung Memorial Hospital between August
2013 and January 2017. Patients with sepsis who were admitted to the medical ICU between
January 2020 and August 2020 were included in the validation cohort. All participants Nutrients 2021, 13, 1873 3 of 17 3 of 17 who fulfilled the criteria of the Third International Consensus Definitions for Sepsis and
Septic Shock (Sepsis-3) [1] were screened. Clinical parameters, including age, sex, BMI,
APACHE II score, Charlson comorbidity index, SOFA score, NUTRIC score, comorbidities,
infection site, laboratory data, and day of mortality, were obtained from medical records. In addition, the study enrolled 165 patients who agreed to undergo blood sampling during
ICU hospitalization. All patients or family members provided written informed consent
for the use of residual blood samples. The study design was approved by the Institutional
Review Board (ID: 202001696B0C501) of Chang Gung Memorial Hospital. 2.3. Biomarkers The levels of HLA-DR monocyte expression and cytokines, including IL-6, IL-1RA,
IL-10, IL-17, TNF-α, and IFN-γ in the plasma, have been described in our previous re-
ports [3,5,7,9]. All data were collected after admission to the ICU on days 1, 3, and 7. 2.4. Outcome The main outcome of this study was survival rate. We compared 7-day, 14-day, 28-day,
and 90-day mortality rates between patients in the different BMI and mNUTRIC score
groups. The 90-day mortality rate was used to construct the survival curves. The secondary
outcomes were length of stay in the hospital and ICU. Nutrients 2021, 13, 1873 4 of 17 2.5. Statistical Analyses 2.5. Statistical Analyses The data analysis was performed using SPSS software (version 22.0; IBM Corp.,
Armonk, NY, USA). The high and low mNUTRIC scores were compared in terms of
demographic characteristics, baseline characteristics, and outcomes. The Mann-Whitney
U test was used for continuous variables and chi-square test was used for categorical
variables. The survival curve was constructed according to the 90-day mortality that was
determined based on the mNUTRIC score. Subsequently, the demographic characteristics,
baseline characteristics, and outcomes were compared across the underweight, normal-
weight, and overweight groups. The Kruskal-Wallis test was used for continuous variables
and chi-square test was used for categorical variables. The survival curve was constructed
according to the 90-day mortality that was determined based on the different BMI groups. For the three BMI groups, we created a survival curve from the 90-day mortality based on
the mNUTRIC scores. In the low and high mNUTRIC score subgroups, we compared between underweight,
normal-weight, and overweight individuals. The survival curve was constructed according
to the 90-day mortality that was determined based on the different BMI groups. To identify
the variables that were associated with mortality, we constructed a logistic regression
model to analyze the variables with significant differences using the Kruskal-Wallis and
chi-square tests. We analyzed the validation cohort to confirm the results. q
y
The dependent variable of the logistic regression model was the 90-day mortality
rate. The unadjusted model (Model 1) included intercept, age, sex, and different BMI
groups (baseline: normal-weight). Models 2–5 were adjusted for the same variables used
in Model 1. Model 2 further included the SOFA score on day 1, Model 3 further included
comorbidities, Model 4 further included infection sites, and Model 5 further included
glucose parameters. Model 6 included all the variables used in Models 1–5. Finally, we analyzed the cytokine and biomarker data of 165 participants to investi-
gate the biological mechanisms of sepsis-related mortality. We analyzed the biomarker
differences across the three BMI groups in the low and high mNUTRIC score subgroups. The biomarkers that were analyzed included IL-6, IL-1RA, IL-10, IL-17, TNF-α, IFN-γ, and
HLA-DR after ICU admission on days 1, 3, and 7. 3.1. Baseline Characteristics of Patients with Low and High mNUTRIC Scores The construction cohort of this study included 513 patients with sepsis who had high
mNUTRIC scores and 285 patients with low mNUTRIC scores. Table 2 shows the baseline
characteristics and outcomes of the study participants. The validation cohort of the study
included 245 patients with sepsis who had high mNUTRIC scores and 245 patients with
low mNUTRIC scores. Supplementary Table S1 shows the baseline characteristics and
outcomes of the study participants in the validation cohort. In the construction cohort, the mean age of the participants was 67.1 years, and the
patients with high mNUTRIC scores were older than those with low mNUTRIC scores
(71.1 years vs. 60.0 years, respectively; p < 0.001). The mean ages were comparable in
the validation cohort. Patients with sepsis who had high mNUTRIC scores had higher
mean APACHE II scores (28.5 vs. 18.1; p < 0.001), Charlson comorbidity index (mean of 2.8
vs. 2.3; p < 0.001), and mean SOFA scores (10.2 vs. 6.8; p < 0.001). The APACHE II score,
Charlson comorbidity index, and SOFA score were comparable in the validation cohort. Patients with sepsis who had high mNUTRIC scores also had a higher mean number of
comorbidities (2.0 vs. 1.1; p < 0.001), such as coronary artery disease (32.0% vs. 14.0%;
p < 0.001), stroke (23.2% vs. 9.8%; p < 0.001), hypertension (62.0% vs. 44.4%, p < 0.001),
CKD (38.4% vs. 14.7%; p < 0.001), and diabetes mellitus (52.8% vs. 29.8%; p < 0.001),
than those with low mNUTRIC scores. In the validation cohort, patients with sepsis who
had high mNUTRIC scores also had a higher number of comorbidities; in addition, the
prevalence values of stroke, hypertension, chronic kidney disease (CKD), and diabetes
mellitus were comparable. 5 of 17 Nutrients 2021, 13, 1873 Table 2. Baseline characteristics and outcomes of the patients with sepsis in the construction cohort of the study. 3.1. Baseline Characteristics of Patients with Low and High mNUTRIC Scores eline characteristics and outcomes of the patients with sepsis in the construction cohort of the study. *** p < 0.001; ** p < 0.01; * p < 0.05; † p < 0.09; abbreviations: SD, standard deviation; mNUTRIC, modified nutrition risk in critically ill; BMI,
body mass index; APACHE, acute physiology and chronic health evaluation; SOFA, sequential organ failure assessment; COPD, chronic
obstructive pulmonary disease; CKD, chronic kidney disease; UTI, urinary tract infection; HbA1c, hemoglobin A1c; ICU, intensive care
unit; LOS, length of stay. When comparing the high mNUTRIC and low mNUTRIC score groups, the propor-
tions of patients with pneumonia (61.6% vs. 69.5%; p = 0.026) and soft tissue infection (3.5%
vs. 7.9%; p = 0.009) were lower in the former than in the latter. First-day capillary glucose
levels in patients admitted to the ICU were greater in the high mNUTRIC score group than
in the low mNUTRIC score group (213.8% vs. 189.5%, respectively; p < 0.001). Moreover,
the 7-day, 14-day, 28-day, and 90-day mortality rates were significantly lower in the low
mNUTRIC score group than in the high mNUTRIC score group (7-day mortality rate: 16.2%
vs. 6.3%, respectively; p < 0.001) (14-day mortality rate: 22.8% vs. 11.9%, respectively;
p < 0.001) (28-day mortality rate: 32.0% vs. 21.8%, respectively; p < 0.001) (90-day mortality
rate: 46.8% vs. 34.4%, respectively; p < 0.001). 3.1. Baseline Characteristics of Patients with Low and High mNUTRIC Scores mNUTRIC Score on
the First Day
All
(n = 799)
mNUTRIC
Score ≥6
(n = 513)
mNUTRIC
Score ≤5
(n = 286)
p
Under-Weight
(n = 149)
Normal-
Weight
(n = 405)
Overweight
(n = 245)
p
Demographic characteristics, mean ± SD
Age (years)
67.1 ± 14.9
71.1 ± 12.5
60.0 ± 16.1
<0.001 ***
68.5 ± 15.2
68.0 ± 14.4
64.7 ± 15.2
0.013 *
BMI, kg/m2
22.8 ± 4.9
22.9 ± 4.7
22.7 ± 5.2
0.464
16.5 ± 1.5
21.7 ± 1.8
28.8 ± 3.5
<0.001 ***
Sex (female), n (%)
323 (40.5)
215 (41.9)
108 (37.9)
0.268
45 (30.2)
154 (38.0)
124 (50.6)
<0.001 ***
Score, mean ± SD
APACHE II
24.7 ± 8.4
28.5 ± 6.9
18.1 ± 6.4
<0.001 ***
23.8 ± 7.5
25.3 ± 8.3
24.4 ± 9.0
0.094
Charlson comorbidity
index
2.6 ± 2.0
2.8 ± 1.9
2.3 ± 2.1
<0.001 ***
2.7 ± 2.1
2.7 ± 2.0
2.4 ± 1.9
0.315
SOFA
9.0 ± 3.8
10.2 ± 3.4
6.8 ± 3.5
<0.001 ***
8.2 ± 3.5
8.9 ± 3.7
9.7 ± 4.1
0.001 **
mNUTRIC
6.0 ± 1.8
7.1 ± 1.0
4.1 ± 1.1
<0.001 ***
5.8 ± 1.8
6.1 ± 1.7
5.9 ± 1.8
0.127
Comorbidities (number)
1.7 ± 1.2
2.0 ± 1.1
1.1 ± 1.0
<0.001 ***
1.5 ± 1.2
1.8 ± 1.2
1.6 ± 1.2
0.014 *
Comorbidities, n (%)
Coronary artery disease
204 (25.5)
164 (32.0)
40 (14.0)
<0.001 ***
26 (17.4)
117 (28.9)
61 (24.9)
0.023 *
History of stroke
147 (18.4)
119 (23.2)
28 (9.8)
<0.001 ***
35 (23.5)
79 (19.5)
33 (13.5)
0.032 *
Hypertension
444 (55.6)
318 (62.0)
126 (44.4)
<0.001 ***
73 (49.0)
225 (55.7)
146 (59.6)
0.121
COPD
117 (14.6)
82 (16.0)
34 (11.9)
0.119
24 (16.1)
72 (17.8)
21 (8.6)
0.005 **
Cancer
190 (23.8)
127 (25.0)
62 (21.8)
0.309
39 (26.4)
112 (28.0)
39 (16.0)
0.002 **
CKD
239 (29.9)
197 (38.4)
42 (14.7)
<0.001 ***
32 (21.5)
125 (30.9)
82 (33.5)
0.035 *
Liver cirrhosis
69 (8.6)
50 (9.7)
19 (6.7)
0.138
9 (6.0)
33 (8.1)
27 (11.0)
0.206
Diabetes mellitus
356 (44.6)
271 (52.8)
85 (29.8)
<0.001 ***
48 (32.1)
176 (43.5)
132 (53.9)
<0.001 ***
Site of suspected infection, n (%)
Lung
515 (64.5)
316 (61.6)
198 (69.5)
0.026 *
119 (79.9)
271 (66.9)
125 (51.0)
<0.001 ***
UTI
169 (21.2)
113 (22.0)
56 (19.6)
0.431
13 (22.1)
79 (19.5)
57 (23.3)
0.496
Intraabdominal
infection
57 (7.1)
40 (7.8)
17 (6.0)
0.336
5 (3.4)
33 (8.1)
19 (7.8)
0.137
Soft tissue infection
40 (5.0)
18 (3.5)
22 (7.7)
0.009 **
8 (5.4)
14 (3.5)
18 (7.3)
0.086 †
Bacteraemia
61 (7.6)
44 (8.6)
17 (6.0)
0.183
6 (4.0)
27 (6.7)
28 (11.4)
0.016 *
Baseline glucose and HbA1c, mean ± SD
HbA1c (%) **
7.3 ± 2.2
7.3 ± 2.3
7.2 ± 2.1
0.453
6.9 ± 2.0
7.1 ± 2.1
7.6 ± 2.4
0.014 *
Glucose (mg/dL)
205.7 ± 117.6
213.8 ± 106.2
189.5 ± 136.7
<0.001 ***
172.1 ± 73.7
210.7 ± 135.9
216.9 ± 102.2
<0.001 ***
Admission days, mean ± SD
ICU days
12.5 ± 9.7
12.3 ± 9.5
12.9 ± 10.1
0.411
14.0 ± 10.0
12.0 ± 9.2
12.3 ± 10.3
0.035 *
LOS
30.1 ± 28.4
30.5 ± 31.0
29.5 ± 23.0
0.364
35.6 ± 32.3
28.6 ± 28.1
29.3 ± 26.1
0.070 †
Mortality, n (%)
7-day mortality
102 (12.8)
83 (16.2)
18 (6.3)
<0.001 ***
13 (8.7)
54 (13.3)
35 (14.3)
0.245
14-day mortality
152 (19)
117 (22.8)
34 (11.9)
<0.001 ***
23 (15.4)
81 (20.0)
48 (19.6)
0.461
28-day mortality
227 (28.4)
164 (32.0)
62 (21.8)
0.002 **
35 (23.5)
120 (29.6)
72 (29.4)
0.335
90-day mortality
339 (42.4)
240 (46.8)
98 (34.4)
0.001 **
60 (40.3)
181 (44.7)
98 (40.0)
0.422
*** p < 0.001; ** p < 0.01; * p < 0.05; † p < 0.09; abbreviations: SD, standard deviation; mNUTRIC, modified nutrition risk in critically ill; BMI,
body mass index; APACHE, acute physiology and chronic health evaluation; SOFA, sequential organ failure assessment; COPD, chronic
obstructive pulmonary disease; CKD, chronic kidney disease; UTI, urinary tract infection; HbA1c, hemoglobin A1c; ICU, intensive care
unit; LOS, length of stay. 3.2. Baseline Characteristics of Patients in the Different BMI Groups The construction cohort of this study was divided into three groups: underweight
(n = 149), normal-weight (n = 405), and overweight (n = 245) (Table 2). The validation
cohort of the study was also divided into three groups according to BMI: underweight
(n = 81), normal-weight (n = 241), and overweight (n = 170) (Table S1). g
g
In
the
construction
cohort,
the
overweight
group
was
the
youngest
(mean age = 64.7 years; p = 0.013), had the highest proportion of females (50.6%;
p < 0.001), and had the highest mean SOFA score (9.7; p < 0.001) (the SOFA scores were
comparable in the validation cohort). This group had the lowest proportions of patients Nutrients 2021, 13, 1873 6 of 17 with stroke (13.5%; p < 0.032), chronic obstructive pulmonary disease (COPD) (8.6%;
p = 0.005), and cancer (16.0%; p = 0.002); highest proportion of patients with CKD (33.5%;
p = 0.035), diabetes mellitus (53.9%; p < 0.001), bacteremia (11.4%; p = 0.016); and highest
mean HbA1c (7.6%; p = 0.014), and capillary glucose (216.9 mg/dL; p < 0.001) levels. The
underweight group was the oldest (mean age = 68.5 years; p < 0.001), had the lowest
proportion of females, and had the lowest mean SOFA score (8.2; p < 0.001) and number
of comorbidities (1.5; p = 0.014). This group had the lowest proportions of patients with
coronary artery disease (17.4%; p = 0.023), CKD (21.5%, p = 0.035), and diabetes mellitus
(32.1%; p < 0.001); highest proportions of patients with stroke (23.5%; p = 0.032), COPD
(16.1%; p = 0.005), cancer (26.4%; p = 0.002), and pneumonia (79.9%; p < 0.001); longest ICU
stay (14.0 days; p = 0.035); and lowest mean HbA1c (6.9%; p = 0.014) and capillary glucose
(172.1 mg/dL; p < 0.001) levels. The proportion of patients with coronary artery disease,
hypertension, and COPD and length of ICU stay were comparable in the validation cohort. 3.3. Survival Curve of Patients with Sepsis Classified According to the mNUTRIC Scores 3.3. Survival Curve of Patients with Sepsis Classified According to the mNUTRIC Scores Figure 1 shows that patients with sepsis who had low mNUTRIC scores had higher
90-day survival rates than those with high mNUTRIC scores in the construction cohort
(63.2% ± 2.1% vs. 54.1% ± 1.7%, respectively; p = 0.002) and validation (73.8% ± 1.9%
vs. 62.3% ± 2.2%; p < 0.001) cohort. Figure 2 shows the survival curves of patients with
sepsis in the three BMI subgroups. In the construction cohort, underweight patients with
lower mNUTRIC scores had a significantly higher 90-day survival rate (70.8% ± 4.2% vs. 54.2% ± 4.0%; p = 0.005), whereas overweight patients with lower mNUTRIC scores had a
higher 90-day survival rate (66.2% ± 3.6% vs. 55.2% ± 3.2%; p = 0.059). Normal-weight
patients did not show significantly different survival rates between the mNUTRIC score
groups (58.3% ± 3.3% vs. 54.9% ± 2.3%; p = 0.309). In the validation cohort, only over-
weight patients with lower mNUTRIC scores had significantly higher 90-day survival rates
(p < 0.001). 7 of 1
(58.3% ± 3.3% vs. 54.9% ± 2.3%; p = 0.309). In the validation cohort, only overweight pa
tients with lower mNUTRIC scores had significantly higher 90-day survival rates (p <
0.001). Figure 1. (A) Survival curve for the patients with sepsis in the medical intensive care unit according to the mNUTRIC
scores in the construction cohort; (B) survival curve for the patients with sepsis in the medical intensive care unit according
to the mNUTRIC scores in the validation cohort. Figure 1. (A) Survival curve for the patients with sepsis in the medical intensive care unit according to the mNUTRIC scores
in the construction cohort; (B) survival curve for the patients with sepsis in the medical intensive care unit according to the
mNUTRIC scores in the validation cohort. Figure 1. (A) Survival curve for the patients with sepsis in the medical intensive care unit according to the mNUTRIC
scores in the construction cohort; (B) survival curve for the patients with sepsis in the medical intensive care unit according
to the mNUTRIC scores in the validation cohort. Figure 1. (A) Survival curve for the patients with sepsis in the medical intensive care unit according to the mNUTRIC scores
in the construction cohort; (B) survival curve for the patients with sepsis in the medical intensive care unit according to the
mNUTRIC scores in the validation cohort. 7 of 17
IC
ng Nutrients 2021, 13, 1873
Figure 1. 3.3. Survival Curve of Patients with Sepsis Classified According to the mNUTRIC Scores (A) Survival curves for underweight patients with sepsis in the medical intensive care unit according to the
mNUTRIC scores in the construction cohort; (B) survival curves for normal-weight patients with sepsis in the medical
intensive care unit according to the mNUTRIC scores in the construction cohort; (C) survival curves for overweight pa-
tients with sepsis in the medical intensive care unit according to the mNUTRIC scores in the construction cohort; (D)
survival curves for underweight patients with sepsis in the medical intensive care unit according to the mNUTRIC scores
in the validation cohort; (E) survival curves for normal-weight patients with sepsis in the medical intensive care unit
Figure 2. (A) Survival curves for underweight patients with sepsis in the medical intensive care unit according to the
mNUTRIC scores in the construction cohort; (B) survival curves for normal-weight patients with sepsis in the medical
intensive care unit according to the mNUTRIC scores in the construction cohort; (C) survival curves for overweight patients
with sepsis in the medical intensive care unit according to the mNUTRIC scores in the construction cohort; (D) survival
curves for underweight patients with sepsis in the medical intensive care unit according to the mNUTRIC scores in the
validation cohort; (E) survival curves for normal-weight patients with sepsis in the medical intensive care unit according
to the mNUTRIC scores in the validation cohort; (F) survival curves for overweight patients with sepsis in the medical
intensive care unit according to the mNUTRIC scores in the validation cohort. Figure 2. (A) Survival curves for underweight patients with sepsis in the medical intensive care unit according to the
mNUTRIC scores in the construction cohort; (B) survival curves for normal-weight patients with sepsis in the medical
intensive care unit according to the mNUTRIC scores in the construction cohort; (C) survival curves for overweight pa-
tients with sepsis in the medical intensive care unit according to the mNUTRIC scores in the construction cohort; (D)
survival curves for underweight patients with sepsis in the medical intensive care unit according to the mNUTRIC scores
in the validation cohort; (E) survival curves for normal-weight patients with sepsis in the medical intensive care unit
Figure 2. 3.3. Survival Curve of Patients with Sepsis Classified According to the mNUTRIC Scores (A) Su
i th to the mNUTRIC scores in the validation cohort. Figure 2. (A) Survival curves for underweight patients with sepsis in the medical intensive care unit according to the
mNUTRIC scores in the construction cohort; (B) survival curves for normal-weight patients with sepsis in the medical
intensive care unit according to the mNUTRIC scores in the construction cohort; (C) survival curves for overweight pa-
tients with sepsis in the medical intensive care unit according to the mNUTRIC scores in the construction cohort; (D)
survival curves for underweight patients with sepsis in the medical intensive care unit according to the mNUTRIC scores
in the validation cohort; (E) survival curves for normal-weight patients with sepsis in the medical intensive care unit
Figure 2. (A) Survival curves for underweight patients with sepsis in the medical intensive care unit according to the
mNUTRIC scores in the construction cohort; (B) survival curves for normal-weight patients with sepsis in the medical
intensive care unit according to the mNUTRIC scores in the construction cohort; (C) survival curves for overweight patients
with sepsis in the medical intensive care unit according to the mNUTRIC scores in the construction cohort; (D) survival
curves for underweight patients with sepsis in the medical intensive care unit according to the mNUTRIC scores in the
validation cohort; (E) survival curves for normal-weight patients with sepsis in the medical intensive care unit according
to the mNUTRIC scores in the validation cohort; (F) survival curves for overweight patients with sepsis in the medical
intensive care unit according to the mNUTRIC scores in the validation cohort. urvival curves for underweight patients with sepsis in the medical intensive care unit according to the
urvival curves for underweight patients with sepsis in the medical intensive care unit according to the Figure 2. (A) Survival curves for underweight patients with sepsis in the medical intensive care unit a
Figure 2. (A) Survival curves for underweight patients with sepsis in the medical intensive care unit a Figure 2. 3.3. Survival Curve of Patients with Sepsis Classified According to the mNUTRIC Scores (A) Survival curves for underweight patients with sepsis in the medical intensive care unit according to the
mNUTRIC scores in the construction cohort; (B) survival curves for normal-weight patients with sepsis in the medical
intensive care unit according to the mNUTRIC scores in the construction cohort; (C) survival curves for overweight patients
with sepsis in the medical intensive care unit according to the mNUTRIC scores in the construction cohort; (D) survival
curves for underweight patients with sepsis in the medical intensive care unit according to the mNUTRIC scores in the
validation cohort; (E) survival curves for normal-weight patients with sepsis in the medical intensive care unit according
to the mNUTRIC scores in the validation cohort; (F) survival curves for overweight patients with sepsis in the medical
intensive care unit according to the mNUTRIC scores in the validation cohort. 3.4. Survival Curves for Patients with Sepsis According to the Different BMI Groups 3.4. Survival Curves for Patients with Sepsis According to the Different BMI Groups 3.4. Survival Curves for Patients with Sepsis According to the Different BMI Groups (A) Survival curves for the patients with sepsis in the medical intensive care unit according to the body mass
index (BMI) in the construction cohort; (B) survival curves for the patients with sepsis in the medical intensive care unit
according to the body mass index in the validation cohort. Figure 3. (A) Survival curves for the patients with sepsis in the medical intensive care unit according to the body mass
index (BMI) in the construction cohort; (B) survival curves for the patients with sepsis in the medical intensive care unit
according to the body mass index in the validation cohort. Figure 4. (A) Survival curves for the patients with sepsis in the medical intensive care unit who had low mNUTRIC scores
(≤5) according to body mass index in the construction cohort; (B) survival curves for the patients with sepsis in the medical
intensive care unit who had low mNUTRIC scores (≤5) according to body mass index in the validation cohort. * The 90-day
survival rate of the underweight group with low mNUTRIC scores was significantly higher than that of the normal-weight
group with low mNUTRIC scores (70.8% ± 4.2% vs. 58.3% ± 3.3%, respectively; p = 0.048). Figure 4. (A) Survival curves for the patients with sepsis in the medical intensive care unit who had low mNUTRIC scores
(≤5) according to body mass index in the construction cohort; (B) survival curves for the patients with sepsis in the medical
intensive care unit who had low mNUTRIC scores (≤5) according to body mass index in the validation cohort. * The 90-day
survival rate of the underweight group with low mNUTRIC scores was significantly higher than that of the normal-weight
group with low mNUTRIC scores (70.8% ± 4.2% vs. 58.3% ± 3.3%, respectively; p = 0.048). Table 3 shows that the construction cohort group with underweight patients who
had low mNUTRIC scores had the lowest proportion of females (underweight vs. normal-
weight vs. overweight: 29.7% vs. 34.6% vs. 47.9%, respectively; p = 0.041), lowest SOFA
scores on days 1, 3, and 7 (underweight vs. normal-weight vs. overweight: 6.1 ± 2.9
vs. 6.6 ± 3.1 vs. 7.6 ± 4.2, respectively, p = 0.039; 5.5 ± 2.2 vs. 6.5 ± 3.1 vs. 7.2 ± 3.5,
respectively, p = 0.006; and 4.5 ± 2.3 vs. 6.4 ± 3.5 vs. 6.2 ± 3.4, respectively, p = 0.001),
lowest proportion of patients with diabetes mellitus (underweight vs. 3.4. Survival Curves for Patients with Sepsis According to the Different BMI Groups Figure 3 shows that there was no significant difference in the survival among the three
BMI groups in the construction cohort (p = 0.372) and validation cohort (p = 0.132). Figure 4a
shows that the 90-day survival rate of the underweight group with low mNUTRIC scores
was significantly higher than that of the normal-weight group with low mNUTRIC scores
(70.8% ± 4.2% vs. 58.3% ± 3.3%, respectively; p = 0.048) in the construction cohort. The
survival rate was also higher in the overweight group than in the normal-weight group,
although the difference was not significant (66.2% ± 3.6% vs. 58.3% ± 3.3%; p = 0.199). In
the validation cohort, the 90-day survival rates were not significantly different among the
different BMI groups with low mNUTRIC scores (Figure 4b). 8 of 17
iffer- Nutrients 2021, 13, 1873 Figure 3. (A) Survival curves for the patients with sepsis in the medical intensive care unit according to the body mass
index (BMI) in the construction cohort; (B) survival curves for the patients with sepsis in the medical intensive care unit
according to the body mass index in the validation cohort. Figure 3. (A) Survival curves for the patients with sepsis in the medical intensive care unit according to the body mass
index (BMI) in the construction cohort; (B) survival curves for the patients with sepsis in the medical intensive care unit
according to the body mass index in the validation cohort. Figure 3. (A) Survival curves for the patients with sepsis in the medical intensive care unit according to the body mass
index (BMI) in the construction cohort; (B) survival curves for the patients with sepsis in the medical intensive care unit
according to the body mass index in the validation cohort. is in the medical intensive care unit according to the body mass Fi
3 (A) S
i
l
f
th
ti
t
ith i e a e u it a
o d Figure 3. (A) Survival curves for the patients with sepsis in the medical intensive care unit according to the body mass
index (BMI) in the construction cohort; (B) survival curves for the patients with sepsis in the medical intensive care unit
according to the body mass index in the validation cohort. Figure 3. 3.4. Survival Curves for Patients with Sepsis According to the Different BMI Groups normal-weight vs. overweight: 20.3% vs. 26.8% vs. 40.4%, respectively; p = 0.015), and highest proportion of
patients with pneumonia (underweight vs. normal-weight vs. overweight: 78.1% vs. 74.0%
vs. 54.4%, respectively; p = 0.007). The normal-weight group had the highest proportions of
patients with stroke (underweight vs. normal-weight vs. overweight: 12.5% vs. 13.4% vs. Nutrients 2021, 13, 1873 9 of 17 9 of 17 3.2%, respectively; p = 0.007) and cancer (underweight vs. normal-weight vs. overweight:
25% vs. 27.8% vs. 11.7%, p = 0.013), and the lowest mean HbA1c level (underweight vs. normal-weight vs. overweight: 7.0 ± 2.0 vs. 6.7 ± 1.7 vs. 7.7 ± 2.4, respectively; p = 0.027). In contrast, in the validation cohort group, there were higher proportions of patients with
comorbidities, such as coronary artery disease and hypertension, in the overweight group
with low mNUTRIC scores (Supplementary Table S2). 3.2%, respectively; p = 0.007) and cancer (underweight vs. normal-weight vs. overweight:
25% vs. 27.8% vs. 11.7%, p = 0.013), and the lowest mean HbA1c level (underweight vs. Table 3. Baseline characteristics and outcomes of patients with sepsis who had low modified NUTRIC scores. 3.4. Survival Curves for Patients with Sepsis According to the Different BMI Groups mNUTRIC Score ≤5
(n = 286)
Underweight
(n = 64)
Normal-Weight
(n = 128)
Overweight
(n = 94)
p
Demographic characteristics, mean ± SD
Age (years)
60.0 ± 16.1
61.8 ± 15.4
60.4 ± 15.9
58.2 ± 16.7
0.283
BMI, kg/m2
22.7 ± 5.2
16.4 ± 1.6
21.5 ± 1.9
28.6 ± 3.5
<0.001 ***
Sex (female), n (%)
108 (37.9)
19 (29.7)
44 (34.6)
45 (47.9)
0.041 *
Score, mean ± SD
APACHE II
18.1 ± 6.4
18.8 ± 6.4
18.1 ± 6.5
17.4 ± 6.3
0.276
Charlson comorbidity index
2.3 ± 2.1
2.6 ± 2.3
2.5 ± 2.3
2.0 ± 1.9
0.354
SOFA on day 1
6.8 ± 3.5
6.1 ± 2.9
6.6 ± 3.1
7.6 ± 4.2
0.039 *
SOFA on day 3
6.5 ± 3.1
5.5 ± 2.2
6.5 ± 3.1
7.2 ± 3.5
0.006 **
SOFA on day 7
5.9 ± 3.3
4.5 ± 2.3
6.4 ± 3.5
6.2 ± 3.4
0.001 **
mNUTRIC
4.1 ± 1.1
4.1 ± 1.1
4.1 ± 1.0
4.0 ± 1.1
0.550
Comorbidities (number)
1.1 ± 1.0
1.0 ± 0.9
1.2 ± 1.1
1.0 ± 1.1
0.112
Comorbidities, n (%)
Coronary artery disease
40 (14.0)
7 (10.9)
19 (15)
14 (14.9)
0.720
History of stroke
28 (9.8)
8 (12.5)
17 (13.4)
3 (3.2)
0.030 *
Hypertension
126 (44.4)
23 (35.9)
57 (45.2)
46 (48.9)
0.262
COPD
34 (11.9)
7 (10.9)
23 (18.1)
4 (4.3)
0.007 **
Cancer
62 (21.8)
16 (25.0)
35 (27.8)
11 (11.7)
0.013 *
CKD
42 (14.7)
5 (7.8)
21 (16.5)
16 (17.0)
0.206
Liver cirrhosis
19 (6.7)
4 (6.3)
7 (5.5)
8 (8.5)
0.669
Diabetes mellitus
85 (29.8)
13 (20.3)
34 (26.8)
38 (40.4)
0.015 *
Site of suspected infection, n (%)
Lung
198 (69.5)
50 (78.1)
94 (74.0)
54 (54.4)
0.007 **
UTI
56 (19.6)
12 (18.8)
24 (18.9)
20 (21.3)
0.889
Intraabdominal infection
17 (6.0)
1 (1.6)
12 (9.4)
4 (4.3)
0.066 †
Soft tissue infection
22 (7.7)
5 (7.8)
7 (5.5)
10 (10.6)
0369
Bacteraemia
17 (6.0)
3 (4.7)
7 (5.5)
7 (7.4)
0.741
Baseline glucose and HbA1c, mean ± SD
HbA1c (%) **
7.2 ± 2.1
7.0 ± 2.0
6.7 ± 1.7
7.7 ± 2.4
0.027 *
Glucose (mg/dL)
189.5 ± 136.7
165.8 ± 72.4
197.8 ± 184.2
194.5 ± 86.6
0.069 †
Admission days, mean ± SD
ICU days
12.9 ± 10.1
14.3 ± 11.2
12.3 ± 8.2
12.9 ± 11.7
0.399
LOS
29.5 ± 23.0
32.7 ± 27.3
26.5 ± 18.3
31.5 ± 25.2
0.630
Mortality, n (%)
7-day mortality
18 (6.3)
3 (4.7)
7 (5.5)
8 (8.5)
0.551
14-day mortality
34 (11.9)
8 (12.5)
13 (10.5)
13 (13.5)
0.708
28-day mortality
62 (21.8)
12 (18.8)
30 (23.6)
20 (21.3)
0.736
90-day mortality
98 (34.4)
17 (26.6)
50 (39.4)
31 (33.0)
0.200 a
*** p < 0.0001; ** p < 0.01; * p < 0.05; † p < 0.09; a underweight vs. 3.4. Survival Curves for Patients with Sepsis According to the Different BMI Groups normal-weight, p = 0.048. Abbreviations: SD, standard deviation;
mNUTRIC, modified nutrition risk in critically ill; BMI, body mass index; APACHE, acute physiology and chronic health evaluation;
SOFA, sequential organ failure assessment; COPD, chronic obstructive pulmonary disease; CKD, chronic kidney disease; UTI, urinary tract
infection; HbA1c, hemoglobin A1c; ICU, intensive care unit; LOS, length of stay. racteristics and outcomes of patients with sepsis who had low modified NUTRIC scores. p < 0.0001;
p < 0.01;
p < 0.05;
p < 0.09;
underweight vs. normal-weight, p = 0.048. Abbreviations: SD, standard deviation;
mNUTRIC, modified nutrition risk in critically ill; BMI, body mass index; APACHE, acute physiology and chronic health evaluation;
SOFA, sequential organ failure assessment; COPD, chronic obstructive pulmonary disease; CKD, chronic kidney disease; UTI, urinary tract
infection; HbA1c, hemoglobin A1c; ICU, intensive care unit; LOS, length of stay. Table 4 shows the odds ratios of the logistic regression analysis of risk factors for
90-day mortality in patients with sepsis who had low mNUTRIC scores. In Model 1,
the risk of mortality was reduced by nearly half (odds ratio (OR): 0.557; p = 0.082) in
underweight patients. In Model 2, after further adjusting for the SOFA scores, similar
trends were not observed for underweight patients. In Model 3, after further adjusting for Nutrients 2021, 13, 1873 10 of 17 10 of 17 comorbidities, the risk of mortality in underweight patients was still found to be reduced
by nearly half (OR: 0.529; p = 0.075). In addition, Model 3 showed that the incidence
of cancer significantly increased the risk of mortality by nearly four times (OR: 3.921;
p < 0.001). In Model 4, after considering pneumonia, underweight patients still showed
a risk of mortality that was decreased by nearly half (OR: 0.560; p = 0.085). In Model 5,
after HbA1c levels were considered, the effect of being underweight was found to be
insignificant. In contrast, HbA1c levels tended to decrease the risk of mortality by 21%
(OR: 0.792; p = 0.078), although this difference was not significant. In Model 6, after
considering all the above factors, only cancer was significantly associated with an increased
risk of mortality (OR: 3.833; p = 0.026). Table 4. Results (odds ratios) of the logistic regression analysis of risk factors for 90-day mortality in
patients with sepsis who had low modified NUTRIC scores. Table 4. 3.4. Survival Curves for Patients with Sepsis According to the Different BMI Groups Results (odds ratios) of the logistic regression analysis of risk factors for 90-day mortality in
patients with sepsis who had low modified NUTRIC scores. Model 1
Model 2
Model 3
Model 4
Model 5
Model 6
Intercept
0.598
0.330
0.437
0.651
1.578
1.433
Underweight
0.557 †
0.571
0.529 †
0.560 †
0.577
0.594
Overweight
0.756
0.712
0.926
0.735
0.853
0.905
Age
1.002
1.005
1.002
1.003
1.011
1.012
Sex (female), n (%)
0.950
0.924
0.936
0.951
0.661
0.544
Score
SOFA on day 1
1.062
1.002
Comorbidities
Coronary artery disease
0.785
0.593
COPD
0.948
1.927
Cancer
3.921 ***
3.833 *
DM
0.821
0.758
Site of suspected infection
Lung
0.841
0.717
Baseline glucose and HbA1c
HbA1c (%)
0.792 †
0.819
Goodness-of-fit
p
0.498
0.315
0.001 **
0.586
0.206
0.242
r2
0.012
0.021
0.086
0.013
0.054
0.10
*** p < 0.001; ** p < 0.01; * p < 0.05; † p < 0.09; abbreviations: mNUTRIC, modified nutrition risk in critically ill;
SOFA, sequential organ failure assessment; COPD, chronic obstructive pulmonary disease; DM, diabetes mellitus;
HbA1c, hemoglobin A1c. *** p < 0.001; ** p < 0.01; * p < 0.05; † p < 0.09; abbreviations: mNUTRIC, modified nutrition risk in critically ill;
SOFA, sequential organ failure assessment; COPD, chronic obstructive pulmonary disease; DM, diabetes mellitus;
HbA1c, hemoglobin A1c. In patients with high mNUTRIC scores, there was no significant difference in 90-
day mortality rates among the three BMI groups. Table 5 shows that the underweight
group had the lowest proportion of females (underweight vs. normal-weight vs. over-
weight: 30.6% vs. 39.7% vs. 52.3%, respectively; p = 0.003), lowest mean SOFA score
on day 1 (underweight vs. normal-weight vs. overweight: 9.8 ± 3.0 vs. 10.0 ± 3.5 vs. 11.0 ± 3.6, respectively; p = 0.007), lowest proportion of diabetes mellitus cases (under-
weight vs. normal-weight vs. overweight: 41.2% vs. 51.3% vs. 62.3%, respectively;
p = 0.006), highest proportion of pneumonia cases (underweight vs. normal-weight vs. overweight: 81.2% vs. 63.5% vs. 47.0%, respectively; p < 0.001), and lowest mean cap-
illary glucose level (underweight vs. normal-weight vs. overweight: 176.5 ± 74.8 vs. 216.2 ± 110.6 vs. 228.9 ± 108.1 mg/dL, respectively; p < 0.001). 11 of 17 Nutrients 2021, 13, 1873 Table 5. Baseline characteristics and outcomes of patients with sepsis who had high modified NUTRIC scores. 3.4. Survival Curves for Patients with Sepsis According to the Different BMI Groups mNUTRIC Score ≥6
(n = 513)
Underweight
(n = 85)
Normal-Weight
(n = 277)
Overweight
(n = 151)
p
Demographic characteristics, mean ± SD
Age (years)
71.1 ± 12.5
73.6 ± 12.9
71.6 ± 12.2
68.8 ± 12.5
0.006 **
BMI, kg/m2
22.9 ± 4.7
16.5 ± 1.5
21.8 ± 1.8
28.5 ± 3.5
<0.001 ***
Sex (female), n (%)
215 (41.9)
26 (30.6)
110 (39.7)
79 (52.3)
0.003 **
Score, mean ± SD
APACHE II
24.7 ± 8.4
27.4 ± 5.9
28.6 ± 6.8
28.8 ± 7.5
0.445
Charlson comorbidity index
2.6 ± 2.0
2.7 ± 2.0
2.8 ± 1.9
2.6 ± 1.8
0.633
SOFA on day 1
9.0 ± 3.8
9.8 ± 3.0
10.0 ± 3.5
11.0 ± 3.6
0.007 **
SOFA on day 3
8.4 ± 3.6
7.9 ± 3.0
8.3 ± 3.8
8.8 ± 3.6
0.118
SOFA on day 7
7.3 ± 3.8
6.9 ± 3.0
7.2 ± 3.9
7.9 ± 4.2
0.346
mNUTRIC
7.1 ± 1.0
7.1 ± 0.9
7.1 ± 1.0
7.1 ± 1.0
0.796
Comorbidities (number)
2.0 ± 1.1
1.9 ± 1.2
2.0 ± 1.1
2.0 ± 1.1
0.272
Comorbidities, n (%)
Coronary artery disease
164 (32.0)
19 (22.4)
98 (35.4)
47 (31.1)
0.327
History of stroke
119 (23.2)
27 (31.8)
62 (22.4)
30 (19.9)
0.103
Hypertension
318 (62.0)
50 (58.8)
168 (60.6)
100 (66.2)
0.423
COPD
82 (16.0)
17 (20.2)
48 (17.3)
17 (11.3)
0.142
Cancer
127 (25.0)
23 (27.4)
76 (27.8)
28 (17.7)
0.099
CKD
197 (38.4)
27 (31.8)
104 (37.5)
66 (43.7)
0.177
Liver cirrhosis
50 (9.7)
5 (5.9)
26 (9.4)
19 (12.6)
0.239
Diabetes mellitus
271 (52.8)
35 (41.2)
142 (51.3)
94 (62.3)
0.006 **
Site of suspected infection, n (%)
Lung
316 (61.6)
69 (81.2)
176 (63.5)
71 (47.0)
<0.001 ***
UTI
113 (22.0)
21 (24.7)
55 (19.9)
37 (24.5)
0.437
Intraabdominal infection
40 (7.8)
4 (4.7)
21 (7.6)
15 (9.9)
0.349
Soft tissue infection
18 (3.5)
3 (3.5)
7 (2.5)
8 (5.3)
0.330
Bacteremia
44 (8.6)
3 (3.5)
20 (7.2)
21 (13.9)
0.012 *
Baseline glucose and HbA1c, mean ± SD
HbA1c (%) **
7.3 ± 2.3
6.7 ± 2.1
7.3 ± 2.2
7.6 ± 2.5
0.078 †
Glucose (mg/dL)
213.8 ± 106.2
176.5 ± 74.8
216.2 ± 110.6
228.9 ± 108.1
0.001 **
Admission days, mean ± SD
ICU days
12.3 ± 9.5
13.8 ± 9.0
12.0 ± 9.6
11.9 ± 9.4
0.083 †
LOS
30.5 ± 31.0
37.7 ± 35.6
29.7 ± 31.6
28.0 ± 26.6
0.077 †
Mortality, n (%)
7-day mortality
83 (16.2)
10 (11.8)
46 (16.6)
27 (17.9)
0.454
14-day mortality
117 (22.8)
15 (17.6)
67 (24.2)
35 (23.2)
0.450
28-day mortality
164 (32.0)
23 (27.1)
89 (32.1)
52 (34.4)
0.504
90-day mortality
240 (46.8)
43 (50.6)
130 (46.9)
67 (44.4)
0.654
*** p < 0.001; ** p < 0.01; * p < 0.05, † p < 0.09; abbreviations: SD, standard deviation; mNUTRIC, modified nutrition risk in critically ill; BMI,
body mass index; APACHE, acute physiology and chronic health evaluation; SOFA, sequential organ failure assessment; COPD, chronic
obstructive pulmonary disease; CKD, chronic kidney disease; UTI, urinary tract infection; HbA1c, hemoglobin A1c; ICU, intensive care
unit; LOS, length of stay. 3.4. Survival Curves for Patients with Sepsis According to the Different BMI Groups seline characteristics and outcomes of patients with sepsis who had high modified NUTRIC scores. 3.5. Biomarkers of Patients with Sepsis ** p < 0.01; * p < 0.05; † p < 0.09; abbreviations: mNUTRIC, modified nutrition risk in critically ill; IL-6, interleukin 6; IL-1RA, interleukin 1
receptor antagonist; IFN-R, interferon receptor; IL-10, interleukin 10; IL-17, interleukin 17; TNF, tumor necrosis factor; HLA-DR, human
leukocyte antigen receptor. In the subgroup with patients who had high mNUTRIC scores, 112 patients underwent
biomarker analysis (underweight: n = 20; normal-weight: n = 61; overweight: n = 31). The
underweight group had the lowest mean HLA-DR level (underweight vs. normal-weight
vs. overweight: 77.3 ± 17.1 vs. 84.7 ± 17.3 vs. 88.0 ± 13.4, respectively; p = 0.031) (Table 7). In the subgroup with patients who had high mNUTRIC scores, 112 patients underwent
biomarker analysis (underweight: n = 20; normal-weight: n = 61; overweight: n = 31). The
underweight group had the lowest mean HLA-DR level (underweight vs. normal-weight
vs. overweight: 77.3 ± 17.1 vs. 84.7 ± 17.3 vs. 88.0 ± 13.4, respectively; p = 0.031) (Table 7). Table 7. Biomarkers of patients with sepsis who had high modified NUTRIC scores. Table 7. Biomarkers of patients with sepsis who had high modified NUTRIC scores. 3.5. Biomarkers of Patients with Sepsis 3.5. Biomarkers of Patients with Sepsis In the low mNUTRIC score subgroup, 53 patients underwent biomarker analysis
(underweight: n = 10; normal-weight: n = 18; and overweight: n = 25). The normal-weight
group had the lowest mean HLA-DR level on day 1 (underweight vs. normal-weight
vs. overweight: 94.3 ± 8.5 vs. 82.1 ± 19.0 vs. 94.3 ± 6.0, respectively; p = 0.007) and
day 3 (underweight vs. normal-weight vs. overweight: 91.8 ± 14.0 vs. 91.0 ± 5.0 vs. 93.2 ± 9.3, respectively; p = 0.047), and this was statistically significant. The normal-weight
group also had the lowest levels of IL-6 on day 1 (underweight vs. normal-weight vs. overweight: 174.3 ± 253.4 vs. 83.7 ± 164.4 vs. 104.5 ± 281.4, respectively; p = 0.076) and
day 3 (underweight vs. normal-weight vs. overweight: 188.1 ± 346.2 vs. 32.9 ± 47.7 vs. 129.7 ± 501.9, respectively; p = 0.072); IL-1RA on day 1 (underweight vs. normal-weight vs. overweight: 151.7 ± 123.4 vs. 49.5 ± 50.7 vs. 243.7 ± 394.9, respectively; p = 0.085) and Nutrients 2021, 13, 1873 12 of 17 12 of 17 day 3 (underweight vs. normal-weight vs. overweight: 147.3 ± 167.1 vs. 39.6 ± 49.8 vs. 77.6 ± 137.9, respectively; p = 0.061); and IFN-γ on day 1 (underweight vs. normal-weight
vs. overweight: 55.0 ± 77.3 vs. 9.3 ± 11.3 vs. 12.5 ± 15.5, respectively; p = 0.068) (Table 6). day 3 (underweight vs. normal-weight vs. overweight: 147.3 ± 167.1 vs. 39.6 ± 49.8 vs. 77.6 ± 137.9, respectively; p = 0.061); and IFN-γ on day 1 (underweight vs. normal-weight
vs. overweight: 55.0 ± 77.3 vs. 9.3 ± 11.3 vs. 12.5 ± 15.5, respectively; p = 0.068) (Table 6). Table 6. Biomarkers of patients with sepsis who had low modified NUTRIC scores. Table 6. Biomarkers of patients with sepsis who had low modified NUTRIC scores. 3.5. Biomarkers of Patients with Sepsis mNUTRIC Score ≤5
(n = 53)
Underweight
(n = 10)
Normal-Weight
(n = 18)
Overweight
(n = 25)
p
IL-6 on day 1 (10/18/25)
110.6 ± 239.9
174.3 ± 253.4
83.7 ± 164.4
104.5 ± 281.4
0.076 †
IL-6 on day 3 (10/17/24)
108.8 ± 376.3
188.1 ± 346.2
32.9 ± 47.7
129.7 ± 501.9
0.072 †
IL-6 on day 7 (10/15/21)
75.5 ± 241.4
51.5 ± 58.4
75.4 ± 155.9
87.0 ± 344.9
0.274
IL-1RA on day 1
160.4 ± 288.2
151.7 ± 123.4
49.5 ± 50.7
243.7 ± 394.9
0.085 †
IL-1RA on day 3
78.6 ± 126.6
147.3 ± 167.1
39.6 ± 49.8
77.6 ± 137.9
0.061 †
IL-1RA on day 7
84.9 ± 142.2
108.5 ± 133.1
104.4 ± 166.2
58.9 ± 130.3
0.351
IL-10 on day 1
30.3 ± 41.4
41.0 ± 53.2
21.7 ± 24.6
32.3 ± 46.0
0.826
IL-10 on day 3
35.7 ± 36.8
36.3 ± 51.2
17.3 ± 18.6
27.1 ± 39.7
0.784
IL-10 on day 7
32.0 ± 74.9
40.3 ± 81.9
42.9 ± 109.0
20.3 ± 32.0
0.792
IL-17 on day 1
10.6 ± 17.8
19.6 ± 29.3
6.9 ± 14.0
9.7 ± 13.6
0.124
IL-17 on day 3
12.5 ± 19.5
23.7 ± 30.4
7.8 ± 13.4
11.1 ± 16.3
0.099
IL-17 on day 7
9.5 ± 13.4
8.9 ± 8.1
9.8 ± 18.9
9.6 ± 11.0
0.901
TNF-α on day 1
33.5 ± 31.8
45.0 ± 29.8
20.7 ± 11.7
38.1 ± 39.4
0.159
TNF-α on day 3
29.8 ± 23.7
34.8 ± 26.8
30.4 ± 20.7
27.2 ± 25
0.276
TNF-α on day 7
23.1 ± 15.6
22.7 ± 18.6
22.9 ± 15.0
23.4 ± 15.3
0.963
IFN-R on day 1
19.4 ± 38.6
55.0 ± 77.3
9.3 ± 11.3
12.5 ± 15.5
0.068 †
IFN-R on day 3
23.1 ± 43.7
64.1 ± 83.9
12.2 ± 13.2
13.7 ± 19.2
0.600
IFN-R on day 7
21.0 ± 34.6
42.1 ± 60.8
16.1 ± 17.6
14.4 ± 23.0
0.677
HLA-DR on day 1
90.1 ± 13.5
94.3 ± 8.5
82.1 ± 19.0
94.3 ± 6.0
0.007 **
HLA-DR on day 3
92.2 ± 9.3
91.8 ± 14.0
91.0 ± 5.0
93.2 ± 9.3
0.047 *
HLA-DR on day 7
93.3 ± 7.7
95.0 ± 3.8
90.0 ± 8.5
94.0 ± 8.4
0.361
** p < 0.01; * p < 0.05; † p < 0.09; abbreviations: mNUTRIC, modified nutrition risk in critically ill; IL-6, interleukin 6; IL-1RA, interleukin 1
receptor antagonist; IFN-R, interferon receptor; IL-10, interleukin 10; IL-17, interleukin 17; TNF, tumor necrosis factor; HLA-DR, human
leukocyte antigen receptor. 4. Discussion In this study, the 90-day survival curves showed that underweight patients with low
mNUTRIC scores had significantly better survival rates than those with normal weights
and low mNUTRIC scores (70.8% vs. 58.3%, respectively; p = 0.048). In the regression
analysis, being underweight and the HbA1c levels predicted the survival of patients with
sepsis who had low mNUTRIC scores. An increase of 1% of HbA1c had a trend to decrease
20% mortality in patients with sepsis and low mNUTRIC scores (ORR:0.792, p = 0.078). Both these characteristics were protective factors for survival. Cancer was an independent
risk factor for survival, and patients with cancer had a nearly four-times higher risk of
mortality (βcancer = 3.921; p < 0.001) than those without cancer. Additionally, the mNUTRIC
scores predicted 90-day survival rates in critically ill patients, but not in those with normal
weights. This study revealed that when the mNUTRIC score of patients with sepsis was
≤5, being underweight was a protective factor and an indicator of survivability. We
also highlighted the limitations of the mNUTRIC scores of patients with sepsis who had
normal weights. The present study revealed that being underweight was associated with better survival
in patients with sepsis who had low mNUTRIC scores. Previous studies showed that being
underweight may be a risk factor for patients with sepsis [18–22,24], while one study
did not find a significant association between being underweight and outcomes in these
patients [41]. Nevertheless, these studies did not consider the heterogeneity of patients with
sepsis in the same BMI group. Most of them lacked baseline characteristics of clinical scores
(NUTRIC score, SOFA score, and APACHE II) and laboratory data (HbA1C and blood
glucose). In our analysis, the overall survival of patients with sepsis was not significantly
different from that of those with normal weights. However, there were differences in
the results of the subgroup analysis. Our results showed that better nutritional status of
patients with sepsis was correlated with improved survivability in underweight patients. We also found that patients with low mNUTRIC scores had the lowest SOFA scores on
days 1, 3, and 7 among the three BMI groups. The SOFA scores were used to evaluate
the severity of sepsis and predict the outcomes of patients with sepsis [1]. In model 1
of Table 4, the underweight group had a trend to have better survival in patients with
sepsis (0.557, p = 0.082). 3.5. Biomarkers of Patients with Sepsis mNUTRIC Score ≥6
(n = 112)
Underweight
(n = 20)
Normal-Weight
(n = 61)
Overweight (n = 31)
p
IL-6 on day 1 (20/61/31)
623.2 ± 2281.8
926.2 ± 3544.1
354.8 ± 1061.3
955.8 ± 2955.5
0.621
IL-6 on day 3 (20/54/26)
109.9 ± 320.5
61.8 ± 65.1
106.1 ± 200.1
154.8 ± 545.0
0.831
IL6 on day 7 (18/46/23)
50.1 ± 106.1
71.0 ± 195.9
40.5 ± 42.3
52.7 ± 100.8
0.875
IL-1RA on day 1
854.7 ± 2526.7
548.1 ± 1285.0
983.5 ± 3087.2
799.1 ± 1847.9
0.854
IL-1RA on day 3
361.7 ± 1474.9
748.2 ± 2951.2
279.3 ± 832.1
235.4 ± 613.7
0.936
IL-1RA on day 7
302.0 ± 1648.7
912.9 ± 3608.9
108.2 ± 165.1
211.6 ± 366.4
0.504
IL-10 on day 1
177.5 ± 446.8
130.2 ± 256.6
170.1 ± 448.4
222.5 ± 539.5
0.298
IL-10 on day 3
72.2 ± 222.2
47.5 ± 57.1
98.8 ± 297.1
36.0 ± 49.2
0.549
IL-10 on day 7
47.8 ± 168.7
45.9 ± 73.1
55.2 ± 225.8
34.5 ± 51.0
0.705
IL-17 on day 1
16.5 ± 34.2
29.4 ± 56.2
12.0 ± 18.2
17.1 ± 38.3
0.166
IL-17 on day 3
18.9 ± 40.0
40.1 ± 71.1
11.4 ± 22.0
18.3 ± 31.6
0.247
IL-17 on day 7
24.8 ± 98.7
66.2 ± 211.4
14.2 ± 25.3
13.7 ± 21.3
0.599
TNF-α on day 1
83.0 ± 143.3
53.8 ± 35.3
79.3 ± 108.1
109.3 ± 224.7
0.775
TNF-α on day 3
58.2 ± 87.0
35.6 ± 24.3
58.5 ± 87.5
74.6 ± 112.2
0.351
TNF-α on day 7
44.9 ± 38.3
37.5 ± 25.8
41.8 ± 30.6
56.8 ± 55.6
0.946
IFN-R on day 1
36.8 ± 88.9
64.1 ± 147.6
29.0 ± 72.4
34.4 ± 65.8
0.540
IFN-R on day 3
40.7 ± 142.9
122.9 ± 299.7
21.0 ± 47.3
18.4 ± 33.2
0.256
IFN-R on day 7
70.1 ± 389.5
233.3 ± 839.4
36.1 ± 100.0
10.1 ± 17.0
0.062 †
HLA-DR on day 1
84.3 ± 16.5
77.3 ± 17.1
84.7 ± 17.3
88.0 ± 13.4
0.031 *
HLA-DR on day 3
86.5 ± 15.4
78.1 ± 20.2
87.4 ± 14.7
90.7 ± 10.0
0.060 †
HLA-DR on day 7
92.8 ± 11.3
89.7 ± 14.5
92.0 ± 12.3
96.7 ± 3.3
0.218
* p < 0.05; † p < 0.09; abbreviations: mNUTRIC, modified nutrition risk in critically ill; IL-6, interleukin 6; IL-1RA, interleukin-1 receptor
antagonist; IFN-R, interferon receptor; IL-10, interleukin 10; IL-17, interleukin 17; TNF, tumor necrosis factor; HLA-DR, human leukocyte
antigen receptor. 3.5. Biomarkers of Patients with Sepsis Nutrients 2021, 13, 1873 13 of 17 13 of 17 4. Discussion This finding was consistent with that of previous studies, which showed that diabetes may
be associated with better outcomes [46]. In the validation cohort analysis, the 90-day survival curves did not show any sig-
nificant difference in survival rates among the different BMI groups with low mNUTRIC
scores. These findings were inconsistent with those of the construction cohort analysis. This is because the comorbidities differed between the construction and validation co-
horts. The normal-weight group with low mNUTRIC scores in the construction cohort
had higher cancer rates and stroke-associated mortality risk than the other BMI groups. In
contrast, the overweight group with low mNUTRIC scores in the validation cohort had a
higher cardiovascular-associated mortality risk than the other BMI groups. According to
a study by Flegal and colleagues, obesity may increase the risk of cardiovascular-related
mortality [20]. Therefore, the causes of mortality in the construction and validation cohorts
might have been different. There were only 46 patients in the underweight group with low
mNUTRIC scores. The sample size was too small to be statistically significant. p
y
g
In patients with high mNUTRIC scores, mortality rates were not significantly different
among the different BMI groups. Patients with high mNUTRIC scores had significantly
higher mortality rates than those with low mNUTRIC scores. Nearly half of the patients
with high mNUTRIC scores died within 90 days. Our analysis indicated that BMI did not
affect survival when patients with sepsis had poor nutritional status during the critical
phase. Moreover, the mNUTRIC scores predicted mortality in underweight and overweight
patients. However, in the normal-weight subgroup analysis, the mortality rates were
not significantly different between the high- and low-score groups, suggesting that the
mNUTRIC score was not a good predictor of survival in normal-weight patients. In the
validation cohort analysis, the normal-weight and overweight subgroup analyses were
consistent with this finding. g
Biomarker analysis further showed that patients in the normal-weight group with
low mNUTRIC scores had the lowest HLA-DR levels (82.1%) among the three BMI groups. According to our previous study, HLA-DR levels of <87.2% on day 1 indicated immune
dysfunction and predicted 28-day mortality [5]. In contrast, the mean HLA-DR level in
patients with high mNUTRIC scores was 84.3%, which was lower than that on day 1. Thus,
it was shown that patients with a high mortality risk had lower HLA-DR levels. 4. Discussion However, in adjusted model 2, the trend disappeared after we
adjusted it by SOFA scores. Thus, the survival of the underweight patients might have
been indicated by the low SOFA scores. Previous studies showed underweight patients
had lower SOFA scores compared to normal weight and obesity patients [42,43]. However,
the underweight group with lower SOFA scores did not have better survival in past studies. It may be related to heterogenecity of the underweight group. Unlike previous studies,
we categorized patients with sepsis by mNUTRIC score which was used to differentiate
nutrition status and severity. Our results revealed that underweight might not be a harmful
factor when patients with sepsis have better nutrition status. In addition, our analysis showed that cancer was an independent risk factor for sepsis-
related mortality. Our previous study also showed that patients with active cancer had
higher plasma IL-10 levels and mortality rates than those without cancer [3]. Normal-
weight patients with low mNUTRIC scores had the highest cancer incidence among the
three BMI groups (normal-weight vs. underweight vs. overweight: 25.0% vs. 27.8%,
vs. 11.7%, respectively; p = 0.013), and there was no significant difference between the
underweight and normal-weight patients (25.0% vs. 27.8%, respectively; p = 0.683). A
previous study showed that overweight and obesity may contribute to the risk of cancer [44]
and that cancer-related mortality may be associated with BMI changes after diagnosis [45]. Since our database lacked baseline BMI data before cancer, we could not determine whether
normal-weight and underweight patients with cancer were overweight before losing weight
due to cancer. However, a study by Flegal and colleagues revealed that being underweight
significantly increased non-cancer-related mortality but was not associated with cancer-
related mortality [20], suggesting that normal-weight patients had higher cancer mortality
rates than underweight patients. This could also explain why normal-weight patients had Nutrients 2021, 13, 1873 14 of 17 14 of 17 higher mortality rates than underweight patients despite similar cancer rates in our study. In addition, higher HbA1c levels were associated with improved odds of patient survival. This finding was consistent with that of previous studies, which showed that diabetes may
be associated with better outcomes [46]. higher mortality rates than underweight patients despite similar cancer rates in our study. In addition, higher HbA1c levels were associated with improved odds of patient survival. 4. Discussion Our
study also revealed that IL-6, IL-1RA, and IFN-γ levels were the lowest in the normal-
weight patients with low mNUTRIC scores. Previous studies have reported that the
levels of cytokines such as IL-1, IL-6, IFN-γ, and TNF-α increase during sepsis [9,10,39,47]
as an immune response to infection [10]. IL-1RA is a negative regulator of cytokine
storms [39]. Normal-weight patients with low NUTRIC scores had low cytokine levels on
day 1, suggesting a poor immune response against sepsis, which may lead to poor survival. One strength of the present study was that it revealed that being underweight was an
indicator of survivability in patients whose mNUTRIC scores were ≤5. Being underweight
may not always be harmful if patients have optimal clinical nutritional status. Another
strength was that we revealed the limitations of the mNUTRIC score for normal-weight
patients. The development of novel or improved nutritional evaluation tools in the future
is warranted to benefit normal-weight patients. In addition, we included both the con-
struction cohort and validation cohort to confirm our findings and analyze the differences. Finally, we used biomarkers to validate our findings. This study validated our previous
study on the HLA-DR level cut-off for predicting survival. To conclude, we used nutri-
tional status to detect patients with high survival and analyzed biomarkers to explain the
pathophysiological mechanisms. Nevertheless, this study had some limitations. First, the study used body weights
measured during ICU admission. BMI might be misclassified because of a lack of fluid
balance adjustment [23]. Second, BMI does not represent complete body components,
especially body fat. Previous studies have shown that obesity may reduce the risk of Nutrients 2021, 13, 1873 15 of 17 mortality via the secretion of leptin and soluble TNF receptor 2 during sepsis in adipose
tissues [26]. Third, finger sugar, HbA1c, and biological data were not available for the
validation cohort. Further studies are needed to clarify the relationship between complete
body components and sepsis. In addition, the survival of patients with sepsis may be
affected by multiple factors, such as age, sex, comorbidity, infection sites, and antibiotics. Although we used controls for age, sex, multiple comorbidities, and infection sites, we
could not make a direct causal correlation between nutritional status and survival, which
would require further evidence. Since this was a single-center retrospective study, we need
further prospective trials to confirm our results. 5. Conclusions The major contribution of our study was that it revealed that being underweight may
not always be harmful if patients have optimal clinical nutritional status. Our findings
have important implications for clinicians, as they can guide the application of therapeutic
strategies and provide a better understanding of the underlying mechanisms of sepsis-
related mortality. Supplementary Materials: The following are available online at https://www.mdpi.com/article/10
.3390/nu13061873/s1, Table S1: Baseline characteristics and outcomes of patients with sepsis in the
validation cohort of the study, and Table S2: Baseline characteristics and outcomes of patients with
sepsis who had low modified NUTRIC scores in the validation cohort of the study. Author Contributions: Conceptualization and methodology: Y.-H.T. and W.-F.F.; acquisition, anal-
ysis, or interpretation of data: Y.-H.T., C.-Y.L., Y.-M.C., Y.-P.C., K.-Y.H., Y.-C.C. (Ya-Chun Chang),
and H.-C.C., K.-T.H., Y.-C.C. (Yung-Che Chen), Y.-H.W., C.-C.W., M.-C.L., and W.-F.F.; drafting the
manuscript and revising it critically for important intellectual content: Y.-H.T. and W.-F.F. All authors
have read and agreed to the published version of the manuscript. Funding: This research was supported in part by grants from the Chang Gung Memorial Hospital
(CMRPG8K1671, Y.H. Tsai, CMRPG8B1063, CMRPG8H1171, and CMRPG8J0422 to W.F. Fang and
CFRPG8H0431 to K.Y. Hung). Institutional Review Board Statement: The study was conducted according to the guidelines of the
Declaration of Helsinki and approved by the Institutional Review Board of Chang Gung Memorial
Hospital (ID: 202001696B0C501; approval date: 21 December 2020). Informed Consent Statement: Informed consent was obtained from 165 participants involved in the
study from whom blood samples were collected. Informed Consent Statement: Informed consent was obtained from 165 participants involved in the
study from whom blood samples were collected. Data Availability Statement: The data presented in this study are available on request from the
corresponding author. The data are not publicly available due to patients’ privacy. Acknowledgments: We thank the ICU staff and clinicians who participated in this study for their
support. We also thank the Biostatistics Center of Kaohsiung Chang Gung Memorial Hospital for the
statistical consultations. Conflicts of Interest: The authors declare no conflict of interest. Conflicts of Interest: The authors declare no conflict of interest. 3.
Fang, W.-F.; Chen, Y.-M.; Lin, C.-Y.; Huang, K.-T.; Kao, H.-C.; Fang, Y.-T.; Huang, C.-H.; Chang, Y.-T.; Wang, Y.-H.; Wang, C.-C.
Immune profiles and clinical outcomes between sepsis patients with or without active cancer requiring admission to intensive
care units. PLoS ONE 2017, 12, e0179749. 4.
Yang, Y.; Yang, K.S.; Hsann, Y.M.; Lim, V.; Ong, B.C. The effect of comorbidity and age on hospital mortality and length of stay in
patients with sepsis. J. Crit. Care 2010, 25, 398–405. [CrossRef] References 1. Singer, M.; Deutschman, C.S.; Seymour, C.W.; Shankar-Hari, M.; Annane, D.; Bauer, M.; Bellomo, R.; Bernard, G.R.; Chiche, J.-D.;
Coopersmith, C.M. The third international consensus definitions for sepsis and septic shock (Sepsis-3). JAMA 2016, 315, 801–810. [CrossRef] [PubMed] [
] [
]
2. Phua, J.; Koh, Y.; Du, B.; Tang, Y.-Q.; Divatia, J.V.; Tan, C.C.; Gomersall, C.D.; Faruq, M.O.; Shrestha, B.R.; Binh, N.G. Management
of severe sepsis in patients admitted to Asian intensive care units: Prospective cohort study. BMJ 2011, 342, d3245. [CrossRef]
[PubMed] 3. Fang, W.-F.; Chen, Y.-M.; Lin, C.-Y.; Huang, K.-T.; Kao, H.-C.; Fang, Y.-T.; Huang, C.-H.; Chang, Y.-T.; Wang, Y.-H.; Wang, C.-C. Immune profiles and clinical outcomes between sepsis patients with or without active cancer requiring admission to intensive
care units. PLoS ONE 2017, 12, e0179749. 16 of 17 16 of 17 Nutrients 2021, 13, 1873 5. Hung, K.-Y.; Chen, Y.-M.; Wang, C.-C.; Wang, Y.-H.; Lin, C.-Y.; Chang, Y.-T.; Huang, K.-T.; Lin, M.-C.; Fang, W.-F. Insufficient
nutrition and mortality risk in septic patients admitted to ICU with a focus on immune dysfunction. Nutrients 2019, 11, 367. [CrossRef] [PubMed] [
] [
]
6. Brascher, J.M.M.; Peres, W.A.F.; Padilha, P.C. Use of the modified "Nutrition Risk in the critically ill"
the death of critically ill patients. Clin. Nutr. ESPEN 2020, 35, 162–166. [CrossRef] [PubMed] y
p
[
] [
]
7. Fang, W.-F.; Chen, Y.-M.; Wang, Y.-H.; Huang, C.-H.; Hung, K.-Y.; Fang, Y.-T.; Chang, Y.-C.; Lin, C.-Y.; Chang, Y.-T.; Chen, H.-C. Incorporation of dynamic segmented neutrophil-to-monocyte ratio with leukocyte count for sepsis risk stratification. Sci. Rep. 2019, 9, 1–9. 8. Fang, W.-F.; Fang, Y.-T.; Huang, C.-H.; Chen, Y.-M.; Chang, Y.-C.; Lin, C.-Y.; Hung, K.-Y.; Chang, Y.-T.; Chen, H.-C.; Huang, K.-T. Risk factors and associated outcomes of ventilator-associated events developed in 28 days among sepsis patients admitted to
intensive care unit. Sci. Rep. 2020, 10, 1–10. [CrossRef] p
9. Fang, W.-F.; Douglas, I.S.; Chen, Y.-M.; Lin, C.-Y.; Kao, H.-C.; Fang, Y.-T.; Huang, C.-H.; Chang, Y.-T.; Huang, K.-T.; Wang,
Y.-H. Development and validation of immune dysfunction score to predict 28-day mortality of sepsis patients. PLoS ONE 2017,
12, e0187088. [CrossRef] 10. Grimble, R.F. Basics in clinical nutrition: Main cytokines and their effect during injury and sepsis. Clin. Nutr. ESPEN 2008, 6,
e289–e292. [CrossRef] 11. Scicluna, B.P.; Van Vught, L.A.; Zwinderman, A.H.; Wiewel, M.A.; Davenport, E.E.; Burnham, K.L.; Nürnberg, P.; Schultz, M.J.;
Horn, J.; Cremer, O.L. References Flegal, K.M.; Graubard, B.I.; Williamson, D.F.; Gail, M.H. Cause-specific excess deaths associated with u
and obesity JAMA 2007 298 2028 2037 [CrossRef] [PubMed] p
M.; Graubard, B.I.; Williamson, D.F.; Gail, M.H. Cause-specific excess deaths associated with underweight, o
ity. JAMA 2007, 298, 2028–2037. [CrossRef] [PubMed] 20. Flegal, K.M.; Graubard, B.I.; Williamson, D.F.; Gail, M.H. Cause-specific excess deaths associated with u
and obesity. JAMA 2007, 298, 2028–2037. [CrossRef] [PubMed] 21. Gu, D.; He, J.; Duan, X.; Reynolds, K.; Wu, X.; Chen, J.; Huang, G.; Chen, C.-S.; Whelton, P.K. Body weight and mortality among
men and women in China. JAMA 2006, 295, 776–783. [CrossRef] [PubMed] 22. Flegal, K.M.; Graubard, B.I.; Williamson, D.F.; Gail, M.H. Excess deaths associated with underweight, overweight, and obesity. JAMA 2005, 293, 1861–1867. [CrossRef] 23. Zhou, Q.; Wang, M.; Li, S.; Zhang, J.; Ma, Q.; Ding, Y.; Ge, H.; Shen, N.; Zheng, Y.; Sun, Y. Impact of body mass index on survival of
medical patients with sepsis: A prospective cohort study in a university hospital in China. BMJ Open 2018, 8, e021979. [CrossRef] 23. Zhou, Q.; Wang, M.; Li, S.; Zhang, J.; Ma, Q.; Ding, Y.; Ge, H.; Shen, N.; Zheng, Y.; Sun, Y. Impact of body mass index on survival of
medical patients with sepsis: A prospective cohort study in a university hospital in China. BMJ Open 2018, 8, e021979. [CrossRef]
24. Weng, L.; Fan, J.; Yu, C.; Guo, Y.; Bian, Z.; Wei, Y.; Yang, L.; Chen, Y.; Du, H.; Chang, L.; et al. Body-mass index and long-term risk
of sepsis-related mortality: A population-based cohort study of 0.5 million Chinese adults. Crit. Care 2020, 24, 534. [CrossRef]
[PubMed] 24. Weng, L.; Fan, J.; Yu, C.; Guo, Y.; Bian, Z.; Wei, Y.; Yang, L.; Chen, Y.; Du, H.; Chang, L.; et al. Body-mass index and long-term risk
of sepsis-related mortality: A population-based cohort study of 0.5 million Chinese adults. Crit. Care 2020, 24, 534. [CrossRef]
[PubMed] 25. Barr, J.; Hecht, M.; Flavin, K.E.; Khorana, A.; Gould, M.K. Outcomes in critically ill patients before and after the implementation
of an evidence-based nutritional management protocol. Chest 2004, 125, 1446–1457. [CrossRef] [PubMed] 26. Pepper, D.J.; Demirkale, C.Y.; Sun, J.; Rhee, C.; Fram, D.; Eichacker, P.; Klompas, M.; Suffredini, A.F.; Kadri, S.S. Does Obesity
Protect Against Death in Sepsis? A Retrospective Cohort Study of 55,038 Adult Patients. Crit. Care Med. 2019, 47, 643–650. [CrossRef] 27. References Classification of patients with sepsis according to blood genomic endotype: A prospective cohort study. Lancet Respir. Med. 2017, 5, 816–826. [CrossRef] p
12. McClave, S.A.; Taylor, B.E.; Martindale, R.G.; Warren, M.M.; Johnson, D.R.; Braunschweig, C.; McCarthy, M.S.; Davanos, E.; Rice,
T.W.; Cresci, G.A. Guidelines for the provision and assessment of nutrition support therapy in the adult critically ill patient:
Society of Critical Care Medicine (SCCM) and American Society for Parenteral and Enteral Nutrition (A.S.P.E.N.). J. Parenter. Enter. Nutr. 2016, 40, 159–211. [CrossRef] [PubMed] 13. Mukhopadhyay, A.; Henry, J.; Ong, V.; Leong, C.S.-F.; Teh, A.L.; van Dam, R.M.; Kowitlawakul, Y. Association of modified
NUTRIC score with 28-day mortality in critically ill patients. Clin. Nutr. 2017, 36, 1143–1148. [CrossRef] 14. Correia, M.I.T.; Campos, A.C.L.; Study, E.C. Prevalence of hospital malnutrition in Latin America: The multicenter ELAN study. Nutrition 2003, 19, 823–825. [CrossRef] Nutrition 2003, 19, 823–825. [CrossRef]
15. Waitzberg, D.L.; Caiaffa, W.T.; Correia, M.I.T. Hospital malnutrition: The Brazilian national survey (IBRANUTRI): A study of
4000
ti
t
N t iti
2001 17 573 580 [C
R f] Nutrition 2003, 19, 823 825. [CrossRef]
15. Waitzberg, D.L.; Caiaffa, W.T.; Correia, M.I.T. Hospital malnutrition: The Brazilian national survey (IBRANUTRI): A study of
4000 patients. Nutrition 2001, 17, 573–580. [CrossRef] Caiaffa, W.T.; Correia, M.I.T. Hospital malnutrition: The Brazilian national survey (IBRANUTRI): A stud
utrition 2001, 17, 573–580. [CrossRef] 15. Waitzberg, D.L.; Caiaffa, W.T.; Correia, M.I.T. Hospital malnutrition: The Brazilian national survey (I
4000 patients. Nutrition 2001, 17, 573–580. [CrossRef] p
16. Kamath, S.K.; Lawler, M.; Smith, A.E.; Kalat, T.; Olson, R. Hospital malnutrition: A 33-hospital screening study. J. Am. Diet. Assoc. 1986, 86, 203. ,
,
17. O’Flynn, J.; Peake, H.; Hickson, M.; Foster, D.; Frost, G. The prevalence of malnutrition in hospitals can be reduced: Results from
three consecutive cross-sectional studies. Clin. Nutr. 2005, 24, 1078–1088. [CrossRef] [PubMed] 17. O’Flynn, J.; Peake, H.; Hickson, M.; Foster, D.; Frost, G. The prevalence of malnutrition in hospitals
three consecutive cross-sectional studies. Clin. Nutr. 2005, 24, 1078–1088. [CrossRef] [PubMed] 18. Xue, Z.; Yu, J.; Higashikuchi, T.; Compher, C. Does Low Body Mass Index Predict Mortality in Asian Hospitalized Patients? J. Parenter. Enter. Nutr. 2020, 44, 722–728. [CrossRef] 19. Lee, S.; Kang, J.; Jo, Y.; Kim, K.; Lee, J.; Lee, J.; Rim, K. Underweight is associated with mortality in patients with severe sepsis and
septic shock. Intensive Care Med. Exp. 2015, 3, 1. [CrossRef] p
p
20. References Mukhopadhyay, A.; Kowitlawakul, Y.; Henry, J.; Ong, V.; Leong, C.S.-F.; Tai, B.C. Higher BMI is associated with reduced mortality
but longer hospital stays following ICU discharge in critically ill Asian patients. Clin. Nutr. ESPEN 2018, 28, 165–170. [CrossRef]
[PubMed] 28. Wang, S.; Liu, X.; Chen, Q.; Liu, C.; Huang, C.; Fang, X. The role of increased body mass index in outcomes of sepsis: A systematic
review and meta-analysis. BMC Anesthesiol. 2017, 17, 118. [CrossRef] 17 of 17 17 of 17 Nutrients 2021, 13, 1873 29. Pepper, D.J.; Sun, J.; Welsh, J.; Cui, X.; Suffredini, A.F.; Eichacker, P.Q. Increased body mass index and adjusted mortality in ICU
patients with sepsis or septic shock: A systematic review and meta-analysis. Crit. Care 2016, 20, 181. [CrossRef] p
p
p
y
y
30. Nguyen, A.T.; Tsai, C.L.; Hwang, L.Y.; Lai, D.; Markham, C.; Patel, B. Obesity and Mortality, Length of Stay and Hospital Cost p
p
p
y
y
30. Nguyen, A.T.; Tsai, C.L.; Hwang, L.Y.; Lai, D.; Markham, C.; Patel, B. Obesity and Mortality, Length of Stay and Hospital Cost
among Patients with Sepsis: A Nationwide Inpatient Retrospective Cohort Study. PLoS ONE 2016, 11, e0154599. [CrossRef] 31. Sakr, Y.; Alhussami, I.; Nanchal, R.; Wunderink, R.G.; Pellis, T.; Wittebole, X.; Martin-Loeches, I.; Francois, B.; Leone, M.; Vincent,
J.L.; et al. Being Overweight Is Associated With Greater Survival in ICU Patients: Results From the Intensive Care Over Nations
Audit. Crit. Care Med. 2015, 43, 2623–2632. [CrossRef] 32. Prescott, H.C.; Chang, V.W.; O’Brien, J.M., Jr.; Langa, K.M.; Iwashyna, T.J. Obesity and 1-year outcomes in older Americans with
severe sepsis. Crit. Care Med. 2014, 42, 1766–1774. [CrossRef] p
33. Ng, P.Y.; Eikermann, M. The obesity conundrum in sepsis. BMC Anesthesiol. 2017, 17, 147. [CrossRef] 34. Papadimitriou-Olivgeris, M.; Aretha, D.; Zotou, A.; Koutsileou, K.; Zbouki, A.; Lefkaditi, A.; Sklavou, C.; Marangos, M.; Fligou, F. The Role of Obesity in Sepsis Outcome among Critically Ill Patients: A Retrospective Cohort Analysis. Biomed Res. Int. 2016,
2016, 5941279. [CrossRef] [PubMed] 35. Seidell, J.C.; Verschuren, W.M.; Van Leer, E.M.; Kromhout, D. Overweight, underweight, and mortality: A prospective study of
48287 men and women. Arch. Intern. Med. 1996, 156, 958–963. [CrossRef] [PubMed] .I.; Tamim, H.M.; Rishu, A.H.; Bouchama, A.; Khedr, M.K.; Feinstein, D.; Parrillo, J.E.; Wood, K.E.; Keenan 36. Arabi, Y.M.; Dara, S.I.; Tamim, H.M.; Rishu, A.H.; Bouchama, A.; Khedr, M.K.; Feinstein, D.; Parrillo, J.E.; Wood, K.E.; Keenan,
S.P.; et al. References Clinical characteristics, sepsis interventions and outcomes in the obese patients with septic shock: An international
multicenter cohort study. Crit. Care 2013, 17, R72. [CrossRef] [PubMed] S.P.; et al. Clinical characteristics, sepsis interventions and outcomes in the obese patients with septic shock: An international
multicenter cohort study. Crit. Care 2013, 17, R72. [CrossRef] [PubMed] y
37. Gaulton, T.G.; MacNabb, C.M.; Mikkelsen, M.E.; Agarwal, A.K.; Sante, S.C.; Shah, C.V.; Gaieski, D.F. A retrospective cohort
study examining the association between body mass index and mortality in severe sepsis. Intern. Emerg. Med. 2015, 10, 471–479. [CrossRef] [PubMed] [
] [
]
38. Kuperman, E.F.; Showalter, J.W.; Lehman, E.B.; Leib, A.E.; Kraschnewski, J.L. The impact of obesity on sepsis mortality: A
retrospective review. BMC Infect. Dis. 2013, 13, 377. [CrossRef] 39. Martino, J.L.; Stapleton, R.D.; Wang, M.; Day, A.G.; Cahill, N.E.; Dixon, A.E.; Suratt, B.T.; Heyland, D.K. Extreme obesity and
outcomes in critically ill patients. Chest 2011, 140, 1198–1206. [CrossRef] y
p
40. Fajgenbaum, D.C.; June, C.H. Cytokine storm. N. Engl. J. Med. 2020, 383, 2255–2273. [CrossRef] 41. Wurzinger, B.; Dunser, M.W.; Wohlmuth, C.; Deutinger, M.C.; Ulmer, H.; Torgersen, C.; Schmittinger, C.A.; Grander, W.; Hasibeder,
W.R. The association between body-mass index and patient outcome in septic shock: A retrospective cohort study. Wien. Klin. Wochenschr. 2010, 122, 31–36. [CrossRef] [PubMed] [
] [
]
42. Sakr, Y.; Madl, C.; Filipescu, D.; Moreno, R.; Groeneveld, J.; Artigas, A.; Reinhart, K.; Vincent, J.-L. Obesity is associated with
increased morbidity but not mortality in critically ill patients. Intensive Care Med. 2008, 34, 1999–2009. [CrossRef] [PubMed] 42. Sakr, Y.; Madl, C.; Filipescu, D.; Moreno, R.; Groeneveld, J.; Artigas, A.; Reinhart, K.; Vincent, J.-L. Obesity is associated with
increased morbidity but not mortality in critically ill patients. Intensive Care Med. 2008, 34, 1999–2009. [CrossRef] [PubMed]
43. Hutagalung, R.; Marques, J.; Kobylka, K.; Zeidan, M.; Kabisch, B.; Brunkhorst, F.; Reinhart, K.; Sakr, Y. The obesity paradox in
surgical intensive care unit patients. Intensive Care Med. 2011, 37, 1793–1799. [CrossRef] [PubMed] increased morbidity but not mortality in critically ill patients. Intensive Care Med. 2008, 34, 1999–2009. [CrossRef] [PubMed]
43. Hutagalung, R.; Marques, J.; Kobylka, K.; Zeidan, M.; Kabisch, B.; Brunkhorst, F.; Reinhart, K.; Sakr, Y. The obesity paradox in
surgical intensive care unit patients. Intensive Care Med. 2011, 37, 1793–1799. [CrossRef] [PubMed] 43. References Hutagalung, R.; Marques, J.; Kobylka, K.; Zeidan, M.; Kabisch, B.; Brunkhorst, F.; Reinhart, K.; Sakr, Y
surgical intensive care unit patients. Intensive Care Med. 2011, 37, 1793–1799. [CrossRef] [PubMed] 44. Brown, K.F.; Rumgay, H.; Dunlop, C.; Ryan, M.; Quartly, F.; Cox, A.; Deas, A.; Elliss-Brookes, L.; Gavin, A.; Hounsome, L. The
fraction of cancer attributable to modifiable risk factors in England, Wales, Scotland, Northern Ireland, and the United Kingdom
in 2015. Br. J. Cancer 2018, 118, 1130–1141. [CrossRef] [PubMed] 45. Shachar, S.S.; Williams, G.R. The Obesity Paradox in Cancer—Moving Beyond BMI. Cancer Epidemiol Biomarkers Prev. 2017, 26,
13–16. [CrossRef] 46. Esper, A.M.; Moss, M.; Martin, G.S. The effect of diabetes mellitus on organ dysfunction with sepsis: An epidemiological study. Crit. Care 2009, 13, R18. [CrossRef] 47. Kolyva, A.S.; Zolota, V.; Mpatsoulis, D.; Skroubis, G.; Solomou, E.E.; Habeos, I.G.; Assimakopoulos, S.F.; Goutzourelas, N.;
Kouretas, D.; Gogos, C.A. The role of obesity in the immune response during sepsis. Nutr. Diabetes 2014, 4, e137. [CrossRef]
|
https://openalex.org/W4229008030
|
https://www.research-collection.ethz.ch/bitstream/20.500.11850/545714/2/tc-16-1697-2022.pdf
|
English
| null |
Modelling supraglacial debris-cover evolution from the single-glacier to the regional scale: an application to High Mountain Asia
|
The cryosphere
| 2,022
|
cc-by
| 19,042
|
ETH Library ETH Library Author(s): ( )
Compagno, Loris
; Huss, Matthias
; Miles, Evan Stewart; McCarthy, Michael James; Zekollari, Harry
; Dehecq, Amaury
;
Pellicciotti, Francesca; Farinotti, Daniel Correspondence: Loris Compagno (compagno@vaw.baug.ethz.ch) Correspondence: Loris Compagno (compagno@vaw.baug.ethz.ch) Received: 4 August 2021 – Discussion started: 27 August 2021 Received: 4 August 2021 – Discussion started: 27 August 2021
Revised: 29 March 2022 – Accepted: 1 April 2022 – Published: 6 May 2022 Received: 4 August 2021 – Discussion started: 27 August 2021
Revised: 29 March 2022 – Accepted: 1 April 2022 – Published: 6 May 2022 Revised: 29 March 2022 – Accepted: 1 April 2022 – Published: 6 May 2022 Abstract. Currently, about 12 %–13 % of High Mountain
Asia’s glacier area is debris-covered, which alters its sur-
face mass balance. However, in regional-scale modelling
approaches, debris-covered glaciers are typically treated as
clean-ice glaciers, leading to a bias when modelling their fu-
ture evolution. Here, we present a new approach for mod-
elling debris area and thickness evolution, applicable from
single glaciers to the global scale. We derive a parameteri-
zation and implement it as a module into the Global Glacier
Evolution Model (GloGEMflow), a combined mass-balance
ice-flow model. The module is initialized with both glacier-
specific observations of the debris’ spatial distribution and
estimates of debris thickness. These data sets account for the
fact that debris can either enhance or reduce surface melt de-
pending on thickness. Our model approach also enables rep-
resenting the spatiotemporal evolution of debris extent and
thickness. We calibrate and evaluate the module on a se-
lected subset of glaciers and apply GloGEMflow using dif-
ferent climate scenarios to project the future evolution of all
glaciers in High Mountain Asia until 2100. Explicitly ac-
counting for debris cover has only a minor effect on the pro-
jected mass loss, which is in line with previous projections. Despite this small effect, we argue that the improved pro-
cess representation is of added value when aiming at captur-
ing intra-glacier scales, i.e. spatial mass-balance distribution. Depending on the climate scenario, the mean debris-cover fraction is expected to increase, while mean debris thickness
is projected to show only minor changes, although large lo-
cal thickening is expected. To isolate the influence of explic-
itly accounting for supraglacial debris cover, we re-compute
glacier evolution without the debris-cover module. We show
that glacier geometry, area, volume, and flow velocity evolve
differently, especially at the level of individual glaciers. This
highlights the importance of accounting for debris cover and
its spatiotemporal evolution when projecting future glacier
changes. 1
Introduction In High Mountain Asia (HMA), debris-covered and clean-
ice glaciers are losing mass due to climate change (Brun
et al., 2017; Zemp et al., 2019; Shean et al., 2020; Hugonnet
et al., 2021). Since the atmosphere is expected to warm fur-
ther (Lee et al., 2021), more glacier mass is expected to be
lost (Marzeion et al., 2020; Rounce et al., 2020). Understand-
ing how sensitive HMA glaciers are to changes in climate is
crucial to quantify the future glacier evolution in the area. A key unknown is the present and future influence of
supraglacial debris cover in moderating melt rates for the
12 %–13 % of HMA’s glacier area that is presently covered
by debris (Herreid and Pellicciotti, 2020). A better under- Funding acknowledgement: 184634 - Process-based modelling of global glacier changes (PROGGRES) (SNF)
869304 - PROjecTing sEa-level rise : from iCe sheets to local implicaTions (EC) This page was generated automatically upon download from the ETH Zurich Research Collection. For more information, please consult the Terms of use. The Cryosphere, 16, 1697–1718, 2022
https://doi.org/10.5194/tc-16-1697-2022
© Author(s) 2022. This work is distributed under
the Creative Commons Attribution 4.0 License. Modelling supraglacial debris-cover evolution from the
single-glacier to the regional scale: an application to
High Mountain Asia Loris Compagno1,2, Matthias Huss1,2,3, Evan Stewart Miles2, Michael James McCarthy2, Harry Zekollari4,5,1,2,
Amaury Dehecq1,2, Francesca Pellicciotti2, and Daniel Farinotti1,2
1Laboratory of Hydraulics, Hydrology and Glaciology (VAW), ETH Zurich, Zurich, Switzerland
2Swiss Federal Institute for Forest, Snow and Landscape Research (WSL), Birmensdorf, Switzerland
3Department of Geosciences, University of Fribourg, Fribourg, Switzerland
4Department of Geoscience and Remote Sensing, Delft University of Technology, Delft, the Netherlands
5Laboratoire de Glaciologie, Université libre de Bruxelles, Brussels, Belgium Loris Compagno1,2, Matthias Huss1,2,3, Evan Stewart Miles2, Michael James McCarthy2, Harry Zekollari4,5,1,2,
Amaury Dehecq1,2, Francesca Pellicciotti2, and Daniel Farinotti1,2
1Laboratory of Hydraulics, Hydrology and Glaciology (VAW), ETH Zurich, Zurich, Switzerland
2Swiss Federal Institute for Forest, Snow and Landscape Research (WSL), Birmensdorf, Switzerland
3Department of Geosciences, University of Fribourg, Fribourg, Switzerland
4Department of Geoscience and Remote Sensing, Delft University of Technology, Delft, the Netherlands
5Laboratoire de Glaciologie, Université libre de Bruxelles, Brussels, Belgium L. Compagno et al.: Regional-scale modelling of supraglacial debris However, for thicker, continuous debris layers, the increased
isolation layer allows for high debris surface temperatures
(often > 15 ◦C), thereby increasing both the outgoing long-
wave radiation and the turbulent energy fluxes directed away
from the surface (e.g. Nicholson and Benn, 2006; Steiner
et al., 2018). This results in a reduced and delayed conduc-
tion of energy to the glacier ice, leading to a progressive re-
duction in melt with increasing debris thickness (e.g. Østrem,
1959; Reznichenko et al., 2010a; Anderson and Anderson,
2016; Rounce et al., 2021). Regional and global models with various levels of com-
plexity have been used to simulate HMA’s future glacier
evolution (see Marzeion et al., 2020, for a model inter-
comparison). The models use different methodologies for
computing ablation, accumulation, or geometry changes but
rarely take into account the debris cover and its spatiotem-
poral evolution. An exception is the study by Kraaijenbrink
et al. (2017) that presented the first HMA glacier projec-
tions explicitly accounting for the effect of supraglacial de-
bris. However, the study neither considered an evolution of
debris extent and thickness in the future nor modelled ice
flow explicitly or used glacier-specific mass-balance data for
calibration. Glacier-specific studies considering debris-cover
evolution exist (e.g. Jouvet et al., 2011; Rowan et al., 2015;
Kienholz et al., 2017; Scherler and Egholm, 2020; Verhae-
gen et al., 2020), as well as theoretical and process-based
modelling studies (Anderson and Anderson, 2016; Ferguson
and Vieli, 2021), but the majority are based on higher-order
ice-flow models and require rather extensive observational
data. Thus, the corresponding methods are hardly applicable
at larger scales. Since
glaciers
are
presently
far
from
equilibrium
(Marzeion et al., 2018; Zekollari et al., 2020; Miles et al.,
2021), their debris cover is evolving through time (Stokes
et al., 2007; Bhambri et al., 2011; Bolch et al., 2011; Shukla
and Qadir, 2016; Tielidze et al., 2020). Indeed, medial
moraines and debris patches – which are formed by the ac-
cumulation and transport of debris – tend to grow and to ex-
pand laterally with increasing ablation (Anderson, 2000; Jou-
vet et al., 2011; Rowan et al., 2015; Kienholz et al., 2017;
Wirbel et al., 2018; Verhaegen et al., 2020). Additionally,
ice-marginal moraines, which may become unstable when
glaciers retreat, can supply the ice surface with additional de-
bris (Van Woerkom et al., 2019). L. Compagno et al.: Regional-scale modelling of supraglacial debris 1698 By combining estimates of sub-debris melt with surface
temperature inversion methods, Rounce et al. (2021) re-
cently presented the first global estimate of supraglacial de-
bris thickness distribution on glaciers. The estimate refers
to about 2008, but debris thickness evolves through time. The few direct observations available indicate a debris-cover
thickening in the last decades (e.g. Gibson et al., 2017; Ver-
haegen et al., 2020), most probably related to the negative
mass balances induced by ongoing climate change as well
as to the resulting glacier thinning and decelerated ice flow
(Verhaegen et al., 2020; Anderson et al., 2021a). Supraglacial
ice cliffs and ponds might additionally contribute to this, as
they enhance local ablation of debris-covered glaciers (Sakai
et al., 2000, 1998; Ragettli et al., 2016; Miles et al., 2018)
and evolve as well (Narama et al., 2017; Watson et al., 2017;
Chand and Watanabe, 2019; Buri et al., 2021; Ferguson and
Vieli, 2021). standing is necessary to accurately predict future water avail-
ability; to assess impacts on irrigation, hydropower, and both
public and private usage of water (Biemans et al., 2019;
Farinotti et al., 2019b; Fyffe et al., 2019; Immerzeel et al.,
2020; Miles et al., 2021); to anticipate hotspots of hazards
such as ice-dammed or proglacial lakes (Emmer et al., 2014;
Zheng et al., 2021); or to project the glaciers’ contribution to
sea-level rise (Edwards et al., 2021). standing is necessary to accurately predict future water avail-
ability; to assess impacts on irrigation, hydropower, and both
public and private usage of water (Biemans et al., 2019;
Farinotti et al., 2019b; Fyffe et al., 2019; Immerzeel et al.,
2020; Miles et al., 2021); to anticipate hotspots of hazards
such as ice-dammed or proglacial lakes (Emmer et al., 2014;
Zheng et al., 2021); or to project the glaciers’ contribution to
sea-level rise (Edwards et al., 2021). The presence of debris at the ice surface has the effect of
reducing the surface albedo and increasing the net short-wave
radiation (Owen et al., 2003; Reid and Brock, 2010). When
debris is particularly thin and/or patchy, this excess energy
can be readily conducted to the ice, thus enhancing melt rates
(Østrem, 1959; Reznichenko et al., 2010a; Fyffe et al., 2020). Published by Copernicus Publications on behalf of the European Geosciences Union. Published by Copernicus Publications on behalf of the European Geosciences Union. L. Compagno et al.: Regional-scale modelling of supraglacial debris L. Compagno et al.: Regional-scale modelling of supraglacial debris based nature of the procedure, the energy-balance model and
the Østrem curves (see below) are not explicitly calibrated
but use model parameter that are based on literature values. The debris thickness maps are evaluated using a large amount
of available in situ data (148 007 data points on 13 glaciers)
on debris thickness, showing good agreement (see McCarthy
et al., 2021). To model surface mass balance, the energy-
balance model was run at randomly chosen points on the sur-
face of each glacier and with randomly chosen debris thick-
nesses and debris properties within expected physical ranges. To generate the Østrem curves, in a first step, the energy-
balance model was run at randomly chosen points on the sur-
face of each considered glacier and with debris thicknesses
and debris properties randomly chosen within expected phys-
ical ranges. These Østrem curves are expressed as mass balance, glacier evolution, and ice-flow velocity, which
allows us to assess the importance of accounting for debris-
cover evolution in regional studies. 2
Data To model the evolution all 95 536 glaciers contained in the
Randolph Glacier Inventory version 6.0 (RGI 6.0; RGI Con-
sortium, 2017) for HMA over the 21st century, different data
sets are used (see Fig. 1). 2.1
Glacier geometry Rounce et al., 2021). For our purposes, the spatially
distributed debris-cover information is divided into elevation
bands of 10 m, whilst the Østrem curves were directly added
into our mass-balance module (see Sect. 3.1). where b is the local surface mass balance (m w.e. a−1), h is
the debris thickness (m), and idebris (m w.e. a−1) and kdebris
(m) are glacier-specific calibration parameters without spe-
cific physical meaning. The mean and 95 % confidence inter-
val for idebris are −1.86 and [−7.62, −0.09], respectively. For
kdebris the equivalent values are 0.10 and [0.01, 0.22]. Note
that Eq. (1) has similarities with the hyper-fit model of An-
derson and Anderson (2016) and Anderson et al. (2021a, b),
although we note that the two approaches differ in the num-
ber of parameters and their interpretation. In a second step,
the mass balances inferred by Miles et al. (2021) were used
together with the fitted Østrem curves (Eq. 1) for each el-
evation (i.e. assuming that englacial and basal mass bal-
ance is negligible) to determine the debris thickness maps
used in this study. The so-obtained information represents
the supraglacial debris conditions for the period 2000–2016. With the method described above, McCarthy et al. (2021) es-
timate a mean debris thickness for the debris-covered part of
all glaciers in HMA of 0.34 m (with an uncertainty between
0.15 and 0.76 m). The uncertainties are asymmetric because
surface mass balance is less sensitive to debris thickness as
debris thickness increases and are in line with other stud-
ies (e.g. Rounce et al., 2021). For our purposes, the spatially
distributed debris-cover information is divided into elevation
bands of 10 m, whilst the Østrem curves were directly added
into our mass-balance module (see Sect. 3.1). L. Compagno et al.: Regional-scale modelling of supraglacial debris As a consequence, glaciers
with negative mass balances tend to increase their debris-
cover fractions through time (Stokes et al., 2007; Bhambri
et al., 2011; Bolch et al., 2011; Shukla and Qadir, 2016;
Tielidze et al., 2020). In the Karakoram region, instead, pos-
itive and negative debris-cover changes have offset one an-
other during the past 40 years (Herreid et al., 2015). This is
most probably the consequence of the neutral or even slightly
positive mass balance in the region (Gardelle et al., 2013;
Farinotti et al., 2020). g
Here, we present a new debris area and thickness evolu-
tion module applicable to both individual glaciers and the
regional to global scale. The module is included into the
Global Glacier Evolution Model (GloGEMflow), a combined
mass-balance (Huss and Hock, 2015) ice-flow (Zekollari
et al., 2019) model. We calibrate and extensively evaluate
the debris-cover module and showcase its applicability for all
glaciers in High Mountain Asia. We focus on the future evo-
lution of debris cover and determine the impacts that explic-
itly modelling debris-cover evolution has on transient glacier
evolution. To do so, we model all HMA glaciers between
2000 and 2100. The modelling is based on five Shared So-
cioeconomic Pathways (SSP119, SSP126, SSP245, SSP370,
and SSP585) from the sixth phase of the Coupled Model
Intercomparison Project (CMIP6), and the results are com-
pared to model runs that do not explicitly account for debris
cover. We discuss the resulting differences in terms of glacier For glaciers with negative mass balances, the debris also
progressively expands up-glacier (Stokes et al., 2007), to-
gether with the rise in the equilibrium line altitude (ELA). Indeed, the mass-balance profile of a debris-covered glacier
may have a local minimum at mid-elevations, especially if
the ice is clean (i.e. not covered by debris) at that eleva-
tion. This fosters the expansion of the debris-cover fraction
through the melt-out of englacial debris transported by the
glacier’s ice flow (Stokes et al., 2007; Rowan et al., 2015). https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022 1699 2.1
Glacier geometry We use glacier outlines from RGI 6.0 (RGI Consortium,
2017), which is a global inventory of glacier outlines. For
HMA glaciers, the RGI outlines are based on remote sensing
data acquired between 1998 and 2013. For the ice thickness,
we use the consensus estimate by Farinotti et al. (2019a),
which is based on an ensemble of models using characteris-
tics of the glacier surface (e.g. slope and surface velocities)
and principles of ice-flow dynamics for ice thickness inver-
sion. For the modelling, the geometry is simplified by sub-
dividing each glacier into elevation bands of 10 m, includ-
ing tributary glaciers (Huss and Hock, 2015) (i.e. they are
not treated separately). In this elevation-dependent represen-
tation, the transversal glacier bed shape is parameterized as-
suming a glacier cross-section that has the form of an isosce-
les trapezoid with 45◦base angles (see Zekollari et al., 2019,
for more details). b = idebris · kdebris
h + kdebris
,
(1) b = idebris · kdebris
h + kdebris
, b = idebris · kdebris
h + kdebris
, (1) where b is the local surface mass balance (m w.e. a−1), h is
the debris thickness (m), and idebris (m w.e. a−1) and kdebris
(m) are glacier-specific calibration parameters without spe-
cific physical meaning. The mean and 95 % confidence inter-
val for idebris are −1.86 and [−7.62, −0.09], respectively. For
kdebris the equivalent values are 0.10 and [0.01, 0.22]. Note
that Eq. (1) has similarities with the hyper-fit model of An-
derson and Anderson (2016) and Anderson et al. (2021a, b),
although we note that the two approaches differ in the num-
ber of parameters and their interpretation. In a second step,
the mass balances inferred by Miles et al. (2021) were used
together with the fitted Østrem curves (Eq. 1) for each el-
evation (i.e. assuming that englacial and basal mass bal-
ance is negligible) to determine the debris thickness maps
used in this study. The so-obtained information represents
the supraglacial debris conditions for the period 2000–2016. With the method described above, McCarthy et al. (2021) es-
timate a mean debris thickness for the debris-covered part of
all glaciers in HMA of 0.34 m (with an uncertainty between
0.15 and 0.76 m). The uncertainties are asymmetric because
surface mass balance is less sensitive to debris thickness as
debris thickness increases and are in line with other stud-
ies (e.g. 2.2
Debris cover and Østrem curves For each glacier with an area > 2 km2 with debris cover
(i.e. 6115 glaciers in total; see RGI Consortium, 2017), the
debris coverage is represented by a debris-cover mask gener-
ated using Landsat scenes acquired between 2013 and 2017
(Scherler et al., 2018), as well as spatially distributed debris-
cover thickness maps and glacier-specific Østrem curves
(i.e. a function that characterizes the relation between debris-
cover thicknesses and melt rates; Østrem, 1959). The debris thickness maps are based on McCarthy et al. (2021), who used a simplified surface mass-balance inver-
sion procedure similar to Ragettli et al. (2015) and Rounce
et al. (2018). In a nutshell, the procedure uses the principle
of mass conservation to infer local glacier mass balance from
surface velocities and thinning rates and then iteratively ad-
justs the debris thickness to ensure consistency between the
so-inferred mass balance and the output of an energy-balance
model driven by meteorological data. More specifically, the
procedure uses digital elevation models (DEMs), glacier ice
thickness, surface velocity, debris proprieties, and meteoro-
logical forcing data as input and uses them to calculate ice
flux divergence and ice thinning rates. The debris thickness
is then adjusted until modelled and observed ice-melt rates
agree within a prescribed tolerance. Due to the physically To calibrate and evaluate the parameterizations used for
describing the evolution of both debris area and thickness
(see Sect. 3.2), we use multiple Hexagon and Landsat satel-
lite images acquired between 1973–1976 and 1987–2019, re-
spectively. The Hexagon images (Maurer and Rupper, 2015),
available as scan of raw film images from the US Geologi-
cal Survey, were georeferenced and orthorectified following
the methodology of Dehecq et al. (2020). Since the Hexagon
images are monochromatic, debris cover has been delineated
manually for 31 glaciers distributed through HMA. We use
glaciers with sufficient contrast between debris and clean ice, https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022 1700 L. Compagno et al.: Regional-scale modelling of supraglacial debris igure 1. (a) Extent of HMA glaciers (white) as per Randolph Glacier Inventory version 6 (RGI; RGI Consortium, 2017). The three
GI regions (Central Asia, South Asia West, and South Asia East) are shown by blueish, reddish, and greenish colours, respectively. econd-order regions are labelled individually. Three glaciers are highlighted to illustrate glacier-specific model results (red circles
umbers). 2.2
Debris cover and Østrem curves (b, c, d) Map of the three highlighted glaciers with their mean 2000–2016 debris thickness given by colours (scale in pane
lacier outlines and debris thickness are from RGI Consortium (2017) and McCarthy et al. (2021), respectively. For each glacier, V i
lacier ice volume according to Farinotti et al. (2019a); A is the glacier area according to RGI 6.0; Adebris is the debris-covered area
debris is the mean debris-cover thickness with superscript and subscript values indicating its estimated confidence interval (note tha
tter is not symmetric; cf. Sect. 2.2). (e, f, g) Glacier hypsometry (area per 10 m elevation band) and debris-covered area distributio
nventory date; n is the number of glaciers within each region (RGI Consortium, 2017). Map background source: Natural Earth. Figure 1. (a) Extent of HMA glaciers (white) as per Randolph Glacier Inventory version 6 (RGI; RGI Consortium, 2017). The three main
RGI regions (Central Asia, South Asia West, and South Asia East) are shown by blueish, reddish, and greenish colours, respectively. RGI
second-order regions are labelled individually. Three glaciers are highlighted to illustrate glacier-specific model results (red circles with
numbers). (b, c, d) Map of the three highlighted glaciers with their mean 2000–2016 debris thickness given by colours (scale in panel b). Glacier outlines and debris thickness are from RGI Consortium (2017) and McCarthy et al. (2021), respectively. For each glacier, V is the
glacier ice volume according to Farinotti et al. (2019a); A is the glacier area according to RGI 6.0; Adebris is the debris-covered area; and
hdebris is the mean debris-cover thickness with superscript and subscript values indicating its estimated confidence interval (note that the
latter is not symmetric; cf. Sect. 2.2). (e, f, g) Glacier hypsometry (area per 10 m elevation band) and debris-covered area distribution at
inventory date; n is the number of glaciers within each region (RGI Consortium, 2017). Map background source: Natural Earth. 2004–2009, 2010–2014, and 2015–2019. Each multi-date
composite is visually checked, then a debris–ice transition
threshold is chosen automatically with an Otsu routine (Otsu,
1979). By using images stemming from different epochs and
through suitable selection (see Sect. 3.2), the area covered by
debris is identified for 68 glaciers, again scattered throughout
HMA. All 31 glaciers for which debris cover is identified on little to absent shadows on the glacier surface, and with-
out snow on the ablation zone. L. Compagno et al.: Regional-scale modelling of supraglacial debris ules (i.e. the ones dealing with mass balance, ice flow, and
debris cover) are presented. the Hexagon images are also covered by the Landsat data. All
glaciers are divided into two sets. The first set (termed S1) is
composed of 55 glaciers where debris is identified on Land-
sat images and 18 glaciers where the debris is identified from
Hexagon imagery. The second set (termed S2) is composed
of 11 glaciers where debris is identified on both Landsat and
Hexagon images. This division into two sets is done to en-
sure independence between data used in the calibration and
the evaluation of the debris-evolution module. 2.4
Climate For forcing GloGEMflow between 1979 and 2020, we use
2 m temperature and precipitation data of the European
Centre for Medium-Range Weather Forecasts Reanalysis
(ERA5) (Hersbach et al., 2019). For the future (2020–2100),
we use 53 members of the CMIP6 ensemble (Eyring et al.,
2016) from 12 different general circulation models (GCMs),
covering 5 SSPs (5 members for SSP119 and 12 members for
all other SSPs). Both data sets have a monthly resolution. To
ensure consistency between past and future, a de-biasing pro-
cedure is applied that adjusts the GCMs to the ERA5 data set
(see Huss and Hock, 2015, for details). This procedure ap-
plies a set of additive and multiplicative corrections to adjust
the long-term mean difference and the short-term variability
in the coarse-resolution GCMs (with a horizontal resolution
of about 100 km) to the high-resolution ERA5 data (with a
horizontal resolution of about 30 km). (
adebris
z,t
= az,t · g
if g < 1.65,
adebris
z,t
= az,t · 1.65
if g > 1.65,
(2) (2) where adebris
z,t
(m w.e. a−1) is ablation of debris-covered ice
and az,t (m w.e. a−1) is ablation of bare ice at elevation z and
time t. The factor g (which acts as a factor enhancing ab-
lation due to debris) depends on debris-cover thickness hz,t
(m) and the glacier-specific parameter kdebris (see Eq. 1), used
by McCarthy et al. (2021) to fit the Østrem curve; g can be
expressed as follows: g =
( (kdebris+hcrit)
hz,t+kdebris ,
if hz,t > heff,
(kdebris+hcrit)
heff+kdebris · hz,t
heff + heff−hz,t
heff
,
if hz,t < heff,
(3) (3) (3) where hcrit is the critical debris thickness (m), i.e. the debris
thickness for which ice melt beneath debris is identical to the
melt of bare ice (Reznichenko et al., 2010b) and heff is the de-
bris thickness for which the enhancement of melt is maximal. Here, we use hcrit = 0.036 m and heff = 0.016 m, which are
the means for hcrit and heff as determined from observations 2.3
Mass balance To calibrate the mass-balance module of GloGEMflow, we
rely on glacier-wide geodetic volume changes available for
2000–2019 (Hugonnet et al., 2021). These volume changes
were obtained from surface-elevation changes determined by
using stereo images from the Advanced Spaceborne Thermal
Emission and Reflection Radiometer (ASTER). The volume
changes are converted into mass changes by using a con-
stant density conversion factor of 850 kg m−3 (Huss, 2013). The data set provides a volume change estimate for all in-
dividual glaciers. It covers ∼99.8 % of the regional glacier
area (Hugonnet et al., 2021). For the remaining ∼0.2 %, data
from a nearby glacier are chosen by following the same pro-
cedure as described in Compagno et al. (2021). To evaluate
the mass-balance module, we use independent data from in
situ observations provided by the World Glacier Monitoring
Service for 21 glaciers (WGMS, 2020). A degree-day model (Hock, 2003) is used to compute ab-
lation. Ice, firn, and snow are differentiated by using a dif-
ferent degree-day factor (DDF), with a ratio between DDFice
and DDFsnow of 2.0 and a ratio between DDFice and DDFfirn
of 1.5. The air temperature lapse rate, used to determine
temperature for each elevation band of the glacier surface,
is computed from temperature fields at distinct geopotential
heights provided by the ERA5 data set (see Compagno et al.,
2021, for more details). For debris-covered ice, melt enhancement and reduction
due to thin and thick debris cover, respectively, are accounted
for. This is done by applying a glacier-specific Østrem curve
(see Sect. 2.2) that relates ablation (a) under debris to debris
thickness (h) using Eq. (1), while the standard GloGEMflow-
calculated ablation (without debris) is used for h = 0. In Glo-
GEMflow, the relation between ablation and debris cover is
applied to each elevation z and each time step t and can be
expressed as 3.1
Mass balance Accumulation is computed by summing solid precipitation,
which is determined by applying a local temperature thresh-
old of 1.5 ◦C (with a linear transition between liquid and
solid precipitation in the 0.5 and 2.5 ◦C range) to the ERA5
grid cell closest to the glacier. To account for the precipitation
increase with elevation, we apply a lapse rate of 0.015 % m−1
(for consistency, it is the same as Huss and Hock, 2015). For
glaciers with an elevation range over 1000 m, precipitation is
reduced in the uppermost quarter with an exponential func-
tion to account for reduced moisture content in the air and
stronger wind erosion (see Huss and Hock, 2015, for details). 2.2
Debris cover and Østrem curves For the Landsat satellite im-
ages, debris is identified automatically following the multi-
date composite approach of Scherler et al. (2018). We apply
this method to the combined multi-sensor Landsat archive
in Google Earth Engine for additional epochs with sufficient
Landsat acquisitions: 1987–1991, 1994–1999, 1999–2003, https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022 1701 3.2
Debris-cover evolution where γz,t is the fraction of debris cover in elevation band z
at time t, bz,t is the mass balance at elevation z at time t,
and B(t−9,t) is the 10-year moving average of the glacier-
wide mass balance, evaluated between years t −9 and t;
clateral is a regional debris-cover extension parameter which
is calibrated to minimize the difference between observed
and computed lateral expansion of debris (see Sect. 4.2). The
first term of Eq. (4) accounts for pre-existing debris cover,
while the second term describes the rate of debris expansion. The latter is proportional to the local mass balance abs(bz,t),
which is generally negative where debris cover is present,
thus accounting for the expected increase in lateral debris for
locations with higher melt rates (Jouvet et al., 2011; Stokes
et al., 2007; Bhambri et al., 2011; Bolch et al., 2011; Shukla
and Qadir, 2016; Tielidze et al., 2020). where γz,t is the fraction of debris cover in elevation band z
at time t, bz,t is the mass balance at elevation z at time t,
and B(t−9,t) is the 10-year moving average of the glacier-
wide mass balance, evaluated between years t −9 and t;
clateral is a regional debris-cover extension parameter which
is calibrated to minimize the difference between observed
and computed lateral expansion of debris (see Sect. 4.2). The
first term of Eq. (4) accounts for pre-existing debris cover,
while the second term describes the rate of debris expansion. The latter is proportional to the local mass balance abs(bz,t),
which is generally negative where debris cover is present,
thus accounting for the expected increase in lateral debris for
locations with higher melt rates (Jouvet et al., 2011; Stokes
et al., 2007; Bhambri et al., 2011; Bolch et al., 2011; Shukla
and Qadir, 2016; Tielidze et al., 2020). The evolution of debris-cover extent and thickness is parame-
terized by accounting for three main processes: (1) the lateral
expansion of debris cover within individual elevation bands,
which is meant to mimic the observed lateral expansion of
medial moraines and debris patches; (2) the debris up-glacier
expansion, which describes the progressive appearance of de-
bris at higher elevation when the ELA rises; and (3) debris
thickness evolution, which accounts for the progressive accu-
mulation of debris on the surface due to insufficient export by
ice flow (see Fig. 4). 3.2.1
Lateral expansion of debris cover by 11 local studies across HMA (Khan, 1989; Mattson and
Gardner, 1989; Kayastha et al., 2000; Tangborn and Rana,
2000; Mihalcea et al., 2006; Hagg et al., 2008; Wei et al.,
2010; Dobhal et al., 2013; Juen et al., 2014; Sharma et al.,
2016; Groos et al., 2018; see Table S1 in the Supplement). by 11 local studies across HMA (Khan, 1989; Mattson and
Gardner, 1989; Kayastha et al., 2000; Tangborn and Rana,
2000; Mihalcea et al., 2006; Hagg et al., 2008; Wei et al.,
2010; Dobhal et al., 2013; Juen et al., 2014; Sharma et al.,
2016; Groos et al., 2018; see Table S1 in the Supplement). In Eq. (2), g is constrained to a maximal value of 1.65,
which corresponds to the highest observed melt enhancement
factor reported in the 11 local studies (see Table S1). For
glacier-specific Østrem curves and examples of hcrit and heff,
see Figs. 3 and S1 in the Supplement. The lateral expansion of an already existing debris layer
(e.g. medial and ice-marginal moraines, as well as isolated
debris patches) is linked to the local mass balance (Stokes
et al., 2007; Bhambri et al., 2011; Bolch et al., 2011; Shukla
and Qadir, 2016; Tielidze et al., 2020). We describe the pro-
cess of lateral expansion on a yearly time step by In Eq. (2), g is constrained to a maximal value of 1.65,
which corresponds to the highest observed melt enhancement
factor reported in the 11 local studies (see Table S1). For
glacier-specific Østrem curves and examples of hcrit and heff,
see Figs. 3 and S1 in the Supplement. γz,t = γz,t−1 + abs(bz,t) · B(t−9,t) · (−1) · γz,t−1
· clateral,if z < ELA,
(4) (4) 3
Methods GloGEMflow is a combined mass-balance and ice-flow
model, extended with a new component for debris-cover evo-
lution for this study. The general workflow of this study is
illustrated in Fig. 2. In the following sections, the three mod- https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022 L. Compagno et al.: Regional-scale modelling of supraglacial debris L. Compagno et al.: Regional-scale modelling of supraglacial debris 1702 Figure 2. Study overview. The grey numbers correspond to the sections of this paper. The “METHODS” column also depicts the different
modules included in GloGEMflow. The violet arrows show the iterations between the modules during the calibration. Figure 2. Study overview. The grey numbers correspond to the sections of this paper. The “METHODS” column also depicts the different
modules included in GloGEMflow. The violet arrows show the iterations between the modules during the calibration. 3.2.2
Up-glacier expansion of debris cover For glaciers with negative mass balances, debris cover has
been observed to progressively expand up-glacier (Deline
and Orombelli, 2005; Stokes et al., 2007). We assume that
this is related to the rise in the ELA and in the melt-out of
debris in areas that transit from the accumulation to the abla-
tion zone (Anderson, 2000). As Eq. (4) does not permit sim-
ulating the expansion of debris to new elevation bands, we
parameterize this process as Figure 4. Sketch showing the three main processes (parameteriza-
tions) captured by the debris-cover extent and thickness evolution
module. γz,t = γz−1,t + γz+1,t
2
,
if γz,t−1 = 0
and z < ELA and dELAt−9,t
dt
> 0. (5) (5) the debris fraction remains stable. Such a neutral up to posi-
tive evolution has for example been observed in the Karako-
ram over the last 40 years (Herreid et al., 2015). Since there
are not enough observational data that would allow for con-
straining the parameter, the time window of 10 years, which
accounts for the time it takes for the debris cover of a glacier
to respond to changes in the glacier-wide mass balance, is
based on judgement. Our implementation also accounts for
the observation that in elevations with limited debris, rela-
tive expansion is slower compared to elevations with a high
debris fraction. Small debris fractions are often associated
with small moraines or isolated debris patches, indicative of
relatively limited debris concentration in the ice. Areas with
abundant debris cover may grow faster due to enhanced de-
bris supply from melt-out or due to ice-flow changes (Ander-
son, 2000; Anderson et al., 2021b). Note that this equation is This process is discretized within elevation bands of 10 m. The number of elevation bands h without debris at time t −1
that can gain debris from a nearby elevation band at time t
(we use yearly time steps) applying Eq. (5) is equal to the
rise in the ELA over the last 10 years, determined using linear
regression of the values (ELA(t−9,t)), i.e. #h = dELAt−9,t
dt · 10
. (6) #h = dELAt−9,t
dt · 10
. (6) In other words, the up-glacier expansion of debris cover
rises with the ELA rise. The thickness of new debris is arbi-
trarily set to 1 cm. If the slope of the above linear regression
is zero or negative, Eq. (5) is not applied. L. Compagno et al.: Regional-scale modelling of supraglacial debris L. Compagno et al.: Regional-scale modelling of supraglacial debris Figure 3. Schematic of the melt enhancement factor g (dimensionless) as a function of debris thickness for three different glaciers (green
lines; k is the value of kdebris calibrated for each glacier). The coloured, dashed boxes show regions in which the different cases of Eqs. (2)
and (3) apply. MB: mass balance. only applied in the glacier ablation zone. Debris cover is not
permitted in the accumulation zone. Figure 4. Sketch showing the three main processes (parameteriza-
tions) captured by the debris-cover extent and thickness evolution
module. 3.2
Debris-cover evolution Note that ponds and ice cliffs, known to
influence the surface mass balance of debris-covered glaciers
as well (Ragettli et al., 2016; Miles et al., 2018; Rounce et al.,
2018), are implicitly accounted for during the Østrem curve
fitting procedure (see Sect. 2.2), since their effect is already
accounted for in the mass-balance data (Miles et al., 2021). We do not model ponds and ice cliffs explicitly because (1) of
the lack of detailed information that would be needed for
accurate calibration and evaluation at the regional scale and
(2) their long-term and future evolution is uncertain (Narama
et al., 2017; Watson et al., 2017; Chand and Watanabe, 2019;
Mölg et al., 2020) and requires small-scale, specific process
models to be captured (e.g. Buri et al., 2021; Kneib et al.,
2021). We also consider debris expansion to be inversely propor-
tional to the 10-year moving average of glacier-wide mass
balance B(t−9,t). By doing so, we parametrize ice-dynamical
processes: in the case of negative long-term mass balance,
the debris-cover fraction increases, resulting in an accumula-
tion of debris; in the case of positive long-term mass balance,
the debris fraction decreases, mimicking debris evacuation
by ice flow (Anderson and Anderson, 2016; Ferguson and
Vieli, 2021); and in case of neutral long-term mass balance, The Cryosphere, 16, 1697–1718, 2022 https://doi.org/10.5194/tc-16-1697-2022 L. Compagno et al.: Regional-scale modelling of supraglacial debris
1703
Figure 3. Schematic of the melt enhancement factor g (dimensionless) as a function of debris thickness for three different glaciers (green
lines; k is the value of kdebris calibrated for each glacier). The coloured, dashed boxes show regions in which the different cases of Eqs. (2)
and (3) apply. MB: mass balance. 1703 4.1
Mass balance 2021)) and to decreasing
debris thickness for positive mass balances (thus mimicking
the evacuation of debris with enhanced flow). This is in line
with the few direct observations that are available (e.g. Gib-
son et al., 2017; Verhaegen et al., 2020). h0 is the mean debris
thickness of the glacier at the inventory year. It parameterizes
the effect that glaciers with a low mean debris thickness will
thicken slower compared to glaciers with a high mean debris
thickness. This is motivated by the assumption that glaciers
with thick debris are likely to have a higher englacial debris
concentration, indicative of high debris supplies from the sur-
roundings. A glacier-specific, three-step calibration procedure is used
to account for the sensitivity of each glacier to the local
climate (as used in Huss and Hock, 2015). The goal is to
match the glacier-specific mass balance between 2000 and
2019 provided by Hugonnet et al. (2021). The accepted misfit
is 0.01 m w.e. a−1. First, the precipitation given by the forc-
ing data set is adjusted with a multiplicative enhancement
factor that is allowed to vary between 0.6 and 2.0. Second,
the degree-day factors are varied in a range of between 1.75
and 4.5 mm d−1 K for DDFsnow, and DDFice is prescribed to
always relate to a factor of 2 to DDFsnow. If the second step is
not needed, the default values of 3 and 6 mm d−1 K are used
for DDFsnow and DDFice, respectively. Third, the local air
temperature is adjusted. The steps are applied sequentially,
meaning that the calibration is considered to be completed
as soon as the observations are matched within the tolerated
misfit (see Huss and Hock, 2015, for more details). Steps 2
and 3 may thus not be applied in all cases. Indeed, 44 % of
the glaciers found an agreement in the first step; 30 % found
one in the second step; and 26 % found one in the third step. In order to investigate the importance of the new debris-
cover module when projecting future glacier evolution
(through the paper, this approach is termed “explicitly” ac-
counting for debris cover), we establish a second glacier-
specific parameter set where all parameterizations related to
debris cover are disabled (through the paper, this approach is
termed “implicitly” accounting for debris cover). In this case
all glaciers are regarded as clean-ice glaciers. 4.1
Mass balance As we use ob-
served geodetic mass changes for calibration, however, this
parameter set accounts for the effect of debris cover implic-
itly. We re-calibrate GloGEMflow only by adjusting the DDF 4
Model calibration The interaction between the modules (especially of the cali-
bration) and the general workflow of this study are illustrated
in Fig. 2. First, the mass-balance module is calibrated (see
Sect. 4.1), followed by the calibration of the debris-cover
evolution module (see Sect. 4.2). This procedure is iterated
twice, since debris evolution feeds back to mass balance. Fi-
nally, the ice-flow module is calibrated (see Sect. 4.3), and
the three modules are evaluated independently (Sect. 5). hz,t = hz,t−1 + abs(bz,t) · B(t−9,t) · (−1) · h0
· cthickening,if γz,t−1 = 0 and z < ELA,
(7) (7) 3.2.3
Debris thickness evolution As for the lateral expansion of debris, the evolution of de-
bris thickness is linked to internal debris concentration and
glacier mass balance (e.g. Gibson et al., 2017; Mölg et al.,
2019; Verhaegen et al., 2020), as well as to changes in ice-
flow velocity (e.g. Anderson et al., 2021b). Additionally, ex-
ternal drivers such as rock avalanches may locally control the
debris thickness (Shugar and Clague, 2011; Dunning et al.,
2015; Berthier and Brun, 2019). We parametrize the change
in local debris thickness based on an approach that is struc-
turally similar to the one used for lateral debris expansion: L. Compagno et al.: Regional-scale modelling of supraglacial debris elevation at which debris is encountered will remain stable
or decrease if the glacier mass balance is positive, e.g. due to
negative lateral expansion of debris cover (see Sect. 3.2.1). The procedure was not calibrated due to the need of a con-
siderable amount of accurate data. An evaluation of the per-
formance is found in Sect. 5.2.2. for details). This is done for all glaciers with an area > 2 km2,
with the ice flow being controlled by a deformation-sliding
factor that accounts for both internal ice deformation and
basal sliding. This deformation-sliding factor is calibrated
for each glacier specifically (see Sect. 4.3). For all glaciers
with an area < 2 km2, glacier evolution is modelled with an
elevation-dependent parameterization, which was shown to
be in good agreement with results from higher-order ice-flow
models (Huss et al., 2010). 3.2.2
Up-glacier expansion of debris cover With the above pro-
cess, debris migrates towards higher elevations at the same
rate as the ELA rises. If the ELA does not rise, the maximal https://doi.org/10.5194/tc-16-1697-2022 https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022 The Cryosphere, 16, 1697–1718, 2022 1704 4.1
Mass balance where hz,t is the debris thickness for elevation z and time t;
cthickening is a regional calibration parameter for the debris-
cover thickness evolution. As for lateral debris expansion,
the local mass balance bz,t relates linearly to debris thickness
change. Higher melt rates will lead to faster debris thick-
ening, thus implicitly assuming that debris concentrations
within the ice are homogeneous. Combined with b(z,t), the
long-term glacier-wide mass balance B(t−9,t) implicitly ac-
counts for ice-dynamical processes, e.g. thickening or thin-
ning due to spatial and temporal changes in ice-flow velocity
(see Sect. 7.2 for a detailed discussion). It leads to constant
debris thickness for steady-state conditions (B(t−9,t) = 0) to
a growth of debris thickness with negative mass balances
(thus mimicking dynamic re-distribution of debris and its
compression; Kirkbride (e.g. 2000); Anderson et al. (e.g. 2021b); Ferguson and Vieli (e.g. 2021)) and to decreasing
debris thickness for positive mass balances (thus mimicking
the evacuation of debris with enhanced flow). This is in line
with the few direct observations that are available (e.g. Gib-
son et al., 2017; Verhaegen et al., 2020). h0 is the mean debris
thickness of the glacier at the inventory year. It parameterizes
the effect that glaciers with a low mean debris thickness will
thicken slower compared to glaciers with a high mean debris
thickness. This is motivated by the assumption that glaciers
with thick debris are likely to have a higher englacial debris
concentration, indicative of high debris supplies from the sur-
roundings. where hz,t is the debris thickness for elevation z and time t;
cthickening is a regional calibration parameter for the debris-
cover thickness evolution. As for lateral debris expansion,
the local mass balance bz,t relates linearly to debris thickness
change. Higher melt rates will lead to faster debris thick-
ening, thus implicitly assuming that debris concentrations
within the ice are homogeneous. Combined with b(z,t), the
long-term glacier-wide mass balance B(t−9,t) implicitly ac-
counts for ice-dynamical processes, e.g. thickening or thin-
ning due to spatial and temporal changes in ice-flow velocity
(see Sect. 7.2 for a detailed discussion). It leads to constant
debris thickness for steady-state conditions (B(t−9,t) = 0) to
a growth of debris thickness with negative mass balances
(thus mimicking dynamic re-distribution of debris and its
compression; Kirkbride (e.g. 2000); Anderson et al. (e.g. 2021b); Ferguson and Vieli (e.g. L. Compagno et al.: Regional-scale modelling of supraglacial debris (step 2) but by keeping unaltered the model parameters de-
termined in step 1 (and potentially step 3). This strategy thus
preserves the glacier-specific climate conditions (precipita-
tion totals and temperature) but adjusts the glaciers’ temper-
ature sensitivity for snow and ice melt in order to reproduce
the observed mass change even without directly accounting
for the melt-reduction process of supraglacial debris cover-
age. (step 2) but by keeping unaltered the model parameters de-
termined in step 1 (and potentially step 3). This strategy thus
preserves the glacier-specific climate conditions (precipita-
tion totals and temperature) but adjusts the glaciers’ temper-
ature sensitivity for snow and ice melt in order to reproduce
the observed mass change even without directly accounting
for the melt-reduction process of supraglacial debris cover-
age. 4.2.1
Calibrating lateral debris expansion To determine clateral, we use the debris-cover observations
obtained from the Landsat scenes (set S1, composed of
55 glaciers with debris; see Sect. 2 and e.g. Fig. 5a). First,
the evolution of the debris’ lateral expansion is calculated
for each glacier and each elevation band (e.g. Fig. 5b). Then,
calibration is performed by comparing the lateral expansion
of debris as observed and as modelled using different clateral
factors (ranging from 0 to 5). More specifically, we take the
debris extent detected on each Landsat scene as the initial
condition, and we simulate each glacier independently for
each clateral. Finally, we calculate the root-mean-square er-
ror (RMSE) between modelled and observed lateral debris
expansion over the period captured by our data set and for
each of the 55 glaciers. For each glacier, we select the clateral
which results in the lowest RMSE (see Fig. 5c). The mean
of the selected clateral is clateral = 2.0, while the 0.25 and
0.75 quantiles are clateral = 0.4 and clateral = 4.2, respectively. The mean value is used for all further modelling; i.e. the
same value is applied for all glaciers in HMA, whilst the re-
sult’s sensitivity to the uncertainty in clateral is analysed in
Sect. 5.2.1. Figure 5. (a) Evolution of the debris-covered area of Kangjiaruo
Glacier and Langtang Glacier as inferred from five Landsat scenes. (b) Same as (a) but divided into 10 m elevation bands. The blue line
shows the lateral expansion of the debris cover as observed between
the oldest (1987–1991) and the newest (2015–2019) Landsat scene. (c) Distribution of clateral resulting in the lowest misfit between ob-
served and modelled debris-fraction evolution (given per number
of glaciers). The grey dashed line shows the mean value, while the
grey rectangle shows values within the 0.25 and 0.75 quartiles. 3.3
Ice flow For modelling glacier’s geometry evolution, ice flow is ex-
plicitly accounted for based on the shallow-ice approxima-
tion and the continuity equation (see Zekollari et al., 2019, The Cryosphere, 16, 1697–1718, 2022 https://doi.org/10.5194/tc-16-1697-2022 1705 L. Compagno et al.: Regional-scale modelling of supraglacial debris 5.1
Mass balance To evaluate the performance of the mass-balance module,
we compare the modelled mass balances for 21 glaciers
in HMA against observations provided by the World
Glacier Monitoring Service (WGMS, 2020). For glacier-
wide annual mass balance, the bias (measured–modelled)
is −0.24 m w.e. a−1, and the RMSE is 0.55 m w.e. a−1 (see
Fig. S2). Observations aggregated to elevation bands show a
bias of −0.38 m w.e. a−1 and a RMSE of 0.77 m w.e. a−1. For
glacier-wide winter balance, the bias is 0.23 m w.e. a−1, and
the RMSE is 0.41 m w.e. a−1 (see Fig. S3). These results are
satisfactory in comparison to other regional-scale modelling
studies (e.g. Marzeion et al., 2012; Huss and Hock, 2015;
Radi´c and Hock, 2014). 4.3
Ice flow The ice-flow module is initialized and calibrated by generat-
ing a glacier-specific steady state for a specified point in time
in the past. The exact timing of this point in time depends on
both climate and glacier response time (see Compagno et al.,
2021, for more details). Starting from this steady state (on
average between 1979–1983 and 1990–1996 in this study),
the glacier is transiently modelled up to the glacier inven-
tory date by using ERA5 reanalysis data. To ensure that the
so-modelled glacier volume and length are consistent with
the available observations, the procedure is repeated by itera-
tively changing two parameters: the deformation-sliding fac-
tor and a mass-balance bias applied during the generation of
the steady state (see Zekollari et al., 2019, for more details). Since the entire procedure is performed before the glacier-
specific inventory year, debris cover is considered to be static
(as given by the observations). Once calibrated, the model is
forced by ERA5 reanalysis data (until 2020) and GCM out-
put data to simulate the future glacier evolution (2020 until
2100). 4.2.2
Calibrating debris thickness evolution To determine cthickening, a three-step procedure is used. In
a first step, we map where debris cover appeared for the
first time between ∼1974 (Hexagon satellite imagery) and
∼1989 (oldest Landsat satellite image). This is done for
12 glaciers with Hexagon satellite observations in set S1
(out of the total of 18 glaciers in set S1) within three sub-
regions (Central Himalaya, East Himalaya, and West Tien
Shan). The six remaining glaciers are not used because a
clear signal of debris formation is lacking between ∼1974
and ∼1989. In a second step, we extract the debris thickness
at the locations used in the debris thickness data set of Mc-
Carthy et al. (2021). Recall that the latter data set represents
the debris condition for 2000–2016. Combined, this informa-
tion provides us with an estimate for the mean debris thick-
ening rate between ∼1981 (mean between 1974 and 1989)
and ∼2008 (mean between 2000 and 2016). In a third step,
we compute the difference between observed and modelled
debris-thickening rate for each glacier. To do so, the 12 se- Figure 5. (a) Evolution of the debris-covered area of Kangjiaruo
Glacier and Langtang Glacier as inferred from five Landsat scenes. (b) Same as (a) but divided into 10 m elevation bands. The blue line
shows the lateral expansion of the debris cover as observed between
the oldest (1987–1991) and the newest (2015–2019) Landsat scene. (c) Distribution of clateral resulting in the lowest misfit between ob-
served and modelled debris-fraction evolution (given per number
of glaciers). The grey dashed line shows the mean value, while the
grey rectangle shows values within the 0.25 and 0.75 quartiles. Figure 5. (a) Evolution of the debris-covered area of Kangjiaruo
Glacier and Langtang Glacier as inferred from five Landsat scenes. Figure 5. (a) Evolution of the debris-covered area of Kangjiaruo
Glacier and Langtang Glacier as inferred from five Landsat scenes. (b) Same as (a) but divided into 10 m elevation bands. The blue line
shows the lateral expansion of the debris cover as observed between
the oldest (1987–1991) and the newest (2015–2019) Landsat scene. (c) Distribution of clateral resulting in the lowest misfit between ob-
served and modelled debris-fraction evolution (given per number
of glaciers). The grey dashed line shows the mean value, while the
grey rectangle shows values within the 0.25 and 0.75 quartiles. https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022 1706 L. 4.2.2
Calibrating debris thickness evolution Compagno et al.: Regional-scale modelling of supraglacial debris 1706
L. Compagno et al.: Regional-scale modelling of supraglacial debris
Figure 6. Difference between observed and modelled debris thickness for the period ∼1981–2008 (circles) using different thickness tuning
factor (cthickening) values. Glaciers IDs refer to RGI 6.0 (RGI Consortium, 2017). Figure 6. Difference between observed and modelled debris thickness for the period ∼1981–2008 (circles) using different thickness tuning
factor (cthickening) values. Glaciers IDs refer to RGI 6.0 (RGI Consortium, 2017). lected glaciers are modelled with cthickening values ranging
between 0 and 2 (Fig. 6), and the value minimizing the differ-
ence to observations is chosen. We find cthickening = 1.0. This
value and the result’s sensitivity are evaluated in Sect. 5.2.2. The Cryosphere, 16, 1697–1718, 2022 5.2
Debris evolution In this section, the three parameterizations included in our
debris-evolution module (see Fig. 4) are evaluated against in-
dependent data sets. The Cryosphere, 16, 1697–1718, 2022 https://doi.org/10.5194/tc-16-1697-2022 L. Compagno et al.: Regional-scale modelling of supraglacial debris L. Compagno et al.: Regional-scale modelling of supraglacial debris 1707 Figure 7. (a) Histograms of the misfit between observed and modelled lateral debris expansion when using clateral = 2 (grey) and when
deactivating the module (clateral = 0, orange; no evo: without evolution). (b) Same as (a) but divided into glaciers and distinguishing between
different reference years (corresponding to the Landsat scenes). The colour of each circle represents the misfit (%). (c, d) Modelled debris-
area evolution (red–white–blue shades) and debris-area evolution as observed in the Landsat scenes (black dashed line) for four selected
glaciers (indicated by arrows). Figure 7. (a) Histograms of the misfit between observed and modelled lateral debris expansion when using clateral = 2 (grey) and when
deactivating the module (clateral = 0, orange; no evo: without evolution). (b) Same as (a) but divided into glaciers and distinguishing between
different reference years (corresponding to the Landsat scenes). The colour of each circle represents the misfit (%). (c, d) Modelled debris-
area evolution (red–white–blue shades) and debris-area evolution as observed in the Landsat scenes (black dashed line) for four selected
glaciers (indicated by arrows). 5.2.1
Evaluating lateral expansion of debris ∼1974. In this case, the mean misfit between observed and
modelled debris fraction is 4.4 % (Fig. 7a, orange histogram),
i.e. substantially higher than for the case in which the debris
area evolution is included. The experiment thus shows the
importance of accounting for debris expansion when mod-
elling long-term glacier evolution. To evaluate the parametrization for lateral debris expan-
sion, 18 glaciers (set S1 with Hexagon satellite observations)
within three subregions (Central Himalaya, West Himalaya,
and West Tien Shan) are simulated from 1974 to 2020 (forc-
ing the model with ERA5 climate). The model is initialized
with debris extents extracted from Hexagon satellite images
of ∼1974 (see Sect. 2). The modelled debris-area evolution
is evaluated between 1989 and 2017 against the time series of
debris extents obtained from Landsat (three to five observa-
tions per glacier). We calculate the misfit between modelled
and observed debris-cover fraction for each elevation band. Note that this is the same procedure as used for calibration
(see Sect. 4.2), with the difference that for model initializa-
tion we use debris extents from Hexagon imagery rather than
Landsat (Fig. 7). The mean misfit of debris fraction obtained
by this procedure is 0.7 % (Fig. 7a, grey histogram), with 42
of the 78 evaluations indicating a misfit < ±2 % (Fig. 7a, b). A good model performance is also shown by analysing the
glacier-specific lateral expansion of debris (Fig. 7c, d). 6.1
Glacier-specific simulations In order to illustrate the detailed model results at the scale
of an individual glacier, we focus on the well-investigated
Langtang Glacier (Central Himalaya). A similar illustra-
tion for Baltoro Glacier (Karakoram) and Inylchek Glacier
(West Tien Shan), which showed patterns similar to Lang-
tang Glacier, is given in Figs. S5 and S6, respectively. Figure 8. Misfit between observed and modelled highest elevation
with a lateral debris expansion ≥10 %, 20 %, 30 %, 40 %, and 50 %. The debris-cover evolution module is activated in panel (a) and de-
activated in panel (b). g
g
g
p
y
Figure 9a shows a profile view of the glacier and debris-
cover evolution of Langtang Glacier according to our model
results and SSP245. Under this scenario, Langtang Glacier
would lose 43 % of its 2020 ice volume by 2050, despite the
terminus retreating by less than 300 m (i.e. only 2 % of its
2020 length). The limited retreat can be attributed to the 0.5–
1 m thick insulating debris cover present on the entire glacier
tongue, which reduces Langtang Glaciers’s ice melt by a fac-
tor of about 3 compared to the hypothetical situation with no
debris. The approximately linear dependence between sur-
face mass balance and elevation, which is typical of clean-
ice glaciers, is suppressed for Langtang Glacier, since debris
cover is thicker at lower elevations than it is for higher ones
(Bisset et al., 2020; Miles et al., 2021). This leads to a nearly
homogeneous downwasting of the ablation zone (e.g. Pel-
licciotti et al., 2015; Ragettli et al., 2016) rather than to a
retreat of the terminus (e.g. Benn et al., 2012). This causal-
ity is confirmed when the evolution of Langtang Glacier is
re-computed using the same climatic conditions but when
re-calibrating the model parameters to match the observed
volume changes without activating the debris-cover module
(Fig. 9b). In this case, the glacier would lose 45 % of its 2020
ice volume by 2050 (i.e. very similar to explicit debris mod-
elling) but would retreat by about 2700 m (i.e. 20 % of its
2020 length). The latter is 10 times more than when includ-
ing the effect of supraglacial debris. Figure 8. Misfit between observed and modelled highest elevation
with a lateral debris expansion ≥10 %, 20 %, 30 %, 40 %, and 50 %. L. Compagno et al.: Regional-scale modelling of supraglacial debris L. Compagno et al.: Regional-scale modelling of supraglacial debris 1708 Figure 8. Misfit between observed and modelled highest elevation
with a lateral debris expansion ≥10 %, 20 %, 30 %, 40 %, and 50 %. The debris-cover evolution module is activated in panel (a) and de-
activated in panel (b). indicate high glacier-to-glacier variance of cthickening but also
show that the proposed parametrization is rather insensi-
tive to the weakly constrained value of cthickening (see also
Sect. 7). 6.1
Glacier-specific simulations The debris-cover evolution module is activated in panel (a) and de-
activated in panel (b). one inferred for ∼1974. For this case without up-glacier de-
bris expansion, we re-compute the misfit between observed
and modelled elevation with a debris fraction ≥10 %, 20 %,
30 %, 40 %, and 50 %. This results in a misfit of +55 m,
i.e. almost 2 times larger compared to when the up-glacier
debris expansion parametrization is activated, indicating the
importance of taking up-glacier debris expansion into ac-
count as well. Figure 9a and c show a spatial representation of the debris-
cover evolution according to the three implemented param-
eterizations. At 5500 m a.s.l. for instance, the fraction of de-
bris increased by 87 % between 2025 and 2100. This result is
driven by the projected lateral debris expansion. As a result
of the projected up-glacier migration, instead, the maximum
elevation with supraglacial debris would increase by 280 m
for the same time period. Finally, the local debris thickness
at 5500 m a.s.l. is projected to increase by 0.23 m over the
period 2025–2100. 5.2.2
Evaluating debris up-glacier expansion To evaluate the parametrization for the up-glacier debris
expansion, we use the same experiment setup as above
(Sect. 5.2.1). We initialize the model in 1974 and force it un-
til 2020 but now focus on the transition zone between debris-
covered and bare-ice surfaces. For each glacier with observed
debris extents from Hexagon and Landsat (of set S1), we
extract from the Landsat scenes the highest elevation bands
that have a debris-covered fraction of ≥10 %, 20 %, 30 %,
40 %, and 50 %. We perform the same extraction procedure
for the modelling results. Then we compute for each glacier
and each Landsat scene the elevation misfit between obser-
vations and modelling results (Fig. 8). The mean misfit is of
+27 m. The performance of our parametrization for lateral debris
expansion can also be evaluated by disabling this process in
the model. We do so by initializing the model as above but
by prescribing a constant debris cover, taken to be the one of Again, an alternative way for evaluating our approach is to
turn off the up-glacier expansion parametrization, thus pre-
scribing a temporally constant debris cover, set to be the https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022 L. Compagno et al.: Regional-scale modelling of supraglacial debris L. Compagno et al.: Regional-scale modelling of supraglacial debris 1709 Figure 9. (a) Modelled evolution of Langtang Glacier when debris is explicitly accounted for. The results refer to SSP245. Note t
debris thickness (grey) is exaggerated by a factor of 500 for visibility. The three parameterizations included in the debris-cover
(cf. Sect. 3.2 and Fig. 4) are indicated by the circled, coloured numbers and described in the text. (b) Same as (a) but accounting for
implicitly; i.e. glacier evolution is not modelled with the new debris-cover module but by re-calibrating some of the model param
match observed long-term mass balance (see Sect. 4.1 for details). (c) Model results extrapolated to 2D (see Supplement for the
used for extrapolating from one to two dimensions, and note that the extrapolation is for visualization purposes only; i.e. it does no
the presented results). For every SSP, the evolution of (d) debris-cover fraction, (e) glacier volume with explicit debris-cover mo
(g) debris thickness, and (h) glacier area with explicit modelling is shown. (f, i) For every SSP the difference in glacier volume a
obtained when explicitly and implicitly modelling the debris cover. The shaded ranges represent 1 standard deviation of all climate
members included in a given SSP. Figure 9. (a) Modelled evolution of Langtang Glacier when debris is explicitly accounted for. The results refer to SSP245. Note that the
debris thickness (grey) is exaggerated by a factor of 500 for visibility. The three parameterizations included in the debris-cover module
(cf. Sect. 3.2 and Fig. 4) are indicated by the circled, coloured numbers and described in the text. (b) Same as (a) but accounting for debris
implicitly; i.e. glacier evolution is not modelled with the new debris-cover module but by re-calibrating some of the model parameters to
match observed long-term mass balance (see Sect. 4.1 for details). (c) Model results extrapolated to 2D (see Supplement for the method
used for extrapolating from one to two dimensions, and note that the extrapolation is for visualization purposes only; i.e. it does not affect
the presented results). For every SSP, the evolution of (d) debris-cover fraction, (e) glacier volume with explicit debris-cover modelling,
(g) debris thickness, and (h) glacier area with explicit modelling is shown. (f, i) For every SSP the difference in glacier volume and area
obtained when explicitly and implicitly modelling the debris cover. L. Compagno et al.: Regional-scale modelling of supraglacial debris The shaded ranges represent 1 standard deviation of all climate model
members included in a given SSP. simulations show that the fraction of debris-covered area
is expected to increase until 2060, reaching a maximum of
55±2 % (mean and standard deviation of all model members
considering SSP245) relative to the remaining glacier area. After reaching this debris peak fraction, a fast decline is mod-
elled due to disintegration of the debris-covered tongue. De- debris-covered area and debris thickness increase, thus re-
ducing ice melt. Nevertheless, higher mass loss also leads to
glacier retreat and downwasting of the debris-covered glacier
tongue, thus reducing debris-cover extent due to glacier area
loss. Therefore, a competition between debris increase and
reduction arises after 2060. For Langtang Glacier, model 5.2.3
Evaluating debris thickness evolution To evaluate the debris thickness evolution parametrization,
we compare model results against the evaluation data set S2,
i.e. 11 glaciers distributed in four regions (East Himalaya,
Karakoram, West Tien Shan, and Inner Tibet). By setting
cthickening = 1.0, the mean misfit between observed and mod-
elled debris-thickening rate is 0.07 m. For cthickening = 0.0
(i.e. no debris thickness evolution), it is 0.17 m (see Fig. S4). The model, thus, performs better when the debris thickness
evolution parametrization is activated. Taken together, this
evaluation and the calibration results (Sect. 4.2.2) not only The presence of supraglacial debris alters glacier mass
balance, hence influencing the debris-cover evolution itself. With higher mass loss, our model results show that both The Cryosphere, 16, 1697–1718, 2022 https://doi.org/10.5194/tc-16-1697-2022 Compagno et al.: Regional-scale modelling of supraglacial debris 6.2
Regional glacier evolution (
)
(
)
Compared to 2020, the modelled mean debris thickness
of Langtang Glacier is expected to increase by 35 ± 5 % and
reach its maximum in 2065 (SSP245). The variation between
individual SSPs is relatively small. Different SSPs give rise
to different thickness evolution trajectories however, reach-
ing between −7±10 % (SSP119) and −75±19 % (SSP585)
of the 2020 debris thickness by 2100. This counter-intuitive
decrease in average debris thickness can be explained by the
evolution of both debris extent as well as glacier geometry. Indeed, the expansion of thin debris to higher areas and the
loss of presently thick debris on the downwasting tongue re-
sults in a mean debris thickness decrease (see also Sect. 7). When neglecting the evolution of debris extent and thickness
(i.e. the change in debris thickness is only due to glacier ge-
ometry change), the mean debris thickness would decrease
by between 30 ± 18 % (SSP119) and 80 ± 35 % (SSP585)
by 2100. Together with the debris-fraction evolution, this
demonstrates that it is relevant to account for transient debris-
cover changes in process-based models, at least for low- to
medium-emission scenarios (Fig. 9d). The area-averaged debris-cover fraction of all glaciers in
HMA is expected to be between 14 % and 24 % by 2100
compared to the 12 %–13 % observed today (Fig. 10a; note
that these numbers refer to glaciers with an area > 2 km2
and that the debris-cover fractions are computed for the tran-
siently evolving glacier areas). Generally, the debris-cover
fraction is projected to be higher for higher-emission sce-
narios (i.e. scenarios implying a higher air temperature in-
crease). The expected increase in debris-cover fraction is
due to both lateral and up-glacier expansion. Without ac-
counting for debris-cover evolution, the debris-cover frac-
tion, however, is projected to be between 8 ± 1 % (SSP119)
and 6 ± 1 % (SSP585). This highlights the importance of
accounting for dynamic debris-cover evolution in process-
based studies. Our results also show that under low-emission
scenarios, the competing processes of debris expansion and
glacier retreat tend to reach an equilibrium at the end of the
century. For high-emission scenarios, instead, debris-cover
expansion dominates over glacier retreat. By 2100 and when explicitly modelling debris-cover
changes, Langtang Glacier is projected to lose between 69 ±
14 % (SSP119) and 98±2 % (SSP585) of its 2020 ice volume
(Fig. 9e). https://doi.org/10.5194/tc-16-1697-2022 https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022 1710
L. Compagno et al.: Regional-scale modelling of supraglacial debris
Figure 10. Evolution of (a) debris-covered fraction and (b) debris thickness for all HMA glaciers and as an average for the respective SSP. The shaded bands represent 1 standard deviation of all climate model members. The grey line indicates the case in which the debris-evolution
module is disabled and only today’s debris cover is used in the modelling. L. Compagno et al.: Regional-scale modelling of supraglacial debris L. Compagno et al.: Regional-scale modelling of supraglacial debr 1710 pagno et al.: Regional-scale modelling of supraglacial debris
k
f
ll
l
i
d
f
h
i
SS Figure 10. Evolution of (a) debris-covered fraction and (b) debris thickness for all HMA glaciers and as an average for the respective SSP. The shaded bands represent 1 standard deviation of all climate model members. The grey line indicates the case in which the debris-evolution
module is disabled and only today’s debris cover is used in the modelling. volume loss only differing by between 1 % and 6 %, depend-
ing on the SSP (Fig. 9h, i). This indicates that constraining
the model to past glacier mass loss yields similar results, even
when considering Langtang Glacier to be a clean-ice glacier. In that case, however, surface mass-balance gradients would
not consider the effect of debris cover (see e.g. Fig. S7),
with consequences for the geometry evolution, runoff, and
surface-elevation feedbacks. pending on the emission scenario, the debris-covered fraction
reaches between 28 ± 6 % (SSP119) and 18 ± 5 % (SSP585)
by 2100 (Fig. 9f). Note that the fraction refers to the evolving
glacier geometry and not to the geometry at inventory date. By turning off the debris-evolution parametrization (expan-
sion and thickening) and by using presently observed debris
extent and thickness instead (grey line in Fig. 9f), the de-
bris fraction would continuously decrease, reaching between
14 ± 7 % (SSP119) and 1 ± 1 % in 2100 (SSP585). L. Compagno et al.: Regional-scale modelling of supraglacial debris high-emission scenarios. Locally, however, much higher de-
bris thickness increases are modelled, but these are offset at
many places by the overall reduction in glacier area. Indeed,
the small overall change is explained by (1) the disintegra-
tion of glacier tongues, where the debris is generally thick-
est; (2) lateral debris expansion, which is most efficient at in-
termediate elevations with relatively thin debris; and (3) up-
glacier expansion of debris, which forms new thin debris. If
debris evolution is not modelled (i.e. static debris), the mod-
elled overall change in debris thickness would be more pro-
nounced, with an average of between −33 ± 10 % (SSP119)
and −66 ± 5 % (SSP585) for the period 2020–2100. fer when not explicitly accounting for debris cover (see
Fig. S7). Even though for all glaciers the mass change be-
tween 2000 and 2019 is constrained to match the same data
(Hugonnet et al., 2021), the spatial mass-balance distribution
influences the geometry evolution and, hence, mass turnover
and surface flow velocity of glaciers. Aggregated over all of HMA and considered in terms
of glacier volume and area changes, the difference in
the results between explicitly and implicitly modelling de-
bris cover is relatively small. On average, DDFice is of
3.49 mm d−1 ◦C−1 when the debris cover is modelled ex-
plicitly and of 3.55 mm d−1 ◦C−1 when it is modelled im-
plicitly. However, on the single-glacier scale, differences in
DDFice are larger (e.g. 0.47 mm d−1 ◦C−1 for Langtang and
0.20 mm d−1 ◦C−1 for Inylchek). This, however, does not
mean that debris can be neglected in climate impact stud-
ies. In fact, accounting for the debris cover explicitly enables
the model to mimic the driving processes, rather than com-
pensating the lack of model capabilities through a suitable
parameter choice. This is important, especially when results
other than area and volume changes are of interest. Indeed,
quantities such as the local mass balance, the glaciers’ ice-
flow velocity and mass turnover, the glacier’s length change,
and water runoff are different when explicitly accounting for
supraglacial debris and its temporal evolution. These quanti-
ties, in turn, have to be modelled appropriately when aiming
at anticipating other glacier-related processes, such as haz-
ards from ice-dammed or proglacial lakes or potential slope
instabilities. 7.1
Importance of accounting for debris-cover
evolution We demonstrated the difference between explicitly and im-
plicitly accounting for the effect of debris cover, as well as
the importance of modelling debris-cover evolution. The goal
was to assess whether such processes need to be taken into
account when modelling glacier evolution at the local to re-
gional/global scale. The most significant differences emerge
for computed glacier length changes, with further differ-
ences being found for volume and area evolution, especially
for glaciers with high debris-cover fraction and thickness at
present (see the example of Langtang Glacier; Fig. 9a, b). Moreover, modelled surface mass-balance gradients also dif- L. Compagno et al.: Regional-scale modelling of supraglacial debris By 2100, HMA glaciers are expected to lose between
35±15 % (SSP119) and 80±11 % (SSP585) of their 2020 ice
volume when debris cover is explicitly modelled (Fig. 11a). By modelling debris cover implicitly (i.e. using the same cli-
matic conditions together with parameters re-calibrated to
match observed volume change but without activating the
debris-cover module), the simulated mean ice volume loss
would be only between 1 % (SSP119) and 3 % (SSP585)
higher (Fig. S8a). The difference is small because (1) only
about 12 %–13 % of the glacier area in HMA is currently
debris-covered (Herreid and Pellicciotti, 2020); (2) the mod-
els are constrained to reproduce observed mass change, both
when accounting and neglecting the effects of debris cover;
and (3) both positive and negative differences can be found
at the level of individual glaciers, caused by different debris
and geometry evolution of each glacier. By dividing glaciers
into classes of debris-cover fraction, the difference between
modelling debris cover explicitly or implicitly becomes more
evident, reaching up to 30 % of the difference in volume for
glaciers with a present debris fraction > 0.5 (see Fig. 11b). This shows that the difference driven by non-linear feedback
of debris cover on mass balance becomes relevant for glaciers
with extensive debris-cover fractions. For differences in both
future area evolution and spatially distributed glacier evolu-
tion, see Figs. S9 and S10. Compared to the static representation of supraglacial
debris cover that is presently included in some regional
to global glacier models (e.g. Kraaijenbrink et al., 2017;
Rounce et al., 2021), the expected increase in both debris-
cover fraction and local debris thickness will enhance the in-
sulating effects of the debris cover. Figure 10a and b show
that if the debris area and thickness would not evolve through
time, the future debris-cover fraction and mean debris thick-
ness would significantly decrease. This is related to the loss
of frontal area projected in such a case, since the frontal area
typically features the highest concentration of supraglacial
debris. However, a significant decrease in debris cover has
been neither observed over the past decades (Stokes et al.,
2007; Bhambri et al., 2011; Bolch et al., 2011; Shukla and
Qadir, 2016; Tielidze et al., 2020) nor modelled in glacier-
specific studies specifically addressing the future evolution
of debris (Jouvet et al., 2011; Rowan et al., 2015; Kienholz
et al., 2017; Verhaegen et al., 2020). 6.2
Regional glacier evolution If debris cover is implicitly taken into account, very
similar results are obtained, with simulated 2100 area and Between 2020 and 2100, the area-averaged debris-cover
thickness of all glaciers in HMA (again, with area > 2 km2
and not of surge type) is expected to slightly increase by
about 5 % or 2 cm compared to today; see Fig. 10b. Inter-
estingly, a very similar change is found for both low- and https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022 1711 L. Compagno et al.: Regional-scale modelling of supraglacial debris https://doi.org/10.5194/tc-16-1697-2022 7.3
Velocity However, volume differences for individual large
and strongly debris-covered glaciers can be as high as 18 %
when the debris-evolution module is disabled (e.g. Langtang
Glacier at 2 %, Baltoro Glacier at 8 %, and Inylchek at 1 %). To assess the effect of explicitly accounting for debris-
cover dynamics on the modelled mass turnover, we com-
pare the computed surface velocities against NASA’s MEa-
SUREs (Making Earth Science Data Records for Use in Re-
search Environments) ITS_LIVE (Inter-mission Time Series
of Land Ice Velocity and Elevation) surface velocity data
set (Gardner et al., 2019). The comparison is performed for
all glaciers in HMA with a debris-cover fraction > 0.3. For
ITS_LIVE, we use the 120 m resolution composite data cov-
ering the period 1985–2018. To account for spatial variations
in surface velocity, we compare modelled and observed sur-
face velocities aggregated to 100 m elevation bands at the
scale of each individual glacier. We exclude the glaciers’ ac-
cumulation area, since the insufficient contrast in optical im-
ages impedes feature tracking, thus resulting in higher uncer-
tainties in the ITS_LIVE velocity. About two-thirds of the 3767 elevation bands investigated
with this selection show a velocity that is closer to observa-
tions when the debris cover is explicitly accounted for. The
mean modelled velocity is 16 % slower than in the case in
which the debris cover is neglected (Fig. S11). This indi-
cates that the mass turnover is indeed smaller when debris
cover is accounted for, which is consistent with the avail-
able ITS_LIVE observations. The smaller ice velocities are
in line with findings of more theoretical and process-based
modelling studies on the dynamics of debris-covered glaciers
(Anderson and Anderson, 2016; Ferguson and Vieli, 2021). Additional uncertainties arise also from the parameteriza-
tions themselves (Eqs. 4, 5, and 7). In a simplified but real-
istic way, our approach parameterizes the evolution of debris
cover on glaciers, and it is based on debris-evolution patterns
observed in the last decades. We acknowledge that this is not
the only way that debris evolution could be parameterized. Explicitly accounting for the dynamics of debris re-
distribution (as opposed to implicitly; see Sect. 3.2.3) could
be an alternative option. Indeed, where the ice-flow veloc-
ity decreases, the local debris thickness increases and vice
versa, due to the convergence of debris particles. Such an
approach was followed by Anderson et al. (2021b), for ex-
ample. 7.2
Model sensitivity and uncertainties Uncertainties in glacier and debris evolution can arise from
many factors. In Zekollari et al. (2019) and Compagno et al. (2021), limited sensitivity to the initial ice thickness distribu-
tion, the geometry-initialization method, the model param-
eters, and the data used for mass-balance calibration were https://doi.org/10.5194/tc-16-1697-2022 https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022 L. Compagno et al.: Regional-scale modelling of supraglacial debris L. Compagno et al.: Regional-scale modelling of supraglacial debris 1712 Figure 11. (a) Evolution of the glacier volume for all glaciers in HMA when explicitly modelling debris-cover changes. Results are aggre-
gated to the five SSPs. (b) Difference in volume (mean over 2090–2100) between implicit and explicit debris-cover modelling classified for
different debris-cover fractions. The number of modelled glaciers per class is given in grey. The shaded bands represent 1 standard deviation
of all climate model members for a given SSP. Figure 11. (a) Evolution of the glacier volume for all glaciers in HMA when explicitly modelling debris-cover changes. Results are aggre-
gated to the five SSPs. (b) Difference in volume (mean over 2090–2100) between implicit and explicit debris-cover modelling classified for
different debris-cover fractions. The number of modelled glaciers per class is given in grey. The shaded bands represent 1 standard deviation
of all climate model members for a given SSP. 7.3
Velocity We decided not to include such effects explicitly be-
cause of (1) the absence of data to calibrate and validate a
more complex parameterization at regional scales and (2) the
small sensitivity of regional-scale glacier volume and area to
changes in debris cover (see Sect. 7.1). 7.3
Velocity found. In this study, additional uncertainties arise from the
parameters clateral and cthickening of the debris-evolution mod-
ule, i.e. from the lateral debris expansion and thickening pa-
rameters. To test model sensitivity to variations in these fac-
tors, we re-compute the future evolution of all glaciers with
clateral = 1.0 and clateral = 3.0 (compared to the reference
value clateral = 2.0) and with cthickening = 0.5 and cthickening =
1.5 (reference: cthickening = 1.0). The results indicate that
these variations in both factors have little impact on the ice
volume loss modelled for 2020–2100, with a difference of
less than ±1 %. By disabling the debris-evolution module
(i.e. with clateral = 0 and cthickening = 0), the regional ice vol-
ume loss would be about 1 % higher. This small difference is
again due to the fact that about 87 %–88 % of glacier area is
debris-free. However, volume differences for individual large
and strongly debris-covered glaciers can be as high as 18 %
when the debris-evolution module is disabled (e.g. Langtang
Glacier at 2 %, Baltoro Glacier at 8 %, and Inylchek at 1 %). Additional uncertainties arise also from the parameteriza-
tions themselves (Eqs. 4, 5, and 7). In a simplified but real-
istic way, our approach parameterizes the evolution of debris
cover on glaciers, and it is based on debris-evolution patterns
observed in the last decades. We acknowledge that this is not
the only way that debris evolution could be parameterized. found. In this study, additional uncertainties arise from the
parameters clateral and cthickening of the debris-evolution mod-
ule, i.e. from the lateral debris expansion and thickening pa-
rameters. To test model sensitivity to variations in these fac-
tors, we re-compute the future evolution of all glaciers with
clateral = 1.0 and clateral = 3.0 (compared to the reference
value clateral = 2.0) and with cthickening = 0.5 and cthickening =
1.5 (reference: cthickening = 1.0). The results indicate that
these variations in both factors have little impact on the ice
volume loss modelled for 2020–2100, with a difference of
less than ±1 %. By disabling the debris-evolution module
(i.e. with clateral = 0 and cthickening = 0), the regional ice vol-
ume loss would be about 1 % higher. This small difference is
again due to the fact that about 87 %–88 % of glacier area is
debris-free. L. Compagno et al.: Regional-scale modelling of supraglacial debris 1713 L. Compagno et al.: Regional-scale modelling of supraglacial debris L. Compagno et al.: Regional-scale modelling of supraglacial debris Figure 12. Comparison of modelled volume changes for HMA with values from Marzeion et al. (2020) and Edwards et al. (2021). Changes
are expressed with respect to the 2020 baseline. From left to right, the abbreviations (GLIMB, GLacIer energy Mass Balance; GloGEM,
Global Glacier Evolution Model; JULES, Joint UK Land Environment Simulator; KRA2017; MAR2012; OGGM, Open Global Glacier
Model; PyGEM, Python Glacier Evolution Model; RAD2014; and WAL2001) stand for Sakai and Fujita (2017), Huss and Hock (2015),
Shannon et al. (2019), Kraaijenbrink et al. (2017), Marzeion et al. (2012), Maussion et al. (2019), Rounce et al. (2020), Radi´c and Hock
(2014), and Van de Wal and Wild (2001). The dashed lines correspond to the mean volume changes of this study. RCP: Representative
Concentration Pathway. Figure 12. Comparison of modelled volume changes for HMA with values from Marzeion et al. (2020) and Edwards et al. (2021). Changes
are expressed with respect to the 2020 baseline. From left to right, the abbreviations (GLIMB, GLacIer energy Mass Balance; GloGEM,
Global Glacier Evolution Model; JULES, Joint UK Land Environment Simulator; KRA2017; MAR2012; OGGM, Open Global Glacier
Model; PyGEM, Python Glacier Evolution Model; RAD2014; and WAL2001) stand for Sakai and Fujita (2017), Huss and Hock (2015),
Shannon et al. (2019), Kraaijenbrink et al. (2017), Marzeion et al. (2012), Maussion et al. (2019), Rounce et al. (2020), Radi´c and Hock
(2014), and Van de Wal and Wild (2001). The dashed lines correspond to the mean volume changes of this study. RCP: Representative
Concentration Pathway. jita, 2017; Maussion et al., 2019; Shannon et al., 2019;
Rounce et al., 2020) that participated in the Glacier Model
Intercomparison Project, phase 2 (GlacierMIP2; Marzeion
et al., 2020). Since models participating in GlacierMIP2 used
CMIP5 GCMs to force the glacier evolution models, we ad-
ditionally compare our results to those of Edwards et al. (2021), who used statistical emulation to convert the glacier
volume evolution projected by Marzeion et al. (2020) from
CMIP5 to CMIP6. The individual GlacierMIP2 models used
various methods for modelling both the evolution of glacier
geometry and glacier mass balance, as well as various spatial
discretizations (refer to Table 1 in Marzeion et al., 2020, for
a summary). L. Compagno et al.: Regional-scale modelling of supraglacial debris our results are between 4 % less negative and 2 % more neg-
ative for SSP126 and SSP585, respectively, than the mean
result of Edwards et al. (2021) using the same climate forc-
ing data (Fig. 12). We suspect that the somewhat larger mass loss compared
to GlacierMIP2 can be attributed to higher climate sensitiv-
ity of the CMIP6 GCMs used in our study compared to the
CMIP5 GCMs (Wyser et al., 2020) used in Marzeion et al. (2020). In general, the explicit inclusion of debris-cover dy-
namics does not result in fundamentally different volume
projections at the regional scale. Our results, however, per-
mit a better representation of the transient processes that
control the changes at the scale of individual glaciers. For
a detailed comparison between our results and various site-
specific studies for HMA, we refer the reader to the Supple-
ment. We also compare our results specifically to Kraaijenbrink
et al. (2017), which is the only regional study available so far
that explicitly accounted for the effect of debris cover. The
study used remote sensing data to determine the spatial dis-
tribution of debris, as well as debris surface-temperature to
estimate debris thickness and its relation to ice melt. Debris-
cover extent and thickness was considered to be static, simi-
larly to the case shown in Fig. 10 (grey line). Also, Kraaijen-
brink et al. (2017) did not simulate ice flow explicitly and did
not calibrate the mass-balance model against glacier-specific
observations. Our results for HMA’s total glacier volume loss
during 2020–2100 are slightly more negative (between 5 %
and 11 % for SSP126 and SSP585, respectively) than the pro-
jections of GlacierMIP2. They also project more mass loss
than Kraaijenbrink et al. (2017), with differences between
17 % and 13 % for SSP126 and SSP585, respectively. Finally, 7.4
Comparison with other studies We compare our HMA-wide results against the ones from
the nine global glacier models (Van de Wal and Wild,
2001; Marzeion et al., 2012; Radi´c and Hock, 2014; Huss
and Hock, 2015; Kraaijenbrink et al., 2017; Sakai and Fu- The Cryosphere, 16, 1697–1718, 2022 https://doi.org/10.5194/tc-16-1697-2022 L. Compagno et al.: Regional-scale modelling of supraglacial debris
1713 L. Compagno et al.: Regional-scale modelling of supraglacial debris ral evolution of both the spatial distribution and the thick-
ness of the supraglacial debris is controlled by the glacier’s
mass balance, equilibrium line altitude, and pre-existing de-
bris properties. The mass-balance module was calibrated
through glacier-specific geodetic ice volume changes, while
the debris-evolution module was calibrated and evaluated in-
dependently with remote sensing observations. The model
was applied to all HMA glaciers with two modalities: one
where the supraglacial debris cover is accounted for explic-
itly and one where this is only done implicitly, i.e. by using
the same climatic conditions but by re-calibrating the param-
eters of the mass-balance module to match observed volume
change whilst pretending all glaciers to be debris-free. sensing observations and field-based methods will be valu-
able in this respect. Data availability. Data is available at https://doi.org/10.3929/ethz-
b-000544323 (Loris et al., 2022). Data availability. Data is available at https://doi.org/10.3929/ethz-
b-000544323 (Loris et al., 2022). Supplement. The supplement related to this article is available on-
line at: https://doi.org/10.5194/tc-16-1697-2022-supplement. Author contributions. LC, DF, MH, and HZ conceived the study. LC performed the numerical modelling with support from MH,
ESM, MJM, HZ, AD, FP, and DF. The original code was devel-
oped by MH (mass-balance and debris-evolution modules) and by
HZ (ice-flow module). MJM and ESM produced the estimates of
debris thickness and the glacier-specific Østrem curves. ESM pro-
duced the debris-cover map from the multiple Landsat satellite im-
ages. AD georeferenced and orthorectified the Hexagon images. LC
wrote the manuscript and produced the figures, with contributions
from all other co-authors. When explicitly modelling debris, we found that both the
debris-covered fraction and the local debris thickness will
increase in the future. This is related to the ongoing atmo-
spheric warming, with larger debris-cover changes projected
for scenarios of higher warming. Averaged over the transient
glacier area, our approach anticipates the mean debris thick-
ness to increase only slightly. This is due to the expansion of
areas where new, thin debris is expected to form. Perhaps surprisingly, explicitly modelling debris-cover
evolution has only a small effect on the regional-scale glacier
volume and area evolution. The difference to the case in
which the debris cover is modelled implicitly is below 3 %. This is due to the fact that the majority of HMA’s glaciers
are debris-free. L. Compagno et al.: Regional-scale modelling of supraglacial debris The regional volume and area evolution is
an average over a vast number of glaciers, with individual
glacier-specific signals cancelling each other out due to site-
specific evolution of debris and geometry for each glacier. This does not mean that the glaciers’ debris cover can be ne-
glected or that it is encouraged to account for debris only
implicitly. At the glacier-specific scale, in fact, the difference
between explicitly and implicitly modelling debris cover be-
comes important. We found, for example, that explicit mod-
elling of debris can significantly decrease the mass-balance
gradient of a given glacier. This results in turn is a reduced
mass turnover, with consequences for the future evolution of
the glacier’s geometry, the modelled surface ice velocities,
or the methods that use considerations of mass turnover for
estimating glacier ice thickness (for overviews, see Farinotti
et al., 2017, 2021). At the level of individual glaciers, such
quantities are important, as they can have implications on
e.g. water availability and natural hazards. Competing interests. At least one of the (co-)authors is a member
of the editorial board of The Cryosphere. The peer-review process
was guided by an independent editor, and the authors also have no
other competing interests to declare. Disclaimer. Publisher’s note: Copernicus Publications remains
neutral with regard to jurisdictional claims in published maps and
institutional affiliations. Acknowledgements. We thank the RGI Consortium for the global
glacier inventory data. We acknowledge the ECMWF for the ERA5
reanalysis and CMIP for the GCM outputs. We also thank the World
Glacier Monitoring Service (WGMS) for providing mass-balance
and length change measurements. Finally, we are grateful to the
editor, Andreas Vieli, and the two reviewers, Ben Marzeion and
Leif S. Anderson, for their numerous constructive remarks and sug-
gestions. Financial support. This
research
has
been
supported
by
the
Schweizerischer
Nationalfonds
zur
Förderung
der
wis-
senschaftlichen Forschung (grant no. 184634), the European
Union’s Horizon 2020 research and innovation programme (grant
agreement no. 869304, PROTECT contribution no. 34; grant
no. 772751, RAVEN), the Horizon 2020 Marie Skłodowska-Curie
Actions (grant no. 799904), and the Fonds de la Recherche Scien-
tifique (FNRS; postdoctoral fellowship, chargé de recherches). Based on the above, we encourage explicitly accounting
for the temporal evolution of supraglacial debris when mod-
elling debris-covered glaciers. 8
Conclusions In this study, we presented a new module for simulating the
spatiotemporal evolution of supraglacial debris. We imple-
mented the new approach into the glacier model GloGEM-
flow and showed its applicability from the single-glacier
to the regional scale. By relying on glacier-specific Østrem
curves – i.e. functions that characterize the relation between
debris-cover thickness and ablation rates – the module ac-
counts for the enhanced and reduced melting caused by de-
bris thinner or thicker than 3–4 cm, respectively. The tempo- https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022 1714 References and Brun, F.: Karakoram geodetic glacier mass balances
between 2008 and 2016: Persistence of the anomaly and influ-
ence of a large rock avalanche on Siachen Glacier, J. Glaciol.,
65, 494–507, https://doi.org/10.1017/jog.2019.32, 2019. Bhambri, R., Bolch, T., Chaujar, R. K., and Kulshreshtha, S. C.:
Glacier changes in the Garhwal Himalaya, India, from 1968
to 2006 based on remote sensing, J. Glaciol., 57, 543–556,
https://doi.org/10.3189/002214311796905604, 2011. Eyring, V., Bony, S., Meehl, G. A., Senior, C. A., Stevens, B.,
Stouffer, R. J., and Taylor, K. E.: Overview of the Coupled
Model Intercomparison Project Phase 6 (CMIP6) experimen-
tal design and organization, Geosci. Model Dev., 9, 1937–1958,
https://doi.org/10.5194/gmd-9-1937-2016, 2016. Biemans, H., Siderius, C., Lutz, A., Nepal, S., Ahmad, B., Hassan,
T., von Bloh, W., Wijngaard, R., Wester, P., Shrestha, A., and
Immerzeel, W. W.: Importance of snow and glacier meltwater
for agriculture on the Indo-Gangetic Plain, Nature Sustainability,
2, 594–601, https://doi.org/10.1038/s41893-019-0305-3, 2019. Farinotti, D., Brinkerhoff, D. J., Clarke, G. K. C., Fürst, J. J.,
Frey, H., Gantayat, P., Gillet-Chaulet, F., Girard, C., Huss, M.,
Leclercq, P. W., Linsbauer, A., Machguth, H., Martin, C., Maus-
sion, F., Morlighem, M., Mosbeux, C., Pandit, A., Portmann,
A., Rabatel, A., Ramsankaran, R., Reerink, T. J., Sanchez,
O., Stentoft, P. A., Singh Kumari, S., van Pelt, W. J. J., An-
derson, B., Benham, T., Binder, D., Dowdeswell, J. A., Fis-
cher, A., Helfricht, K., Kutuzov, S., Lavrentiev, I., McNabb,
R., Gudmundsson, G. H., Li, H., and Andreassen, L. M.: How
accurate are estimates of glacier ice thickness? Results from
ITMIX, the Ice Thickness Models Intercomparison eXperi-
ment, The Cryosphere, 11, 949–970, https://doi.org/10.5194/tc-
11-949-2017, 2017. Bisset, R. R., Dehecq, A., Goldberg, D. N., Huss, M., Bing-
ham, R. G., and Gourmelen, N.: Reversed surface-mas-
balance gradients on Himalayan debris-covered glaciers in-
ferred from remote sensing, Remote Sensing, 12, 1563,
https://doi.org/10.3390/rs12101563, 2020. Bolch, T., Pieczonka, T., and Benn, D. I.: Multi-decadal mass
loss of glaciers in the Everest area (Nepal Himalaya) de-
rived from stereo imagery, The Cryosphere, 5, 349–358,
https://doi.org/10.5194/tc-5-349-2011, 2011. Brun, F., Berthier, E., Wagnon, P., Kääb, A., and Treichler, D.:
A spatially resolved estimate of High Mountain Asia glacier
mass balances from 2000 to 2016, Nat. Geosci., 10, 668–673,
https://doi.org/10.1038/ngeo2999, 2017. Farinotti, D., Huss, M., Fürst, J. J., Landmann, J., Machguth, H.,
Maussion, F., and Pandit, A.: A consensus estimate for the ice
thickness distribution of all glaciers on Earth, Nat. Geosci., 12,
168–173, https://doi.org/10.1038/s41561-019-0300-3, 2019a. References Anderson, L. S. and Anderson, R. S.: Modeling debris-covered
glaciers: response to steady debris deposition, The Cryosphere,
10, 1105–1124, https://doi.org/10.5194/tc-10-1105-2016, 2016. Dehecq, A., Gardner, A. S., Alexandrov, O., McMichael, S.,
Hugonnet, R., Shean, D., and Marty, M.: Automated processing
of declassified KH-9 Hexagon satellite images for global eleva-
tion change analysis since the 1970s, Front. Earth Sci., 8, 566–
802, https://doi.org/10.3389/feart.2020.566802, 2020. Anderson, L. S., Armstrong, W. H., Anderson, R. S., and
Buri, P.: Debris cover and the thinning of Kennicott Glacier,
Alaska: in situ measurements, automated ice cliff delineation
and distributed melt estimates, The Cryosphere, 15, 265–282,
https://doi.org/10.5194/tc-15-265-2021, 2021a. Deline, P. and Orombelli, G.: Glacier fluctuations in the western
Alps during the Neoglacial, as indicated by the Miage morainic
amphitheatre (Mont Blanc massif, Italy), Boreas, 34, 456–467,
https://doi.org/10.1111/j.1502-3885.2005.tb01444.x, 2005. Anderson, L. S., Armstrong, W. H., Anderson, R. S., Scher-
ler, D., and Petersen, E.: The causes of debris-covered glacier
thinning: Evidence for the Importance of ice dynamics from
Kennicott Glacier, Alaska, Front. Earth Sci., 9, 680995,
https://doi.org/10.3389/feart.2021.680995, 2021b. Dobhal, D., Mehta, M., and Srivastava, D.: Influence of debris
cover on terminus retreat and mass changes of Chorabari Glacier,
Garhwal region, central Himalaya, India, J. Glaciol., 59, 961–
971, https://doi.org/10.3189/2013JoG12J180, 2013. Anderson, R. S.: A model of ablation-dominated medial moraines
and the generation of debris-mantled glacier snouts, J. Glaciol.,
46,
459–469,
https://doi.org/10.3189/172756500781833025,
2000. Dunning,
S. A.,
Rosser,
N. J.,
McColl,
S. T.,
and
Reznichenko,
N. V.:
Rapid
sequestration
of
rock
avalanche deposits within glaciers, Nat. Commun., 6, 1–7,
https://doi.org/10.1038/ncomms8964, 2015. Benn, D., Bolch, T., Hands, K., Gulley, J., Luckman, A.,
Nicholson, L., Quincey, D., Thompson, S., Toumi, R., and
Wiseman, S.: Response of debris-covered glaciers in the
Mount Everest region to recent warming, and implications
for outburst flood hazards, Earth-Sci. Rev., 114, 156–174,
https://doi.org/10.1016/j.earscirev.2012.03.008, 2012. É Edwards, T. L., Nowicki, S., Marzeion, B., Hock, R., Goelzer,
H., Seroussi, H., Jourdain, N. C., Slater, D. A., Turner,
F. E., Smith, C. J., et al.: Projected land ice contributions
to twenty-first-century sea level rise, Nature, 593, 74–82,
https://doi.org/10.1038/s41586-021-03302-y, 2021. Emmer,
A.,
Vilímek,
V.,
Klimeš,
J.,
and
Cochachin,
A.:
Glacier retreat, lakes development and associated natural haz-
ards in Cordilera Blanca, Peru, in: Landslides in cold re-
gions in the context of climate change, Springer, 231–252,
https://doi.org/10.1007/978-3-319-00867-7_17, 2014. Berthier, É. L. Compagno et al.: Regional-scale modelling of supraglacial debris 1715 lution in Scandinavia and Iceland, J. Glaciol., 67, 727–743,
https://doi.org/10.1017/jog.2021.24, 2021. L. Compagno et al.: Regional-scale modelling of supraglacial debris We also suggest doing so not
only at the glacier-specific scale but also at the regional scale
– especially when addressing regions that feature a signifi-
cant share of debris-covered glaciers. We also promote fur-
ther investigations directed to the past evolution of debris-
covered area and thickness. This would be important for ac-
quiring further knowledge about the processes controlling
their evolution, as well as for better constraining some of the
necessary model parameters. We suggest that both remote- Review statement. This paper was edited by Andreas Vieli and re-
viewed by Leif S. Anderson and Ben Marzeion. https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022 L. Compagno et al.: Regional-scale modelling of supraglacial debris A.:
ITS-LIVE
Regional
Glacier
and
Ice
Sheet
Sur-
face
Velocities,
National
Snow
and
Ice
Data
Center,
https://doi.org/10.5067/6II6VW8LLWJ7, 2019. Juen, M., Mayer, C., Lambrecht, A., Han, H., and Liu, S.: Impact
of varying debris cover thickness on ablation: a case study for
Koxkar Glacier in the Tien Shan, The Cryosphere, 8, 377–386,
https://doi.org/10.5194/tc-8-377-2014, 2014. Gibson, M. J., Glasser, N. F., Quincey, D. J., Mayer, C.,
Rowan, A. V., and Irvine-Fynn, T. D.: Temporal variations
in supraglacial debris distribution on Baltoro Glacier, Karako-
ram between 2001 and 2012, Geomorphology, 295, 572–585,
https://doi.org/10.1016/j.geomorph.2017.08.012, 2017. Kayastha, R. B., Takeuchi, Y., Nakawo, M., and Ageta, Y.: Practi-
cal prediction of ice melting beneath various thickness of debris
cover on Khumbu Glacier, Nepal, using a positive degree-day
factor, IAHS-AISH P., 264, 71–81, 2000. Groos, A. R., Mayer, C., Smiraglia, C., Diolaiuti, G., and
Lambrecht, A.: A first attempt to model region-wide glacier
surface
mass
balances
in
the
Karakoram:
findings
and
future challenges, Geogr. Fis. Din. Quat., 40, 137–159,
https://doi.org/10.4461/GFDQ.2017.40.10, 2018. Khan, M. I.: Ablation on Barpu Glacier, Karakoram Himalaya, Pak-
istan a study of melt processes on a faceted, debris-covered ice
surface, MSc thesis, Wilfrid Laurier University, https://scholars. wlu.ca/etd/311 (last access: 15 February 2021), 1989. Hagg, W., Mayer, C., Lambrecht, A., and Helm, A.: Sub-
debris melt rates on southern Inylchek Glacier, central Tian
Shan, Geografiska Annaler: Series A, Phys. Geogr., 90, 55–63,
https://doi.org/10.1111/j.1468-0459.2008.00333.x, 2008. Kienholz, C., Hock, R., Truffer, M., Bieniek, P., and Lader, R.: Mass
Balance Evolution of Black Rapids Glacier, Alaska, 1980–2100,
and Its Implications for Surge Recurrence, Front. Earth Sci., 5,
00056, https://doi.org/10.3389/feart.2017.00056, 2017. Herreid,
S. and
Pellicciotti,
F.:
The
state
of
rock
debris
covering
Earth’s
glaciers,
Nat. Geosci.,
13,
621–627,
https://doi.org/10.1038/s41561-020-0615-0, 2020. Kirkbride, M. P.: Ice-marginal geomorphology and Holocene ex-
pansion of debris-covered Tasman Glacier, New Zealand, IAHS-
AISH P., 264, 211–218, 2000. Herreid, S., Pellicciotti, F., Ayala, A., Chesnokova, A., Kienholz,
C., Shea, J., and Shrestha, A.: Satellite observations show no net
change in the percentage of supraglacial debris-covered area in
northern Pakistan from 1977 to 2014, J. Glaciol., 61, 524–536,
https://doi.org/10.3189/2015JoG14J227, 2015. Kneib, M., Miles, E., Jola, S., Buri, P., Herreid, S., Bhattacharya,
A., Watson, C., Bolch, T., Quincey, D., and Pellicciotti, F.:
Mapping ice cliffs on debris-covered glaciers using multispec-
tral satellite images, Remote Sens. Environ., 253, 112–201,
https://doi.org/10.1016/j.rse.2020.112201, 2021. L. Compagno et al.: Regional-scale modelling of supraglacial debris 1716 in the early twenty-first century, Nature, 592, 726–731,
https://doi.org/10.1038/s41586-021-03436-z, 2021. in the early twenty-first century, Nature, 592, 726–731,
https://doi.org/10.1038/s41586-021-03436-z, 2021. Farinotti, D., Brinkerhoff, D. J., Fürst, J. J., Gantayat, P., Gillet-
Chaulet, F., Huss, M., Leclercq, P. W., Maurer, H., Morlighem,
M., Pandit, A., et al.: Results from the Ice Thickness Models In-
tercomparison eXperiment Phase 2 (ITMIX2), Front. Earth Sci.,
8, 571923, https://doi.org/10.3389/feart.2020.571923, 2021. Huss, M.: Density assumptions for converting geodetic glacier
volume change to mass change, The Cryosphere, 7, 877–887,
https://doi.org/10.5194/tc-7-877-2013, 2013. Huss, M. and Hock, R.: A new model for global glacier
change
and
sea-level
rise,
Front. Earth
Sci.,
3,
00054,
https://doi.org/10.3389/feart.2015.00054, 2015. Ferguson, J. C. and Vieli, A.: Modelling steady states and the tran-
sient response of debris-covered glaciers, The Cryosphere, 15,
3377–3399, https://doi.org/10.5194/tc-15-3377-2021, 2021. Fyffe, C., Brock, B., Kirkbride, M., Mair, D., Arnold, N.,
Smiraglia, C., Diolaiuti, G., and Diotri, F.: Do debris-
covered glaciers demonstrate distinctive hydrological behaviour
compared
to
clean
glaciers?,
J. Hydrol.,
570,
584–597,
https://doi.org/10.1016/j.jhydrol.2018.12.069, 2019. Huss, M., Jouvet, G., Farinotti, D., and Bauder, A.: Fu-
ture high-mountain hydrology: a new parameterization of
glacier
retreat,
Hydrol. Earth
Syst. Sci.,
14,
815–829,
https://doi.org/10.5194/hess-14-815-2010, 2010. Immerzeel, W. W., Lutz, A., Andrade, M., Bahl, A., Biemans, H.,
Bolch, T., Hyde, S., Brumby, S., Davies, B., Elmore, A., Em-
mer, A., Feng, M., Fernández, A., Haritashya, U., Kargel, J. S.,
Koppes, M., Kraaijenbrink, P. D. A., Kulkarni, A. V., Mayewski,
P. A., Nepal, S., Pacheco, P., Painter, T. H., Pellicciotti, F., Ra-
jaram, H., Rupper, S., Sinisalo, A., Shrestha, A. B., Viviroli, D.,
Wada, Y., Xiao, C., Yao, T., and Baillie, J. E. M.: Importance and
vulnerability of the world’s water towers, Nature, 577, 364–369,
2020. Fyffe, C. L., Woodget, A. S., Kirkbride, M. P., Deline, P., Westoby,
M. J., and Brock, B. W.: Processes at the margins of supraglacial
debris cover: Quantifying dirty ice ablation and debris redistri-
bution, Earth Surf. Proc. Land., 45, 2272–2290, 2020. Gardelle, J., Berthier, E., Arnaud, Y., and Kääb, A.: Region-
wide
glacier
mass
balances
over
the
Pamir-Karakoram-
Himalaya during 1999–2011, The Cryosphere, 7, 1263–1286,
https://doi.org/10.5194/tc-7-1263-2013, 2013. Jouvet,
G.,
Huss,
M.,
Funk,
M.,
and
Blatter,
H.:
Mod-
elling
the
retreat
of
Grosser
Aletschgletscher,
Switzer-
land, in a changing climate, J. Glaciol., 57, 1033–1045,
https://doi.org/10.3189/002214311798843359, 2011. Gardner,
A. S.,
Fahnestock,
M. A.,
and
Scambos,
T. References Buri, P., Miles, E. S., Steiner, J. F., Ragettli, S., and Pellic-
ciotti, F.: Supraglacial ice cliffs can substantially increase the
mass loss of debris-covered glaciers, Geophys. Res. Lett., 48,
e2020GL092150, https://doi.org/10.1029/2020GL092150, 2021. Farinotti,
D.,
Round,
V.,
Huss,
M.,
Compagno,
L.,
and
Zekollari, H.: Large hydropower and water-storage poten-
tial in future glacier-free basins, Nature, 575, 341–344,
https://doi.org/10.1038/s41586-019-1740-z, 2019b. Chand, M. B. and Watanabe, T.: Development of supraglacial
ponds in the Everest region, Remote Sensing, 11, 1058,
https://doi.org/10.3390/rs11091058, 2019. Farinotti, D., Immerzeel, W. W., de Kok, R. J., Quincey,
D. J., and Dehecq, A.: Manifestations and mechanisms of
the Karakoram glacier Anomaly, Nat. Geosci., 13, 8–16,
https://doi.org/10.1038/s41561-019-0513-5, 2020. Compagno, L., Zekollari, H., Huss, M., and Farinotti, D.: Lim-
ited impact of climate forcing products on future glacier evo- The Cryosphere, 16, 1697–1718, 2022 https://doi.org/10.5194/tc-16-1697-2022 L. Compagno et al.: Regional-scale modelling of supraglacial debris L. Compagno et al.: Regional-scale modelling of supraglacial debris Ragettli, S., Pellicciotti, F., Immerzeel, W., Miles, E., Petersen, L.,
Heynen, M., Shea, J., Stumm, D., Joshi, S., and Shrestha, A.:
Unraveling the hydrology of a Himalayan catchment through in-
tegration of high resolution in situ data and remote sensing with
an advanced simulation model, Adv. Water Resour., 78, 94–111,
https://doi.org/10.1016/j.advwatres.2015.01.013, 2015. Maussion, F., Butenko, A., Champollion, N., Dusch, M., Eis, J.,
Fourteau, K., Gregor, P., Jarosch, A. H., Landmann, J., Oesterle,
F., Recinos, B., Rothenpieler, T., Vlug, A., Wild, C. T., and
Marzeion, B.: The Open Global Glacier Model (OGGM) v1.1,
Geosci. Model Dev., 12, 909–931, https://doi.org/10.5194/gmd-
12-909-2019, 2019. McCarthy, M., Miles, E., Kneib, M., Buri, P., Fugger, S., and Pel-
licciotti, F.: Supraglacial debris thickness and supply rate in
High Mountain Asia, Communications Earth and Environment,
https://doi.org/10.31223/X5WW5B, preprint, 2021. Ragettli,
S.,
Bolch,
T.,
and
Pellicciotti,
F.:
Heterogeneous
glacier thinning patterns over the last 40 years in Lang-
tang
Himal,
Nepal,
The
Cryosphere,
10,
2075–2097,
https://doi.org/10.5194/tc-10-2075-2016, 2016. Mihalcea, C., Mayer, C., Diolaiuti, G., Lambrecht, A., Smi-
raglia,
C.,
and
Tartari,
G.:
Ice
ablation
and
meteoro-
logical conditions on the debris-covered area of Baltoro
glacier, Karakoram, Pakistan, Ann. Glaciol., 43, 292–300,
https://doi.org/10.3189/172756406781812104, 2006. Reid, T. D. and Brock, B. W.: An energy-balance model for debris-
covered glaciers including heat conduction through the debris
layer, J. Glaciol., 56, 903–916, 2010. Reznichenko, N., Davies, T., Shulmeister, J., and McSaveney, M.:
Effects of debris on ice-surface melting rates: an experimental
study, J. Glaciol., 56, 384–394, 2010a. Miles,
E. S.,
Willis,
I.,
Buri,
P.,
Steiner,
J. F.,
Arnold,
N. S., and Pellicciotti, F.: Surface pond energy absorp-
tion across four Himalayan glaciers accounts for 1/8 of to-
tal catchment ice loss, Geophys. Res. Lett., 45, 10–464,
https://doi.org/10.1029/2018GL079678, 2018. Reznichenko,
N.,
Davies,
T.,
Shulmeister,
J.,
and
Mc-
Saveney,
M.:
Effects
of
debris
on
ice-surface
melting
rates:
an
experimental
study,
J. Glaciol.,
56,
384–394,
https://doi.org/10.3189/002214310792447725, 2010b. RGI Consortium: Randolph Glacier Inventory 6.0, National Snow
and Ice Data Center (NSIDC), https://doi.org/10.7265/N5-RGI-
60, 2017. Miles, K. E., Hubbard, B., Miles, E. S., Quincey, D. J., Rowan,
A. V., Kirkbride, M., and Hornsey, J.: Continuous borehole opti-
cal televiewing reveals variable englacial debris concentrations at
Khumbu Glacier, Nepal, Communications Earth & Environment,
2, 1–9, https://doi.org/10.1038/s43247-020-00070-x, 2021. Rounce, D. R., King, O., McCarthy, M., Shean, D. L. Compagno et al.: Regional-scale modelling of supraglacial debris 1717 supraglacial lakes on debris-covered glaciers in the Tien
Shan Mountains, Central Asia, Geomorphology, 286, 133–142,
https://doi.org/10.1016/j.geomorph.2017.03.002, 2017. debris-cover evolution form the single glacier to the regional
scale: An application to High Mountain Asia, ETH Zurich [data
set], https://doi.org/10.3929/ETHZ-B-000544323, 2022, debris-cover evolution form the single glacier to the regional
scale: An application to High Mountain Asia, ETH Zurich [data
set], https://doi.org/10.3929/ETHZ-B-000544323, 2022, Marzeion, B., Jarosch, A. H., and Hofer, M.: Past and future sea-
level change from the surface mass balance of glaciers, The
Cryosphere, 6, 1295–1322, https://doi.org/10.5194/tc-6-1295-
2012, 2012. Nicholson, L. and Benn, D. I.: Calculating ice melt beneath a de-
bris layer using meteorological data, J. Glaciol., 52, 463–470,
https://doi.org/10.3189/172756506781828584, 2006. Østrem, G.: Ice melting under a thin layer of moraine, and the exis-
tence of ice cores in moraine ridges, Geogr. Ann., 41, 228–230,
https://doi.org/10.1080/20014422.1959.11907953, 1959. Marzeion, B., Kaser, G., Maussion, F., and Champollion, N.:
Limited influence of climate change mitigation on short-
term glacier mass loss, Nat. Clim. Change, 8, 305–308,
https://doi.org/10.1038/s41558-018-0093-1, 2018. Otsu, N.: A threshold selection method from gray-level histograms,
IEEE T. Syst. Man Cyb., 9, 62–66, 1979. Marzeion, B., Hock, R., Anderson, B., Bliss, A., Champollion, N.,
Fujita, K., Huss, M., Immerzeel, W., Kraaijenbrink, P., Malles, J.-
H., Maussion, F., Radic, V., Rounce, D. R., Sakai, A., Shannon,
S., van de Wal, R. S. W., and Zekollari, H.: Results of the sec-
ond experiment of the Glacier Model Intercomparison Project,
PANGAEA, https://doi.org/10.1594/PANGAEA.914503, 2020. Owen, L. A., Derbyshire, E., and Scott, C. H.: Contempo-
rary sediment production and transfer in high-altitude glaciers,
Sediment. Geol., 155, 13–36, https://doi.org/10.1016/S0037-
0738(02)00156-2, 2003. Pellicciotti,
F.,
Stephan,
C.,
Miles,
E.,
Herreid,
S.,
Im-
merzeel,
W. W.,
and
Bolch,
T.:
Mass-balance
changes
of
the
debris-covered
glaciers
in
the
Langtang
Himal,
Nepal,
from
1974
to
1999,
J. Glaciol.,
61,
373–386,
https://doi.org/10.3189/2015JoG13J237, 2015. Mattson, L. and Gardner, J.: Energy exchanges and ablation rates
on the debris-covered Rakhiot Glacier, Pakistan, Zeitschrift für
Gletscherkunde und Glazialgeologie, 25, 17–32, 1989. Radi´c, V. and Hock, R.: Glaciers in the earth hydrological
cycle: Assessments of glacier mass and runoff changes on
global and regional scales, Surv. Geophys., 35, 813–837,
https://doi.org/10.1007/s10712-013-9262-y, 2014. Maurer, J. and Rupper, S.: Tapping into the Hexagon spy
imagery database: A new automated pipeline for geomor-
phic change detection, ISPRS J. Photogramm., 108, 113–127,
https://doi.org/10.1016/j.isprsjprs.2015.06.008, 2015. L. Compagno et al.: Regional-scale modelling of supraglacial debris Hersbach, H., Bell, W., Berrisford, P., Horànyi, A., Muñoz-
Sabate, J., Nicolas, J., Radu, R., Schepers, D., Simmons,
A., Soci, C., and Dee, D.: Global reanalysis: goodbye
ERA-Interim, hello ERA5, ECMWF, 3, e2011834–e2011834,
https://doi.org/10.21957/vf291hehd7, 2019. Kraaijenbrink, P. D. A., Bierkens, M. F. P., Lutz, A. F., and
Immerzeel, W. W.: Impact of a global temperature rise of
1.5 degrees Celsius on Asia’s glaciers, Nature, 549, 257–260,
https://doi.org/10.1038/nature23878, 2017. Hock, R.: Temperature index melt modelling in mountain ar-
eas, J. Hydrol., 282, 104–115, https://doi.org/10.1016/S0022-
1694(03)00257-9, 2003. Lee, K., Fyson, C., and Schleussner, C.-F.: Fair distributions of
carbon dioxide removal obligations and implications for effec-
tive national net-zero targets, Environ. Res. Lett., 16, 094001,
https://doi.org/10.1088/1748-9326/ac1970, 2021. Hugonnet, R., McNabb, R., Berthier, E., Menounos, B., Nuth,
C., Girod, L., Farinotti, D., Huss, M., Dussaillant, I., Brun,
F., and Kääb, A.: Accelerated global glacier mass loss Loris, C., Huss, M., Miles, E. S., McCarthy, M. J., Zekollari, H., De-
hecq, A., Pellicciotti, F., and Daniel, F.: Modelling supraglacial The Cryosphere, 16, 1697–1718, 2022 https://doi.org/10.5194/tc-16-1697-2022 L. Compagno et al.: Regional-scale modelling of supraglacial debris Wei, Y., Tandong, Y., Baiqing, X., and Hang, Z.: Influence of
supraglacial debris on summer ablation and mass balance in the
24K Glacier, southeast Tibetan Plateau, Geogr. Ann. A, 92, 353–
360, https://doi.org/10.1111/j.1468-0459.2010.00400.x, 2010. Shannon, S., Smith, R., Wiltshire, A., Payne, T., Huss, M., Betts,
R., Caesar, J., Koutroulis, A., Jones, D., and Harrison, S.: Global
glacier volume projections under high-end climate change sce-
narios, The Cryosphere, 13, 325–350, https://doi.org/10.5194/tc-
13-325-2019, 2019. WGMS: Fluctuations of glaciers database, World Glacier Monitor-
ing Service, https://doi.org/10.5904/wgms-fog-2020-08, 2020. Wirbel, A., Jarosch, A. H., and Nicholson, L.: Modelling debris
transport within glaciers by advection in a full-Stokes ice flow
model, The Cryosphere, 12, 189–204, https://doi.org/10.5194/tc-
12-189-2018, 2018. Sharma, P., Patel, L. K., Ravindra, R., Singh, A., Mahalinganathan,
K., and Thamban, M.: Role of debris cover to control specific
ablation of adjoining Batal and Sutri Dhaka glaciers in Chan-
dra Basin (Himachal Pradesh) during peak ablation season, J. Earth Syst. Sci., 125, 459–473, https://doi.org/10.1007/s12040-
016-0681-2, 2016. Wyser,
K.,
Kjellström,
E.,
Koenigk,
T.,
Martins,
H.,
and
Döscher, R.: Warmer climate projections in EC-Earth3-Veg:
The role of changes in the greenhouse gas concentrations
from CMIP5 to CMIP6, Environ. Res. Lett., 15, 054020,
https://doi.org/10.1088/1748-9326/ab81c2, 2020. Shean, D. E., Bhushan, S., Montesano, P., Rounce, D. R., Arendt,
A., and Osmanoglu, B.: Regional assessment of High Moun-
tain asia glacier mass balance, Front. Earth Sci., 7, 363,
https://doi.org/10.3389/feart.2019.00363, 2020. Zekollari, H., Huss, M., and Farinotti, D.: Modelling the future
evolution of glaciers in the European Alps under the EURO-
CORDEX RCM ensemble, The Cryosphere, 13, 1125–1146,
https://doi.org/10.5194/tc-13-1125-2019, 2019. Shugar, D. H. and Clague, J. J.: The sedimentology and
geomorphology
of
rock
avalanche
deposits
on
glaciers,
Sedimentology, 58, 1762–1783, https://doi.org/10.1111/j.1365-
3091.2011.01238.x, 2011. Zekollari, H., Huss, M., and Farinotti, D.: On the imbal-
ance and response time of glaciers, Geophys. Res. Let., 47,
e2019GL085578, https://doi.org/10.1029/2019GL085578, 2020. Shukla, A. and Qadir, J.: Differential response of glaciers
with varying debris cover extent: evidence from changing
glacier parameters, Int. J. Remote Sens., 37, 2453–2479,
https://doi.org/10.1080/01431161.2016.1176272, 2016. Zemp, M., Huss, M., Thibert, E., Eckert, N., McNabb, R., Huber,
J., Barandun, M., Machguth, H., Nussbaumer, S. U., Gartner-
Roer, I., Thomson, L., Paul, F., Maussion, F., Kutuzov, S., and
Cogley, J. G.: Global glacier mass changes and their contribu-
tions to sea-level rise from 1961 to 2016, Nature, 568, 382–386,
https://doi.org/10.1038/s41586-019-1071-0, 2019. Steiner, J. F., Litt, M., Stigter, E. E., Shea, J., Bierkens,
M. F., and Immerzeel, W. L. Compagno et al.: Regional-scale modelling of supraglacial debris 1718 Thickness Estimates Reveal Debris Significantly Impacts Glacier
Mass Balance, Geophys. Res. Lett., 48, e2020GL091311,
https://doi.org/10.1029/2020GL091311, 2021. Tangborn, W. and Rana, B.: Mass balance and runoff of the par-
tially debris-covered Langtang Glacier, Nepal, IAHS-AISH P.,
264, 99–108, 2000. Tielidze, L. G., Bolch, T., Wheate, R. D., Kutuzov, S. S., Lavrentiev,
I. I., and Zemp, M.: Supra-glacial debris cover changes in the
Greater Caucasus from 1986 to 2014, The Cryosphere, 14, 585–
598, https://doi.org/10.5194/tc-14-585-2020, 2020. Rowan, A. V., Egholm, D. L., Quincey, D. J., and Glasser, N. F.:
Modelling the feedbacks between mass balance, ice flow and de-
bris transport to predict the response to climate change of debris-
covered glaciers in the Himalaya, Earth Planet. Sc. Lett., 430,
427–438, https://doi.org/10.1016/j.epsl.2015.09.004, 2015. Van de Wal, R. S. W. and Wild, M.: Modelling the response of
glaciers to climate change by applying volume-area scaling in
combination with a high resolution GCM, Clim. Dynam., 18,
359–366, https://doi.org/10.1007/s003820100184, 2001. Sakai, A. and Fujita, K.: Contrasting glacier responses to recent
climate change in high-mountain Asia, Sci. Rep.-UK, 7, 13717,
https://doi.org/10.1038/s41598-017-14256-5, 2017. Van Woerkom, T., Steiner, J. F., Kraaijenbrink, P. D. A., Miles, E. S.,
and Immerzeel, W. W.: Sediment supply from lateral moraines to
a debris-covered glacier in the Himalaya, Earth Surf. Dynam., 7,
411–427, https://doi.org/10.5194/esurf-7-411-2019, 2019. Sakai, A., Nakawo, M., and Fujita, K.: Melt rate of ice cliffs on the
Lirung Glacier, Nepal Himalayas, 1996, Bull. Glacier Res, 16,
57–66, 1998. Sakai, A., Takeuchi, N., Fujita, K., and Nakawo, M.: Role of
supraglacial ponds in the ablation process of a debris-covered
glacier in the Nepal Himalayas, IAHS-AISH P., 265, 119–132,
2000. Verhaegen, Y., Huybrechts, P., Rybak, O., and Popovnin, V. V.:
Modelling the evolution of Djankuat Glacier, North Cauca-
sus, from 1752 until 2100 CE, The Cryosphere, 14, 4039–4061,
https://doi.org/10.5194/tc-14-4039-2020, 2020. Scherler, D. and Egholm, D.: Production and transport of
supraglacial debris: Insights from cosmogenic 10Be and
numerical
modeling,
Chhota
Shigri
Glacier,
Indian
Hi-
malaya,
J. Geophys. Res.-Earth,
125,
e2020JF005586,
https://doi.org/10.1029/2020JF005586, 2020. Watson, C. S., Quincey, D. J., Carrivick, J. L., and Smith,
M. W.:
Ice
cliff
dynamics
in
the
Everest
region
of
the
Central
Himalaya,
Geomorphology,
278,
238–251,
https://doi.org/10.1016/j.geomorph.2016.11.017, 2017. Scherler, D., Wulf, H., and Gorelick, N.: Global assessment
of supraglacial debris-cover extents, Geophys. Res. Lett., 45,
11798–11805, https://doi.org/10.1029/2018GL080158, 2018. L. Compagno et al.: Regional-scale modelling of supraglacial debris E., and Salerno,
F.: Quantifying debris thickness of debris-covered glaciers in
the Everest Region of Nepal through inversion of a subde-
bris melt model, J. Geophys. Res.-Earth, 123, 1094–1115,
https://doi.org/10.1029/2017JF004395, 2018. Mölg, N., Bolch, T., Walter, A., and Vieli, A.: Unravelling the
evolution of Zmuttgletscher and its debris cover since the
end of the Little Ice Age, The Cryosphere, 13, 1889–1909,
https://doi.org/10.5194/tc-13-1889-2019, 2019. Rounce, D. R., Hock, R., and Shean, D.: Glacier mass change in
High Mountain Asia through 2100 using the open-source Python
Glacier Evolution Model (PyGEM), Front. Earth Sci., 7, 00331,
https://doi.org/10.3389/feart.2019.00331, 2020. Mölg, N., Ferguson, J., Bolch, T., and Vieli, A.: On the influ-
ence of debris cover on glacier morphology: How high-relief
structures evolve from smooth surfaces, Geomorphology, 357,
107092, https://doi.org/10.1016/j.geomorph.2020.107092, 2020. Rounce, D. R., Hock, R., McNabb, R. W., Millan, R., Som-
mer, C., Braun, M. H., Malz, P., Maussion, F., Mouginot, J.,
Seehaus, T. C., and Shean, D. E.: Distributed Global Debris Narama, C., Daiyrov, M., Tadono, T., Yamamoto, M., Kääb,
A.,
Morita,
R.,
and
Ukita,
J.:
Seasonal
drainage
of https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022 L. Compagno et al.: Regional-scale modelling of supraglacial debris W.: The importance of tur-
bulent fluxes in the surface energy balance of a debris-
covered glacier in the Himalayas, Front. Earth Sci., 6, 144,
https://doi.org/10.3389/feart.2018.00144, 2018. Zheng, G., Allen, S. K., Bao, A., Ballesteros-Cánovas, J. A., Huss,
M., Zhang, G., Li, J., Yuan, Y., Jiang, L., Yu, T., Chen, W., and
Stoffel, M.: Increasing risk of glacial lake outburst floods from
future Third Pole deglaciation, Nat. Clim. Change, 11, 411–417,
https://doi.org/10.1038/s41558-021-01028-3, 2021. Stokes, C., Popovnin, V., Aleynikov, A., Gurney, S., and
Shahgedanova, M.: Recent glacier retreat in the Caucasus Moun-
tains, Russia, and associated increase in supraglacial debris cover
and supra-/proglacial lake development, Ann. Glaciol., 46, 195–
203, https://doi.org/10.3189/172756407782871468, 2007. https://doi.org/10.5194/tc-16-1697-2022 The Cryosphere, 16, 1697–1718, 2022
|
https://openalex.org/W2606746047
|
https://revistarebram.com/index.php/revistauniara/article/download/146/117
|
Portuguese
| null |
Amelogênese Imperfeita, Hipoplasia de Esmalte e Fluorose Dental – Revisão da Literatura
|
Revista Brasileira Multidisciplinar
| 2,010
|
cc-by
| 7,824
|
PALAVRAS-CHAVE: Amelogênese imperfeita; Hipoplasia de esmalte dentário; Fluorose dentária; Esmalte dentário. PALAVRAS-CHAVE: Amelogênese imperfeita; Hipoplasia de esmalte dentário; Fluorose dentária; Esmalte dentário. AMELOGÊNESE IMPERFEITA, HIPOPLASIA DE ESMALTE E
FLUOROSE DENTAL – REVISÃO DA LITERATURA BEVILACQUA, Flávia Magnani. Mestre e doutora em Dentística. Professora de Dentística da
Universidade Paulista – Unip/Campinas e do Centro Universitário de Araraquara – Uniara. SACRAMENTO, Tamires. Aluna do curso de Odontologia da Universidade Paulista – Unip/Campinas. FELÍCIO, Cristina Magnani. Mestre e doutora em Dentística. Professora de Dentística do Centro
Universitário de Araraquara – Uniara. Endereço para correspondência: Av. Mauá, 780, apto 32 CEP 14801-190
Araraquara-SP. E-mail: crismagnani@hotmail.com. RESUMO Os distúrbios de desenvolvimento no esmalte apresentam-se como anomalias de estrutura, podendo afetar
ambas as dentições. Dentre eles estão a amelogênese imperfeita, a hipoplasia de esmalte e a fluorose dental. A
amelogênese imperfeita é uma alteração de caráter hereditário e a hipoplasia de esmalte, um defeito quantitativo
do esmalte que ocorre em consequência de problemas sistêmicos, locais e hereditários, ou até da combinação
dos mesmos. A fluorose dental é uma hipoplasia gerada pela ingestão crônica de flúor durante o período da
odontogênese. Todas essas alterações apresentam características clínicas semelhantes, sendo necessário muito
cuidado na sua avaliação. É de extrema importância o conhecimento dessas anomalias para que o diagnóstico
diferencial seja instituído e, consequentemente, o plano de tratamento seja estabelecido de forma apropriada
para cada situação. Sendo assim, o objetivo do presente trabalho foi realizar uma revisão da literatura a respeito
dessas três anomalias do esmalte dental. Por meio desta revisão, concluiu-se que, para estabelecer o diagnóstico
diferencial entre essas alterações, assim como um correto plano de tratamento, é necessário o conhecimento
das anomalias pelo profissional associado à execução de exame clínico, composto de anamnese e exame físico
e, em alguns casos, de exame radiográfico. artigo de revisão REVISÃO DA LITERATURA Nas anomalias de formação do esmalte ocorre
alteração da cor do dente, o que compromete a estética,
algo muito importante para a população atualmente. Em
geral, as alterações de cor do dente podem ser de origem
extrínseca, relacionadas ao consumo de medicamentos,
bebidas ou alimentos que possuem corantes naturais ou
artificiais em sua composição, ou intrínseca, causadas
por fatores sistêmicos, como a genética, deficiências
congênitas, falhas de metabolismo pré-natal, deficiência
nutricional, entre outros fatores. A severidade das
manchas dependerá da fase de desenvolvimento e do
tempo em que o fator etiológico atuou durante a
formação do esmalte dentário, e o tratamento dependerá
do diagnóstico correto das anomalias. Os defeitos
decorrentes da amelogenêse imperfeita, hipoplasia e
fluorose são alterações na estrutura do esmalte, as quais
causam as manchas intrínsecas, apresentando tratamento
mais complexo, apresentando-se algumas vezes como
uma mancha branca leitosa, de aspecto liso e brilhante,
ou como manchas de coloração escura. Para um bom
resultado, o diagnóstico correto das manchas é tão
importante quanto o tratamento (MACHADO, 2001). Um dos motivos da sensibilidade é o fato dos
ameloblastos serem células epiteliais, com grande
síntese proteica. Portanto, alterações nutricionais,
distúrbios de perfusão vascular, bem como distúrbios
no metabolismo de cálcio levam a alterações do
esmalte. Alterações mais severas podem também estar
relacionadas com menor tempo de gestação, asfixia
neonatal, baixo peso ao nascimento e desnutrição
(GUERLACH, 2000). Os distúrbios de desenvolvimento no esmalte
apresentam-se como anomalias de estrutura, podendo
afetar ambas as dentições (PITHAN, 2002). Dentre
eles estão a hipoplasia de esmalte, a amelogênese
imperfeita e a fluorose dental (PITHAN, 2002). Todas essas alterações apresentam características
clínicas semelhantes, sendo necessário um exame
clínico minucioso, além da anamnese cuidadosa e
exame radiográfico em alguns casos. É de extrema
importância o conhecimento dessas anomalias pelo
cirurgião-dentista para que o diagnóstico diferencial
seja instituído e, consequentemente, o plano de
tratamento seja estabelecido de forma apropriada para
cada situação. Sendo assim, o objetivo do presente
trabalho é realizar uma revisão da literatura a respeito
das anomalias do esmalte dental: amelogênese
imperfeita, hipoplasia e fluorose. Essas alterações, além de comprometerem a
estética, aumentam a sensibilidade dentária, favorecem
má oclusão e podem levar à perda da camada
protetora do esmalte , predispondo o dente à cárie
(HOFFMANN, 2007; OLIVEIRA, 2002). ABSTRACT The developmental disorders of enamel are abnormalities of structure which can affect both dentitions. These
abnormalities include amelogenesis imperfecta, enamel hypoplasia and dental fluorosis. The amelogenesis imperfecta
is a hereditary change and enamel hypoplasia is a quantitative defect of enamel that occurs as a result of systemic
problems, local and also inherited factors, or even the combination of them. Dental fluorosis is a hypoplasia
caused by the chronic ingestion of fluoride during odontogenesis. All these anomalies have similar clinical
characteristics, and it is necessary to be careful in their assessment. It is extremely important to know these
abnormalities to establish a differential diagnosis and, consequently, a treatment plan, which can be set for each
situation. Therefore, the purpose of this study was to review the literature regarding these three anomalies:
amelogenesis imperfecta, enamel hypoplasia and dental fluorosis. It was concluded that to establish the differential
diagnosis of these abnormalities as well as a proper treatment plan, it is indispensable the professional knowledge
associated with the clinical examination. The examination has to consist of medical history and physical examination,
and in some cases, x-ray examination. KEYWORDS: Amelogenesis imperfecta; Dental enamel hypoplasia; Dental fluorosis; Dental enamel. REVISTA UNIARA, v.13, n.2, dezembro 2010 136 Amelogênese imperfeita, hipoplasia de esmalte... INTRODUÇÃO amelogênese imperfeita, hipoplasia de esmalte e
fluorose dental. O esmalte dental é o tecido mais mineralizado do
corpo humano e sua formação pode ser dividida em
três estágios: formação da matriz celular, calcificação
e maturação (RIBAS, 2004). Qualquer alteração em
algum desses estágios pode gerar algum tipo de
anomalia de desenvolvimento dental (BRAGA, 2005),
ou seja, uma variação ou desvio de uma característica
relativa à normalidade, pois os ameloblastos, células
responsáveis pela formação do esmalte, constituem um
dos grupos de células mais sensíveis do corpo no que
diz respeito à função metabólica (PITHAN, 2002). REVISÃO DA LITERATURA Além
disso, todos os tipos de alterações no esmalte devem
ser cuidadosamente investigadas, considerando que o
mesmo, além de ser um importante marcador biológico
da história passada da doença cárie, também pode
fornecer subsídios e indicativos para investigar outras
doenças e fatores de exposição ambientais
(HOFFMANN, 2007). O frequente aparecimento de defeitos nos tecidos
dentais, muitas vezes comprometendo a integridade do
dente em diferentes graus de severidade, tem levado
estudiosos a pesquisarem suas possíveis causas. Vários
estudos demonstraram que esses defeitos podem estar OBJETIVOS O objetivo deste trabalho foi realizar uma revisando
de literatura a respeito de algumas das principais
anomalias que acometem o esmalte dental: REVISTA UNIARA, v.13, n.2, dezembro 2010 137 BEVILACQUA et al. relacionados a distúrbios sistêmicos sofridos pela
gestante, a problemas que afetaram a própria criança
no período perinatal ou nos seus primeiros meses de
vida. As crianças prematuras apresentam mais defeitos
em seus dentes (em ambas as dentições); a face
vestibular é a mais afetada; a idade gestacional interfere
na prevalência de defeitos de esmalte de dentes
decíduos; e a frequência de opacidades é maior que a
de hipoplasias (MACEDO 2003). hereditários do esmalte, não havendo associação com
quaisquer outros defeitos generalizados. Esta patologia
pode ser de origem autossômica dominante,
autossômica recessiva ou ligada ao cromossomo X. O desenvolvimento do esmalte normal ocorre em três
etapas: etapa formativa, na qual há deposição da matriz
orgânica; etapa de mineralização, em que a matriz é
parcialmente mineralizada; etapa de maturação,
durante a qual os cristais minerais aumentam e se
completam. Sendo assim, são relatados três tipos
básicos de amelogênese imperfeita, dependendo da
etapa de formação do esmalte dental em que ocorre:
tipo hipoplásico, em que há formação deficiente da
matriz; tipo hipomineralizado, em que há mineralização
deficiente da matriz formada; tipo hipomaturação, em
que os cristais de esmalte permanecem imaturos. O
paciente portador de amelogênese imperfeita pode
apresentar as coroas dentárias com ou sem alteração
de cor. Já o esmalte pode estar totalmente ausente, ter
variação quanto a sua textura, consistência ou até
mesmo ser relativamente duro (SHAFER, 1987). O defeito de desenvolvimento do esmalte pode ser
definido como distúrbio nas matrizes dos tecidos duros
e em sua mineralização ocorrido durante a
odontogênese, devido a alterações sistêmicas, como
processos infecciosos, uso de tetraciclina, distúrbios
metabólicos, alta ingestão de flúor e deficiências
nutricionais. Foi realizado um estudo epidemiológico
dos defeitos de desenvolvimento do esmalte e as
possíveis associações dessas alterações com o estado
nutricional de escolares de 7 a 10 anos de idade,
considerando o número de dentes afetados e a etiologia
em uma amostra de conveniência composta por 449
crianças da cidade de Botucatu, SP. Não houve
associação entre o número de dentes afetados pelos
defeitos de desenvolvimento do esmalte e o estado
nutricional; entretanto, verificou-se associação entre a
etiologia sistêmica e a ocorrência dos mesmos
(HANSER-DUCATTI, 2004). OBJETIVOS A amelogênese imperfeita afeta ambas as dentições,
mas é uma alteração hereditária que não tem associação
com quaisquer outros defeitos generalizados. Nela os
dentes se apresentam de cor acastanhada-escura,
devido à transparência da dentina adjacente, e ocorre
na população em geral na proporção de 1 a cada 15
mil indivíduos, em média. O processo de formação do esmalte dentário pode
ser alterado em razão do caráter genético e do caráter
ambiental. O esmalte malformado poderá apresentar
áreas hipoplásicas, um defeito quantitativo do esmalte,
e/ou hipocalcificadas, um defeito quantitativo do esmalte. Dependendo de sua extensão, poderá comprometer o
desempenho da função dental. O conhecimento do
padrão de comprometimento por ela apresentado fará
com que o tratamento seja mais facilmente planejado e
mais efetivo (RUSCHEL, 2006). Radiograficamente, os dentes com amelogenêse
imperfeita apresentam falta da imagem que delimita o
esmalte. Esses dentes são mais suscetíveis à atrição, o
que ocasiona perda da dimensão vertical, assim como
podem ser mais sensíveis à variação de temperatura,
podendo acarretar impulso lingual, ocasionando
mordida aberta anterior, na maioria dos casos. Existem
alguns estudos que sugerem que esses tipos de
pacientes tem menor chance de desenvolver cáries
dentais, pela ausência de contato proximal. Por outro
lado, há maior chance de desenvolver esse tipo de
doença pelo aumento na retenção de placa, devido à
rugosidade da superfície do esmalte nessa alteração. O tratamento dependerá do tipo de amelogênese
imperfeita, da gravidade e da necessidade de melhorar Um dos defeitos que podem afetar o esmalte
dentário é a amelogênese imperfeita, sendo um
distúrbio exclusivamente ectodérmico, pois os
componentes mesodérmicos do dente (que constituirão
dentina e polpa) se encontram basicamente normais. A amelogênese imperfeita constitui um grupo de defeitos REVISTA UNIARA, v.13, n.2, dezembro 2010 138 Amelogênese imperfeita, hipoplasia de esmalte... as condições estéticas, funcionais e psicológicas do
paciente. Geralmente consiste na confecção de coroas
totais metálicas, metalocerâmicas ou de resina
composta indireta, tendo ainda como opção o uso de
coroas de dentes extraídos e fixadas com resina
composta (PITHAN, 2002). sucesso do tratamento dessa anomalia. Observaram
também que o diagnóstico de defeitos no esmalte dental
deve basear-se em dados clínicos, radiográficos e,
quando possível, em dados laboratoriais, sendo que
somente o cuidadoso planejamento, associado a um
adequado acompanhamento, permitirá que o paciente
alcance uma dentição com características estético-
funcionais satisfatórias. A amelogênese imperfeita pode acometer a dentição
decídua e permanente, como resultado da deposição
inadequada dos cristais de hidroxiapatita. OBJETIVOS Essa
alteração pode causar retenção excessiva de matéria
orgânica, em que o processo da maturação será
interrompido. Existem três tipos de amelogênese
imperfeita: a hipoplásica (tipo 1) – espessura do esmalte
está reduzida em pontos ou áreas da superfície;
hipomaturada (tipo 2) – esmalte tem espessura normal,
porém é mais macio; hipocalcificada (tipo 3) – esmalte
tem espessura normal, mas é muito macio, opaco, com
coloração que varia do branco até o castanho-escuro. Dependendo do tipo da amelogênese imperfeita, os
dentes podem ter extrema sensibilidade a estímulos
térmicos e químicos (AUGUSTO, 2005). A hipoplasia de esmalte é uma alteração que ocorre
como consequência de problemas sistêmicos, locais
e hereditários, que afetam ambas as dentições, e é
caracterizada por uma formação incompleta ou
deficiente da matriz orgânica do esmalte. A hipoplasia
afeta a espessura do esmalte (OLIVEIRA,
ROSENBLAT, 2002). O aparecimento de hipoplasia de esmalte e
dilaceração coronária resulta de traumatismos de
dentes decíduos. A hipoplasia interna do esmalte pode
apresentar-se como uma descoloração branca ou
amarelo amarronzada, que é uma manifestação da
calcificação insuficiente durante o estágio de maturação
do esmalte. A etiologia inclui fatores hereditários,
doenças sistêmicas, administração de tetraciclina e
traumas mecânicos, sendo que este último pode resultar
em depressões e ranhuras. Radiograficamente, as áreas
hipoplásicas aparecem radiolúcidas (WATANABE,
2003). Os autores apresentam um caso clínico de uma
paciente que apresenta hipoplasia de esmalte e
dilaceração coronária, em que o tratamento executado
com restauração em resina composta fotopolimerizável,
associada a pino de fibra intrarradicular, foi favorável. Para os autores, é importante saber diagnosticar e
classificar corretamente esta anomalia para realizar um
tratamento adequado. O tratamento de casos de
amelogênese imperfeita pode ser parte de um plano
complexo de restauração e depende da severidade da
condição, podendo existir uma inter-relação de
disciplinas, combinando terapia ortodôntica, cirurgia
ortognática, cirurgia periodontal e prótese dental. Relatam um caso clínico de uma criança com
amelogênese imperfeita do tipo hipoplásica nas arcadas
superior e inferior, em que o tratamento realizado foi
restauração direta com resina fotopolimerizável . Os defeitos de esmalte dental são comuns, com
sua incidência variando entre as populações. A
hipoplasia de esmalte é o defeito mais frequente,
podendo acometer ambas as dentições. Turner, em
1912, foi o primeiro a descrever a hipoplasia
associada a fatores locais, o que levou este tipo de
alteração a ficar conhecida como Dente de Turner
(BRAGA et al., 2005). REVISTA UNIARA, v.13, n.2, dezembro 2010 OBJETIVOS Em sua pesquisa os autores
descrevem um caso clínico de um paciente pediátrico
que relatava dor provocada no elemento 45,
estruturalmente displásico. Inicialmente foi feita a
ulectomia a laser e, depois, a restauração direta com Um caso clínico de uma criança com amelogênese
imperfeita do tipo hipoplásica foi descrito na literatura
(BRUSCO, 2008), em que o tratamento proposto foi
instrução de higiene oral, restaurações dos elementos
dentais afetados com resina composta fotopolimerizável
e confecção de placa inferior em resina acrílica, para
aumentar a dimensão vertical. Os autores observaram
que a amelogênese imperfeita pode apresentar quadros
complexos, sendo o diagnóstico correto e precoce e o
tratamento restaurador e preventivo essenciais para o REVISTA UNIARA, v.13, n.2, dezembro 2010 139 BEVILACQUA et al. resina composta fotopolimerizável. Concluíram que,
em muitos casos, é necessário tratamento restaurador
por razões estéticas e funcionais, sendo que os
materiais ionoméricos são adequados para o
tratamento de crianças e adolescentes, reforçando a
importância de um bom exame clínico, devido à
diversidade dos fatores etiológicos. pela perda da estrutura dentária, ou mesmo pela
formação imperfeita do esmalte. No entanto, ainda são
necessários outros estudos para relacionar esses defeitos
de esmalte à ocorrência de cárie dentária, associando
também outros fatores, como nível socioeconômico,
genética, hábitos de higiene oral e dieta. O tratamento da hipoplasia de esmalte inclui desde
técnicas de microabrasão dental até restaurações
diretas ou indiretas. Dentre os fatores ambientais que
podem interferir na formação do esmalte estão as
deficiências nutricionais, principalmente das vitaminas
A, C e D, as doenças exantemáticas, como sarampo,
varicela e escarlatina, a sífilis congênita, a hipocalcemia,
traumas por ocasião do nascimento, a prematuridade
e a eritroblastose fetal, as infecções ou trauma local, a
ingestão de substâncias químicas, como o flúor, a
radiação e as causas idiopáticas. Essa alteração se
caracteriza como superfícies dentárias de fossetas,
ranhuras ou áreas maiores de esmalte perdido, podendo
ser regulares ou irregulares (BENDO, 2007). Os
autores relatam um caso de uma criança que
apresentava hipoplasia de esmalte nos dentes incisivos,
queixando-se de estética deficiente, onde o tratamento
realizado foi constituído de restaurações diretas com
resina composta. Concluíram que essa alternativa de
tratamento para lesões hipoplásicas no esmalte dentário
se mostrou eficiente no restabelecimento da estética e
harmonia facial. g
Qualquer distúrbio ocorrido no período de
desenvolvimento dental pode gerar defeitos de
esmalte, como as hipoplasias e as opacidades. REVISTA UNIARA, v.13, n.2, dezembro 2010 OBJETIVOS A
hipoplasia de esmalte é uma deficiência na quantidade
do esmalte que pode ocorrer na forma de fóssulas,
fissuras ou perda de áreas grandes de esmalte. Na
hipoplasia ocorre uma formação incompleta ou
defeituosa da matriz orgânica do esmalte dentário. Isso ocorre em razão de algum estímulo, genético ou
ambiental, sobre os ameloblastos do germe dentário
durante seu desenvolvimento, sendo que a localização
e extensão do defeito do esmalte vai depender da
intensidade e duração do estímulo, e o tratamento
varia de acordo com a extensão e o tipo de lesão. A
opacidade é uma deficiência na mineralização do
esmalte, podendo ser demarcada ou difusa. Quando
é demarcada, apresenta espessura normal do esmalte
com superfície intacta, mas com translucidez de grau
variável, limites nítidos e claros, com coloração que
varia entre o bege, branco, amarelo ou marrom. Quando se apresenta difusa, também é uma alteração
na translucidez, de grau e cor variáveis, podendo ser
linear ou em placas, ou ter distribuição confluente,
mas que não apresenta limites claros. Tanto a hipoplasia do esmalte quanto a fluorose
dental resultam em lesões no esmalte dentário
caracterizadas por manchas brancas locais ou
generalizadas, as quais prejudicam a estética por
destoar do aspecto natural do esmalte dental. Essas
alterações podem ocorrer devido a fatores ambientais,
idiopáticos ou de natureza hereditária. É essencial o
diagnóstico diferencial dessas alterações para o
estabelecimento do correto diagnóstico e tratamento,
que pode variar desde a fluorterapia até a confecção
de facetas de resina composta, dependendo do
comprometimento estético e/ou funcional do
dente acometido. São necessários cuidados
especiais, não apenas no que tange à eliminação da
mancha ou restabelecimento da função, mas também na Atualmente, a hipoplasia de esmalte têm sido
considerada como fator de risco à cárie por criar nichos
que favorecem o acúmulo de alimentos (HOFFMANN,
2006). Assim, o autor objetivou investigar a frequência
com que ocorrem essas alterações de esmalte dentário
em diferentes grupos populacionais, bem como procurar
possíveis associações com a ocorrência de cárie
dentária. Concluiu que, na população estudada (624 pré-
escolares de 5 anos e 309 escolares de 12 anos), a
prevalência de hipoplasias e opacidades demarcadas
estão associadas com experiência de cárie dentária e
isso provavelmente ocorre devido à menor resistência REVISTA UNIARA, v.13, n.2, dezembro 2010 140 Amelogênese imperfeita, hipoplasia de esmalte... OBJETIVOS consideração e respeito durante o planejamento e
execução do tratamento, já que são sérias as
consequências emocionais e sociais a que está
submetido o paciente que se encontra com tal prejuízo
funcional e estético (PINHEIRO, 2003). severidade e a distribuição da fluorose dependem da
concentração e duração da exposição do flúor, do
estágio de atividade do ameloblasto e da
suscetibilidade individual, atingindo de maneira mais
severa os dentes permanentes do que os decíduos,
quando ambos estão expostos à mesma concentração
de flúor (CAMPOS, 2009). Os defeitos do esmalte apresentam etiologia diversa
ou causas associadas (BEZERRA, 2006). Em estudo
realizado pelo autor, dois casos clínicos foram
apresentados. No primeiro, a criança apresentava
opacidades definidas, de cor creme, nas superfícies
vestibulares dos incisivos centrais permanentes,
superiores e inferiores. Apresentava também opacidades
difusas envolvendo todas as superfícies dos molares
permanentes. No segundo caso clínico, apresentava
opacidades difusas no terço incisal e oclusal de todos
os dentes permanentes. No primeiro caso, a presença
de opacidades pode ser atribuída aos eventos de
infecções recorrentes que acometeram a criança nos 2
primeiros anos de vida, época em que está ocorrendo a
amelogênese. No segundo caso, o paciente era portador
de fibrose cística. Especula-se que esses pacientes
possam desenvolver defeitos de esmalte, já que a
patologia está presente desde o início de suas vidas,
coincidindo com a amelogênese dos dentes permanentes. Em ambos os casos, o tratamento consistiu em orientação
de higiene oral e dieta. Concluíram que é de fundamental
importância que o clínico saiba reconhecer essas
anomalias para que possa fornecer orientações corretas
ao seu paciente. A fluorose dental ocorre devido aos efeitos do flúor
sobre os ameloblastos, alterando a nucleação e o
crescimento dos cristais de hidroxiapatita, formando
um esmalte defeituoso. Além disso, o flúor interfere
também no metabolismo do cálcio (GUEDES-
PINTO, 1999). ,
)
Para facilitar o diagnóstico da fluorose, algumas
classificações foram propostas. Thylstrup & Fejerskov
(1978) propuseram um sistema de classificação, que
consta de dez pontuações destinadas a caracterizar o
grau macroscópico de fluorose e suas correlações com
as alterações histológicas. No Grau 0 a translucidez
normal do esmalte permanece após prolongada
secagem. No Grau 1, há indiferentes linhas brancas. No Grau 2 as superfícies lisas apresentam linhas mais
pronunciadas de opacidade, podendo ter,
ocasionalmente, confluência de linhas adjacentes, e as
superfícies oclusais apresentam zonas dispersas de
opacidade menor que 2 mm de diâmetro e opacidade
acentuada nos cumes das cúspides. OBJETIVOS No Grau 3 as
superfícies lisas apresentam mesclas de irregulares
áreas nubladas de opacidade e as superfícies oclusais,
áreas confluentes de opacidade marcadas que parecem
quase normais, mas geralmente são circunscritas por
uma borda de esmalte opaco. No Grau 4 toda a
superfície lisa possui exposições marcadas de
opacidade ou aparência de giz branco, com partes da
superfície exposta ao atrito parecendo menos afetadas,
e as superfícies oclusais apresentam-se inteiras com
exposições marcadas de opacidade, sendo que o atrito
é muitas vezes pronunciado logo após a erupção. No
Grau 5 as superfícies lisas e oclusais apresentam
opacidade com perda focal do esmalte com 2 mm de
diâmetro. No Grau 6 as superfícies lisas apresentam
estrias regularmente dispostas em faixas horizontais
menores que 2 mm de extensão vertical, e as superfícies A fluorose dental é um distúrbio no desenvolvimento
dentário causado pela presença excessiva e/ou crônica
de fluoreto durante seu período formativo, sendo uma
lesão hipomineralizada, subsuperficial profunda até uma
superfície de esmalte externo, bem mineralizado que,
em casos mais severos, se rompe logo após a erupção
(RIBAS, 2004). A ingestão crônica de fluoreto causa
mudanças macroscópicas, cuja severidade é
proporcional ao grau de porosidade subsuperficial
(CHALUB, 2008; LEITE, 2007; PIRES, 2001;
ZENKNER, 2005). Caracteriza-se clinicamente por
apresentar esmalte opaco e manchas de coloração que
podem variar do branco ao castanho-escuro, como
também apresentar áreas hipoplásicas e de erosão. A REVISTA UNIARA, v.13, n.2, dezembro 2010 141 BEVILACQUA et al. oclusais apresentam áreas confluentes com menos de
3 mm de diâmetro, além de perda de esmalte marcado
por atrito. No Grau 7 as superfícies lisas possuem perda
de esmalte ultraperiféricas, em áreas irregulares
envolvendo menos da metade da superfície inteira, e
as superfícies oclusais apresentam mudanças na
morfologia, causadas pela fusão de poços e desgaste
acentuado. No Grau 8 as superfícies lisas e oclusais
apresentam perda de esmalte ultraperiféricas
envolvendo menos da metade da superfície e, no Grau
9, apresentam perda da parte principal do esmalte com
a mudança na aparência anatômica da superfície dental. pode ser afetada. As manchas marrons são
generalizadas e os dentes frequentemente apresentam
aparência corroída. A fluorose dental, segundo Moysés (2002), pode
ser classificada como leve, moderada, severa e sistêmica. A fluorose leve causa apenas alterações estéticas,
caracterizadas por pigmentação branca do esmalte
dentário. OBJETIVOS A moderada e a severa são caracterizadas por
manchas amarelas ou marrons, além de defeitos
estruturais no esmalte, apresentando repercussões
estéticas, morfológicas e funcionais. A sistêmica é
provocada por ingestão de altas concentrações de flúor
(acima de 8 ppm) e provoca alterações esqueléticas,
articulares, neurológicas e nefrológicas, dentre outras. ç
p
p
Dean (1942) elaborou um índice epidemiológico
com descrição do aspecto clínico do dente
correspondente a cada uma das pontuações
(Classificação de Dean). Os pontos variam de 0 a 5,
correspondendo respectivamente a normal, duvidosa,
muito leve, leve, moderada, grave. Na normal, o
esmalte apresenta a habitual estrutura translúcida,
semivitreforme, com superfície lisa, lustrosa e, em geral,
de cor branca amarelada pálida. Na duvidosa, o
esmalte revela aberrações ligeiras da translucidez do
esmalte normal, variando de elevações brancas a
brancas ocasionais, sendo esta classificação utilizada
nos casos em que nenhum diagnóstico definitivo da
forma mais branda de fluorose, nem uma classificação
do "normal", se justificam. Na muito leve, o esmalte
apresenta pequenas áreas opacas espalhadas
irregularmente sobre o dente, mas não envolvendo mais
do que 25% da superfície dental, sendo frequentemente
incluídos, nesta classificação, os dentes mostrando não
mais que cerca de 1 a 2 mm de opacidade branca na
ponta das cúspides dos pré-molares ou segundos
molares. Na classificação leve, as áreas brancas opacas
no esmalte dos dentes são mais extensas, mas não
envolvem mais do que 50% do dente. Na moderada
todas as superfícies do esmalte dos dentes são afetadas,
e as superfícies estão sujeitas a desgaste por atrito,
frequentemente desfiguradas por mancha castanha. E,
por último, a grave, que inclui dentes anteriormente
classificados como "moderadamente severo e severo",
em que todas as superfícies do esmalte são afetadas e
a hipoplasia é tão acentuada que a forma geral do dente g
g
O diagnóstico precoce e diferencial da fluorose é
essencial para o estabelecimento do correto tratamento. O aumento da incidência da fluorose pode estar
associado ao uso do creme dental fluoretado e,
principalmente, ao consumo de água fluoretada de
abastecimento público (ZENKNER, 2005;
CAMPOS, 2009; MOYSÉS, 2002). Os mecanismos
etiopatogênicos dessa alteração parecem estar
relacionados a um possível efeito antienzimático do
flúor, que dificulta a remoção das proteínas da matriz
do esmalte, interferindo no pleno crescimento de seus
cristais (ZENKNER, 2005). REVISTA UNIARA, v.13, n.2, dezembro 2010 alterações (TAMES, 1998). alterações (TAMES, 1998). Sendo assim, vários estudos buscam estudar a
prevalência, características clínicas e tratamento da
fluorose dentária. Campos et al. (2009) propuseram
avaliar a prevalência de fluorose em escolares de Brasília,
Distrito Federal, comunidade na qual o teor médio de
flúor na água de abastecimento público é de 0,8 ppm. Concluíram que o percentual de crianças livres de
fluorose se apresentou elevado (85,36%), que entre as
crianças examinadas 14,64% apresentaram fluorose
dentária em níveis muito leve a moderado e não houve
diferença entre os sexos quanto à prevalência de fluorose. O tratamento dependerá do tipo de alteração
existente, relacionado com a profundidade e coloração
das manchas. O tratamento para as alterações de
esmalte descritas poderá ser efetivo e eficaz, desde
que o cirurgião-dentista realize anamnese e diagnóstico
diferencial adequados de modo a restabelecer a
harmonia estética e funcional, além de contribuir para
a melhora da autoestima dos pacientes o mais
precocemente possível. Assim deve haver uma
associação entre conhecimento e habilidade do
profissional aos materiais odontológicos de última
geração (RIBAS, 2004; LEITE, 2007). A escolha do
tratamento depende da gravidade das alterações, indo
desde os mais conservadores, como clareamentos,
macroabrasões e microabrasões, até os restauradores
ou protéticos, como restaurações diretas ou indiretas
(LEITE, 2007). Com o objetivo de revisar a literatura a respeito da
fluorose dental e avaliar sua presença em escolares da
rede pública da Cidade de Curitiba, foi verificado que
a fluorose dental não constitui um problema
epidemiológico relevante para a população estudada,
e as iniciativas para o seu controle devem levar em
conta a autopercepção do problema pela própria
população e ainda é de extrema importância o
monitoramento rigoroso dos teores de flúor na água
de abastecimento (MOYSÉS, 2002). O clareamento dental é uma opção conservadora
no tratamento estético, podendo inclusive ser associado
a outros procedimentos como restaurações com resina
composta e facetas laminadas de porcelana. O
clareamento pode ser recomendado nos casos de
manchas por fluorose e tetraciclina, além de traumas
durante a formação do esmalte (TAMES, 1998). No
caso da amelogênese imperfeita, a técnica do
clareamento sozinha não será suficiente para devolver
ao dente a estética e, em casos mais severos, a função,
sendo necessário realizar um procedimento restaurador
adequado. Já para a hipoplasia, tenta-se inicialmente
a técnica de microabrasão; se não funcionar, se opta
por uma restauração (BARATIERI, 2004). OBJETIVOS O diagnóstico da fluorose
dental é complexo e deve envolver exame clínico
cuidadoso, com dentes limpos, secos e iluminados,
tendo em vista a necessária distinção entre esta
patologia e outras alterações do esmalte, além de uma
anamnese cuidadosa, visando à identificação da fonte
dos fluoretos ingeridos em excesso. Mesmo após quase um século de descrição das
desfigurações dentárias causadas pelo fluoreto, ainda
há muito que se compreender a respeito da patologia,
pois são amplas as suas manifestações clínicas, os
mecanismos pelos quais o fluoreto age durante a
amelogênese não são claros e a susceptibilidade
individual é uma variável relevante e de difícil controle,
sendo que as pessoas acometidas pela fluorose
devem receber tratamento odontológico restaurador
para que possam voltar ao convívio social saudável
(PIRES, 2001). REVISTA UNIARA, v.13, n.2, dezembro 2010 142 Amelogênese imperfeita, hipoplasia de esmalte... Amelogênese imperfeita, hipoplasia de esmalte... REVISTA UNIARA, v.13, n.2, dezembro 2010 DISCUSSÃO As manchas em esmalte dental, causadas por
distúrbios de desenvolvimento, apresentam-se como
anomalias de estrutura, podendo afetar as dentições
decídua e permanente (PITHAN, 2002). Dentre eles
estão a amelogênese imperfeita, a hipoplasia de
esmalte, e a fluorose dental. p
Com relação à hipoplasia de esmalte, alguns autores
a definem como uma formação incompleta ou deficiente
da matriz orgânica do esmalte durante a formação do
germe dentário (BRAGA, 2005; PITHAN, 2002), e
outros, como defeitos quantitativos do esmalte
associados a uma menor espessura do mesmo na área
afetada (HOFFMANN, 2007; RUSCHEL, 2006;
BASSO, 2007). Pode afetar ambas as dentições, mas
na dentição decídua não ocorre de maneira tão severa
(BRAGA, 2005; PITHAN, 2002). Segundo Braga et
al. (2005, Hoffmann et al. (2007), Pinheiro et al. (2003), Ruschel et al. (2006) e Basso et al. (2007),
apresenta-se como fossas profundas, sulcos ou
fóssulas horizontais ou verticais, bem como áreas com
ausência parcial ou total de esmalte. Uma vez formado
o dente, não se produz mais o defeito. Isso se deve ao
fato dos ameloblastos deixarem de existir apos a
formação do esmalte (BRAGA, 2005). Vários autores definem a amelogênese imperfeita
como uma alteração de caráter hereditário (BRAGA,
2005; PITHAN, 2002; RUSCHEL, 2006; AUGUSTO,
2005; MELO, 2007; BRUSCO, 2008; BASSO,
2007), sendo também denominada hipoplasia hereditária
(BRAGA, 2005; PITHAN,2002). Segundo Pithan et
al. (2002) e Brusco et al. (2008), a amelogênese
imperfeita não está relacionada a alterações de ordem
geral ou sistêmica. Com relação aos tipos de
amelogênese imperfeita, pode-se citar a hipoplásica,
hipomineralizada e hipomaturada. As manifestações clínicas da amelogênese imperfeita
são variáveis, podendo desencadear impulso lingual,
levando à mordida aberta anterior e aos defeitos
estruturais que levam à diminuição da dimensão vertical
(PITHAN, 2002; AUGUSTO, 2005; MELO, 2007). Os pacientes que apresentam essa anomalia, segundo
alguns autores (PITHAN, 2002; AUGUSTO, 2005;
BRUSCO, 2008), são mais predispostos a problemas
gengivais, devido à aderência e retenção da placa
bacteriana no esmalte. O diagnóstico dessa patologia
dentária deve ser baseado nas características clínicas
e radiográficas de cada caso e no modo de herança,
sendo que, quando realizado precocemente, contribui
para minimizar ou evitar sequelas (BRUSCO, 2008). Quanto ao tratamento, Melo et al. (2007) relatam
que em pacientes jovens deve ser preservada a maior
quantidade de estrutura dental possível, até que a idade As hipoplasias podem ser diferenciadas, de acordo
com o fator etiológico, em dois tipos: sistêmica ou local. alterações (TAMES, 1998). Foi relatado um caso clínico em que duas irmãs
gêmeas dizigóticas apresentavam fluorose dentária em
graus diferentes. A percepção das manchas de fluorose
dentária como causa de comprometimento estético
dependerá do grau de alteração dos dentes e das
concepções de estética de cada indivíduo, seja criança,
pai ou profissional. O odontopediatra deve estar atento
ao desenvolvimento de novas tecnologias para o
tratamento das manchas de fluorose dental; entretanto,
deve-se sempre respeitar a percepção, os anseios e
as expectativas de seus pacientes e suas famílias
(CHALUB, 2008). Anomalia significa que houve uma variação ou
desvio de uma característica ou estrutura anatômica,
podendo ser congênita, genética ou adquirida e, se não
forem diagnosticadas precocemente, podem originar
problemas graves. É importante estudar e conhecer
essas anomalias dentais, pois a detecção precoce pode
alterar significativamente o prognóstico da dentição
afetada, sendo essa a função do médico e do cirurgião-
dentista, tanto o clínico geral quanto o odontopediatra
ou aquele que atua na área de dentística restauradora. O diagnóstico das alterações dentárias de Conforme observado, algumas opções de
tratamento são descritas na literatura com o objetivo
de melhorar a estética dos pacientes. O tratamento do
esmalte alterado consiste em minimizar esses danos
estéticos e é realizado pela remoção de sua camada
superficial até alcançar o esmalte normal. É muito
importante reconhecer clinicamente essa condição,
identificar as suas causas e estabelecer o seu
diagnóstico diferencial, sendo fundamental para a
condução clínica do paciente afetado por essas REVISTA UNIARA, v.13, n.2, dezembro 2010 143 BEVILACQUA et al. leve à necessidade de realização de técnica
restauradora para reabilitar a dentição. Pithan et al. (2002) reforçam ainda que o plano de tratamento deve
ser meticuloso. Brusco et al. (2008) citam que o
tratamento depende da gravidade da desordem, da
idade, nível socioeconômico e da saúde bucal do
paciente, podendo incluir restaurações estéticas,
confecção de coroas de aço ou de resina composta,
placas de restabelecimento de dimensão vertical,
controle de sensibilidade dentinária e orientação de
higiene bucal, sendo a abordagem multiprofissional um
fator fundamental para o sucesso do tratamento. desenvolvimento requer, além de avaliação clínica,
avaliação radiográfica, sendo que o tratamento deve
ser iniciado assim que se reúnam todas as condições
necessárias ao bom encaminhamento do caso clínico
com a finalidade de restabelecer as condições para
um adequado desenvolvimento e crescimento
(SEABRA, 2008). DISCUSSÃO Os fatores etiológicos sistêmicos incluem deficiências
nutricionais, deficiência de vitaminas A, C, D, deficiência
de cálcio (hipocalcemia), fósforo, doenças que
interferem no metabolismo de cálcio, entre elas o
hipotireoidismo, o hipoparatireoidismo e o diabetes
maternos não controlados, radiação X, parto
prematuro, baixo peso ao nascimento, traumatismos
por ocasião do nascimento, sífilis congênita, doenças
exantematosas (como febre exantematosa, sarampo,
varicela, escarlatina, rubéola, hipoxia cerebral e
desnutrição), qualquer doença sistêmica grave, fatores REVISTA UNIARA, v.13, n.2, dezembro 2010 144 Amelogênese imperfeita, hipoplasia de esmalte... clinicamente significativo, ocorre durante o 2.º e o 3.º
anos de vida (PINHEIRO, 2003). Manifestam-se por
opacidades inicialmente dispersas de cor branca, que
se tornam mais marcadas e de cor amarelo-
acastanhado à medida que se tornam mais graves
(RUSHEL, 2006). idiopáticos (desconhecidos), ingestão de medicamentos
(como a tetraciclina, talidomida e anti-inflamatórios não
esteroidais – Aines, derivados do ácido pirazolônico,
que têm o poder de atravessar a barreira placentária e
interferir na formação dos dentes decíduos e
permanentes, ocasionando malformação do esmalte
dental), traumatismos cerebrais e defeitos neurológicos,
distúrbios respiratórios (asma, doença hemolítica do
recém-nascido, asfixia neonatal) e várias manifestações
e tipos de síndromes (BRAGA, 2005; PITHAN, 2002;
PINHEIRO, 2003). A fluorose dental leve causa apenas alterações
estéticas, caracterizadas por pigmentação branca do
esmalte dentário. A fluorose moderada e severa,
caracterizada por manchas amarelas ou marrons, além
de defeitos estruturais no esmalte, apresenta
repercussões estéticas, morfológicas e funcionais
(MOYSÉS, 2002). Já a hipoplasia de esmalte local é mais conhecida
como Dente de Turner (BRAGA, 2005; PITHAN,
2002; PINHEIRO, 2003; RUSHEL, 2006), pois, de
acordo com Braga et al. (2005), Turner foi o 1.° a
descrever a hipoplasia do tipo localizada quando notou
defeitos do esmalte de dois pré-molares e os relacionou
à infecção apical nos molares decíduos mais próximos. São limitadas a dentes isolados (BRAGA, 2005;
PITHAN, 2002), sendo os incisivos superiores e os
pré-molares os de maior prevalência na população
(BRAGA, 2005). Dean, cirurgião-dentista e epidemiologista,
trabalhando nos Estados Unidos, elaborou em 1934 –
e aprimorou em 1942 – um índice de classificação da
fluorose dentária que leva o seu nome, e que se tornou
índice de aplicação mundial, sendo baseado em
variações no aspecto estético do esmalte, desde normal
até a forma grave, abrangendo seis categorias. REFERÊNCIAS AUGUSTO, L. et al. Amelogênese imperfeita. RGO, v.53, n.3, p. 251-54, jul/ago/set. 2005. BARATIERI, L.N. Caderno de Dentística:
Clareamento Dental. 1 ed. São Paulo: Santos,
2004. 129p. Por ser difícil o diagnóstico diferencial entre esses
tipos de alterações dentais, é importante que mais
estudos sejam realizados para que se possa conhecer
mais a respeito de cada uma das patologias do esmalte,
facilitando o diagnóstico e, conseqüentemente,
possibilitando a escolha adequada do tratamento a ser
empregado. BASSO, A.P. et al. Hipomineralização molar –
incisivo. Rev Odonto Ciênc, v.22, n.58, p.371-76,
out/dez. 2007. BENDO, C.B. et al. Hipoplasia de esmalte em
incisivos permanentes: um acompanhamento de 6
meses. RGO, v.55, n.1, p.107-12, jan/mar. 2007. DISCUSSÃO Contudo,
havia limitações importantes nesse sistema de
classificação, como o fato de as manchas marrons, ou
acastanhadas – adquiridas pós-eruptivamente – não
refletirem a suposta severidade da lesão, mas a
capacidade de pigmentação de uma superfície de esmalte
mais poroso, a qual é também inerente ao meio bucal. As opções de tratamento citadas na literatura
incluem aplicação tópica de fluoreto, em casos de
sensibilidade, além de microabrasão do esmalte,
clareamento dental, restaurações diretas e indiretas em
resina composta ou cerâmica, coroas totais
metalocerâmicas (BRAGA, 2005; PITHAN, 2002;
PINHEIRO, 2003). O tratamento restaurador é
necessário quando há presença de cavidades cariosas,
risco de futuras lesões, comprometimento estético e
sensibilidade dentinária (BRAGA, 2005). Assim, Thylstrup & Fejerskov propuseram, em 1978,
um índice que classifica as lesões fluoróticas em dez graus
crescentes de gravidade (0 – 9), de acordo com sua
aparência clínica. No entanto, ao contrário do índice de
Dean, este índice (conhecido como TF – Thylstrup/
Fejerskov) preconiza a secagem da superfície dentária
antes do exame. Tal fato está de acordo com os atuais
conhecimentos sobre as características histológicas do
esmalte fluorótico. Assim, no índice TF, ao se examinar
o esmalte seco, tende-se a avaliar o aspecto real da
superfície em questão. Além disto, propõe cinco
classificações para os casos mais severos da doença,
que no índice de Dean são apenas classificados como
"grave" (ZENKNER, 2005; THYLSTRUP &
FEJERSKOV, 1978; DEAN, 1942). A fluorose dentária é definida por alguns autores
como um distúrbio no desenvolvimento dentário
causado pela presença excessiva e/ou crônica de
fluoreto durante seu período formativo (PINHEIRO,
2003; RUSHEL, 2006; PIRES, 2001; MOYSÉS,
2002), também como um tipo de hipoplasia de esmalte
(PITHAN, 2002; ZENKNER, 2005) causada pela
ingestão excessiva de flúor a partir de diversas fontes,
como pastas de dente para adultos reforçadas com
flúor, suplementos de flúor, sucos de frutas, água
fluoretada e medicamentos infantis. O período crítico, Alguns autores citaram as opacidades como um tipo
de fluorose dentária (HOFFMANN, 2007; RUSHEL, 145 REVISTA UNIARA, v.13, n.2, dezembro 2010 BEVILACQUA et al. BEVILACQUA et al. do germe dentário durante seu desenvolvimento. O
tratamento é restabelecer a anatomia dental e a harmonia
entre oclusão, função e estética. 2006; BASSO, 2007). A opacidade difusa também é
uma anormalidade envolvendo uma alteração na
translucidez do esmalte, de grau e coloração variáveis,
como a opacidade demarcada. DISCUSSÃO Todavia, não existe um
limite claro entre o esmalte normal adjacente e a
opacidade difusa, podendo apresentar-se clinicamente
de forma linear ou em placas, ou ter uma distribuição
confluente. do germe dentário durante seu desenvolvimento. O
tratamento é restabelecer a anatomia dental e a harmonia
entre oclusão, função e estética. 3) A fluorose dentária é uma hipoplasia de esmalte
causada pela ingestão crônica de flúor. Manifesta-se
por opacidades inicialmente dispersas de cor branca,
que se tornam mais marcadas e de cor amarelo-
acastanhado à medida que se tornam mais graves. O
tratamento depende do grau de escurecimento dental,
podendo ser mais conservador, como o clareamento,
ou mais invasivo, como a confecção de facetas. O tratamento do esmalte acometido por fluorose é
realizado com finalidade eminentemente estética, através
da remoção de sua camada superficial até alcançar o
esmalte normal. Nos casos mais brandos, a técnica da
microabrasão do esmalte pode ser empregada com
êxito, podendo-se utilizar o ácido fosfórico a 37%
associado à pedra-pomes, que oferece como vantagem
um menor custo e desgaste de esmalte clinicamente
insignificante, além de constituir-se um método de rápida
execução, pois não requer anestesia, e com um alto grau
de satisfação pelo paciente, já que não existem relatos
de recorrência das manchas ou perda de vitalidade dos
dentes tratados. Já os casos mais severos requerem
confecção de facetas de resina composta ou coroas
protéticas (PINHEIRO, 2003; ZENKNER, 2005). O tratamento do esmalte acometido por fluorose é
realizado com finalidade eminentemente estética, através
da remoção de sua camada superficial até alcançar o
esmalte normal. Nos casos mais brandos, a técnica da
microabrasão do esmalte pode ser empregada com
êxito, podendo-se utilizar o ácido fosfórico a 37%
associado à pedra-pomes, que oferece como vantagem
um menor custo e desgaste de esmalte clinicamente
insignificante, além de constituir-se um método de rápida
execução, pois não requer anestesia, e com um alto grau
de satisfação pelo paciente, já que não existem relatos
de recorrência das manchas ou perda de vitalidade dos
dentes tratados. Já os casos mais severos requerem
confecção de facetas de resina composta ou coroas
protéticas (PINHEIRO, 2003; ZENKNER, 2005). 4) Para estabelecer o diagnóstico diferencial e,
consequentemente, o melhor tratamento para o caso,
é imprescindível o conhecimento das características
correspondentes a cada uma dessas alterações, a
realização de uma excelente anamnese, exame físico,
e em alguns casos, exame radiográfico. CONCLUSÃO De acordo com a revisão de literatura realizada,
pode-se concluir que: 1) A amelogênese imperfeita é um tipo de hipoplasia
de esmalte hereditária, em que o portador não apresenta
nenhuma alteração sistêmica. Os tratamentos geralmente
são complexos e demorados, dependendo da gravidade
da alteração. BEZERRA, A.C.B., AZEVEDO, T.D.P., TOLEDO,
O.A. Defeitos de desenvolvimento do esmalte
dentário: relato de casos clínicos. Rev Assoc Paul
Cir Dent, v.60, n.4, p.265-70. 2006. BRAGA, L.C.C. et al. Hipoplasia de esmalte
localizada - Dente de Turner. RGO, v.53, n.4,
p.329-34, out/nov/dez. 2005. BEZERRA, A.C.B., AZEVEDO, T.D.P., TOLEDO,
O.A. Defeitos de desenvolvimento do esmalte
dentário: relato de casos clínicos. Rev Assoc Paul
Cir Dent, v.60, n.4, p.265-70. 2006. 2) A hipoplasia de esmalte é um defeito quantitativo
do esmalte associado a uma menor espessura do mesmo
na área afetada. Este defeito é causado por algum
estímulo genético ou ambiental sobre os ameloblastos BRAGA, L.C.C. et al. Hipoplasia de esmalte
localizada - Dente de Turner. RGO, v.53, n.4,
p.329-34, out/nov/dez. 2005. REVISTA UNIARA, v.13, n.2, dezembro 2010 146 Amelogênese imperfeita, hipoplasia de esmalte... CYPRIANO,S. Prevalência de defeitos de esmalte
e sua relação com cárie dentária nas dentições
decídua e permanente, Indaiatuba, São Paulo, Brasil. Cad. Saúde Pública, Rio de Janeiro, v.23, n.2,
p.435-44, fev. 2007. CYPRIANO,S. Prevalência de defeitos de esmalte
e sua relação com cárie dentária nas dentições
decídua e permanente, Indaiatuba, São Paulo, Brasil. Cad. Saúde Pública, Rio de Janeiro, v.23, n.2,
p.435-44, fev. 2007. BRUSCO, L.C. et al. Amelogênese imperfeita –
cinco anos de acompanhamento. RFO, v.13; n.1,
p.59-63, jan/abr. 2008. CAMPOS, D.L.; FARIAS, D.G.; TOLEDO, O.A. Prevalência de fluorose dentária em escolares de
Brasília – Distrito Federal. Rev Odontol Univ São
Paulo, v.12, n.3, p.225-30, Jul. 1998. Disponível em
http://www.scielo.br/scielo. Acesso em 15 set. 2009 LEITE, P.S.A. et al. Microabrasão de esmalte:
solução estética para dentes com fluorose. Rev
Assoc Paul Cir Dent, v.61, n.2, p.139-42. 2007. CHALUB, L.L.F.; MARTINS, C.C.; PAIVA, S.M. Percepção estética das manchas de fluorose
dentária: relato de caso de gêmeas dizigóticas. Rev
Odonto Ciênc, v.23, n.3, p.302-6. 2008. MACEDO, L.A. et al. Defeitos de esmalte em
dentes decíduos e permanentes em crianças
prematuras e entubadas. Horizonte Científico,
Uberlândia, v.2, p.1-24. 2003. DEAN, H.T. The investigation of physiological
effects by the epidemiological method. In:
MOULTON, R.F., ed. Fluorine and dental health. Washington, DC, American Association for the
Advancement of Science, 1942. DEAN, H.T. RECEBIDO EM 27/10/2010
ACEITO EM 4/12/2010 CONCLUSÃO The investigation of physiological
effects by the epidemiological method. In:
MOULTON, R.F., ed. Fluorine and dental health. MACHADO, M.A.A.M.; TELLES, P.D.S.; SILVA,
S.M.B. Diagnostico de manchas do esmalte. Rev
Assoc Paul Cir Dent, v.55, n.3, p.206-09. Ilus,
mai/jun. 2001. Washington, DC, American Association for the
Advancement of Science, 1942. MELO, T.A.F. et al. Amelogênese imperfeita –
relato de caso. Revista Dentística on line, v.7,
n.16, p.104-09, jul/dez. 2007. GERLACH, R. F.; SOUSA, M. da L. R. de;
CURY, J.A. Esmalte Dental com Defeitos: de
marcador biológico a implicações clínicas. Rev. Odonto Ciência, Porto Alegre, v.15, n.31, p.87-
102. 2000. MOYSÉS, S.J. et al. Fluorose dental: ficção
epidemiológica?. Rev Panam Salud Publica/Pan
Am J Public Health, v.12, n.5, p. 339-46. 2002. GUEDES-PINTO, A. et al. Reabilitação Bucal
em Odontopediatria: atendimento integral. São
Paulo: Santos, 1999. 320p. OLIVEIRA, A.F.B. & ROSENBLAT, A. Defeitos
do esmalte: o que o odontopediatra precisa saber. Rev ABO Nac, v.10, n.5, p.274-77, out/nov. 2002. HANSER-DUCATTI, C. et al. Relação entre
estado nutricional e alterações do esmalte dental em
escolares de Botucatu-SP. Cienc Odontol Brás,
v.7, n.1, p.84-9, jan/mar. 2004. PINHEIRO, I.V.A. et al. Lesões brancas no esmalte
dentário: como diferenciá-las e tratá-las. Rev Bras
Patol Oral, v.2, n.1, p.11-18.ilus, jan/mar. 2003. PIRES, M.B.O. Fluorose dentária endêmica: revisão
da literatura. Unimontes Científica, v.2, n.2, p.1-
15, set. 2001. HOFFMANN, R.H.S. Defeitos de esmalte nas
dentições decíduas e permanentes. Tese de
Mestrado (Odontologia) – Faculdade de
Odontologia de Piracicaba, da Universidade
Estadual de Campinas, Piracicaba, 2006. PITHAN, J.C.A et al. Amelogênese imperfeita:
revisão de literatura e relato de caso clínico. Rev
ABO Nac, v.10, n.2, p.88-92, abr/mai. 2002. 147 REVISTA UNIARA, v.13, n.2, dezembro 2010 BEVILACQUA et al. RIBAS, A.O. & CZLUSNIAK, G.D. Anomalias do
esmalte dental: etiologia, diagnóstico e tratamento. Publ UEPG Ci Biol Saúde, v.10, n.1, p.23-36,
mar. 2004. TAMES, D. et al. Alterações do esmalte dental
submetido ao tratamento com peróxido de
carbamida 10%. Rev Assoc Paul Cir Dent, v.52,
n.2, p.145-49, mar/abr. 1998. RUSCHEL, H.C. et al. Hipoplasia e
hipocalcificação de primeiros molares permanentes. Rev ABO Nac, v.14, n.2, p.89-94, abr/mai. 2006. SEABRA, M. et al. A importância das anomalias
dentárias de desenvolvimento. Acta Pediatr Port,
v.39, n.5, p.195-00. 2008. SHAFER, W. G.; HINE, M. K.; LEVY, B. M. Distúrbios do Desenvolvimento das Estruturas
Bucais e Parabucais. In:___________. Tratado de
Patologia Bucal. 4. ed. Rio de Janeiro: Guanabara
Koogan, 1987. cap.1, p. 2-79. RUSCHEL, H.C. CONCLUSÃO et al. Hipoplasia e
hipocalcificação de primeiros molares permanentes. Rev ABO Nac, v.14, n.2, p.89-94, abr/mai. 2006. RUSCHEL, H.C. et al. Hipoplasia e
hipocalcificação de primeiros molares permanentes. Rev ABO Nac, v.14, n.2, p.89-94, abr/mai. 2006. SEABRA, M. et al. A importância das anomalias
dentárias de desenvolvimento. Acta Pediatr Port,
v.39, n.5, p.195-00. 2008. SHAFER, W. G.; HINE, M. K.; LEVY, B. M. Distúrbios do Desenvolvimento das Estruturas
Bucais e Parabucais. In:___________. Tratado de
Patologia Bucal. 4. ed. Rio de Janeiro: Guanabara
Koogan 1987 cap 1 p 2 79 THYLSTRUP, A. & FEJERSKOV, O. Clinical
appearance and surface distribution of dental
fluorosis in permanent teeth in relation to histological
changes. Community dentistry and oral
epidemiology, v.6, p.315. 1978. SEABRA, M. et al. A importância das anomalias
dentárias de desenvolvimento. Acta Pediatr Port,
v.39, n.5, p.195-00. 2008. WATANABE, A.S. et al. Restauração de dilaceração
coronária e hipoplasia de esmalte. J Bras Clin
Odontol Int, v.7, n.38, p.126-30, mar/abr. 2003. SHAFER, W. G.; HINE, M. K.; LEVY, B. M. Distúrbios do Desenvolvimento das Estruturas
Bucais e Parabucais. In:___________. Tratado de
Patologia Bucal. 4. ed. Rio de Janeiro: Guanabara
Koogan, 1987. cap.1, p. 2-79. SHAFER, W. G.; HINE, M. K.; LEVY, B. M. Distúrbios do Desenvolvimento das Estruturas
Bucais e Parabucais. In:___________. Tratado de
Patologia Bucal. 4. ed. Rio de Janeiro: Guanabara
Koogan, 1987. cap.1, p. 2-79. ZENKNER, J.E.A. et al. Fluorose dental: aspectos
históricos, etiopatogênicos e clínicos. Rev. Saúde,
v.31, n.1-2, p.34-41. 2005. REVISTA UNIARA, v.13, n.2, dezembro 2010 148
|
https://openalex.org/W2161394576
|
https://europepmc.org/articles/pmc2949756?pdf=render
|
English
| null |
Motif prediction to distinguish LPS-stimulated pro-inflammatory vs. antibacterial macrophage genes
|
Immunome research
| 2,010
|
cc-by
| 8,028
|
RESEARCH Open Access Open Access © 2010 Kollipara and Perumal; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative
Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly cited. Motif prediction to distinguish LPS-stimulated
pro-inflammatory vs. antibacterial macrophage
genes Rahul K Kollipara, Narayanan B Perumal* Abstract Background: Innate immunity is the first line of defence offered by host cells to infections. Macrophage cells
involved in innate immunity are stimulated by lipopolysaccharide (LPS), found on bacterial cell surface, to express a
complex array of gene products. Persistent LPS stimulation makes a macrophage tolerant to LPS with down
regulation of inflammatory genes (“pro-inflammatory”) while continually expressing genes to fight the bacterial
infection (“antibacterial”). Interactions of transcription factors (TF) at their cognate TF binding sites (TFBS) on the
expressed genes are important in transcriptional regulatory networks that control these pro-inflammatory and
antibacterial expression paradigms involved in LPS stimulation. Results: We used differential expression patterns in a public domain microarray data set from LPS-stimulated
macrophages to identify 228 pro-inflammatory and 18 antibacterial genes. Employing three different motif search
tools, we predicted respectively four and one statistically significant TF-TFBS interactions from the pro-inflammatory
and antibacterial gene sets. The biological literature was utilized to identify target genes for the four pro-
inflammatory profile TFs predicted from the three tools, and 18 of these target genes were observed to follow the
pro-inflammatory expression pattern in the original microarray data. Conclusions: Our analysis distinguished pro-inflammatory vs. antibacterial transcriptomic signatures that classified
their respective gene expression patterns and the corresponding TF-TFBS interactions in LPS-stimulated
macrophages. By doing so, this study has attempted to characterize the temporal differences in gene expression
associated with LPS tolerance, a major immune phenomenon implicated in various pathological disorders. and prompt an intracellular signalling cascade which
ends in induction of pro-inflammatory cytokines, che-
mokines, type I interferons, and antimicrobial effectors
that are essential for providing continuous protection
from infection. In highly evolved organisms (starting
from jawed vertebrates) innate immunity is required for
priming adaptive immunity, the second arm of the
immune system with long-term and specific immune
response [2]. Macrophages are immune cells highly
involved in conducting innate immune responsibilities
and also play a role as an antigen-presenting cell (APC)
to T lymphocytes (in adaptive immunity); they express
various TLR genes to mediate the pro-inflammatory and
antimicrobial responses. * Correspondence: nperumal@iupui.edu
School of Informatics, Indiana University-Purdue University Indianapolis,
Indianapolis, Indiana 46202, USA Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 IMMUNOME RESEARCH RESEARCH Classification of antibacterial and pro-inflammatory
macrophage genes In order to distinguish macrophage genes responsible for
antibacterial versus pro-inflammatory phenotypes upon
induction of LPS tolerance, we filtered the differentially
expressed genes reported in Foster et al [6] as shown in
the top part of Figure 1. These authors have performed
two replicates of microarray hybridizations for each of the
following experimental conditions: The naïve (N) state
that corresponds to no LPS treatment, a 4-hour treatment
of LPS to naïve cells (labelled N+L) and a 24-hour LPS
treatment of naïve cells followed by an additional 4-hour
treatment (labelled T+L). Upon the first LPS treatment for
24 hours, the macrophages in the naïve state become tol-
erant and a second treatment for an additional 4 hours
should reduce inflammatory gene expression but sustain
or increase antibacterial gene expression. We employed a
novel filtering approach to distinguish the Foster et al
microarray genes into the pro-inflammatory and antibac-
terial classes. Essentially if a gene’s expression level is X in
N, for it to be a pro-inflammatory gene its expression in N
+L, Y should be greater than X (Y > X) but expression in
T+L, Z should be less than Y (Z < Y). On the other hand
if a gene is expressed higher in N+L compared to N (i.e., Y
> X) and is same or increased in T+L compared to N+L,
(i.e., Z≥Y although we filtered only when Z > Y), then it
was classified as antibacterial. This filtering paradigm is
based on the rationale that while pro-inflammatory genes
are down regulated in the tolerant state, antibacterial
genes are required for infection control. We obtained 228
pro-inflammatory and 18 antibacterial genes (p < 0.05 sig-
nificance) from an annotated set of 45,101 probes in the
microarray indicating the stringency in our phenotypic
restrictions (Additional file 1, Table S1). This categoriza-
tion is further confirmed when we clustered the original
gene expression data of a mixture of these genes (228 + 18
= 246) using the MeV software [11]. As expected, the two
classes of genes clearly separated into two clusters (Figure
2) with the pro-inflammatory set showing down regulation
and antibacterial set showing up-regulation compared to
the tolerant state (T+L/N+L avg). Transcriptional regulation is a crucial biological
mechanism controlling gene regulation in the tolerant
phenotype vs. the basal state that corresponds to no LPS
stimulation. Background Innate immunity, one of the two arms of the immune
system, provides the first line of defence against patho-
gens in mammals and nearly all other living things. Ani-
mals in the lower evolutionary scale, such as insects,
fight off infections solely employing innate immune
mechanisms. The innate immune system quickly alerts
the host of the presence of microbial pathogens and this
response is mediated through the expression of a limited
number of receptors called pattern recognition receptors
(PRRs) to identify pathogen associated molecular pat-
terns (PAMPs) expressed by invading pathogens. Infec-
tions trigger PRRs, such as toll-like receptor (TLR)
genes [1], to recognize PAMPs of invading pathogens LPS, a major component of gram-negative bacterial
cell surface, is a potent stimulator of macrophages. LPS
acts via the TLR4 receptor to trigger downstream Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Page 2 of 12 signalling and expression of pro-inflammatory and anti-
bacterial genes [1]. This induction needs to be under
tight control since dysregulated inflammation can cause
a number of pathological disorders such as septic shock,
autoimmunity, atherosclerosis and cancer [3]. Various
mechanisms of negative regulation of TLR-induced gene
expression have been proposed to dampen uncontrolla-
ble inflammation [4] and these collectively lead to the
phenomenon of “LPS tolerance” [5] wherein there is
decreased expression of pro-inflammatory genes when
there is prolonged LPS administration. Foster et al [6]
have characterized the gene expression profiles of
macrophages differentially treated with LPS to classify
the genes into various phenotypic states including a tol-
erant state obtained by an initial LPS treatment. Their
analysis of the genes expressed in the tolerant pheno-
type categorized the genes as belonging to “tolerizable”
or “non-tolerizable” sets depending on no induction or
further induction respectively during a second LPS
treatment compared to the first one. Although LPS tol-
erance could prevent pathological inflammatory condi-
tions in chronic bacterial infections, there is a strong
need for a persistent antibacterial response to keep the
infections under control. The set of genes that exhibit
the tolerizable phenotype can be considered “pro-
inflammatory” while those belonging to the non-toleriz-
able phenotype as “anti-bacterial”. expression depending on the well-accepted premise of
coordinate expression corresponding to appropriate TF-
TFBS interactions. Further, we show that some of these
TF-TFBS interactions predicted from our analysis have
been biologically validated as transcriptional targets in
the literature with evidence of roles in LPS tolerance. Classification of antibacterial and pro-inflammatory
macrophage genes A number of studies have looked at the
transcriptional programs in LPS-mediated macrophage
stimulation including the roles of TFs in prolonged LPS
treatment [4,7-9]. Roach et al [7] carried out a holistic
approach to identify 92 TFs in human macrophages sti-
mulated with LPS; however, this study did not distin-
guish between the tolerant and basal states since there
was no re-stimulation with LPS in this study. A recent
study characterized the Cebpd TF as a potential regula-
tor of a switch between the basal and tolerant state [9]. A microarray analysis performed by Mages et al [10]
similar to the Foster et al [6] experiment employed in
our study, observed diminished gene expression of a
vast majority of LPS-induced genes upon a second LPS
treatment (tolerizable). However, these authors did not
characterize any of the TF-target interactions responsi-
ble for the various phenotypes distinguished in their
analysis. In order to achieve a global perspective on the
transcriptional regulatory mechanisms inherent in LPS
tolerance, we examined the Foster et al microarray data
[6] for the control of pro-inflammatory vs. antibacterial
gene expression. Using bioinformatics approaches, we
characterized TF-TFBS interactions that differentiate
LPS-stimulated pro-inflammatory vs. antibacterial gene Gene ontology (GO) analysis of the two gene sets gy
y
g
The pro-inflammatory genes could be classified into 13
categories of GO terms while the smaller antibacterial
gene set mapped only to two GO categories. Figure 3 Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Page 3 of 12 Figure 1 Workflow of experimental analysis to distinguish transcriptomic signatures of pro-inflammatory vs. antibacterial genes. Average gene expression values in untreated macrophages, (N)avg, 4 hrs LPS-treated macrophages, (N+L)avg, and 24 hrs LPS-treated followed by
4 hrs re-treated macrophages, (T+L)avg were calculated. Two sets of genes were classified by the conditions mentioned in Methods. Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Page 4 of 12 Figure 2 Heat map showing the differential expression patterns of LPS-stimulated genes. Navg, (N+L) avg and (T+L) avg are as described in
Figure 1. Nabs was introduced to distinguish the patterns clearly and it is equal to one. The rationale was that the fold change is >1 for both
classes of genes in the N+L phenotype, and <1 for pro-inflammatory and >1 for the antibacterial class genes in the T+L phenotype. Only a
subset of the pro-inflammatory genes is shown. Figure 2 Heat map showing the differential expression patterns of LPS-stimulated genes. Navg, (N+L) avg and (T+L) avg are as described in
Figure 1. Nabs was introduced to distinguish the patterns clearly and it is equal to one. The rationale was that the fold change is >1 for both
classes of genes in the N+L phenotype, and <1 for pro-inflammatory and >1 for the antibacterial class genes in the T+L phenotype. Only a
subset of the pro-inflammatory genes is shown. shows a subset of GO terms from the pro-inflammatory
gene set showing significant enrichment (p < 0.01) in a
number of biologically relevant categories such as
immune response, cytokine and chemokine mediated
signalling pathway, and DNA-dependent regulation of
transcription. expectation-maximization tool that fits a two-compo-
nent finite mixture model to the input sequences for
motif prediction [12]. MotifModeler uses a model selec-
tion approach that best fits a set of motifs to gene
expression values (both up and down regulated) in co-
ordinately expressed genes [13]. PASTAA detects TFBS
based on the prediction of binding affinities of a TF to
promoters and their association with tissue specific
expression of corresponding genes [14]. To reduce false
positives, we considered only the top 70% predicted
TFBS from each tool. Gene ontology (GO) analysis of the two gene sets MEME, MotifModeler and PAS-
TAA individually identified 17, 301 and 350 motifs
respectively from the pro-inflammatory genes while four
motifs were found to be common to all three tools (Fig-
ure 4A). Figure 4B lists the TFBS common to all tools
found in the pro-inflammatory gene set and the Characterization of TFBS in the two data sets We
compared scores from the three TFBS prediction tools
for the four motifs identified in our pro-inflammatory
data set, and similarly predicted from a random set of
228 genes. As shown in Table 1, the scores of MotifMo-
deler and PASTAA from the pro-inflammatory gene set
are significantly higher than the scores from the random
gene data indicating genuine enrichment of these motifs
in our genes with a potential for transcriptional control
of pro-inflammatory specific gene expression. MEME
did not predict any of these four motifs in the random
gene set. The two motifs predicted from PASTAA and
MotifModeler when a similar analysis as described
above was performed on the random gene set showed
reciprocal higher (PASTAA) or relatively similar (Motif-
Modeler) scores for the random genes compared to the pro-inflammatory genes (Table 2) stressing the specifi-
city of the four motifs (Figure 4B, and Tables 1 &2) for
regulating pro-inflammatory gene expression. In the antibacterial gene set, the tools found 22, 7, and
178 motifs respectively with the intersection containing
only one motif (Prrx2 as the profile TF). Due to the lim-
ited size of this latter data set, we did not compare pre-
diction scores for this data with a random data set, as
we did for the pro-inflammatory data. However, we per-
formed target identification for Prrx2 using literature
sources (next section). Characterization of TFBS in the two data sets In order to correlate gene expression with transcrip-
tional regulation, we set out to identify characteristic
TF-TFBS interactions unique to the two classes of
genes. We approached this by employing three indepen-
dent TFBS motif detection tools with different algorith-
mic paradigms and to arrive at a list of predicted TFBS
common to the three tools viz., MEME, MotifModeler
and PASTAA (Middle part of Figure 1). MEME is an Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Page 5 of 12 Figure 3 GO analysis of the pro-inflammatory genes. GO analysis of the pro-inflammatory genes based on annotated biological process GO
terms was done using GOstat [37]. Enrichment of the pro-inflammatory gene set compared to a Mouse Genome Informatics (MGI) murine
genome background d for each GO term is shown with significant p-values (< 0.01). Figure 3 GO analysis of the pro-inflammatory genes. GO analysis of the pro-inflammatory genes based on annotated biological process GO
terms was done using GOstat [37]. Enrichment of the pro-inflammatory gene set compared to a Mouse Genome Informatics (MGI) murine
genome background d for each GO term is shown with significant p-values (< 0.01). Figure 3 GO analysis of the pro-inflammatory genes. GO analysis of the pro-inflammatory genes based on annotated biological process GO
terms was done using GOstat [37]. Enrichment of the pro-inflammatory gene set compared to a Mouse Genome Informatics (MGI) murine
genome background d for each GO term is shown with significant p-values (< 0.01). corresponding profile TFs from TRANSFAC [15]. We
compared scores from the three TFBS prediction tools
for the four motifs identified in our pro-inflammatory
data set, and similarly predicted from a random set of
228 genes. As shown in Table 1, the scores of MotifMo-
deler and PASTAA from the pro-inflammatory gene set
are significantly higher than the scores from the random
gene data indicating genuine enrichment of these motifs
in our genes with a potential for transcriptional control
of pro-inflammatory specific gene expression. MEME
did not predict any of these four motifs in the random
gene set. The two motifs predicted from PASTAA and
MotifModeler when a similar analysis as described
above was performed on the random gene set showed
reciprocal higher (PASTAA) or relatively similar (Motif-
Modeler) scores for the random genes compared to the corresponding profile TFs from TRANSFAC [15]. Validation of TF-TFBS interactions controlling the pro-
inflammatory genes We approached the validation of the various predicted
TFBS in the pro-inflammatory class primarily by (1) Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Page 6 of 12 Figure 4 Comparison of predicted TFBS from MEME, MotifModeler, and PASTAA. A. Venn diagram showing numbers of predicted motifs
from the three tools for the pro-inflammatory genes. The numbers next to the prediction tool names are the total numbers of top 70%
predicted TFBS for each tool. B. List of common predicted motifs and the corresponding profile TFs. Profile TFs corresponding to these four
common TFBS were obtained from TRANSFAC [15]. Figure 4 Comparison of predicted TFBS from MEME, MotifModeler, and PASTAA. A. Venn diagram showing numbers of predicted motifs
from the three tools for the pro-inflammatory genes. The numbers next to the prediction tool names are the total numbers of top 70%
predicted TFBS for each tool. B. List of common predicted motifs and the corresponding profile TFs. Profile TFs corresponding to these four
common TFBS were obtained from TRANSFAC [15]. relating the profile TFs corresponding to the predicted
motifs with experimentally identified target genes for
those TFs, and then (2) checking the expression patterns
of a subset of these transcriptional target genes in our
microarray data (Bottom part of Figure 1). We manually
characterized the target genes that various TFs act on
by using the TRANSFAC database [15] and the Ingenu-
ity Pathway Analysis tool [16]. Table 3 lists the pro-
inflammatory profile TFs identified from the three TFBS
prediction tools and their corresponding biologically
validated target genes. A significant number of these
genes have been implicated in the inflammatory
response including Irf1, Irf2, Ifnb1, Tlr3, Stat1, Stat3,
and Nfkb1 with a number of them regulated in LPS-sti-
mulated macrophage activation [8]. To check for the expression patterns of these identified target genes
within our data set, we filtered the microarray data by
looking for two-fold induction in the N+L set compared
to the N samples plus two-fold repression in the T+L
samples compared to the N+L set. This filtering is simi-
lar to what is done earlier (Figure 1) except for the cut-
offs being two-fold with no p-value threshold here; it
resulted in an expression signature of 2283 genes corre-
sponding to the pro-inflammatory phenotype. Score from random gene set
Blank cells represent TFBS that were not identified by the respective tool in the test or random set. For MotifModeler, higher the TCS score better the prediction
whereas for PASTAA, lower the P-value better the prediction. * Score from the test gene set ** Score from random gene set Blank cells represent TFBS that were not identified by the respective tool in the test or random set. For MotifModeler, higher th
whereas for PASTAA, lower the P-value better the prediction. Validation of TF-TFBS interactions controlling the pro-
inflammatory genes Interestingly, there were 18 genes from this larger pro-
inflammatory gene set that overlapped with the target
genes in Table 3 (highlighted). From the original pro-
inflammatory gene set (of 228 genes), there were only
four that overlapped with the target genes mainly due to
the more stringent p-value filter that was imposed Table 1 Comparison of TFBS prediction scores between test vs. random gene sets
Matrix Accession Number
TF name
MEME
MEME
Motif Modeler
Motif Modeler
PASTAA
PASTAA
E-value*
E-value**
TCS score*
TCS score**
P-value*
P-value**
M01014
SOX
1.60E+07
-
1.49E-02
7.06E-03
2.99E-01
7.38E-01
M00699
IRF-8
6.90E-01
-
1.50E-02
-
3.50E-05
9.51E-01
M00233
MEF-2
1.60E+07
-
1.54E-02
6.91E-03
5.75E-01
7.68E-01
M00062
IRF-1
2.60E-07
-
1.55E-02
6.86E-03
7.40E-05
- Table 1 Comparison of TFBS prediction scores between test vs. random gene sets Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Page 7 of 12 Table 2 Reciprocal comparison of TFBS prediction scores between random vs. test gene sets
Matrix Accession Number
TF name
MEME
MEME
Motif Modeler
Motif Modeler
PASTAA
PASTAA
E-value*
E-value**
TCS score*
TCS score**
P-value*
P-value**
M00649
MAZ
-
4.10E+05
1.73E-02
8.28E-03
2.49E-02
6.92E-03
M00423
Foxj2
-
1.80E+07
1.45E-02
7.25E-03
6.00E-01
5.05E-01
* Score from the test gene set
** Score from random gene set
Blank cells represent TFBS that were not identified by the respective tool in the test or random set. For MotifModeler, higher the TCS score better the prediction
whereas for PASTAA, lower the P-value better the prediction. mparison of TFBS prediction scores between random vs. test gene sets Table 2 Reciprocal comparison of TFBS prediction scores between random vs. test gene sets et
t were not identified by the respective tool in the test or random set. For MotifModeler, higher the TCS score better the prediction
e P-value better the prediction. LPS re-stimulation, we analyzed the expression levels of
our 18 target genes in the Mages et al [10] data available
in the public domain. Although these authors have
added the tolerant (T) stage to their analysis (compared
to the Foster et al data in our analysis), we could still
look for gene expression patterns specific to the pro-
inflammatory phenotype similar to our study. Profile TFs corresponding to the predicted motifs are searched against the TRANSFAC [14] and IPA [15] databases and the various target genes known to be
associated with these profile TFs are listed. All profile TFs for each motif (Matrix Accession Number) in the databases are shown. * Highlighted genes showed expression pattern as in Figure 5. Validation of TF-TFBS interactions controlling the pro-
inflammatory genes Essen-
tially, we compared the fold change between the N and
N+L stages [(N+L)/N] in their data with that between
the T and T+L stages [(T+L)/T] with the former condi-
tion setting up the tolerant (T) phenotype and the latter
showing differential gene expression of ‘tolerizable’ and
‘non-tolerizable’ genes. Almost all of our 18 transcrip-
tional targets (17/18 with Isl1 being the exception)
showed the pro-inflammatory expression pattern of
being repressed (tolerizable) upon the second LPS treat-
ment (Additional file 4, Table S4). earlier. Next, we investigated the expression patterns of
these 18 target genes in our microarray data. As
expected, all 18 genes showed the pro-inflammatory
phenotype (Figure 5) (p = 2.94 × 10-4). We also sampled
10 random sets of 18 genes each (generated using the
RSAT tool [17]) for the pro-inflammatory pattern of
expression in our microarray data (Additional file 2,
Table S2) and found either no or very few genes (0, 1 or
2 out of 18) showing the pattern (p = 0.897). This target
gene analysis clearly establishes our predictions as
potentially genuine, biologically meaningful TF-TFBS
interactions for the pro-inflammatory phenotype. A
similar analysis on the two motifs (and the correspond-
ing profile TFs, Maz and Foxj2, Table 2) from the origi-
nal 228 random genes used in the TFBS prediction
algorithms, identified a different set of 18 target genes
(Additional file 3, Table S3) none of which had the pro-
inflammatory expression pattern. As a further step to
verify the role of these validated TF-TFBS interactions
in establishing the pro-inflammatory phenotype upon earlier. Next, we investigated the expression patterns of
these 18 target genes in our microarray data. As
expected, all 18 genes showed the pro-inflammatory
phenotype (Figure 5) (p = 2.94 × 10-4). We also sampled
10 random sets of 18 genes each (generated using the
RSAT tool [17]) for the pro-inflammatory pattern of
expression in our microarray data (Additional file 2,
Table S2) and found either no or very few genes (0, 1 or
2 out of 18) showing the pattern (p = 0.897). This target
gene analysis clearly establishes our predictions as
potentially genuine, biologically meaningful TF-TFBS
interactions for the pro-inflammatory phenotype. Validation of TF-TFBS interactions controlling the pro-
inflammatory genes Target genes highlighted in Table 3 were checked to look for inflammatory
gene expression pattern that is two-fold up regulation in N+L phenotype (compared to N) and further two-fold down regulation in T+L
phenotype. The relative expression patterns of 18 target genes found in our microarray data are shown. targets in the literature. Neither of these genes showed
the antibacterial expression pattern in the Foster et al
microarray data. inflammatory and antibacterial classes respectively. Basically, Foster et al and we hypothesize that during
tolerance although pro-inflammatory genes are down
regulated, antibacterial genes need to be continually
expressed. The two classes in their study were shown
to have differences in chromatin modifications indicat-
ing transient silencing of some pro-inflammatory genes
while priming of antibacterial gene expression [6]. However, they did not attempt to identify any global
TF-TFBS interactions that can attest to transcriptional
signatures responsible for the distinction between the
two classes and hence to LPS tolerance. We have iden-
tified and characterized such distinctive interactions
employing pattern search algorithms combined with Validation of TF-TFBS interactions controlling the pro-
inflammatory genes A
similar analysis on the two motifs (and the correspond-
ing profile TFs, Maz and Foxj2, Table 2) from the origi-
nal 228 random genes used in the TFBS prediction
algorithms, identified a different set of 18 target genes
(Additional file 3, Table S3) none of which had the pro-
inflammatory expression pattern. As a further step to
verify the role of these validated TF-TFBS interactions
in establishing the pro-inflammatory phenotype upon When we checked Prrx2, the only TF profile that was
identified in our TFBS prediction from the antibacterial
genes, it detected two genes (Ifi204, Pdgfra) as potential Table 3 Biologically validated target genes of profile TFs predicted from the pro-inflammatory gene set
Matrix Accession
Number
TF
Target Genes
M01014
Sox5
Smad5, Smad1, Smad7, Sox6, Sox5, Mir125b2, Mir34a, Mir224, Mir15a, Mir125b1, Lipe, Sry
M01014
SRY
Slc9a3r2, Wt1, Akr1b10, Zfp748, Hdac3, Smad3, Ar, Importin beta, Ep300, Kpnb1, Kpna, Znf208, Sry, Amh, Ptgds
M00699
IRF-8
Spi1, Irf1, Trim21, Cops2, Irf2, Il12b, Il1b, Cybb, B2 m, Cbl, Irf4, Nfatc1, Ttraf6, Stat1, Etv6, beta2-mg, Cdkn2b, Il-12
p40, H-2Dd, H-2Kb, H-2Ld
M01014
Sox13
Smad7, Fgf3
M01014
Sox4
Mir199a1, Mir27b, Mir199a2, Mir206, Mir29c, Mir107, Mir34a, Mir95, Mir17, Mir199b, Mirn292, Mirn101b, Cebpa, Sdcbp,
Tcf4
M01014
Sox2
Pou5f1, Pou2f1, Pax6, Lbx1, Pdx1, Meis1, Asc, Golga6, Nkx2-3, Otp, Dlx5, Otx1, Dlx4, Isl1, Zfhx3, Fbxo15, Fgf4, Hrc,
Nanog, Spp1, Zscan10
M01014
Sox9
Ep300, Nr5a1, Kpnb1, Crebbp, Smad3, Smad2, Amh, Mia, Med12, Maf, Importin beta, Calmodulin, Ppargc1a,
Ncadherin, Col2a1
M01014
Sox15
Fhl3, Pou5f1
M00233
Mef-2
Smarca4, Hdac4, Hdac9, Hdac7, Hdac5, Nfat, Mapk14, Thra, Ep300, Ckm, Myog, Mef2 d, Jun, Slc2a4, Srf, Mck
M00062
IRF-1
Agtr2, C2ta, Nos2, H-2kb, Ptgs2, Tlr3, H2-Dd, IL-12, IL-7R, Stat1, Ciita, Nfkb1, Rela, Stat5, Tap1, Vcam1, Psmb9, Ifnb1,
Stat3, Irf8, Crebbp, Smarca4, Cybb
M01014
Sox6
Cenpk, Sox5, Dazap2, Hdac1, Ctnnb1, Pdx1, Ctbp2, Mir29a, Mir221, Mir222, Mir29c, Mir126, Ccnd1, Fgf3, Hbb-y
M01014
Sox18
Mef2c, Vcam1 ogically validated target genes of profile TFs predicted from the pro-inflammatory gene set
sion
TF
Target Genes Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Page 8 of 12 Figure 5 Expression patterns pro-inflammatory target gene. Target genes highlighted in Table 3 were checked to look for inflammatory
gene expression pattern that is two-fold up regulation in N+L phenotype (compared to N) and further two-fold down regulation in T+L
phenotype. The relative expression patterns of 18 target genes found in our microarray data are shown. Figure 5 Expression patterns pro-inflammatory target gene. Discussion Due to the
small size of this data set, subsequent analyses of motif
search, and target identification and validation for this
class of genes did not merit much attention. However,
genes such as Lcn2 [18] and Tirap [19] in this list
(Table S1) have shown antimicrobial activity. Interest-
ingly Lcn2 suppressed LPS-induced inflammatory cyto-
kines in macrophages [20] indicating an additional anti-
inflammatory role for this gene. It is likely that a num-
ber of antibacterial genes in our microarray data main-
tain the same level of expression at both the N+L and T
+L stages to provide persistent bacterial protection and
hence it would be worthwhile to analyze a larger set of
these genes by filtering with a less stringent condition. g
y
g
g
Based on the concept of coordinate gene expression
being controlled by same or similar TFs binding to their
cognate binding motifs, we identified respectively four
and one TFBS motifs in the upstream regulatory regions
of the pro-inflammatory and antibacterial genes (Figure
4). The essence of the specificity of these TF-TFBS
interactions was provided by the intersection of the pre-
dictions from three tools of differing algorithmic
approaches. Additionally, when compared to a random
gene background the prediction scores for the four pro-
inflammatory motifs were significantly higher in the test
data set (Tables 1 &2). Our predictions identified inter-
feron regulatory factors 1 and 8 (IRF-1 and IRF-8) as
pro-inflammatory TFs (Figure 4, Table 1) and these pro-
teins have been clearly implicated in macrophage-asso-
ciated innate immunity [21-23]. A limitation to this study is the lack of the tolerant
(T) stage in the Foster et al [6] data set unlike the
Mages et al [10] data. By employing the N+L stage
instead of T (Figures 1, 2 and 5) we are likely to miss
some genes (false negatives) that are tolerizable. How-
ever, the filtering approach that we employed is logically
sound and did result in a significant number (228) of
pro-inflammatory genes. Our motif prediction tools con-
verged on 4 TF binding motifs that could co-ordinately
regulate these genes (Figure 4). From a biological per-
spective, the 18 genes that are validated from the litera-
ture regards to being transcriptional targets of the four
predicted profile TFs are most likely to be genuine can-
didates for establishing and/or maintaining the pro-
inflammatory phenotype. Discussion As a basis for characterizing transcriptomic signatures
in LPS tolerance, we differentiated pro-inflammatory
and antibacterial gene expression in LPS-stimulated
macrophages. The genes in the Foster et al [6] micro-
array data were classified into pro-inflammatory and
antibacterial sets using biologically relevant filters
(Figures 1 and 2). Authors of that study had performed
similar categorization to identify “tolerizable” and
“non-tolerizable” classes corresponding to our pro- Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Page 9 of 12 IRF1, IRF2, Stat1, Stat3 and Nfkb1 [8] that are essential
members of presumptive transcriptional regulatory net-
works. Nfkb1 has been identified as a major player in
the downstream signalling pathways of LPS-stimulated
macrophages [24] with a crucial role in the transcrip-
tional regulation of a number of target genes [9,25]. More importantly, some of these target genes, such as
Trim21, Ptgs2 and Nos2 (Figure 5) that show the shar-
pest drop in gene expression upon prolonged LPS treat-
ment have been implicated in tolerance [26-28]. Nfkb1
acts as the controlling TF in LPS-induced expression of
Ptgs2 and Nos2 [29] while acting downstream of
Trim21 [26]. Here, since the microarray data represents
a static view of gene expression, we cannot determine
whether the Nfkb1 gene acts up or downstream of these
genes as they all (Nfkb1, Trim21, Ptgs2 and Nos2) show
the prototypical pro-inflammatory phenotype of down
regulation (Figure 5) in induction of tolerance. A caveat
to this classification may be the case of Nos2 that shows
up in the pro-inflammatory category in our analysis. This gene has a direct role in killing intracellular patho-
gens [30-32] even though it shows down regulation in
the tolerant state. A reason for the presence of such
genes in our classification maybe due to the non-linear-
ity of TF-TFBS interactions in that the dynamic modula-
tion of transcriptional target gene expression does not
always correlate with the corresponding TF binding to
its site. literature-based validation of the target genes in the
microarray data. Our filtering approach deducted only a small number
(18) of antibacterial genes since the cut-off of even one-
fold step-wise increase from the N to N+L to T+L
stages along with the p-value threshold seems to be
highly stringent and this may indicate a highly specific
antibacterial phenotype for these genes. Discussion We validated the TF-TFBS predictions in the pro-
inflammatory class of genes by manually identifying the
corresponding target genes implicated to be under con-
trol of the predicted TFs. This analysis produced a sig-
nificant number of target genes and a small proportion
of them (18 out of 141) showed the pro-inflammatory
specific pattern of gene expression (Table 3, Figure 5). A similar gene expression pattern of the 18 genes identi-
fied in our analysis with data from the Mages et al [10]
microarray analysis (Table S4) confirms the genuine
pro-inflammatory phenotype of these transcriptional tar-
gets in macrophage tolerance induction, and indicates
that the four predicted TFs most likely control the tran-
scriptional regulation of these genes to establish the
phenotype. A number of genes in this list have been
experimentally associated with LPS-mediated macro-
phage activation including some important TFs such as It is interesting to note that although the exhaustive
work of Ramsey et al [8] looked at the dynamics of
transcriptional programs in LPS-stimulated macro-
phages, they did not characterize the differential expres-
sion of the two categories of tolerant genes as belonging
to the pro-inflammatory and antibacterial classes. We
observed a number of common genes between our 228
pro-inflammatory set and their list of 1960 differentially
expressed genes probably indicating a mixture of both
classes of genes in their data. Litvak et al [9] have impli-
cated one such gene, Cebpd, a TF, in a regulatory circuit Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Page 10 of 12 Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Differentially expressed genes y
p
g
The Foster et al microarray dataset (GSE7348) was down-
loaded from the NCBI-GEO database [33]via FTP proto-
col. This data set was derived from murine (C57BL/6
strain) bone marrow macrophages left untreated (N), sti-
mulated with LPS for 24 hours and then re-stimulated for
4 hours (T+L) or treated only with the second stimulation
for 4 hours (N+L). RNAs from these 3 conditions were
hybridized to the Affymetrix Mouse Genome 430 2.0
arrays (in duplicates) to get raw gene expression values for
45101 probe sets [6]. Differentially expressed genes were
selected based on their expression level differences in var-
ious experimental conditions. For gene selection, in addi-
tion to fold change, we also performed Student’s two-
tailed t-test by assuming heteroscedasticity between the
naïve and tolerant macrophages to prove significant differ-
ences in expression between the two conditions. For a
pro-inflammatory gene, the ratio of average gene expres-
sion values in N+L to N and T+L to N+L should be > one
and < one respectively, and have the t-test p-value of <
0.05. On the other hand, for an antibacterial gene, the
ratio of average gene expression values in both N+L to N
and T+L to N+L should be > one, and have the t-test p-
value of <0.05. In this study, we utilized random mouse
genes as background and they were selected using the
RSAT tool [17]. The Mages et al [10] data was down-
loaded from GEO (GSE8621) and appropriate differential
gene expression filters (based on fold change) were applied
to compare with the pro-inflammatory transcriptional tar-
gets identified in our analysis of the Foster et al data. MEME It works by searching for repeated, ungapped sequence
patterns in the input DNA with statistical significance. The non-default parameters used for this analysis com-
prised of number of motifs (30), maximum width (15),
and mode of motif distribution (zero or one per
sequence, zoops). Additionally the revcomp parameter
was used to search for TFBS in both strands. MEME
identifies motifs without any concern to biological
validation. Sequence retrieval discriminating between transient and persistent TLR4-
stimulated signals. A search for the Cebpd binding
motif in the regulatory regions of our two classes of
genes (similar to their analysis) is likely to shed more
light on the varying gene expression patterns in LPS tol-
erance induction. Regulator modules are generally located in the upstream
and near the promoter regions of a gene. We considered
this as our basis to retrieve the 1000 to +300 region,
with respect to TSS, of all selected test and random
genes. Gene sequences were retrieved from NCBI using
a local tool [34] and repeats were masked using Repeat-
Masker [35]. Conclusions By combining distinct gene expression array data with
motif scanning and literature-based biological validation,
we have identified and characterized transcriptomic sig-
natures categorizing pro-inflammatory and antibacterial
classes of genes in LPS tolerant macrophages. We iden-
tified 228 pro-inflammatory and 18 antibacterial genes
likely to be transcriptionally regulated by four and one
TFs respectively. Further, employing literature resources,
we observed a number of target genes corresponding to
the predicted profile TFs in the pro-inflammatory set
and a subset of these targets clearly showed the pro-
inflammatory gene expression pattern corresponding to
LPS tolerance. Identification of TFBS We used three TFBS prediction tools viz., MEME [12],
MotifModeler [13], and PASTAA [14]. We performed
this analysis to both our test and random sets of genes. Description and parameters used for these tools are as
follows: MotifModeler Gene UIDs and corresponding expression values of a
co-regulated set of genes were given as input and this
software works by taking a set of random motifs of
fixed size and mapping them onto putative regulatory
regions of genes of interest. A linear model was estab-
lished by considering the expression values and the effi-
cacy of selected motifs. In this model each motif was
evaluated based on its contribution to transcriptional
regulation. This was iterated many times to calculate a
cumulative transcription contribution score (TCS) that
was used for motif selection. Least square method was
used to dictate inhibitory and stimulatory effects of the
predicted motifs. For the analysis of random gene data,
we used random expression values, generated by a PERL
script, within the range of the maximum and minimum
expression values of test genes. Additional material 18. Saiga H, Nishimura J, Kuwata H, Okuyama M, Matsumoto S, Sato S,
Matsumoto M, Akira S, Yoshikai Y, Honda K, Yamamoto M, Takeda K:
Lipocalin 2-Dependent Inhibition of Mycobacterial Growth in Alveolar
Epithelium. J Immunol 2008, 181(12):8521-8527. 18. Saiga H, Nishimura J, Kuwata H, Okuyama M, Matsumoto S, Sato S,
Matsumoto M, Akira S, Yoshikai Y, Honda K, Yamamoto M, Takeda K:
Lipocalin 2-Dependent Inhibition of Mycobacterial Growth in Alveolar
Epithelium. J Immunol 2008, 181(12):8521-8527. 19. Jeyaseelan S, Young SK, Yamamoto M, Arndt PG, Akira S, Kolls JK,
Worthen GS: Toll/IL-1R Domain-Containing Adaptor Protein (TIRAP) Is a
Critical Mediator of Antibacterial Defense in the Lung against Klebsiella
pneumoniae but Not Pseudomonas aeruginosa. J Immunol 2006,
177(1):538-547. Literature-based validation for target genes We collected a list of genes (from TRANSFAC and IPA)
that have been biologically identified as targets to the
profile TFs that were discovered in motif prediction. We
compared this list with the list of genes in the microar-
ray that passed through a two-fold filter for the pro-
inflammatory expression pattern. By normalizing the
highest expression value in the N condition of the raw
microarray data (45101 probe values) to 1, we calculated
relative expression values for the filtered target genes
corresponding to the N, N+L and T+L conditions (Fig-
ure 5). p
y
p
,
( )
9. Litvak V, Ramsey SA, Rust AG, Zak DE, Kennedy KA, Lampano AE, Nykter M,
Shmulevich I, Aderem A: Function of C/EBP[delta] in a regulatory circuit
that discriminates between transient and persistent TLR4-induced
signals. Nat Immunol 2009, 10(4):437-443. g
10. Mages J, Dietrich H, Lang R: A genome-wide analysis of LPS tolerance in
macrophages. Immunobiology 2007, 212(9-10):723-737. 11. MeV: MultiExperiment Viewer. [http://www.tm4.org/mev.html]. 10. Mages J, Dietrich H, Lang R: A genome-wide analysis of LPS tolerance in
macrophages. Immunobiology 2007, 212(9-10):723-737. macrophages. Immunobiology 2007, 212(9-10):723-737. 11. MeV: MultiExperiment Viewer. [http://www.tm4.org/mev.html]. 11. MeV: MultiExperiment Viewer. [http://www.tm4.org/mev.html]. 12. Bailey TL, Elkan C: Fitting a mixture model by expectation maximization
to discover motifs in biopolymers. Proc Int Conf Intell Syst Mol Biol 1994,
2:28-36. 12. Bailey TL, Elkan C: Fitting a mixture model by expectation maximization
to discover motifs in biopolymers. Proc Int Conf Intell Syst Mol Biol 1994,
2:28-36. 13. Liu Y, Taylor MW, Edenberg HJ: Model-based identification of cis-acting
elements from microarray data. Genomics 2006, 88(4):452-461. 13. Liu Y, Taylor MW, Edenberg HJ: Model-based identification of cis-acting
elements from microarray data. Genomics 2006, 88(4):452-461. y
14. Roider HG, Manke T, O’Keeffe S, Vingron M, Haas SA: PASTAA: identifying
transcription factors associated with sets of co-regulated genes. Bioinformatics 2009, 25:435-442. 14. Roider HG, Manke T, O’Keeffe S, Vingron M, Haas SA: PASTAA: identifying
transcription factors associated with sets of co-regulated genes. Bioinformatics 2009, 25:435-442. Motif selection and their corresponding profile TFs 3. Karin M, Lawrence T, Nizet V: Innate Immunity Gone Awry: Linking Microbial
Infections to Chronic Inflammation and Cancer 2006, 124(4):823-835. Top 70% of the predicted motifs were selected and com-
pared in all tools. MEME predicts the consensus
sequences of all possible hits in the promoter region of
a given set of genes but does not provide any annotated
information about the predictions. Hence, we took the
position weight matrices (PWM) of the predicted con-
sensus sequences and searched in TRANSFAC [15] for
the motif accession numbers using the TOMTOM tool
[36]. MotifModeler and PASTAA provide motif acces-
sion numbers for the predicted motifs. We used
TRANSFAC to manually map these motifs to TFs. 4. Liew FY, Xu D, Brint EK, O’Neill LAJ: Negative regulation of Toll-like
receptor-mediated immune responses. Nat Rev Immunol 2005,
5(6):446-458. 5. West M, Heagy W: Endotoxin tolerance: A review. Crit Care Med 2002, 30(1
Supp):S64-S73. 6. Foster SL, Hargreaves DC, Medzhitov R: Gene-specific control of
inflammation by TLR-induced chromatin modifications. Nature 2007,
447(7147):972-978. ,
g
,
p
inflammation by TLR-induced chromatin modifications. Nature 2007,
447(7147):972-978. 7. Roach JC, Smith KD, Strobe KL, Nissen SM, Haudenschild CD, Zhou D,
Vasicek TJ, Held GA, Stolovitzky GA, Hood LE, Aderem A: Transcription
factor expression in lipopolysaccharide-activated peripheral-blood-
derived mononuclear cells. Proceedings of the National Academy of Sciences
2007, 104(41):16245-16250. 8. Ramsey SA, Klemm SL, Zak DE, Kennedy KA, Thorsson V, Li B, Gilchrist M,
Gold ES, Johnson CD, Litvak V, Navarro G, Roach JC, Rosenberger CM,
Rust AG, Yudkovsky N, Aderem A, Shmulevich I: Uncovering a Macrophage
Transcriptional Program by Integrating Evidence from Motif Scanning Authors’ contributions 20. Zhang J, Wu Y, Zhang Y, LeRoith D, Bernlohr DA, Chen X: The Role of
Lipocalin 2 in the Regulation of Inflammation in Adipocytes and
Macrophages. Mol Endocrinol 2008, 22(6):1416-1426. 20. Zhang J, Wu Y, Zhang Y, LeRoith D, Bernlohr DA, Chen X: The Role of
Lipocalin 2 in the Regulation of Inflammation in Adipocytes and
Macrophages. Mol Endocrinol 2008, 22(6):1416-1426. RKK participated in the conception of the study, designed and carried out all
the experiments, analyzed the data and helped draft the manuscript. NBP
conceived of the study, participated in its design and coordination and
drafted the manuscript. Both authors read and approved the final
manuscript. 21. Dror N, Alter-Koltunoff M, Azriel A, Amariglio N, Jacob-Hirsch J, Zeligson S,
Morgenstern A, Tamura T, Hauser H, Rechavi G, Ozato K, Levi B-Z:
Identification of IRF-8 and IRF-1 target genes in activated macrophages. Mol Immunol 2007, 44(4):338-346. Competing interests 22. Negishi H, Fujita Y, Yanai H, Sakaguchi S, Ouyang X, Shinohara M,
Takayanagi H, Ohba Y, Taniguchi T, Honda K: Evidence for licensing of
IFN-gamma-induced IFN regulatory factor 1 transcription factor by
MyD88 in Toll-like receptor-dependent gene induction program. Proceedings of the National Academy of Sciences 2006, 103(41):15136-15141. The authors declare that they have no competing interests. Received: 7 September 2010 Accepted: 21 September 2010
Published: 21 September 2010 Received: 7 September 2010 Accepted: 21 September 2010
Published: 21 September 2010 23. Vila-del Sol V, Punzon C, Fresno M: IFN-{gamma}-Induced TNF-{alpha}
Expression Is Regulated by Interferon Regulatory Factors 1 and 8 in
Mouse Macrophages. J Immunol 2008, 181(7):4461-4470. p
PASTAA This software takes ENSEMBL gene ids as input and
ranks them based on the annotated binding affinities of
549 vertebrate TRANSFAC TFs to their promoter
regions. Simultaneously, it also ranks them based on
their tissue specificities derived from expression data. These two lists are compared for high ranked common
genes and an iterated hypergeometric test is performed
to deduce the relationships between TFs and the list of
genes and corresponding p-values calculated. Lower the
p-value better the prediction. Page 11 of 12 Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Acknowledgements 15. Matys V, Fricke E, Geffers R, Gossling E, Haubrock M, Hehl R, Hornischer K,
Karas D, Kel AE, Kel-Margoulis OV, Land S, Lewicki-Potapov B, Michael H,
Münch R, Reuter I, Rotert S, Saxel H, Scheer M, Thiele S, Wingender E:
TRANSFAC(R): transcriptional regulation, from patterns to profiles. Nucl
Acids Res 2003, 31(1):374-378. 15. Matys V, Fricke E, Geffers R, Gossling E, Haubrock M, Hehl R, Hornischer K,
Karas D, Kel AE, Kel-Margoulis OV, Land S, Lewicki-Potapov B, Michael H,
Münch R, Reuter I, Rotert S, Saxel H, Scheer M, Thiele S, Wingender E:
TRANSFAC(R): transcriptional regulation, from patterns to profiles. Nucl
Acids Res 2003, 31(1):374-378. We thank M. H. Kaplan for critical review of the manu-
script. RKK was supported by a research fellowship
from the IUPUI Graduate Office. 16. Ingenuity Systems. [http://www.ingenuity.com]. 17. Thomas-Chollier M, Sand O, Turatsinze J-V, Janky Rs, Defrance M, Vervisch E,
Brohee S, van Helden J: RSAT: regulatory sequence analysis tools. Nucl
Acids Res 2008, 36(suppl_2):W119-127. Additional material
Additional file 1: Table S1. Additional file 2: Table S2. Additional file 3: Table S3. Additional file 4: Table S4. 17. Thomas-Chollier M, Sand O, Turatsinze J-V, Janky Rs, Defrance M, Vervisch E
Brohee S, van Helden J: RSAT: regulatory sequence analysis tools. Nucl
Acids Res 2008, 36(suppl_2):W119-127. References 24. Dos Santos S, Delattre A.-I, De Longueville F, Bult H, Raes M: Gene
Expression Profiling of LPS-Stimulated Murine Macrophages and Role of
the NF-kB and PI3K/mTOR Signaling Pathways. Annals of the New York Page 12 of 12 Page 12 of 12 Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 Academy of Sciences 2007, 1096(Signal Transduction Pathways Part D
Inflammatory Signaling Pathways and Neuropathology) 70-77. 25. Kollet JI, Petro TM: IRF-1 and NF-[kappa]B p50/cRel bind to distinct
regions of the proximal murine IL-12 p35 promoter during costimulation
with IFN-[gamma] and LPS. Molecular Immunology 2006, 43(6):623-633. 26. Yoshimi R, Chang T-H, Wang H, Atsumi T, Morse HC, Ozato K: Gene
Disruption Study Reveals a Nonredundant Role for TRIM21/Ro52 in NF-
{kappa}B-Dependent Cytokine Expression in Fibroblasts. J Immunol 2009,
182(12):7527-7538. 27. Cao C, Matsumura K, Yamagata K, Watanabe Y: Involvement of
cyclooxygenase-2 in LPS-induced fever and regulation of its mRNA by
LPS in the rat brain. Am J Physiol Regul Integr Comp Physiol 1997, 272(6):
R1712-1725. 28. Dias MB, Almeida MC, Carnio EC, Branco LGS: Role of nitric oxide in
tolerance to lipopolysaccharide in mice. J Appl Physiol 2005,
98(4):1322-1327. 29. Spitzer JA, Zheng M, Kolls JK, Vande Stouwe C, Spitzer JJ: Ethanol and LPS
modulate NF-kappaB activation, inducible NO synthase and COX-2 gene
expression in rat liver cells in vivo. Front Biosci 2002, 7:a99-108. 30. Utaisincharoen P, Anuntagool N, Arjcharoen S, Limposuwan K, Chaisuriya P,
Sirisinha S: Induction of iNOS expression and antimicrobial activity by
interferon-beta is distinct from IFN-gamma in Burkholderia
pseudomalleiinfected mouse macrophages. Clinical & Experimental
Immunology 2004, 136(2):277-283. 31. Tarasenko O, Scott A, Soderberg L, Ponnappan U, Alusta P: Killing of
Bacillus spores is mediated by nitric oxide and nitric oxide synthase
during glycoconjugate-enhanced phagocytosis. Glycoconjugate Journal
2010, 27(1):13-25. 32. Hazlett LD, McClellan S, Goshgarian C, Huang X, Thakur A, Barrett R: The
Role of Nitric Oxide in Resistance to P. aeruginosa Ocular Infection. Ocular Immunology and Inflammation 2005, 13(4):279-288. 33. Barrett T, Troup DB, Wilhite SE, Ledoux P, Rudnev D, Evangelista C, Kim IF,
Soboleva A, Tomashevsky M, Edgar R: NCBI GEO: mining tens of millions
of expression profiles–database and tools update. Nucl Acids Res 2007,
35(suppl_1):D760-765. 34. Grobe M: Get NCBI sequences for genes or specified regions. [http://
discern.uits.iu.edu:8421/view-sequences.html]. 34. Grobe M: Get NCBI sequences for genes or specified regions. [http://
discern.uits.iu.edu:8421/view-sequences.html]. 35. Smit AFA, Hubley R, Green P: RepeatMasker Open-3.0. 1996-2010. [http://
repeatmasker.org]. 35. Smit AFA, Hubley R, Green P: RepeatMasker Open-3.0. 1996-2010. [http://
repeatmasker.org]. 36. Gupta S, Stamatoyannopoulos JA, Bailey TL, Noble WS: Quantifying
similarity between motifs. Genome Biol 2007, 8(2):R24. 37. Beissbarth T, Speed TP: GOstat: find statistically overrepresented Gene
Ontologies within a group of genes. Bioinformatics 2004, 20(9):1464-1465. Kollipara and Perumal Immunome Research 2010, 6:5
http://www.immunome-research.com/content/6/1/5 doi:10.1186/1745-7580-6-5
Cite this article as: Kollipara and Perumal: Motif prediction to distinguish
LPS-stimulated pro-inflammatory vs. antibacterial macrophage genes. Immunome Research 2010 6:5. Submit your next manuscript to BioMed Central
and take full advantage of:
• Convenient online submission
• Thorough peer review
• No space constraints or color figure charges
• Immediate publication on acceptance
• Inclusion in PubMed, CAS, Scopus and Google Scholar
• Research which is freely available for redistribution
Submit your manuscript at
www.biomedcentral.com/submit Submit your next manuscript to BioMed Central
and take full advantage of: Submit your next manuscript to BioMed Central
and take full advantage of: • Convenient online submission
|
https://openalex.org/W4307553650
|
https://www.frontiersin.org/articles/10.3389/fnsys.2022.855107/pdf
|
English
| null |
Effects of exercise on the sleep microarchitecture in the aging brain: A study on a sedentary sample
|
Frontiers in systems neuroscience
| 2,022
|
cc-by
| 7,275
|
TYPE Brief Research Report
PUBLISHED 26 October 2022
DOI 10.3389/fnsys.2022.855107 TYPE Brief Research Report
PUBLISHED 26 October 2022
DOI 10.3389/fnsys.2022.855107 Effects of exercise on the sleep
microarchitecture in the aging
brain: A study on a sedentary
sample OPEN ACCESS
EDITED BY
Axel Hutt,
Inria Nancy - Grand-Est Research
Centre, France
REVIEWED BY
Michelangelo Maestri,
University of Pisa, Italy
Oleg Lyamin,
UCLA Department of Psychiatry,
United States
*CORRESPONDENCE
Paul S. García
pg2618@cumc.columbia.edu
RECEIVED 14 January 2022
ACCEPTED 26 August 2022
PUBLISHED 26 October 2022
CITATION
Cassim TZ, McGregor KM, Nocera JR,
García VV, Sinon CG, Kreuzer M and
García PS (2022) Effects of exercise on
the sleep microarchitecture
in the aging brain: A study on
a sedentary sample. Front. Syst. Neurosci. 16:855107. doi: 10.3389/fnsys.2022.855107 OPEN ACCESS
EDITED BY
Axel Hutt,
Inria Nancy - Grand-Est Research
Centre, France
REVIEWED BY
Michelangelo Maestri,
University of Pisa, Italy
Oleg Lyamin,
UCLA Department of Psychiatry,
United States
*CORRESPONDENCE
Paul S. García
pg2618@cumc.columbia.edu Tuan Z. Cassim1, Keith M. McGregor2,3, Joe R. Nocera4,5,6,
Violet V. García7, Christopher G. Sinon8, Matthias Kreuzer9
and Paul S. García1* Tuan Z. Cassim1, Keith M. McGregor2,3, Joe R. Nocera4,5,6,
Violet V. García7, Christopher G. Sinon8, Matthias Kreuzer9
and Paul S. García1* Tuan Z. Cassim1, Keith M. McGregor2,3, Joe R. Nocera4,5,6,
Violet V. García7, Christopher G. Sinon8, Matthias Kreuzer9
and Paul S. García1* 1Department of Anesthesiology, Neuroanesthesia Division, Columbia University Medical Center,
New York-Presbyterian Hospital, New York, NY, United States, 2Department of Clinical and
Diagnostic Sciences, School of Health Profession, University of Alabama at Birmingham,
Birmingham, AL, United States, 3Birmingham Veterans Affairs Geriatric Research, Education, and
Clinical Center (GRECC), Birmingham, AL, United States, 4VA Rehabilitation R&D Center for Visual
and Neurocognitive Rehabilitation, Atlanta VAMC, Decatur, GA, United States, 5Department of
Neurology, Emory University School of Medicine, Atlanta, GA, United States, 6Department of
Rehabilitation Medicine, Emory University School of Medicine, Atlanta, GA, United States, 7Stamps
President’s Scholars Program, Georgia Institute of Technology, Atlanta, GA, United States, 8Yerkes
National Primate Research Center, Neuropharmacology and Neurologic Diseases, Emory
University, Atlanta, GA, United States, 9Department of Anesthesiology, School of Medicine,
Technical University of Munich, Munich, Germany Cassim TZ, McGregor KM, Nocera JR,
García VV, Sinon CG, Kreuzer M and
García PS (2022) Effects of exercise on
the sleep microarchitecture
in the aging brain: A study on
a sedentary sample. Front. Syst. Neurosci. 16:855107. doi: 10.3389/fnsys.2022.855107 Having a healthy sleep pattern plays a vital role in one’s overall health. Sleep in
the elderly is characterized by decreased slow-wave sleep and an increase of
REM sleep. Furthermore, quantitative electroencephalographic (qEEG) studies
have shown an age-related attenuation of total EEG power in sleep. aging, aerobic exercise, physical activity, sleep, deep sleep (slow wave sleep, SWS),
electroencephaloagraphy (EEG) Frontiers in Systems Neuroscience Introduction Neurophysiological recordings serve as reliable
biomarkers of sleep stages even when recorded via portable
at-home sleep devices (Levendowski et al., 2017). Lastly, advanced age also affects the inhibitory signaling
of cortical networks (Lariviere et al., 2007). The efficacy of
these inhibitory cortical networks strongly correlates with sleep
quality (Paul-Savoie et al., 2012). Exercise can reverse the loss
of inhibitory signaling among cortical networks to improve
neurologic performance (McGregor et al., 2017). Hence, an
increased level of physical fitness can lead to improved sleep
quality (King et al., 2008) in older adults that were previously
sedentary. While there are numerous reports of the human and animal
studies of potential neuroprotective effect of acute exercise
through improved sleep (Kredlow et al., 2015; Panagiotou
et al., 2018), the detailed changes of aerobic exercise have
on sleep microarchitecture (i.e., quantitative EEG features)
is not well studied. Further, we know of no studies that
have investigated quantitative EEG in older adults after a
longer-term physical activity intervention. In our study, we
demonstrate the feasibility of an “in-home” device for capturing
neurophysiologic data during natural sleep in older adults while
we also examined the effects of aerobic exercise on the sleep
microarchitecture we decode changes in the spectral features
in this aged sample. Given the evidence from the literature, we
expect to see an improvement in SWS associated with aerobic
exercise. With aging, sleep processes can be disrupted causing
unhealthy sleep patterns and sometimes influence pathogenesis
of sleep disorders (Zhong et al., 2019). In older adults,
complaints about sleep include: light sleep, spontaneous waking
during the night, daytime sleepiness, and a decrease in
overall sleep time (Bliwise et al., 1992; Foley et al., 1995;
Vitiello et al., 2004). Sleep disorders are also associated
with cognitive decline (Dzierzewski et al., 2018), yet they
remain a commonly underdiagnosed problem. In the literature,
slow wave sleep (SWS) also known as deep sleep has
shown to be a crucial biomarker that is associated with
restorative properties such as memory consolidation and
improvements in age-related memory integrity (Papalambros
et al., 2017; Berres and Erdfelder, 2021). However, one of
the most prevalent sleep problems observed in the aged
population is an increase in light sleep (N1 and N2) and
a decrease in SWS (Scullin, 2013; Scullin and Bliwise,
2015). Introduction positive implication in the clinical setting and in one’s overall
health. Exercise has shown to have beneficial effects to mitigate
age-related health problems. Exercise has also shown to have
neuroprotective qualities and help the cognitively vulnerable
populations and attenuate cognitive problems (Heyn et al., 2004;
Voss et al., 2013). The beneficial effects of exercise on sleep are
also well documented in the literature (Kredlow et al., 2015). Aerobic activity in particular has shown to have promising
results in promoting healthy sleep patterns and playing a
positive role in brain health (Hillman et al., 2008; Kaliman
et al., 2011; Hotting and Roder, 2013). Animals studies have
also shown positive effects of aerobic exercise has on sleep in
aged mice (Panagiotou et al., 2018). Thus, as an alternate to
pharmacological interventions for sleep disturbances exercise
has been proposed as a non-pharmacological intervention tool
to mitigate sleep problems (Chennaoui et al., 2015). Sleep, regulated by circadian and homoeostatic processes
plays a vital role in one’s overall health (Borbely et al., 2016). The sleep architecture in humans consists of intermittent
cycling among four sleep “stages” which are distinguished from
one another via neurophysiological recording of muscle and
brain activity. The ratios of these stages on a given night
of sleep for an individual vary based on many factors such
as age, sex, ethnicity, smoking habits, alcohol consumption,
etc. (Sahlin et al., 2009). Studies categorize sleep architecture
into vigilance stages also known as the macro-architecture
and spectral analysis of the electroencephalographic (EEG)
also known as the micro-architecture to understand variables
associated with sleep. Ocular movements distinguish rapid
eye movement (REM) EREM (rapid eye movement) sleep
from the three types of non-REM sleep: non-REM stage 1,
N1; non-REM stage 2, N2; non-REM stage 3, N3. Unique
EEG features pertaining to these stages and their analyses
are well documented in sleep literature (Silber et al., 2007). High frequency, low amplitude neurophysiological activity in
scalp electrodes over the frontal cortex are indicative of N1
and REM—so called “light” stages of sleep, because arousal is
common with mild stimuli (e.g., auditory stimulation at normal
volume). Stimuli of higher intensity (e.g., loud voice, tactile
stimulation) are necessary to arouse an individual from N2 to
N3 sleep, which are characterized by higher amplitude lower
frequency synchronized oscillations over frontal electrodes
(Schulz, 2008). Effects of exercise on the sleep
microarchitecture in the aging
brain: A study on a sedentary
sample However,
exercise has been shown to improve sleep across all age groups. In this
study, we used the Sleep ProfilerTM EEG Sleep Monitor to observe EEG
changes occurring during sleep following an aerobic exercise intervention. This study was done on older adults (N = 18, with only five subjects containing
both pre- and post-data of sufficient quality for analysis) with an age range
60–85 years. The aerobics regimen was performed three times weekly for
12-weeks commencing with 20-min sessions. The time of each session
progressed by 1–2 min/session as needed to a maximum time of 45 min
per session. The macro-architecture (sleep stages) and microarchitecture
(EEG) results were analyzed using MATLAB. For the microarchitecture, our
results showed more deep sleep following the aerobic exercise regimen. Furthermore, for the microarchitecture, out results shows an increase in total
EEG power post-exercise in both light (N1 and L1) and deep sleep (N2 and
N3). These preliminary changes in sleep the microarchitecture suggest that
non-pharmacologic methods might mitigate age-related EEG changes with
potential implications for neurocognitive health. COPYRIGHT
© 2022 Cassim, McGregor, Nocera,
García, Sinon, Kreuzer and García. This
is an open-access article distributed
under the terms of the Creative
Commons Attribution License (CC BY). The use, distribution or reproduction in
other forums is permitted, provided
the original author(s) and the copyright
owner(s) are credited and that the
original publication in this journal is
cited, in accordance with accepted
academic practice. No use, distribution
or reproduction is permitted which
does not comply with these terms. 01 Frontiers in Systems Neuroscience frontiersin.org Cassim et al. 10.3389/fnsys.2022.855107 Introduction Quantitative EEG (qEEG) studies have furthered the
understanding of age-related changes in sleep by decoding
specific spectral feature changes in the sleep EEG (Carrier
et al., 2001). Non-pharmacological methods to improve SWS
in this population is of special interest and can have Experimental design Approval for this study was granted by the local Institutional
Review Board and written informed consent was obtained from
each participant in the study in accordance with the Declaration
of Helsinki. This study was a sub-study of a larger study
(McGregor et al., 2017; Nocera et al., 2017) comparing the
effects of 12-weeks of aerobic exercise (bicycle “spin” class, three 02 Frontiers in Systems Neuroscience Frontiers in Systems Neuroscience frontiersin.org Cassim et al. 10.3389/fnsys.2022.855107 10.3389/fnsys.2022.855107 10.3389/fnsys.2022.855107 times each week) vs. 12-weeks of non-aerobic stretching and
balance “control” condition on motor responsiveness. A total of
28 separate participants between the ages of 60–89 took a Sleep
ProfilerTM EEG sleep monitor (a battery-powered, wireless, and
home-health sleep profiling device) home at least once for
collecting sleep data. Not all participants were able to reliably
provide data for the study. For our analysis we included a total
of n = 18 with baseline (pre-aerobic intervention) EEGs. To
investigate the effect of exercise on sleep we had n = 5 EEG
complete sets of both pre- and post-aerobic exercise recordings
of sufficient quality for analysis of microarchitecture. Due to
acquisition limitations on the device, stable EEG data was
collected from 18 out of 28 participants and of these 18 only 5
participants were able to provide pre/post intervention data. For the spin group, exercise intensity began at low levels
(target: 50% of HRR) and increased by 5% every week (as
deemed appropriate by the instructor) to a target maximum
of 80–85% HRR. Participants wishing to exceed this capacity
could do so for limited exercise intervals. Relative physical
exertion was assessed with the “talk-test” and a rating of
relative physical exertion estimation using the Borg 6–20
difficulty scale (6 = lowest effort; 20 = maximum effort)
during the sessions. For the spin group the instructor guided the participants
through a light 5-min warm up, then a workout phase that
included steady up-tempo cadences, sprints (increased rpm),
and climbs (increased resistance). As such, the spin exercise
routine employed an interval-based training approach. During
the workout phase, the target HRR was maintained by averaging
increases and decreases in intensity/HR. The goal was to
maintain within a 10% offset from the HRR goal during the
workout phase. Subjects Our participants (aged 60–85 years) were sedentary [defined
as engaging in less than 45 min of weekly moderate to vigorous
physical activity as assessed by the International Physical
Activity Questionnaire (IPAQ)], without major psychiatric
or neurological disease, -right-handed, and native English
speakers. Participants also required physician’s approval for
study participation. No participants had been hospitalized
within the previous 6 months or exhibited significant cognitive
impairment defined as < 23 on the Montreal Cognitive
Assessment (MoCA) (Nasreddine et al., 2005) and < 15 errors
on the American National Adult Reading Test (Grober and
Sliwinski, 1991). All interventions took place in a climate-controlled fitness
facility. Interventions occurred in the mornings (8–10 a.m. Eastern Time Zone) and ran on a rolling basis throughout
the calendar year. Sleep profiler and
electroencephalographic recording We
used
the
Sleep
ProfilerTM
EEG
Sleep
Monitor
(Advanced Brain Monitoring, Carlsbad, CA, United States),
a battery-powered, wireless, and home-health sleep profiling
device. Technical details regarding the device, sampling rates,
processing, and filtering can be accessed in the company
website.1 This device has been validated for staging of sleep and
is FDA cleared for aiding in the diagnosis of sleep disorders
(Levendowski et al., 2017). Participants took the device home
after being instructed on its use. Participants were asked
to clean their forehead using soap and water prior to the
placement of the electrodes to minimize impedance. If the
impedance level was too high a voice message alerted the
participants to adjust the sensors. The frontal EEG data were
recorded as a voltage difference between the two frontopolar
electrode positions AF7–AF8, according to the international
10–20 system (sampling rate = 256 Hz). The system also
recorded electrooculographic and electromyographic activity. The record of each subject was uploaded onto the Sleep
Profiler PortalTM, an internet-based software application. Each record was processed using an automatic scoring Experimental design For the control group, we monitored heart rate,
but did not specify a target HRR threshold for the balance
(progressive board/mat balance), stretching and light strength
training (using resistance bands) exercises employed. 1
https://cportal.b-alert.com/sleep-profiler/documentshelp Heart Rate Reserve = Max Heart Rate −Resting Heart Rate
(2) Sleep stages To investigate a possible impact of exercise on the spectral
information of the EEG, we calculated the power spectral density
(PSD) and the density spectral arrays (DSA) with the MATLAB
pwelch function with a frequency resolution of 1 Hz for the
30 s EEG segments the auto-profiler used to generate the sleep
scores. In order to minimize the impact of segments that may
contain a transition from one state to the other, we dismissed
these epochs from quantitative EEG analysis. Figure 2A presents the relative fraction of the different
sleep
stages
for
these
18
participants. The
distribution
of
stages
was
significantly
different
(p
<
0.001),
with
significantly higher proportions of N2, when compared to
N1 (corrected p = 0.038), to N3 (p < 0.001), and to L2
(p = 0.001). The participants spent 28% (15, 37%) in light
sleep, 14% (10, 21%) in REM sleep, and 40% (32, 46%) in
deep sleep (Figure 2B). This distribution was significantly
different (p < 0.001) with significantly more deep sleep
than REM (p < 0.001). Figure 2C compares the vigilance
state recording time percentage between baseline and after
aerobic exercise for the five participants in the subgroup
analysis. We did not find any significant differences in
the distribution of the data, but four out of these five
participants spent more time in deep sleep after the intervention
[AUC = 0.8 (0.4, 1)]. There was no difference in light
sleep [AUC = 0.6 (0.2, 1)] (Figure 2D). Further, four out
of five participants spent less time in REM sleep after the Sleep macro-architecture Figure 1 displays an exemplary hypnogram (1A) and
corresponding spectral array (1B) for a typical participant in the
sample. The corresponding hypnogram including the classical
sleep stages is presented as Supplementary Figure 1. The frontal
EEG data from the first to last sleep score for all (n = 18)
participants included in the study yielded approximately 635
(480, 898) min [median (1st, 3rd quartile)] of sleep for each of
the participants. Spectral electroencephalographic features In order to evaluate significant or relevant changes from
the PSD analyses which are presented in Figures 3, 4,
we only considered differences that occur in at least two
neighboring frequencies (2 Hz window), an approach that
was used earlier (Kreuzer et al., 2020). For AUC calculation
with 10k-fold bootstrapped 95% confidence intervals, we used
the MATLAB-based MES toolbox. We used this approach
to evaluate possible differences within groups between pre-
and post-exercise conditions as well as to find differences in
the EEG recorded post-exercise between the two intervention
groups. Statistical analysis We performed all statistical analyses using MATLAB. We
held the significance threshold at alpha = 0.05. Exercise regimen Consistent with our previous work (Nocera et al., 2017;
McGregor et al., 2018), the spin and control groups engaged in
exercise interventions beginning with 20 min of activity three
times a week for 12 weeks and led by a qualified instructor. In
both groups, the time of each session progressed based on the
recommendation of the instructor by 1–2 min to a maximum
time of 45 min per session at the end of the 12 weeks. For all
participants, we calculated their estimated maximum heart rate
using the Karvonen method (Eq. 1), and then calculated heart
rate reserve (HRR) as shown in Eq. 2. We used HRR values
to evaluate physical workload of participants throughout the
12-week interventions. Max Heart Rate = 220 bpm −Age
(1) (1) Max Heart Rate = 220 bpm −Age Heart Rate Reserve = Max Heart Rate −Resting Heart Rate
(2) Frontiers in Systems Neuroscience Frontiers in Systems Neuroscience 03 frontiersin.org 10.3389/fnsys.2022.855107 Cassim et al. 10.3389/fnsys.2022.855107 0.5, but the AUC was higher 0.7, we considered the effect
to be clinically relevant (Mandrekar, 2010). Furthermore,
to evaluate the distribution of the vigilance states, we
calculated the Friedman test with post-hoc Tukey-Kramer
correction. It has to be noted that for the analyses of the
matched pre-pot data the results from the statistical analysis
only carry little information because of the small sample
size. algorithm consistent with published guidelines American
Academy
of
Sleep
Medicine. Consistent
with
previous
literature (Levendowski et al., 2017), adjustments for age-
appropriate ratios of sleep stages were made. As is common
in evaluating patients for sleep apnea, the sleep profiler sub-
categorizes N2 into two separate stages (L2 and N2). Sleep
epochs were scored as L2 if markers indicative of a lighter
stage of sleep (e.g., elevated EMG activity or brief cortical
arousals) were also present during N2 (Levendowski et al.,
2017). The EEG recordings were stored in the edf format
for further analysis in MATLAB (MathWorks, Natick, MA,
United States). Sleep macro-architecture Based on the device-derived sleep stages from Sleep
ProfilerTM, we assessed the sleep macro-architecture (i.e., sleep
stages) during the baseline recordings in our 18 participants. For the five participants included in the exercise group with
valid pre- and post-intervention recordings, we also calculated
the change in the recording time percentage, i.e., the pre/post
change in percentage of sleep stages. Because we were especially
interested in the amount of restorative sleep (deep sleep
stages) rather than ratios of each individual stage of sleep, we
combined the light sleep stages (N1 and L2) and deep sleep
stages (N2 and N3). Frontiers in Systems Neuroscience Sleep stages Due to a limited sample size we chose the area under the
receiver operating curve (AUC) with 10k-fold bootstrapped
95% confidence intervals (CI) of the relative change in the
parameter from pre- to post-conditions as a measure of
effect size. If the 95% CI excluded 0.5, we considered the
effect statistically significant and if the 95% CI included 04 frontiersin.org Cassim et al. 10.3389/fnsys.2022.855107 FIGURE 1
Exemplary hypnogram (Top) and density spectral array (Bottom) of the pre-aerobics intervention EEG recording session. The hypnogram on top
shows the different vigilance states the subject transitioned throughout the night. These sleep stages were derived from the spectral information
provided in the density spectral array. The auto staging details of the sleep profiler are described in Levendowski et al. (2017). For example, light
sleep was indicated if there was strong activity in the EEG alpha-band around 10 Hz as indicated by red colors in this frequency range. FIGURE 1 Exemplary hypnogram (Top) and density spectral array (Bottom) of the pre-aerobics intervention EEG recording session. The hypnogram on top
shows the different vigilance states the subject transitioned throughout the night. These sleep stages were derived from the spectral information
provided in the density spectral array. The auto staging details of the sleep profiler are described in Levendowski et al. (2017). For example, light
sleep was indicated if there was strong activity in the EEG alpha-band around 10 Hz as indicated by red colors in this frequency range. Discussion intervention [AUC = 0.8 (0.4, 1)], Figure 2D. It appears that
exercise resulted in more deep sleep (and less REM) in this
small subgroup with pre- and post- intervention EEG data. Supplementary Table 1 contains the results from the statistical
tests. Our study confirmed results from previous literature
that showed an association between physical activity and an
improvement in sleep quality in older subjects. Specifically, our
aerobic exercise intervention resulted in increased restorative
sleep and an associated decrease of REM sleep in our
participants. This novel finding regarding vigilance states,
however, should still be considered somewhat preliminary
given our modest sample size. Additionally, the change in
sleep macro-architecture corresponded with changes in sleep
microarchitecture. Namely, the exercise intervention led to an
increase in overall EEG power in non-REM sleep. This is the first
time an FDA-approved take-home sleep device has been used to
examine the details of sleep neurophysiology in aerobic exercise
interventions in a vulnerable, aged population. Frontiers in Systems Neuroscience Sleep micro-architecture *Represent the TCassim. FIGURE 3
Power spectral density (PSD) for the different NREM sleep states recorded at baseline (n = 18). (A) For different levels of NREM sleep, i.e., N1, L2,
N2, and N3. (B) For light sleep and deep sleep. During deep sleep the power in the low (delta) frequencies was significantly higher and the
power in the high (beta) frequencies was significantly lower. that these EEG changes could indicate a potential therapeutic
benefit of aerobic exercise on cognition and a potential non-
pharmacologic approach could mitigate age-related attenuation
Other
researchers
have
demonstrated
that
1
h
of
vigorous
exercise
can
increase
slow-wave
sleep
and
decrease wake transitions in a small study consisting of FIGURE 2
Proportion of the vigilance states derived from the baseline recordings (n = 18) and the ratio of the vigilance states between the baseline and
post-aerobic exercise recordings from five participants. (A) Distribution of vigilance states for all 18 baseline recordings. (B) Distribution of light
sleep (N1 + L2), deep sleep (N2 + N3) and REM sleep at baseline (n = 18). (C) Relative change of the vigilance states from baseline to
post-aerobic exercise recordings (n = 5). (D) Relative change of light sleep, deep sleep and REM sleep from baseline to post-aerobic exercise
recordings (n = 5). *Represent the TCassim. Proportion of the vigilance states derived from the baseline recordings (n = 18) and the ratio of the vigilance states between the baseline and
post-aerobic exercise recordings from five participants. (A) Distribution of vigilance states for all 18 baseline recordings. (B) Distribution of light
sleep (N1 + L2), deep sleep (N2 + N3) and REM sleep at baseline (n = 18). (C) Relative change of the vigilance states from baseline to
post-aerobic exercise recordings (n = 5). (D) Relative change of light sleep, deep sleep and REM sleep from baseline to post-aerobic exercise
recordings (n = 5). *Represent the TCassim. FIGURE 3
Power spectral density (PSD) for the different NREM sleep states recorded at baseline (n = 18). (A) For different levels of NREM sleep, i.e., N1, L2,
N2, and N3. (B) For light sleep and deep sleep. During deep sleep the power in the low (delta) frequencies was significantly higher and the
power in the high (beta) frequencies was significantly lower. FIGURE 3
Power spectral density (PSD) for the different NREM sleep states recorded at baseline (n = 18). Sleep micro-architecture Figure 3A shows the PSD plots representing the power
in each EEG frequency for the different NREM sleep stages
and Figure 3B presents the PSD for light sleep and deep
sleep. As expected, for these 18 participants (pre-aerobic
intervention) the lighter stages of sleep (N1 + L2) have a
more uniform distribution of EEG frequency, characterized
by lower amplitude at the lower frequencies (< 5 Hz) and
higher amplitude at the higher frequencies (> 14 Hz) as
compared to deep sleep (N2 + N3). The horizontal gray
lines at the top of the figure depict differences in EEG
frequency that are determined to be statistically significant
(see Methods). In Figure 4 we compare the spectral EEG
features of deep sleep before and after the aerobic intervention
in the five participants in our intervention group. Changes
in the moderate and high frequencies predominate for deep
sleep as depicted in Figure 4A. For the five participants
with the matched EEG data, we observed a total EEG power
increase during in all five participants for non-REM sleep
(Figure 4B). Our findings are preliminary but provide some insight
into the complex relationship between aging, exercise, sleep,
and cognition. Total EEG power while sleeping, is known to
be lower in older individuals (Carrier et al., 2001). Similarly,
aged individuals and those with neurocognitive impairment
also exhibit a decline in overall EEG power (Kreuzer et al.,
2020). This might be related to an increased susceptibility
to disturbances of attention and arousal, especially in clinical
situations where anesthesia or sedation is used (e.g., post-
surgery). Our results confirm EEG findings from other human
studies (Petit et al., 2004) and indicate that aerobic exercise
might restore beneficial sleep EEG patterns. It is possible Frontiers in Systems Neuroscience Frontiers in Systems Neuroscience 05 frontiersin.org Cassim et al. 10.3389/fnsys.2022.855107 FIGURE 2
Proportion of the vigilance states derived from the baseline recordings (n = 18) and the ratio of the vigilance states between the baseline and
post-aerobic exercise recordings from five participants. (A) Distribution of vigilance states for all 18 baseline recordings. (B) Distribution of light
sleep (N1 + L2), deep sleep (N2 + N3) and REM sleep at baseline (n = 18). (C) Relative change of the vigilance states from baseline to
post-aerobic exercise recordings (n = 5). (D) Relative change of light sleep, deep sleep and REM sleep from baseline to post-aerobic exercise
recordings (n = 5). Sleep micro-architecture (A) For different levels of NREM sleep, i.e., N1, L2,
N2, and N3. (B) For light sleep and deep sleep. During deep sleep the power in the low (delta) frequencies was significantly higher and the
power in the high (beta) frequencies was significantly lower. Other
researchers
have
demonstrated
that
1
h
of
vigorous
exercise
can
increase
slow-wave
sleep
and
decrease wake transitions in a small study consisting of
young, healthy, and male subjects (Park et al., 2021). Our that these EEG changes could indicate a potential therapeutic
benefit of aerobic exercise on cognition and a potential non-
pharmacologic approach could mitigate age-related attenuation
of frontal EEG power during sleep. that these EEG changes could indicate a potential therapeutic
benefit of aerobic exercise on cognition and a potential non-
pharmacologic approach could mitigate age-related attenuation
of frontal EEG power during sleep. Frontiers in Systems Neuroscience 06 Frontiers in Systems Neuroscience frontiersin.org Cassim et al. 10.3389/fnsys.2022.855107 FIGURE 4
(A) Power spectral density (PSD) plots for deep sleep, both pre- (blue) and post- (orange) aerobic intervention (n = 5). A histogram for the
measured EEG power in each frequency bin is displayed. Solid lines represent average for all five participants and shading represents 95%
confidence interval. In deep sleep we see an increase in EEG power post-exercise, especially at moderate to high frequencies. (B) A comparison
of total non-REM EEG power pre- and post-aerobic intervention (n = 5). For non-REM sleep states (light and deep sleep), we see an increase in
total EEG power from pre- to post-aerobic intervention in frontal electrodes. Each pair of connected dots represents a single participant. (A) Power spectral density (PSD) plots for deep sleep, both pre- (blue) and post- (orange) aerobic intervention (n = 5). A histogram for the
measured EEG power in each frequency bin is displayed. Solid lines represent average for all five participants and shading represents 95%
confidence interval. In deep sleep we see an increase in EEG power post-exercise, especially at moderate to high frequencies. (B) A comparison
of total non-REM EEG power pre- and post-aerobic intervention (n = 5). For non-REM sleep states (light and deep sleep), we see an increase in
total EEG power from pre- to post-aerobic intervention in frontal electrodes. Each pair of connected dots represents a single participant. mechanisms are described in a recent review on the subject
(Santiago et al., 2022). Sleep micro-architecture results are particularly note worthy as they show benefits
for restorative sleep in older sedentary individuals with
exercise. Six months of exercise was shown to increase
sleep
efficiency
and
modulate
immune
biomarkers
in
previously sedentary elderly subjects (Abd El-Kader and
Al-Jiffri, 2019). Our 12-week regimen demonstrated benefit
in a similar population. While none of our participants
had a diagnosis of a pre-existing sleep disorder, these
are
often
under
diagnosed
and
more
common
as
we
age. It
has
been
shown
that
exercise
improves
sleep
quality
as
well
as
anti-depressive
response
in
patients
with chronic insomnia (Passos et al., 2014). It is not
unreasonable
to
assume
that
subclinical
disorders
of
mood
and/or
sleep
are
in
part
responsible
for
our
findings. Aging-related decreases in cortical inhibition might be
modified by 12-weeks of physical activity. Improvements in
restorative sleep may aid in titrating anesthetic regimens in
older participants. The loss of overall frontal EEG power
has been shown to be an important predictor of adverse
neurocognitive outcomes after surgery (Hesse et al., 2019). Therefore, it is conceivable that pre-habilitation with exercise
could be a viable option for at-risk patients scheduled for
surgery. Home-health devices as the one used in this study are a
practical way to evaluate sleep quality in older adults, and in the
future may be part of a comprehensive evaluation of cognitive
health. g
Although a relationship between low EEG power and
neurodegeneration has been established (Petit et al., 2004) it
remains difficult to point to an exact biological mechanism
that explains the benefits of exercise on sleep. It is known
that areas of the brain associated with learning and memory
(i.e., hippocampus) can increase in size with 12 weeks
of exercise in younger individuals (Nauer et al., 2020)
this was also associated with an enhanced performance on
memory tasks. It is not unreasonable to think that these
changes could also happen in elderly subjects. In fact,
16 weeks of aerobic exercise has been shown to prevent a
decrease in hippocampal subfield volume in elderly subjects
(Frodl et al., 2020). The potential for exercise to slow
neurodegeneration through enhanced activity and prevention of
synaptic and neuronal loss is an exciting avenue to investigate
to optimize cognitive health. Several putative molecular Frontiers in Systems Neuroscience Publisher’s note All claims expressed in this article are solely those of the
authors and do not necessarily represent those of their affiliated
organizations, or those of the publisher, the editors and the
reviewers. Any product that may be evaluated in this article, or
claim that may be made by its manufacturer, is not guaranteed
or endorsed by the publisher. Limitations We are aware of our limited sample size and hence,
further studies are needed to explore and interpret the
changes in sleep macro-architecture and EEG patterns that
are associated with an aerobic intervention. It remains
possible that exercise can results in substantial improvement
in perceptions of sleep quality in this population, however,
we
did
not
examine
any
subjective
self-evaluations
of
sleep
quality
in
our
participants. Some
researchers
may have concerns about the auto-scoring software for
classifying vigilance states. These devices show at least
80% agreement with expert raters and this is roughly
equivalent to the amount of variability between individual
raters (Levendowski et al., 2017). Frontiers in Systems Neuroscience 07 Frontiers in Systems Neuroscience frontiersin.org Cassim et al. 10.3389/fnsys.2022.855107 Conflict of interest The
data
analyzed
in
this
study
is
subject
to
the
following
licenses/restrictions:
The
raw
data
supporting the conclusions of this article will be made
available
by
the
authors,
without
undue
reservation. Requests
to
access
these
datasets
should
be
directed
to PG. The authors declare that the research was conducted in the
absence of any commercial or financial relationships that could
be construed as a potential conflict of interest. SUPPLEMENTARY TABLE 1 SUPPLEMENTARY TABLE 1 Statistical results corresponding to the figures. Statistical results corresponding to the figures. Ethics statement The studies involving human participants were reviewed
and approved by the Institutional Review Board, Emory School
of Medicine. The patients/participants provided their written
informed consent to participate in this study. SUPPLEMENTARY FIGURE 2 Power spectral density (PSD) plots for deep sleep, both pre- (blue) and
post- (orange) aerobic intervention (n = 5). A histogram for the
measured EEG power in each frequency bin is displayed. Solid lines
represent average for all five participants, and shading represents 95%
confidence interval. In deep sleep, we see an increase in EEG power
post-exercise, especially at moderate to high frequencies. In addition,
the AUC values and 95% bootstrapped CIs are presented. SUPPLEMENTARY FIGURE 1 Exemplary hypnogram of the classical vigilance states (A) and density
spectral array (B) of the pre-intervention EEG recording session. Author contributions The Supplementary Material for this article can be
found online at: https://www.frontiersin.org/articles/10.3389/
fnsys.2022.855107/full#supplementary-material TC: data quality, coordination, interpretation, analysis,
and draft and revision of the manuscript. KM and JN:
overall
project
design,
data
collection,
and
draft
and
revision
of
the
manuscript. VG,
CS,
and
MK:
data
analysis, and draft and revision of the manuscript. PG:
overall
project
design,
data
collection,
interpretation,
analysis,
and
draft
and
revision
the
manuscript. All
authors
contributed
to
the
article
and
approved
the
submitted version. Dzierzewski, J. M., Dautovich, N., and Ravyts, S. (2018). Sleep and Cognition in
Older Adults. Sleep Med. Clin. 13, 93–106. doi: 10.1016/j.jsmc.2017.09.009 Foley, D. J., Monjan, A. A., Brown, S. L., Simonsick, E. M., Wallace, R. B., and
Blazer, D. G. (1995). Sleep complaints among elderly persons: an epidemiologic
study of three communities. Sleep 18, 425–432. doi: 10.1093/sleep/18.6.425 Chennaoui, M., Arnal, P. J., Sauvet, F., and Leger, D. (2015). Sleep and exercise:
a reciprocal issue? Sleep Med. Rev. 20, 59–72. doi: 10.1016/j.smrv.2014.06.008 Carrier, J., Land, S., Buysse, D. J., Kupfer, D. J., and Monk, T. H. (2001). The
effects of age and gender on sleep EEG power spectral density in the middle years
of life (ages 20-60 years old). Psychophysiology 38, 232–242. Funding For light sleep and deep sleep. During deep sleep, the power in the low
(delta) frequencies was significantly higher and the power in the high
(beta) frequencies was significantly lower. In addition, the AUC values
and 95% bootstrapped CIs are presented. This work was supported by VA grants: IK2RX000956,
I01RX002619, IK2RX000744, I01RX002825, IK2BX001677, and
I50RX002358. Frodl, T., Strehl, K., Carballedo, A., Tozzi, L., Doyle, M., Amico, F., et al. (2020).
Aerobic exercise increases hippocampal subfield volumes in younger adults and References Effects of moderate-intensity exercise on polysomnographic and subjective
sleep quality in older adults with mild to moderate sleep complaints. J. Gerontol. A. Biol. Sci. Med. Sci. 63, 997–1004. doi: 10.1093/gerona/63.9.997 Petit, D., Gagnon, J. F., Fantini, M. L., Ferini-Strambi, L., and Montplaisir, J. (2004). Sleep and quantitative EEG in neurodegenerative disorders. J. Psychosom. Res. 56, 487–496. doi: 10.1016/j.jpsychores.2004.02.001 Kredlow, M. A., Capozzoli, M. C., Hearon, B. A., Calkins, A. W., and Otto,
M. W. (2015). The effects of physical activity on sleep: a meta-analytic review. J. Behav. Med. 38, 427–449. doi: 10.1007/s10865-015-9617-6 Sahlin, C., Franklin, K. A., Stenlund, H., and Lindberg, E. (2009). Sleep in
women: Normal values for sleep stages and position and the effect of age, obesity,
sleep apnea, smoking, alcohol and hypertension. Sleep Med. 10, 1025–1030. doi:
10.1016/j.sleep.2008.12.008 Kreuzer, M., Stern, M. A., Hight, D., Berger, S., Schneider, G., Sleigh, J. W.,
et al. (2020). Spectral and Entropic Features Are Altered by Age in the
Electroencephalogram in Patients under Sevoflurane Anesthesia. Anesthesiology
132, 1003–1016. doi: 10.1097/ALN.0000000000003182 Santiago, J. A., Quinn, J. P., and Potashkin, J. A. (2022). Physical Activity
Rewires the Human Brain against Neurodegeneration. Int. J. Mol. Sci. 23:6223. doi: 10.3390/ijms23116223 Lariviere, M., Goffaux, P., Marchand, S., and Julien, N. (2007). Changes in pain
perception and descending inhibitory controls start at middle age in healthy adults. Clin. J. Pain 23, 506–510. doi: 10.1097/AJP.0b013e31806a23e8 Schulz,
H. (2008). Rethinking
sleep
analysis. J. Clin. Sleep
Med. 4, 99–103. Levendowski, D. J., Ferini-Strambi, L., Gamaldo, C., Cetel, M., Rosenberg,
R., and Westbrook, P. R. (2017). The Accuracy, Night-to-Night Variability, and
Stability of Frontopolar Sleep Electroencephalography Biomarkers. J. Clin. Sleep
Med. 13, 791–803. doi: 10.5664/jcsm.6618 Scullin, M. K. (2013). Sleep, memory, and aging: the link between slow-wave
sleep and episodic memory changes from younger to older adults. Psychol. Aging
28, 105–114. doi: 10.1037/a0028830 Scullin, M. K., and Bliwise, D. L. (2015). Sleep, cognition, and normal aging:
integrating a half century of multidisciplinary research. Perspect. Psychol. Sci. 10,
97–137. doi: 10.1177/1745691614556680 Mandrekar,
J. N. (2010). Receiver
operating
characteristic
curve
in
diagnostic test assessment. J. Thorac. Oncol. 5, 1315–1316. doi: 10.1097/JTO. 0b013e3181ec173d Silber, M. H., Ancoli-Israel, S., Bonnet, M. H., Chokroverty, S., Grigg-
Damberger, M. M., Hirshkowitz, M., et al. (2007). The visual scoring of sleep in
adults. J. Clin. Sleep Med. 3, 121–131. McGregor, K. M., Crosson, B., Krishnamurthy, L. C., Krishnamurthy, V.,
Hortman, K., Gopinath, K., et al. (2018). References Abd El-Kader, S. M., and Al-Jiffri, O. H. (2019). Aerobic exercise modulates
cytokine profile and sleep quality in elderly. Afr. Health Sci. 19, 2198–2207. doi:
10.4314/ahs.v19i2.45 Carrier, J., Land, S., Buysse, D. J., Kupfer, D. J., and Monk, T. H. (2001). The
effects of age and gender on sleep EEG power spectral density in the middle years
of life (ages 20-60 years old). Psychophysiology 38, 232–242. Carrier, J., Land, S., Buysse, D. J., Kupfer, D. J., and Monk, T. H. (2001). The
effects of age and gender on sleep EEG power spectral density in the middle years
of life (ages 20-60 years old). Psychophysiology 38, 232–242. Chennaoui, M., Arnal, P. J., Sauvet, F., and Leger, D. (2015). Sleep and exercise:
a reciprocal issue? Sleep Med. Rev. 20, 59–72. doi: 10.1016/j.smrv.2014.06.008 Berres, S., and Erdfelder, E. (2021). The sleep benefit in episodic memory: An
integrative review and a meta-analysis. Psychol. Bull. 147, 1309–1353. doi: 10.1037/
bul0000350 Dzierzewski, J. M., Dautovich, N., and Ravyts, S. (2018). Sleep and Cognition in
Older Adults. Sleep Med. Clin. 13, 93–106. doi: 10.1016/j.jsmc.2017.09.009 Bliwise, D. L., King, A. C., Harris, R. B., and Haskell, W. L. (1992). Prevalence
of self-reported poor sleep in a healthy population aged 50-65. Soc. Sci. Med. 34,
49–55. doi: 10.1016/0277-9536(92)90066-y Foley, D. J., Monjan, A. A., Brown, S. L., Simonsick, E. M., Wallace, R. B., and
Blazer, D. G. (1995). Sleep complaints among elderly persons: an epidemiologic
study of three communities. Sleep 18, 425–432. doi: 10.1093/sleep/18.6.425 Borbely, A. A., Daan, S., Wirz-Justice, A., and Deboer, T. (2016). The two-
process model of sleep regulation: a reappraisal. J. Sleep Res. 25, 131–143. doi:
10.1111/jsr.12371 Borbely, A. A., Daan, S., Wirz-Justice, A., and Deboer, T. (2016). The two-
process model of sleep regulation: a reappraisal. J. Sleep Res. 25, 131–143. doi:
10.1111/jsr.12371 Frodl, T., Strehl, K., Carballedo, A., Tozzi, L., Doyle, M., Amico, F., et al. (2020). Aerobic exercise increases hippocampal subfield volumes in younger adults and 08 Frontiers in Systems Neuroscience Frontiers in Systems Neuroscience frontiersin.org Cassim et al. 10.3389/fnsys.2022.855107 and enhances memory in young adults. Hippocampus 30, 488–504. doi: 10.1002/
hipo.23166 prevents volume decline in the elderly. Brain Imag. Behav. 14, 1577–1587. doi:
10.1007/s11682-019-00088-6 Grober, E., and Sliwinski, M. (1991). Development and validation of a model for
estimating premorbid verbal intelligence in the elderly. J. Clin. Exp. Neuropsychol. 13, 933–949. doi: 10.1080/01688639108405109 Nocera, J., Crosson, B., Mammino, K., and McGregor, K. References M. (2017). Changes
in Cortical Activation Patterns in Language Areas following an Aerobic Exercise
Intervention in Older Adults. Neural. Plast 2017:6340302. doi: 10.1155/2017/
6340302 Hesse, S., Kreuzer, M., Hight, D., Gaskell, A., Devari, P., Singh, D., et al. (2019). Association of electroencephalogram trajectories during emergence from
anaesthesia with delirium in the postanaesthesia care unit: an early sign of
postoperative complications. Br. J. Anaesth. 122, 622–634. doi: 10.1016/j.bja.2018. 09.016 Panagiotou, M., Papagiannopoulos, K., Rohling, J. H. T., Meijer, J. H., and
Deboer, T. (2018). How Old Is Your Brain? Slow-Wave Activity in Non-rapid-eye-
movement Sleep as a Marker of Brain Rejuvenation After Long-Term Exercise in
Mice. Front. Aging Neurosci. 10:233. doi: 10.3389/fnagi.2018.00233 Heyn, P., Abreu, B. C., and Ottenbacher, K. J. (2004). The effects of exercise
training on elderly persons with cognitive impairment and dementia: a meta-
analysis. Arch. Phys. Med. Rehabil. 85, 1694–1704. doi: 10.1016/j.apmr.2004.03. 019 Papalambros, N. A., Santostasi, G., Malkani, R. G., Braun, R., Weintraub, S.,
Paller, K. A., et al. (2017). Acoustic Enhancement of Sleep Slow Oscillations
and Concomitant Memory Improvement in Older Adults. Front. Hum. Neurosci. 11:109. doi: 10.3389/fnhum.2017.00109 Hillman, C. H., Erickson, K. I., and Kramer, A. F. (2008). Be smart, exercise
your heart: exercise effects on brain and cognition. Nat. Rev. Neurosci. 9, 58–65. doi: 10.1038/nrn2298 Park, I., Diaz, J., Matsumoto, S., Iwayama, K., Nabekura, Y., Ogata, H., et al. (2021). Exercise improves the quality of slow-wave sleep by increasing slow-wave
stability. Sci. Rep. 11:4410. doi: 10.1038/s41598-021-83817-6 Hotting, K., and Roder, B. (2013). Beneficial effects of physical exercise on
neuroplasticity and cognition. Neurosci. Biobehav. Rev. 37, 2243–2257. doi: 10. 1016/j.neubiorev.2013.04.005 Passos, G. S., Poyares, D., Santana, M. G., Teixeira, A. A., Lira, F. S., Youngstedt,
S. D., et al. (2014). Exercise improves immune function, antidepressive response,
and sleep quality in patients with chronic primary insomnia. Biomed. Res. Int. 2014:498961. doi: 10.1155/2014/498961 Kaliman, P., Parrizas, M., Lalanza, J. F., Camins, A., Escorihuela, R. M., and
Pallas, M. (2011). Neurophysiological and epigenetic effects of physical exercise
on the aging process. Ageing Res. Rev. 10, 475–486. doi: 10.1016/j.arr.2011.05.002 Paul-Savoie, E., Marchand, S., Morin, M., Bourgault, P., Brissette, N.,
Rattanavong, V., et al. (2012). Is the deficit in pain inhibition in fibromyalgia
influenced by sleep impairments? Open Rheumatol. J. 6, 296–302. doi: 10.2174/
1874312901206010296 King, A. C., Pruitt, L. A., Woo, S., Castro, C. M., Ahn, D. K., Vitiello, M. V., et al. (2008). Zhong, H. H., Yu, B., Luo, D., Yang, L. Y., Zhang, J., Jiang, S. S., et al. (2019).
Roles of aging in sleep. Neurosci. Biobehav. Rev. 98, 177–184. doi: 10.1016/j.
neubiorev.2019.01.013 References Effects of a 12-Week Aerobic Spin
Intervention on Resting State Networks in Previously Sedentary Older Adults. Front. Psychol. 9:2376. doi: 10.3389/fpsyg.2018.02376 Vitiello, M. V., Larsen, L. H., and Moe, K. E. (2004). Age-related sleep change:
Gender and estrogen effects on the subjective-objective sleep quality relationships
of healthy, noncomplaining older men and women. J. Psychosom. Res. 56, 503–510. doi: 10.1016/S0022-3999(04)00023-6 McGregor, K. M., Crosson, B., Mammino, K., Omar, J., Garcia, P. S., and
Nocera, J. R. (2017). Influences of 12-Week Physical Activity Interventions on
TMS Measures of Cortical Network Inhibition and Upper Extremity Motor
Performance in Older Adults-A Feasibility Study. Front. Aging Neurosci. 9:422. doi: 10.3389/fnagi.2017.00422 Voss, M. W., Heo, S., Prakash, R. S., Erickson, K. I., Alves, H., Chaddock,
L., et al. (2013). The influence of aerobic fitness on cerebral white matter
integrity and cognitive function in older adults: results of a one-year
exercise intervention. Hum. Brain Mapp. 34, 2972–2985. doi: 10.1002/hbm. 22119 Nasreddine, Z. S., Phillips, N. A., Bedirian, V., Charbonneau, S., Whitehead,
V., Collin, I., et al. (2005). The Montreal Cognitive Assessment, MoCA: a brief
screening tool for mild cognitive impairment. J. Am. Geriatr. Soc. 53, 695–699. doi: 10.1111/j.1532-5415.2005.53221.x Zhong, H. H., Yu, B., Luo, D., Yang, L. Y., Zhang, J., Jiang, S. S., et al. (2019). Roles of aging in sleep. Neurosci. Biobehav. Rev. 98, 177–184. doi: 10.1016/j. neubiorev.2019.01.013 Nauer, R. K., Dunne, M. F., Stern, C. E., Storer, T. W., and Schon, K. (2020). Improving fitness increases dentate gyrus/CA3 volume in the hippocampal head 09 Frontiers in Systems Neuroscience Frontiers in Systems Neuroscience frontiersin.org
|
https://openalex.org/W2799903271
|
https://www.repository.cam.ac.uk/bitstream/1810/276538/4/c8fd00042e.pdf
|
English
| null |
Predicting the structures and associated phase transition mechanisms in disordered crystals <i>via</i> a combination of experimental and theoretical methods
|
Faraday discussions
| 2,018
|
cc-by
| 8,417
|
View Article Online
View Journal View Article Online
View Journal View Article Online
View Journal This article can be cited before page numbers have been issued, to do this please use: M. T. Ruggiero, J. Kölbel, Q. Li and
J. A. Zeitler, Faraday Discuss., 2018, DOI: 10.1039/C8FD00042E. Faraday
Discussions Accepted Manuscript
Accepted Manuscript Accepted Manuscript
Accepted Manuscript This manuscript will be presented and discussed at a forthcoming Faraday Discussion meeting. All delegates can contribute to the discussion which will be included in the final volume. Register now to attend! Full details of all upcoming meetings: http://rsc.li/fd-upcoming-m This is an Accepted Manuscript, which has been through the
Royal Society of Chemistry peer review process and has been
accepted for publication. This is an Accepted Manuscript, which has been through the
Royal Society of Chemistry peer review process and has been
accepted for publication. Faraday
Discussions
Royal Society of
Chemistry Faraday
Discussions Accepted Manuscripts are published online shortly after
acceptance, before technical editing, formatting and proof reading. Using this free service, authors can make their results available
to the community, in citable form, before we publish the edited
article. We will replace this Accepted Manuscript with the edited
and formatted Advance Article as soon as it is available. Accepted Manuscripts are published online shortly after
acceptance, before technical editing, formatting and proof reading. Using this free service, authors can make their results available
to the community, in citable form, before we publish the edited
article. We will replace this Accepted Manuscript with the edited
and formatted Advance Article as soon as it is available. Faraday
Discussions You can find more information about Accepted Manuscripts in the
Information for Authors.
You can find more information about Accepted Manuscripts in the
Information for Authors. You can find more information about Accepted Manuscripts in the
Information for Authors. You can find more information about Accepted Manuscripts in the
Information for Authors. a Department of Chemical Engineering and Biotechnology,
University of Cam-
bridge, Philippa Fawcett Drive, Cambridge, CB3 0AS, United Kingdom; E-mail:
mtr34@cam.ac.uk Information for Authors.
Information for Authors. Please note that technical editing may introduce minor changes
to the text and/or graphics, which may alter content. The journal’s
standard Terms & Conditions and the Ethical guidelines still
apply. In no event shall the Royal Society of Chemistry be held
responsible for any errors or omissions in this Accepted Manuscript
or any consequences arising from the use of any information it
contains. Please note that technical editing may introduce minor changes
to the text and/or graphics, which may alter content. The journal’s
standard Terms & Conditions and the Ethical guidelines still
apply. In no event shall the Royal Society of Chemistry be held
responsible for any errors or omissions in this Accepted Manuscript
or any consequences arising from the use of any information it
contains. www.rsc.org/faraday_d www.rsc.org/faraday_d Page 1 of 9 Faraday Discussions Faraday Discussions View Article Online
DOI: 10.1039/C8FD00042E Michael T. Ruggiero,∗a Johanna Kölbel,a Qi Li,a and J. Axel Zeitlera Disordered materials make up a large portion of condensed phase systems, but the difficulties in
describing their structures and molecular dynamics limit their potential applications. Disordered
crystalline systems, also known as plastic crystals, offer a unique perspective into these factors
because the system retains a degree of crystallinity, reducing the degrees of freedom that must be
explored when interpreting the results. However, while disordered crystals do diffract X-rays, it is
difficult to fully resolve a meaningful crystalline structure, with the best scenario resulting in lattice
parameters. In this study, we use a combination of experimental terahertz time-domain spec-
troscopy, and theoretical solid-state ab initio density functional theory and molecular dynamics
simulations to fully elucidate the structures and associated dynamics of organic molecular solids. The results highlight that this combination provides a complete description of the energetic and
mechanistic pathways involved in the formation of disordered crystals, and highlights the impor-
tance of low-frequency dynamics on their properties. Finally, with structures fully determined and
validated by the experimental results, recent progress into anharmonic calculations, namely the
quasi-harmonic approximation method, enables full temperature and pressure-dependent prop-
erties to be understood within the framework of the potential energy hyper-surface structure. Faraday Discussions A Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36.
This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. y Discussions Accepted Manuscrip Michael T. Ruggiero,∗a Johanna Kölbel,a Qi Li,a and J. Axel Zeitlera Predicting the Structures and Associated Phase Tran-
sition Mechanisms in Disordered Crystals via a Com-
bination of Experimental and Theoretical Methods cussions Accepted Manuscript Faraday Discussions Accepted Manuscript Accepted Manuscript 1
Introduction molecular potential energy surfaces being probed simultaneously,
providing a valuable measure of the bulk structure and dynamics. Terahertz time-domain spectroscopy (THz-TDS) is the far-IR ex-
tension of the more traditional FTIR technique, but unlike FTIR,
which probes internal vibrations, there are no standard func-
tional group specific terahertz vibrations, meaning each solid has
a unique spectral fingerprint at terahertz frequencies.9,10 There-
fore, the interpretation of the THz-TDS spectra requires signifi-
cant computational modelling, and (at a minimum) the structures
of the associated solids are necessary for initialising the simula-
tions.11,12 Disordered (amorphous) solids are an important class of materi-
als that have found use in a wide range of fields, from the semi-
conductor1 to pharmaceutical industries.2 The utility of amor-
phous materials lies in their metastable nature,3,4 which ulti-
mately leads to increased molecular dynamics compared to their
crystalline counterparts.5 However, the actual structures and as-
sociated motions of the molecules within disordered solids remain
a topic of debate, as traditional techniques, such as X-ray crys-
tallography, provide little information that can be directly inter-
preted without significant approximations.6 Recently, the importance of low-frequency (≤330cm−1 or
10THz) collective vibrations on bulk properties and phase trans-
formation phenomena in molecular solids has been suggested,7
however the types of motions occurring in disordered materials
have not yet been completely understood. Experimentally, low-
frequency (terahertz) vibrational spectroscopy is a powerful tool
for investigating solids, as the terahertz radiation is able to excite
large-amplitude collective motions of entire molecules within the
solid.8 This leads to both the internal conformational and inter- Disordered (or plastic) crystals are a class of materials that ex-
hibit pseudo-order, in that the molecules occupy crystalline sites
but lack true translational symmetry due to uncoupled motions
within their crystallographic sites.13,14 This leads to a scenario
where the material diffracts X-rays (like a traditional crystal), but
rather than yielding meaningful atomic-level structural details the
data only provides a broad description of occupied sites within the
solid, i.e. Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36.
This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. araday Discussions Accepted Man AIMD simulations also enable structural transformations to be
investigated.28–30 As many plastic crystals often undergo a trans-
formation to an ordered crystal at low temperatures, it is possible
to analyse the ordered form experimentally with diffraction meth-
ods to obtain an atomic-level structure. By using this structure as
a starting point for the AIMD simulations, the prediction of both
the transformation pathway as well as the disordered structure
can be achieved. Faraday Discussions Accep 1
Introduction an average “sphere" of electron density at each lattice
site.15 However, the psuedo-order limits the possible degrees of
freedom that the molecules within the solid have, enabling more
detailed investigation into the bulk structure and dynamics to be
performed.16,17 Because disordered crystals are ultimately disordered, diffrac- Journal Name, [year], [vol.],
1–9 | 1 Faraday Discussions Page 2 of 9 Page 2 of 9 Page 2 of 9 View Article Online
DOI: 10.1039/C8FD00042E tion methods often rely on other techniques to fully elucidate
the static-ordered structures.18 While this has been successfully
achieved in a number of cases, the static picture of a disordered
crystal is not truly physical, since the molecules are free to move
in their crystallographic sites uncoupled from their neighbours.19
In order to obtain a more physical description, molecular dynam-
ics (MD) simulations can be used as a means of predicting the
nature of the motions occurring in the solid,20,21 as well as pre-
dicting the vibrational spectra through determination of either
the molecular dipole moments (for infrared (IR) spectroscopies),
or the molecular polarisabilities (for Raman spectroscopies).22,23
However, traditional MD simulations are performed using user-
defined force constants, which leads to biased vibrational results. A solution to this is ab initio molecular dynamics (AIMD), in
which the forces are recalculated using density functional theory
(DFT) at each individual time-step, leading to a more accurate
description of the vibrational dynamics occurring within the sys-
tem.24–27 2 |
1–9
Journal Name, [year], [vol.], 2.1
Experimental Methods Parachlorobenzamide was purchased from Sigma-Aldrich (98%,
Poole, UK) and was used as-received. Samples were prepared for
THz-TDS measurements by mixing the sample with polyethylene
to a 10% w/w concentration and subsequently grinding with a
mortar and pestle to homogenise the sample and reduce particle
size. A hydraulic press was then used to press the mixture into
a 13mm diameter freestanding pellet with a thickness of 2mm. A corresponding pellet containing pure polyethylene was also
made to act as a standard blank for absorption measurements. All experimental THz-TDS spectra were obtained using a com-
mercial TeraPulse 4000 spectrometer (TeraView Ltd., Cambridge,
UK). Variable-temperature measurements were performed using
a liquid nitrogen cryostat (Janis, Massachusetts, USA) equipped
with a externally controlled heating element (Lakeshore 330,
Ohio, USA), permitting measurements from 80−400K. The time-
domain waveforms were symmetrically zero-padded and Fourier
transformed to yield terahertz power spectra. The absorption
spectra presented are a result of division of the sample spectra
by that of a blank spectrum. Discussions Accepted Manuscript ns Accepted Manuscript Faraday Discussions Accepted Manuscript 3.1
Terahertz Time-Domain Spectroscopy In order to determine the vibrational response of PCB over a
wide temperature range, THz-TDS spectra were acquired from
100 −350K and the results are shown in Figure 2. At low tem-
peratures, there is at least one distinct feature at 1.23THz, with a
number of potential features present at higher frequencies, which
is characteristic of a well-ordered crystalline solid. As the tem-
perature is increased, the main absorption feature significantly
broadens and shifts to lower frequencies, likely due to thermal ex-
pansion leading to a softening of the vibrational potential energy
and an increased number of thermally-activated vibrational relax-
ation pathways, the latter of which leads to spectral broadening. It is important to note that while many materials experience these
phenomena upon heating, the magnitude of the broadening ob-
served here is rather large in comparison to most molecular crys-
tals.27,55 The origins of the spectral broadening might be related
to the ordered solid deviating from an ideal crystalline geometry,
as minor displacements from equilibrium positions would lead
to slightly different vibrational potentials for each unit molecule. This would manifest in the experimental spectra as broadened
features, as observed in the continuous phase transition of crys-
talline N-methyl-4-carboxypyridinium chloride.7 Additionally, a
decrease in the intensity of the spectra can also be attributed to
the increased vibrational motion with increasing temperature, as
the IR intensity is proportional to the change in dipole moment
with respect to the atomic displacement, I ∝∂µ
∂Q.27 Overall, it is
likely that the experimental spectral broadening is due to a com-
bination of all of these phenomena, which results in the large
effect observed. Faraday Discussions Fig. 2 Experimental THz-TDS spectra of PCB at various temperatures
ranging from 100K (blue) to 350K (red). There is a marked transition
between 310K and 320K, corresponding to the order-disorder transition,
with all spectra corresponding to the α-form coloured blue, while all spec-
tra corresponding to the γ-form coloured red. The inset shows the ab-
sorption at 1.5THz as a function of temperature, with the experimental
transition temperature of 317K noted by a dotted vertical line. Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36.
This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. y Discussions Accepted Manuscrip Faraday Discussions Accepted Man 3
Results and Discussion Faraday Discussions Acce Fig. 2 Experimental THz-TDS spectra of PCB at various temperatures
ranging from 100K (blue) to 350K (red). There is a marked transition
between 310K and 320K, corresponding to the order-disorder transition,
with all spectra corresponding to the α-form coloured blue, while all spec-
tra corresponding to the γ-form coloured red. The inset shows the ab-
sorption at 1.5THz as a function of temperature, with the experimenta
transition temperature of 317K noted by a dotted vertical line. Faraday Discussions Page 3 of 9 View Article Online tra exhibiting a noticeable increase in intensity across the entire
spectral region that continues to increase with rising temperature,
a common trait of disordered solids and in contrast to crystalline
solids, which typically undergo a decrease in absorption as tem-
perature is increased. While there is apparently little difference
in the shape of the individual spectra (i.e. both spectra appear
to be featureless with an increasing baseline), there is an obvious
difference in the temperature-dependence of the spectra. This is
highlighted in the Figure 2 inset, which shows the clear change
in the absorption coefficient at 1.5THz that is observed between
310K and 320K. This discontinuity in the spectral trends corre-
sponds to the order-disorder phase transition in PCB, which is
in good agreement with the published transition temperature of
317K.32,33,56 However, unlike fully-disordered materials, which
do not exhibit any resolvable low-frequency vibrational features,
the spectra of the disordered γ-PCB solid seemingly exhibits at
least one absorption at ∼1.72THz. It is important to note that ex-
perimentally the discontinuity in the THz-TDS measurements is
reproducible upon heating and cooling, indicating that the kinet-
ics of the transformation are faster than the experimental method
can probe (minutes, due to the equilibration time required for
temperature changes). DOI: 10.1039/C8FD00042E tra exhibiting a noticeable increase in intensity across the entire
spectral region that continues to increase with rising temperature,
a common trait of disordered solids and in contrast to crystalline
solids, which typically undergo a decrease in absorption as tem-
perature is increased. While there is apparently little difference
in the shape of the individual spectra (i.e. both spectra appear
to be featureless with an increasing baseline), there is an obvious
difference in the temperature-dependence of the spectra. This is
highlighted in the Figure 2 inset, which shows the clear change
in the absorption coefficient at 1.5THz that is observed between
310K and 320K. This discontinuity in the spectral trends corre-
sponds to the order-disorder phase transition in PCB, which is
in good agreement with the published transition temperature of
317K.32,33,56 However, unlike fully-disordered materials, which
do not exhibit any resolvable low-frequency vibrational features,
the spectra of the disordered γ-PCB solid seemingly exhibits at
least one absorption at ∼1.72THz. Faraday Discussions It is important to note that ex-
perimentally the discontinuity in the THz-TDS measurements is
reproducible upon heating and cooling, indicating that the kinet-
ics of the transformation are faster than the experimental method
can probe (minutes, due to the equilibration time required for
temperature changes). DOI: 10.1039/C8FD00042E 2.2.2
S-DFT Simulations All s-DFT simulations were performed using the fully-periodic
CRYSTAL17 software package.46 In order to be comparable to
the AIMD simulations, the computational parameters were kept
as similar as possible between the two methods. The atomic or-
bitals were represented using the atom-centred def2-SVP basis
set,47 and the D3-corrected PBE density functional was used for
all calculations. The experimental α-PCB structure32,33 and the
AIMD-generated unit cell of the γ-form (vide infra) were used
to initialise the respective simulations. In both cases the simu-
lations were performed in the absence of any space group sym-
metry in order to mimic the AIMD methodology and remove
any spurious constraints. Both structures were fully optimised
to a convergence of ∆E ≤10−8 hartree, and vibrational analy-
ses were performed using numerical differentiation within the
harmonic approximation, with an energy convergence of ∆E ≤
10−11 hartree.48,49 Infrared intensities were determined using the
Berry phase approach.50 Quasi-harmonic approximation (QHA)
simulations were performed by simulating the optimised struc-
tures and vibrational properties at four volumes corresponding
to a −2%,0,+2%, and +4% contraction/expansion relative to the
equilibrium structure.51–54 ns Accepted Manuscript Faraday Discussions Accepted Manuscript 2.2
Computational Methods
2.2.1
AIMD Simulations All AIMD simulations were performed using the freely-available
CP2K software package,34,35 which incorporates periodic bound-
ary conditions. A 1 × 3 × 2 supercell (36 molecules, 576 atoms)
was generated from the experimental structure of α-PCB in or-
der to minimise interactions with neighbouring unit cells and
fully capture the physical phase transformation process. Simu-
lations were performed within the isothermal-isobaric (NPT) en-
semble, with the temperature controlled using a Nosé-Hoover
chain thermostat,36,37 and pressure maintained using the ex-
tended Nosé-Hoover barostat of Martyna et al.38,39 All atoms
were modelled using the double-ζ DZVP-GTH basis set40 and the
Goedecker-Teter-Hutter (GTH) pseudopotentials.41 The Perdew-
Burke-Ernzerhof (PBE) density functional42 and Grimme D3-
dispersion correction43,44 were used for all AIMD simulations. A
triclinic cell was chosen so that all lattice components were free to
relax throughout the simulations. A 1.0fs time-step was used for
all simulations, and each trajectory was run for a total of 20 000fs
(20ps). The convergence criteria were set to ∆E ≤10−9 hartree
for all simulations. In this study, a combination of experimental THz-TDS and the-
oretical static solid-state-DFT (s-DFT) and AIMD simulations are
used to predict the structures, dynamics, and phase-transition
pathways in the order-disorder transformation of parachloroben-
zamide (PCB, Figure 1).31 The static structures of both the
ordered and plastic phases of the solid have been previously
reported experimentally using synchrotron-based X-ray diffrac-
tion,32,33 which enables the proposed methodology to be vali-
dated by experimental results. Overall, this combination of tech-
niques allows the static structures, as well as the associated dy-
namical aspects, to be fully understood, providing insight into
the energetic and mechanistic forces that drive the order-disorder
phase transition in plastic crystals. Fig. 1 Skeletal and three-dimensional models of a single PCB molecule. IR-spectra were generated by performing NVT simulations
starting from the equilibrated NPT supercells. A time-step of 0.5fs
was used for all vibrational analyses, for a total trajectory length
of 30ps. Both solids underwent an initial equilibrium period prior
to the production simulation, which was set to 3.0ps and 5.0ps
for the α- and γ-forms, respectively. The molecular dipoles were
determined via calculation of the Wannier centres, which was per-
formed every 2.5fs. The resulting trajectories were analysed using
the TRAVIS post-processing software package,45 and IR-spectra
were calculated by performing the Fourier transform of the dipole
moment autocorrelation function.24,27 Fig. 1 Skeletal and three-dimensional models of a single PCB molecule. Faraday Discussions Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36.
This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. aday Discussions Accepted Man Faraday Discussions Acce In the α-form, there are a number of low-frequency vibra-
tions predicted that result in good agreement with the experi-
mental spectrum. Overall, the predicted spectrum is slightly over-
estimated in terms of frequency, which is likely due to the simula-
tions being performed at 0K while the experimental spectrum was
recorded at 80K, as in some cases it has been shown that spectra
continue to shift and sharpen upon cooling to temperatures as
low at 4K.57 Indeed, the s-DFT simulations are also blue-shifted
compared to the 100K AIMD simulations, further indicating that
the spectrum is strongly influenced by temperature. Addition-
ally, AIMD vibrational analysis performed at higher temperatures
confirms that the spectra become much more diffuse and red-
shifted as the transition temperature is approached (Figure 6). The temperature-dependent AIMD-simulated vibrational spectra
generally reproduce the experimental results, predicted a red-
shift of most features, as well as an increase in absorption in-
tensity between ∼1.25THz and 1.85THz, which originates from
the combination of spectral shifting and broadening. Faraday Discussions Six molecules (two stacked rows of three molecules each, with
alternating dipole moments) were extracted from the equilibrated
AIMD simulation and a unit cell was fit around them. The system
was loaded into the fully-periodic CRYSTAL17 software package
and optimised within the constraints of the input lattice vectors,
in order to minimise any possibility of relaxation back into the α-
form. Upon successful optimisation, the system was then allowed
to fully relax with no constraints (lattice vectors and atomic posi-
tions). The resulting structure (Figure 3) was found to be triclinic
with P1 space group symmetry, lattice parameters of a = 4.934
Å, b = 5.383 Å, c = 13.794 Å, α = 93.931◦, β = 116.421◦, and
γ = 93.498◦, and a Z = 2 (Z′ = 1). This structure is in good gen-
eral agreement with the previously suggested static-structure of
γ-PCB (bond, angle, and dihedral angle RMSDs of 0.035 Å, 0.656◦,
and 2.552◦, respectively), although with slightly contracted lattice
vectors due to the s-DFT simulations effectively simulating a 0K
crystal. The vibrational mode-types are mainly hindered rotations of
the individual molecules with respect to one another, along both
the long molecular axis as well as librations orthogonal to the
plane of the molecule, with the exception of the weak predicted
feature at 1.645THz which is a translational-type motion. Faraday Discussions Page 4 of 9 View Article Online the plastic crystal, in which the molecules generally move freely
within their sites, there is a clear correlated rocking-type motion
present. Interestingly, these oscillations have periods of about
1 −2ps, which is on the order of the period of terahertz vibra-
tions (1THz = 1012 Hz = 1ps). With the structures of the two
forms known, a more rigorous analysis of the vibrational dynam-
ics occurring within the solids can be performed. DOI: 10.1039/C8FD00042E pressure set to 1 bar and the temperature varied from 100−400K,
with a time-step of 1.0fs. In the high temperature simulations
(above 300K), the system quickly undergoes a large-scale trans-
formation, with all of the PCB molecules experiencing large am-
plitude hindered-rotational type motions about their long axis
(see Figure 3). It is important to note that the AIMD simula-
tions underestimate the transition temperature by ∼20K, how-
ever, given the limitations of the exact implementation of thermal
energy into the model, this represents a relatively minor error
(20K ≈0.04kcal mol−1). 4 |
1–9
Journal Name, [year], [vol.], 3.3
Vibrational Dynamics Accepted Manuscript 3.2
Structural Elucidation and Phase Transformation Path-
way In order to determine the structure, and associated dynamics, of
the disordered γ-PCB solid, AIMD simulations were performed. Initially, a 1 × 3 × 2 supercell of the α-PCB solid was generated
and used as the starting geometry for the AIMD simulations. Fully
periodic NPT simulations were performed using CP2K, with the Upon increasing the temperature above 310K, there is a clear
change in the vibrational response of the material, with the spec- Journal Name, [year], [vol.],
1–9 | 3 Faraday Discussions Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36.
This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. Faraday Discussions Accepted Manuscript Discussions Accepted Manuscript The vibrational dynamics of both PCB solids were predicted using
the AIMD simulations, and the results are shown in Figure 5. In
both cases, the AIMD simulations generally reproduce the exper-
imental spectra, yielding a number of features that correspond to
those experimentally observed. However, the AIMD spectra pro-
vide little specific atomic-level details surrounding the nature of
each individual vibrational mode, leaving further analyses nec-
essary. In this regard, s-DFT simulations are able to determine
the precise atomic-motions that produce each vibrational tran-
sition, and this method was employed once the static structure
of the γ-form was determined. The static crystal structures of
both forms of PCB were fully optimised with no constraints using
CRYSTAL17, and subsequently the phonon properties were cal-
culated using a finite difference method within the harmonic ap-
proximation. The results of the vibrational simulations are shown
in Figure 5 for both crystals. The phase transformation can be clearly observed through
analysis of the angle between the planes formed by the two dis-
tinct molecular environments in the α-form. In low-temperature
AIMD simulations (below 300K) the molecules do not undergo
any large-scale rearrangement, rather they simply oscillate within
their crystallographic sites about their equilibrium positions, typ-
ical of an ordered molecular crystal. However, at higher tem-
peratures, there still exist oscillations, but with a much larger
magnitude compared to the low-temperature case, as well as an
overall trend towards a vanishing angle between the planes of
the molecule (i.e. all molecules trending towards forming pla-
nar sheets). Additionally, the increased molecular motion in the
high-temperature simulation results in an initial expansion of the
simulation cell, which is a direct example of vibrational dynamics
leading to thermal expansion. After ∼10ps, the high-temperature
simulation relaxes to a new bulk structure that generally exhibits
an overall planar arrangement of all of the molecules within
the system, however the individual molecules continue to un-
dergo rather large-amplitude hindered-rotational type motions
that are seemingly uncoupled from the bulk. Moreover, in the
high-temperature simulation, the molecules are generally not as
well-ordered as in the low-temperature simulation, with many de-
viations from the static crystal structure readily apparent. How-
ever, there do exist a number of local ordering events that makes
extracting a molecular basis possible, and this process was used
to generate a crystallographic unit cell for the γ-form of PCB. Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36.
This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. Furthermore, the observation of this mode experi-
mentally confirms that this vibration is not a long-range collective
vibration, which would lead to a very broad experimental absorp-
tion, but rather is predominantly a local effect, being primarily
dependent on a single molecular neighbour. Fig. 4 AIMD simulated angles between adjacent molecular planes at
100K (blue) and 300K (red). A horizontal dashed line is drawn through
the origin as a guide. Faraday Discussions Accep However, the s-DFT predicted spectrum fails to reproduce the
correct IR density of states in the terahertz region as shown by
the experimental and AIMD results. This is not surprising, as
the static model is not a true representation of the disordered
solid. However, there are a number of IR-inactive modes pre-
dicted which could become activated upon removal of crystalline
symmetry, and interestingly these transitions are predicted at
similar frequencies to those predicted by the AIMD simulations. These modes, shown in Figure 5 as dotted sticks, also corre-
spond to hindered-rotational motions, with the lowest-frequency
mode (1.296THz) corresponding well with the mode type of the
1.337THz vibration in the α-form. Indeed, the occurrence of a
transition in both the experimental and AIMD-predicted spectrum
at ∼1.17THz confirms that in the absence of true-crystalline sym-
metry this vibration can become activated. Faraday Discussions Fig. 4 AIMD simulated angles between adjacent molecular planes at
100K (blue) and 300K (red). A horizontal dashed line is drawn through
the origin as a guide. strongly influenced by temperature both experimentally (see Fig-
ure 2) and theoretically (Figure 6), and this assignment helps
to explain the significant broadening and shifting that occurs
with increasing temperature, as the vibrational potential likely
becomes much softer and more anharmonic (leading to more re-
laxation pathways) with increasing thermal expansion (and ther-
mal energy). In order to further predict the role that thermal expansion plays
of the structures and vibrational dynamics of the PCB crystals,
QHA simulations were performed. The QHA method performs
volume-constrained geometry optimisations (all lattice vectors
and atomic positions were allowed to relax while maintaining a
constant volume) and frequency analyses at four distinct volumes
corresponding to the equilibrium (no volume constraint) struc-
ture, and −2,+2, and +4% contracted/expanded volumes. Based
on these results, the temperature dependence of the vibrational
features can be estimated within the context of thermal expan-
sion effects. Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36.
This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. The
well-resolved experimental transition at 1.23THz, and predicted
at 1.337THz, involves a coupled hindered rotation of all of the
molecules within the solid along their long-axes (see Figure 3),
which at its maximum displacement yields a totally planar ar-
rangement of all of the molecules within the solid. Thus, this
mode corresponds well to the phase-transformation pathway pre-
dicted from the AIMD simulations. Interestingly, this mode is In both simulations, the individual molecules undergo predom-
inate rocking motions with respect to their neighbours, which is
evident in the periodic fluctuations present in Figure 3. Even in Faraday Discussions Faraday Discussions Page 5 of 9 Page 5 of 9 Fig. 3 Bulk static structures of the two forms of PCB, the low-temperature ordered α-form (left) and the high-temperature plastic γ-crystal (right). The
main structural difference between the two crystals is highlighted in orange, along with arrows denoting the hindered-rotational motion that drives the
transformation between the two forms and corresponds to the vibrations discussed later in the text. View Article Online
DOI: 10.1039/C8FD00042E View Article Online
DOI: 10.1039/C8FD00042E Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. scussions Accepted Manuscript s Accepted Manuscript Faraday Discussions Accepted Manuscript Fig. 3 Bulk static structures of the two forms of PCB, the low-temperature
main structural difference between the two crystals is highlighted in orang
transformation between the two forms and corresponds to the vibrations di
Fig. 4 AIMD simulated angles between adjacent molecular planes at
100K (blue) and 300K (red). A horizontal dashed line is drawn through
the origin as a guide. Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. Fig. 3 Bulk static structures of the two forms of PCB, the low-temperature ordered α-form (left) and the high-temperature plastic γ-crystal (right). The
main structural difference between the two crystals is highlighted in orange, along with arrows denoting the hindered-rotational motion that drives the
transformation between the two forms and corresponds to the vibrations discussed later in the text. araday Discussions Accepted Man lations between pairs of neighbouring molecules (see Figure 4),
and also corresponds well to the oscillation period, indicating that
this particular feature is indeed the local hindered-rotational mo-
tion suggested. Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36.
This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. 5 Experimental THz-TDS spectra (blue) and s-DFT simulated IR-spectra
have a number of IR-active modes which primarily correspond to hindered-rota
contains fewer IR-active modes (due to the smaller static unit cell compared t
lines) predicted that might be activated in the experimental disordered structure
red lines, and are in good general agreement with the experimental spectra. Fig. 6 AIMD-simulated IR-spectra of α-PCB at 100,200, and 250K (blue,
purple, and red, respectively). The spectra exhibit a significant red-shift
and spectral broadening as the temperature is increased. co
ro
fo
h
u
in
3
T
si
p
te
D
a
n
o
m
a
cu
su
W
tr
m
ti
e
γ
3
(F
Open Access Article. Published on 16 April 2018. Downloaded on 04/0
This article is licensed under a Creative Commons Attribut Fig. 5 Experimental THz-TDS spectra (blue) and s-DFT simulated IR-spectra (black) for both PCB solids. The α-PCB spectrum (80K) is predicted to
have a number of IR-active modes which primarily correspond to hindered-rotational motions of the individual molecules within the solid. The γ-form
contains fewer IR-active modes (due to the smaller static unit cell compared to the α-form), however there are additional IR-inactive modes (dotted
lines) predicted that might be activated in the experimental disordered structure. In both panels, the simulated AIMD spectra are also shown as dotted
red lines, and are in good general agreement with the experimental spectra. corresponding to the phase-transformation mode (i.e. hindered
rotations about the molecular plane) involve even larger shifts,
for example the mode at 1.337THz in the α-form predicted to
have a 26cm−1 red-shift. These data highlight the importance of
unit cell dimensions on the low-frequency vibrational potentials
in molecular solids. Fig. 6 AIMD-simulated IR-spectra of α-PCB at 100,200, and 250K (blue,
purple, and red, respectively). The spectra exhibit a significant red-shift
and spectral broadening as the temperature is increased. Faraday Discussions Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36.
This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. The results indicate that all of the low-frequency
vibrations in both crystals are dependent on the volume to a
large extent, with most modes experiencing shifts on the order of
10cm−1 between the two extreme volumes. This is also reflected
in the temperature-dependent AIMD simulations (see Figure 6),
which also predicts significant red-shifting upon increasing tem-
perature. Remarkably, the vibrational modes that involve motions In the γ-form, there are fewer IR-active modes predicted due
to fewer molecules within the crystallographic unit cell. The re-
solvable features at 1.72THz can be assigned to an intense mode
predicted at 1.755THz. This is a surprising result given most dis-
ordered materials do not contain discernible low-frequency ab-
sorption features, and it is even more surprising that the static-
model is able to reproduce the disordered experimental results. Investigation of the vibrational mode type highlights that the vi-
brational motion is an asymmetric rocking of two molecules about
their long axes with respect to one another. This directly cor-
responds to the coupled oscillations present in the AIMD simu- Journal Name, [year], [vol.],
1–9 | 5 Faraday Discussions Faraday Discussions Page 6 of 9 Fig. 5 Experimental THz-TDS spectra (blue) and s-DFT simulated IR-spectra (black) for both PCB solids. The α-PCB spectrum (80K) is predicted to
have a number of IR-active modes which primarily correspond to hindered-rotational motions of the individual molecules within the solid. The γ-form
contains fewer IR-active modes (due to the smaller static unit cell compared to the α-form), however there are additional IR-inactive modes (dotted
nes) predicted that might be activated in the experimental disordered structure. In both panels, the simulated AIMD spectra are also shown as dotted
ed lines, and are in good general agreement with the experimental spectra. View Article Online
DOI: 10.1039/C8FD00042E View Article Online
DOI: 10.1039/C8FD00042E Fig. 5 Experimental THz-TDS spectra (blue) and s-DFT simulated IR-spectra (black) for both PCB solids. The α-PCB spectrum (8
have a number of IR-active modes which primarily correspond to hindered-rotational motions of the individual molecules within the
contains fewer IR-active modes (due to the smaller static unit cell compared to the α-form), however there are additional IR-inac
lines) predicted that might be activated in the experimental disordered structure. In both panels, the simulated AIMD spectra are als
red lines, and are in good general agreement with the experimental spectra. Fig. 6 |
1–9
Journal Name, [year], [vol.], Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36.
This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. 6 AIMD-simulated IR-spectra of α-PCB at 100,200, and 250K (blue,
purple, and red, respectively). The spectra exhibit a significant red-shift
and spectral broadening as the temperature is increased. corresponding to the phase-transformation mod
rotations about the molecular plane) involve ev
for example the mode at 1.337THz in the α-fo
have a 26cm−1 red-shift. These data highlight th
unit cell dimensions on the low-frequency vibra
in molecular solids. 3.4
Thermodynamic Analysis
The accurate modelling of the vibrational spectr
simulations lends confidence to the theoretical m
ployed, and enables the prediction of thermod
ters to be performed. It is important to note t
DFT predicted-IR (and Raman) activities might n
accurate for certain modes in the disordered-cr
namic functions depend more simply on the corr
of the vibrational states, regardless of their sym
monic canonical vibrational partition function
and related quantities such as entropy and free
culated. All energies discussed have been corre
superposition error using the counterpoise corre
When strictly considering the electronic energy,
trinsic temperature or pressure dependence, the
more stable system by 0.56kJ mol−1. However,
tional results are used to determine the temper
entropic and free energy parameters, the results
γ-solid becomes the more thermodynamically sta
301K, close to the experimental transition temp
(Figure 7). In an attempt to increase the accuracy of the ca
Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. Fig. 5 Experimental THz-TDS spectra (blue) and s-DFT simulated IR-spe
have a number of IR-active modes which primarily correspond to hindered
contains fewer IR-active modes (due to the smaller static unit cell compa
lines) predicted that might be activated in the experimental disordered stru
red lines, and are in good general agreement with the experimental spectr
Fig. 6 AIMD-simulated IR-spectra of α-PCB at 100,200, and 250K (blue,
purple, and red, respectively). The spectra exhibit a significant red-shift
and spectral broadening as the temperature is increased. Open Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. ssions Accepted Manuscript Accepted Manuscript y Discussions Accepted Manuscrip Fig. 3.4
Thermodynamic Analysis The accurate modelling of the vibrational spectrum by the s-DFT
simulations lends confidence to the theoretical methodology em-
ployed, and enables the prediction of thermodynamic parame-
ters to be performed. It is important to note that while the s-
DFT predicted-IR (and Raman) activities might not be completely
accurate for certain modes in the disordered-crystal, thermody-
namic functions depend more simply on the correct identification
of the vibrational states, regardless of their symmetry. The har-
monic canonical vibrational partition function was determined,
and related quantities such as entropy and free energy were cal-
culated. All energies discussed have been corrected for basis set
superposition error using the counterpoise correction method.58
When strictly considering the electronic energy, which has no in-
trinsic temperature or pressure dependence, the α-crystal is the
more stable system by 0.56kJ mol−1. However, when the vibra-
tional results are used to determine the temperature-dependent
entropic and free energy parameters, the results suggest that the
γ-solid becomes the more thermodynamically stable crystal above
301K, close to the experimental transition temperature of 317K
(Figure 7). Fig. 6 AIMD-simulated IR-spectra of α-PCB at 100,200, and 250K (blue,
purple, and red, respectively). The spectra exhibit a significant red-shift
and spectral broadening as the temperature is increased. In an attempt to increase the accuracy of the calculated thermo-
dynamic constants, the QHA vibrational results were used to de-
termine the temperature-dependent free energies. Because these
simulations explicitly take thermal expansion into account, they 6 |
1–9
Journal Name, [year], [vol.], Faraday Discussions Page 7 of 9 Page 7 of 9 View Article Online Fig. 7 Harmonic (top) and quasi-harmoinc (bottom) free energy curves
for the α- and γ-PCB crystals (blue and red, respectively). The verti-
cal dotted line is drawn at the intersection of the two curves, occurring
at 301K and 326K for the harmonic and quasi-harmonic simulations, re-
spectively. In both charts the data have been offset relative to the value
of the α-form free energy at 50K. Creative Commons Attribution 3.0 Unported Licence. the forces, including the weak intermolecular interactions, pro-
viding a high-level of confidence in them. However, not all sys-
tems are well suited for study using this methodology, particularly
in predicting large-scale structural reorganisations, which would
require much longer AIMD simulations in order to capture the ef-
fect. Acknowledgements The authors thank the UK Engineering and Physical Sciences Re-
search Council (EPSRC) for funding (EP/N022769/1). Simula-
tions were possible via our membership of the UK’s HEC Ma-
terials Chemistry Consortium, which is funded by the EPSRC
(EP/L000202), and this work used the ARCHER UK National
Supercomputing Service (http://www.archer.ac.uk). JAZ and JK
thank the EPSRC Cambridge Centre for Doctoral Training in Sen-
sor Technologies and Applications (EP/L015889/1). 3.5
Interpretation and Further Developments of Proposed
Method The combination of the experimental and theoretical results en-
ables a great deal of information surrounding the phase transi-
tion behaviour in molecular solids to be understood. Thus, such a
methodology is generally applicable to a large variety of materi-
als, particularly those that undergo diffusionless transformations. The sensitivity of THz-TDS measurements to the bulk packing ar-
rangement, any minor deviations from ideal crystalline ordering,
leads to it being a powerful probe of the processes and dynamics
occurring within solid materials. While X-ray diffraction is often
used for interpreting this type of behaviour, it is in reality a com-
plement to the THz-TDS experiment, as the former provides in-
formation related to the average structure, while the latter probes
molecular dynamics. The addition of computational results pro-
vides a means of interpreting the experimental spectra, indicat-
ing what types of motions are occurring within the solids. Addi-
tionally, the accurate reproduction of the experimental THz-TDS
spectra indicates that the simulations are correctly reproducing 3.4
Thermodynamic Analysis Nonetheless, the proposed methodology is well suited for
order-disorder transitions, particularly in molecular solids where
such transformations occur near ambient conditions. DOI: 10.1039/C8FD00042E ssions Accepted Manuscript 4
Conclusions Accepted Manuscript Faraday Discussions Accepted Manuscript Overall, the proposed combination of experimental terahertz
time-domain spectroscopy and theoretical ab initio molecular dy-
namics and static-solid-state-density functional theory methods
provides a complete means to elucidate the structures, phase tran-
sition mechanisms, and bulk properties (e.g. thermodynamics and
mechanochemical effects such as thermal expansion, and other
related quantities) of molecular solids. The results shed light on
the importance of low-frequency vibrational modes on many as-
pects of the properties of disordered crystals, and the atomic-level
details provided by the s-DFT simulations enable experimental
observations regarding temperature-dependent spectral trends to
be fully rationalised. And while traditional harmonic approxima-
tion vibrational analyses were shown to be a suitable means of
determining the thermodynamic properties of molecular crystals,
the results can be significantly improved by taking into account
thermal expansion effects via the use of quasi-harmonic approx-
imation simulations, which provide a more physical (and accu-
rate) description of both the temperature-dependent vibrational
and thermodynamical forces present within such materials. Discussions Accepted Manuscri Fig. 7 Harmonic (top) and quasi-harmoinc (bottom) free energy curves
for the α- and γ-PCB crystals (blue and red, respectively). The verti-
cal dotted line is drawn at the intersection of the two curves, occurring
at 301K and 326K for the harmonic and quasi-harmonic simulations, re-
spectively. In both charts the data have been offset relative to the value
of the α-form free energy at 50K. are able to provide a more physical description of the thermody-
namic forces involved in these materials. The simulated QHA free
energy curves, also shown in Figure 7, not only show a larger
deviation in energy between the two solids, but also provide a
more accurate description of the phase-transition temperature,
predicted at 326K, only 9K higher than the experimentally ob-
served temperature. The greater accuracy achieved when explic-
itly considering the thermal expansion properties of the solids,
while not surprising, highlights the importance of taking into ac-
count these forces when attempting to describe the thermody-
namics of weakly bound molecular crystals. Faraday Discussions n Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36.
This article is licensed under a Creative Commons Attribution 3.0 Unported Licenc 38 G. J. Martyna, D. J. Tobias and M. L. Klein, J. Chem. Phys.,
1994, 101, 4177–4189. 11 M. D. King, T. N. Blanton, S. T. Misture and T. M. Korter,
Cryst. Growth Des., 2011, 11, 5733–5740. Discussions Accepted Manuscri 39 G. J. Martyna, M. E. Tuckerman, D. J. Tobias and M. L. Klein,
Mol. Phys., 1996, 87, 1117–1157. 12 M. T. Ruggiero, J. Sibik, R. Orlando, J. A. Zeitler and T. M. Korter, Angew. Chem. Int. Ed., 2016, 55, 6877–6881. 40 J. VandeVondele and J. Hutter, J. Chem. Phys., 2007, 127,
114105. 13 J. Timmermans, J. Phys. Chem. Solids, 1961, 18, 1–8. 41 S. Goedecker and M. Teter, Phys. Rev. B, 1996, 54, 1703–
1710. 14 J. M. Pringle, P. C. Howlett, D. R. MacFarlane and M. Forsyth,
J. Mater. Chem., 2010, 20, 2056–2062. 42 J. Perdew, K. Burke and M. Ernzerhof, Phys. Rev. Lett., 1996,
77, 3865–3868. 15 M. Descamps, N. T. Correia, P. Derollez, F. Danede and
F. Capet, J. Phys. Chem. B, 2005, 109, 16092–16098. 16 F. Affouard, J. F. Willart and M. Descamps, author, 2002, pp. 9–15. 43 S. Grimme, J. Antony, S. Ehrlich and H. Krieg, J. Chem. Phys.,
2010, 132, 154104. 44 S. Grimme, S. Ehrlich and L. Goerigk, J. Comput. Chem., 2011,
32, 1456–1465. 17 A. Hédoux, Y. Guinet, P. Derollez, J.-F. Willart, F. Capet and
M. Descamps, J. Phys.: Condens. Matter, 2003, 15, 8647–
8661. 45 M. Brehm and B. Kirchner, J. Chem. Inf. Model., 2011, 51,
2007–2023. 18 L. Jin, K. M. Nairn, C. M. Forsyth, A. J. Seeber, D. R. MacFar-
lane, P. C. Howlett, M. Forsyth and J. M. Pringle, J. Am. Chem. Soc., 2012, 134, 9688–9697. 46 R. Dovesi, R. Orlando, A. Erba, C. M. Zicovich-Wilson, 46 R. Dovesi, R. Orlando, A. Erba, C. M. Zicovich-Wilson,
B. Civalleri,
S. Casassa,
L. Maschio,
M. Ferrabone,
M. De La Pierre, P. D’Arco, Y. Noël, M. Causà, M. Rérat and
B. Kirtman, Int. J. Quantum Chem., 2014, 114, 1287. Faraday Discussions A 19 R. Brand, P. Lunkenheimer and A. Loidl, J. Chem. Phys., 2002,
116, 10386–10401. Discussions 20 F. Affouard and M. Descamps, Phys. Rev. Lett., 2001, 87,
035501. 47 F. Weigend and R. Ahlrichs, Phys. Chem. Chem. Phys., 2005,
7, 3297–3305. 48 F. Pascale, C. M. Zicovich-Wilson, F. L. Gejo, B. Civalleri,
R. Orlando and R. Dovesi, J. Comput. Chem., 2004, 25, 888–
897. 21 F. Affouard, E. Notes and references 1 Veaceslav Coropceanu, Jérôme Cornil, Demetrio A da Silva
Filho, Yoann Olivier, R. Silbey, and Jean-Luc Brédas, Chem. Rev., 2007, 107, 926–952. 1 Veaceslav Coropceanu, Jérôme Cornil, Demetrio A da Silva
Filho, Yoann Olivier, R. Silbey, and Jean-Luc Brédas, Chem. Rev., 2007, 107, 926–952. 2 S. B. Murdande, M. J. Pikal, R. M. Shanker and R. H. Bogner,
J. Pharm. Sci., 2010, 99, 1254–1264. 3 A. Cavagna, Phys. Rep., 2009, 476, 51–124. 3 A. Cavagna, Phys. Rep., 2009, 476, 51–124. 4 M. Oguni and N. Okamotot, Solid State Commun., 1996, 99,
53–56. 4 M. Oguni and N. Okamotot, Solid State Commun., 1996, 99,
53–56. 5 M. T. Ruggiero, M. Krynski, E. O. Kissi, J. Sibik, D. Markl, N. Y. Tan, D. Arslanov, W. van der Zande, B. Redlich, T. M. Korter,
H. Grohganz, K. Löbmann, T. Rades, S. R. Elliott and J. A. Zeitler, Phys. Chem. Chem. Phys., 2017, 19, 30039–30047. Journal Name, [year], [vol.],
1–9 | 7 Faraday Discussions Page 8 of 9 Page 8 of 9 View Article Online 6 E. Shalaev and A. K. Soper, J. Phys. Chem. B, 2016, 120, 7289–
7296. 33 Y. Takaki, K. Nakata, T. Taniguchi and K. Sakurai, Acta Crys-
tallogr. B, 1978, 34, 2579–2586. DOI: 10.1039/C8FD00042E 7 M. T. Ruggiero, J. Axel Zeitler and T. M. Korter, Phys. Chem. Chem. Phys., 2017, 19, 28502–28506. 34 J. Hutter, M. Iannuzzi, F. Schiffmann and J. VandeVondele,
WIREs Comput Mol Sci, 2013, 4, 15–25. 8 E. P. J. Parrott and J. A. Zeitler, Appl. Spectrosc., 2015, 69,
1–25. 35 J. VandeVondele, M. Krack, F. Mohamed, M. Parrinello,
T. Chassaing and J. Hutter, Comput. Phys. Commun., 2005,
167, 103–128. 9 J. A. Zeitler, P. F. Taday, D. A. Newnham, M. Pepper, K. C. Gordon and T. Rades, J. Pharm. Pharmacol., 2007, 59, 209–
223. 36 S. Nosé, J. Chem. Phys., 1998, 81, 511–519. 37 S. Nosé, Mol. Phys., 2002, 100, 191–198. ns Accepted Manuscript Faraday Discussions Accepted Manuscript 10 G. J. O. Beran, Chem. Rev., 2016, 116, 5567–5613. 58 M. Gutowski and G. Chalasi´nski, J. Chem. Phys., 1998, 98,
5540–5554.
View Article Online
DOI: 10.1039/C8FD00042E Appl. Phys. Lett., 2003, 82, 2350–2352.
58 M. Gutowski and G. Chalasi´nski, J. Chem. Phys., 1998, 98,
5540–5554.
9 of 9
Faraday Discussions
View Article Online
DOI: 10.1039/C8FD00042E Page 9 of 9 Page 9 of 9 n Access Article. Published on 16 April 2018. Downloaded on 04/06/2018 13:47:36.
This article is licensed under a Creative Commons Attribution 3.0 Unported Licenc Cochin, R. Decressain and M. Descamps, Euro-
phys. Lett., 2001, 53, 611–617. 22 O. B. M. H. Duparc and M. Meyer, J. Chem. Phys., 1998, 93,
1313–1324. 49 C. M. Zicovich-Wilson, F. Pascale, C. Roetti, V. R. Saunders,
R. Orlando and R. Dovesi, J. Comput. Chem., 2004, 25, 1873–
1881. 23 J. Adebahr, F. C. Grozema, S. W. deLeeuw, D. R. MacFarlane
and M. Forsyth, Solid State Ion., 2006, 177, 2845–2850. 50 Y. Noël, C. Zicovich-Wilson, C. M. Zicovich-Wilson, R. Dovesi,
B. Civalleri, P. D’Arco and R. Dovesi, Phys. Rev. B, 2001, 65,
014111–9. 24 M. Thomas, M. Brehm, R. Fligg, P. Vöhringer and B. Kirchner,
Phys. Chem. Chem. Phys., 2013, 15, 6608–6622. Faraday 25 M. Yamaguchi and A. Ohira, Comput. Theor. Chem., 2016,
1089, 54–58. 51 A. Erba, J. Maul and B. Civalleri, Chem. Commun., 2016, 52,
1820–1823. 26 M. Thomas, M. Brehm and B. Kirchner, Phys. Chem. Chem. Phys., 2015, 17, 3207–3213. 52 A. Erba, M. Shahrokhi, R. Moradian and R. Dovesi, J. Chem. Phys., 2015, 142, 044114. 27 M. T. Ruggiero and J. A. Zeitler, J. Phys. Chem. B, 2016, 120,
11733–11739. 53 M. T. Ruggiero, J. A. Zeitler and A. Erba, Chem. Commun.,
2017, 53, 3781–3784. 28 G. Kresse and J. Hafner, Phys. Rev. B, 1994, 49, 14251–14269. 54 J. G. Brandenburg, J. Potticary, H. A. Sparkes, S. L. Price and
S. R. Hall, J. Phys. Chem. Lett., 2017, 8, 4319–4324. 29 A. S. Larsen, M. T. Ruggiero, K. E. Johansson, J. A. Zeitler and
J. Rantanen, Cryst. Growth Des., 2017, 17, 5017–5022. 55 M. T. Ruggiero, T. Bardon, M. Strliˇc, P. F. Taday and T. M. Korter, Phys. Chem. Chem. Phys., 2015, 17, 9326–9334. 30 S. Tsuneyuki, H. Aoki, M. Tsukada and Y. Matsui, Phys. Rev. Lett., 1990, 64, 776–779. 56 P. G. Karamertzanis and C. C. Pantelides, Mol. Phys., 2007,
105, 273–291. 31 A. Kubo, R. Ikeda and D. Nakamura, Z. Naturforsch. A, 1986,
41, 270–274. 57 Y. C. Shen, P. C. Upadhya, E. H. Linfield and A. G. Davies, 32 A. Schönleber, P. Pattison and G. Chapuis, Z. Kristallogr. Cryst. Mater., 2003, 218, 47. 8 |
1–9
Journal Name, [year], [vol.], Journal Name, [year], [vol.],
1–9 | 9 Appl. Phys. Lett., 2003, 82, 2350–2352. Faraday Discussions Appl. Phys. Lett., 2003, 82, 2350–2352. ssions Accepted Manuscript Faraday Discussions Accepted Manuscript y Discussions Accepted Manuscrip Faraday Discussions Accepted Man Faraday Discussions Acce Journal Name, [year], [vol.],
1–9 | 9
|
https://openalex.org/W2950679151
|
https://www.research.ed.ac.uk/files/122304568/Gradual_session_types_IGARASHI_DOA25112019_VOR_CC_BY.pdf
|
English
| null |
Gradual session types
|
Journal of functional programming
| 2,019
|
cc-by
| 32,507
|
Edinburgh Research Explorer General rights
C
i h f
h General rights
Copyright for the publications made accessible via the Edinburgh Research Explorer is retained by the author(s)
and / or other copyright owners and it is a condition of accessing these publications that users recognise and
abide by the legal requirements associated with these rights. Digital Object Identifier (DOI):
10.1017/S0956796819000169 Document Version:
Publisher's PDF, also known as Version of record Published In:
Journal of Functional Programming Gradual session types Citation for published version:
Igarashi, A, Thiemann, P, Tsuda, Y, Vasconcelos, VT & Wadler, P 2019, 'Gradual session types', Journal of
Functional Programming, vol. 29, e17. https://doi.org/10.1017/S0956796819000169 Citation for published version:
Igarashi, A, Thiemann, P, Tsuda, Y, Vasconcelos, VT & Wadler, P 2019, 'Gradual session types', Journal of
Functional Programming, vol. 29, e17. https://doi.org/10.1017/S0956796819000169 Citation for published version: Abstract Session types are a rich type discipline, based on linear types, that lifts the sort of safety claims
that come with type systems to communications. However, web-based applications and microser-
vices are often written in a mix of languages, with type disciplines in a spectrum between static and
dynamic typing. Gradual session types address this mixed setting by providing a framework which
grants seamless transition between statically typed handling of sessions and any required degree of
dynamic typing. We propose Gradual GV as a gradually typed extension of the functional session
type system GV. Following a standard framework of gradual typing, Gradual GV consists of an
external language, which relaxes the type system of GV using dynamic types; an internal language
with casts, for which operational semantics is given; and a cast-insertion translation from the former
to the latter. We demonstrate type and communication safety as well as blame safety, thus extend-
ing previous results to functional languages with session-based communication. The interplay of
linearity and dynamic types requires a novel approach to specifying the dynamics of the language. Take down policy Take down policy
The University of Edinburgh has made every reasonable effort to ensure that Edinburgh Research Explorer
content complies with UK legislation. If you believe that the public display of this file breaches copyright please
contact openaccess@ed.ac.uk providing details, and we will remove access to the work immediately and
investigate your claim. Download date: 24. Oct. 2024 1
s JFP 29, e17, 56 pages, 2019. c⃝The Author(s) 2019. This is an Open Access article, distributed under
1
the terms of the Creative Commons Attribution licence (http://creativecommons.org/licenses/by/4.0/), which permits
unrestricted re-use, distribution, and reproduction in any medium, provided the original work is properly cited. doi:10.1017/S0956796819000169 JFP 29, e17, 56 pages, 2019. c⃝The Author(s) 2019. This is an Open Access article, distributed under
1
the terms of the Creative Commons Attribution licence (http://creativecommons.org/licenses/by/4.0/), which permits
unrestricted re-use, distribution, and reproduction in any medium, provided the original work is properly cited. doi:10.1017/S0956796819000169 1 https://www.ubuntupit.com/top-20-most-popular-programming-languages-to-learn-for
your-open-source-project/ accessed in April 2019. your-open-source-project/ accessed in April 2019. ttps://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
ownloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 1 Introduction It was the best of types, it was the worst of types. A survey of the top-20 programming languages to learn for open source projects1 lists
eight dynamically typed languages (JavaScript, Python, Ruby, R, PHP, Perl, Scheme, and https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at A. Igarashi et al. 2 Erlang) and states that developers’ salaries for these languages are among the highest in the
industry. The survey also suggests to learn languages with elaborate static type systems like
Rust, Scala, and Haskell, with developers earning even higher salaries. These languages
derive their expressiveness from advanced type system features like linearity, uniqueness,
effects, and dependent types as embodied in research languages like Agda (Norell, 2009),
Coq (The Coq Development Team, 2019), and Idris (Brady, 2013); and session types as
in Links (Lindley & Morris, 2016b). These data indicate two opposing trends in current
industrial practice, one asking for dynamically typed programming and another asking for
expressive statically typed programming. Gradually typed languages reconcile these two trends. They permit one to assemble
programs with some components written in a statically typed language and some in a
dynamically typed language. Gradually typed languages have been widely explored in
both theory and practice, beginning with contracts in Racket (Findler & Felleisen, 2002)
and their interfacing with TypedRacket (Tobin-Hochstadt & Felleisen, 2008) and then
popularized by Siek and others (Siek & Taha, 2006, 2007; Siek et al., 2015b). They are
geared toward safely interconnecting dynamically typed parts with statically typed parts
of a program by ensuring that type mismatches only occur in the dynamically typed parts
(Wadler & Findler, 2009). Dynamics in C# (Bierman et al., 2010), Microsoft’s TypeScript2 (Bierman et al., 2014),
Google’s Dart (The Dart Team, 2014; Ernst et al., 2017), and Facebook’s Hack (Verlaguet,
2013) and Flow (Chaudhuri et al., 2017) are industrial systems inspired by gradual typing,
but focusing on enhancing programmer productivity and bug finding rather than contain-
ing type mismatches. Systems such as Racket (Findler & Felleisen, 2002) and Reticulated
Python (Vitousek et al., 2017) rely on contracts or similar constructs to ensure that dynam-
ically typed values adhere to statically typed constraints when values pass from one world
to the other. 2 https://www.typescriptlang.org/ accessed in April 2019. 3 http://estafet.com/scribble/ Accessed in April 2019. 1 Introduction At first blush, one might consider gradual types as largely a response to the former
trend: they provide a way for developers using dynamically typed languages to evolve
their code toward statically typed languages that are deemed easier to maintain. But on
second thought, one might consider gradual types as even more helpful in light of the
latter trend. Suitably generalized, gradual typing can mediate between simple type systems
and type systems that feature dependent types, effect types, or session types, for example. Gradual typing in this sense can help in evolving software development toward languages
with more precise type systems. Hence, an important line of research is to extend gradual typing so that it not only relates
dynamically typed and statically typed languages, but also relates less-precisely typed and
more-precisely typed languages. There is already some research on doing so for dependent
types (Ou et al., 2004; Flanagan, 2006; Greenberg et al., 2010; Lehmann & Tanter, 2017),
effect types (Bañados Schwerter et al., 2014), typestate (Wolff et al., 2011), and several
others which we review in the section on related work. This paper presents the first system
that extends gradual typing to session types. Session types were introduced by Honda (1993), drawing on Milner’s π-calculus
(Milner et al., 1992) and Girard’s linear logic (1987), and further developed by many others 3 Gradual session types (Honda et al., 1998; Yoshida & Vasconcelos, 2007). Gay and Hole (2005) introduced sub-
typing for session types, and session types were embedded into a functional language with
linear types, similar to the one used in this paper, by Gay and Vasconcelos (2010). Caires,
Pfenning, Toninho, and Wadler introduced propositions-as-types interpretations of session
types in linear logic (Caires & Pfenning, 2010; Wadler, 2012, 2014; Caires et al., 2014). One important line of research is multiparty session types (Honda et al., 2008, 2016) but
we confine our attention here to dyadic session types. (Honda et al., 1998; Yoshida & Vasconcelos, 2007). Gay and Hole (2005) introduced sub-
typing for session types, and session types were embedded into a functional language with
linear types, similar to the one used in this paper, by Gay and Vasconcelos (2010). Caires,
Pfenning, Toninho, and Wadler introduced propositions-as-types interpretations of session
types in linear logic (Caires & Pfenning, 2010; Wadler, 2012, 2014; Caires et al., 2014). 1 Introduction One important line of research is multiparty session types (Honda et al., 2008, 2016) but
we confine our attention here to dyadic session types. Session types have been adapted to a variety of languages, either statically or dynam-
ically checked, and using either libraries or additions to the toolchain; implementa-
tions include C, Erlang, Go, Haskell, Java, Python, Rust, and Scala. New languages
incorporating session types include C0 (Willsey et al., 2017), Links (Cooper et al.,
2007), SePi (Franco & Vasconcelos, 2013), SILL (Pfenning & Griffith, 2015), and
Singularity (Fähndrich et al., 2006). Industrial uses of session types include Red Hat’s
support of the Scribble specification language (Yoshida et al., 2014), which has been used
as a common interface for several systems based on session types; Estafet’s use of session
types to manage microservices3; and the Ocean Observatories Initiative’s use of dynami-
cally checked session types in Python (Demangeon et al., 2015). Session types inspired an
entire line of research on what has come to be called behavioral types, the subject of EU
COST action BETTY, a recent Shonan meeting, and a recent Dagstuhl seminar. Here is a simple session type encoding of a protocol to purchase an online video: Svideo = !string. ?int. ⊕{buy : !CC. ?URL. end?, quit : end!} It describes a channel endpoint along which a client sends the name of a video as a string,
receives its cost as an integer, and then selects either to buy the video, in which case one
sends a credit card number, receives a URL from which the video may be downloaded,
and waits for an indication that the channel has been closed, or selects to quit and closes
the channel. There is a dual session type for server at the other end of the channel, where
! (write) is swapped with ? (read), ⊕(select from a choice) is swapped with & (offer a
choice), and end! (close a channel) is swapped with end? (wait for a channel to close). Session types are necessarily linear. Let x be bound to a string and let c be bound to a
channel endpoint of type Svideo. Performing let d = send x c in . . . https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at let d = send x c in . . . Gommerstadt and
others (2016) consider dynamic monitoring of higher-order session-typed processes in the
presence of unreliable communication and malicious communication partners. Their focus
is on assigning blame correctly in this setting. The same authors (Gommerstadt et al., 2018)
develop a theory of contracts that translate into processes that serve as proxies between the
original communication partners. Proxies ensure adherence to the session protocol with
dynamic tests. A similar proxy-based monitoring scheme was also proposed by one of the
authors (Thiemann, 2014) where gradual typing was restricted to the transmitted values. Melgratti & Padovani (2017) propose a contract system that mediates between (simply
typed) sessions and contract-refined sessions. Enforcement is done with an inline monitor. In contrast to these approaches, our work applies to the mediation between dynamically
typed and statically typed code and it relies on gradual principles that enable a pay-as-you-
go approach: a protocol is checked statically as much as possible, dynamic checks are only
employed if they cannot be avoided. Our work is the first to enable full gradualization that
includes the typing of the communication channel rather than just the values transmitted. Moreover, we improve the efficiency over prior approaches by avoiding the introduction
of extra proxy processes. We give our system a compact formulation along the lines of the blame calculus
(Wadler & Findler, 2009), based on the notion of a cast to mediate interactions between
more-precisely typed (e.g., statically typed) and less-precisely typed (e.g., dynamically
typed) components of a program. We define the four subtyping relations exhibited by
the blame calculus, ordinary, positive, negative, and naive, and show the corresponding
results, including a tangram theorem relating the four forms of subtyping and blame safety. A corollary of our results is that in any interaction between more-precisely typed and less-
precisely typed components of a program, any cast error is due to the less-precisely typed
component. Our paper makes the following contributions: Our paper makes the following contributions: • Section 2 provides an overview of the novel techniques in our work, and how we
dynamically enforce linearity and session types. • Section 2 provides an overview of the novel techniques in our work, and how we
dynamically enforce linearity and session types. let d = send x c in . . . binds d to a channel endpoint of type R, where Svideo = !string.R. To avoid sending a string
to the same channel twice, it is essential that c must be bound to the only reference to
the channel endpoint before the operation, and for similar reasons d must be bound to the
only reference to the channel endpoint after. Such restrictions can easily be enforced in a
statically typed language with an affine type discipline. Linearity is required to guarantee
that channels are not abandoned before they are closed. binds d to a channel endpoint of type R, where Svideo = !string.R. To avoid sending a string
to the same channel twice, it is essential that c must be bound to the only reference to
the channel endpoint before the operation, and for similar reasons d must be bound to the
only reference to the channel endpoint after. Such restrictions can easily be enforced in a
statically typed language with an affine type discipline. Linearity is required to guarantee
that channels are not abandoned before they are closed. But how is one to ensure linearity in a dynamically typed language? Following Tov &
Pucella (2010), we require that each dynamically typed reference to a channel endpoint is
equipped with a lock. That reference is locked after the channel is used once to ensure it
cannot be used again. To ensure that each channel is appropriately terminated, with either a 4 A. Igarashi et al. wait or a close operation, garbage collection flags an error if a dynamically typed reference
to a channel becomes inaccessible. Our system is the first to integrate static and dynamic session types via gradual typing. It preserves the safety properties of statically typed sessions, namely progress (for expres-
sions), preservation, and absence of runtime errors. The latter includes session fidelity:
every send is matched with a receive, every select is matched with an offer, and every
wait is matched with close. Many, but not all, systems with session types support recursive
session types, and many, but not all, systems with session types ensure deadlock freedom;
we leave such developments for future work. Previous systems that perform dynamic monitoring on session types include the work on
Scribble (Yoshida et al., 2014) which applies the ideas developed for distributed monitor-
ing of protocols to multiparty session types (Bocchi et al., 2013, 2017). https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Camb 4 GV is our name for the functional session type calculus of Gay and Vasconcelos (2010), which is the statically
typed baseline for our gradual system. 2 Motivation Sy and Rob collaborate on a project whose design is based on microservices. Sy is a strong
advocate of static typing and relies on an implementation language that supports session
types out of the box. Rob, on the other hand, is a strong advocate of dynamically typed
languages. One of the credos of microservice architectures is that the implementation of
a service endpoint is language-agnostic, which means it can be implemented in any pro-
gramming language whatsoever as long as it adheres to its protocol. However, Sy does not
want to compromise the strong guarantees (e.g., type safety, session fidelity) of the stati-
cally typed code by communicating with Rob’s client. Rob is also keen on having strong
guarantees, but does not mind if they are enforced at runtime. Here is the story how they
can collaborate safely using Gradual GV,4 our proposal for a gradually typed functional
language with synchronous binary session types. let d = send x c in . . . • Section 3 describes a complete formal calculus, including syntax of both an exter-
nal language, in which programs are written and runtime checking is implicit, and
an internal language, in which programs are executed after runtime checking in the
form of casts is made explicit; typing rules of the two languages; reduction rules
for the internal language; cast-insertion translation from the external to the internal ms. https://doi.org/10.1017/S0956796819000169
idge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 5 5 Gradual session types language; and embedding of a dynamically typed language with channel-based
communication into our calculus. language; and embedding of a dynamically typed language with channel-based
communication into our calculus. • Section 4 presents standard results for our calculus, including progress (for expres-
sions), preservation, session fidelity, the tangram theorem, blame safety, conserva-
tivity of the external language typing over fully static typing, and type preservation
of the cast insertion translation. We also discuss the gradual guarantee prop-
erty for the external language. It turns out that it fails to hold—we will analyze
counterexamples and discuss why. • Section 5 describes related work and Section 6 concludes. Compared to the previous paper (Igarashi et al., 2017a), we extend the development with
the external language, the cast-insertion translation, a type-checking algorithm, proofs
of their properties, and analysis of the failure of the gradual guarantee, as well as
more detailed proofs for the earlier results. These extensions make gradual session types
accessible for the programmer, who works in the external language. 2.1 A compute service The compute service is a simplified version of one of the protocols in Sy and Rob’s project. The service involves two peers, a server and a client, connected via a communication link. The server runs a protocol that first offers a choice of two arithmetic operations, negation
or addition, then reads one or two numbers depending on the operation, outputs the result
of applying the selected operation to its operand(s), and finally closes the connection. The
client chooses an operation by sending the server a label, which is either neg or add indicat-
ing the choice of negation or addition, respectively. In session-type notation, the server’s
view of the compute protocol reads as follows: Compute = &{neg : ?int.!int.end!, add : ?int.?int.!int.end!} Compute = &{neg : ?int.!int.end!, add : ?int.?int.!int.end!} Sy chooses to implement the server in the language GV that is inspired by previous work
(Gay & Vasconcelos, 2010) and that we will describe formally in Section 3. 4 GV is our name for the functional session type calculus of Gay and Vasconcelos (2010), which is the statically
typed baseline for our gradual system. ms. https://doi.org/10.1017/S0956796819000169
idge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at A. Igarashi et al. 6 computeServer
:
Compute →
unit
computeServer
c =
case c of {
neg :
c . l e t
v1 , c = receive
c
in
l e t
c = send (−v1 )
c
in
close c ;
add :
c . l e t
v1 , c = receive
c
in
l e t
v2 , c = receive
c
in
l e t
c = send ( v1+v2 )
c
in
close c
} } The parameter c of type Compute is the server’s endpoint of the communication link to
the client (when unambiguous, we often just say endpoint or channel). The case c of ... expression receives the client’s choice on channel c in the form of a label neg or add and
branches accordingly. The notation “c. ” in each branch (re-)binds the variable c to the
channel in the state after the transmission has happened. The type of c is updated to the
session type corresponding to the respective branch in the Compute type. 2.1 A compute service The receive c
operation receives a value on channel c and returns a pair of the received value and the
depleted channel with a correspondingly depleted session type. Analogously, the send v c
operation sends value v on channel c and returns the depleted channel. The final close c
disconnects the communication link by closing the channel. 2.2 The view from the client side A client of the Compute protocol communicates on a channel with the protocol ComputeD
defined in the following. This protocol is dual to Compute: sending and receiving
operations are swapped. 2.3 A unityped server To test some new features, Rob also implements the Compute protocol, but does so in the
unityped language Uni GV, which is safe but does not impose a static typing discipline. Here is Rob’s implementation of the server. −−unityped
dynServer
c =
case c of {
neg :
c . serveOp 1 (λx.−x )
c ;
add :
c . serveOp 2 (λx . λy . x+y )
c
} −−unityped
dynServer
c =
case c of {
neg :
c . serveOp 1 (λx.−x )
c ;
add :
c . serveOp 2 (λx . λy . x+y )
c
} serveOp n op c =
i f
n==0 then
close
(send op c )
else
l e t
v , c = receive
c
in
serveOp
(n−1) ( op v )
c serveOp n op c =
i f
n==0 then
close
(send op c )
else
l e t
v , c = receive
c
in
serveOp
(n−1) ( op v )
c serveOp n op c = The main function dynServer takes a channel c on which it receives the client’s selec-
tion. It delegates to an auxiliary function serveOp that takes the arity of a function, the
function itself, and the channel end on which to receive the arguments and to send the
result. The serveOp function counts down the number of remaining function applications
in the first argument, accumulates partial function applications in the second argument, and
propagates the channel end in the third argument. It is easy to see that the dynServer function implements the Compute protocol. Rob chose
this style of implementation because it is amenable to experimentation with protocol exten-
sions: the function dynServer is trivially extensible to new operations and types by adding
new lines to the case dispatch. ComputeD = ⊕{neg : !int.?int.end?, add : !int.!int.?int.end?} A client of the compute service may always select the same operation and then proceed
linearly according the corresponding branch. Such a client can use a simpler supertype
of ComputeD with a unary internal choice. For example, a client that only ever asks for
negation can implement ComputeDneg. ComputeDneg = ⊕{neg : !int.?int.end?} ComputeDneg = ⊕{neg : !int.?int.end?} ComputeDneg = ⊕{neg : !int.?int.end?} Here is Sy’s implementation of a typed client for ComputeDneg. n e g a t i o n C l i e n t
:
i n t
→ComputeDneg →
i n t
n e g a t i o n C l i e n t
v c =
l e t
c = select
neg c
in
l e t
c = send v c
in
l e t
y , c = receive
c
in
l e t _ = wait
c
in
y n e g a t i o n C l i e n t
:
i n t
→ComputeDneg →
i n t
n e g a t i o n C l i e n t
v c =
l e t
c = select
neg c
in
l e t
c = send v c
in
l e t
y , c = receive
c
in
l e t _ = wait
c
in
y There are two new operations in the client code. The select neg c operation selects the neg
branch in the protocol by sending the neg label to the server. It returns a channel to run the 7 Gradual session types selected branch of the protocol with type !int.?int.end?. The wait c operation matches the
close operation on the server and disconnects the client. selected branch of the protocol with type !int.?int.end?. The wait c operation matches the
close operation on the server and disconnects the client. 2.4 The gradual way How can we embed Rob’s server with other program fragments in the typed language (e.g.,
Sy’s client) while retaining as many typing guarantees as possible? Sy’s client) while retaining as many typing guarantees as possible? One answer would be to use a dependently typed system that can describe the type of the
serveOp function adequately. In an extension of a recently proposed system (Toninho &
Yoshida, 2018) with iteration on natural numbers and large elimination, we might write
that code as follows: Op :
nat →Type
Op 0 = i n t
Op ( n+1) = i n t
→Op n Ch :
nat →
S e s s i o n Ch 0 = ! i n t . end! Ch ( n+1) = ? i n t . Ch n https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Camb 8 A. Igarashi et al. serveOpDep
:
( n
:
nat )
( op
: Op n )
( c
: Ch n ) →
unit
serveOpDep 0 op c = close
(send op c )
serveOpDep
( n+1) op c = l e t
v , c = receive
c
in
serveOpDep n ( op v )
c serveOpDep
:
( n
:
nat )
( op
: Op n )
( c
: Ch n ) →
unit
serveOpDep 0 op c = close
(send op c )
serveOpDep
( n+1) op c = l e t
v , c = receive
c
in
serveOpDep n ( op v )
c However, we are not aware of a fully developed theory of a session-type system that would
be able to process this definition. An alternative that is immediately available is to resort to gradual typing. For this par-
ticular program, it will insert casts to make the program type check, but all those casts
are semantically guaranteed to succeed because it would have a dependent type. To this
end, we rewrite the function dynServer in a gradually typed external language analogous to
the gradually typed lambda calculus GTLC (Siek et al., 2015b), but extended with GV’s
communication operations. 2.4 The gradual way In our example, the rewrite to the external language boils down to providing suitable
type signatures for dynServer and serveOp: dynServer
:
Compute →
unit
serveOp
:
i n t
→⋆→⋆⃝→
unit The first argument n of dynServer is consistently handled as an integer, so its type is int. The
second argument op is invoked with values of type int →int →int, int →int, and int:
these types are subsumed to the dynamic type ⋆. Similarly to other gradual type systems, an
expression of type ⋆can be used in any context, e.g., addition, function application, or even
communication, and any value can be passed where ⋆is expected. The third argument c is
invoked with channels of different types: ?int.? int .! int .end!, ?int .! int .end!, and ! int . end!. These types are subsumed to a type that is novel to this work, the dynamic session
type, ⋆⃝, a linear type which subsumes all session types. It is important to see that the
channel c is handled linearly in functions dynServer and serveOp. For that reason, the role
and handling of the linear dynamic session type with respect to the set of session types is
analogous to the role and handling of ⋆with respect to general types, as shown in earlier
work (Fennell & Thiemann, 2012; Thiemann, 2014). Aside from the type annotation, the
code remains exactly the same as in the unityped case. The external language comes with a translation into a blame calculus with explicit casts. This translation inserts just the casts that are necessary to make typing of the code go
through. Here is the output of this translation (suffix Cast is appended to the names of the
functions to distinguish different versions): dynServerCast
:
Compute →
unit
dynServerCast
c =
case c of {
neg :
c . serveOpCast 1
(( λx.−x )
:
i n t
→
i n t
ℓ1
⇒⋆)
( c
:
? i n t . ! i n t . end! ℓ2
⇒⋆⃝) ;
add :
c . serveOpCast 2
(( λx . λy . x+y )
:
i n t
→
i n t
→
i n t
ℓ3
⇒⋆)
( c
:
? i n t . ? i n t . ! i n t . end! 2.4 The gradual way ℓ4
⇒⋆⃝)
} dynServerCast
:
Compute →
unit
dynServerCast
c =
case c of {
neg :
c . serveOpCast 1
(( λx.−x )
:
i n t
→
i n t
ℓ1
⇒⋆)
( c
:
? i n t . ! i n t . end! ℓ2
⇒⋆⃝) ;
add :
c . serveOpCast 2
(( λx . λy . x+y )
:
i n t
→
i n t
→
i n t
ℓ3
⇒⋆)
( c
:
? i n t . ? i n t . ! i n t . end! ℓ4
⇒⋆⃝)
} dynServerCast
:
Compute →
unit
dynServerCast
c =
case c of {
neg :
c . serveOpCast 1
(( λx.−x )
:
i n t
→
i n t
ℓ1
⇒⋆)
( c
:
? i n t . ! i n t . end! ℓ2
⇒⋆⃝) ;
add :
c . serveOpCast 2
(( λx . λy . x+y )
:
i n t
→
i n t
→
i n t
ℓ3
⇒⋆)
( c
:
? i n t . ? i n t . ! i n t . end! ℓ4
⇒⋆⃝)
} serveOpCast
:
i n t
→⋆→⋆⃝→
unit serveOpCast
:
i n t
→⋆→⋆⃝→
unit serveOpCast
:
i n t
→⋆→⋆⃝→
unit https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at ms. https://doi.org/10.1017/S0956796819000169
idge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Camb 9 Gradual session types serveOpCast n op c =
i f
n==0 then serveOpCast n op c =
i f
n==0 then serveOpCast n op c =
i f
n==0 then
close
(( send op ( c
:
⋆⃝
ℓ5
⇒
! ⋆. ⋆⃝) )
:
⋆⃝
ℓ6
⇒
end! )
else
l e t
v , c = receive
( c
:
⋆⃝
ℓ7
⇒?⋆. ⋆⃝)
in
serveOpCast
(n−1)
(( op
:
⋆
ℓ8
⇒⋆→⋆) v )
c close
(( send op ( c
:
⋆⃝
ℓ5
⇒
! ⋆. ⋆⃝) )
:
⋆⃝
ℓ6
⇒
end! 2.5 Dynamic linearity The refined criteria for gradual typing (Siek et al., 2015b) postulate that a gradual type
system should come with a full embedding of a unityped calculus. This embedding (which
we indicate by ceiling brackets ⌈. . . ⌉) extends the embedding given for the simply typed
lambda calculus (Wadler & Findler, 2009) to handle the operations on sessions (see
Figure 13 for its definition). For example, (the unityped version of) the dynServer as written by Rob is compiled and
embedded into the gradually typed language as a value dynServer : ⋆. To directly incorpo-
rate Rob’s code, the gradual type checker enables Sy to write a function callDynServer that
accepts a channel of type Compute and returns a value of type unit, but internally just calls
dynServer. c a l l D y n S e r v e r
:
Compute →
unit
c a l l D y n S e r v e r
c =
dynServer
c c a l l D y n S e r v e r
:
Compute →
unit
c a l l D y n S e r v e r
c =
dynServer
c c a l l D y n S e r v e r
:
Compute →
unit
c a l l D y n S e r v e r
c =
dynServer
c The gradual type checker translates the definition of callDynServer by inserting the appro-
priate casts: it casts the embedded dynServer (of type ⋆) to the function type ⋆→⋆, it casts
the channel argument to this function to ⋆, and it casts the result to unit. 2.4 The gradual way )
else Casts of the form e : T1
p⇒T2—meaning that e of type T1 is cast to T2—are inserted where
values are converted from/to ⋆or ⋆⃝, similarly to the translation from GTLC. The blame
labels ℓ1, ℓ2, . . . (ranged over by p and q) on the arrow identify casts, when they fail. The
resulting casts in dynServerCast and serveOpCast look fairly involved, but we should keep in
mind that the programmer does not have to write them as they result from the translation. In
practice, blame labels may contain information on program locations to help identify how
a program fails. For example, if Rob made the following mistake in writing his dynServer neg :
c . serveOp 2 (λx.−x )
c ; −−The
f i r s t
argument
to
serveOp
should
be
1! then a call to negationClient would fail after the server receives the first integer from the
client. More specifically, the failure would identify the cast labeled ℓ7 failed because a
channel endpoint whose session type is !int.end! had been flown from ℓ2. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core t (( c
:
Compute
ℓ9⇒⋆⃝)
:
⋆⃝
ℓ9⇒⋆) The first part is a cast among linear (session) types and it can be handled as outlined in
Section 2.4. The second part is a cast from a linear type (which could be a session type, a
linear function type, or a linear product) to the unrestricted dynamic type ⋆. A cast from a linear type to unrestricted ⋆is a novelty of Gradual GV. Operationally, the
cast introduces an indirection through a store: it takes a linear value as an argument, allo-
cates a new cell in the store, moves the linear value along with a representation of its type
into the cell, and returns a handle a to the cell as an unrestricted value of type ⋆. Gradual
GV represents the cell by a process and creates handles by introducing an appropriate
binder so that a process of the form E[v : ⋆⃝
p⇒⋆] reduces to (νa)(E[a] | a →v : ⋆⃝
p⇒⋆). Here, (νa)P represents the scope of a fresh reference to a linear value and the process a →
v : ⋆⃝
p⇒⋆represents the cell storing v at a. Linear use of this cell is controlled at runtime
using ideas for runtime monitoring of affine types (Tov & Pucella, 2010; Padovani, 2017). p Any access to a cell comes in the guise of a cast a : ⋆
p⇒T from ⋆to another type applied
to a handle a. If the first access to the cell is a cast from ⋆to a linear type consistent with the
type representation stored in the cell, then the cast returns the linear value and empties the
cell. Any subsequent access to the same cell results in a linearity violation which allocates
blame to the label on the cast from ⋆. If the first cast attempts to convert to an inconsistent
type, then blame is allocated to that cast. In addition, there is a garbage collection rule that
fires when the handle of a full cell is no longer reachable from any process. It allocates
blame to the context of the cast to ⋆because that cast violated the linearity protocol by
dismissing the handle. 2.5 Dynamic linearity c a l l D y n S e r v e r
:
Compute →
unit
c a l l D y n S e r v e r
c =
(( dynServer
:
⋆
ℓ9
⇒⋆→⋆)
( c
:
Compute
ℓ10
⇒⋆) )
:
⋆
ℓ11
⇒
unit c a l l D y n S e r v e r
:
Compute →
unit
c a l l D y n S e r v e r
c =
(( dynServer
:
⋆
ℓ9
⇒⋆→⋆)
( c
:
Compute
ℓ10
⇒⋆) )
:
⋆
ℓ11
⇒
unit The casts inserted in this code make Sy’s expectations completely obvious: dynServer must
be a function and it is expected to use c as a channel of type Compute. Any misuse will
allocate blame to the respective cast in dynServer. 10 A. Igarashi et al. One kind of misuse that we have not discussed, yet, is compromising linearity: Sy has
no guarantee that Rob’s code does not accidentally duplicate or drop the communication
channel. Both actions can lead to protocol violations, which should be detected at runtime. Gradual GV takes care of linearity by factoring the cast (c : Compute
ℓ9⇒⋆) through the
dynamic session type ⋆⃝: (( c
:
Compute
ℓ9⇒⋆⃝)
:
⋆⃝
ℓ9⇒⋆) 2.6 End-to-end dynamicity The examples so far tacitly assume that channels are created with a fully specified session
type that provides a “ground truth” for the protocol on this channel. Later on, channels
may be cast to ⋆⃝and on to ⋆, but essentially they adhere to the ground truth established at
their creation. Unfortunately, this view cannot be upheld in a calculus that is able to embed a unityped
language like Uni GV. When writing new in a unityped program to create a channel, Rob
(hopefully) has some session type in mind, but it is not manifest in the code. In the typed setting, new returns a linear pair of session endpoints of type S ×lin S where
S is the server session type and S its dual client counterpart (cf. the Compute and ComputeD
types in Sections 2.1 and 2.2). When embedding the unityped new, the session type S is
unknown. Hence, the embedding needs to create a channel without an inherent ground
truth session type. It does so by assigning both channel ends type ⋆⃝and casting it to ⋆as in
new: ⋆⃝×lin ⋆⃝⇒⋆. To make this work, the dynamic session type ⋆⃝is considered self-
dual, that is ⋆⃝= ⋆⃝. Gradual GV offers no static guarantees for either end of such a
channel. 11 Gradual session types To see what runtime guarantees Gradual GV can offer for a channel of unknown session
type, let’s consider the embedding of the dynamic send and receive operations that may be
applied to it. The embedded send operation takes two arguments of type ⋆, for the value
and the channel, and returns the updated channel wrapped in type ⋆. The embedded receive
operation takes a wrapped channel of type ⋆and returns a (⋆-wrapped) pair of the received
value and the updated channel. ⌈send e f ⌉= (send ⌈e⌉(⌈f ⌉: ⋆
p⇒!⋆. ⋆⃝)): ⋆⃝⇒⋆
⌈receive e⌉= (receive (⌈e⌉: ⋆
q⇒?⋆. ⋆⃝)): ⋆×lin ⋆⃝⇒⋆ (Here, p and q are metavariables ranging over blame labels.) Now consider running the
following unityped program with entry point main. (Here, p and q are metavariables ranging over blame labels.) Now consider running the
following unityped program with entry point main. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 2.6 End-to-end dynamicity 1
c l i e n t
cc =
2
l e t
v , cc = receive
cc
in
wait
cc
3
s e r v e r
cs =
4
l e t
cs = send 42
cs
in
close
cs
5
main =
6
l e t
cs , cc = new in
7
l e t _ = fork
( c l i e n t
cc )
in
8
s e r v e r
cs 1
c l i e n t
cc =
2
l e t
v , cc = receive
cc
in
wait
cc
3
s e r v e r
cs =
4
l e t
cs = send 42
cs
in
close
cs
5
main =
6
l e t
cs , cc = new in
7
l e t _ = fork
( c l i e n t
cc )
in
8
s e r v e r
cs After a few computation steps, it reaches a configuration where the client and the server
have reduced to (νcc, cs)( client | server) where client = ⟨E[(receive (cc: ⋆⃝
q⇒?⋆. ⋆⃝)): ⋆×lin ⋆⃝⇒⋆]⟩
server = ⟨F[(send (42 : int ⇒⋆) (cs: ⋆⃝
p⇒!⋆. ⋆⃝)): ⋆⃝⇒⋆]⟩ for some contexts E and F. The channel ends cc : ⋆⃝and cs : ⋆⃝are the two ends of the
channel created in line 6. Fortunately, the two processes use the channel consistently as
the cast target ?⋆. ⋆⃝on one end is dual to the cast target !⋆. ⋆⃝at the other end. Hence,
Gradual GV has a reduction that drops the casts at both ends in this situation, and retypes
the ends to cc : ?⋆. ⋆⃝and cs : !⋆. ⋆⃝, respectively. ⟨E[(receive cc): ⋆×lin ⋆⃝⇒⋆]⟩| ⟨F[(send (42 : int ⇒⋆) cs): ⋆⃝⇒⋆]⟩ Implementing this reduction requires communication between the two processes to check
the cast targets for consistency. While our formal presentation abstracts over this imple-
mentation issue, we observe that a single asynchronous message exchange is sufficient:
Each cast first sends its target type and then receives the target type of the cast at the other
end. Then both processes check locally whether the target types are duals of one another. If they are, then both processes continue; otherwise they allocate blame. As both ends
perform the same comparison, the outcome is the same in both processes. 3 GV and gradual GV
3.1 GV We begin by discussing a language GV with session types but without gradual types. The
language is inspired by both the Gay and Vasconcelos’ (2010) functional session type
calculus and Wadler’s (2012, 2014) ‘good variant’ of the language. A main difference
from the former is the introduction of communication primitives and session types to close 12
A. Igarashi et al. Fig. 1. Types and subtyping in GV. 12
A. Igarashi et al. Fig. 1. Types and subtyping in GV. A. Igarashi et al. 12 12
A. Igarashi et al. Fig. 1. Types and subtyping in GV. Fig. 1. Types and subtyping in GV. Fig. 1. Types and subtyping in GV. Fig. 1. Types and subtyping in GV. a session explicitly. Unlike the latter, types are “stratified” into two levels—sessions types
and plain types—and deadlock freedom is not guaranteed. a session explicitly. Unlike the latter, types are “stratified” into two levels—sessions types
and plain types—and deadlock freedom is not guaranteed. 3.1.1 Types and subtyping Figure 1 summarizes types of GV. Let m, n range over multiplicities for types whose use
is either unrestricted, un, or must be linear, lin. Figure 1 summarizes types of GV. Let m, n range over multiplicities for types whose use
is either unrestricted, un, or must be linear, lin. Let T, U range over types, which include unit type, unit; unrestricted and linear function
types, T →m U; unrestricted and linear product types, T ×m U; and session types. One
might also wish to include Booleans or base types, but we omit these as they can be dealt
with analogously to unit. Let l range over labels used for selection and case choices. Let S, R range over session
types that describe communication protocols for channel endpoints, which include: send
!T. S, to send a value of type T and then behave as S; receive ?T. S, to receive a value of
type T and then behave as S, select ⊕{li : Si}i∈I, to send one of the labels li and then behave
as Si; case &{li : Si}i∈I to receive any of the labels li and then behave as Si; close end!, to
close a channel endpoint; and wait end?, to wait for the other end of the channel to close. In ⊕{li : Si}i∈I and &{li : Si}i∈I, the label set must be non-empty. We will call the session
type that describes the behavior after send, receive, select, or case the residual. 13 Gradual session types We define the usual notion of the dual of a session type S, written as S. Send is dual to
receive, select is dual to case, and close is dual to wait. Duality is an involution, so that
S = S. Multiplicities are ordered by un <: lin, indicating that an unrestricted value may be used
where a linear value is expected, but not conversely. The unit type is unrestricted, session
types are linear, while function types T →m U and product types T ×m U are unrestricted
or linear depending on the multiplicity m that decorates the type constructor. To ensure that
linear objects are used exactly once our type system imposes the invariant that unrestricted
data structures do not contain linear data structures. As an example, type unit ×un end! cannot be introduced in any derivation. 3.1.1 Types and subtyping We also write n:>(T) if m(T) holds for some m
such that m <: n, thus un:>(T) holds only if un(T), while lin:>(T) holds if either lin(T) or
un(T), and hence holds for any type. We define subtyping as usual for functional-program like systems (Gay & Vasconcelos,
2010). Function types are contravariant in their domain, covariant in their range, and
covariant in their multiplicity, and send types are contravariant in the value sent and
covariant in the residual session type. All other types and session types are covariant in
all components. Width subtyping resembles record subtyping for select, and variant sub-
typing for case. That is, on an endpoint where one may select among labels with an index
in I one may instead select among labels with indexes in J, so long as J ⊆I, while on
an endpoint where one must be able to receive any label with an index in I one may
instead receive any label with an index in J, so long as I ⊆J. (Subtyping on endpoints
appears sometimes reversed in process-calculus like systems, such as (Carbone et al., 2007,
2012; Demangeon & Honda, 2011), Wadler’s CP (2012, 2014); Gay (2016) discusses the
situation.) Subtyping is reflexive, transitive, and antisymmetric. Duality inverts subtyping, in that
S <: R if and only if R <: S. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 3.1.2 Expressions, processes, and typing 3.1.2 Expressions, processes, and typing 3.1.2 Expressions, processes, and typing Expressions, processes, and typing for GV are summarized in Figure 2. We let x, y range
over variables, c, d range over channel endpoints, and z range over names, which are either
variables or channel endpoints. We let e, f range over expressions, which include names, unit value, function abstraction
and application, pair creation and destruction, fork a process, create a new pair of chan-
nel endpoints, send, receive, select, case, close, and wait. Function abstraction and pair
creation are labeled with the multiplicity of the value created. We sometimes abbreviate
expressions of the form (λlinx.e)f to let x = e in f , as usual. A GV program is always given
as an expression, but as it executes it may fork new processes. We let P, Q range over processes, which include expressions, parallel composition, and
a binder that introduces a pair of channel endpoints. The initial process will consist of a
single expression, corresponding to a given GV program. The bindings in the language are as follows: variable x is bound in subexpression e of
λmx.e, variables x, y are bound in subexpression f of let x, y = e in f , variables xi are bound
in subexpressions ei of case e of {li : xi.ei}i∈I, channel endpoints c, d are bound in subpro-
cess P of (νc, d)P. We assume that c and d in (νc, d)P are different. The notions of free and
bound names/variables as well that substitution are defined accordingly. The set of the free 14
A. Igarashi et al. Fig. 2. Expressions, processes, and typing in GV. A. Igarashi et al. 14 Fig. 2. Expressions, processes, and typing in GV. Fig. 2. Expressions, processes, and typing in GV. names in P is denoted by fn(P). We follow Barendregt’s variable convention, whereby
all names in binding occurrences in any mathematical context are pairwise distinct and
distinct from the free names (Barendregt, 1984). We let , range over environments, which are used for typing. An environment con-
sists of zero or more associations of names with types. Environment splitting = 1 ◦2
is standard. 3.1.2 Expressions, processes, and typing It breaks an environment for an expression or process into environments 1
and 2 for its components; a name of unrestricted type may be used in both environments,
while a name of linear type must be used in one environment or the other but not both. We
write m() if m(T) holds for each T in , and similarly for m:>(). Write ⊢e : T if under environment expression e has type T. The typing rules for
expressions are standard. In the rules for names, unit, and new the remaining environment
must be unrestricted, to enforce the invariant that linear variables are used exactly once. A function abstraction that is unrestricted must have only unrestricted variables bound
in its closure, and a pair that is unrestricted may only contain components that are unre-
stricted. Thus, it is never possible to construct a pair of type, e.g., S ×un T, which contains ms. https://doi.org/10.1017/S0956796819000169
idge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 15 Gradual session types Gradual session types
15
Fig. 3. Reduction in GV. Fig. 3. Reduction in GV. Fig. 3. Reduction in GV. a linear type S under the unrestricted pair type constructor ×un, even though such a type
is syntactically allowed for simplicity. The rules for send, receive, select, case, close, and
wait match the corresponding session types. For example, the following type judgment o: int, c: !int.end! ⊢close (send o c) : unit o: int, c: !int.end! ⊢close (send o c) : unit can be derived. The typing system supports subsumption: if e has type T and T is a subtype
of U then e also has type U. can be derived. The typing system supports subsumption: if e has type T and T is a subtype
of U then e also has type U. Write ⊢P if under environment process P is well typed. The typing rules for pro-
cesses are also standard. If expression e has unrestricted type T then process ⟨e⟩is well
typed. If processes P and Q are well typed, then so is process P | Q, where the environment
of the latter can be split to yield the environments for the former. 3.1.2 Expressions, processes, and typing And if process P is well-
typed under an environment that includes channel endpoints c and d with session types S
and S, then process (νc, d)P is well typed under the same environment without c and d. A. Igarashi et al. Write e −→f to indicate that expression e reduces to expression f . Reduction is
standard, consisting of beta reduction for functions and pairs. Write e −→f to indicate that expression e reduces to expression f . Reduction is
standard, consisting of beta reduction for functions and pairs. Write P ≡Q for structural congruence of processes. It is standard, with composition
being commutative and associative. A process returning the unit is the identity of parallel
composition, so P | ⟨()⟩≡P. The order in which the endpoints are written in a ν-binder
is irrelevant. Distinct prefixes commute, and satisfy scope extrusion. The Barendregt con-
vention ensures that c, d are not free in Q in the rule for scope extrusion. Similarly for the
rule to swap prefixes. Write P −→Q if process P reduces to process Q. Evaluating fork e returns () and
creates a new process ⟨e⟩. Evaluating new introduces a new binder (νc, d) and returns
a pair (c, d)lin of channel endpoints. Evaluating send v c on one endpoint of a channel
and receive d on the other, causes the send to return c and the receive to return (v, d)lin. Similarly for select on one endpoint of a channel and case on the other, or close on one
endpoint of a channel and wait on the other. Process reduction is a congruence with regard to parallel composition and binding
for channel endpoints, it is closed under structural congruence, and supports expression
reduction under evaluation contexts. 3.2 Gradual GV We now introduce Gradual GV. Following standard frameworks of gradual typing (Siek &
Taha, 2006; Siek et al., 2015b), Gradual GV consists of two sublanguages: an external
language GGVe, in which source programs are written, and an internal language GGVi, to
which GGVe is elaborated by cast-inserting translation to make necessary runtime checks
explicit. The operational semantics of a program is given as reduction of processes in
GGVi. We first introduce GGVi by outlining its differences to GV (Sections 3.2.1–3.2.3). Next, we introduce the syntax of GGVe, which has only expressions, because it is the lan-
guage in which source programs are written, its type system, and cast-inserting translation
from GGVe to GGVi (Sections 3.2.4 and 3.2.5). Finally, we discuss how an untyped variant
of GV can be embedded into GGVi (Section 3.2.6). 3.1.3 Reduction Values, evaluation contexts, reduction for expressions, structural congruence, and reduc-
tion for processes for GV are summarized in Figure 3. Let v, w range over values, which include unit, function abstractions, pairs of values,
and channel endpoints. Let E, F range over evaluation contexts, which are standard. ms. https://doi.org/10.1017/S0956796819000169
idge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 16 https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 3.2.1 Types and subtyping Following the usual approach to gradual types, we extend the grammar of types with a
dynamic type (sometimes also called the unknown type), written ⋆. Similarly, we extend
session types with the dynamic session type, written ⋆⃝. The extended grammar of types
is given in Figure 4, where types carried over from Figure 1 are typeset in gray. As before, we let T, U range over types and S, R range over session types. We also
distinguish a subset of types which we call ground types, ranged over by T, U, and a subset
of session types which we call ground session types, ranged over by S, R, consisting of all
the type constructors applied only to arguments which are either the dynamic type or the
dynamic session type, as appropriate. We define ⋆⃝to be self-dual: ⋆⃝= ⋆⃝. We define the multiplicity of the new types by
setting ⋆to be un and ⋆⃝to be lin. The remaining definitions of multiplicity of types carries
over unchanged from Figure 1. Type ⋆is labeled unrestricted although (as we will see in the
following) it corresponds to all possible types, both unrestricted and linear, and therefore Gradual session types
17
Fig. 4. Types and subtyping in Gradual GV. Gradual session types 17 Fig. 4. Types and subtyping in Gradual GV. Fig. 4. Types and subtyping in Gradual GV. we will need to take special care when handling values of type ⋆that correspond to values
of a linear type. Consistent subtyping is defined over types of Gradual GV also in Figure 4. It is identical
to the definition of subtyping from Figure 1, with each occurrence of <: replaced by ≲,
and with the addition of four rules for the new types: ⋆≲T
T ≲⋆
⋆⃝≲S
S ≲⋆⃝ For example, we have (a) ⊕{l1 : !⋆. ⋆⃝, l2 : ?⋆. ⋆⃝} ≲⊕{l1 : ⋆⃝} and (b) &{l1 : ⋆⃝} ≲&{l1 :
!⋆. ⋆⃝, l2 : ?⋆. ⋆⃝}. Consistent subtyping is reflexive, but neither symmetric nor transitive. As with subtyping, we have S ≲R iff R ≲S. In Gradual GV, we will be permitted to
attempt to cast a value of type T to a value of type U exactly when T ≲U. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 3.2.1 Types and subtyping A cast may
fail at runtime: while a cast using (a) will not fail, a cast using (b) may fail because an
expression of type &{l1 : ⋆⃝} may evaluate to a value of type, say, &{l1 : end!}. For example, we have (a) ⊕{l1 : !⋆. ⋆⃝, l2 : ?⋆. ⋆⃝} ≲⊕{l1 : ⋆⃝} and (b) &{l1 : ⋆⃝} ≲&{l1 :
!⋆. ⋆⃝, l2 : ?⋆. ⋆⃝}. Consistent subtyping is reflexive, but neither symmetric nor transitive. As with subtyping, we have S ≲R iff R ≲S. In Gradual GV, we will be permitted to
attempt to cast a value of type T to a value of type U exactly when T ≲U. A cast may
fail at runtime: while a cast using (a) will not fail, a cast using (b) may fail because an
expression of type &{l1 : ⋆⃝} may evaluate to a value of type, say, &{l1 : end!}. Two types are consistent, written T ∼U, if T ≲U and U ≲T. Consistency is reflexive
and symmetric but not transitive. The standard example of the failure of transitivity is that
for any function type we have T →m U ∼⋆and for any product type we have ⋆∼T′ ×n U′,
but T →m U ̸∼T′ ×n U′. In the setting of session types one has for example ?T. S ∼⋆⃝and
⋆⃝∼end!, but ?T. S ̸∼end!. Subtyping T <: U for Gradual GV essentially carries over from GV. Its definition is
exactly as in Figure 1, with the addition of two rules that ensure subtyping is reflexive
for the dynamic type and the dynamic session type. In contrast to consistent subtyping,
subtyping T <: U guarantees that we may always treat a value of the first type as if it
belongs to the second type without casting. A. Igarashi et al. 18 18
A. Igarashi et al. Fig. 5. Expressions, processes, and typing in GGVi. Fig. 5. Expressions, processes, and typing in GGVi. Fig. 5. Expressions, processes, and typing in GGVi. Fig. 5. Expressions, processes, and typing in GGVi. Fig. 5. Expressions, processes, and typing in GGVi. blame p q X
or
blame p X where p and q are blame labels, and X is a set of variables of linear type. As we will see,
most instances that yield blame involve two casts, hence the form with two blame labels,
although blame can arise for a single cast, hence the form with one blame label. The set
X records all linear variables in scope when blame is raised, and is used to maintain the
invariant that as a program executes each variable of linear type appears linearly (only
once, or once in each branch of a case). Discarding linear variables when raising blame
would break the invariant. Blame corresponds to raising an exception, and the list of linear
variables corresponds to cleaning up after linear resources when raising an exception (for
instance, closing an open file or channel). In the typing rules, the notation flv() refers the
set of free variables of linear type that appear in . We also write flv(E) and flv(v) for the
free linear variables appearing in an evaluation context E or a value v. In a running pro-
gram, only free linear variables are channel endpoints, so flv(E) and flv(v) can be defined
without type information. where p and q are blame labels, and X is a set of variables of linear type. As we will see,
most instances that yield blame involve two casts, hence the form with two blame labels,
although blame can arise for a single cast, hence the form with one blame label. The set
X records all linear variables in scope when blame is raised, and is used to maintain the
invariant that as a program executes each variable of linear type appears linearly (only
once, or once in each branch of a case). Discarding linear variables when raising blame
would break the invariant. Blame corresponds to raising an exception, and the list of linear
variables corresponds to cleaning up after linear resources when raising an exception (for
instance, closing an open file or channel). In the typing rules, the notation flv() refers the
set of free variables of linear type that appear in . We also write flv(E) and flv(v) for the
free linear variables appearing in an evaluation context E or a value v. In a running pro-
gram, only free linear variables are channel endpoints, so flv(E) and flv(v) can be defined
without type information. blame p q X
or
blame p X The processes of GGVi are those of GV, plus three additional forms for references to
linear values (as well as blame, described above). Recall that a value of type ⋆may contain
a linear value, in which case dynamic checking must ensure that it is used exactly once. The
mechanism for doing so is to allocate a reference to a linear value. We let a, b range over
references. A reference is of type ⋆, and contains a value w of ground type T, where T is
linear (either ⋆→lin ⋆or ⋆×lin ⋆or the dynamic session type ⋆⃝). References are allocated
by the binding form (νa)P, and the value contained in store a is indicated by a process
which is either of the form a →w : T
p⇒⋆
or
a →locked p where w is a value of type T and p is a blame label. Bindings for references initially take
the first form, but change to the second form after the reference has been accessed once;
any subsequent attempt to access the reference a second time will cause an error. where w is a value of type T and p is a blame label. Bindings for references initially take
the first form, but change to the second form after the reference has been accessed once;
any subsequent attempt to access the reference a second time will cause an error. where S, R ̸= ⋆⃝. 3.2.2 Expressions, processes, and typing of GGVi Blame is indicated by processes of the form Blame is indicated by processes of the form blame p q X
or
blame p X 3.2.2 Expressions, processes, and typing of GGVi Expressions, processes, and type rules of GGVi are summarized in Figure 5. The
expressions of GGVi are those of GV, plus an additional form for casts. A cast is written e : T
p⇒U
(1) e : T
p⇒U (1) where e is an expression of type T, and p, q range over blame labels such as ℓ1, ℓ2, . . .. For
example, the following term SOC = λuno.λunc.close ((send o (c: ⋆⃝
ℓ1⇒!⋆. ⋆⃝)): ⋆⃝
ℓ2⇒end!) which represents a simplified version of serveOpCast in Section 2, can be given type ⋆→un
⋆⃝→un unit. Blame labels carry a polarity, which is either positive or negative. The complement
operation, p, takes a positive label into a negative one and vice versa; complement is an
involution, so that p = p. By convention, we assume that all blame labels in a source pro-
gram are positive, but negative blame labels may arise during evaluation of casts at a
function type or a send type. A cast raises positive blame if the fault lies with the expres-
sion contained in the cast (for instance, because it returns an integer where a character is
expected), while it raises negative blame if the fault lies with the context containing the
cast (for instance, because it passes an argument or sends a value that is an integer where
a character is expected). In a valid cast e : T
p⇒U, the type T must be a consistent subtype of U (T ≲U), the
type of the entire expression. If a cast in a program fails, it evaluates to blame p q X
or blame p X (which, as we see later, are treated as processes) where the blame label
p and q indicate the root cause of the failure (we will explain X shortly). If the cast
in (1) fails, it means that the value returned by e has type T, but not type U. For exam-
ple, let e = 4711 : int
q⇒⋆, T = ⋆, and U = bool. As ⋆≲bool, the resulting expression 19 Gradual session types (4711 : int
q⇒⋆) : ⋆
p⇒bool is well typed. However, at runtime it raises blame by reducing
to blame q p ∅, which flags the error that int is not a subtype of bool: that is, int ̸<: bool. 3.2.3 Reduction Values, evaluation contexts, reductions for expressions, structural congruence, and reduc-
tions for processes for GGVi are summarized in Figures 6 and 7. The values of GGVi are those of GV, plus five additional forms. Values of dynamic type
have the form either v : T
p⇒⋆as in other blame calculi, if T is unrestricted, or a, which is
a reference to a linear value, if the dynamic type wraps a linear value. Additionally, there
are values of dynamic session type which take the form v : S
p⇒⋆⃝. Following standard practice for blame calculus, we take a cast of a value between func-
tion types to be a value, and for similar reasons a cast from a session type to a session type
is a value unless one end of the cast is the dynamic session type: v : T →m U
p⇒T′ →n U′
or
v : S
p⇒R where S, R ̸= ⋆⃝. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at A. Igarashi et al. 20 Fi
6
R d
ti
i GGV
i Fig. 6. Reduction in GGVi, expressions. Fig. 7. Reduction in GGVi, processes. Fig. 7. Reduction in GGVi, processes. Fig. 7. Reduction in GGVi, processes. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at ms. https://doi.org/10.1017/S0956796819000169
idge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Camb 21 Gradual session types Additional reductions for expressions appear in Figure 6. Typical of blame calculus is
the reduction for a cast between function types, often called the wrap rule: Additional reductions for expressions appear in Figure 6. Typical of blame calculus is
the reduction for a cast between function types, often called the wrap rule: (v : T →m U
p⇒T′ →n U′) w −→(v (w: T′
p⇒T)): U
p⇒U′ The cast on the function decomposes into two casts, one on the domain and one on the
range. 3.2.3 Reduction The fact that subtyping (and consistent subtyping) for function types is contravariant
on the domain and covariant on the range is reflected in the fact that the cast on the domain
is from T′ to T and complements the blame label p, while the cast on the range is form U
to U′ and leaves the blame label p unchanged. Casts for products follow a similar pattern,
though covariant on all components. The cast on the function decomposes into two casts, one on the domain and one on the
range. The fact that subtyping (and consistent subtyping) for function types is contravariant
on the domain and covariant on the range is reflected in the fact that the cast on the domain
is from T′ to T and complements the blame label p, while the cast on the range is form U
to U′ and leaves the blame label p unchanged. Casts for products follow a similar pattern,
though covariant on all components. Reductions on session types follow the pattern of the reduction for a cast between send
types: send v (w: !T. S
p⇒!T′. S′) −→(send (v : T′
p⇒T) w): S
p⇒S′ The cast on the send decomposes into two casts, one on the value sent and one on the
residual session type. The fact that subtyping (and consistent subtyping) for send types is
contravariant on the value sent and covariant on the residual session type is reflected in
the fact that the cast on the value sent is from T′ to T and complements the blame label
p, while the cast on the residual session type is from S to S′ and leaves the blame label
p unchanged. The casts for the remaining session types follow a similar pattern, though
covariant on all components. Also typical of blame calculus, casts to the dynamic type factor through a ground type, Also typical of blame calculus, casts to the dynamic type factor through a ground type,
v : T
p⇒⋆−→(v : T
p⇒T): T
p⇒⋆ v : T
p⇒⋆−→(v : T
p⇒T): T
p⇒⋆ when T ̸= ⋆, T ̸= T, and T ∼T. This factoring is unique because for every type T such that
T ̸= ⋆there is a unique ground type T such that T ∼T. The additional condition T ̸= T
ensures that the factoring is nontrivial and that reduction does not enter a loop. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 3.2.3 Reduction Casts from
the dynamic type, and casts to and from the dynamic session type are handled analogously. when T ̸= ⋆, T ̸= T, and T ∼T. This factoring is unique because for every type T such that
T ̸= ⋆there is a unique ground type T such that T ∼T. The additional condition T ̸= T
ensures that the factoring is nontrivial and that reduction does not enter a loop. Casts from
the dynamic type, and casts to and from the dynamic session type are handled analogously. Additional structural congruences and reductions for processes appear in Figure 7. Like
bindings for channel endpoints, bindings for references to linear values satisfy scope
extrusion and reduction is a congruence with respect to them. The first five reduction rules for processes deal with references to linear values, ensuring
that a value cast from a linear type to ⋆is accessed exactly once. As the only values of the
dynamic type are casts from a ground type, expressions of interest take the form v : T
p⇒⋆ where v is a value and T is a linear ground type. The first rule introduces a reference,
represented as a separate process of the form a →v : T
p⇒⋆. The context restriction
E ̸= F[[ ]: ⋆
q⇒U] ensures that a reference is only introduced if the value is not imme-
diately accessed; without the restriction this rule would apply to a process of the form
⟨E[(v : T
p⇒⋆): ⋆
q⇒U]⟩, to which the sixth or seventh rule should be applied. Any attempt
to access the linear reference a must take the form E[a: ⋆
q⇒U] where E is an evaluation context and U is a ground type that may or may not be linear. The
second rule implements the first access to a linear value by copying the value v in place 22 A. Igarashi et al. of the reference a, and updating the reference process to a →locked p, indicating that the
linear reference has been accessed once. The third rule implements any subsequent attempt
to access a linear value, which allocates blame to the two casts involved, negative blame
p from locked p, which was a cast v : T
p⇒⋆before the first access, and positive blame
q for the cast to access a, indicating that in both cases blame is allocated to the side of
the cast of type ⋆. 3.2.3 Reduction The blame term also contains flv(E), the set of free linear variables that
appear in the context E, which as mentioned earlier is required to maintain the invariant on
linear variables; all occurrences of blame contain corresponding sets of linear variables,
which we will not mention further. The final two rules indicate what happens when all
processes containing the reference finish execution. If the linear reference is locked then
it was accessed once, and the reference may be deallocated as usual. If the reference is
not locked then it was never accessed, and blame should be allocated to the context of the
original cast, which discarded the value rather than using it linearly. In practice, these rules
would be implemented as part of garbage collection. The remaining six rules come in three pairs. Typical of blame calculus is the first pair,
often called the collapse and collide rules: ⟨E[(v : T
p⇒⋆): ⋆
q⇒U]⟩−→⟨E[v]⟩
if T <: U
⟨E[(v : T
p⇒⋆): ⋆
q⇒U]⟩−→blame p q (flv(E) ∪flv(v))
if T ̸<: U If the source type is a subtype of the target type, the casts collapse to the original value. Types are preserved by subsumption: since v has type T and T <: U then v also has type U. Conversely, if the source type is not a subtype of the target type, then the casts are in colli-
sion and reduce to blame. Blame is allocated to both of the casts involved, negative blame
p for the inner cast and positive blame q for the outer cast, indicating that in both cases
blame is allocated to the side of the cast of type ⋆. Our choice to allocate blame to both
casts differs from the usual formulation of blame calculus, which only allocates blame to
the outer cast. Allocating blame to only the outer cast is convenient if one wishes to imple-
ment blame calculus by erasure to a dynamically typed language, where injection of a value
to the dynamic type is represented by the value itself, that is, the erasure of v : T
p⇒⋆is just
taken to be the erasure of v itself. However, this asymmetric implementation is less appro-
priate in our situation. For session types, a symmetric formulation is more appropriate, as
we will see shortly when we look at the interaction between casts and communication. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 3.2.3 Reduction The next pair of rules transpose collapse and collide from types to session types. The
final pair of rules adapt collapse and collide to the case of communication between two
channel endpoints. Here is the adapted collapse rule. (νc, d)(⟨E[c: ⋆⃝
p⇒S]⟩| ⟨F[d : ⋆⃝
q⇒R]⟩) −→(νc, d)(⟨E[c]⟩| ⟨F[d]⟩)
if S <: R The condition on this rule is symmetric, since S <: R if and only if R <: S. On the left-hand
side of this rule c, d both have session type ⋆⃝, while on the right-hand side of the rule c, d
have session types S, S or R, R. Again, types are preserved by subsumption, since if c, d
have session types S, S and S <: R then c, d also have session types S, R, and similarly if
c, d have session types R, R. Analogously, the last rule adapts collide. An alternative design might replace the final pair of rules by a structural congruence that
slides a cast from one endpoint of a channel to the other: (νc, d)(E[c: S
p⇒R] | F[d]) ≡(νc, d)(E[c] | F[d : R
p⇒S]). 23 Gradual session types Setting S to ⋆⃝and R to S, this congruence can reduce the third collapse rule (on chan-
nel endpoints) to the second collapse rule (on a nested pair of casts on session types). However, even with this congruence the two collide rules are not quite equivalent. Our
chosen formulation, though slightly longer, is more symmetric and easier to implement. Now we show a few examples of reduction, in which we abbreviate a nested cast
(e: T1
p⇒T2): T2
q⇒T3 to e: T1
p⇒T2
q⇒T3 and use a sequential composition e1; e2 with
obvious typing and reduction rules. First recall the term SOC = λuno.λunc.close (send o (c: ⋆⃝
ℓ1⇒!⋆. ⋆⃝): ⋆⃝
ℓ2⇒end!)
introduced above. Given a channel endpoint d : !int.end!, the term
SOC (42: int
ℓ3⇒⋆) (d : !int.end! ℓ4⇒⋆⃝) SOC = λuno.λunc.close (send o (c: ⋆⃝
ℓ1⇒!⋆. ⋆⃝): ⋆⃝
ℓ2⇒end!) introduced above. Given a channel endpoint d : !int.end!, the term
SOC (42: int
ℓ3⇒⋆) (d : !int.end! ℓ4⇒⋆⃝) introduced above. Given a channel endpoint d : !int.end!, the term
SOC (42: int
ℓ3⇒⋆) (d : !int.end! ℓ4⇒⋆⃝) introduced above. Given a channel endpoint d : !int.end!, the term
SOC (42: int
ℓ3⇒⋆) (d : !int.end! ℓ4⇒⋆⃝) SOC (42: int
ℓ3⇒⋆) (d : !int.end! (νd, e)(blame ℓ4 ℓ2 {d} | ⟨let x, y = (42, e)lin in . . .⟩) We also show an example of dynamic linearity checking. The following function foo
takes an argument of type ⋆, cast it to end!, and closes it: foo = λunx.close (x: ⋆
ℓ⇒end!) Consider an application of foo to a channel endpoint c of type end!. It reduces as follows: ⟨foo (c: end! ℓ′
⇒⋆)⟩
−→⟨foo (c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆)⟩ ⟨foo (c: end! ℓ′
⇒⋆)⟩
−→⟨foo (c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆)⟩
−→(νa)(⟨foo a⟩| a →c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆)
−→(νa)(⟨close (a: ⋆
ℓ⇒end!)⟩| a →c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆)
−→(νa)(⟨close (c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆
ℓ⇒end!)⟩| a →locked ℓ′)
−→+(νa)(⟨close c⟩| a →locked ℓ′)
−→⟨close c⟩ −→(νa)(⟨close (a: ⋆
ℓ⇒end!)⟩| a →c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆)
−→(νa)(⟨close (c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆
ℓ⇒end!)⟩| a →locked ℓ′) If the channel endpoint is passed to a function that uses the argument more than once,
blame will be raised. Let bar be λunx.close (x: ⋆
ℓ⇒end!); close (x: ⋆
ℓ⇒end!) and
observe that bar (c: end! ℓ′
⇒⋆) reduces as follows: If the channel endpoint is passed to a function that uses the argument more than once,
blame will be raised. Let bar be λunx.close (x: ⋆
ℓ⇒end!); close (x: ⋆
ℓ⇒end!) and
observe that bar (c: end! ℓ′
⇒⋆) reduces as follows: ⟨bar (c: end! ℓ′
⇒⋆)⟩
−→⟨bar (c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆)⟩ ⟨bar (c: end! ℓ′
⇒⋆)⟩
−→⟨bar (c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆)⟩
−→(νa)(⟨bar a⟩| a →c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆) −→⟨bar (c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆)⟩ −→(νa)(⟨bar a⟩| a →c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆) −→(νa)(⟨close (a: ⋆
ℓ⇒end!); close (a: ⋆
ℓ⇒end!)⟩| a →c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆)
−→(νa)(⟨close (c: end! ℓ′
⇒⋆⃝
ℓ′
⇒⋆
ℓ⇒end!); close (a: ⋆
ℓ⇒end!)⟩| a →locked ℓ′)
−→+(νa)(⟨close c; close (a: ⋆
ℓ⇒end!)⟩| a →locked ℓ′) Then, parallel composition with a process waiting at the other end d of the endpoint c will
raise blame as follows: (νc, d)(⟨bar (c: end! ℓ′
⇒⋆)⟩| ⟨wait d⟩)
−→+(νc, d)(νa)(⟨close c; close (a: ⋆
ℓ⇒end!)⟩| a →locked ℓ′ | ⟨wait d⟩))
−→(νc, d)(νa)(⟨close (a: ⋆
ℓ⇒end!)⟩| a →locked ℓ′ | ⟨()⟩)
−→(νc, d)(νa)(blame ℓ
′ ℓ∅| a →locked ℓ′) 3.2.3 Reduction ℓ4⇒⋆⃝) reduces as follows: SOC (42: int
ℓ3⇒⋆) (d : !int.end! ℓ4⇒⋆⃝)
−→(λunc.close ((send (42: int
ℓ3⇒⋆) (c: ⋆⃝
ℓ1⇒!⋆. ⋆⃝)): ⋆⃝
ℓ2⇒end!)) (d : !int.end! ℓ4⇒⋆⃝)
−→(λunc.close ((send (42: int
ℓ3⇒⋆) (c: ⋆⃝
ℓ1⇒!⋆. ⋆⃝)): ⋆⃝
ℓ2⇒end!)) SOC (42: int
ℓ3⇒⋆) (d : !int.end! ℓ4⇒⋆⃝) (d : !int.end! ℓ4⇒!⋆. ⋆⃝
ℓ4⇒⃝ −→close ((send (42: int
ℓ3⇒⋆) (d : !int.end! ℓ4⇒!⋆. ⋆⃝
ℓ4⇒⋆⃝
ℓ1⇒!⋆. ⋆⃝)): ⋆⃝
ℓ2⇒end!)
−→close ((send (42: int
ℓ3⇒⋆) (d : !int.end! ℓ4⇒!⋆. ⋆⃝)): ⋆⃝
ℓ2⇒end!)
−→close ((send (42: int
ℓ3⇒⋆
ℓ4
⇒int) d): end! ℓ4⇒⋆⃝
ℓ2⇒end!)
−→close ((send 42 d): end! ℓ4⇒⋆⃝
ℓ2⇒end!). Thus, the process Thus, the process Thus, the process Thus, the process (νd, e)(⟨SOC (42: int
ℓ3⇒⋆) (d : !int.end! ℓ4⇒⋆⃝)⟩| ⟨let x, y = receive e in wait y⟩)
reduces as follows: (νd, e)(⟨SOC (42: int
ℓ3⇒⋆) (d : !int.end! ℓ4⇒⋆⃝)⟩| ⟨let x, y = receive e in wait y⟩)
−→+(νd, e)(⟨close ((send 42 d): end! ℓ4⇒⋆⃝
ℓ2⇒end!)⟩| ⟨let x, y = receive e in wait y⟩)
−→(νd, e)(⟨close (d : end! ℓ4⇒⋆⃝
ℓ2⇒end!)⟩| ⟨let x, y = (42, e)lin in wait y⟩)
−→+(νd, e)(⟨close d⟩| ⟨wait e⟩) −→(νd, e)(⟨()⟩| ⟨()⟩) −→(νd, e)(⟨()⟩| ⟨()⟩) However, if d is given type !int.!int.end!, then SOC (42: int
ℓ3⇒⋆) (d : !int.!int.end! ℓ4⇒⋆⃝)
is well typed but reduces to close ((send 42 d): !int.end! ℓ4⇒⋆⃝
ℓ2⇒end!) close ((send 42 d): !int.end! ℓ4⇒⋆⃝
ℓ2⇒end!) Thus, the process (νd, e)(⟨SOC (42: int
ℓ3⇒⋆) (d : !int.!int.end! ℓ4⇒⋆⃝)⟩| ⟨let x, y = receive e in . . .⟩)
reduces to (νd, e)(⟨close (d : !int.end! ℓ4⇒⋆⃝
ℓ2⇒end!)⟩| ⟨let x, y = (42, e)lin in . . .⟩) https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at ms. https://doi.org/10.1017/S0956796819000169
idge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at A. Igarashi et al. 24 and then to (νd, e)(blame ℓ4 ℓ2 {d} | ⟨let x, y = (42, e)lin in . . .⟩) 3.2.4 External language GGVe Having defined the internal language, we introduce the external language GGVe, in
which source programs are written. The syntax of expressions of GGVe is presented in
Figure 8. For ease of type checking, variable declarations in functions and channel endpoint 25 Gradual session types Fig. 8. Expressions in GGVe. Fig. 8. Expressions in GGVe. Fig. 9. Matching. Fig. 9. Matching. creations are explicitly typed. There are no processes in GGVe: a program is a well-typed
closed expression and it is translated to a GGVi expression before it runs. creations are explicitly typed. There are no processes in GGVe: a program is a well-typed
closed expression and it is translated to a GGVi expression before it runs. The type system of GGVe adheres to standard practice for gradually typed languages
(Siek et al., 2015b; Cimini & Siek, 2016), but requires a few adaptations to cater for fea-
tures not covered in previous work. We first introduce a few auxiliary definitions used in
typing rules. Figure 9 defines the matching relation T ▷U (Cimini & Siek, 2016). Roughly
speaking, T ▷U means that T can be used, after necessary runtime checking, as U. The
second and third columns declare that, if T is ⋆or ⋆⃝, then it can be used as any type or
session type, respectively. Otherwise, the matching relation extracts substructure, i.e., the
domain type, the codomain type, the first-element type, and so on, from T. So, we have
neither ⋆▷unit nor ⋆⃝▷end! or ⋆⃝▷end?. Matching for the internal and external choice types is slightly involved as it has to cater
for subtyping. Matching for internal choice is invoked in the type rule for an expression
select l e. Thanks to subtyping, the type of e can be any internal choice with a branch
for label l. Hence, matching only asks for the presence of this single label and extracts its
residual. Dually, matching for external choice is invoked in the rule for a case e of . . . expression. Again due to subtyping, the case expression can check more labels than provided by the
type of e. Hence, matching allows extra branches to be checked with arbitrary residual
types (lj : Sj in the definition) while extracting the residual types for all branches provided
by e. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 3.2.4 External language GGVe Obtaining the result type of a case expression from the types of its branches requires
a join operation T ∨U that ensures that its result is (in a certain sense) a supertype of
both T and U. Figure 10 contains the definitions of join and its companion meet, which is
needed in contravariant positions of the type. Both operations are partial: join or meet is
undefined for cases other than those listed in Figure 10. Join of two ⊕-types can be obtained by taking the joins of the types associated with
common labels. Note that labels where the joins Si ∨Ri do not exist will be dropped. On
the other hand, the label set of the join of two &-types is the union of the two label sets A. Igarashi et al. 26 Fig. 10. Join and meet of types. Fig. 10. Join and meet of types. Fig. 10. Join and meet of types. Fig. 10. Join and meet of types. from the input. For the common labels in I ∩J, the joins Sk ∨Rk must exist. Join or meet
is undefined if the resulting type is ⊕{} or &{} (with the empty set of labels) as they are
ill-formed types. from the input. For the common labels in I ∩J, the joins Sk ∨Rk must exist. Join or meet
is undefined if the resulting type is ⊕{} or &{} (with the empty set of labels) as they are
ill-formed types. Without the last four clauses, which deal with ⋆and ⋆⃝, the definitions of the join and
meet coincide with those for ordinary subtyping. This is motivated by the static embedding
property of the Criteria for Gradual Typing (Siek et al., 2015b), which requires the typabil-
ity of a GGVe term without ⋆(or ⋆⃝in our case) to be the same as the typability under the
GV typing rules. There are a few choices for the join (and meet) of ⋆and other types and ms. https://doi.org/10.1017/S0956796819000169
idge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at Gradual session types 27 Fig. 11. Expression typing in GGVe. Fig. 11. Expression typing in GGVe. 3.2.4 External language GGVe we choose ⋆∨T to be T for any T because, as we prove later, our join then corresponds to
the least upper bound with respect to negative subtyping (Wadler & Findler, 2009), which
is formally defined later, and we can construct a type-checking algorithm that produces a
minimal type with respect to the negative subtyping. (The least upper bound with respect
to positive subtyping is not a good choice because int ∨bool = ⋆holds, invalidating the
static embedding property.) we choose ⋆∨T to be T for any T because, as we prove later, our join then corresponds to
the least upper bound with respect to negative subtyping (Wadler & Findler, 2009), which
is formally defined later, and we can construct a type-checking algorithm that produces a
minimal type with respect to the negative subtyping. (The least upper bound with respect
to positive subtyping is not a good choice because int ∨bool = ⋆holds, invalidating the
static embedding property.) Typing rules are presented in Figure 11. The matching relation is used in elimina-
tion rules. To obtain a syntax-directed inference system, the subsumption is merged into
function application, sending, select, and case. Moreover, subtyping is replaced with con-
sistent subtyping. The type of the whole case expression is obtained by joining the types
of the branches. Finally, the judgment e prog means that e is a Gradual GV program, which
is a closed, well-typed GGVe expression of unrestricted type. Cast insertion discussed in
the following translates a program to a GGVi expression e, which runs as a process ⟨e⟩. For
example, we can derive ⊢λuno : ⋆.λunc : ⋆⃝.close (send o c) : ⋆→un ⋆⃝→un unit. ⊢λuno : ⋆.λunc : ⋆⃝.close (send o c) : ⋆→un ⋆⃝→un unit. We also develop a type-checking algorithm for GGVe by following the stan-
dard approach (Kobayashi et al., 1999; Walker, 2005). We define an algorithm
CHECKEXPR(, e), which takes a type environment and an expression e and returns
a type T of e and the set X of linear variables in e. We avoid nondeterminism involved
in environment splitting by introducing X, which is used to check whether subexpressions
do not use the same (linear) variable more than once. We present the algorithm in full and 28 A. Igarashi et al. Fig. 12. Cast insertion. Fig. 12. Cast insertion. Fig. 12. Cast insertion. prove its correctness in Appendix 6. In particular, the algorithm is shown to compute, for
given and e, a minimal type with respect to negative subtyping (if a typing exists). 4 Results We study some of the basic properties (Siek et al., 2015b) of Gradual GV in this section. They include (1) type safety of GGVi and (2) blame safety of GGVi, (3) conservative typing
of GGVe over GV, and (4) the gradual guarantee for GGVe. Since GGVi do not guarantee
deadlock freedom, type safety is stated as the combination of preservation and absence of
runtime errors, rather than progress. We show that (1)–(3) hold with their proof sketches. For (4), we show that GGVe does not satisfy the gradual guarantee. 3.2.5 Cast-inserting translation A well-typed GGVe expression is translated to a GGVi expression by dropping type anno-
tations and inserting casts. Figure 12 presents cast insertion. The judgment ⊢e ⇝f : T
means that “under type environment , a GGVe expression e is translated to a GGVi expres-
sion f at type T.” Most rules are straightforward: casts are inserted where the matching
or consistent subtyping is used. In each rule, blame label p is supposed to be fresh and
positive. The notation f : T
p⇒? U is used to avoid inserting unnecessary casts. f : T
p⇒? U =
f
if T <: U
f : T
p⇒U
otherwise if T <: U Thanks to this optimization, we can show that a program that does not use ⋆or ⋆⃝is
translated to a cast-free GGVi expression, whose behavior obviously coincides with GV. l
d i Thanks to this optimization, we can show that a program that does not use ⋆or ⋆⃝is
translated to a cast-free GGVi expression, whose behavior obviously coincides with GV. For example, we can derive For example, we can derive For example, we can derive ⊢λuno : ⋆.λunc : ⋆⃝.close (send o c)
⇝λuno.λunc.close ((send o (c: ⋆⃝
p⇒!⋆. ⋆⃝)): ⋆⃝
q⇒end!) : ⋆→un ⋆⃝→un unit ⊢λuno : ⋆.λunc : ⋆⃝.close (send o c) un
un
⃝
(
)
⇝λuno.λunc.close ((send o (c: ⋆⃝
p⇒!⋆. ⋆⃝)): ⋆⃝
q⇒end!) : ⋆→un ⋆⃝→un unit ⇝λuno.λunc.close ((send o (c: ⋆⃝
p⇒!⋆. ⋆⃝)): ⋆⃝
q⇒end!) : ⋆→un ⋆⃝→un unit for some p and q. for some p and q. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 29 Gradual session types Fig. 13. Embedding of the unityped calculus. 3.2.6 Embedding One desideratum for a gradual typing system—if it is equipped with dynamic typing—
is that it is possible to embed an untyped (or rather, unityped) language within it (Siek
et al., 2015b). An embedding of an untyped variant of GV into GGVi is given in Figure 13. Blame labels are omitted; each cast should receive a unique blame label. The untyped
variant has the same syntax as the expressions of GV, but every expression has type ⋆and
multiplicities are implicitly assumed to be un. The embedding extends that of (Wadler &
Findler, 2009) for the untyped lambda calculus into the blame calculus. Proof. By case analysis on the last rule used to derive ⊢v : T. Lemma 6 (Substitution). If 1 ⊢v : U and 2, x : U ⊢e : T and = 1 ◦2, then ⊢
e[v/x] : T. Lemma 6 (Substitution). If 1 ⊢v : U and 2, x : U ⊢e : T and = 1 ◦2, then ⊢
e[v/x] : T. Proof. By induction on 2, x : U ⊢e : T with case analysis on the last derivation rule used. We show main cases as follows: Proof. By induction on 2, x : U ⊢e : T with case analysis on the last derivation rule used. We show main cases as follows: Case (variables): If e = x and T = U and un(2), then we have, by Lemma 4, = 1,
finishing the case. If e = y ̸= x, then Lemma 2 finishes the case. Case (variables): If e = x and T = U and un(2), then we have, by Lemma 4, = 1
finishing the case. If e = y ̸= x, then Lemma 2 finishes the case. g
y ̸
Case (applications): We have e = e1 e2 and 11 ⊢e1 : T2 →m T and 12 ⊢e2 : T2 and , x : Case (applications): We have e = e1 e2 and 11 ⊢e1 : T2 →m T and 12 ⊢e2 : T2 and , x U = 11 ◦12. We have two subcases depending on whether un(U) or not. Subcase un(U): We have 11 = ′
11, x : U and 12 = ′
12, x : U and = ′
11 ◦′
12. The induction hypothesis give us ′
11 ◦2 ⊢e1[v/x] : T2 →m T and ′
12 ◦2 ⊢
e2[v/x] : T2. By Lemma 5, we have un(2). The typing rule for applications shows
(′
11 ◦2) ◦(′
12 ◦2) ⊢(e1 e2)[v/x] : T. Lemma 4 finishes the subcase. Subcase lin(U): either (1) 11 = ′
11 and 12 = ′
12, x : U and = ′
11 ◦′
12, in
which case we have ′
12 ⊢e2[v/x] : T1 by the induction hypothesis and also e1[v/x] =
e1 and the typing rule for applications finishes; or (2) 11 = ′
11, x : U and 12 = ′
12
and = ′
11 ◦′
12, in which case the conclusion is similarly proved. □ 4.1 Preservation and absence of runtime errors for GGVi We show preservation and absence of runtime errors for GGVi. The basic structure of the
proof follows Gay and Vasconcelos (2010). In proofs, we often use inversion properties
for the typing relation, such as “if ⊢x: T, then = ′, x: S for some S and ′ such
that S <: T and un(′),” without even stating. They are easy (but tedious) to state and
prove because the only rule that makes typing rules not syntax-directed is T-SUB (see, for
example, (Pierce, 2002) for details). Similarly, we omit inversion for subtyping, which is
syntax-directed. 30 A. Igarashi et al. Lemma 1 (Weakening). If ⊢e: T and un(U), then , x: U ⊢e: T. Proof. By induction on ⊢e: T. □
Lemma 2 (Strengthening). If , x: U ⊢e: T and x does not occur free in e, then ⊢e: T. Proof. By induction on , x: U ⊢e: T. □
Lemma 3 (Preservation for ≡). If P ≡Q, then ⊢P if and only if ⊢Q. Proof. By induction on P ≡Q. Use Lemmas 1, 2, and basic properties of context
splitting (Vasconcelos, 2012; Walker, 2005) for the scope extrusion rules. □
Lemma 4. If = 1 ◦2 and un(1), then = 2. Proof. By induction on = 1 ◦2. □ □ Lemma 4. If = 1 ◦2 and un(1), then = 2. Proof. By induction on = 1 ◦2. □ Lemma 5. If ⊢v : T and un(T), then un(). Proof. By case analysis on the last rule used to derive ⊢v : T. □ □ The following two lemmas are adapted from earlier work (Gay & Vasconcelos, 2010). Lemma 7 (Sub-derivation introduction). If D is a derivation of ⊢E[e] : T, then there
exist 1, 2 and U such that = 1 ◦2 and D has a sub-derivation D′ concluding
2 ⊢e : U and the position of D′ in D corresponds to the position of the hole in E. Lemma 7 (Sub-derivation introduction). If D is a derivation of ⊢E[e] : T, then there
exist 1, 2 and U such that = 1 ◦2 and D has a sub-derivation D′ concluding
2 ⊢e : U and the position of D′ in D corresponds to the position of the hole in E. Lemma 7 (Sub-derivation introduction). If D is a derivation of ⊢E[e] : T, then there
exist 1, 2 and U such that = 1 ◦2 and D has a sub-derivation D′ concluding
2 ⊢e : U and the position of D′ in D corresponds to the position of the hole in E. □ Proof. By induction on E. Proof. By induction on E. 31 Gradual session types Lemma 8 (Sub-derivation elimination). ⊢E[ f ] : T holds, if Lemma 8 (Sub-derivation elimination). ⊢E[ f ] : T holds, if • D is a derivation of 1 ◦2 ⊢E[e] : T, • D′ is a sub-derivation of D concluding 2 ⊢e : U, • the position of D′ in D corresponds to the position of the hole in E, • 3 ⊢f : U, and • = 1 ◦3. • = 1 ◦3. □ Proof. By induction on E. Lemma 9. If ⊢e : T, then flv() = flv(e). Lemma 9. If ⊢e : T, then flv() = flv(e). □ Theorem 1 (Preservation for expressions). If e −→f and ⊢e : T, then ⊢f : T. Theorem 1 (Preservation for expressions). If e −→f and ⊢e : T, then ⊢f : T. Proof. By rule induction on the first hypothesis. For β-reduction and let we use the
substitution lemma (Lemma 6) and inversion of the typing relation. □ Theorem 2 (Preservation for processes). If P −→Q and ⊢P, then ⊢Q. Theorem 2 (Preservation for processes). If P −→Q and ⊢P, then ⊢Q. Proof. By rule induction on the first hypothesis, using basic properties of context split-
ting (Walker, 2005; Vasconcelos, 2012) and weakening (Lemma 1). Rules that make use of
context use sub-derivation introduction (Lemma 7) to build the derivation for the hypoth-
esis, and sub-derivation elimination (Lemma 8) to build the derivation for the conclusion. Rules for reduction to blame use Lemma 9. Reduction underneath parallel composition
and scope restriction follow by induction. The rule for ≡uses Lemma 3. Closure under
evaluation contexts uses Theorem 1. □ Lemma 10 (Ground types, subtyping, and consistent subtyping). Lemma 10 (Ground types, subtyping, and consistent subtyping). 1. If T ̸= ⋆, there is a unique ground type T such that T ∼T. 2. If S ̸= ⋆⃝, there is a unique ground session type S such that S ∼S. 3. T ≲U iff T <: U. 4. S ≲R iff S <: R. Proof. Proof. 1. By case analysis on T. 2. By case analysis on S. 3. By case analysis on T and U. □ 4. By case analysis on S and R. Lemma 11 (Canonical forms). Suppose that ⊢v : T where contains session types and
⋆, only. 1. If T = ⋆, then either v = w: T
p⇒⋆with un(T) or v = a. p
p 1. If T = ⋆, then either v = w: T
p⇒⋆with un(T) or v = a. p 2. If T = S, then either v = c or v = w: S
p⇒⋆⃝and S = ⋆⃝or v = w: R1
p⇒R2 with
R2 <: S. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. Proof. By induction on E. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 32 A. Igarashi et al. 3. If T = unit, then v = (). 3. If T = unit, then v = (). 4. If T = U1 →m U2, then either v = λnx.e with n <: m or v = w: T1 →n1 T2
p⇒
U′
1 →n2 U′
2 with n2 <: m and U1 <: U′
1 and U′
2 <: U2. 4. If T = U1 →m U2, then either v = λnx.e with n <: m or v = w: T1 →n1 T2
p⇒
U′
1 →n2 U′
2 with n2 <: m and U1 <: U′
1 and U′
2 <: U2. 1
2
2
1
2
5. If T = T1 ×m T2, then v = (w1, w2)n with n <: m. 5. If T = T1 ×m T2, then v = (w1, w2)n with n <: m. Proof. By induction on expressions, using Canonical forms (Lemma 11).
□ Proof. By induction on expressions, using Canonical forms (Lemma 11). □ The notion of runtime errors helps us state our type safety result. The subject of an
expression e, denoted by subj(e), is c when e falls into one of the following cases and
undefined in all other cases. send f c
receive c
select l c
case c of {li : xi.fi}i∈I
close c
wait c Proof. By induction on the derivation on ⊢v : T. □ Theorem 3 (Progress for expressions). Suppose that ⊢e : T and that only contains
channel endpoints and references. Then exactly one of the following cases holds: Theorem 3 (Progress for expressions). Suppose that ⊢e : T and that only contains
channel endpoints and references. Then exactly one of the following cases holds: Theorem 3 (Progress for expressions). Suppose that ⊢e : T and that only contains
channel endpoints and references. Then exactly one of the following cases holds: 1. e is a value, 1. e is a value, 2. e −→f (as an expression), 3. e = E[ f ] and f is one of the GV operations: fork f ′, new, send v c, receive c,
select l c, case c of{li : xi.ei}, close c, or wait c, 4. e = E[ f ] and f is a Gradual GV operation: 4. e = E[ f ] and f is a Gradual GV operation: 4. e = E[ f ] and f is a Gradual GV operation: • w: T
p⇒⋆, with lin(T),
• a: ⋆
p⇒U,
• (v : T
p⇒⋆): ⋆
q⇒U, with un(T),
• (v : S
p⇒⋆⃝): ⋆⃝
q⇒R, or
• c: ⋆⃝
p⇒S. • w: T
p⇒⋆, with lin(T),
• a: ⋆
p⇒U,
• (v : T
p⇒⋆): ⋆
q⇒U, with un(T),
• (v : S
p⇒⋆⃝): ⋆⃝
q⇒R, or
• c: ⋆⃝
p⇒S • w: T
p⇒⋆, with lin(T), • w: T
p⇒⋆, with lin(T),
• a: ⋆
p⇒U,
• (v : T
p⇒⋆): ⋆
q⇒U, with un(T),
• (v : S
p⇒⋆⃝): ⋆⃝
q⇒R, or
• c: ⋆⃝
p⇒S • a: ⋆
p⇒U,
p (
)
• c: ⋆⃝
p⇒S. • c: ⋆⃝
p⇒S. □ send f c
receive c d f c
receive c
select l c
case c of {li : xi.fi}i∈I
close c
wait c Two expressions e and f agree on a channel with ends in set {c, d} where c ̸= d, denoted
agree{c,d}{e, f }, a relation on two two-element sets, in the following cases: 1. agree{c,d}{send v c, receive d}; 2. agree{c,d}{select lj c, case d of {li : xi.fi}i∈I} and j ∈I; 3. agree{c,d}{close c, wait d}. 3. agree{c,d}{close c, wait d}. A process is an error if it is structurally congruent to some process that contains a
subprocess of one of the following forms: 1. ⟨E[ve]⟩and v is not an abstraction; 2. ⟨E[let a, b = v in e]⟩and v is not a pair; 3. ⟨E[e]⟩| ⟨F[ f ]⟩and subj(e) = subj( f ); 4. (νc, d)(⟨E[e]⟩| ⟨F[ f ]⟩) and subj(e) = c and subj( f ) = d and not agree{c,d}{e, f The first two cases are typical of functional languages. The third case ensures no two
threads hold references to the same channel endpoint. The fourth case ensures channel
endpoints agree at all times: if one process is ready to send then the other is ready to
receive, and similarly for select and case, close and wait. 33 Gradual session types For processes, rather than a progress result, we present a type safety result as our type
system does not rule out deadlocks, which are formed by a series of processes each waiting
for the next in a circular arrangement; these are exactly the deadlocked processes of GV. Our result holds both for GV and Gradual GV alike. The condition on in the statement
is to exclude processes getting stuck due to a free variable in an application (xe) or a pair
destruction (let a, b = x in e). Theorem 4 (Absence of runtime errors). Let ⊢P where does not contain function or
pair types, and let P −→∗Q. Then Q is not an error. Proof. By induction on the length of reduction steps P −→∗Q. For the base case, where
P = Q, we show P is not an error by showing all error processes cannot be well typed. All cases use Lemma 7 and inversion of the typing relation. The cases for application
and let follow from the fact that does not contain function or pair types. send f c
receive c The third case
follows from the fact that c, being the subject of expressions, is of a linear type, hence
cannot occur in two distinct processes. The fourth case follows from the fact that typability
implies that c and d are of dual types, which in turn implies agree{c,d}(e, f ). □ 4.2 Blame safety Following Wadler & Findler (2009), we introduce three new subtyping relations: <:+,
<:−, and ⊑, called positive, negative, and naive subtyping (also known as precision),
respectively, in Figure 14, in addition to the ordinary subtyping <: defined in Figure 4. A cast from T to U with label p may either return a value or may raise blame labeled
p (called positive blame) or p (called negative blame). The original subtyping relation
T <: U of GGVi characterizes when a cast from T to U never yields blame; relations
T <:+ U and T <:−U characterize when a cast from T to U cannot yield positive or nega-
tive blame, respectively; and relation T ⊑U characterizes when type T is more precise (in
the sense of being less dynamic) than type U. All four relations are reflexive and transitive,
and subtyping, positive subtyping, and naive subtyping are antisymmetric. Wadler & Findler (2009) have an additional rule that makes any subtype of a ground
type a subtype of ⋆, i.e., T <: ⋆if T <: T. This rule is not sound in Gradual GV because
our collide rule blames both casts: ⟨E[(v : T
p⇒⋆): ⋆
q⇒U]⟩−→blame p q (flv(E) ∪flv(v))
if T ̸<: U ⟨E[(v : T
p⇒⋆): ⋆
q⇒U]⟩−→blame p q (flv(E) ∪flv(v))
if T ̸<: U The four subtyping relations are closely related. In previous work (Wadler & Findler,
2009; Siek et al., 2015a) one has that proper subtyping decomposes into positive and neg-
ative subtyping, which—after reversing the order on negative subtyping—recompose into
naive subtyping. Here we have three-quarters of the previous result. Theorem 5 (3/4 Tangram). Theorem 5 (3/4 Tangram). 1. T <: U implies T <:+ U and T <:−U. 2. S <: R implies S <:+ R and S <:−R 3. T ⊑U if and only if T <:+ U and U <:−T. 4. S ⊑R if and only if S <:+ R and R <:−S https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Camb 34
A. Igarashi et al. Fig. 14. Subtyping and blame safety. A. Igarashi et al. 34 Fig. 14. Subtyping and blame safety. Fig. 14. Subtyping and blame safety. Proof. By induction on types. □ Proof. By induction on types. □ □ Proof. By induction on types. Proof. By induction on types. Here the first and second items are an implication, rather than an equivalence as in the
third and fourth items and previous work. In order to get an equivalence, we would need
to alter subtyping such that T <: ⋆for all T and S <: ⋆⃝for all S, which would interfere
with our Canonical Forms lemma (Lemma 11). However, implication in all four items is
sufficient to ensure the most important result, Corollary 1. The definitions of negative subtyping and naive subtyping have been changed since the
conference version of the paper. Now, negative subtyping supports width subtyping and
naive does not. This change is motivated by the type system for the external language, in
particular the join operation. (See the discussion on the join in Section 3.2.4.) The following technical result is used in the proof of Theorem 6. The following technical result is used in the proof of Theorem 6. Lemma 12. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 1. If T ̸= ⋆and T ∼T, then T <:+ T. Lemma 12. 1. If T ̸= ⋆and T ∼T, then T <:+ T. 1. If T ̸= ⋆and T ∼T, then T <:+ T. 2. If S ̸= ⋆⃝and S ∼S, then S <:+ S. 35 Gradual session types Proof. (1) A case analysis on T. Lemma 10 tells us that T is unique. We show the case
for functions. Let T be the type U →m V; we know that T is ⋆→m ⋆, that ⋆<:−U, and
V <:+ ⋆. Conclude with the positive subtyping rule for functions. (2) Similar. □ We say that a process P is safe for blame label p, if all occurrences of casts involving
p or p correspond to subsumptions in the (positive or negative) blame subtyping relation. Figure 14 defines judgments e safe for p and P safe for p, extended homomorphically
to all other forms of expressions and processes. The safe for predicate on well-typed
programs is preserved by reduction. Theorem 6 (Preservation of safe terms). If ⊢P with P safe for p and P −→Q, then
Q safe for p. Proof. It is sufficient to examine all reductions whose contractum involves coercions. We
start with the reductions in Figure 6. The four rules starting from the one with reductum
v : T
p⇒⋆follow from Lemma 12. Then, the standard function cast is analogous to previous
work (Wadler & Findler, 2009), and the case for pairs is similar. The casts for session types
(send, receive, select, case, close, and wait) are new; we concentrate on send. send v (w: !T.S
p⇒!T′.S′) −→(send (v : T′
p⇒T) w): S
p⇒S′ By assumption (w: !T.S
p⇒!T′.S′) safe for p. Inversion of the safe for relation yields
T′ <:∓T and S <:± S′. Hence (v : T′
p⇒T) safe for p and (. . . ): S
p⇒S′ safe for p. Finally, all rules in Figure 7 preserve casts. □ A process P blames label p if P ≡
(Q | R) where Q is blame p q X, blame q p X,
or blame p X, for some q and X, and prefix
of bindings for channel endpoints and
references. Theorem 7 (Progress of safe terms). If ⊢P and P safe for p, then P ̸−→Q where Q
blames p. Proof. We analyze all reduction rules whose contractum includes blame. From Figure 6
take the rule with reductum (v : T
p⇒⋆): ⋆
q⇒U. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Camb https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at A. Igarashi et al. Wadler and Findler (2009) explain how casting between terms related by naive sub-
typing always places the blame (if any) on the less-precisely typed term or context, as
appropriate. Proposition 1 (Consistent Subtyping). Proposition 1 (Consistent Subtyping). 1. T1 ≲T2 if and only if T1 ∼T′
1 and T′
1 <: T2 for some T′
1. 2. T1 ≲T2 if and only if T1 <: T′
2 and T′
2 ∼T2 for some T′
2. 1. T1 ≲T2 if and only if T1 ∼T′
1 and T′
1 <: T2 for some T′
1. 1. T1 ≲T2 if and only if T1
T1 and T1 <: T2 for some T1. 2. T1 ≲T2 if and only if T1 <: T′
2 and T′
2 ∼T2 for some T′
2. 2. T1 ≲T2 if and only if T1 <: T′
2 and T′
2 ∼T2 for some T′
2. 2. T1 ≲T2 if and only if T1 <: T′
2 and T′
2 ∼T2 for some T′
2. Proof. The left-to-right direction is proved by induction on T1 ≲T2 and the right-to-left is
by induction on subtyping with case analysis on T1, T′
2, and T2. □ Proof. The left-to-right direction is proved by induction on T1 ≲T2 and the right-to-left is
by induction on subtyping with case analysis on T1, T′
2, and T2. □ The next lemma clarifies the relation between subtyping, positive and negative subtyp-
ing, and consistent subtyping. Lemma 13 (Subtyping Hierarchy). 4.3 Properties of GGVe Now we turn our attention to GGVe and prove that cast insertion succeeds for well-typed
GGVe expressions and preserves typing and that the GGVe typing conservatively extends
the GV typing. As we need to relate the judgments of different systems, let ⊢e denote the
GGVe typing, ⊢i denote the GGVi typing, and ⊢GV denote the GV typing. Proposition 1 goes back to an observation by Siek and Taha (2007). Lemma 12. It may blame p and q, if T ̸<: U. However,
if it is safe for p then T <:−⋆, which cannot hold (because only ⋆<:−⋆and T cannot be ⋆),
and similar reasoning applies for q and U. The remaining rules are similar. □ We are finally in a position to state the main result of this section. Corollary 1 (Well-typed programs cannot be blamed). Let P be a well-typed process with
a subterm of the form e: T
p⇒U containing the only occurrence of p and p in P. Then the
following cases holds:: • If T <:+ U then P ̸−→∗Q where Q blames p. • If T <:−U then P ̸−→∗Q where Q blames p. • If T <: U then P ̸−→∗Q where Q blames p or p. • If T <: U then P ̸−→∗Q where Q blames p or p. For example, the redex (v : T
p⇒⋆): ⋆
q⇒U may fail and blame p and q if T ̸<: U. And
indeed we have that T ̸<:−⋆and ⋆̸<:+ U, so it is not safe for p or q. However, T <:+ ⋆and For example, the redex (v : T
p⇒⋆): ⋆
q⇒U may fail and blame p and q if T ̸<: U. And
indeed we have that T ̸<:−⋆and ⋆̸<:+ U, so it is not safe for p or q. However, T <:+ ⋆and
⋆<:−U, and the redex will not blame p or q. indeed we have that T ̸<: ⋆and ⋆̸<: U, so it is not safe for p or q. However, T <: ⋆and
⋆<:−U, and the redex will not blame p or q. ⋆<:−U, and the redex will not blame p or q. ⋆<:−U, and the redex will not blame p or q. 36 bridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
m https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core term Lemma 13 (Subtyping Hierarchy). 1. <: ⊆<:+ ⊆≲. 1. <: ⊆<:+ ⊆≲. 2. <: ⊆<:−⊆≲. 2. <: ⊆<:−⊆≲. Proof. <: ⊆<:+ and <: ⊆<:−follow from Theorem 5. <:+ ⊆≲and <:−⊆≲are
by induction on T1 <:+ T2 and T1 <:−T2, respectively. □ Lemma 14 (Upper bound and lower bound). Lemma 14 (Upper bound and lower bound). 1. If T1 ∨T2 = U, then T1 <:−U and T2 <:−U. 2. If T1 ∧T2 = U, then U <:+ T1 and U <:+ T2. 1. If T1 ∨T2 = U, then T1 <:−U and T2 <:−U. 2. If T1 ∧T2 = U, then U <:+ T1 and U <:+ T2. Proof. By simultaneous induction on T1 ∨T2 = U (for the first item) and T1 ∧T2 = U (for
the second item). □ Proof. By simultaneous induction on T1 ∨T2 = U (for the first item) and T1 ∧T2 = U (for
the second item). □ mma 15 (Least upper bound and greatest lower bound). 1. If T1 <:−U and T2 <:−U, then there exists some U′ such that T1 ∨T2 = U′ and
U′ <:−U. 2. If U <:+ T1 and U <:+ T2, then there exists some U′ such that T1 ∧T2 = U′ and
U
:+ U′ 1. If T1 <:−U and T2 <:−U, then there exists some U′ such that T1 ∨T2 = U′ and
U′ <:−U. 2. If U <:+ T1 and U <:+ T2, then there exists some U′ such that T1 ∧T2 = U′ and 1. If T1 <:−U and T2 <:−U, then there exists some U′ such that T1 ∨T2 = U′ and
U′ <:−U. 2. If U <:+ T1 and U <:+ T2, then there exists some U′ such that T1 ∧T2 = U′ and
U <:+ U′. Proof. The two items are simultaneously proved by induction on T1 <:−U and U <:+ T1. □ Theorem 8 states that cast insertion succeeds for well-typed external language and
preserves typing. A few lemmas are required in preparation. Theorem 8 states that cast insertion succeeds for well-typed external language and
preserves typing. A few lemmas are required in preparation. 37 Gradual session types Lemma 16. If T1 ∨T2 = U, then T1 ≲U and T2 ≲U. Lemma 16. If T1 ∨T2 = U, then T1 ≲U and T2 ≲U. Proof. Immediate from Lemmas 13 (1) and 14 (1). □ Lemma 17. If T ▷U, then T ≲U. Lemma 17. Lemma 13 (Subtyping Hierarchy). If T ▷U, then T ≲U. Lemma 17. If T ▷U, then T ≲U. Proof. By case analysis on T ▷U. □ Proof. By case analysis on T ▷U. □ □ Theorem 8 (Cast insertion succeeds and preserves typing). If ⊢e e : T, then there exists
some f such that ⊢e ⇝f : T and ⊢i f : T. Proof. By rules induction on the derivation of ⊢e e : T. We show main cases as follows. Proof. By rules induction on the derivation of ⊢e e : T. We show main cases as follows. Case application rule: We are given Case application rule: We are given Case application rule: We are given = 1 ◦2
e = e1 e2
T = T12
1 ⊢e1 : T1
2 ⊢e2 : T2
T1 ▷T11 →m T12
T2 ≲T11 By 1 ⊢e1 : T1 and the IH, 1 ⊢e1 ⇝f1 : T1 and 1 ⊢f1 : T1 for some f1. By 2 ⊢e2 : T2
and the IH, 2 ⊢e2 ⇝f2 : T2 and 2 ⊢f2 : T2 for some f2. Let By 1 ⊢e1 : T1 and the IH, 1 ⊢e1 ⇝f1 : T1 and 1 ⊢f1 : T1 for some f1. By 2 ⊢e2 : T2
and the IH, 2 ⊢e2 ⇝f2 : T2 and 2 ⊢f2 : T2 for some f2. Let f = ( f1 : T1
p⇒? T11 →m T12) ( f2 : T2
p⇒? T11) By the application rule, ⊢e ⇝f : T. p By the application rule, ⊢e ⇝f : T. p Let us assume f1 : T1
p⇒? T11 →m T12 equals f1 : T1
p⇒T11 →m T12. (If f1 : T1
p⇒? T11 →m
T12 equals f1, we could replace the cast rule with the subsumption rule in what follows.)
We also take similar assumptions in other cases. By T1 ▷T11 →m T12 and Lemma 17, T1 ≲T11 →m T12. By 1 ⊢f1 : T1 and the cast rule, By T1 ▷T11 →m T12 and Lemma 17, T1 ≲T11 →m T12. By 1 ⊢f1 : T1 and the cast rule,
1 ⊢( f1 : T1
p⇒T11 →T12) : T11 →T12 By T1 ▷T11 →m T12 and Lemma 17, T1 ≲T11 →m T12. We say that a type, a type environment, or an expression is static in the following se • A type T is static if T does not contain any dynamic types: i.e., ⋆or ⋆⃝. • A type environment is static if contains only static types. • An expression e of GGVe is static if all types declared in e are static. • An expression e of GGVe is static if all types declared in e are static. Lemma 13 (Subtyping Hierarchy). By 1 ⊢f1 : T1 and the cast rule, 1 ⊢( f1 : T1
p⇒T11 →m T12) : T11 →m T12 f2 : T2 and T2 ≲T11 and the cast rule, By 2 ⊢f2 : T2 and T2 ≲T11 and the cast rule, 2 ⊢( f2 : T2
p⇒T11) : T11 Thus, by the application rule, ⊢f : T. Thus, by the application rule, ⊢f : T. Case case rule: We are given Case case rule: We are given = ′ ◦
e = case e′ of {lj : xj. ej}j∈J
T = U
′ ⊢e′ : T′
T′ ▷&{lj : Rj}j∈J(, xj : Rj ⊢ej : Uj)j∈J
U =
{Uj}j∈J By ′ ⊢e′ : T′ and the IH, ′ ⊢e′ ⇝f : T′ and ′ ⊢f ′ : T′ for some f ′. We take some j ∈
J. By , xj : Rj ⊢ej : Uj and the IH, we have , xj : Rj ⊢ej ⇝fj : Uj and , xj : Rj ⊢fj : Uj
for some fj. Let f = case ( f ′ : T
p⇒? &{lj : Rj}j∈J) of {lj : xj. fj : Uj
p⇒? U}j∈J
le, ⊢e ⇝f : T. f = case ( f ′ : T
p⇒? &{lj : Rj}j∈J) of {lj : xj. fj : Uj
p⇒? U}j∈J By the case rule, ⊢e ⇝f : T. By the case rule, ⊢e ⇝f : T. Next, by T′ ▷&{lj : Rj}j∈J and Lemma 17, T′ ≲&{lj : Rj}j∈J. By ′ ⊢f ′ : T′ and the cast
rule, Next, by T′ ▷&{lj : Rj}j∈J and Lemma 17, T′ ≲&{lj : Rj}j∈J. By ′ ⊢f ′ : T′ and the cast
rule, Next, by T′ ▷&{lj : Rj}j∈J and Lemma 17, T′ ≲&{lj : Rj}j∈J. By ′ ⊢f ′ : T′ and the cast
rule, https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 38 A. Igarashi et al. Lemma 18. 1. Suppose T, U are static. If T ≲U, then T <: U. 2. Suppose T ▷U and T ̸= ⋆, ⋆⃝. a. If U is neither &-type nor ⊕-type, then T = U. b. If U is either &-type or ⊕-type, then T <: U. c. If T is static and U is not &-type, then U is static. 3. Suppose T1, T2 are static. If T1 ∨T2 = U, then a. U is static,
b. T1 <: U and T2 <: U,
c. U <: U′ for any static U′ such that T1 <: U′ and T2 <: U′. a. U is static, a. U is static,
b. T1 <: U and T2 <: U, b. T1 <: U and T2 <: U, c. U <: U′ for any static U′ such that T1 <: U′ and T2 <: U′. 4. Suppose T1, T2, U′ are static. If T1 <: U′ and T2 <: U′, then there exists some static
U such that U = T1 ∨T2. 4. Suppose T1, T2, U′ are static. If T1 <: U′ and T2 <: U′, then there exists some static
U such that U = T1 ∨T2. Proof. The first item is by induction on T ≲U. The second item is by case analysis on
T ▷U. Here, we can prove Proof. The first item is by induction on T ≲U. The second item is by case analysis on
T ▷U. Here, we can prove if T, U are static and T <:−U, then T <: U if T, U are static and T <:−U, then T <: U if T, U are static and T <:−U, then T <: U by induction on T <:−U. By Lemma 13, <: ⊆<:−. Thus, we have by induction on T <:−U. By Lemma 13, <: ⊆<:−. Thus, we have if T, U are static, then T <: U if and only if T <:−U. if T, U are static, then T <: U if and only if T <:−U. With this fact, the third and fourth items can be proved by Lemmas 14 and 15, respectively
□ With this fact, the third and fourth items can be proved by Lemmas 14 and 15, respectively. □ We define the type erasure |e|, which is obtained by removing type annotations from an
expression e of GGVe. ′ ⊢( f ′ : T′
p⇒&{lj : Rj}j∈J) : &{lj : Rj}j∈J ′ ⊢( f ′ : T′
p⇒&{lj : Rj}j∈J) : &{lj : Rj}j∈J
J. By U = {Uj}j∈J and Lemma 16, Uj ≲U. By , xj : Rj ⊢fj : Uj and ( f
⇒&{ j
j}j∈J) &{ j
j}j∈J
We take some j ∈J. By U = {Uj}j∈J and Lemma 16, Uj ≲U. By , xj : Rj ⊢fj : Uj and
the cast rule, ( f
{ j
j}j
)
{ j
j}j
We take some j ∈J. By U = {Uj}j∈J and Lemma 16, Uj ≲U. By , xj : Rj ⊢fj : Uj and
the cast rule, , xj : Rj ⊢( fj : Uj
p⇒U) : U , xj : Rj ⊢( fj : Uj
p⇒U) : U □ us, by the case rule, ⊢f : T. □ Thus, by the case rule, ⊢f : T. Thus, by the case rule, ⊢f : T. □ We say that a type, a type environment, or an expression is static in the following sense: Case application rule: We are given = 1 ◦2
e = e1 e2
T = T12
1 ⊢e1 : T1
2 ⊢e2 : T2
T1 ▷T11 →m T12
T2 ≲T11 = 1 ◦2
e = e1 e2
T = T12
1 ⊢e1 : T1
2 ⊢e2 : T2
T1 ▷T11 →m T12
T2 ≲T11 Since , e are static, 1, 2, e1, e2 are also static. By 1 ⊢e1 : T1 and the IH, T1 is static and
1 ⊢|e1| : T1. By T1 ▷T11 →m T12 and Lemma 18 (2), T11, T12 are static and T1 = T11 →m
T12. By 2 ⊢e2 : T2 and the IH, T2 is static and 2 ⊢|e2| : T2. By T2 ≲T11 and Lemma
18 (1), T2 <: T11. By the subsumption rule, 2 ⊢|e2| : T11. Thus, by 1 ⊢|e1| : T11 →m T12
2 ⊢|e2| : T11
|e1 e2| = |e1| |e2|
and the application rule, we have 1 ◦2 ⊢|e1 e2| : T12. 1 ⊢|e1| : T11 →m T12
2 ⊢|e2| : T11
|e1 e2| = |e1| |e2|
and the application rule, we have 1 ◦2 ⊢|e1 e2| : T12. Case case rule: We are given = ′ ◦
e = case e′ of {lj : xj. ej}j∈J
T = U
′ ⊢e′ : T′
T′ ▷&{lj : Rj}j∈J
(, xj : Rj ⊢ej : Uj)j∈J
U =
{Uj}j∈J Since , e are static, ′, , e′, ej are also static. Since any type environment , xj : Rj is
static, any Rj is static. By ′ ⊢e′ : T′ and the IH, T′ is static and ′ ⊢|e′| : T′. By T′ ▷&{lj :
Rj}j∈J and Lemma 18 (2), T′ <: &{lj : Rj}j∈J. By the subsumption rule, Since , e are static, ′, , e′, ej are also static. Since any type environment , xj : Rj is
static, any Rj is static. By ′ ⊢e′ : T′ and the IH, T′ is static and ′ ⊢|e′| : T′. By T′ ▷&{lj :
Rj}j∈J and Lemma 18 (2), T′ <: &{lj : Rj}j∈J. By the subsumption rule, ′ ⊢|e′| : &{lj : Rj}j∈J We take some j ∈J. Since , xj : Rj is static, by , xj : Rj ⊢ej : Uj and the IH, Uj is static
and , xj : Rj ⊢|ej| : Uj. Lemma 18. The main cases of its definition are as follows: |new S| = new
|λmx:T. e| = λmx. |e| |new S| = new
|λmx:T. e| = λmx. |e| |new S| = new |λmx:T. e| = λmx. |e| (It is extended homomorphically for all other forms of expressions.) (It is extended homomorphically for all other forms of expressions.) https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Camb 39 Gradual session types Theorem 9 states that the GGVe typing is a conservative extension of the GV typing. We have to take care of the difference between the declarative type system of GV and the
algorithmic type system of GGVe. Theorem 9 (Typing Conservation over GV). Suppose that is static. heorem 9 (Typing Conservation over GV). Suppose that is static. 1. If e is static and type environments that appear in the derivation of ⊢e e : T are
all static, then T is static and ⊢GV |e| : T. 2. If f is an expression of GV and ⊢GV f : T, then T is static and there exist static e
and static T′ such that |e| = f and ⊢e e : T′ and T′ <: T. Proof. The first item is by induction on ⊢e e : T with case analysis on the rule applied
last. We show the main cases as follows. Case application rule: We are given Case application rule: We are given By U = {Uj}j∈J and Lemma 18 (3), Uj <: U and U is static. By
the subsumption rule, , xj : Rj ⊢|ej| : U Thus, by ′ ⊢|e′| : &{lj : Rj}j∈J
(, xj : Rj ⊢|ej| : U)j∈J
|case e′ of {lj : xj. ej}j∈J| = case |e′| of {lj : xj. |ej|}j∈J
case rule, we have ′ ◦ ⊢|case e′ of {lj : xj. ej}j∈J| : U. ′ ⊢|e′| : &{lj : Rj}j∈J
(, xj : Rj ⊢|ej| : U)j∈J
|case e′ of {lj : xj. ej}j∈J| = case |e′| of {lj : xj. |ej|}j∈J the case rule, we have ′ ◦ ⊢|case e′ of {lj : xj. ej}j∈J| : U. the case rule, we have ′ ◦ ⊢|case e′ of {lj : xj. ej}j∈J| : U. The second item is by induction on ⊢GV f : T with case analysis on the rule applied
last. We show the main cases as follows. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at A. Igarashi et al. 40 Case application rule: We are given Case application rule: We are given = 1 ◦2
f = f1 f2
T = T12
1 ⊢f1 : T11 →m T12
2 ⊢f2 : T11 Since is static, 1, 2 are also static. By 1 ⊢f1 : T11 →m T12 and the IH, T11, T12 are
static and there exist static e1, U1 such that Since is static, 1, 2 are also static. By 1 ⊢f1 : T11 →m T12 and the IH, T11, T12 are
static and there exist static e1, U1 such that 1 ⊢e1 : U1
U1 <: T11 →m T12
|e1| = f1 By inversion of <:, we have U1 = U11 →n U12 and n <: m and T11 <: U11 and U12 <: T12
for some U11, U12, n. Since U1 is static, U11, U12 are also static. By 2 ⊢f2 : T11 and the
IH, T11 is static and there exist static e2, U2 such that By inversion of <:, we have U1 = U11 →n U12 and n <: m and T11 <: U11 and U12 <: T12
for some U11, U12, n. Since U1 is static, U11, U12 are also static. Case application rule: We are given By 2 ⊢f2 : T11 and the
IH, T11 is static and there exist static e2, U2 such that 2 ⊢e2 : U2
U2 <: T11
|e2| = f2 By U2 <: T11 and T11 <: U11 and transitivity, U2 <: U11. By Lemma 13, U2 ≲U11. From
Figure 9, we have U11 →n U12 ▷U11 →n U12. Thus, by By U2 <: T11 and T11 <: U11 and transitivity, U2 <: U11. By Lemma 13, U2 ≲U11. From
Figure 9, we have U11 →n U12 ▷U11 →n U12. Thus, by 1 ⊢e1 : U11 →n U12
2 ⊢e2 : U2
U11 →n U12 ▷U11 →n U12
U2 ≲U11 1 ⊢e1 : U11 →n U12
2 ⊢e2 : U2
U11 →n U12 ▷U11 →n U12
U2 ≲U11
and the application rule, 1 ◦2 ⊢e1 e2 : U12. Additionally,
|e1 e2| = |e1| |e2| = f1 f2 = f
U12 <: T12 = T and the application rule, 1 ◦2 ⊢e1 e2 : U12. Additionally, |e1 e2| = |e1| |e2| = f1 f2 = f
U12 <: T12 = T Case case rule: We are given Case case rule: We are given = ′ ◦
f = case f ′ of {lj : xj. fj}j∈J
′ ⊢f ′ : &{lj : Rj}j∈J
(, xj : Rj ⊢fj : T)j∈J Since is static, ′, are also static. By ′ ⊢f ′ : &{lj : Rj}j∈J and the IH, all Rj are static
and there exist static e′, T′ such that Since is static, ′, are also static. By ′ ⊢f ′ : &{lj : Rj}j∈J and the IH, all Rj are static
and there exist static e′, T′ such that |e′| = f ′
′ ⊢e′ : T′
T′ <: &{lj : Rj}j∈J By T′ <: &{lj : Rj}j∈J and Lemma 13, T′ ≲&{lj : Rj}j∈J. We take some j ∈J. By , xj : Rj ⊢
fj : T and the IH, T is static and there exist static ej, Tj such that By T′ <: &{lj : Rj}j∈J and Lemma 13, T′ ≲&{lj : Rj}j∈J. We take some j ∈J. By , xj : Rj ⊢
fj : T and the IH, T is static and there exist static ej, Tj such that |ej| = fj
, xj : Rj ⊢ej : Tj
Tj <: T So, (Tj <: T)j∈J. Case application rule: We are given Case application rule: We are given = 1 ◦2
e = e1 e2
f = ( f1 : T1
p⇒? T11 →m T12) ( f2 : T2
p⇒? T11)
T = T12
1 ⊢e1 ⇝f1 : T1
2 ⊢e2 ⇝f2 : T2
T1 ▷T11 →m T12
T2 ≲T11 Since , e are static, 1, 2, e1, e2 are also static. By 1 ⊢e1 ⇝f1 : T1 and the IH, T1 is
static and |e1| = f1. By T1 ▷T11 →m T12 and Lemma 18 (2), T11, T12 are static and T1 =
T11 →m T12. So, f1 : T1
p⇒? T11 →m T12 = f1 = |e1| f1 : T1
p⇒? T11 →m T12 = f1 = |e1| By 2 ⊢e2 ⇝f2 : T2 and the IH, T2 is static and |e2| = f2. By T2 ≲T11 and Lemma 18 (1),
T2 <: T11. So, T2 <: T11. So, f2 : T2
p⇒? T11 = f2 = |e2| Thus, f = |e1| |e2| = |e1 e2|. □ Thus, f = |e1| |e2| = |e1 e2|. Case application rule: We are given By Lemma 18 (4), there exist some static U such that U = {Tj}j∈J. By
Lemma 18 (3), U <: T. Thus, by So, (Tj <: T)j∈J. By Lemma 18 (4), there exist some static U such that U = {Tj}j∈J. By
Lemma 18 (3), U <: T. Thus, by ( j
)j
y
(
Lemma 18 (3), U <: T. Thus, by ( j
)j
y
( )
Lemma 18 (3), U <: T. Thus, by ′ ⊢e′ : T′
(, xj : Rj ⊢ej : Tj)j∈J
T′ ≲&{lj : Rj}j∈J
U =
{Tj}j∈J and the case rule, ′ ◦ ⊢case e′ of {lj : xj. ej}j∈J : U. Additionally, and the case rule, ′ ◦ ⊢case e′ of {lj : xj. ej}j∈J : U. Additionally, |case e′ of {lj : xj. ej}j∈J| = case |e′| of {lj : xj. |ej|}j∈J = case f ′ of {lj : xj. fj}j∈J = f |case e′ of {lj : xj. ej}j∈J| = case |e′| of {lj : xj. |ej|}j∈J = case f ′ of {lj : xj. fj}j∈J = f
We already have U <: T. Case subsumption rule: We are given ⊢f : U and U <: T. By the IH, U is static and
there exist static e and U′ such that ⊢e : U′
U′ <: U
|e| = f By U′ <: U and U <: T and transitivity, U′ <: T. □ □ By U′ <: U and U <: T and transitivity, U′ <: T. By U′ <: U and U <: T and transitivity, U′ <: T. By U′ <: U and U <: T and transitivity, U′ <: T. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 41 Gradual session types Proposition 2 states that the cast-insertion translation does not insert casts for static
expressions, which can be seen as expressions of GV if type annotations are removed. The
proof is similar to that of Theorem 9 (1). Proposition 2. Suppose that and e are both static. If ⊢e ⇝f : T, then T is static and
|e| = f . Proof. By induction on ⊢e ⇝f : T. with case analysis on the rule applied last. We show
one of the main cases as follows. Using the precision, the static gradual guarantee can be stated as follows. If 1 ⊑2, e1 ⊑e2, and 1 ⊢e1 : T1, then 2 ⊢e2 : T2 and T1 ⊑T2 for some T2. If 1 ⊑2, e1 ⊑e2, and 1 ⊢e1 : T1, then 2 ⊢e2 : T2 and T1 ⊑T2 for some T2. However, it does not hold. Theorem 10 (Failure of the Static Gradual Guarantee). There exist 1, 2, e1, e2, and T1
such that 1 ⊑2, e1 ⊑e2, 1 ⊢e e1 : T1 and, for any T2 such that 2 ⊢e e2 : T2, T1 ̸⊑T2. Proof. Let 1 = x : T1, y : T2, z : &{l1 : end!, l2 : end!}
2 = x : ⋆, y : T2, z : &{l1 : end!, l2 : end!}
e1 = e2 = case z of {l1 : x1. close x1; x, l2 : x2. close x2; y}
T1 = unit →lin unit
T2 = unit →un unit (where e1; e2 stands for usual sequential composition). Then, 1 ⊑2, e1 ⊑e2, 1 ⊢e e1 :
T1, and 2 ⊢e e2 : T2; but T1 ̸⊑T2. □ (where e1; e2 stands for usual sequential composition). Then, 1 ⊑2, e1 ⊑e2, 1 ⊢e e1 :
T1, and 2 ⊢e e2 : T2; but T1 ̸⊑T2. □ Although we do not state the dynamic gradual guarantee formally, we expect at least
that, if two programs e1 and e2 satisfy e1 ⊑e2 and the execution of e1 (after cast insertion)
terminates normally (at ⟨()⟩), then e2 also terminates normally. Unfortunately, it would not
be very difficult to see such an expectation fail. Let’s consider e′
1 = (λlinx:T1. e1) (λlinx1:unit. x1) and a more imprecise expression e′
2 = (λlinx:⋆. e2) (λlinx1:unit. x1) The former will return λlinx1:unit. x1 if l1 is selected by another process. However, e′
2 shows
different behavior: the cast-inserting translation puts a cast from ⋆to T2 = unit →un unit
on x in the first branch of case in e2 but x will be bound to (a reference to) a linear function
and, if l1 is selected, the cast will fail and raise blame. The problem seems to stem from the fact that ∨has subtle interaction with ⊑. For
typing case-expressions, we would naturally require precision to be preserved by the join
operation, i.e., if T1 ⊑T′
1, then T1 ∨T2 ⊑T′
1 ∨T2. 4.4 (Failure of) The gradual guarantee In a gradually typed language, changing type annotations in a program should not change
the static or dynamic behavior—except for runtime errors caused by casts. Such an expec-
tation is formalized by Siek et al. (2015b) as the gradual guarantee property. It usually
consists of two statements concerning the static and dynamic aspects of programs. The
static counterpart of the gradual guarantee (simply called the static gradual guarantee)
states that less precise type annotations make the type of an expression less precise,
whereas the dynamic gradual guarantee states that making type annotations less precise
does not change the final outcome of a program. We will show that, unfortunately, GGVe satisfies neither the static nor dynamic gradual
guarantee by constructing counterexamples. We analyze the problem and argue that it is
not easy to recover without losing other good properties. First, to capture the notion of programs with more precise type annotations formally, the
precision over types is extended to type environments and expressions. The relation 1 ⊑
2 is the least relation that satisfies · ⊑· and 1, x : T1 ⊑2, x : T2 if 1 ⊑2 and T1 ⊑T2
and the relation e1 ⊑e2 is the least precongruence that is closed under the following rules: S1 ⊑S2
new S1 ⊑new S2
(1) T1 ⊑T2
e1 ⊑e2
λmx:T1. e1 ⊑λmx:T2. e2 (1) 42 A. Igarashi et al. 5.1 Gradual typing Findler and Felleisen (2002) introduced two seminal ideas: higher-order contracts that
dynamically monitor conformance to a type discipline, and blame to indicate whether
it is the library or the client which is at fault if the contract is violated. Siek and Taha
(2006, 2007) introduced gradual types to integrate untyped and typed code, while Flanagan
(2006) introduced hybrid types to integrate simple types with refinement types. Both used
target languages with explicit casts and similar translations from source to target; both
exploit contracts, but neither allocates blame. Motivated by similarities between gradual
and hybrid types, Wadler and Findler (2009) introduced blame calculus, which unifies the
two by encompassing untyped, simply typed, and refinement-typed code. As the name indi-
cates, it also restores blame, which enables a proof of blame safety: blame for type errors
always lays with less-precisely typed code—“well-typed programs can’t be blamed.” While the first investigations of gradual typing were based on simply typed calculi,
subsequent work has explored gradual typing for a range of typing features. Polymorphism
(Ahmed et al., 2011; Igarashi et al., 2017b; Toro et al., 2019) has proved to be quite tricky,
with one important question about the Jack-of-All-Trades Principle (Ahmed et al., 2011)
still open. A gradual treatment of record types may be found in the paper on Abstract
Gradual Typing (AGT) (Garcia et al., 2016). Variant types have proved elusive, but union
types have been considered (Siek & Tobin-Hochstadt, 2016) along with intersection types
and polymorphism as part of a set-theoretical reevaluation of gradual principles (Castagna
et al., 2019). Moving toward session types, systems with gradual typestate have been considered
(Wolff et al., 2011; Garcia et al., 2014); they extend an object-oriented language with
typestate by a dynamic type and define a suitable translation to an internal language with
casts. The additional complication is to track the current typestate at runtime. Thiemann
(2014) describes a system with gradual types and session types, but in it only types (and
not session types) can be gradual. Effect systems have been gradualized based on ideas from abstract interpretation by
Banados Schwerter and others (2014). While the former work only presented a gradualiza-
tion of effects themselves a subsequent extension adds a full treatment of types (Schwerter
et al., 2016). Related ideas are explored by Thiemann and Fennell (2014), who present an
approach to gradualize annotated type systems, like units and security labels. Using the precision, the static gradual guarantee can be stated as follows. However, the current definition of ∨
breaks this property as the counterexample to the static gradual guarantee above shows. Also, as we can see from the counterexample to the dynamic gradual guarantee, join with
a more precise type can yield a supertype—that is, T1 ⊑⋆and ⋆∨T2 <: T1 ∨T2 hold. One possible workaround is to adapt the “lifted join” operation ¨∨of the GTFL≲lan-
guage (Garcia et al., 2016) to Gradual GV. Like ∨, unit →lin unit ¨∨unit →un unit =
unit →lin unit but, unlike ∨, ⋆¨∨unit →un unit = ⋆. Thus, the lifted join would perhaps
recover the gradual guarantee. However, it seems that the lifted join is the least upper
bound operation for no known ordering between types and we would lose the minimal
type property of GGVe if we used ¨∨. Also, the lifted join has the following property:
T ¨∨⋆is T only if T does not have nontrivial supertypes; otherwise T ¨∨⋆is ⋆. For
example, unit ¨∨⋆= unit and unit →lin unit ¨∨⋆= unit →lin unit but unit →un unit ¨∨⋆= ⋆ 43 Gradual session types (because unit →un unit <: unit →lin unit). It means that the standard narrowing property—
if , x : T1 ⊢e e : T and T2 <: T1, then 1, x : T2 ⊢e e : T—does not hold. We leave more
detailed analysis of the problem and possible remedy for future work. 5.1 Gradual typing Many of the
above-cited works strive to fulfill the properties of Siek and others, not all of them are
successful, but further discussion of the properties exceeds the scope of this survey of
related work. Siek and others (2015b) review desirable properties of gradually typed languages, while
Wadler Wadler (2015) discusses history of the blame calculus and why blame is important. These papers provide overviews of the field, each with many further citations. Many of the
above-cited works strive to fulfill the properties of Siek and others, not all of them are
successful, but further discussion of the properties exceeds the scope of this survey of
related work. TypeScript TPD (Williams et al., 2017) applies contracts to monitor the gradual typ-
ing of TypeScript, and evaluates the successes and shortcomings of contracts in this
context. 5.1 Gradual typing Following an
earlier approach for gradual security typing for simply typed lambda calculus (Disney &
Flanagan, 2011), Fennell and Thiemann (2013) developed gradual security for an ML core
language with references and subsequently for a Java core language LJGS with polymor-
phic security labels (Fennell & Thiemann, 2016). Toro and others (2018) developed a
gradual calculus with slightly different features from first principles using the AGT (Garcia
et al., 2016) approach. In each of these approaches, special measures have to be taken to 44 A. Igarashi et al. ensure the key property of noninterference. Gradual type systems related to session types
also include the runtime enforcement of affine typing of Tov and Pucella Tov and Pucella
(2010). As noted in the introduction, gradual typing may be important as a bridge to type systems
that go beyond what is currently available, including dependent, effect, and session types. There is a range of gradual type systems for dependent types. Ou and others (2004) bridge
the gap between simply typed lambda calculus and a calculus with indexed types. In
Flanagan’s hybrid typing (2006), subtyping judgments are either proved or disproved stat-
ically by SMT theorem proving or residualized as runtime checks. Greenberg and others
(2010) consider different styles of contracts in simply typed and dependently typed set-
tings. Lehmann and Tanter (2017) present an approach that uses the AGT methodology
to obtain a gradual system that mediates between simple types and dependent refinement
types. This work has been augmented with type inference by Vazou and others (2018) and
it has been extended to verification (Bader et al., 2018) where specifications may contains
unknown subformulas. Jafery & Dunfield (2017) consider gradualized refinements for sum
types with the goal to control errors in pattern matching. Gradual ownership types (Sergey & Clarke, 2012) is a gradualization of the Owners
as Dominators principle of ownership. Its theory is built with similar principles as other
gradual languages, but its flavor is different as ownership is not a semantic property, but a
structure imposed by the programmer. Siek and others (2015b) review desirable properties of gradually typed languages, while
Wadler Wadler (2015) discusses history of the blame calculus and why blame is important. These papers provide overviews of the field, each with many further citations. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 5.2 Session types Session types were introduced by Honda (1993) and Honda et al. (1998). The original
system addressed binary sessions, whereby types describe the interaction between two
partners. Binary sessions were eventually extended to the more general setting of multi-
party session types (Honda et al., 2016). Recent years have seen the introduction of session
types in programming languages, and software development tools. We review the most
important works. Session types inspired the design of several programming languages. Sing# (Fähndrich
et al., 2006) constitutes one of the first attempts to introduce session types in programming
languages. An extension of C, Sing# was used to implement Singularity, an operating sys-
tem based on message passing. Gay and others (2010) propose attaching session types to
class definitions, allowing to treat channels as objects for session-based communication in
distributed systems. SePi (Franco & Vasconcelos, 2013) is a concurrent, message-passing
programming language based on the pi-calculus, featuring a simple form of refinement
types. SILL (Toninho et al., 2013; Pfenning & Griffith, 2015) is a higher-order session 45 Gradual session types functional programming language, featuring process expressions as first class objects via
a linear contextual monad. Concurrent C0 (Willsey et al., 2017) is a type-safe C-like pro-
gramming language equipped with channel communication governed by session types. Links (Lindley & Morris, 2017) is a functional programming language designed for tierless
web applications that natively supports binary session types. Proposals have been made to retroactively introduce session types in mainstream
programming languages. Session Java (Hu et al., 2008) introduces API-based session
primitives in Java, while (Hu et al., 2010) presents a Java language extension and type dis-
cipline for session-based event-driven programming. Featherweight Erlang (Mostrous &
Vasconcelos, 2011) imposes a session-based type system to discipline message passing in
Erlang. Mungo (Kouzapas et al., 2016) is a tool for checking Java code against session
types, presented in the form of typestates. Embedding of session types have been pro-
posed for Haskell (Pucella & Tov, 2008; Sackman & Eisenbach, 2008; Polakow, 2015;
Orchard & Yoshida, 2016; Lindley & Morris, 2016a), OCaml (Padovani, 2017), Scala
(Scalas & Yoshida, 2016), and Rust (Jespersen et al., 2015). Most of these embeddings
delegate linearity checks to the runtime system. 5.2 Session types Session types can be used in the software development process under different forms,
including languages to describe protocols, specialized libraries to invoke session-based
communication primitives, provision for runtime monitoring against session types, and
extended type checkers. Scribble (Honda et al., 2011) is a language-agnostic protocol
description formalism used in many different tools. Multiparty Session C (Ng et al.,
2012) uses Scribble, a compiler plug-in, and a C library to validate against session types. Hu and Yoshida (2016) generate protocol-specific Java APIs from multiparty session
types described in Scribble. SPY (Neykova et al., 2013) generates runtime monitors for
endpoint communication from Scribble protocols. Neykova and Yoshida (2014) designed
and implemented a session actor library in Python together with a runtime verification
mechanism. Bocchi and others (2017) present a theory that incorporates both static typing
and dynamic monitoring of session types. Fowler (2016) describes a framework for
monitoring Erlang applications against multiparty session types. Neykova and Yoshida
(2017) investigate failure handling for Erlang processes in a system that dynamically
monitors session types. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at References Ahmed, A., Findler, R. B., Siek, J. G. & Wadler, P. (2011) Blame for all. In Proceedings of the 38th
ACM SIGPLAN-SIGACT Symposium on Principles of Programming Languages, POPL 2011,
Austin, TX, USA, January 26–28, 2011, Ball, T. & Sagiv, M. (eds), ACM, pp. 201–214. É Bader, J., Aldrich, J. & Tanter, É. (2018) Gradual program verification. In Verification, Model
Checking, and Abstract Interpretation – 19th International Conference, VMCAI 2018, Los
Angeles, CA, USA, January 7–9, 2018, Proceedings, Dillig, I. & Palsberg, J. (eds). Lecture Notes
in Computer Science, vol. 10747. Springer, pp. 25–46. Bañados Schwerter, F., Garcia, R. & Tanter, É. (2014) A theory of gradual effect systems. In
International Conference on Functional Programming (ICFP). ACM, pp. 283–295. Barendregt, H. P. (1984) The Lambda Calculus: Its Syntax and Semantics. North-Holland. Bierman, G., Abadi, M. & Torgersen, M. (2014) Understanding TypeScript. In European Conference
on Object-Oriented Programming (ECOOP). LNCS, vol. 8586. Springer, pp. 257–281. Bierman, G. M., Meijer, E. & Torgersen, M. (2010) Adding dynamic types to C#. In European
Conference on Object-Oriented Programming (ECOOP). LNCS. Springer, pp. 76–100. Bocchi, L., Chen, T.-C., Demangeon, R., Honda, K. & Yoshida, N. (2013) Monitoring networks
through multiparty session types. In Formal Techniques for Distributed Systems, Beyer, D. &
Boreale, M. (eds). Springer, pp. 50–65. Bocchi, L., Chen, T.-C., Demangeon, R., Honda, K. & Yoshida, N. (2017) Monitoring networks
through multiparty session types. Theor. Comput. Sci. 699, 33–58. Brady, E. (2013) Idris, a general-purpose dependently typed programming language: Design an
implementation. J. Funct. Program. 23(05), 552–593. Brady, E. (2013) Idris, a general-purpose dependently typed programming language: Design and
implementation. J. Funct. Program. 23(05), 552–593. Caires, L. & Pfenning, F. (2010) Session types as intuitionistic linear propositions. In International Brady, E. (2013) Idris, a general-purpose dependently typed programming language: Design and
implementation. J. Funct. Program. 23(05), 552–593. Caires, L. & Pfenning, F. (2010) Session types as intuitionistic linear propositions. In International
Conference on Concurrency Theory (CONCUR) LNCS Springer pp 222 236 Caires, L. & Pfenning, F. (2010) Session types as intuitionistic linear propositions. In Internationa
Conference on Concurrency Theory (CONCUR). LNCS. Springer, pp. 222–236. Caires, L., Pfenning, F. & Toninho, B. (2014) Linear logic propositions as session types. Math
Struct. Comput. Sci. 26(03), 367–423. Carbone, M., Honda, K. & Yoshida, N. (2007) Structured communication-centred programming
for web services. In De Nicola, R. Acknowledgments We would like to thank Alceste Scalas and Nobuko Yoshida for comments and point-
ing out errors in the definition of subtyping rules, Kaede Kobayashi for pointing out
subtle errors in the operational semantics, and anonymous reviewers for construc-
tive comments. We are also grateful to Hannes Saffrich for implementing a type
checker for the calculus in this paper. This work was supported in part by the JSPS
KAKENHI Grant Number JP17H01723 (Igarashi), by FCT through the LASIGE Research
Unit ref. UID/CEC/00408/2019 and project Confident ref. PTDC/EEI-CTP/4503/2014
(Vasconcelos), and by EPSRC program grant EP/K034413/1 (Wadler). 6 Conclusions We presented the design of Gradual GV, which combines a session-typed language GV
along the lines of Gay and Vasconcelos (2010) with a blame calculus along the lines of
Wadler and Findler (2009), and with dynamic enforcement of linearity along the lines of
Tov and Pucella (2010). We established expected results for such a language, including
type safety and blame safety. The gradual guarantee however does not hold, and it is not
clear how it can be recovered without losing other good properties. Much remains to be done; we consider just one future direction here. The embed-
ding of linear types in the unrestricted dynamic type relies on an indirection through
a cell in the store. In our present work, these cells are used once and then discarded. This one-shot policy imposes a certain usage pattern on linear values embedded in the
unityped language. In particular, the send and receive operations on a channel need A. Igarashi et al. 46 to be chained as in (close (send v2 (send v1 c))). However, one could imagine a uni-
typed language where one may use the channel nonlinearly in an imperative style as in
(send v1 c; send v2 c; close c), mimicking the style of network programming in conven-
tional languages. This style can also be supported by a variant of Gradual GV with a
multi-shot policy that restores an updated channel to the same cell from which it was
extracted. We leave the full formalization of this policy to future work. References (ed), Programming Languages and Systems, 16th European
Symposium on Programming, ESOP 2007, Held as Part of the Joint European Conferences 47 Gradual session types on Theory and Practics of Software, ETAPS 2007, Braga, Portugal, March 24–April 1, 2007,
Proceedings. Lecture Notes in Computer Science, vol. 4421. Springer, pp. 2–17. Carbone, M., Honda, K. & Yoshida, N. (2012) Structured communication-centered programming for
web services. ACM Trans. Program. Lang. Syst. 34(2), 8. Castagna, G., Lanvin, V., Petrucciani, T. & Siek, J. G. (2019) Gradual typing: A new perspective. PACMPL 3(POPL), 16:1–16:32. Chaudhuri, A., Vekris, P., Goldman, S., Roch, M. & Levi, G. (2017) Fast and precise type checking
for JavaScript. PACMPL 1(OOPSLA), 48:1–48:30. Cimini, M. & Siek, J. G. (2016) The Gradualizer: A methodology and algorithm for generating
gradual type systems. In Proceedings of the 43rd Annual ACM SIGPLAN-SIGACT Symposium
on Principles of Programming Languages, POPL 2016, St. Petersburg, FL, USA, January 20–22,
2016, pp. 443–455. Cooper, E., Lindley, S., Wadler, P. & Yallop, J. (2007) Links: Web programming without tiers. In
Formal Methods for Components and Objects, de Boer, F. S., Bonsangue, M. M., Graf, S. & de
Roever, W. P. (eds). Springer, pp. 266–296. Demangeon, R. & Honda, K. (2011) Full abstraction in a subtyped pi-calculus with linear types. In Katoen, J.-P. & König, B. (eds), CONCUR 2011 - Concurrency Theory - 22nd International
Conference, CONCUR 2011, Aachen, Germany, September 6–9, 2011. Proceedings. Lecture
Notes in Computer Science, vol. 6901. Springer, pp. 280–296. Demangeon, R., Honda, K., Hu, R., Neykova, R. & Yoshida, N. (2015) Practical interruptible con-
versations: Distributed dynamic verification with multiparty session types and python. Formal
Methods Syst. Des. 46(3), 197–225. Disney, T. & Flanagan, C. (2011) Gradual information flow typing. In STOP. Available at:
https://users.soe.ucsc.edu/∼cormac/papers/stop11.pdf Ernst, E., Møller, A., Schwarz, M. & Strocco, F. (2017) Message safety in Dart. Sci. Comput. Program. 133, 51–73. Fähndrich, M., Aiken, M., Hawblitzel, C., Hodson, O., Hunt, G. C., Larus, J. R. & Levi, S. (2006)
Language support for fast and reliable message-based communication in Singularity OS. In
European Conference on Computer Systems (EuroSys). ACM, pp. 177–190. Fennell, L. & Thiemann, P. (2012). The blame theorem for a linear lambda calculus with type
dynamic. In Trends in Functional Programming, Loidl, H-W. & Peña, R. (eds). LNCS, vol. 7829. Springer, pp. 37–52. Fennell, L. & Thiemann, P. (2013) Gradual security typing with references. References In Principles of Programming Languages
(POPL). ACM, pp. 299–312. Girard, J.-Y. (1987) Linear logic. Theoret. Comput. Sci. 50(1), 1–101. ,
(
)
g
p
( ),
Gommerstadt, H., Jia, L. & Pfenning, F. (2018) Session-typed concurrent contracts. In Programming
Languages and Systems - 27th European Symposium on Programming, ESOP 2018, Held as
Part of the European Joint Conferences on Theory and Practice of Software, ETAPS 2018,
Thessaloniki, Greece, April 14–20, 2018, Proceedings, Ahmed, A. (ed). Lecture Notes in
Computer Science, vol. 10801. Springer, pp. 771–798. Greenberg, M., Pierce, B. C. & Weirich, S. (2010) Contracts made manifest. In Principles of
Programming Languages (POPL). ACM, pp. 353–364. Honda, K. (1993) Types for dyadic interaction. In International Conference on Concurrency Theory
(CONCUR). LNCS, vol. 715. Springer, pp. 509–523. Honda, K., Mukhamedov, A., Brown, G., Chen, T. & Yoshida, N. (2011) Scribbling interactions
with a formal foundation. In ICDCIT. LNCS, vol. 6536. Springer, pp. 55–75. Honda, K., Vasconcelos, V. & Kubo, M. (1998) Language primitives and type discipline for struc-
tured communication-based programming. In European Symposium on Programming (ESOP). LNCS. Springer, pp. 122–138. Honda, K., Yoshida, N. & Carbone, M. (2008) Multiparty asynchronous session types. In Principles
of Programming Languages (POPL). ACM, pp. 273–284. Honda, K., Yoshida, N. & Carbone, M. (2016) Multiparty asynchronous session types. J. ACM
63(1), 9. Hu, R., Kouzapas, D., Pernet, O., Yoshida, N. & Honda, K. (2010) Type-safe eventful sessions
in Java. In European Conference on Object-Oriented Programming (ECOOP). LNCS, vol. 6183. Springer, pp. 329–353. Hu, R. & Yoshida, N. (2016) Hybrid session verification through endpoint API generation. In Fundamental Approaches to Software Engineering (FASE). LNCS, vol. 9633. Springer,
pp. 401–418. Hu, R., Yoshida, N. & Honda, K. (2008) Session-based distributed programming in Java. In European Conference on Object-Oriented Programming (ECOOP). LNCS, vol. 5142. Springer,
pp. 516–541. Igarashi, A., Thiemann, P., Vasconcelos, V. T. & Wadler, P. (2017a) Gradual session types. PACMPL 1(ICFP), 38:1–38:28. Igarashi, Y., Sekiyama, T. & Igarashi, A. (2017b) On polymorphic gradual typing. PACMPL
1(ICFP), 40:1–40:29. Jafery, K. A. & Dunfield, J. (2017) Sums of uncertainty: Refinements go gradual. In Castagna,
G. & Gordon, A. D. (eds), Proceedings of the 44th ACM SIGPLAN Symposium on Principles of
Programming Languages, POPL 2017, Paris, France, January 18–20, 2017. ACM, pp. 804–817. Jespersen, T. B. L., Munksgaard, P. & Larsen, K. F. (2015) Session types for Rust. In Workshop on
Generic Programming (WGP). ACM, pp. 13–22. References In 2013 IEEE 26th
Computer Security Foundations Symposium, New Orleans, LA, USA, June 26–28, 2013. IEEE
Computer Society, pp. 224–239. Fennell, L. & Thiemann, P. (2016) LJGS: Gradual security types for object-oriented languages. In Krishnamurthi, S. & Lerner, B. S. (eds), 30th European Conference on Object-Oriented
Programming, ECOOP 2016, July 18–22, 2016, Rome, Italy. LIPIcs, vol. 56. Schloss Dagstuhl -
Leibniz-Zentrum fuer Informatik, pp. 9:1–9:26. Findler, R. B. & Felleisen, M. (2002) Contracts for higher-order functions. In International
Conference on Functional Programming (ICFP). ACM, pp. 48–59. lanagan, C. (2006) Hybrid type checking. In Principles of Programming Languages (POPL)
Morrisett, J. G. & Peyton Jones, S. L. (eds). ACM, pp. 245–256. Flanagan, C. (2006) Hybrid type checking. In Principles of Programming Languages (POPL),
Morrisett, J. G. & Peyton Jones, S. L. (eds). ACM, pp. 245–256. Fowler, S. (2016) An Erlang implementation of multiparty session actors. In Interaction and
Concurrency Experience, pp. 36–50. Morrisett, J. G. & Peyton Jones, S. L. (eds). ACM, pp. 245–256. Fowler, S. (2016) An Erlang implementation of multiparty session actors. In Interaction and
Concurrency Experience, pp. 36–50. Fowler, S. (2016) An Erlang implementation of multiparty session actors. In Interaction
Concurrency Experience, pp. 36–50. Franco, J. & Vasconcelos, V. T. (2013) A concurrent programming language with refined session
types. In SEFM. LNCS, vol. 8368. Springer, pp. 15–28. Garcia, R., Clark, A. M. & Tanter, É. (2016) Abstracting gradual typing. In Principles of
Programming Languages (POPL). ACM, pp. 429–442. É Garcia, R., Tanter, É., Wolff, R. & Aldrich, J. (2014) Foundations of typestate-oriented program-
ming. ACM Trans. Program. Lang. Syst. 36(4), 12:1–12:44. Gay, S. & Hole, M. (2005) Subtyping for session types in the pi calculus. Acta Informatica 42(2–3),
191–225. 48 A. Igarashi et al. Gay, S. & Vasconcelos, V. (2010) Linear type theory for asynchronous session types. J. Func
Program. 20(01), 19–50. Gay, S. & Vasconcelos, V. (2010) Linear type theory for asynchronous session types. J. Funct. Program. 20(01), 19–50. Gay, S. J. (2016) Subtyping supports safe session substitution. In A List of Successes That Can
Change the World - Essays Dedicated to Philip Wadler on the Occasion of His 60th Birthday,
Lindley, S., McBride, C., Trinder, P. W. & Sannella, D. (eds). Lecture Notes in Computer Science,
vol. 9600. Springer, pp. 95–108. Gay, S. J., Vasconcelos, V. T., Ravara, A., Gesbert, N. & Caldeira, A. Z. (2010) Modular ses-
sion types for distributed object-oriented programming. ms. https://doi.org/10.1017/S0956796819000169
idge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at References Jia, L., Gommerstadt, H. & Pfenning, F. (2016) Monitors and blame assignment for higher-
order session types. In Bodík, R. & Majumdar, R. (eds), Proceedings of the 43rd Annual
ACM SIGPLAN-SIGACT Symposium on Principles of Programming Languages, POPL 2016,
St. Petersburg, FL, USA, January 20–22, 2016. ACM, pp. 582–594. Kobayashi, N., Pierce, B. C. & Turner, D. N. (1999) Linearity and the pi-calculus. ACM Trans. Program. Lang. Syst. 21(5), 914–947. 49 Gradual session types Kouzapas, D., Dardha, O., Perera, R. & Gay, S. J. (2016) Typechecking protocols with Mungo and
StMungo. In Principles and Practice of Declarative Programming (PPDP). ACM, pp. 146–159. É Lehmann, N. & Tanter, É. (2017) Gradual refinement types. In Proceedings of the 44th ACM
SIGPLAN Symposium on Principles of Programming Languages, POPL 2017, Paris, France,
January 18–20, 2017, Castagna, G. & Gordon, A. D. (eds). ACM, pp. 775–788. Lindley, S. & Morris, J. G. (2016a) Embedding session types in Haskell. In Symposium on Haskell. ACM, pp. 133–145. Lindley, S. & Morris, J. G. (2016b) Talking bananas: Structural recursion for session types. In International Conference on Functional Programming (ICFP). ACM, pp. 434–447. Lindley, S. & Morris, J. G. (2017) Lightweight functional session types. In Behavioural Types: Fr
Theory to Tools. River Publishers. Melgratti, H. C. & Padovani, L. (2017) Chaperone contracts for higher-order sessions. PACMPL
1(ICFP), 35:1–35:29. Milner, R., Parrow, J. & Walker, D. (1992) A calculus of mobile processes, I. Inform. Comput. 100(1), 1–40. Mostrous, D. & Vasconcelos, V. T. (2011) Session typing for a featherweight Erlang. In Coordination Models and Languages (COORDINATION). LNCS, vol. 6721. Springer,
pp. 95–109. Neykova, R. & Yoshida, N. (2014) Multiparty session actors. In Coordination Models and Languages
(COORDINATION). LNCS, vol. 8459. Springer, pp. 131–146. Neykova, R. & Yoshida, N. (2017) Let it recover: Multiparty protocol-induced recovery. In International Conference on Compiler Construction (CC). ACM, pp. 98–108. Neykova, R., Yoshida, N. & Hu, R. (2013) SPY: Local verification of global protocols. In International Conference on Runtime Verification (RV). LNCS, vol. 8174. Springer,
pp. 358–363. Ng, N., Yoshida, N. & Honda, K. (2012) Multiparty Session C: Safe parallel programming with
message optimisation. In International Conference on Modelling Techniques and Tools for
Computer Performance Evaluation (TOOLS). LNCS, vol. 7304. Springer, pp. 202–218. Norell, U. (2009) Dependently typed programming in Agda. In Proceedings of the 4th International
Workshop on Types in Language Design and Implementation. TLDI’09. ACM, pp. 1–2. Orchard, D. & Yoshida, N. References (eds), A List of Successes That Can Change the
World - Essays Dedicated to Philip Wadler on the Occasion of His 60th Birthday. Lecture Notes
in Computer Science, vol. 9600. Springer, pp. 388–410. Siek, J. G., Vitousek, M. M., Cimini, M. & Boyland, J. T. (2015b) Refined criteria for gradual typing. In Summit on Advances in Programming Languages (SNAPL). LIPIcs, vol. 32. Schloss Dagstuhl,
pp. 274–293. The Coq Development Team (2019) The coq proof assistant, version 8.9.0. The Dart Team (2014) Dart programming language specification. Google, 1.2 edition. Thiemann, P. (2014) Session types with gradual typing. In TGC. LNCS, vol. 8902. Springer,
pp. 144–158. Thiemann, P. & Fennell, L. (2014) Gradual typing for annotated type systems. In Programming
Languages and Systems - 23rd European Symposium on Programming, ESOP 2014, Held as
Part of the European Joint Conferences on Theory and Practice of Software, ETAPS 2014,
Grenoble, France, April 5–13, 2014, Proceedings, Shao, Z. (ed). Lecture Notes in Computer
Science, vol. 8410. Springer, pp. 47–66. Tobin-Hochstadt, S. & Felleisen, M. (2008) The design and implementation of typed Scheme. In
Proceedings of the 35th ACM SIGPLAN-SIGACT Symposium on Principles of Programming
Languages, POPL 2008, San Francisco, California, USA, January 7–12, 2008, Necula, G. C. &
Wadler, P. (eds). ACM, pp. 395–406. (
)
pp
Toninho, B., Caires, L. & Pfenning, F. (2013) Higher-order processes, functions, and sessions:
A monadic integration. In European Symposium on Programming (ESOP). LNCS, vol. 7792. Springer, pp. 350–369. p
g
pp
Toninho, B. & Yoshida, N. (2018) Depending on session-typed processes. In FoSSaCS. Lecture
Notes in Computer Science, vol. 10803. Springer, pp. 128–145. Toro, M., Garcia, R. & Tanter, É. (2018) Type-driven gradual security with references. ACM Trans. Program. Lang. Syst. 40(4), 16:1–16:55. g
g
y
( )
Toro, M., Labrada, E. & Tanter, É. (2019) Gradual parametricity, revisited. PACMPL 3(POPL),
17:1–17:30. Tov, J. A. & Pucella, R. (2010) Stateful contracts for affine types. In European Symposium on
Programming (ESOP). LNCS, vol. 6012. Springer, pp. 550–569. Vasconcelos, V. T. (2012) Fundamentals of session types. Inform. Comput. 217, 52–70. É Vazou, N., Tanter, É. & Horn, D. V. (2018) Gradual liquid type inference. PACMPL 2(OOPSLA),
132:1–132:25. Verlaguet, J. (2013) Facebook: Analysing PHP statically. In Workshop on Commercial Uses o
Functional Programming (CUFP). Vitousek, M. M., Swords, C. & Siek, J. G. (2017) Big types in little runtime: Open-world soundness
and collaborative blame for gradual type systems. References (2016) Effects as sessions, sessions as effects. In Principles of
Programming Languages (POPL). ACM, pp. 568–581. g
g
g
g
(
)
pp
Ou, X., Tan, G., Mandelbaum, Y. & Walker, D. (2004) Dynamic typing with dependent types. In
IFIP International Conference on Theoretical Computer Science, vol. 155. Springer, pp. 437–450. P d
i L (2017) A i
l lib
i
l
t ti
f bi
i
J F
t P
27
4 p
p
g
pp
Padovani, L. (2017) A simple library implementation of binary sessions. J. Funct. Program 27, e4 Pfenning, F. & Griffith, D. (2015) Polarized substructural session types. In International Conference
on Foundations of Software Science and Computation Structures. LNCS, vol. 9034. Springer,
pp. 3–22. Pierce, B. C. (2002) Types and Programming Languages. MIT Press. Polakow, J. (2015) Embedding a full linear lambda calculus in Haskell. In Symposium on Haskell. ACM, pp. 177–188. Pucella, R. & Tov, J. A. (2008) Haskell session types with (almost) no class. In Symposium
Haskell. ACM, pp. 25–36. Sackman, M. & Eisenbach, S. (2008) Session Types in Haskell: Updating Message Passing for
21st Century. Available at: https://spiral.imperial.ac.uk:8443/handle/10044/1/5918. Scalas, A. & Yoshida, N. (2016) Lightweight session programming in Scala. In European Conference
on Object-Oriented Programming (ECOOP). LIPIcs. Schloss Dagstuhl, pp. 21:1–21:28. É Schwerter, F. B., Garcia, R. & Tanter, É. (2016) Gradual type-and-effect systems. J. Funct. Program. 26, e19. Sergey, I. & Clarke, D. (2012) Gradual ownership types. In Programming Languages and Systems -
21st European Symposium on Programming, ESOP 2012, Held as Part of the European Joint
Conferences on Theory and Practice of Software, ETAPS 2012, Tallinn, Estonia, March 24–April
1, 2012. Proceedings, Seidl, H. (ed). Lecture Notes in Computer Science, vol. 7211. Springer,
pp. 579–599. 50 A. Igarashi et al. Siek, J., Thiemann, P. & Wadler, P. (2015a) Blame and coercion: Together again for the first time. In Programming Language Design and Implementation (PLDI), ACM, pp. 425–435. Siek, J. G., & Taha, W. (2006) Gradual typing for functional languages. In Scheme and Functional
Programming Workshop (Scheme), pp. 81–92. Available at: http://scheme2006.cs.uchicago.edu
and the paper URL is http://scheme2006.cs.uchicago.edu/13-siek.pdf Siek, J. G. & Taha, W. (2007) Gradual typing for objects. In ECOOP. Lecture Notes in Computer
Science, vol. 4609. Springer, pp. 2–27. Siek, J. G. & Tobin-Hochstadt, S. (2016) The recursive union of some gradual types. In Lindley,
S., McBride, C., Trinder, P. W. & Sannella, D. References Wolff, R., Garcia, R., Tanter, É. & Aldrich, J. (2011) Gradual typestate. In European Conference on
Object-Oriented Programming (ECOOP). LNCS, vol. 6813. Springer, pp. 459–483. Wolff, R., Garcia, R., Tanter, É. & Aldrich, J. (2011) Gradual typestate. In European Conference on
Object-Oriented Programming (ECOOP). LNCS, vol. 6813. Springer, pp. 459–483. Yoshida, N., Hu, R., Neykova, R. & Ng, N. (2014) The Scribble protocol language. In International
Symposium on Trustworthy Global Computing LNCS vol 8358 Springer pp 22–41 Wolff, R., Garcia, R., Tanter, É. & Aldrich, J. (2011) Gradual typestate. In European Conference on
Object-Oriented Programming (ECOOP). LNCS, vol. 6813. Springer, pp. 459–483. Yoshida, N., Hu, R., Neykova, R. & Ng, N. (2014) The Scribble protocol language. In International
Symposium on Trustworthy Global Computing. LNCS, vol. 8358. Springer, pp. 22–41. Yoshida, N. & Vasconcelos, V. (2007) Language primitives and type discipline for structured
communication-based programming revisited: Two systems for higher-order session communi- oshida, N., Hu, R., Neykova, R. & Ng, N. (2014) The Scribble protocol language. In International
Symposium on Trustworthy Global Computing. LNCS, vol. 8358. Springer, pp. 22–41. Yoshida, N. & Vasconcelos, V. (2007) Language primitives and type discipline for structured
communication-based programming revisited: Two systems for higher-order session communi-
cation. Entcs 171(4), 73–93. Yoshida, N. & Vasconcelos, V. (2007) Language primitives and type discipline for structured
communication-based programming revisited: Two systems for higher-order session communi-
cation. Entcs 171(4), 73–93. References In Proceedings of the 44th ACM SIGPLAN
Symposium on Principles of Programming Languages, POPL 2017, Paris, France, January 18–20,
2017, Castagna, G. & Gordon, A. D. (eds). ACM, pp. 762–774. Wadler, P. (2012) Propositions as sessions. In International Conference on Functional Programming
(ICFP). ACM, pp. 273–286. Wadler, P. (2014) Propositions as sessions. J. Funct. Program. 24(2–3), 384–418. Wadler, P. (2015) A complement to blame. In 1st Summit on Advances in Programming Languages
(SNAPL). LIPIcs, vol. 32. Schloss Dagstuhl, pp. 309–320. Wadler, P. & Findler, R. B. (2009) Well-typed programs can’t be blamed. In European Symposium
on Programming (ESOP). LNCS, vol. 5502. Springer, pp. 1–16. 51 Gradual session types Walker, D. (2005) Substructural type systems. In Advanced Topics in Types and Programming
Languages. MIT Press, pp. 3–43. Walker, D. (2005) Substructural type systems. In Advanced Topics in Types and Programming
Languages. MIT Press, pp. 3–43. Walker, D. (2005) Substructural type systems. In Advanced Topics in Types and Programming
Languages. MIT Press, pp. 3–43. Williams, J., Morris, J. G., Wadler, P. & Zalewski, J. (2017) Mixed messages: Measuring con-
formance and non-interference in TypeScript. In European Conference on Object-Oriented
Programming (ECOOP). LIPIcs, vol. 74. Dagstuhl, Germany: Schloss Dagstuhl, pp. 28:1–28:29. Willsey M
Prabhu R & Pfenning F (2017) Design and implementation of concurrent C0 In Williams, J., Morris, J. G., Wadler, P. & Zalewski, J. (2017) Mixed messages: Measuring con-
formance and non-interference in TypeScript. In European Conference on Object-Oriented
Programming (ECOOP). LIPIcs, vol. 74. Dagstuhl, Germany: Schloss Dagstuhl, pp. 28:1–28:29. Willsey, M., Prabhu, R. & Pfenning, F. (2017) Design and implementation of concurrent C0. In
International Workshop on Linearity. EPTCS, vol. 238, pp. 73–82. Williams, J., Morris, J. G., Wadler, P. & Zalewski, J. (2017) Mixed messages: Measuring con
formance and non-interference in TypeScript. In European Conference on Object-Oriented
Programming (ECOOP). LIPIcs, vol. 74. Dagstuhl, Germany: Schloss Dagstuhl, pp. 28:1–28:29. Willsey, M., Prabhu, R. & Pfenning, F. (2017) Design and implementation of concurrent C0. In
International Workshop on Linearity. EPTCS, vol. 238, pp. 73–82. Wolff, R., Garcia, R., Tanter, É. & Aldrich, J. (2011) Gradual typestate. In European Conference on
Obj
t O i
t d P
i
(ECOOP) LNCS
l 6813 S
i
459 483 Programming (ECOOP). LIPIcs, vol. 74. Dagstuhl, Germany: Schloss Dagstuhl, pp. 28:1–28:29. Willsey, M., Prabhu, R. & Pfenning, F. (2017) Design and implementation of concurrent C0. In
International Workshop on Linearity. EPTCS, vol. 238, pp. 73–82. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at Appendix: Type-checking algorithm for the external language We give a type-checking algorithm for GGVe and show that it is correct. The type-checking
algorithm is slightly involved due to linearity: CHECKEXPR(, e) outputs a pair of type T
and a set X of variables, containing the linear variables occurring free in e. function CHECKEXPR(, e) function CHECKEXPR(, e)
case e of function CHECKEXPR(, e)
case e of | () ⇒return unit, ∅ | e1 e2 ⇒ https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Camb 52 A. Igarashi et al. g
| (e1, e2)m ⇒
T1, X := CHECKEXPR(, e1); T2, Y := CHECKEXPR(, e2)
assert X ∩Y = ∅
if m = un then assert un(T1) and un(T2)
return T1 ×m T2, X ∪Y
| let x1, x2 = e1 in e2 ⇒
T, Y := CHECKEXPR(, e1)
T1 ×m T2 := MATCHINGPROD(T)
U, Z := CHECKEXPR((, x : T1, y : T2), e2)
if lin(T1) then
assert x1 ∈Z
Z := Z \ {x1}
if lin(T2) then
assert x2 ∈Z
Z := Z \ {x2}
assert Y ∩Z = ∅
return U, Y ∪Z
| fork e1 ⇒
T, X := CHECKEXPR(, e1)
assert T ∼unit
return unit, X
| new S ⇒return S ×lin S, ∅
| send e1 e2 ⇒
T1, X := CHECKEXPR(, e1); T2, Y := CHECKEXPR(, e2)
assert X ∩Y = ∅
!T3. S := MATCHINGSEND(T2)
assert T1 ≲T3
return S, X ∪Y
| receive e1 ⇒
T1, X := CHECKEXPR(, e1)
?T2. S := MATCHINGRECEIVE(T1)
return T2 ×lin S, X
| select lj e1 ⇒
T, X := CHECKEXPR(, e1)
⊕{lj : Sj} := MATCHINGSELECT(T, lj)
return Sj, X
| case e0 of {l1 : x1. e1, . . . , lk : xk. ek} ⇒
T, X := CHECKEXPR(, e0)
&{l1 : R1, . . . , lk : Rk} := MATCHINGCASE(T, {l1, . . . https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Camb Appendix: Type-checking algorithm for the external language return unit, X
| wait e1 ⇒
T, X := CHECKEXPR(, e1)
assert T ∼end? return unit, X function MATCHINGFUN(T)
case T of
| T1 →m T2 ⇒return T1 →m T2
| ⋆⇒return ⋆→lin ⋆
| _ ⇒error
function MATCHINGPROD(T)
case T of
| T1 ×m T2 ⇒return T1 ×m T2
| ⋆⇒return ⋆×lin ⋆
| _ ⇒error
function MATCHINGSEND(T)
case T of
| !T′. S ⇒return !T′. S
| ⋆⃝| ⋆⇒return !⋆. ⋆⃝
| _ ⇒error
function MATCHINGRECEIVE(T)
case T of
| !T′. S ⇒return !T′. S
| ⋆⃝| ⋆⇒return !⋆. ⋆⃝
| _ ⇒error
function MATCHINGSELECT(T, lj)
case T of
| ⊕{li : Si}i∈I ⇒
assert j ∈I
return ⊕{lj : Sj}
| ⋆⃝| ⋆⇒return ⊕{lj : ⋆⃝}
| _ ⇒error
function MATCHINGCASE(T, {lj}j∈J)
case T of Appendix: Type-checking algorithm for the external language , lk})
for j = 1 to k do
Uj, Yj := CHECKEXPR((, xj : Rj), ej)
assert xj ∈Yj | (e1, e2)m ⇒
T1, X := CHECKEXPR(, e1); T2, Y := CHECKEXPR(, e2)
assert X ∩Y = ∅
if m = un then assert un(T1) and un(T2)
return T1 ×m T2, X ∪Y
| let x1, x2 = e1 in e2 ⇒
T, Y := CHECKEXPR(, e1)
T1 ×m T2 := MATCHINGPROD(T)
U, Z := CHECKEXPR((, x : T1, y : T2), e2)
if lin(T1) then
assert x1 ∈Z
Z := Z \ {x1}
if lin(T2) then
assert x2 ∈Z
Z := Z \ {x2}
assert Y ∩Z = ∅
return U, Y ∪Z
| fork e1 ⇒
T, X := CHECKEXPR(, e1)
assert T ∼unit
return unit, X
| new S ⇒return S ×lin S, ∅
| send e1 e2 ⇒
T1, X := CHECKEXPR(, e1); T2, Y := CHECKEXPR(, e2)
assert X ∩Y = ∅
!T3. S := MATCHINGSEND(T2)
assert T1 ≲T3
return S, X ∪Y
| receive e1 ⇒
T1, X := CHECKEXPR(, e1)
?T2. S := MATCHINGRECEIVE(T1)
return T2 ×lin S, X
| select lj e1 ⇒
T, X := CHECKEXPR(, e1)
⊕{lj : Sj} := MATCHINGSELECT(T, lj)
return Sj, X
| case e0 of {l1 : x1. e1, . . . , lk : xk. ek} ⇒
T, X := CHECKEXPR(, e0)
&{l1 : R1, . . . , lk : Rk} := MATCHINGCASE(T, {l1, . . . , lk})
for j = 1 to k do
Uj, Yj := CHECKEXPR((, xj : Rj), ej)
assert x ∈Y | fork e1 ⇒ | new S ⇒return S ×lin S, ∅ | receive e1 ⇒ T1, X := CHECKEXPR(, e1) ?T2. S := MATCHINGRECEIVE(T1) | case e0 of {l1 : x1. e1, . . . , lk : xk. ek} ⇒ T, X := CHECKEXPR(, e0) &{l1 : R1, . . . , lk : Rk} := MATCHINGCASE(T, {l1, . . . , lk}) for j = 1 to k do
Uj, Yj := CHECKEXPR((, xj : Rj), ej)
assert xj ∈Yj
Yj := Yj \ {xj} 53 Gradual session types assert Y1 = · · · = Yk(=: Y)
U := U1 ∨. . . ∨Uk
assert X ∩Y = ∅
return U, X ∪Y
| close e1 ⇒
T, X := CHECKEXPR(, e1)
assert T ∼end! function MATCHINGFUN(T)
case T of | T1 →m T2 ⇒return T1 →m T2 A. Igarashi et al. 54 | &{li : Si}i∈I ⇒
if I ⊆J then return &{li : Si}i∈I ∪{lj : ⋆⃝}j∈J\I
else error | &{li : Si}i∈I ⇒
if I ⊆J then return &{li : Si}i∈I ∪{lj : ⋆⃝}j∈J\I
else error | ⋆⃝| ⋆⇒return &{lj : ⋆⃝}j∈J
| _ ⇒error | ⋆⃝| ⋆⇒return &{lj : ⋆⃝}j∈J | ⋆⃝| ⋆⇒return &{lj : ⋆⃝}j∈J | ⋆⃝| ⋆⇒return &{lj : ⋆⃝}j∈J | _ ⇒error Theorem 11 states soundness of the type-checking algorithm. A few lemmas are required
in preparation. Let rm(, X) denote the operation that removes variables X from a type
environment . Lemma 19. Suppose y: U ∈ with lin(U). Lemma 19. Suppose y: U ∈ with lin(U). If CHECKEXPR(, e) = T, X and y /∈X, then CHECKEXPR(rm(, {y}), e) = T, X. Lemma 19. Suppose y: U ∈ with lin(U). If CHECKEXPR(, e) = T, X and y /∈X, then CHECKEXPR(rm(, {y}), e) = T, X. Proof. By induction on e, we show one important case as follows. Case e = e1 e2: We are given Case e = e1 e2: We are given (y) = U
lin(U)
CHECKEXPR(, e1 e2) = T, X
y /∈X
By the definition of the algorithm, CHECKEXPR(, ei) = Ti, Xi for i = 1, 2 and
T1 ▷T11 →m T12
T2 ≲T11
T = T12
X = X1 ⊎X2 (y) = U
lin(U)
CHECKEXPR(, e1 e2) = T, X
y /∈X
ion of the algorithm, CHECKEXPR(, ei) = Ti, Xi for i = 1, 2 and
T1 ▷T11 →m T12
T2 ≲T11
T = T12
X = X1 ⊎X2 T1 ▷T11 →m T12
T2 ≲T11
T = T12
X = X1 ⊎X2 Since y /∈X, we have y /∈X1 and y /∈X2. By CHECKEXPR(, ei) = Ti, Xi and y /∈Xi and the
IH for i = 1, 2, we have Since y /∈X, we have y /∈X1 and y /∈X2. By CHECKEXPR(, ei) = Ti, Xi and y /∈Xi and the
IH for i = 1, 2, we have CHECKEXPR(rm(, {y}), e1) = T1, X1
CHECKEXPR(rm(, {y}), e2) = T2, X2
Thus, by the definition of the algorithm, CHECKEXPR(rm(, {y}), e1 e2) = T, X. □ Lemma 20. Suppose flv() = X1 ⊎X2. CHECKEXPR((, x : T1), e1) = T2, {x}
X = ∅ So, flv() = X = ∅. By lin(T1), we have flv(, x : T1) = {x}. Thus, by the IH, we have , x So, flv() = X = ∅. By lin(T1), we have flv(, x : T1) = {x}. Thus, by the IH, we have , x :
T1 ⊢e1 : T2. Here, by flv() = ∅, we have un(). So, un:>(). We finish by So, flv() = X = ∅. By lin(T1), we have flv(, x : T1) = {x}. Thus, by the IH, we h
T1 ⊢e1 : T2. Here, by flv() = ∅, we have un(). So, un:>(). We finish by T1 ⊢e1 : T2. Here, by flv() = ∅, we have un(). So, un:>(). We finish by , x : T1 ⊢e1 : T2
un:>() and the abstraction rule. function MATCHINGFUN(T)
case T of If 1 = rm(, X2) and 2 = rm(, X1), then =
1 ◦2 and flv(1) = X1 and flv(2) = X2. Proof. By induction on . □ □ Proof. By induction on . Theorem 11 (Soundness of the type-checking algorithm). If CHECKEXPR(, e) = T, X
and flv() = X, then ⊢e e : T. Theorem 11 (Soundness of the type-checking algorithm). If CHECKEXPR(, e) = T, X
and flv() = X, then ⊢e e : T. Proof. By induction on e, we show main cases as follows. Proof. By induction on e, we show main cases as follows. Proof. By induction on e, we show main cases as follows. Case e = λmx:T1. e1: We are given Case e = λmx:T1. e1: We are given CHECKEXPR(, λmx:T1. e1) = T, X
flv() = X CHECKEXPR(, λmx:T1. e1) = T, X
flv() = X We consider only when m = un and lin(T1). By the definition of the algorithm, We consider only when m = un and lin(T1). By the definition of the algorithm CHECKEXPR((, x : T1), e1) = T2, {x}
X = ∅ and the abstraction rule. and the abstraction rule. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at 55 Gradual session types Case e = e1 e2: We are given Case e = e1 e2: We are given CHECKEXPR(, e1 e2) = T, X
flv() = X
By the definition of the algorithm, CHECKEXPR(, ei) = Ti, Xi for i = 1, 2 and
T1 ▷T11 →m T12
T2 ≲T11
T = T12
X = X1 ⊎X2 Let 1 = rm(, X2) and 2 = rm(, X1). By Lemma 20, = 1 ◦2 and flv(1) =
X1
and flv(2) = X2. By CHECKEXPR(, ei) = Ti, Xi
and Lemma 19, we have
CHECKEXPR(i, ei) = Ti, Xi for i = 1, 2. By flv(i) = Xi and CHECKEXPR(i, ei) = Ti, Xi
and the IH, we have i ⊢ei : Ti for i = 1, 2. We finish by Let 1 = rm(, X2) and 2 = rm(, X1). By Lemma 20, = 1 ◦2 and flv(1) =
X1
and flv(2) = X2. By CHECKEXPR(, ei) = Ti, Xi
and Lemma 19, we have
CHECKEXPR(i, ei) = Ti, Xi for i = 1, 2. By flv(i) = Xi and CHECKEXPR(i, ei) = Ti, Xi
and the IH, we have i ⊢ei : Ti for i = 1, 2. We finish by 1 ⊢e1 : T1
2 ⊢e2 : T2
T1 ▷T11 →m T12
T2 ≲T11
he application rule. 1 ⊢e1 : T1
2 ⊢e2 : T2
T1 ▷T11 →m T12
T2 ≲T11 □ and the application rule. and the application rule. We will also show the converse of the theorem above. Completeness states that
CHECKEXPR(, e) computes a minimal type with respect to negative subtyping. Theorem 12 (Completeness of the type-checking algorithm). If ⊢e e : T, then
CHECKEXPR(, e) = T′, X and T′ <:−T and flv() = X for some T′. Theorem 12 (Completeness of the type-checking algorithm). If ⊢e e : T, then
CHECKEXPR(, e) = T′, X and T′ <:−T and flv() = X for some T′. To prove this theorem, we need a stronger statement, namely Lemma 24. We define
environment positive consistent subtyping, written ′ <:−, as dom() ⊆dom(′) and
(x) <:−′(x), for any x ∈dom(). Then, the theorem follows from the fact that <:−on
type environments is reflexive. and the abstraction rule. We start with a few lemmas about <:−. Lemma 21. 1. If T′
1 <:−T1 and T1 ▷T11 →m T12, then there exist some T′
11, T′
12, and n such that
MATCHINGFUN(T′
1) = T′
11 →n T′
12 and T′
11 →n T′
12 <:−T11 →m T12. 2. If T′
1 <:−T1 and T1 ▷T11 ×m T12, then there exist some T′
11, T′
12, and n such that
MATCHINGPROD(T′
1) = T′
11 ×n T′
12 and T′
11 ×n T′
12 <:−T11 ×m T12. 3. If T′
1 <:−T1 and T1 ▷!T11. S12, then there exist some T′
11 and S′
12 such that
MATCHINGSEND(T′
1) = !T′
11. S′
12 and !T′
11. S′
12 <:−!T11. S12. 4. If T′
1 <:−T1 and T1 ▷?T11. S12, then there exist some T′
11 and S′
12 such that
MATCHINGRECEIVE(T′
1) = ?T′
11. S′
12 and ?T′
11. S′
12 <:−?T11. S12. 5. If T′
1 <:−T1 and T1 ▷⊕{lj : Sj}, then there exist some S′
j such that
MATCHINGSELECT(T′
1, lj) = ⊕{lj : S′
j} and ⊕{lj : S′
j} <:−⊕{lj : Sj}. j
j
6. If T′
1 <:−T1 and T1 ▷&{l1 : S1, . . . , lk : Sk}, then there exist some S′
1, . . . , S′
k
such that MATCHINGCASE(T′
1, {l1, . . . , lk}) = &{l1 : S′
1, . . . , lk : S′
k} and &{l1 :
S′
1, . . . , lk : S′
k} <:−&{l1 : S1, . . . , lk : Sk}. □ Proof. By induction on e, we show main cases as follows. Proof. By induction on e, we show main cases as follows. Proof. By induction on e, we show main cases as follows. Case e = e1 e2: By inversion of the typing relation, 1 ⊢e e1 : T1 and 2 ⊢e e2 : T2 and
= 1 ◦2 and T1 ▷T11 →m T12 and T2 ≲T11 for some 1, 2, T1, T2, T11, T12, and m. It is easy to show ′ <:−1 and ′ <:−2. By the induction hypothesis, for some T′
1,
T′
2, X, and Y, T′
1, X = CHECKEXPR(′, e1) and T′
1 <:−T1 and flv(1) = X and T′
2, Y =
CHECKEXPR(′, e2) and T′
2 <:−T2 and flv(2) = Y. Since 1 ◦2 is well defined, X ∩Y
must be ∅. By Lemma 21, T′
11 →n T′
12 = MATCHINGFUN(T′
1) and T′
11 →n T′
12 <:−T11 →m
T12 for some T′
11 and T′
12. By inversion of <:−, we have T11 <:+ T′
11 and T′
12 <:−T12. Then,
T′
2 ≲T′
11 is shown by Lemma 23. It is easy to show X ∪Y = flv() because = 1 ◦2. Finally, T′
12 <:−T12 finishes the case. Case e = case e0 of {lj : xj. ej}j∈J: By inversion of the typing relation, we have 1 ⊢e e0 :
T0 and T0 ▷&{lj : Rj}j∈J and (2, xj : Rj ⊢e ej : Uj)j∈J and T = ∨{Uj}j∈J and = 1 ◦2
for some 1, 2, Rj, and Uj (for j ∈J). It is easy to show ′ <:−1 and ′ <:−2. By
the induction hypothesis, T′
0, X = CHECKEXPR(′, e0) and T′
0 <:−T0 and flv(1) = X for
some T′
0 and X. By Lemma 21, MATCHINGCASE(T′
0, {l1, . . . , lk}) = &{l1 : R′
1, . . . , lk : R′
k}
and &{lj : R′
j}j∈J <:−&{lj : Rj}j∈J for some (R′
j)j∈J. By inversion of <:−, we have (R′
j <:−
Rj)j∈J. By the induction hypothesis, for any j ∈J, there exist U′
j and Yj such that U′
j, Yj =
CHECKEXPR((′, xj : R′
j), ej) and U′
j <:−Uj and flv(2, xj : Rj) = Yj. Proof. By case analysis on T ▷U. Proof. By case analysis on T ▷U. Lemma 22. If T <:−U and un(U), then un(T). Proof. By case analysis on T <:−U. □ A. Igarashi et al. 56 Lemma 23. 1. If T1 <:−T2 and T2 ≲T3, then T1 ≲T3. 2. If T1 ≲T2 and T2 <:+ T3, then T1 ≲T3. 1. If T1 <:−T2 and T2 ≲T3, then T1 ≲T3. 1. If T1 <:−T2 and T2 ≲T3, then T1 ≲T3. 1. If T1 <:−T2 and T2 ≲T3, then T1 ≲T3. 2. If T1 ≲T2 and T2 <:+ T3, then T1 ≲T3. 1. If T1 <: T2 and T2 ≲T3, then T1 ≲T3. 2. If T1 ≲T2 and T2 <:+ T3, then T1 ≲T3. 2. If T1 ≲T2 and T2 <:+ T3, then T1 ≲T3. Proof. Both items are proved by simultaneous induction on ≲. □ Proof. Both items are proved by simultaneous induction on ≲. Proof. Both items are proved by simultaneous induction on ≲. Lemma 24. If ⊢e e : T and ′ <:−, then there exists T′ such that CHECKEXPR(′, e) =
T′, X and T′ <:−T and flv() = X. Lemma 24. If ⊢e e : T and ′ <:−, then there exists T′ such that CHECKEXPR(′, e) =
T′, X and T′ <:−T and flv() = X. https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0956796819000169
Downloaded from https://www.cambridge.org/core. University of Edinburgh, on 25 Nov 2019 at 10:25:18, subject to the Camb Proof. By induction on e, we show main cases as follows. It is easy to show that
xj ∈Yj for any j ∈J and Y1 = · · · = Yk and X ∩Y1 = ∅because Rj is linear and 1 ◦2 is
well defined. It is also easy to show X ∪(Y1 \ {x1}) = flv() because = 1 ◦2. Finally,
∨{U′
j}j∈J <:−∨{Uj}j∈J is shown by Lemma 15. □
|
https://openalex.org/W1772411330
|
https://europepmc.org/articles/pmc4698573?pdf=render
|
English
| null |
Chromosomal distribution of microsatellite repeats in Amazon cichlids genome (Pisces, Cichlidae)
|
Comparative cytogenetics
| 2,015
|
cc-by
| 4,967
|
Chromosomal distribution of microsatellite repeats in
Amazon cichlids genome (Pisces, Cichlidae) Carlos Henrique Schneider1, Maria Claudia Gross1, Maria Leandra Terencio2,
Édika Sabrina Girão Mitozo de Tavares1, Cesar Martins3, Eliana Feldberg4 Carlos Henrique Schneider1, Maria Claudia Gross1, Maria Leandra Terencio2,
Édika Sabrina Girão Mitozo de Tavares1, Cesar Martins3, Eliana Feldberg4 1 Universidade Federal do Amazonas, Instituto de Ciências Biológicas, Departamento de Genética, Laboratório
de Citogenômica Animal, Av. General Rodrigo Otávio, 3000, Japiim, Zip code 69077-000 Manaus, AM, Bra
zil 2 Universidade Federal da Integração Latino Americana, Laboratório de Genética, Av. Tarquínio Joslin dos
Santos, 1000, Jardim Universitário, Zip code 85857-190, Foz do Iguaçu, PR, Brazil 3 Universidade Estadual
Paulista Júlio de Mesquita Filho – UNESP, Instituto de Biociências, Departamento de Morfologia, Laboratório
Genômica Integrativa, Rubião Junior, Zip code 18618-000 Botucatu, SP, Brazil 4 Instituto Nacional de
Pesquisas da Amazônia, Laboratório de Genética Animal, Av. André Araújo, 2936 Zip Code 69077-000,
Manaus, AM, Brazil orresponding author: Carlos Henrique Schneider (schneider.carloshenrique@gmail.com) Corresponding author: Carlos Henrique Schneider (schneider.carloshenrique@gmail.com)
Academic editor: E. Gornung | Received 3 July 2015 | Accepted 21 August 2014 | Published 14 September 2015
http://zoobank.org/663D53CE-DFC1-47D1-B0D7-782AB3168A03
Citation: Schneider CH, Gross MC, Leandra Terencio M, de Tavares ÉSGM, Martins C, Feldberg E (2015) Chromosomal
distribution of microsatellite repeats in Amazon cichlids genome (Pisces, Cichlidae). Comparative Cytogenetics 9(4):
595–605. doi: 10.3897/CompCytogen.v9i4.5582 Corresponding author: Carlos Henrique Schneider (schneider.carloshenrique@gmail.com)
Academic editor: E. Gornung | Received 3 July 2015 | Accepted 21 August 2014 | Published 14 September 2015
http://zoobank.org/663D53CE-DFC1-47D1-B0D7-782AB3168A03
Citation: Schneider CH, Gross MC, Leandra Terencio M, de Tavares ÉSGM, Martins C, Feldberg E (2015) Chromosomal
distribution of microsatellite repeats in Amazon cichlids genome (Pisces, Cichlidae). Comparative Cytogenetics 9(4):
595–605. doi: 10.3897/CompCytogen.v9i4.5582 Corresponding author: Carlos Henrique Schneider (schneider.carloshenrique@gmail.com)
Academic editor: E. Gornung | Received 3 July 2015 | Accepted 21 August 2014 | Published 14 September 2015
http://zoobank.org/663D53CE-DFC1-47D1-B0D7-782AB3168A03
Citation: Schneider CH, Gross MC, Leandra Terencio M, de Tavares ÉSGM, Martins C, Feldberg E (2015) Chromosomal
distribution of microsatellite repeats in Amazon cichlids genome (Pisces, Cichlidae). Comparative Cytogenetics 9(4):
595–605. doi: 10.3897/CompCytogen.v9i4.5582 Citation: Schneider CH, Gross MC, Leandra Terencio M, de Tavares ÉSGM, Martins C, Feldberg E (2015) Chromosomal
distribution of microsatellite repeats in Amazon cichlids genome (Pisces, Cichlidae). Comparative Cytogenetics 9(4):
595–605. doi: 10.3897/CompCytogen.v9i4.5582 Copyright Carlos Henrique Schneider et al. This is an open access article distributed under the terms of the Creative Commons Attribution License
(CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Chromosomal distri
CompCytogen 9(4): 595–605 (2015)
doi: 10.3897/CompCytogen.v9i4.5582
http://compcytogen.pensoft.net Chromosomal distri
CompCytogen 9(4): 595–605 (2015)
doi: 10.3897/CompCytogen.v9i4.5582
http://compcytogen.pensoft.net microsatellite repeats
RESEARCH ARTICLE Introduction The fish family Cichlidae exhibits high species richness with approximately 3,000 spe
cies distributed in Central and South America, Africa, and South India (Kullander
1998, Kocher 2004). The evolution of this family is characterized by repeated adaptive
radiation and sympatric speciation (Schliewen et al. 1994, Seehausen 2006). Moreo
ver, these fish are considered to be an excellent model for evolutionary studies because
of their morphological and behavioral adaptations to a wide diversity of explored eco
logical niches (Lowe-McConnell 1991). Approximately 400 species have been identi
fied in a wide range of habitats in the Neotropical region. The Amazon exhibits the
highest diversity of Cichlidae, with more than 300 identified species (Kullander 2003). In Neotropical cichlids, the diversity of morphological adaptations does not re
sult from variations in the diploid number because most species have 48 chromo
somes (Feldberg et al. 2003). However, the species of this family exhibit a dynamic
genome with variations in structure and karyotype composition and organization,
as demonstrated by the DNA sequencing and the physical chromosome mapping
of several repetitive DNA sequences, such as telomere sequences; retrotransposons
isolated from Xiphophorus maculatus Günther, 1866 (Rex1, Rex3, Rex6); retrotrans
poson isolated from Astronotus ocellatus Agassiz, 1831 similar to Rex3 (AoRex3); long
interspersed elements isolated from Astronotus ocellatus (AoLINE); retrotransposon
isolated from Cichla kelberi Kullander & Ferreira, 2006 (RCk); transposon isolated
from Caenorhabditis elegans Maupas, 1900 (Tc1); the 18S and 5S ribosomal gene se
quences; and U1 spliceosomal small nuclear RNA (U1 snRNAs) (Vicari et al. 2006,
Gross et al. 2009, Mazzuchelli and Martins 2009, Teixeira et al. 2009, Gross et al. 2010, Poletto et al. 2010, Cabral-de-Mello et al. 2012, Valente et al. 2011, Schnei
der et al. 2013a, Schneider et al. 2013b). Moreover, in this fish family, repetitive
DNAs, such as transposable elements, co-localize or are associated with ribosomal
DNAs, which suggests their roles in the duplication and dispersion of repetitive
rDNA sequences (Gross et al. 2010, Schneider et al. 2013a, Schneider et al. 2013b,
Nakajima et al. 2012). Repetitive DNA sequences display a high degree of polymorphism because of the
variation in the number of repetitive units, which results from a specific evolution
ary dynamics. Among these elements, microsatellites (or short tandem repeats) are
the most polymorphic and are formed of short sequences of one to six nucleotides
repeated in tandem throughout the DNA (Tautz and Renz 1984). Keywords y
Karyotype evolution, fluorescence in situ hybridization, repetitive DNA, genome organization Abstract Fish of the family Cichlidae are recognized as an excellent model for evolutionary studies because of their
morphological and behavioral adaptations to a wide diversity of explored ecological niches. In addition, the
family has a dynamic genome with variable structure, composition and karyotype organization. Microsatel
lites represent the most dynamic genomic component and a better understanding of their organization may
help clarify the role of repetitive DNA elements in the mechanisms of chromosomal evolution. Thus, in this
study, microsatellite sequences were mapped in the chromosomes of Cichla monoculus Agassiz, 1831, Ptero
phyllum scalare Schultze, 1823, and Symphysodon discus Heckel, 1840. Four microsatellites demonstrated
positive results in the genome of C. monoculus and S. discus, and five demonstrated positive results in the
genome of P. scalare. In most cases, the microsatellite was dispersed in the chromosome with conspicuous
markings in the centromeric or telomeric regions, which suggests that sequences contribute to chromosome
structure and may have played a role in the evolution of this fish family. The comparative genome mapping
data presented here provide novel information on the structure and organization of the repetitive DNA
region of the cichlid genome and contribute to a better understanding of this fish family’s genome. 596 Carlos Henrique Schneider et al. / Comparative Cytogenetics 9(4): 595–605 (2015) Introduction Because of their
supposed neutral evolution, these molecular markers have been widely used in popula
tion genetics, to identify taxonomic limits, and in hybridization and forensic studies
(Goldstein and Pollock 1997, Filcek et al. 2005, Racey et al. 2007, McCusker et
al. 2008). However, recent research has demonstrated that certain microsatellites are Chromosomal distribution of microsatellite repeats in Amazon cichlids genome... 597 functional and may affect gene regulation (Kashi and King 2006, Gemayel et al. 2010,
Sonay et al. 2015); evolution of orphan genes (Palmieri et al. 2014, Schlötterer 2015);
involved in chromosomal rearrangements (Kamali et al. 2011) and be involved with
increased or diminishes likelihood of disease related alleles size (Ananda et al. 2014;
Padeken et al. 2015).h The chromosomal mapping of microsatellite sequences has been little examined. This approach is primarily used to study the evolution of different sex-determining
chromosome systems (Li et al. 2010, Pokorná et al. 2011, Terencio et al. 2013). Con
sidering that microsatellites are the most dynamic genomic component, a better un
derstanding of their chromosomal organization is important for improving knowledge
regarding the role of repetitive DNA elements in the mechanisms of chromosomal
evolution and heterochromatin composition. p
Cichla monoculus Agassiz, 1831 has a karyotype with 2n = 48 subtelo/acrocentric
(st/a) chromosomes, described as basal for cichlids, and little heterochromatin. Al
though Pterophyllum scalare Schultze, 1823 also has 2n = 48 chromosomes, this species
differs in karyotype formula with meta/submetacentric (m/sm) chromosomes due to
chromosomal inversions and accumulation of heterochromatin in the pericentromeric
regions. The highest diploid number described for this group is found in species of the
genus Symphysodon Heckel, 1840, which has 2n = 60 chromosomes, as well as large
heterochromatic blocks (Schneider et al. 2013a). Thus, this study provides a physical
mapping of microsatellite sequences on the chromosomes of three Neotropical cichlid
fish species (C. monoculus, P. scalare, and S. discus), that occupy different phylogenetic
positions, and contributes to a better understanding of the chromosomal organization
and evolution of this fish family. Methods Specimens of C. monoculus (four males and four females), P. scalare (three males and
three females) were collected in Catalão Lake, confluence of the Negro/Solimões Riv
ers (3°09'47.44"S / 59°54'51.39"W) and S. discus (two males and two females) in
Negro River (0°56'06.43"S / 62°56'22.61"W). The specimens were caught in the wild
with sampling permission (ICMBio SISBIO 10609-1/2007). All of the individuals
were euthanatized with Eugenol (clove oil). Mitotic chromosomes were obtained from kidney cells using an air-drying proto
col (Bertollo et al. 1978). Eight microsatellites (Amado et al. 2008; Passos et al. 2010) were mapped using
fluorescence in situ hybridization (FISH) during the mitotic metaphase of C. monocu
lus, P. scalare, and S. discus (Table 1). The repetitive sequence probes were labeled with
digoxigenin-11-dUTP or biotin-16-dUTP (Dig-Nick Translation Mix and BioNick
Translation Mix; Roche) using nick translation reactions following the manufacturer’s
instructions. Anti-digoxigenin rhodamine (Roche) and streptavidin/Alexa Fluor 488
(Life Technologies) were used to detect the signal. FISH was performed on mitotic Carlos Henrique Schneider et al. / Comparative Cytogenetics 9(4): 595–605 (2015) 598 Table 1. Repetitive sequences hibridized to cichlid chromosomes. (+) positive hybridization signals de
tected; (-) absence of hibridization signals. chromosome spreads (Pinkel et al. 1986). The FISH was performed with high strin
gency (2.5 ng/µl of DNA, 50% deionized formamide, 10% dextran sulfate and 2xSSC
at 37 °C for 18 h). The chromosomes were counterstained with DAPI (2 µg/ml) in the
Vectashield mounting medium (Vector). p
q
p
y
g
tected; (-) absence of hibridization signals. Repeat motif
C. monoculus
P. scalare
S. discus
(CA)16
+
+
+
(AC)7
+
-
-
(GT)13
+
+
-
(GA)12
-
+
-
(GAATA)8
+
+
+
(GAGAA)12
-
+
-
(GT)9CA(GT)7CG(GT)19
-
-
+
(CT)14GT(CT)5(CG)2(CT)9
-
-
+ chromosome spreads (Pinkel et al. 1986). The FISH was performed with high strin
gency (2.5 ng/µl of DNA, 50% deionized formamide, 10% dextran sulfate and 2xSSC
at 37 °C for 18 h). The chromosomes were counterstained with DAPI (2 µg/ml) in the
Vectashield mounting medium (Vector). tected; ( ) absence of hibridization signals. Repeat motif
C. monoculus
P. scalare
S. discus
(CA)16
+
+
+
(AC)7
+
-
-
(GT)13
+
+
-
(GA)12
-
+
-
(GAATA)8
+
+
+
(GAGAA)12
-
+
-
(GT)9CA(GT)7CG(GT)19
-
-
+
(CT)14GT(CT)5(CG)2(CT)9
-
-
+ chromosome spreads (Pinkel et al. 1986). Methods The FISH was performed with high strin
gency (2.5 ng/µl of DNA, 50% deionized formamide, 10% dextran sulfate and 2xSSC
at 37 °C for 18 h). The chromosomes were counterstained with DAPI (2 µg/ml) in the
Vectashield mounting medium (Vector). Results Four microsatellites, among which three were dinucleotides and one was a pentanucle
otide, exhibited positive reactions in the genome of C. monoculus (Table 1). Hybridi
zations with the pentanucleotide microsatellite (GAATA)8 displayed dispersed signals
in all of the chromosomes. Moreover, conspicuous markings were observed in several
chromosome pairs. However, one chromosome pair did not exhibit any homology
with the probe (Fig. 1a). The microsatellite (CA)16 was distributed in all of the chromo
somes of C. monoculus, except for one pair. In most chromosomes, the microsatellite
displayed a dispersed distribution, and in several cases, the markings were conspicuous
(Fig. 1b). A dispersed pattern was observed after hybridization with the microsatellite
(GT)13, whereas a strong signal occurred in the telomeric, interstitial or centromeric
regions of the chromosomes (Fig. 1c). Conversely, hybridizations with the microsatel
lite (AC)7 resulted in only two positive chromosome pairs, one with markings in the
telomeric region of the short arm and in the interstitial region of the long arm and the
other with markings in both telomeric regions (Fig. 1d). Five microsatellites were mapped in the genome of P. scalare: three dinucleo
tides and two pentanucleotides (Table 1). The microsatellites (GAATA)8, (CA)16 and
(GT)13, which were dispersed in C. monoculus, exhibited clustered signals in P. scalare. The microsatellite (CA)16 was mapped on centromeric blocks of five chromosome pairs
(Fig. 1f), other signals were located in telomeric, centromeric and interstitial regions
of most chromosomes (Fig. 1e and 1g). Additionally, the microsatellites (GA)12 and
(GAGAA)12 exhibited similar patterns of conspicuous markings in the centromeres. However, in the case of (GAGAA)12, one chromosome pair did not display any mark
ings (Fig. 1h, i). Chromosomal distribution of microsatellite repeats in Amazon cichlids genome... 599 igure 1. Metaphase chromosomes of C. monoculus, P. scalare and S. discus hybridized with microsa
te sequences (a–m). Arrow shows no signs of hybridizations in a, b, h, i and chromosomes posi
or microsatellite in l, m. The probes detection was performed with streptavidin Alexa Fluor 488 (gre
r anti-digoxigenin rhodamine (red). Chromosomes were counterstained with DAPI. Scale bar: 10 µm Figure 1. Metaphase chromosomes of C. monoculus, P. scalare and S. discus hybridized with microsatel
lite sequences (a–m). Arrow shows no signs of hybridizations in a, b, h, i and chromosomes positive
for microsatellite in l, m. The probes detection was performed with streptavidin Alexa Fluor 488 (green)
or anti-digoxigenin rhodamine (red). Chromosomes were counterstained with DAPI. Discussion The repetitive regions of the genome typically do not undergo the selective pressure
that affects non-repetitive sequences, and most microsatellite sequences evolve neutral
ly and supposedly do not affect an individual phenotype (Schlötterer 2000). However,
recent studies have identified functional microsatellites that affect the physical aspect
of an individual (Kashi and King 2006, Gemayel et al. 2010, Padeken et al. 2015). These putative functional microsatellites are primarily located in or near gene regions,
and there is variation in the number of times that the motif is repeated, which is re
lated to the ability of the microsatellites to modify gene expression or change protein
sequences (Wren et al. 2000, Li et al. 2004, Vinces et al. 2009, Gemayel et al. 2010). In
addition to this functional aspect, repetitive DNA variants that include microsatellites
may serve as efficient agents for adaptive evolution (King and Kashi 2009). fi
In all classes of repetitive DNA, there appears to be a general trend of increased
matrix length throughout evolutionary time. Moreover, highly repetitive sequences
tend to accumulate in regions of low recombination, such as centromeres and telom
eres, whereas repetitive regions in euchromatin are much more susceptible to crossing-
over (Pathak and Ali 2012). Overall, the chromosome hybridization of microsatellites demonstrated contrast
ing patterns of abundance and localization of these sequences in the chromosomes of
C. monoculus, P. scalare and S. discus, which indicates that the repetitive sequences have
accumulated differently among the genomes. Although the three species exhibited a
wide distribution of microsatellites (GAATA)8 and (CA)16 in their genome, cluster
ing of these markers was observed in P. scalare, which represents a derived species in
the phylogeny of Cichlinae (Smith et al. 2008). Clustering of repetitive sequences in
derived species was also observed for transposable elements of this fish family (Gross
et al. 2009; Valente et al. 2011; Schneider et al. 2013b). The association between
microsatellites and the abundance of retrotransposable elements has been suggested
as a mechanism that may increase the number of microsatellites (Nadir et al. 1996). However, in several organisms, a relationship between the high density of transposable
elements and a high rate of microsatellites is not observed (Schlötterer 2000, 2015). Position of the microsatellite sequences mapped in this study was similar to that
observed for retroelements in the same species, with signals scattered throughout chro
mosomes and others clustered in terminal and centromeric regions (Schneider et al. 2013b). Results Scale bar: 10 µm. 600 Carlos Henrique Schneider et al. / Comparative Cytogenetics 9(4): 595–605 (2015) Four microsatellites were mapped in S. discus. In this species, the microsatellites
(GAATA)8 and (CA)16 exhibited patterns similar to that of C. monoculus with dispersed
signals and conspicuous markings in the centromeric region (Fig. 1j, 1k). Only one chro
mosome pair displayed positive results for microsatellites (GT)9CA(GT)7CG(GT)19 and
(CT)14GT(CT)5(CG)2(CT)9. The first was located in the telomeric region of the long
arm, and the latter was located in the centromeric and telomeric regions (Fig. 1l, m). Discussion This outcome suggests that these sequences may be involved in the structural Chromosomal distribution of microsatellite repeats in Amazon cichlids genome... 601 formation of the centromere and the telomere. Moreover, the microsatellites present
in the centromeric and subtelomeric regions differ among cichlid species, which rein
forces the importance of these sequences in the evolution of the different groups. p
qf
g
p
The centromere is an essential structure with several functional roles in the segre
gation of replicated chromosomes to daughter cells. These roles include genetic/epi
genetic marking and the assembly of the protein complex of the kinetochore during
the cell cycle, providing checkpoints to control mitosis, the release of sister-chromatid
cohesion, chromosome migration to the cellular poles and cytokinesis (Santaguida
and Musacchio 2009, Allshire and Karpen 2008, Perpelescu and Fukagawa 2011). Centromeres comprise long stretches of tandem repeats of satellite and microsatel
lite DNA, which are essential for binding with protein complexes (Verdaasdonk and
Bloom 2011, Kalitsis and Choo 2012). Centromeric DNA sequences typically present
high evolutionary rates and variation among species or chromosomes of the same spe
cies is common (Plohl et al. 2008). Thus, the centromeric region of P. scalare is rich in
(GAGAA)12 microsatellite sequences, but one chromosome pair does not display any
hybridization signals. As well, in C. monoculus and S. discus, the centromere region is
not rich in (GA)12 and (GAGAA)12. 12
12
Another chromosome region with a high evolutionary rate is the subterminal re
gion. Typically, this region is composed of different classes of repetitive DNA that
may help stabilize the terminal portion of the chromosomes because of the possibil
ity of amplifying these sequences even in the absence of telomerase (Jain et al. 2010,
Torres et al. 2011). In S. discus, the microsatellites (GT)9CA(GT)7CG(GT)19 and
(CT)14GT(CT)5(CG)2(CT)9 are present in the subterminal region of two chromosome
pairs, whereas these markers were not observed in the chromosomes of C. monoculus
and P. scalare. The same result was obtained for the microsatellite (AC)7, which was ob
served only in C. monoculus. The microsatellites (GAATA)8 and (GT)13 were observed
in the subterminal regions of C. monoculus and P. scalare but not in S. discus. In P. sca
lare, a conspicuous marking of the microsatellite (GT)13 was observed in the terminal
region of the largest chromosome pair, where the 18S ribosomal gene is located (Sch
neider et al. Discussion 2013a), indicating synteny between these two classes of repetitive DNA. Still, heterochromatin of the cichlids analyzed here was located in the centromeric
or pericentromeric regions in most of the chromosomes (Schneider et al. 2013). These
regions show positive signals of hybridization for different microsatellites analyzed, as
well as other repetitive elements (Schneider et al. 2013). The most common cellular
mechanism that prevents activation and expansion of repetitive elements is the forma
tion of heterochromatin over their sequences and three epigenetics pathways intercon
nected ensure the silencing of their elements: methylation of H3K9, DNA methyla
tion and the germ-line specific PIWI pathway (Padeken et al. 2015). gi
The regulation of the repetitive sequences is not yet clear and depends largely on new
technologies to clarify their function (Padeken et al. 2015), but the comparative map
ping data presented provide novel information on the structure and organization of the
repetitive region of the genome of cichlids and suggest that microsatellites contribute to
chromosome structure and may have played a role in the evolution of this fish family. 602 Carlos Henrique Schneider et al. / Comparative Cytogenetics 9(4): 595–605 (2015) Acknowledgments This study was supported by the Conselho Nacional de Pesquisa and Desenvolvimento
Tecnológico (CNPq—140816/2009-7 CHS scholarship), Instituto Nacional de Pes
quisas da Amazônia/Biologia de Água Doce e Pesca Interior (INPA/BADPI), Fundação
de Amparo de Pesquisas do Amazonas (PRONEX FAPEAM/CNPq 003/2009),
Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP) and the CNPq/
Universal (476292/2013-1). References Allshire RC, Karpen GH (2008) Epigenetic regulation of centromeric chromatin: old dogs,
new tricks? Nature Reviews Genetics 9: 923–937. doi: 10.1038/nrg2466 Amado MV, Hrbek T, Gravena W, Fantin C, Assunção EM, Astolfi-Filho S, Farias IP (2008)
Isolation and characterization of microsatellite markers for the ornamental discus fish Sym
physodon discus and cross-species amplification in other Heroini cichlid species. Molecular
Ecology Resources 8: 1451–1453. doi: 10.1111/j.1755-0998.2008.02200.x Ananda G, Hile SE, Breski A, Wang Y, Kelkar Y, Makova KD, Eckert KA (2014) Microsatel
lite interruptions stabilize primate genomes and exist as population-specific single nucleo
tide polymorphisms within individual human genomes. PLoS Genetics 10: e1004498. doi:
10.1371/journal.pgen.1004498 Bertollo LAC, Takahashi CS, Moreira-Filho O (1978) Cytotaxonomic considerations on Hoplias
lacerdae (Pisces, Erythrinidae). Brazilian Journal of Genetics 1: 103–120. Cabral-de-Mello DC, Valente GT, Nakajima RT, Martins C (2012) Genomic organization
and comparative chromosome mapping of the U1 snRNA gene in cichlid fish, with an
emphasis in Oreochromis niloticus. Chromosome Research 20: 279–292. doi: 10.1007/
s10577-011-9271-y Feldberg E, Porto JIR, Bertollo LAC (2003) Chromosomal changes and adaptation of cichlid
fishes during evolution. In: Val AL, Kapoor BG (Eds) Fish adaptation. Science Publishers,
Enfield, USA, 285–308. Filcek KB, Gilmore SA, Scribner KT, Jones ML (2005) Discriminating lamprey species using
multilocus microsatellite genotypes. North American Journal of Fisheries Management 25:
502–509. doi: 10.1577/M03-206.1 Gemayel R, Vinces MD, Legendre M, Verstrepen KJ (2010) Variable tandem repeats accelerate
evolution of coding and regulatory sequences. Annual Reviews of Genetics 44: 445–477. doi: 10.1146/annurev-genet-072610-155046 Goldstein DB, Pollock DD (1997) Lauching microsatellities. Jounal of Heredity 88: 335–342. doi: 10.1093/oxfordjournals.jhered.a023114 Gross MC, Schneider CH, Valente GT, Porto JIR, Martins C, Feldberg E (2009) Comparative
cytogenetic analysis of the genus Symphysodon (Discus Fishes, Cichlidae): chromosomal
characteristics of retrotransposons and minor ribosomal DNA. Cytogenetic and Genome
Research 127: 43–53. doi: 10.1159/000279443 Chromosomal distribution of microsatellite repeats in Amazon cichlids genome... 603 Gross MC, Schneider CH, Valente GT, Porto JIR, Martins C, Feldberg E (2010) Variability
of 18S rDNA locus among Symphysodon fishes: chromosomal rearrangements. Journal of
Fish Biology 76: 1117–1127. doi: 10.1111/j.1095-8649.2010.02550.x Jain D, Hebden AK, Nakamura TM, Miller KM, Cooper JP (2010) HAATI survivors replace
canonical telomeres with blocks of generic heterochromatin. Nature 467: 223–227. doi:
10.1038/nature09374 Kalitsis P, Choo KHA (2012) The evolutionary life cycle of the resilient centromere. Chromosoma
121: 327–340. References Cur
rent Opinion in Genetics & Development 31: 12–19. doi: 10.1016/j.gde.2015.03.009 j g
Palmieri N, Kosiol C, Schlötterer C (2014) The life cycle of Drosophila orphan genes. eLife 3:
1–21. doi: 10.7554/eLife.01311 Passos KB, Leão ASA, Oliveira DP, Farias IP, Hrbek T (2010) Polymorphic microsatellite
markers for the overexploited Amazonian fish, Semaprochilodus insignis (Jardine and
Schomburgk 1841). Conservation Genetics Resources 2: 231–234. doi: 10.1007/s12686-
010-9245-y Pathak D, Ali S (2012) Repetitive DNA: A tool to explore animal genomes/ transcriptomes. In: Germana M, Petrera F (Eds) Functional genomics. InTech, Published, 155–180. doi:
10.5772/48259 Perpelescu M, Fukagawa T (2011) The ABCs of CENPs. Chromosoma 120: 425–446. doi:
10.1007/s00412-011-0330-0 Pinkel D, Straume T, Gray JW (1986) Cytogenetic analysis using quantitative, high-sensitiv
ity, fluorescence hybridization. Proceedings of the National Academy of Sciences of the
USA 83: 2934–2938. doi: 10.1073/pnas.83.9.2934 Plohl M, Mesltrovin N, Bruvo B, Ugarkovic D (2008) Similarity of structural features and evo
lution of satellite DNAs from Palorus subdepressus (Coleoptera) and related species. Journal
of Molecular Evolution 46: 234–239. doi: 10.1007/PL00006298 Pokorná M, Kratochvíl L, Kejnovský E (2011) Microsatellite distribution on sex chromosomes
at different stages of heteromorphism and heterochromatinization in two lizard species
(Squamata: Eublepharidae: Coleonyx elegans and Lacertidae: Eremias velox). BMC Genetics
12: 90. doi: 10.1186/1471-2156-12-90 Poletto AB, Ferreira IA, Cabral-de-Mello DC, Nakajima RT, Mazzuchelli J, Ribeiro HB, Ve
nere PC, Nirchio M, Kocher TD, Martins C (2010) Chromosome differentiation patterns
during cichlid fish evolution. BMC Genetics 11: 50. doi: 10.1186/1471-2156-11-50 Racey PA, Barratt EM, Burland TM, Deaville R, Gotelli D, Jones G, Piertney SB (2007)
Microsatellite DNA polymorphism confirms reproductive isolation and reveals differences
in population genetic structure of cryptic pipistrelle bat species. Biological Journal of the
Linnean Society 90: 539–550. doi: 10.1111/j.1095-8312.2007.00746.x Santaguida S, Musacchio A (2009) The life and miracles of kinetochores. EMBO Journal 28:
2511–2531. doi: 10.1038/emboj.2009.173 Schliewen UK, Tautz D, Pääbo S (1994) Sympatric speciation suggested by monophyly of
crater lake cichlids. Nature 368: 629–632. doi: 10.1038/368629a0 Schlötterer C (2000) Evolutionary dynamics of microsatellite DNA. Chromosoma 109: 365–371. doi: 10.1007/s004120000089 Schlötterer C (2015) Genes from scratch – the evolutionary fate of de novo genes. Trends in
Genetics 31: 215–219. doi: 10.1016/j.tig.2015.02.007 Schneider CH, Gross MC, Terencio ML, Artoni RF, Vicari MR, Martins C, Feldberg E
(2013a) Chromosomal evolution of neotropical cichlids: the role of repetitive DNA se
quences in the organization and structure of karyotype. Reviews in Fish Biology and Fish
eries 23: 201–214. References doi: 10.1007/s00412-012-0369-6 Kamali M, Sharakhova MV, Baricheva E, Karagodin E, Tu Z, Sharakhov IV (2011) An inte
grated chromosome map of microsatellite markers and inversion breakpoints for an Asian
malaria mosquito, Anopheles stephensi. Journal of Heredity 102: 719–726. doi: 10.1093/
jhered/esr072 Kashi Y, King DG (2006) Simple sequence repeats as advantageous mutators in evolution. Trends in Genetics 22: 253–259. doi: 10.1016/j.tig.2006.03.005 King D, Kashi GY (2009) Heretical DNA sequences? Science 326: 229–230. doi: 10.1126/
science.326_229 Kocher TD (2004) Adaptive evolution and explosive speciation: the cichlid fish model. Nature
Reviews Genetics 5: 288–298. doi: 10.1038/nrg1316 Kullander SO (1998) A phylogeny and classification of the South America Cichlidae (Teleostei:
Perciformes). In: Malabarba LR, Reis RE, Vari RP, Lucena ZMS, Lucena CAS (Eds) Phylogeny
and classification of neotropical fishes. EdiPuc, Porto Alegre, 461–498. ii
Kullander SO (2003) Family Cichlidae. In: Reis RE, Kullander SO, Ferraris JrCJ (Eds) Check
List of the Fresh-water Fishes of South and Central America. EdiPuc, Porto Alegre, 605–654. Li YC, Korol AB, Fahima T, Nevo E (2004) Microsatellites within genes: structure, function, and
evolution. Molecular Biology and Evolution 21: 991–1007. doi: 10.1093/molbev/msh073 Li J, Phillips RB, Harwood AS, Koop BF, Davidson WS (2010) Identification of the sex chro
mosomes of brown trout (Salmo trutta) and their comparison with the corresponding
chromosomes in Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss). Cytogenetic and Genome Research 133: 25–33. doi: 10.1159/000323410 Lowe-McConnell RH (1991) Ecology of cichlids in South American and African waters, ex
cluding the African Great Lakes. In: Keenleyside MHA (Ed.) Cichlid Fishes – Behavior,
Ecology and Evolution. Chapman and Hall, London, 60–85. Mazzuchelli J, Martins C (2009) Genomic organization of repetitive DNAs in the cichlid fish
Astronotus ocellatus. Genetica 136: 461–469. doi: 10.1007/s10709-008-9346-7 McCusker MR, Paterson IG, Bentzen P (2008) Microsatellite markers discriminate three spe
cies of North Atlantic wolffishes (Anarhichas spp.). Journal of Fish Biology 72: 375–385. doi: 10.1111/j.1095-8649.2007.01701.x Nadir E, Margalit HT, Gallily T, Ben-Sasson SA (1996) Microsatellite spreading in the hu
man genome: Evolutionary mechanisms and structural implications. Proceedings of the
National Academy of Sciences of the USA 93: 6470–6475. doi: 10.1073/pnas.93.13.6470 Nakajima RT, Cabral-de-Mello DC, Valente GT, Venere PC, Martins C (2012) Evolutionary
dynamics of rRNA gene clusters in cichlid fish. BMC Evolutionary Biology 12: 198. doi:
10.1186/1471-2148-12-198 604 Carlos Henrique Schneider et al. / Comparative Cytogenetics 9(4): 595–605 (2015) Padeken J, Zeller P, Gasser SM (2015) Repeat DNA in genome organization and stability. References doi: 10.1007/s11160-012-9285-3 Chromosomal distribution of microsatellite repeats in Amazon cichlids genome... 605 Schneider CH, Gross MC, Terencio ML, Carmo EJ, Martins C, Feldberg E (2013b) Evolu
tionary dynamics of retrotransposable elements Rex1, Rex3 and Rex6 in Neotropical cich
lids genome. BMC Evolutionary Biology 13: 152. doi: 10.1186/1471-2148-13-152 Seehausen O (2006) African cichlid fish: a model system in adaptive radiation research. Pro
ceedings of the Royal Society B: Biological Sciences 273: 1987–1998. doi: 10.1098/
rspb.2006.3539 Smith WL, Chakrabarty P, Sparks JS (2008) Phylogeny, taxonomy, and evolution of Neotropi
cal cichlids (Teleostei: Cichlidae: Cichlinae). Cladistics 24: 625–641. doi: 10.1111/j.1096-
0031.2008.00210.x Sonay TB, Carvalho T, Robinson MD, Greminger MP, Krützen M, Comas D, Highnam G,
Mittelman D, Sharp A, Bonet TM, Wagner A (2015) Tandem repeat variation in human
and great ape populations and its impact on gene expression divergence. bioRxiv. doi:
10.1101/015784 Tautz D, Renz M (1984) Simple sequences are ubiquitous repetitive components of eukaryotic
genomes. Nucleic Acids Research 25: 4127–4138. doi: 10.1093/nar/12.10.4127 Teixeira WG, Ferreira IA, Cabral-de-Mello DC, Mazzuchelli J, Valente GT, Pinhal D, Poletto
AB, Venere PC, Martins C (2009) Organization of repeated DNA elements in the genome
of the cichlid fish Cichla kelberi and its contributions to the knowledge of fish genomes. Cytogenetic and Genome Research 125: 224–234. doi: 10.1159/000230006 Terencio ML, Schneider CH, Gross MC, Vicari MR, Farias IP, Passos KB, Feldberg E
(2013) Evolutionary dynamics of repetitive DNA and sex chromosomes: Semaprochilodus
(Characiformes, Prochilodontidae) as a model. Sexual Development 7: 325–333. doi:
10.1159/000356691 Torres GA, Gong Z, Iovene M, Hirsch CD, Buell CR, Bryan GJ, Novák P, Macas J, Jiang J
(2011) Organization and evolution of subtelomeric satellite repeats in the potato genome. Genetics 1: 85–92. doi: 10.1534/g3.111.000125 Valente GT, Mazzuchelli J, Ferreira IA, Poletto AB, Fantinatti BEA, Martins C (2011) Cytoge
netic mapping of the retroelements Rex1, Rex3 and Rex6 among cichlid fish: new insights
on the chromosomal distribution of transposable elements. Cytogenetic and Genome Re
search 133: 34–42. doi: 10.1159/000322888 Verdaasdonk JS, Bloom K (2011) Centromeres: unique chromatin structures that drive chromosome
segregation. Nature Reviews Molecular Cell Biology 12: 320–332. doi: 10.1038/nrm3107 Vicari MR, Artoni RF, Moreira-Filho O, Bertollo LAC (2006) Basic and molecular cytogenet
ics in freshwater Cichlidae (Osteichthyes, Perciformes). Karyotypic conservationism and
divergence. Caryologia 59: 260–266. doi: 10.1080/00087114.2006.10797924 Vinces M, Legendre M, Caldara M, Hagihara M, Verstrepen K (2009) Unstable tandem re
peats in promoters confer transcriptional evolve ability. Science 324: 1213–1216. References doi:
10.1126/science.1170097 Wren J, Forgacs E, Fondon J, Pertsemlidis A, Cheng S, Gallardo T, Williams R, Shohet R,
Minna J, Garner H (2000) Repeat polymorphisms within gene regions: phenotypic and
evolutionary implications. American Journal of Human Genetics 67: 345–356. doi:
10.1086/303013
|
https://openalex.org/W4394983815
|
https://azramedia-indonesia.azramediaindonesia.com/index.php/Kapalamada/article/download/170/147
|
Indonesian
| null |
PENANAMAN NILAI-NILAI KARAKTER PADA ANAK USIA DINI MELALUI MEDIA AUDIO VISUAL ANIMASI KARTUN BING BANI “KUCING”
| null | 2,022
|
cc-by
| 3,042
|
Jurnal Multidisipliner Kapalamada
e-ISSN : 2828 – 4194 (Online)
Volume : 01, Nomor : 02, Juni 2022 Abstrak Penelitien ini bertujuan untuk mendeskripsikan penanaman nilai-nilai
karakter pada anak usia dini melalui animasi kartun bing bani yang
meliputi penanaman sifat disiplin, jujur, bertanggung jawab, saling
menyayangi, religious, pemberani, dan pantang menyerah. Jenis
penelitian ini merupakan pendekatan kualitatif dengan teknik
pengumpulan data dan sumber datanya diperoleh langsung dari
animasi kartun bing bani. Hasil dari penelitian ini adalah penanaman
nilai-nilai karakter pada anak usia dini yang dilaksanakan melalui
media audio visual yang diharapkan dapat memberikan pengaruh
langsung terhadap anak mengenai berbagai karakter yang ada di
dalam animasi kartun tersebut. Solusi yang bias diberikan guru
terhadap peserta didik agar anak-anak tidak ketergantungan
menonton animasi kartun tersebut guru harus memberikan batasan
kepa anak saat menonton animasi tersebut dan guru juga harus
memberikan bimbingan, pendampingan, dan pembiasaan kepada
anak. e-ISSN : 2828 – 4194 (Online
Volume : 01, Nomor : 01, Maret 202
PENANAMAN NILAI-NILAI KARAKTER PADA ANAK USIA DINI
MELALUI MEDIA AUDIO VISUAL ANIMASI KARTUN
BING BANI “KUCING” Amanda Puspa Sapitri*a Fuaddilah Ali Sofyanb
a Pendidikan Islam Anak Usia Dini, Universitas Islam Negeri Raden Fatah Palembang
b Pendidikan Islam Anak Usia Dini, Universitas Islam Negeri Raden Fatah Palembang
Email: amandapuspasapitri@gmail.com, fuadpgmi_uin@radenfatah.ac.id Amanda Puspa Sapitri*a Fuaddilah Ali Sofyanb
a Pendidikan Islam Anak Usia Dini, Universitas Islam Negeri Raden Fatah Palembang
b Pendidikan Islam Anak Usia Dini, Universitas Islam Negeri Raden Fatah Palembang
Email: amandapuspasapitri@gmail.com, fuadpgmi_uin@radenfatah.ac.id ARTICLE HISTORY
Received:
01 Mei 2022
Revised
07 Mei 2022
Accepted:
25 Mei 2022
Online available:
10 Juni 2022
Keywords :
Nilai Karakter, Audio
Visual
Character value, Audio
Visual
*Correspondence:
Name : Amanda Puspa
Sapitri
E-mail: amandapuspa
sapitri@gmail.com PENDAHULUAN Dengan berkembangnya teknologi saat ini, apapun dan siapapun akan
dekat dengan adanya internet. Tidak hanya orang dewasa saat ini anak-anak juga
dapat mengakses internet, ditambah dengan adanya berbagai aplikasi yang
mendukung anak untuk melihat ataupun menonton apa yang diinginkan. Tidak
dipungkira lagi bahwa saat ini televise sudah tidak digemari anak-anak untuk
menonton berbagai animasi kartun, anak lebih memilih dan tertarik menonton
animasi kartun melalui aplikasi YouTube. YouTube merupakan situs video yang
menyediakan berbagai informasi berupa gambar bergerak dan bias diandalkan. Situs ini dapat dikatakan situs audio visual yang dapat memberikan edukasi pada
anak jika digunakan dengan benar (Baskoro, 2009). Terdapat berbagai macam tayangan yang ada di youtube mulai dari yang
mendidik bahkan yang tidak mendidik juga ada di aplikasi ini, salah satu
diantaranya yang sering di tonton oleh anak-anak itu ialah video animasi kartun
baik itu kartun yang berasal dari Negara sendiri maupun Negara luar semuanya
ada di youtube.Menurut Departement Pendidikan Nasional dalam Kamus Besar
Bahasa Indonesia animasi adalah rangkaian tulisan atau gambaran yang
digerakan
secara
mekanis
elektronis
sehingga
tampak
dilayar
menjadi
gerak.Animasi ini termasuk kedalam media audio visual yang dimana media
audiovisual ini dapat menyajikan gambar dan suara secara bersamaan sehingga
dapat merangsang anak pada saat melihatnya. Dalam penelitian ini peneliti menjadikan animasi kartun bing bani sebagai
media dalam menanamkan nilai-nilai karakter pada anak. Dimana saat ini nilai
karkter pada anak sudah sedikit menghilang dan tidak lagi dijadikan sebagai
kepentingan, untuk itu kita sebagai orang dewasa sangat perlu menanamkan nilai-
nilai karakter pada anak sejak dini agar nantinya anak yang tumbuh dan
berkembang tersebut memiliki nilai karakter yang dapat menjadikannya cerminan
dimasyarakat. Menurut Lickonal dalam suyadi pendidikan karakter memiliki tiga pokok
yakni mengetahui kebaikan (knowing the good), mencintai kebaikan (desiring the
good), dan melakukan kebaikan (doing the good) (Suyadi, 2013). Dimana dengan
tertanamnya ketiga pokok tersebut dalam diri anak akan menjadikan anak lebih
memahami nilai karakter yang ada dan tumbuh pada dirinya tersebut. Tujuan utama pendidikan karakter adalah mempasilitasi dan menguatkan
dalam mengembangkan nilai-nilai karakter sehingga dapat mewujudkan prilaku
anak yang baik dan sopan, baik dalam proses sekolah maupun proses diluar
sekolah (Kesuma, 2012). Tidak hanya itu tujuan dari penanaman nilai karakter
melali media audiovisual animasi bing bani diharapkan bahwa anak dapat memilih
kartun yang sesuai dengan usianya dan tentunya animasi kartun yang memiliki
pesan moral di setiap kejadiannya. Abstract This study aims to describe the cultivation of character values in early
childhood through the cartoon animation of Bing Bani which includes
the cultivation of discipline, honesty, responsibility, mutual love,
religious, courageous, and unyielding. This type of research is a
qualitative approach with data collection techniques and the data
source is obtained directly from the Bing Bani cartoon animation. The
result of this research is the inculcation of character values in early
childhood which is carried out through audio-visual media which is
expected to have a direct influence on children regarding the various
characters in the animated cartoon. The solution that teachers can
give to students so that children are not dependent on watching
cartoon animations, the teacher must provide limits to children when
watching the animation and the teacher must also provide guidance,
assistance, and habituation to children. 187 Amanda 1(2), 187-194 Jurnal Multidisipliner Kapalamada PENDAHULUAN Melalui tontonan animasi kartun bing bani ini diharapkan dapat
memberikan makna positif bagi setiap anak yang menontnya sehingga dapat
bertingkah laku yang baik dan sopan pada saat anak berada di lingkungan sosial, 188 Jurnal Multidisipliner Kapalamada Amanda 1(2), 187-194 tingkah laku anak merupakan pilar yang menjadi cerminan terhadap apa yang
didapat anak setelah menonton kartun animasi tersebut. Anak harus diupayakan
untuk memiliki kesadaran terhadap nilai karakter yang dimilikinya seperti anak
dapat bersifat baik, jujur, disiplin, menghargai orang lain, dan entunya memiliki
moral sebagai keteladanan. Melalui kebiasaan anak saat ini peneliti bertujuan untuk melakukan
penelitian terhadap anak melalui animasi bing bani. Apakah animasi bing bani ini
dapat memberikan pengaruh kepada nilai-nilai karakter yang dimiliki anak. Nilai-
nilai karakter yang ditanamkan pada anak sejak usia dini merupakan fondasi
penting bagi terbentuknya tatanan kehidupan masyarakat yang beradab dan
berakhlak mulia serta memiliki sifat yang menguntungkan bagi orang lain (Revell,
2007). Dalam penelitian ini peneliti menggunakan penelitian kualitatif agar dapat
menciptakan fenomena-fenomena baru dari fenomena sebelumnya terkait animasi
kartun.Penelitian kualitatif adalah penelitian yang ditujukan untuk mendeskripsikan
dan menganalisis fenomena-fenomena, peristiwa, aktivitas sosial secara alamiah
(Sutama, 2012).Sementara menurut Moleong metodelogi kualitatif sebagai
prosedur penelitian yang mendeskripsikan data berupa kata-kata tertulis atau lisan
dari orang-orang dan prilaku yang dapat diamati (Moleong, 2014). Penelitian ini dilakukan dengan pengamatan langsung melalui cara
meonton animasi kartun bing bani, dengan mengamati bagaimana karakter setiap
tokoh yang ada dalam animasi tersebut. Kemudian mengamati setiap prilaku dan
kejadian-kejadian yang terdapat di dalam alur cerita animasi tersebut sehingga
dapat memberikan arahan bagaimana cara menerapkan nilai-nilai karakter yang
ada pada animasi kartun bing bani tersebut. Dengan dilakukannya penelitian ini diharapkan bahwa media audio visual
dapat berpengaruh dalam menanamkan karakter pada anak usia dini. Menurut
Barbara dalamyang dikutip oleh fitria mengemukakan bahwa media audiovisual
adalah cara memproduksi dan menyampaikan bahan dengan menggunakan
peralatan mekanis dan elektronis untuk menyajikan pesan-pesan melalui gambar
dan suara (Fristiani, 2016). Oleh karena itu melalui media ini akan dapat
mempermudah guru dalam memberikan atau menanamkan nilai-nilai karakter
pada anak. HASIL DAN PEMBAHASAN Dalam animasi bing bani ini terdapat beberapa katakter diantaranya
seekor kelinci yang berwarna hitam, panda, semut,kelinci bewarna putih, dan
seekor tikus. Pada bagian awal animasi ini diperlihatkan bahwa seekor kelinci
bewarna hitam ini sedang berlari bersama teman-temannya namun saat berlari
kelinci hitam ini terjatuh, dan semua teman-temannya bertanya apakah terjadi
sesuatu dan berkata apakah kau baik-baik saja bing. Dari sini dapat kita lihat
bahwa yang dilakukan teman-teman bing merupakan salah satu cara dalam
menanamkan niliai karakter pada anak. Penanaman secara etimologi berasal dari
kata tanam yang berarti benih, yang semakin jelas dengan mendapat imbuhan me-
kan menjadi menanamkan yang memiliki arti menaburkan ajaran, paham, dan lain
sebagainya.Secara etimologi, istilah karakter berasal dari bahasa Latin character,
yang berarti tabiat, tabiat, sifat-sifat kejiwaan, budi pekerti, kepribadian serta
akhlak. Penanaman nilai-nilai karakter dan pembentukan kepribadian yang berbudi
luhur pada peserta didik sejak usia dini merupakan usaha yang strategis dan tepat
dilakukan agar terbentuk perilaku yang bermoral dan berbudi luhur pada anak,
terutama pada anak sejak usia dini yang sangat membutuhkan perhatian dan
bimbingan yang intensif dari berbagai pihak (Subianto, 2013). Animasi bing bani yang berjudul “Kucing”, di saat bing dan temannya flod
sedang berjalan bani melihat seekor kicing yang bernama Arlo. Bani dan flod
menyapa arlo kemudian arlo mendekat kepada bing dan sepertinya kucing ini
menyukai bing. Bing dan flod asik bermain dengan arlo. Melalui karakter bing
dapat kita lihat bahwa mencintai dan menyayangi antar sesame itu merupakan hal
yang sangat baik. Dengan adanya anaimasi yang menunjukan rasa saya antara
sesame ini dapat memberikan pengaruh yang baik dalam menumbuhkan nilai
karakter pada anak.Terdapat beberapa metode untuk menanamkan nilai-nilai
karakter pada anak yaitu diantaranya : 1. Metode pembiasaan adalah membiasakan anak untuk melakukan sikap dan
perilaku yang mengandung nilai-nilai karakter yang mulia secara berulang-
ulang dan terus menerus sampai sikap/perilaku tersebut tertanam kuat dalam
diri .anak dan menjadi sesuatu yang biasa dilakukan oleh anak secara sadar
tanpa paksaan dari siapapun karena mengetahui, mencintai dan merasakan
bahwa sikap/perilaku tersebut merupakan suatu kebaikan yang harus
dilakukan. 2. Metode peneladanan mengharuskan guru memberikan contoh suatu sikap dan
perilaku yang mengandung nilai-nilai karakter pada anak. 2. Metode peneladanan mengharuskan guru memberikan contoh suatu sikap dan
perilaku yang mengandung nilai-nilai karakter pada anak. 3. Metode cerita adalah guru menceritakan suatu kisah, baik itu kisah nyata
maupun berupa dongeng yang mengandung nilai-nilai karakter dan agama di
dalamnya. Lewat suatu cerita, guru dapat menyisipkan pesan-pesan moral dan
agama pada anak. METODE PENELITIAN Penelitian yang peneliti lakukan ini merupakan jenis penelitian kualitatif,
dimana data penelitian yang digunakan berupa kata-kata, gambar dan bukan
angka. Penelitian kualitatif ini bertujuan untuk memahami fenomena tentang apa
yang dirasakan oleh subjek penelitian seperti perilaku, persepsi,motivasi, serta
tindakan dan lain sebagainya, secara keseluruhan baik dalam bentuk kata-kata
maupun bahasa, pada suatu konteks yang alamiyah dengan memanfaatkan
berbagai metode ilmiah (Sugiyono, 2013). Teknik pengumpulan data dalam
penelitian ini melalui pengamatan langsung terhadap kartun animasi Bing Bani, 189 Jurnal Multidisipliner Kapalamada Amanda 1(2), 187-194 dimana sumber data yang diperoleh melalui internet dengan menggunakan
aplikasi atau situs yang bernama YouTube. HASIL DAN PEMBAHASAN Yang terpenting adalah guru harus mampu mengemas dan 190 Jurnal Multidisipliner Kapalamada Amanda 1(2), 187-194 menyampaikan cerita tersebut dengan cara yang menarik dan menggunakan
bahasa yang sederhana dan jelas sehingga dapat menarik perhatian anak dan
mudah dimengerti maknanya oleh mereka. menyampaikan cerita tersebut dengan cara yang menarik dan menggunakan
bahasa yang sederhana dan jelas sehingga dapat menarik perhatian anak dan
mudah dimengerti maknanya oleh mereka. 4. Metode demonstrasi adalah guru memperagakan atau mempertunjukkan
perilaku nilai-nilai karakter yang hendak ditanamkan pada anak dihadapan
mereka sendiri. Selain bermanfaat memberikan ilustrasi dalam menjelaskan
suatu informasi, metode ini juga dapat membantu meningkatkan daya pikir
mereka dalam mengenali, mengingat dan berpikir evaluatif. Apa bila ada yang
keliru, maka akan dikoreksi oleh guru tersebut dan memberi penjelasan-
penjelasan sehingga anak tidak lagi melakukan kesalahan di dalam mengambil
kesimpulan. 5. Metode bermain digunakan karena anak-anak yang masih dalam kategori usia
dini senang bermain dan banyak menghabiskan waktunya dengan permainan. Bermain akan melatih anak untuk menggunakan kemampuan kognitifnya dan
dapat mengembangkan kreatifitasnya dengan melakukan eksplorasi. 6. Metode karyawisata adalah guru mengajak anak untuk mengunjungi suatu
tempat yang mengandung nilai-nilai pendidikannya, kemudian melakukan
observasi terhadap apa yang terdapat pada tempat tersebut. 6. Metode karyawisata adalah guru mengajak anak untuk mengunjungi suatu
tempat yang mengandung nilai-nilai pendidikannya, kemudian melakukan
observasi terhadap apa yang terdapat pada tempat tersebut. Pendidikan karakter dan penanaman nilai-nilai karakter sangatlah penting
diberikan pada anak sejak usia dini, karena pada usia dini merupakan awal yang
paling baik dan mudah untuk membentuk karakter anak (Zelvi, 2017). Pada sin selanjutnya bing yang sedang asik bermain bersama arlo, tiba-
tiba arlo pergi meninggalkan bing namun bing dan flod mengejar arlo yang sedang
berlari. Pada saat yang bersamaan bing dan flod melihat seekor kucing yang
tengah tertidur diatas kursi dan ternyata itu arlo, karena kucing tersebut sedang
tertidur bing dan arlo segera pergi meninggalkan kucing tersebut. Sifat yang
ditunjukan oleh bing ini merupakan sikap yang mencerminkan bagaimana
melampiaskan nilai kebaikan dalam bentuk tindakan atau tingkah laku yang baik
(Aulina, 2013). Pada saat yang bersamaan bing melihat ada seekor kucing yang sedang
berada di bawah tangga, kucing itu terlihat seperti Arlo. Kemudian bing mengajak
Flod untuk mendekatinya namun sepertinya kucing baru ini tidak menyukai bing
namun bing sangat ingin bermain bersamanya tetapi kucing baru ini terus pergi
meninggalkan bing dan flod. HASIL DAN PEMBAHASAN Disini terlihat bahwa bing tidak ingin menyerah dalam
melakukan apapun sekalipun dia disakiti oleh kucing tersebut, nilai karakter yang
terlihat disini bahwa sifat pantang menyerah yang ditunjukan bing sangat
mempengaruhi karakter anak, selain sifat pantang menyerah nilai-nilai karakter
pada anak juga dapat dilihat dari berbagai sifat lainnya yaitu karakter jujur,
toleransi, disiplin, kreatif, mandiri, demokratis, rasa ingin tahu, semangat
kebangsaan, cinta tanah air, menghargai prestasi, bersahabat atau berkomuniksi,
peduli lingkungan dan sosial dan rasa tanggung jawab yang besar (Permana,
1961). 191 Jurnal Multidisipliner Kapalamada Amanda 1(2), 187-194 Bing mengetahui bahwa kucing itu bukan lah arlo, ternyata kucing itu
hanyalah mirip dengan arlo.Ternyata kucing itu adalah Miten kucing dari
tetangganya. Setelah bing mengetahui semuanya bahwa Miten bukanlah kucing
yang jahat seperti dugaannya sebelumnya. Miten merasa bahwa bing akan
berbuat jahat terhadap anak-anaknya oleh karena itu miten berprilaku jahat dan
marah terhadap bing, pada akhirnya bing dan miten juga berteman dengan
baik.Dari kejadian ini kita dapat belajar bahwa kita harus selalu bersikap baik
terhadap siapapun dan apapun. Nilai-nilai karakter yang paling mendasar dan
utama yang harus ditanamkan serta dikembangkan pda peserta didik yang
meliputi : kejujuran, kesopanan, keikhlasan hati, empati, tanggung jawab,
kemandirian, kedisiplinan, keberanian, kasih saying, pengendalian diri, gotong
royong dan semangat serta bersungguh-sunggu dalam belajar dan berkarya
(Megawangi, 2004). Melalui animasi kartun bing bani ini terdapat beberapa cara dalam
membangun atau menumbuhkan nilai karakter pada anak, salah satunya dengan
mengajak anak menonton animasi tersebut, dengan enggunakan media audio
visualanakakan merasa tertarik dan tidak bosan cara ini juga dapat
mempermudah guru dalam meberikan stimulasi kepada anak. Media audio visual
merupakan rangkaian gambar elektronis yang disertai oleh unsur suara audio juga
mempunyai unsur gambar yang dituangkan melalui pita video, Rangkaian gambar
elektronis tersebutkemudian diputar dengan suatu alat yaitu video cassette
recorder atau video player (Hafied, 2006). Pemilihan media Audio Visual sebagai media dalam menanamkan nilai
karakter pada anak usia dini diharapkan agar mempermudah pengenalan nilai
karakter yang sesungguhnya pada anak. kartun animansi Bing dengan judul
kucing mengajarkan kepada ank bahwa sikap perduli terhadap sesama maupun
terhadap hewan sangat diperlukan, melalui kartun ini bukah hanya sifat peduli
yang harus di tanamkan namun banyak sekali sifat yang dapat menumbuhkan
karakter pada anak seperti sifat saling menyayangi, mudah bergaul, jujur dan rasa
ingin tahu terhadap apa pun. maka diharapkan melalui media audio visual ini
penanaman nilai karakter pada anak dapat lebih mudah dilakukan. KESIMPULAN Penanaman Nilai-nilai karakter pada anak usia dini sangat diperlukan
karena melalui penanaman karakter diharapkan dapat memberikan anak
kesempatan dalam mengembangkan dirinya. Penanaman karakter pada anak usia
dini dapat menggunakan beberapa metode dimana setiap metode mempunyai
cara tersendiri dalam memberikan stimulasi kepada anak tentang penanaman nilai
karakter pada anak. Nah penanaman nilai karakter ini juga dapat melalui berbagai
media pembelajaran, salah satunya yaitu dengan menggunakan media audio
visual.Media audio visual ini sangat efektif karena melalui suara dan gambar anak
lebih mudah memahami nilai karakter yang ada di kartun Animasi Bing Bani yang
telah ditontonnya. Dalam animasi kartun bing bani sangat banyak nilai positifny, 192 Jurnal Multidisipliner Kapalamada Amanda 1(2), 187-194 dan juga banyak nilai moral di setiap karakternya sehingga melalui animasi ini
dapat diharapkan memberikan kesempatan bagi guru atau orang tua dalam
menanamkan nilai karakter pada anak. Oleh karena itu penanaman nilai-nilai
karakter melalui media audio visual ini diharapkan membantu para guru atau orang
tua dalam melatihsetiap sifat yang dimiliki anak. REKOMENDASI Peneliti sangat merekomendasikan animasi kartun bing bani ini untuk
ditonton dan dilihat anak usia dini, karena didalam animasi tersebut banyak
mengandung nilai-nilai karakter sehingga melalui animasi tersebut dapat
memberikan stimulasi secara langsung kepada anak. Dan saya sangat
merekomendasikan animasi kartun bing bani ini untuk para orang tua dan lembaga
PAUD agar memberikan tontonan yang beredukasi serta mengandung nilai-nilai
yang baik untuk anak usia dini. DAFTAR PUSTAKA
Aulina, C, N. 2013. Penanaman disiplin Pada Anak Usia Dini. Jurnal Pedagogik,
Vol 1, No 2. DAFTAR PUSTAKA
Aulina, C, N. 2013. Penanaman disiplin Pada Anak Usia Dini. Jurnal Pedagogik,
Vol 1, No 2. Baskoro, Adi. 2009. Panduan Praktis searching di Internet. Jakarta: PT Trans
Media Baskoro, Adi. 2009. Panduan Praktis searching di Internet. Jakarta: PT Trans
Media Dharma Kesuma.2012. Pendidikan Karakter Kajian Teori dan Praktik di Sekolah. Bandung: PT Rwmaja Rosadakarya. Dharma Kesuma.2012. Pendidikan Karakter Kajian Teori dan Praktik di Sekolah. Bandung: PT Rwmaja Rosadakarya. Felly Permanan, 1961.Moral Education.PT gelora aksara Pratama. Fristiana Iriana. 2016, Pengembangan Kurikulum teori, Konsep Dan aplikasi. Yogyakarta: Dua satria Obset. Hafied, Cangara, 2006. Pengantar Ilmu Komunikasi, Jakarta: PR Raja Grafindo. https://youtubbe.com/watch?v=bMR7rg1jNgU&feature=share Megawangi, R. 2004.Pendidikan Karakter Solusi Yang Tepat Untuk Membangun
Bangsa. Jakarta: Indonesia Haritage Foundation. Mischo, C, et all. (2014). Does Early Childhood Teacher Education Affect Students
'Cognitive Orientations? The Effect of Different Education Tracks in
Teacher Education on Prospective Early Childhood Teachers' Cognitive
Orientations in Germany. The Journal of Education and Training Studies,
Vol 2, No 1. Moleong, J. 2014. Metodelogi Penelitian Kualitatif. Bandung: PT Remaja
Rosdakarya. Revell, L and James, A. (2007).Character Education in Schools and the Education
of Teachers.The Journal of Moral Education, Vol 36, No. 1. Subianto, J. 2013. Peran Keluarga, Sekolah, dan Masyarakat dalam Pembentukan
Karakter Berkualitas. Jurnal Penelitian Pendidikan Islam vol 8, No 2 Sugiyono. 2012. Metode Penelituan Kuantitatif, Kualitatif, dan R&D. Bandung:
Alfebet 193 Jurnal Multidisipliner Kapalamada Amanda 1(2), 187-194 Sutama. 2012. Metode Penelitian Pendekatan Kuantitatif, Kualitatif, PTK, R&D. Surakarta:Fairus Media
Suyadi. 2013. Strategi Pembelajaran Pendidikan Karakter. Bandung:PT Remaja
Rosdakarya
Zelvi, A. 2017. Proses Penanaman Nilai-nilai Agama Pada Anak Usia Dini dalam
Keluarga Di Kampung Gembira Pendeyan Umulharjo. Jurnal Pendidikan
anak Usia Dini, Vol 1, No 6. Sutama. 2012. Metode Penelitian Pendekatan Kuantitatif, Kualitatif, PTK, R&D. Surakarta:Fairus Media Zelvi, A. 2017. Proses Penanaman Nilai-nilai Agama Pada Anak Usia Dini dalam
Keluarga Di Kampung Gembira Pendeyan Umulharjo. Jurnal Pendidikan
anak Usia Dini, Vol 1, No 6. Zelvi, A. 2017. Sutama. 2012. Metode Penelitian Pendekatan Kuantitatif, Kualitatif, PTK, R&D.
Surakarta:Fairus Media
Suyadi. 2013. Strategi Pembelajaran Pendidikan Karakter. Bandung:PT Remaja
Rosdakarya
Zelvi, A. 2017. Proses Penanaman Nilai-nilai Agama Pada Anak Usia Dini dalam
Keluarga Di Kampung Gembira Pendeyan Umulharjo. Jurnal Pendidikan
anak Usia Dini, Vol 1, No 6. REKOMENDASI Proses Penanaman Nilai-nilai Agama Pada Anak Usia Dini dalam
Keluarga Di Kampung Gembira Pendeyan Umulharjo. Jurnal Pendidikan
anak Usia Dini, Vol 1, No 6. 194 194
|
https://openalex.org/W4387983538
|
https://dergipark.org.tr/en/download/article-file/3079955
|
English
| null |
Architectural Sustainability with cultural heritage values
|
Cultural heritage and science
| 2,023
|
cc-by-sa
| 4,052
|
Abstract Sustainability and cultural heritage are two interconnected and interdependent issues that
are essential for achieving sustainable development. Cultural heritage plays a critical role
in society as it is an integral part of the social and cultural fabric. This article focuses on the
relationship between sustainability and cultural heritage, specifically the integration of
cultural heritage into urban and environmentally sustainable development. To achieve
this, the study used Tabriz Bazaar as a case study to explore how cultural heritage can be
preserved while balancing economic expansion and sustainable development. The
research question is how cultural heritage can be integrated into urban and
environmentally sustainable development using Tabriz Bazaar as a case study. The
hypothesis is that the preservation and protection of cultural heritage values are crucial
for achieving sustainable development, and an interdisciplinary approach is necessary to
balance economic expansion with cultural heritage preservation. Using a case study
approach, the research collected data through a literature review and interviews with
experts in the fields of cultural heritage preservation, urban and environmental
sustainability, and economics. The research analyzes the challenges of balancing economic
development with cultural heritage preservation and proposes an interdisciplinary
approach to address these challenges. To make the area more sustainable and preserve its
cultural heritage values, specific measures such as promoting cycling in Tabriz Bazaar
were recommended. Moreover, education and awareness-raising programs are also
necessary to promote the importance of cultural heritage and its contribution to
sustainable development. Overall, the research highlights the interdependence between
sustainability and cultural heritage and emphasizes the importance of preserving cultural
heritage to achieve sustainable development. Policymakers, urban planners, and cultural
heritage conservationists can benefit from the findings of this study to develop strategies
that promote sustainable development while preserving cultural heritage values. Elyasi, S., & Yamaçlı, R. (2023). Architectural sustainability with cultural heritage values.
Cultural Heritage and Science, 4(2), 55-61 Cultural Heritage and Science
https://dergipark.org.tr/en/pub/cuhes
e-ISSN 2757-9050 Cultural Heritage and Science
https://dergipark.org.tr/en/pub/cuhes
e-ISSN 2757-9050 Sahar Elyasi *1
, Rusen Yamacli 1 1Eskişehir Technical University, Faculty of Architecture and Design, Department of Architecture, Eskişehir, Türki Keywords
Sustainability
Sustainable Development
Cultural Heritage
Cultural Heritage Management
Research Article
DOI: 10.58598/cuhes.1282179
Received:12.04.2023
Revised: 25.05.2023
Accepted:01.06.2023
Published:27.10.2023 Keywords
Sustainability
Sustainable Development
Cultural Heritage
Cultural Heritage Management
Research Article
DOI: 10.58598/cuhes.1282179
Received:12.04.2023
Revised: 25.05.2023
Accepted:01.06.2023
Published:27.10.2023 * Corresponding Author
*(elyasiiisahar@yahoo.com) ORCID ID 0000-0002-9982-2770
(yamaclir@gmail.com) ORCID ID 0000-0001-9659-9246 Cultural Heritage and Science – 2023, 4(2), 55-61 Cultural Heritage and Science – 2023, 4(2), 55-61 Architectural sustainability with cultural heritage values Sahar Elyasi *1
, Rusen Yamacli 1 1. Introduction tourism, increased real estate values, small business
improvement, and so on. Repurposing abandoned or
underutilized
historic
structures
is
critical
to
regenerating communities and enhancing quality of life. It is critical to acknowledge cultural heritage as a
significant resource and development incentive in order
to adopt sustainable development methods and improve
quality of life. The paper discusses the impact of
immovable cultural heritage on the implementation of
sustainable development strategies, the role of cultural
heritage in the context of globalization as a fundamental
means of avoiding the trend of cultural globalization, the
formation of sustainable communities, and the potential
impact of cultural heritage resources on economic
development and resource productivity. It is important Sustainable development encompasses not just
ecological but also economic, social, and cultural
dimensions. Many research has shown that preserving
cultural heritage enhances ecological, social, cultural, and
economic sustainability. Cultural legacy may benefit
communities' well-being and quality of life, assist in
alleviating the consequences of cultural globalization,
and serve as an incentive for long-term economic growth. Cultural heritage preservation is frequently viewed as a
barrier to economic development, despite the fact that
cultural heritage and its preservation can generate a
variety of economic benefits, including income and job
creation, professional training and the preservation of
craft skills, revitalization of city centers, heritage Cite this article (APA); Cultural Heritage and Science – 2023, 4(2), 55-61 economic and social roles as well, including as a tourist
attraction, source of employment, and source of revenue
[5]. It is also used to denote a cultural landscape that is
particular to a region or area. Cultural heritage and the
field of architectural science mutually constitute
interrelated components that exhibit a synergistic
relationship. A fresh outlook on cultural heritage has
materialized, evident in the consequential provisions of
the Faro Convention. Notably, the Convention's second
clause distinctly delineates cultural heritage as an
assemblage of historical resources, which individuals
perceive as an unattributed manifestation and mirror of
their continuously evolving principles, convictions,
erudition, and customs [6]. Cultural heritage is a dynamic
and evolving field, with new elements constantly being
added. Cultural heritage has a variety of roles and
functions that are related to the local community and to
the global community. Many of these roles are the same
as those of other sectors, such as tourism, arts, education,
and health [7]. However, these are sometimes
overlooked or undervalued. 2.1. Sustainable development Sustainable development aims to improve living
standards, preserve ecosystems, and secure basic needs
while creating a safer and happier future. It involves
designing economic, financial, trade, energy, agricultural,
and industrial policies that promote sustainability. This
includes investing in education, health, population, and
energy to avoid social debt for future generations. Sustainable development is a comprehensive concept
that impacts all aspects of human life, requiring
significant changes in national and international policies. Ultimately, it is an approach to economic growth that
seeks to meet the needs of the present without
compromising the ability of future generations to meet
their own needs [2]. The breadth of work in the name of sustainability, the
diversity of existing concepts, the importance of ordering
between principles, and the vast regional differences are
some of the first to compel the designer to find the truth. The common purpose of sustainability studies is to
examine from an architectural perspective how natural
energy resources can be protected and conserved by
assessing sustainability in relation to the natural
environment [3]. This includes reducing carbon
emissions, protecting natural resources, promoting
social equity, and ensuring economic growth that is both
environmentally responsible and socially inclusive. 1. Introduction They have a vital role to play
in society and are essential to maintaining cultural
identities and improving quality of life in the community
[8]. Key roles and functions of cultural heritage include
education
and
knowledge
creation,
economic
development, tourism, and aesthetic enjoyment. While
these are valuable contributions, they have not always
been considered in the same way that other sectors of
society have been treated. Some of the key challenges
facing the field of cultural heritage include globalization
and the loss of identity, changes in behavior and
attitudes. for cultural heritage cities and urban heritage cities to
recognize heritage preservation as a necessary goal for
sustainable planning in urban planning. Our past
experience shows to date that monument preservation
planning makes a positive contribution to sustainable
planning research. This is only possible by protecting and
enhancing key elements of the natural and cultural
heritage through planning [1]. Cultural heritage connects
us to the past, serves as the basis of our identity and is a
source of knowledge. 2.3. Cultural heritage and architectural sustainable
development Short-term
interests
promote
speculative
developments, which pose substantial dangers to
cultural assets and the historical landscape. Cultural
assets and historic surroundings are important not just
to individuals who own or reside in historic buildings. Cultural legacy may also contribute to a community's
well-being and quality of life by preventing cultural
globalization, preserving cultural variety, and positively
impacting economic growth. According to D. Rypkema, in
a larger perspective, the importance of heritage in the
quest of sustainable development is unmistakable: the
protection of cultural heritage offers environmental,
cultural, and economic sustainability [9]. Cultural
heritage plays a marginal role in the 2030 Agenda for
Sustainable Development. It is only explicitly mentioned
once in Goal 11, which relates to cities, specifically the
need to make cities and human settlements inclusive,
safe, resilient and sustainable through inclusive and
sustainable urbanization, planning and management
(Goal 11.3). and increased efforts to protect and preserve
the world's cultural and natural heritage (target 11.4). Sustainable development is a broad concept that
recognizes the interconnectedness of economic, social,
and environmental factors, and it has direct implications
for planning, housing, and urban development policies
worldwide. Urban planning is an effective tool for
steering sustainable development, but it alone is
insufficient to ensure sustainability. The United Nations'
Sustainable Development Goals (SDGs), particularly Goal
11, emphasize the need to create inclusive, safe, resilient,
and sustainable cities and communities. The New Urban
Agenda, adopted at the United Nations Conference on
Housing and Sustainable Urban Development, also
underscores the importance of participatory urban
planning and partnerships among stakeholders to
achieve sustainable development. 2.3.1.
Cultural heritage values in Tabriz The term cultural heritage is used to refers to the
“physical and intangible manifestations of a group’s
identity” [4]. This includes such elements as art,
architecture, stories, rituals, and so on. It has important According
to
the
trend
of
comprehensive
developments in today's cities of the world, the cities of
Iran and Azerbaijan in general and the city of Tabriz in 56 Cultural Heritage and Science – 2023, 4(2), 55-61 careful selection of suitable methodologies for the
project, and the seamless transmission of precise
information across different stages of scientific research. Additionally,
it
is
imperative
to
establish
a
comprehensive management and monitoring program
that
encompasses
the
pre-implementation,
implementation, and post-implementation phases of
conservation efforts [12]. In this regard, it is important to
draw up a plan for the protection of cultural heritage
values and then monitor its implementation to ensure
that it is carried out correctly. In Iran, there is not a
proper system for the management and protection of
cultural heritage. Thus, the contribution of this project is
to develop a comprehensive plan for the conservation of
cultural heritage in the country. In order to do this, it is
necessary to conduct a comprehensive assessment of
cultural values and identify their needs for protection
and preservation. This assessment will be based on the
available data and information. It will also be carried out
with the help of experts and authorities who have
relevant experience in the field. As a result, the project
will develop a model for the conservation and protection
of cultural heritage in Iran that can be used to ensure the
preservation of this valuable resource for future
generations. The work that is being done under this
project is expected to help Iran to make a valuable
contribution to the global community by ensuring the
conservation and preservation of its unique cultural
heritage. This will ultimately enable Iran to contribute to
the development of the global economy and enhance its
reputation as a developing country in the global
community. As a result, the success of the project will
help to enhance the country’s economy and improve the
overall quality of life for all citizens in Iran. particular have undergone many changes in the
architecture with historical value and the way of
distributing activities in recent decades. 2.3.1.
Cultural heritage values in Tabriz The role of the
city administration and planning in preserving historical
and unique architectures and creating a healthy
environment, economic, social, touristic city, etc. has
helped to make the urban system healthy and balanced. The historical city of Tabriz has experienced various
administrations and plans over the last few decades that
have left their impact on the development of the city and
the nature of the texture and environmental issues that
have caused the destruction of the old architecture and
the unequal distribution of the city services that these
Problems in different eras and time management were
different. Figure 1. The relationship between culture, society,
environment and economy with sustainability. Figure 1. The relationship between culture, society,
environment and economy with sustainability. Tabriz, located in northwestern Iran, has a rich
cultural heritage that reflects its historical importance as
a commercial center and cultural center. Some of the
main heritage values in Tabriz are shown in Table 1. The sustainability of historical bazaars is only
significant if it accurately reflects the everyday busyness
of the community through their physical structures. It is
crucial to consider how the renewal process affects the
daily life of the community. Unfortunately, urban
planners often make spatial changes in an effort to
enhance
functionality,
resulting
in
changes
or
elimination of current behavioral regulations. When the
workplaces of the sellers are significantly altered or
removed, the working methods, physical presence, and
sensory experiences they produce within the bazaar also
change, all of which are integral to the bazaar
environment and contribute to its individual memories
and collective identity [13]. Therefore, the sustainability
of historical bazaars can only be achieved if it accurately
portrays the hustle and bustle of daily life through their
physical structures, and if any renewal process considers
the impact on the community's daily life and preserves
the bazaar's unique character and identity. Cities are caught between the desire to be a member
of the global network and the need to maintain their
individuality and cultural origins. As new progress
symbols have shattered conventional contexts, new
meanings of community have developed [10]. The value of cultural heritage has always been an
important factor in the development of any country. In
fact, it has played an important role in the development
of nations. In addition, it has played an important role in
the development of the economies of these countries
[11]. 2.3.1.
Cultural heritage values in Tabriz In this sense, it has become a valuable resource for
both individual and social development. Accordingly, it is
important to develop a system for the protection and
management of these values. Therefore, developing a
system for the conservation of cultural heritage values is
a priority in Iran. To do this, it is necessary to identify
these values and then put measures in place to protect
them from harm. The successful execution of a
comprehensive
conservation
program,
involving
scientific surveys, analyses, and evaluations, entails the
diligent coordination of proficient technical experts, Rebuilding
Tabriz
market
with
sustainable
development in mind would involve considering
economic, social, and environmental aspects of the
project. Here are some steps that can be taken to achieve
this (Table 1). 57 Cultural Heritage and Science – 2023, 4(2), 55-61 Table 1. Heritage values of Tabriz. Items
Heritage values
y
z
1
Historic
Architecture
Tabriz has numerous historic
buildings, such as the Tabriz Historic
Bazaar Complex, the Blue Mosque and
the Arg of Tabriz, which reflect the
city's architectural and cultural
heritage. 2
Traditional
Crafts
Tabriz has a long history of producing
high-quality handicrafts such as
carpets, copperware and pottery,
which are highly valued both locally
and internationally. 3
Culinary
Traditions
Tabriz is known for its unique and
delicious cuisine, including dishes such
as Kofte Tabrizi, a type of meatball and
Ash Reshteh, a type of soup. Table 1. Heritage values of Tabriz. g
Items
Heritage values
y
z
1
Historic
Architecture
Tabriz has numerous historic
buildings, such as the Tabriz Historic
Bazaar Complex, the Blue Mosque and
the Arg of Tabriz, which reflect the
city's architectural and cultural
heritage. 2
Traditional
Crafts
Tabriz has a long history of producing
high-quality handicrafts such as
carpets, copperware and pottery,
which are highly valued both locally
and internationally. 3
Culinary
Traditions
Tabriz is known for its unique and
delicious cuisine, including dishes such
as Kofte Tabrizi, a type of meatball and
Ash Reshteh, a type of soup. 4
Festivals and
Celebrations
Tabriz hosts a number of annual
festivals and cultural celebrations
including the Tabriz International
Carpet Fair, the Tabriz Traditional
Music Festival and the Tabriz Pistachio
Festival. 5
Cultural
Diversity
Tabriz has a diverse population that
includes different ethnic and religious
groups that have contributed to the
city's cultural heritage over time. 3. Conclusion In
summary,
rebuilding
Tabriz
market
with
sustainable
development
in
mind
requires
a
comprehensive approach that considers economic,
social, and environmental factors. By implementing the
steps above, it's possible to create a sustainable market
that
promotes
economic
growth,
environmental
protection, and social well-being. 3. Educate the public: Educating the public about
the benefits of cycling and the availability of bike lanes in
the bazaar is essential to encourage more people to use
them. This can be done through outreach programs,
signage, and other forms of communication. Making the Tabriz bazaar more sustainable can have
a positive impact on cultural heritage in several ways. Here are some examples: 4. Provide bike parking facilities: Providing secure
and accessible bike parking facilities near the bazaar will
encourage people to cycle to the bazaar. This could
include bike racks, bike lockers, or even bike-sharing
programs. Preservation of historical architecture: The Tabriz
bazaar is a unique cultural heritage site with a rich
history
and
unique
architecture. By
promoting
sustainable practices in the bazaar, such as using
sustainable materials and reducing waste production,
the historical architecture of the bazaar can be preserved
and maintained for future generations. 5. Work with local businesses: Working with local
businesses to promote cycling to the bazaar can be a
powerful tool in encouraging people to use bike lanes. For example, businesses could offer discounts or
incentives to customers who arrive by bike. Promotion of local culture and heritage: Supporting
local businesses and promoting sustainable tourism
practices in the Tabriz bazaar can help to promote the
local culture and heritage of the region. This can include
showcasing traditional crafts and products, preserving
local customs and traditions, and celebrating local
festivals and events. 6. Collaborate
with
local
government:
Collaborating with local government to provide funding
and support for bike lanes and other cycling
infrastructure is critical to their success. This could
include providing funding for bike lane construction and
maintenance, as well as other initiatives to encourage
cycling, such as public awareness campaigns and bike
share programs. Increased awareness and education: By promoting
sustainable practices in the Tabriz bazaar, people can
become more aware of the importance of preserving
cultural heritage and the role that sustainability can play
in achieving this. This can lead to increased education
and awareness of cultural heritage among visitors and
residents of the bazaar. 2.3.1.
Cultural heritage values in Tabriz Items
Heritage values z Tabriz has numerous historic
buildings, such as the Tabriz Historic
Bazaar Complex, the Blue Mosque and
the Arg of Tabriz, which reflect the
city's architectural and cultural
heritage. 58 Cultural Heritage and Science – 2023, 4(2), 55-61
Figure 2. Iran‘s Map. Figure 3.Tabriz‘s Map. Figure 4. Tabriz Bazaar Map (cyling road). l
b l
l
h
f
d
h d
d Cultural Heritage and Science – 2023, 4(2), 55-61 Figure 3.Tabriz‘s Map. Figure 2. Iran‘s Map. Figure 3.Tabriz‘s Map. Figure 3.Tabriz‘s Map. Figure 2. Iran‘s Map. Figure 4. Tabriz Bazaar Map (cyling road). Figure 4. Tabriz Bazaar Map (cyling road). integrated into the design to reduce energy consumption
and promote clean energy use. Develop a sustainability plan: The first step is to
create a comprehensive plan that outlines the
sustainable goals and objectives of the rebuilding project. This plan should consider factors such as reducing
energy consumption, waste management, and promoting
sustainable economic growth. Promote local economy: To ensure the long-term
sustainability of the market, it's essential to promote the
local economy by supporting local businesses and
suppliers. This will create jobs and stimulate economic
growth in the region. Use sustainable materials: In the rebuilding process,
it's essential to use sustainable materials that have
minimal environmental impacts. For example, materials
that are locally sourced, recycled or have a low carbon
footprint. Encourage public transportation: The new design of
the market should encourage public transportation to
reduce the use of private cars, which contribute to air
pollution and traffic congestion. For example, bike lanes,
bus stops, or public transportation stations can be
incorporated into the design. Incorporate renewable energy sources: Renewable
energy sources like solar or wind power can be 59 Cultural Heritage and Science – 2023, 4(2), 55-61 Implement waste management strategies: The
market
should
have
a
comprehensive
waste
management
plan
that
includes
recycling
and
composting to minimize waste production and promote
sustainability. local culture and heritage, increasing awareness and
education, collaborating with local communities, and
promoting sustainable tourism practices. To encourage public transportation like bike lanes in
the bazaar of Tabriz, the following steps can be taken: 1. Conduct
a
feasibility
study:
Before
implementing any bike lanes or other infrastructure, it's
important to conduct a feasibility study to determine if
it's possible and practical to create bike lanes in the
bazaar. 2.3.1.
Cultural heritage values in Tabriz This study should consider factors like the size of
the bazaar, the number of visitors, and the availability of
alternative transportation options. Consider social aspects: In addition to economic and
environmental factors, social aspects such as inclusivity,
accessibility, and safety should be considered in the
rebuilding process. For example, the market should be
designed to be accessible to people with disabilities and
provide safe spaces for everyone. 2. Design safe and accessible bike lanes: The bike
lanes should be designed to ensure the safety of cyclists
and pedestrians, and to provide easy access to the bazaar. This includes separating the bike lane from vehicle
traffic, providing clear signage, and ensuring that the
bike lanes are well-maintained. 3. Conclusion By taking these steps, it's possible to encourage more
people to use public transportation like bike lanes in the
bazaar of Tabriz, reducing traffic congestion, improving
air quality, and promoting sustainable transportation
options. Collaboration with local communities: Making the
Tabriz bazaar more sustainable can involve working
closely with local communities to ensure that their
cultural heritage is respected and preserved. This can
involve
collaborating
with
local
artisans
and
craftspeople, supporting local festivals and events, and
providing opportunities for community engagement and
participation in sustainable practices. Author contributions Sahar Elyasi: Methodology, Field study, writing original
draft preparation Ruşen Yamaçlı: Writing-Reviewing
and Editing, validation Sustainable tourism: Sustainable tourism practices
can help to promote cultural heritage in the Tabriz
bazaar, such as supporting local businesses, providing
opportunities for cultural exchange and learning, and
minimizing negative impacts on the environment and
local communities. Conflicts of interest There is no conflict of interest between the authors. In summary, making the Tabriz bazaar more
sustainable can have a positive impact on cultural
heritage by preserving historical architecture, promoting 60 Cultural Heritage and Science – 2023, 4(2), 55-61 References 7. Sułkowski, Ł. (2020). Covid-19 Pandemic; Recession,
Virtual Revolution Leading to De-globalization? Journal of Intercultural Management, 12(1), 1-11. 1. Nocca, F. (2017). The role of cultural heritage in
sustainable
development:
Multidimensional
indicators as decision-making tool. Sustainability,
9(10), 1882. 8. Streimikiene, D., Mikalauskiene, A., & Kiausiene, I. (2019). The impact of value created by culture on
approaching the sustainable development goals: Case
of the Baltic States. Sustainability, 11(22), 6437. 2. Elyasi, S., & Yamacli, R. (2022). Investigation of
Iranian
residential
buildings
with
sustainable
architecture in hot and dry areas. Journal of
Architecture, Engineering & Fine Arts, 4(1), 16-29. 9. Rypkema,
D. (2005). Cultural
heritage
and
sustainable economic and social development. Europa Nostra. 10. Banerjee, T., & Southworth, M. (1990). City Sense and
City Design. The MIT Press. 3. Çiğan, A., & Yamaçlı, R. (2020). Doğal Enerji,
Sürdürülebilir Kalkınma ve Mimarlık Politikaları. Duzce University Journal of Science and Technology,
8, 554_571 11. Seyhan, A. K., & Badem, A. (2018). Erzincan ilindeki
hayvansal
atıkların
biyogaz
potansiyelinin
araştırılması. Academic
Platform-Journal
of
Engineering and Science, 6(1), 25-35. 4. Bonenberg, W. (2019). The Role of Cultural Heritage
in Sustainable Development. Values and Valuation as
Key Factors in Spatial Planning of Rural Areas. Advances in Human Factors in Architecture,
Sustainable Urban Planning and Infrastructure (s. 124-134). Advances in Intelligent Systems and
Computing book series. 12. Naycı, N., Abruzzese, D., Güler, C., Tağa, H.,
Cammarano, P., Vuth, S., & Türkoğlu, H. (2020). Multidisciplinary Researches in Cultural Heritage
Studies: An Approach on Akkale Cistern in Erdemli,
Mersin. Cultural Heritage and Science, 1(1), 15-22. 13. Karataş, L. (2022). Conservation status of intangible
cultural heritage after restoration: Case study of
Mardin
Spice
Bazaar. Cultural
Heritage
and
Science, 3(2), 30-36. 5. Ashfaq, J., Hassan, H., & Khan, A. (2022). The Impact of
Mass Media, Word-Of-Mouth on Travel Intention and
Mediating Role of Destination Image and Tourist
Attitude. International Journal of Academic Research
in Business and Social Sciences, 3224-3239. 6. Tepebaş,
U. (2021). The
Contribution
of
Archaeological Surveys on the Perception of Cultural
Heritage: Cilicia as a Case Study. Cultural Heritage and
Science, 25-30. © Author(s) 2023. This work is distributed under https://creativecommons.org/licenses/by-sa/4.0/ 61
|
https://openalex.org/W1784904721
|
https://europepmc.org/articles/pmc1481587?pdf=render
|
English
| null |
Role of transcription factor Sp1 and CpG methylation on the regulation of the human podocalyxin gene promoter
|
BMC molecular biology
| 2,006
|
cc-by
| 9,197
|
BioMed Central BioMed Central BioMed Central Open Ac
Research article
Role of transcription factor Sp1 and CpG methylation on the
regulation of the human podocalyxin gene promoter
Nora Butta, Susana Larrucea, Sonia Alonso, Ramón B Rodriguez,
Elena G Arias-Salgado, Matilde S Ayuso, Consuelo González-Manchón and
Roberto Parrilla* Address: Department of Physiopathology and Human Molecular Genetics Centro de Investigaciones Biologicas (CSIC), Ramiro de Maeztu 9
28040-Madrid, Spain Email: Nora Butta - nbutta@cib.csic.es; Susana Larrucea - slarrucea@cib.csic.es; Sonia Alonso - soniaalm@cib.csic.es;
Ramón B Rodriguez - rbrod@cib.csic.es; Elena G Arias-Salgado - egas@cib.csic.es; Matilde S Ayuso - msayuso@cib.csic.es; Consuelo González-
Manchón - cgmanchon@cib.csic.es; Roberto Parrilla* - rparrilla@cib.csic.es * Corresponding author Published: 09 May 2006
BMC Molecular Biology 2006, 7:17
doi:10.1186/1471-2199-7-17
Received: 08 February 2006
Accepted: 09 May 2006
This article is available from: http://www.biomedcentral.com/1471-2199/7/17
© 2006 Butta et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0),
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Received: 08 February 2006
Accepted: 09 May 2006 BMC Molecular Biology 2006, 7:17
doi:10.1186/1471-2199-7-17
This article is available from: http://www.biomedcentral.com/1471-2199/7/17 This article is available from: http://www.biomedcentral.com/1471-2199/7/17 This article is available from: http://www.biomedcent © 2006 Butta et al; licensee BioMed Central Ltd. © 2006 Butta et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0),
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. © 006
utta et a ; ce see
o e Ce t a
t . This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creative
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cit BMC Molecular Biology Open Access Background g
Podocalyxin (podxl) is an integral membrane protein
originally identified by Kerjaschki et al [1] in the foot
processes of the rat kidney glomerular podocytes. It was
described as a sialoprotein of 140-kDa based on its stain-
ing with alcian blue. The high sulfate content of this pro-
tein contributes in part to its heavily negative charge [2]
that is supposed to maintain open the interdigitations of
the podocyte foot for the urine filtration. The importance
of the negative charge of this protein is indicated by the
loss of the structure and function of the podocytes by
treatment with polycations or desialylation with neurami-
nidase [3,4]. Null mice for podxl die soon after birth due
to failure to develop foot processes in the glomerular epi-
thelial cells [5]. Podxl like proteins have been cloned from
the rabbit, human, chicken and mouse [6-9]. The podxl proteins show a poorly conserved aminotermi-
nal, extracellular, mucin-like domain containing multiple
N- and O-linked glycosylation sites, a disulfide containing
domain and conserved transmembrane and cytoplasmic
domains. The cytoplasmic tails showed the highest iden-
tities among these proteins and interaction with cytosolic
proteins have been reported. Based on structural similari-
ties and their high content in sialic acid, Sassetti et al [13]
proposed that CD34, podxl and endoglycan, would form
a family named the CD34 family of sialomucins. The
three proteins are found in the vascular endothelia and
also in early hematopoietic precursors, suggesting that
their function may go from either cell anti-adherence, as it
is the case in the renal podocytes, to selective adherence or
control of cellular differentiation. The podxl is also expressed in the luminal face of the vas-
cular endothelia, hematopoietic cells and platelets
[10,11], among other tissues. The function of this protein
in non-renal cells has not yet been elucidated. It has been
suggested that it may function as a ligand for leucocyte
adhesion molecule L-selectin [12] due to its location in
the postcapillary venules of the lymph node, high
endothelial venules, and the fact that most of the L-selec-
tin ligands in the vascular endothelium are sialomucins. Despite the interest of podxl no much is known about the
transcriptional regulation of its gene in different cells. Abstract Background: Podocalyxin (podxl) is a heavily glycosylated transmembrane protein mainly found
on the apical membrane of rat podocytes and also in endothelial, hematopoietic, and tumor cells. Despite of its interest no much is known about the transcriptional regulation of podxl in different
cells. Thus, we aimed at studying the functional features of the 5'-regulatory region of the human
Podxl gene. Results: The promoter region of the human Podxl gene has been cloned and its structure and
function were analyzed. The primary DNA sequence is rich in G+C and is devoid of TATA or
CAAT boxes. The sequence contains recognition sites for several putative transcription factors;
however, the basic promoter activity seems to rely entirely on Sp1 transcription factor since
supershift analysis was positive only for this factor. The region encompassed by 66 to -111 nts
conferred the minimal transcriptional activity that increases as the number of Sp1 sites augmented
with the length of the promoter fragment. In Sp1-lacking insect cells the Podxl promoter constructs
showed activity only if cotransfected with an Sp1 expression plasmid. Finally, mutation of the Sp1
sites reduced the promoter activity. We analyzed whether methylation of the CpG dinucleotides present in the first ~600 nts of the
promoter region of Podxl could explain the variable rates of expression in different types of cells. Inactivation of methyltransferases by 5'-aza-2'deoxicitidine showed a dose-dependent increase in
the podxl content. Moreover, in vitro methylation of the promoter constructs -111,-181 and -210
led to an almost complete reduction of the promoter activity. A correlation was found between
the degree of methylation of the CpG promoter dinucleotides and the rate of podxl expression in
different cell lines. Conclusion: Our results indicate that transcriptional regulation of Podxl is supported primarily by
Sp1 site(s) and that DNA-methylation of the CpG promoter islands contributes to control the
tissue specific expression of podxl. Page 1 of 12
(page number not for citation purposes) (page number not for citation purposes) BMC Molecular Biology 2006, 7:17 http://www.biomedcentral.com/1471-2199/7/17 Thus, the podxl of the vascular endothelia may contribute
to the migration of lymphocytes toward the lymph nodes. Background The
binding of the Wilms Tumor suppressor factor (WT1) to
conserved elements within the Podxl gene promoter
results in a potent transcriptional activation [14] that
seems to be relevant only for the induction and develop-
ment of the kidney; however, a detailed analysis of the
human Podxl promoter (Podxl-pr) has not yet been per- Transient transfection of deletion mutants of the 5'-flanking region of the human podocalyxin gene
Figure 1
Transient transfection of deletion mutants of the 5'-flanking region of the human podocalyxin gene. Description
of the construction of the pGL3-Podxl-pr reporter plasmids and transient transfection experiments are described under
"Methods". Luciferase activity was corrected for transfection efficiency with the values obtained with Renilla and was expressed
as percent of the maximal activity of the -364 construct. The results are mean ± SEM of three experiments performed in dupli-
cates. g
g
p
y
g
g
Transient transfection of deletion mutants of the 5'-flanking region of the human podocalyxin gene. Description
of the construction of the pGL3-Podxl-pr reporter plasmids and transient transfection experiments are described under
"Methods". Luciferase activity was corrected for transfection efficiency with the values obtained with Renilla and was expressed
as percent of the maximal activity of the -364 construct. The results are mean ± SEM of three experiments performed in dupli-
cates. Page 2 of 12
(page number not for citation purposes) http://www.biomedcentral.com/1471-2199/7/17 http://www.biomedcentral.com/1471-2199/7/17 BMC Molecular Biology 2006, 7:17 Western blots of human podocalyxin and Sp1 transcription
factor
Figure 2
Western blots of human podocalyxin and Sp1 tran-
scription factor. Cell lysates (20 µg) were resolved on
SDS-7.5% polyacrylamide gels under reducing conditions,
transferred to PVDF membranes and immunoblotted with
either 1 µg/ml of anti-podxl or anti Sp1 antibodies. β-Actin
content was blotted as a control of gel loading. The experi-
ment was repeated three times. region and a lack of CCAAT and TATA-like elements, a fea-
ture commonly found in the house-keeping genes. We
also detected potential binding sites for the transcription
factors AP2 (-247/-236 and -114/-93 nts) (two overlapped
sites); overlapped Sp1 sites (167/-140, -146/-128 and -91/
-68 nts); overlapped Sp1 and AP2 sites (-279/-266 nts),
overlapped Sp1 and NFKB sites (-204/-189 nts), three
GATA-1 sites (-1138/-1129, -1057/-1051 and -444/-435
nts) and three WT-1 sites (two overlapped at -1233/-1216
and one at -507/-498 nts). Regulatory role of Sp1 on Podxl promoter activity Regulatory role of Sp1 on Podxl promoter activity
The fragments up to the first 364 bp show a progressive
increasing number of Sp1 sites, suggesting their role in
transcription. To test this possibility we transfected some
of the constructs into the Sp1-deficient Drosophila-
derived Schneider's SL2 cells. As indicated in Fig. 3, no
transcriptional activity could be detected when the con-
structs +66, -111, -181,-210 and -364 were transfected
into these cells; however, the transcriptional activity was
initiated in all the constructs, except for the +66, when the
cells were cotransfected with a plasmid expressing Sp1
(Fig. 3). Thus, the Sp1sites seem to be essential to main-
tain a normal basal rate of transcription of Podxl. formed. In the present study we have further characterized
the 5' regulatory region of this gene. We found absence of
a TATA-box and a transcriptional regulation supported
primarily by Sp1 site(s). In addition, the role of cytosine
methylation of the promoter region has been analyzed to
elucidate whether this epigenetic mechanism might con-
tribute to control the specific regulation of the expression
of this gene. formed. In the present study we have further characterized
the 5' regulatory region of this gene. We found absence of
a TATA-box and a transcriptional regulation supported
primarily by Sp1 site(s). In addition, the role of cytosine
methylation of the promoter region has been analyzed to
elucidate whether this epigenetic mechanism might con-
tribute to control the specific regulation of the expression
of this gene. Background To determine the location of the promoter activity we
studied the transcriptional activity of chimeric constructs
of progressively 5'-deleted DNA fragment (-1297/231)
linked to the luciferase (pGL3) reporter (Fig. 1). The con-
structs were transiently transfected into Tera-1, HEK293
and HepG2 cells, which podxl content differs considera-
bly (Fig. 2). Figure 1 depicts the results obtained with the
three types of cells. No significant transcriptional activity
was detected with the fragment of +66 bp. The activity
started on construct -111, increasing with the length of the
fragment up to -364. Thus, the basal promoter activity is
located within the -111 to +66 bp region, whereas the
upstream 253 bp confers maximal transcriptional activity. The extension of the 5' flanking sequence up to -1297
diminished the promoter activity. All cells expressed sim-
ilar amounts of Sp1 (Fig. 2, middle row). Western blots of human podocalyxin and Sp1 transcription
factor
Figure 2
Western blots of human podocalyxin and Sp1 tran-
scription factor. Cell lysates (20 µg) were resolved on
SDS-7.5% polyacrylamide gels under reducing conditions,
transferred to PVDF membranes and immunoblotted with
either 1 µg/ml of anti-podxl or anti Sp1 antibodies. β-Actin
content was blotted as a control of gel loading. The experi-
ment was repeated three times. p
y
p
p
g
Western blots of human podocalyxin and Sp1 tran-
scription factor. Cell lysates (20 µg) were resolved on
SDS-7.5% polyacrylamide gels under reducing conditions,
transferred to PVDF membranes and immunoblotted with
either 1 µg/ml of anti-podxl or anti Sp1 antibodies. β-Actin
content was blotted as a control of gel loading. The experi-
ment was repeated three times. Features of the 5' flanking region of the human
podocalyxin The DNA/protein interactions were characterized by elec-
trophoretic mobility of DNA-protein complexes (EMSA). The assays were performed with two different probes
encompassing the -243/-18 and -538/-238 Podxl frag-
ments and nuclear extracts from cells with either low or
high levels of podxl expression (Meg0l and Tera-1 cells,
respectively, Fig. 4A). Nuclear extracts from Meg0l and
Tera-1 cells formed retardation complexes with the -243/-
18 and -538/-238 probes. The identity of proteins inter-
acting at the potential binding sites was assessed using
competitor oligonucleotides for AP2, Sp1, GATA-1 and
NFκB sites and a non-specific unrelated oligonucleotide. AP2 as well as Sp1 oligonucleotides prevented the forma- The human Podxl gene and 250 bps upstream of the puta-
tive start methionine was previously cloned and mapped
to chromosome 7q32-q33 by Kershaw et al [7,15]. We
amplified by PCR a 1528 bp-DNA fragment of the 5' reg-
ulatory region of the human Podxl gene comprising 1297
bp from the transcription start site plus 231 bp of 5'-
untranslated region, using the primers sense: -1297/-1275
and antisense: 231/208. We started numbering from the
transcription start site [GenBank:AF395890]. Computer
analysis using the software "Transcription Factor Binding
Site Profile Database" (TFSEARCH program) [16],
revealed a high G+C content of the proximal promoter Page 3 of 12
(page number not for citation purposes) BMC Molecular Biology 2006, 7:17 http://www.biomedcentral.com/1471-2199/7/17 Effect of Sp1 on podocalyxin gene promoter activity in insect
cells
Figure 3
Effect of Sp1 on podocalyxin gene promoter activity
in insect cells. Drosophila-derived SL2 cells were transiently
cotransfected with 5 µg of pGL3/Podxl-pr constructs and
with either 0.5 µg of pPAC void plasmid or 0.5 µg of pPAC-
Sp1 plasmid. Luciferase activities were normalized by protein
concentration and are shown as the fold increases relative to
the basal activity of void pGL3-basic plasmid (control). Values
reported in the figure are mean ± SEM of three independent
experiments in duplicates. Gel mobility-shift analysis of DNA-protein complexes using
the human podocalyxin promoter fragment -243/-18
Figure 4
Gel mobility-shift analysis of DNA-protein complexes
using the human podocalyxin promoter fragment -
243/-18. A) Western blots were performed as described
under Fig. 2; B) Five µg of nuclear extracts (NE) from Tera-1
and Meg0l cells were incubated with the probe. Competitive
gel shift assay was done in the presence of 100-fold molar
excess of unlabeled double-stranded oligonucleotides corre-
sponding to the pointed out transcription factors consensus
and unrelated nonspecific (Non Sp) motifs. Effect of Sp
cells
Figure 3 Competitive
gel shift assay was done in the presence of 100-fold molar
excess of unlabeled double-stranded oligonucleotides corre-
sponding to the pointed out transcription factors consensus
and unrelated nonspecific (Non Sp) motifs. Antibodies (2 µg)
for AP2 or non-specific were preincubated with the Tera-1
and Meg0l NE before the addition of the Podxl probe. Gel mobility-shift analysis of DNA-protein complexes using
the human podocalyxin promoter fragment -243/-18
Figure 4
Gel mobility-shift analysis of DNA-protein complexes
using the human podocalyxin promoter fragment -
243/-18. A) Western blots were performed as described
under Fig. 2; B) Five µg of nuclear extracts (NE) from Tera-1
and Meg0l cells were incubated with the probe. Competitive
gel shift assay was done in the presence of 100-fold molar
excess of unlabeled double-stranded oligonucleotides corre-
sponding to the pointed out transcription factors consensus
and unrelated nonspecific (Non Sp) motifs. Antibodies (2 µg)
for AP2 or non-specific were preincubated with the Tera-1
and Meg0l NE before the addition of the Podxl probe. We ran supershift assays using nuclear extracts from
Meg0l cells on different probes: "A" probe includes -91/-
68 overlapped Sp1 sites (Sp1-A); "B" probe includes -146/
-128 Sp1 sites (Sp1-B); "C" probe includes -146/-128 and
-167/-140 sites (Sp1-B and Sp1-C), and "D" probe that
only includes -204/-189 Sp1 site (Sp1-D, Fig. 5A). As
shown in Fig. 5B, protein-DNA complexes were formed
with all the probes tested and at least one of the com-
plexes was shifted by anti-Sp1, excepting those in which
probe B were used. Therefore, the binding of nuclear pro-
teins to the Sp1-C site may explain the supershifted com-
plexes observed with "C" probe. When probes with
mutated Sp1 sites were used no supershifts were observed
(Fig. 5B). Similar results were obtained when Tera-1
nuclear extracts were used (data not shown). In agreement
with the EMSA assays, mutations in either Sp1-A, Sp1-C or
Sp1-D sites inhibited the activity of the Podxl promoter by
~50% (Fig. 6) whereas mutation of Sp1-B did not (data
not shown). Effect of Sp
cells
Figure 3 Effect of Sp
cells
Figure 3 p
p
y
g
p
y
g
Effect of Sp1 on podocalyxin gene promoter activity
in insect cells. Drosophila-derived SL2 cells were transiently
cotransfected with 5 µg of pGL3/Podxl-pr constructs and
with either 0.5 µg of pPAC void plasmid or 0.5 µg of pPAC-
Sp1 plasmid. Luciferase activities were normalized by protein
concentration and are shown as the fold increases relative to
the basal activity of void pGL3-basic plasmid (control). Values
reported in the figure are mean ± SEM of three independent
experiments in duplicates. p
p
y
g
p
y
g
Effect of Sp1 on podocalyxin gene promoter activity
in insect cells. Drosophila-derived SL2 cells were transiently
cotransfected with 5 µg of pGL3/Podxl-pr constructs and
with either 0.5 µg of pPAC void plasmid or 0.5 µg of pPAC-
Sp1 plasmid. Luciferase activities were normalized by protein
concentration and are shown as the fold increases relative to
the basal activity of void pGL3-basic plasmid (control). Values
reported in the figure are mean ± SEM of three independent
experiments in duplicates. tion of the same protein/DNA complex within the -243/-
18 probe (Fig. 4B), whereas no effect was observed (data
not shown) with any of the other oligonucleotides. No
displacement of the EMSA bands was appreciated with an
antibody against AP2, regardless of the type of nuclear
extract used (Fig. 4B). With the -538/-238 probe no com-
petition was observed with a 100-fold excess of oligonu-
cleotides (data not shown). tion of the same protein/DNA complex within the -243/-
18 probe (Fig. 4B), whereas no effect was observed (data
not shown) with any of the other oligonucleotides. No
displacement of the EMSA bands was appreciated with an
antibody against AP2, regardless of the type of nuclear
extract used (Fig. 4B). With the -538/-238 probe no com-
petition was observed with a 100-fold excess of oligonu-
cleotides (data not shown). Gel mobility-shift analysis of DNA-protein complexes using
the human podocalyxin promoter fragment -243/-18
Figure 4
Gel mobility-shift analysis of DNA-protein complexes
using the human podocalyxin promoter fragment -
243/-18. A) Western blots were performed as described
under Fig. 2; B) Five µg of nuclear extracts (NE) from Tera-1
and Meg0l cells were incubated with the probe. Features of the 5' flanking region of the human
podocalyxin Antibodies (2 µg)
for AP2 or non-specific were preincubated with the Tera-1
and Meg0l NE before the addition of the Podxl probe. tion of the same protein/DNA complex within the -243/-
18 probe (Fig. 4B), whereas no effect was observed (data
not shown) with any of the other oligonucleotides. No
displacement of the EMSA bands was appreciated with an
antibody against AP2, regardless of the type of nuclear
extract used (Fig. 4B). With the -538/-238 probe no com-
petition was observed with a 100-fold excess of oligonu-
cleotides (data not shown). We ran supershift assays using nuclear extracts from
Meg0l cells on different probes: "A" probe includes -91/-
68 overlapped Sp1 sites (Sp1-A); "B" probe includes -146/
(
)
b
l d
/
d
Gel mobility-shift analysis of DNA-protein complexes using
the human podocalyxin promoter fragment -243/-18
Figure 4
Gel mobility-shift analysis of DNA-protein complexes
using the human podocalyxin promoter fragment -
243/-18. A) Western blots were performed as described
under Fig. 2; B) Five µg of nuclear extracts (NE) from Tera-1
and Meg0l cells were incubated with the probe. Competitive
gel shift assay was done in the presence of 100-fold molar
excess of unlabeled double-stranded oligonucleotides corre-
sponding to the pointed out transcription factors consensus
and unrelated nonspecific (Non Sp) motifs. Antibodies (2 µg)
for AP2 or non-specific were preincubated with the Tera-1
and Meg0l NE before the addition of the Podxl probe. Effect of Sp1 on podocalyxin gene promoter activity in insect
cells
Figure 3
Effect of Sp1 on podocalyxin gene promoter activity
in insect cells. Drosophila-derived SL2 cells were transiently
cotransfected with 5 µg of pGL3/Podxl-pr constructs and
with either 0.5 µg of pPAC void plasmid or 0.5 µg of pPAC-
Sp1 plasmid. Luciferase activities were normalized by protein
concentration and are shown as the fold increases relative to
the basal activity of void pGL3-basic plasmid (control). Values
reported in the figure are mean ± SEM of three independent
experiments in duplicates. Effect of promoter CpG methylation on the cellular
expression of podxl Bisulfite changes all the unmethylated cytosines to uracils
and the uracils are PCR-amplified as thymidines, whereas
the methylated cytosines are not modified. As indicated in
Fig. 7, Tera-1 and Meg0l cells showed different patterns of
methylation. In high-podxl expressing cells the promoter
region is unmethylated whereas in low-podxl expressing
cells, Meg0l cells, the promoter is partially methylated. The main difference was observed in the methylation of
CpG boxes from 23 to 27 (Fig. 7). (Meg0l) levels of podxl (Fig. 4A). Genomic DNA of both
cell lines were treated with bisulfite and then amplified by
PCR with the specific primers described under Methods. Bisulfite changes all the unmethylated cytosines to uracils
and the uracils are PCR-amplified as thymidines, whereas
the methylated cytosines are not modified. As indicated in
Fig. 7, Tera-1 and Meg0l cells showed different patterns of
methylation. In high-podxl expressing cells the promoter
region is unmethylated whereas in low-podxl expressing
cells, Meg0l cells, the promoter is partially methylated. The main difference was observed in the methylation of
CpG boxes from 23 to 27 (Fig. 7). To further assess the influence of CpG methylation on the
promoter activity of Podxl, we methylated promoter frag-
ments in vitro by Sss I followed by their transfection into
HEK293 cells. Fig. 9 shows the results of these experi-
ments using either mock methylated or methylated -111,-
181 and -210 constructs. The in vitro methylation of either
construct by Sss I almost abolished the promoter activity. Effect of promoter CpG methylation on the cellular
expression of podxl DNA methylation has been suggested to play a role in
maintaining the stability of cell differentiation and in reg-
ulating gene transcription [17,18]. It has been recently
reported that the selective demethylation of a promoter
DNA may be a general mechanism required for the repro-
gramming of somatic cell nuclei [19]. DNA sequences rich
in CpG are susceptible of methylation and therefore
silencing the gene expression. Computer analysis of the
5'-flanking region of the Podxl gene with CpGPlot pro-
gram [20] shows the existence of a putative CpG island,
from -607 to the translation start site, containing 27 CpG
dinucleotide sequences. The methylation of cytosines
may control the transcriptional activity through changes
in the binding activity of Sp1 sites. Thus, in order to eluci-
date the possible epigenetic regulation of Podxl we ana-
lyzed the methylation status of individual promoter CpG
sites in cells expressing either high (Tera-1) or low Page 4 of 12
(page number not for citation purposes) Page 4 of 12
(page number not for citation purposes) BMC Molecular Biology 2006, 7:17 http://www.biomedcentral.com/1471-2199/7/17 Identification of Sp1-DNA complexes by supershift assays
Figure 5
Identification of Sp1-DNA complexes by supershift assays. A) Different probes were designed as described under
"Methods" in order to identify functional Sp1 sites in the Podxl promoter. B) Antibodies (2 µg) for Sp1 and non-specific, were
preincubated with 5 µg of Meg01 nuclear extracts (NE) before the addition of the wild or mutated Sp1-containing Podxl
probes. BSA (2 µg) was used as a control. An arrow points to the supershifted Sp1 bands. e t cat o o Sp
co p e es by supe s
t assays
gu e 5
Identification of Sp1-DNA complexes by supershift assays. A) Different probes were designed as described under
"Methods" in order to identify functional Sp1 sites in the Podxl promoter. B) Antibodies (2 µg) for Sp1 and non-specific, were
preincubated with 5 µg of Meg01 nuclear extracts (NE) before the addition of the wild or mutated Sp1-containing Podxl
probes. BSA (2 µg) was used as a control. An arrow points to the supershifted Sp1 bands. was observed in Meg0l cells. Treatment with 5-azaC did
not modify the Sp1 cellular content (Fig. 8B). (Meg0l) levels of podxl (Fig. 4A). Genomic DNA of both
cell lines were treated with bisulfite and then amplified by
PCR with the specific primers described under Methods. Discussion In this work we have analyzed the structure and function
of the Podxl gene promoter. This protein was originally
described in the podocytes of the epithelial glomerular
cells [1]. Podxl is also present in extra-renal tissues, partic-
ularly in the vascular endothelial and hematopoietic cells. Moreover, podxl has also been found in tumor cells [21-
23] and its expression has been considered as a good The role of methylation in controlling the Podxl gene was
also investigated studying the effects of a demethylating
agent, 5'-aza-2' deoxycitidine (5-azaC), on the podxl
expression. Fig. 8A shows that 5-azaC was unable to
enhance the expression of podxl in HEK293 cells. In con-
trast, a 5-azaC-concentration-dependent increase of podxl Page 5 of 12
(page number not for citation purposes) Page 5 of 12
(page number not for citation purposes) BMC Molecular Biology 2006, 7:17 http://www.biomedcentral.com/1471-2199/7/17 Effect of mutating the Sp1 recognition sites on the activity of podocalyxin promoter
Figure 6
Effect of mutating the Sp1 recognition sites on the activity of podocalyxin promoter. Transient transfections were
performed into HEK293 cells. Results represent mean ± SEM from four independent experiments performed in duplicates. Promoter activity of each transfection was expressed as a ratio of luciferase/renilla activities. Data are expressed as percent of
the maximal activity of the -210 construct *p < 0.05 vs. non Sp1-site-mutated construct. Effect of mutating the Sp1 recognition sites on the activity of podocalyxin promoter
Figure 6
Effect of mutating the Sp1 recognition sites on the activity of podocalyxin promoter. Transient transfections were
performed into HEK293 cells. Results represent mean ± SEM from four independent experiments performed in duplicates. Promoter activity of each transfection was expressed as a ratio of luciferase/renilla activities. Data are expressed as percent of
the maximal activity of the -210 construct *p < 0.05 vs. non Sp1-site-mutated construct. (Fig. 1). This observation may find an explanation in the
similar levels of Sp1 factor found in these cells (Fig. 2). marker of mieloproliferative disorders [24]. The interest
on podxl has been recently focused on its putative capac-
ity to regulate the intercellular contacts. The nucleotide competition assays gave positive results
for Sp1 and AP2 whether nuclear extracts from Tera-1 or
from Meg0l cells were used; however, no supershift was
observed when monoclonal against AP2 was utilized (Fig. 4B). Discussion The competitive effect of AP2 oligonucleotide in the
formation of protein-DNA complexes may be related to
the sequence similarities between Sp1 and AP2 recogni-
tion sites. The EMSA studies indicate that overlapped Sp1
sites -167/-140 nts, -91/-68 nts and -204/-189 nts were
able to interact with Sp1, since some of the formed com-
plexes were supershifted by anti-Sp1 Ab and their forma-
tion was-prevented by mutating the recognition sites. Moreover, the mutation of the Sp1 sites reduced the tran-
scriptional activity of Podxl. The information about the regulation of the Podxl gene is
scarce. The Wilms tumor suppressor factor (WT1) is a
potent regulator of kidney induction and nephrogenesis
[25,26] that controls the expression of Podxl [14]. How-
ever, the WT1 factor is downregulated in all tissues except
in the glomerular epithelial cells where it is expressed
throughout life. Mutations in the WT1 gene are associated
with glomerulosclerosis in patients of Denys Drash syn-
drome indicating the importance of this factor in the renal
function in adulthood. Moreover, nephrogenesis can be
rescued in WTl-null mice by reintroducing the gene [26]. However, regulatory factors other than WT1 in renal or
extra-renal tissues are undefined. The lack of TATA or
CAAT boxes in the Podxl promoter, its high content of
C+Gs and the control by Sp1 sites are typical features of
house-keeping genes. The transcriptional activity of the
progressive 5' deletions of this promoter indicated that its
basic activity was located in the region 66 to -111 nts
increasing as the length of the DNA fragment increased
(Fig. 1). The progressive rise in the promoter activity
directly correlates with the number of recognition sites for
Sp1, suggesting its functional importance. This interpreta-
tion is supported by the absence of transcriptional activity
of the Podxl promoter constructs in insect cells (Dro-
sophila SL2) that lack Sp1 transcription factor and the ini-
tiation of transcriptional activity by cotransfection of an
expression plasmid encoding Sp1 (Fig. 3). Despite differ-
ences in endogenous content of podxl, the activity of the
Podxl promoter was similar whether Tera-1, HepG2 or
HEK293 cells were used in the transfection experiments DNA methylation is associated with malignant transfor-
mation [27] of cells but knowledge of the epigenetic
events in the initiation and progression of human cancer
is limited. The abundant number of CpG islands in the
Podxl promoter suggested that methylation could be a
regulatory factor. Conclusion The in vitro transcription of the human Podxl promoter is
dependent on the presence of Sp1 sites. This conclusion is
supported by the following observations: 1) No activity
was detected when the promoter constructs were trans-
fected into Sp1-lacking insect cells in which the transcrip-
tion was initiated by cotransfection of an Sp1 expression
plasmid; 2) The promoter function was impaired by
mutation of the Sp1 sites. Methylation of the CpG promoter islands seems to be a
significant regulatory mechanism in controlling the tran-
scription of Podxl. The latter observation may provide an
explanation for the different rates of podxl expression in
different tissues of the organism. Expression
treated wi
Figure 8 Expression of human podocalyxin in HEK293 and
Meg0l cells treated with 5-AzaC. Cells were treated
with 5-AzaC for 48 hrs. Cell lysates were resolved on SDS-
7.5% polyacrylamide gels under non-reducing conditions,
transferred to PVDF membranes and immunoblotted with A)
1 µg/ml of anti-podocalyxin mAb and B) 1 µg/ml of anti-Sp1
mAb. β-Actin was blotted as a loading control. The experi-
ment was repeated three times. http://www.biomedcentral.com/1471-2199/7/17 BMC Molecular Biology 2006, 7:17 Expression of human podocalyxin in HEK293 and Meg0l cells
treated with 5-AzaC
Figure 8
Expression of human podocalyxin in HEK293 and
Meg0l cells treated with 5-AzaC. Cells were treated
with 5-AzaC for 48 hrs. Cell lysates were resolved on SDS-
7.5% polyacrylamide gels under non-reducing conditions,
transferred to PVDF membranes and immunoblotted with A)
1 µg/ml of anti-podocalyxin mAb and B) 1 µg/ml of anti-Sp1
mAb. β-Actin was blotted as a loading control. The experi-
ment was repeated three times. degree of methylation of the promoter region and expres-
sion of podxl. The influence of the CpG methylation was
further assessed by treating the promoter constructs -111,-
181 and -210 with the methylase Sss I prior to transfection
into HEK293 cells. These results add further evidence
indicating that methylation may inhibit the activity of the
Podxl promoter. Methods
Materials As a
demethylating agent we used 5-azaC, structural analog of
citidine, that inactivates methyltransferases upon its
incorporation into DNA with the result of activation of
DNA transcription. This inhibitor has been used success-
fully as a therapeutic agent in acute myeloid leukemia and
myelodysplastic syndrome [32]. In our case, treatment of
low podxl-expressing cells with 5-azaC induced a dose-
dependent increase on podxl content whereas no effects
could be detected in cells, like HEK293, expressing higher
levels of podxl. However, the reported effect of 5-azaC in
inducing the activity of genes in the absence of methyla-
tion of CpG islands indicates that these results should be
taken cautiously [33]. The treatment with 5-azaC may sta-
bilize and increase the endogenous content of Sp1 [34]
nevertheless in our case 5-azaC did not modify Sp1 cellu-
lar content. On the other hand, bisulfite treatment of
genomic DNA from high (Tera-1) or low (Meg0l) podxl-
expressing cells showed a direct relationship between the Methods
Materials Cell culture media was from GIBCO, Invitrogen (Scot-
land, UK). Fetal bovine serum was from ICN Biochemi-
cals (Irvine, CA, USA). Restriction endonucleases were
obtained from Roche Diagnostics, SL (Barcelona, Spain). Antibody against β-actin was from Sigma RBI (Madrid,
Spain); mAbs against Sp-1 (sc-420x and sc-420) or AP2
(sc-25343x) were from Santa Cruz Biotechnology (Santa
Cruz, CA, USA). Mouse mAb against human podxl (clone
"3D3") was a generous gift of Dr. David B. Kershaw from
the Department of Pediatrics, University of Michigan. [α-
32P]dCTP was from Amersham Iberica (Spain). The Dual-
Luciferase Reporter Assay System, pGL3-basic vector and
Wizard DNA Clean-Up System were from Promega (Mad-
ison, WI). Sss I (CpG) methylase was from New England
Biolabs (Beverly, MA). Oligonucleotides were obtained
from Invitrogen (Paisley, UK). All other reagents were of
analytical grade and were purchased from commercial
sources. tion. We analyzed whether methylation of the Podxl pro-
moter could influence its transcriptional activity: firstly,
using demethylating agents in vivo and in second place by
studying the state of methylation of the CpG islands in
cultured cells expressing different amounts of Podxl. As a
demethylating agent we used 5-azaC, structural analog of
citidine, that inactivates methyltransferases upon its
incorporation into DNA with the result of activation of
DNA transcription. This inhibitor has been used success-
fully as a therapeutic agent in acute myeloid leukemia and
myelodysplastic syndrome [32]. In our case, treatment of
low podxl-expressing cells with 5-azaC induced a dose-
dependent increase on podxl content whereas no effects
could be detected in cells, like HEK293, expressing higher
levels of podxl. However, the reported effect of 5-azaC in
inducing the activity of genes in the absence of methyla-
tion of CpG islands indicates that these results should be
taken cautiously [33]. The treatment with 5-azaC may sta-
bilize and increase the endogenous content of Sp1 [34]
nevertheless in our case 5-azaC did not modify Sp1 cellu-
lar content. On the other hand, bisulfite treatment of
genomic DNA from high (Tera-1) or low (Meg0l) podxl-
expressing cells showed a direct relationship between the tion. We analyzed whether methylation of the Podxl pro-
moter could influence its transcriptional activity: firstly,
using demethylating agents in vivo and in second place by
studying the state of methylation of the CpG islands in
cultured cells expressing different amounts of Podxl. Discussion Transcriptional repression of methyl-
ated genes may be mediated by the binding of certain pro-
teins, such as MeCP2 [28-30]. Moreover, DNA
methylation may interfere with the binding of Sp1 to
DNA [31]. Demethylation of the promoter DNA may be a necessary
step in the epigenetic reprogramming of somatic cell
nuclei [19]. Demethylation is a selective process, operat-
ing only on the promoter, but not on enhancers; both a
putative Sp1/Sp3 and a GGGAGGG binding site are
required for demethylation and the initiation of transcrip- Page 6 of 12
(page number not for citation purposes) Page 6 of 12
(page number not for citation purposes) http://www.biomedcentral.com/1471-2199/7/17 BMC Molecular Biology 2006, 7:17 State of methylation of CpG dinucleotides in the human podocalyxin promoter
Figure 7
State of methylation of CpG dinucleotides in the human podocalyxin promoter. A) Schematic representation of a
fragment of the Podxl-pr comprising 208 nts upstream the transcription start site (indicated with an arrow) and the 5'UTR. Grey boxes denote the positions of CpG dinucleotides. Underlined nts indicate Sp1 recognition sites. B) Genomic DNA from
Tera-1 or Meg0l cells were extracted and subjected to bisulfite treatment, PCR amplification and DNA sequencing as
described in Methods. CpG sites are numbered from 1 to 27 relative to the transcription start site. Three bisulfite treatments
were performed. Data is expressed as % of methylation, calculated as percentage of conserved C in at least 10 clones obtained
after each bisulfite treatment. y
p
p
y
p
g
State of methylation of CpG dinucleotides in the human podocalyxin promoter. A) Schematic representation of a
fragment of the Podxl-pr comprising 208 nts upstream the transcription start site (indicated with an arrow) and the 5'UTR. Grey boxes denote the positions of CpG dinucleotides. Underlined nts indicate Sp1 recognition sites. B) Genomic DNA from
Tera-1 or Meg0l cells were extracted and subjected to bisulfite treatment, PCR amplification and DNA sequencing as
described in Methods. CpG sites are numbered from 1 to 27 relative to the transcription start site. Three bisulfite treatments
were performed. Data is expressed as % of methylation, calculated as percentage of conserved C in at least 10 clones obtained
after each bisulfite treatment. Page 7 of 12
(page number not for citation purposes) Page 7 of 12
(page number not for citation purposes) http://www.biomedcentral.com/1471-2199/7/17 Effect of in
Figure 9
Eff
f i ect o
v t o
et y at o o t e po oca y
p o ote
ve
uc e ase e p ess o
gu e 9
Effect of in vitro DNA methylation on the podocalyxin promoter-driven luciferase expression. Transient transfec-
tions were performed in HEK293 cells. Results represent mean ± SEM from three independent experiments performed in
duplicates. Promoter activity of each transfection was expressed as a ratio of luciferase/renilla activities. Data are expressed as
arbitrary units calculated as fold increases over void pGL-3 enhancer vector activity, which is assigned a value of 1.0. *p < 0.05
vs. mock methylated control. (Applied Biosystems, Foster City, CA). The TOPO con-
structs were digested with Sac I and Xho I and ligated to
the Sac I and Xho I sites of pGL3-basic plasmid. Restric-
tion enzyme digestion, sequence analysis, or both were
used to verify the constructions. A computer analysis for
potential binding sites on DNA sequences was carried out
with the TFSEARCH program [19]. Hind III sites of pGL3-basic to yield the construct -1297. Construct -1234 was produced by partial digestion with
EcoR I and religation of the -1297 construct. Constructs -
111, -181, -210, -364, -851, and -1108 were obtained by
PCR amplification from construct -1297 using as anti-
sense primer Podxl-pr 231/212 (5'-GCCTCGAGCAGGT-
GGCTGCGGT-3', underlined bp were mutated to generate
a Xho I site) and one of the following sense primers: Hind III sites of pGL3-basic to yield the construct -1297. Construct -1234 was produced by partial digestion with
EcoR I and religation of the -1297 construct. Constructs -
111, -181, -210, -364, -851, and -1108 were obtained by
PCR amplification from construct -1297 using as anti-
sense primer Podxl-pr 231/212 (5'-GCCTCGAGCAGGT-
GGCTGCGGT-3', underlined bp were mutated to generate
a Xho I site) and one of the following sense primers: Cell lines Podxl-pr -111/-93 (5'-TGGCGAGCTCCTGGGGGAGGG-
3'),
Podxl-pr
-181/-161
(5'-GCTCAGAGCTCACG-
GCCCAGCG-3'), Podxl-pr -210/-189 (5'-GTCGAGCTC-
CAGGGCGGAGTTTC-3'),
Podxl-pr
-364/343
(5'-
GGGAGCTCGCCGGGCCCACTTA-3'), Podxl-pr -851/-
830 (5'-CTGAGCTCAGAGGCAGGTTTGC-3'), Podxl-pr -
1108/-1087
(5'-TTGAGCTCCCCAACAACCCCCT-3'). Underlined nucleotides in sense oligonucleotides were
mutated to generate Sac I sites. pGL3-constructs with
mutated Sp1 sites were obtained by PCR from DNA tem-
plate and antisense oligonucleotide mentioned above and
the following sense primers: Podxl-pr -90/-79 mutSp1-A
(5'-GAGCTCCGGTGGTGGGGGTGGGGG-3'), Podxl-pr -
146/-126
mutSp1-B
(5'-CAGCTCCGGCCTTGCCCG-
GCC-3'), Podxl-pr -178/-150 mutSp1-C (5'-TCATTGT-
TCAAAGCCCAGAGCCCACCCAG-3'), Podxl-pr -207/-
184 mutSp1-C (5'-GAGCTCCAGGTTGGAGTTTCC-3'). Underlined nucleotides show mutated nucleotides. The
resulting PCR products were subcloned into a T vector and
sequenced in an ABI PRISM 3100 Genetic Analyzer HepG2 (hepatoma cells) and HEK293 (human embry-
onic kidney cells) were grown in DMEM, Tera-1 (testes
embryonal carcinoma cells) in McCoy medium, Meg0l
(human megakaryoblastic cells) in RPMI medium, all
obtained from GIBCO (Madrid, Spain), and Drosophila
Schneider's SL2 cells in Schneider's insect medium
(Sigma, Madrid, Spain). Culture media were supple-
mented with 10% fetal bovine serum. SL2 cells were
grown at 25°C and all other cell lines at 37°C. Construction of reporter plasmids p
p
A 1528-bp DNA fragment was amplified by PCR from
human genomic DNA using the primers Podxl-pr -1297/-
1275
(5'-GAGCGGGGGAGGGGAAAGGAA-3')
and
Podxl-pr 231/208 (5'-GGAGCAGGTGGCTGCGGTGC-3')
as sense and antisense oligonucleotides, respectively. The
PCR product was subcloned directly into a T vector (Topo
TA cloning, Invitrogen) to introduce Xho I and Hind III
restriction sites in 5' and 3' ends, respectively. The Xho I/
Hind III (-1297/231) fragment was subcloned into Xho I/ Page 8 of 12
(page number not for citation purposes) BMC Molecular Biology 2006, 7:17 http://www.biomedcentral.com/1471-2199/7/17 Effect of in vitro DNA methylation on the podocalyxin promoter-driven luciferase expression
Figure 9
Effect of in vitro DNA methylation on the podocalyxin promoter-driven luciferase expression. Transient transfec-
tions were performed in HEK293 cells. Results represent mean ± SEM from three independent experiments performed in
duplicates. Promoter activity of each transfection was expressed as a ratio of luciferase/renilla activities. Data are expressed as
arbitrary units calculated as fold increases over void pGL-3 enhancer vector activity, which is assigned a value of 1.0. *p < 0.05
vs. mock methylated control. Page 9 of 12
(page number not for citation purposes) Gel retardation assay DNA probes were prepared by PCR amplification from
the -1296 construct. The following sense and antisense
primers were used: for -243/-18 probe: Podxl-pr -243/-
223 (5'-CCAAGGCTCGAGGCCGCCGG-3') and Podxl-pr
-18/-38 (5'-CACTGGCGGCTGCGGCTACTC-3'); for-538/
-238 probe: Podxl-pr-538/-518 (5'-GGGCTCGAGAG-
GCAGGTGAAT-3') and Podxl-pr -238/-258 (5'-CCTT-
GGGGGGCTGTGTGTGCG-3'), for A probe: Podxl-pr -
111/-93 and Podxl-pr -18/-38, for mutSp1-A probe:
Podxl-pr -90/-79 mutSp1-A and Podxl-pr -18/-38, for B
probe:
Podxl-pr
-146/-136
(5'GAGCTCCG-
GCCCCGCCCGGCC-3') and Podxl-pr -94/-114 (5'CTC-
GAGCAGGAGCCCGCCAGC-3'); for mutSp1-B probe:
Podxl-pr-146/-126 mutSp1-B and Podxl-pr -94/-114; for
C probe: Podxl-pr-181/161 and Podxl-pr -94/-114; for
mutSp1-C probe: hPodxl-pr -178/-150 mutSp1-C and
Podxl-pr -94/-114; for D probe: Podxl-pr -210/-189 and
Podxl-pr
-150/-178mutSp1-C
(5'-CTGGGTTGGTTCT-
GGGCTTTGAATGA-3'); for mutSp1-D probe: Podxl-pr -
207/-184 mutSp1-D and Podxl-pr -150/-178mutSp1-C. All the probes were labeled using the Klenow fragment of
Escherichia coli polymerase and [32P]dCTP. Nuclear
extracts from Tera-1 and Meg0l cells were prepared
according to standard protocols. Nuclear extracts (5 µg)
were preincubated for 15 min at 4°C in 25 µL-binding
reaction containing: 10 mM Tris/HCl pH 7.5, 50 mM
NaCl, 2.5 mM MgC12, 4% glycerol, 0.5 mM dithiothreitol
and 3 µg poly(dI-dC). After adding ~30 pg of labeled
DNA, the mixture was incubated for 30 min at room tem-
perature, and the complexes formed resolved on a 4%
nondenaturing polyacrylamide gel in 0.5 × TBE buffer. Gels were run at 150 V at room temperature, dried and
exposed at -80°C to an X-ray film. Transient transfection procedure HepG2, Tera-1 and HEK293 cells were transfected by the
calcium phosphate precipitation technique in 60-mm
dishes with 2 µg of pGL3/Podxl constructs described
above and 0.5 µg of phRL-cmv plasmid for transfection
efficiency correction. For negative and positive controls of
transfection, pGL3-basic and the 3TP promoter-luciferase
construct (p3TP-lux), containing three copies of the TPA
response element from the human collagenase promoter
plasmids were used. SL2 cells were cotransfected with Page 9 of 12
(page number not for citation purposes) Page 9 of 12
(page number not for citation purposes) http://www.biomedcentral.com/1471-2199/7/17 BMC Molecular Biology 2006, 7:17 pGL3/Podxl constructs and either pPAC-Sp1 that
expresses human Sp1 driven by the Drosophila actin pro-
moter, or the control vector pPacO, containing only the
Drosophila actin promoter. The amount of transfected
DNA was maintained constant with void plasmid. Either
6 h (HEK293 and Tera-1 cells) or 16 h (HepG2 and SL2
cells) after transfection, the medium was changed and
cells were incubated for another 48 h. The luciferase activ-
ity was determined using the Dual-Luciferase Reporter
Assay System (Promega, Madison, WI, USA) in cell
extracts prepared according to manufacturer recommen-
dations in a TD-20/20 luminometer (Turner Designs,
Sunnyvale, CA, USA). Luciferase activities were normal-
ized by Renilla luciferase activity, except for SL2 experi-
ments that were normalized by the protein concentration
that was measured using a Bradford protein assay kit (Bio-
Rad Laboratories, Inc., Richmond, CA, USA). ing sites for AP2, Sp1, NFkB and GATA transcription
factors were included in the binding reaction. These DNA
fragments were prepared by hybridization of sense and
antisense oligonucleotides of the following sequences: 5'-
GATCGAACTGACCGCCCGCGGCCC-3' for AP2, 5'-
ATTCGATCGGGGCGGGGCGAG-3' for Sp1, 5'-AGTT-
GAGGGGACTTTCCCAGGC-3' for NFkB and 5'-CACTT-
GATAACAGAAAGTGATAACTCT-3'
for
GATA
DNA
binding sites. As nonspecific competitor, an unrelated
DNA fragment of similar size and base composition was
used. In supershift experiments, nuclear extracts were pre-
incubated with either mAbs against Sp1, AP2 or nonspe-
cific antibodies, prior to incubation with the radiolabeled
probes. Treatment of cells with 5-Aza-2 deoxycytidine The cells were treated with 5-azaC at a final concentration
of 1 µM or 5 µM for 7 days. The medium was changed
every second day. Negative controls were treated in the
same manner without 5-azaC. Then, cells were lysed for
western blot analysis and blotted with the mAb anti-Podxl
primary antibody (3D3). In Vitro methylation reactions To examine the effect of CG methylation on the promoter
activity, Podxl promoter constructs -111,-181,-210 were
mock methylated or methylated by Sss I methylase in the
absence or presence of S-adenosylmethionine. The status
of methylation was determined by digestion with the
restriction enzyme Hpa II. Mock-methylated or Sss I
methylated plasmids were isolated from 1% agarose gel
using Gel Purification Kit from Qiagen. Western blot analysis For western blot analysis, cells were lysed in 100 µl of
phosphate buffered saline containing 1% Triton X-100,
0.1% SDS, 2 mM phenylmethylsulfonyl fluoride, 2 mM
EDTA, and 1% of protease inhibitors cocktail. Cellular
extracts were resolved by SDS-PAGE and transferred to
PVDF membranes. The membranes were incubated with
mAb against Sp1 (Sta Cruz sc-420) or Podxl (3D3) and
subsequently with the horseradish peroxidase-conjugated
secondary antibody (BioRad, Madrid, Spain). β-Actin was
blotted in order to normalize the amount of lysate in the
experiments. To develop the blots we use the ECL reagent. References 23. Casey G, Neville PJ, Liu X, Plummer SJ, Cicek MS, Krumroy LM, Cur-
ran AP, McGreevy MR, Catalona WJ, Klein EA, Witte JS: Podoca-
lyxin Variants and Risk of Prostate Cancer and Tumor
Aggressiveness. Hum Mol Genet in press. 2006, Jan 24 1. Kerjaschki D, Sharkey DJ, Farquhar MG: Identification and char-
acterization of podocalyxin-the major sialoprotein of the
renal glomerular epithelial cell. J Cell Biol 1984, 98:1591-1596. 1. Kerjaschki D, Sharkey DJ, Farquhar MG: Identification and char-
acterization of podocalyxin-the major sialoprotein of the
renal glomerular epithelial cell. J Cell Biol 1984, 98:1591-1596. g
p
J
,
2. Dekan G, Gabel C, Farquhar MG: Sulfate contributes to the neg-
ative charge of podocalyxin, the major sialoglycoprotein of
the glomerular filtration slits. Proc Natl Acad Sci USA 1991,
88:5398-5402. g
p
J
2. Dekan G, Gabel C, Farquhar MG: Sulfate contributes to the neg-
ative charge of podocalyxin, the major sialoglycoprotein of
the glomerular filtration slits. Proc Natl Acad Sci USA 1991,
88:5398-5402. gg
p
J
24. Kelley TW, Huntsman D, McNagny KM, Roskelly CD, His ED: Podo-
calyxin: A marker of blasts in acute leukemia. Am J Clin Pathol
2005, 124:134-142. 25. Kreidberg JA, Sariola H, Loring JM, Maeda M, Pelletier J, Housman D,
Jaenisch R: WT-1 is required for early kidney development. Cell 1993, 74:679-691. 3. Seiler MW, Rennke HG, Venkatachalam MA, Cotran RS: Pathogen-
esis of polycation-induced alterations ("fusion") of glomeru-
lar epithelium. Lab Invest 1977, 36:48-61. 26. Moore AW, McInnes L, Kreidberg J, Hastie ND, Schedl A: YAC
complementation shows a requirement for Wt1 in the
development of epicardium, adrenal gland and throughout
nephrogenesis. Development 1999, 126:1845-1857. p
4. Andrews PM: Glomerular epithelial alterations resulting from
sialic acid surface coat removal. Kidney Int 1979, 15:376-385. 4. Andrews PM: Glomerular epithelial alterations resulting from
sialic acid surface coat removal. Kidney Int 1979, 15:376-385. y
5. Doyonnas , Kershaw DB, Duhme C, Merkens H, Chelliah S, Graf T,
McNagny KM: Anuria, omphalocele, and perinatal lethality in
mice lacking the CD34-related protein podocalyxin. J Exp
Med 2001, 194:13-27. 5. Doyonnas , Kershaw DB, Duhme C, Merkens H, Chelliah S, Graf T,
McNagny KM: Anuria, omphalocele, and perinatal lethality in
mice lacking the CD34-related protein podocalyxin. J Exp
Med 2001, 194:13-27. 27. Robertson KD: DNA methylation and human disease. Nat Rev
Genet 2005, 6:597-610. 28. Competing interests The author(s) declare they have no competing interests. 11. Fina L, Molgaard HV, Robertson D, Bradley NJ, Monaghan P, Delia D,
Sutherland DR, Baker MA, Greaves MF: Expression of the CD34
gene in vascular endothelial cells. Blood 1990, 75:2417-2426. Bisulfite modification of genomic DNA To release small fragments of genomic DNA of Meg0l and
Tera-1 cells we digested with restriction enzymes that
don't cleave the CpG region of the Podxl-pr. Digested
genomic DNAs were denatured, modified by bisulfite,
purified, and desulphonated according to Herman et al
[35]. Degenerated primers for PCR were designed to
amplify the region between -618 to +368 (Podxl-pr -618/
-592 5'-GGTGAAGGAGTGAATTAGTC/TC/TAGC/TTTG-3' For competition assays, 100-fold molar excess of unla-
beled double stranded DNA consensus sequences of bind- Page 10 of 12
(page number not for citation purposes) Page 10 of 12
(page number not for citation purposes) http://www.biomedcentral.com/1471-2199/7/17 http://www.biomedcentral.com/1471-2199/7/17 BMC Molecular Biology 2006, 7:17 and Podxl-pr 368/343 5'-AGAATGGTGAGTGGC/TC/
TGGC/TC/TTGGGTT-3). The PCR fragments were purified
by the JetQuick Purification Kit (Genomed) and
sequenced. and Podxl-pr 368/343 5'-AGAATGGTGAGTGGC/TC/
TGGC/TC/TTGGGTT-3). The PCR fragments were purified
by the JetQuick Purification Kit (Genomed) and
sequenced. 9. Hara T, Nakano Y, Tanaka M, Tamura K, Sekiguchi T, Minehata K,
Copeland NG, Jenkins NA, Okabe M, Kogo H, Mukouyama Y, Miya-
jima A: Identification of podocalyxin-like protein 1 as a novel
cell surface marker for hemangioblasts in the murine aorta-
gonad-mesonephros region. Immunity 1999, 11:567-578. 10. Kerjaschki D, Poczewski H, Dekan G, Horvat R, Balzar E, Kraft N,
Atkins RC: Identification of a major sialoprotein in the glyco-
calyx of human visceral glomerular epithelial cells. J Clin Invest
1986, 78:1142-1149. Authors' contributions 12. Sassetti C, Tangemann K, Singer MS, Kershaw DS, Rosen SD: Identi-
fication of podocalyxin-like protein as a high endothelial
venule ligand for L-selectin: parallels to CD34. J Exptl Med
1998, 187:1965-1975. NB participated in the experimental design, helped to
draft the manuscript and carried out experiments shown
in Figs. 2 to 6 and collaborated in experiments shown in
Figs 1 and 9. SL performed experiments shown in Fig. 5
and contributed to the conceptual basis of the study. SA,
RBR and CGM performed the computer analysis of the
promoter region and prepared some of the constructs
used along this work. EGA-S and MSA participated in the
experiments shown in Figs. 3 and 4. RP designed, coordi-
nated and drafted the manuscript. All authors approved
the manuscript. 13. Sassetti C, Van Zante A, Rose SD: Identification of endoglycan, a
member of the CD34/Podocalyxin family of sialomucins. J
Biol Chem 2000, 275:9001-9010. 14. Palmer RE, Kotsianti A, Cadman B, Boyd T, Gerald W, Haber DA:
WT1 regulates the expression of the major glomerular
podocyte membrane protein Podocalyxin. Current Biology
2001, 11:1805-1809. 15. Kershaw DB, Wiggins JE, Wharram BL, Wiggins RC: Assignment of
the human podocalyxin-like protein (PODXL) gene to 7q32-
q33. Genomics 1997, 45:239-40. 16. Akiyama Y: TFSEARCH: Searching Transcription Factor
Binding Sites. [http://www.cbrc.jp/research/db/TFSEARCH.html]. 17. Siegfried Z, Eden S, Mendelsohn M, Feng X, Tsuberi BZ, Cedar H:
DNA methylation represses transcription in vivo. Nat Genet
1999, 22:203-206. Acknowledgements g
g
DNA methylation represses transcription in vivo. Nat Genet
1999, 22:203-206. N. Butta is recipient of a tenure track grant Ramon y Cajal from the Spanish
Ministry of Science. S. Larrucea was supported by a contract from the
Comunidad de Madrid (08.4/0015.1/2001) and at present holds a research
tenure track of the Spanish Council of Research. S. Alonso was recipient of
a predoctoral fellowship from the Basque Autonomous Community
(BF101-40). E G Arias-Salgado is recipient of a tenure research grant Juan
de la Cierva. This work has been supported in part by grants from the
Direccion General de Investigacion (SAP 2004-04345), Fondo de Investiga-
ciones Sanitarias (FIS-PI021263 and PI050546) and Comunidad de Madrid
(08.4/0029.1/2003). N. Butta was recipient of a grant from the Fundacion
Rodríguez Pascual. We thank Dr. Kershaw for the generous gift of the mAb
3D3. 18. Hattori N, Nishino K, Ko YG, Hattori N, Ohgane J, Tanaka S, Shiota
K: Epigenetic control of mouse Oct-4 gene expression in ES
cells and TS cells. J Biol Chem 2004, 279:17063-17069. J
19. Simonsson S, Gurdon J: DNA demethylation is necessary for the
epigenetic reprogramming of somatic cell nuclei. Nat Cell Biol
2004, 6:984-989. 20. European Bioinformatic Institute: CpGPlot program. [http://
www.ebi.ac.uk/emboss/cpgplot/]. pgp
]
21. Schopperle WM, Kershaw DB, DeWolf WC: Human embryonal
carcinoma tumor antigen, Gp200/GCTM-2 is podocalyxin. Biochem Biophys Res Commun 2003, 399:285-290. p y
22. Somasiri A, Nielsen JS, Makretsov N, McCoy ML, Prentice L, Gilks CB,
Chia SK, Gelmon KA, Kershaw DB, Huntsman DG, McNagny KM,
Roskelley CD: Overexpression of anti-adhesin podocalyxin is
an independent predictor of breast cancer progression. Can-
cer Res 2004, 64:5068-5073. References Jones PL, Veenstra GJ, Wade PA, Vermaak D, Kass SU, Landsberger
N, Strouboulis J, Wolffe AP: Methylated DNA and MeCP2
recruit histone deacetylase to repress transcription. Nat
Genet 1998, 19:187-191. 6. Kershaw DB, Thomas PE, Wharram BL, Goyal M, Wiggins JE, White-
side CI, Wiggins RC: Molecular cloning, expression and charac-
terization of podocalyxin-like protein from rabbit as a
transmembrane protein of glomerular podocytes and vascu-
lar endothelium. J Biol Chem 1995, 270:29439-29446. 29. Nan X, Ng HH, Johnson CA, Laherty CD, Turner BM, Eisenman RN,
Bird A: Transcriptional repression by the methyl-CpG-bind-
ing protein MeCP2 involves a histone deacetylase complex. Nature 1998, 393:386-389. J
7. Kershaw DB, Beck SG, Wharram BL, Wiggins JE, Goyal M, Thomas
PE, Wiggins RC: Molecular cloning and characterization of
human podocalyxin-like protein. J Biol Chem 1997,
272:15708-15714. 30. Bird AP, Wolffe AP: Methylation-induced repression-belts,
braces, and chromatin. Cell 1999, 99:451-454. 8. McNagny KM, Petterson I, Rossi F, Flamme A, Shevchenko A, Mann
M, Graf T: Thrombomucin, a novel cell surface protein that
defines thrombocytes and multipotent hematopoietic pro-
genitors. J Cell Biol 1997, 138:1395-1407. 31. Cao YX, Jean JC, Williams MC: Cytosine methylation of an Sp1
site contributes to organ-specific and specific regulation of Page 11 of 12
(page number not for citation purposes) Page 11 of 12
(page number not for citation purposes) BMC Molecular Biology 2006, 7:17 http://www.biomedcentral.com/1471-2199/7/17 http://www.biomedcentral.com/1471-2199/7/17 http://www.biomedcentral.com/1471-2199/7/17 expression of the lung epithelial gene T1. Biochem J 2000,
350:883-890. 32. Daskalakis M, Nguyen TT, Nguyen C, Guldberg P, Köhler G, Wijer-
mans P, Jones PA, Lubbert M: Demethylation of a hypermethyl-
ated p16/INK4B gene in patients with myeloblastic
syndrome by 5-Aza-2'-dexycytidine (decitabine) treatment. Blood 2002, 100:2957-2964. 33. Schmelz K, Sattler N, Wagner M, Lubbert M, Dorken B, Tamm I:
Induction of gene expression by 5-Aza-2'-deoxycytidine in
acute myeloid leukemia (AML) and myelodysplastic syn-
drome (MDS) but not epithelial cells by DNA-methylation-
dependent and -independent mechanisms. Leukemia 2004,
28:1-9. 34. Ammanamanchi S, Kim SJ, Sun LZ, Brattain MG: Induction of trans-
forming growth factor-beta receptor type II expression in
estrogen receptor-positive breast cancer cells through SP1
activation by 5-aza-2'-deoxycytidine. J Biol Chem 1998,
273:16527-16534. 35. Herman JG, Graff jr, Myöhänen S, Nelkin BD, Baylin SB: Methyla-
tion-specific PCR: A novel PCR assay for methylation status
of CpG islands. Proc Natl Acad Sci USA 1996, 93:9821-9826. References Publish with BioMed Central and every
scientist can read your work free of charge
"BioMed Central will be the most significant development for
disseminating the results of biomedical research in our lifetime."
Sir Paul Nurse, Cancer Research UK
Your research papers will be:
available free of charge to the entire biomedical community
peer reviewed and published immediately upon acceptance
cited in PubMed and archived on PubMed Central
yours — you keep the copyright
Submit your manuscript here:
http://www.biomedcentral.com/info/publishing_adv.asp
BioMedcentral
Page 12 of 12
(page number not for citation purposes) Publish with BioMed Central and every
scientist can read your work free of charge Page 12 of 12
(page number not for citation purposes)
|
Subsets and Splits
No community queries yet
The top public SQL queries from the community will appear here once available.